Sample records for unraveling neuronal thy-1
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Maldonado, H; Calderon, C; Burgos-Bravo, F; Kobler, O; Zuschratter, W; Ramirez, O; Härtel, S; Schneider, P; Quest, A F G; Herrera-Molina, R; Leyton, L
2017-02-01
Two key proteins for cellular communication between astrocytes and neurons are αvβ3 integrin and the receptor Thy-1. Binding of these molecules in the same (cis) or on adjacent (trans) cellular membranes induces Thy-1 clustering, triggering actin cytoskeleton remodeling. Molecular events that could explain how the Thy-1-αvβ3 integrin interaction signals have only been studied separately in different cell types, and the detailed transcellular communication and signal transduction pathways involved in neuronal cytoskeleton remodeling remain unresolved. Using biochemical and genetic approaches, single-molecule tracking, and high-resolution nanoscopy, we provide evidence that upon binding to αvβ3 integrin, Thy-1 mobility decreased while Thy-1 nanocluster size increased. This occurred concomitantly with inactivation and exclusion of the non-receptor tyrosine kinase Src from the Thy-1/C-terminal Src kinase (Csk)-binding protein (CBP)/Csk complex. The Src inactivation decreased the p190Rho GTPase activating protein phosphorylation, promoting RhoA activation, cofilin, and myosin light chain II phosphorylation and, consequently, neurite shortening. Finally, silencing the adaptor CBP demonstrated that this protein was a key transducer in the Thy-1 signaling cascade. In conclusion, these data support the hypothesis that the Thy-1-CBP-Csk-Src-RhoA-ROCK axis transmitted signals from astrocytic integrin-engaged Thy-1 (trans) to the neuronal actin cytoskeleton. Importantly, the β3 integrin in neurons (cis) was not found to be crucial for neurite shortening. This is the first study to detail the signaling pathway triggered by αvβ3, the endogenous Thy-1 ligand, highlighting the role of membrane-bound integrins as trans acting ligands in astrocyte-neuron communication. Copyright © 2016 Elsevier B.V. All rights reserved.
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Taylor-Clark, Thomas E.; Wu, Kevin Y.; Thompson, Julie-Ann; Yang, Kiseok; Bahia, Parmvir K.; Ajmo, Joanne M.
2015-01-01
The Thy1.2 YFP-16 mouse expresses yellow fluorescent protein (YFP) in specific subsets of peripheral and central neurons. The original characterization of this model suggested that YFP was expressed in all sensory neurons, and this model has been subsequently used to study sensory nerve structure and function. Here, we have characterized the expression of YFP in the sensory ganglia (DRG, trigeminal and vagal) of the Thy1.2 YFP-16 mouse, using biochemical, functional and anatomical analyses. Despite previous reports, we found that YFP was only expressed in approximately half of DRG and trigeminal neurons and less than 10% of vagal neurons. YFP-expression was only found in medium and large-diameter neurons that expressed neurofilament but not TRPV1. YFP-expressing neurons failed to respond to selective agonists for TRPV1, P2X2/3 and TRPM8 channels in Ca2+ imaging assays. Confocal analysis of glabrous skin, hairy skin of the back and ear and skeletal muscle indicated that YFP was expressed in some peripheral terminals with structures consistent with their presumed non-nociceptive nature. In summary, the Thy1.2 YFP-16 mouse expresses robust YFP expression in only a subset of sensory neurons. But this mouse model is not suitable for the study of nociceptive nerves or the function of such nerves in pain and neuropathies. PMID:25746468
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Laperchia, Claudia; Allegra Mascaro, Anna L.; Sacconi, Leonardo; Andrioli, Anna; Mattè, Alessandro; De Franceschi, Lucia; Grassi-Zucconi, Gigliola; Bentivoglio, Marina; Buffelli, Mario; Pavone, Francesco S.
2013-01-01
Transgenic mice expressing fluorescent proteins in specific cell populations are widely used for in vivo brain studies with two-photon fluorescence (TPF) microscopy. Mice of the thy1GFP-M line have been engineered for selective expression of green fluorescent protein (GFP) in neuronal populations. Here, we report that TPF microscopy reveals, at the brain surface of these mice, also motile non-neuronal GFP+ cells. We have analyzed the behavior of these cells in vivo and characterized in brain sections their immunophenotype. With TPF imaging, motile GFP+ cells were found in the meninges, subarachnoid space and upper cortical layers. The striking feature of these cells was their ability to move across the brain parenchyma, exhibiting evident shape changes during their scanning-like motion. In brain sections, GFP+ cells were immunonegative to antigens recognizing motile cells such as migratory neuroblasts, neuronal and glial precursors, mast cells, and fibroblasts. GFP+ non-neuronal cells exhibited instead the characteristic features and immunophenotype (CD11c and major histocompatibility complex molecule class II immunopositivity) of dendritic cells (DCs), and were immunonegative to the microglial marker Iba-1. GFP+ cells were also identified in lymph nodes and blood of thy1GFP-M mice, supporting their identity as DCs. Thus, TPF microscopy has here allowed the visualization for the first time of the motile behavior of brain DCs in situ. The results indicate that the thy1GFP-M mouse line provides a novel animal model for the study of subsets of these professional antigen-presenting cells in the brain. Information on brain DCs is still very limited and imaging in thy1GFP-M mice has a great potential for analyses of DC-neuron interaction in normal and pathological conditions. PMID:23409142
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SUZUKI, Maasa; EBARA, Satomi; KOIKE, Taro; TONOMURA, Sotatsu; KUMAMOTO, Kenzo
2012-01-01
Hairs are known as a sensory apparatus for touch. Their follicles are innervated predominantly by palisade endings composed of longitudinal and circumferential lanceolate endings. However, little is known as to how their original primary neurons make up a part of the ending. In this study, innervation of the palisade endings was investigated in the auricular skin of thy1-YFP transgenic mouse. Major observations were 1) Only a small portion of PGP9.5-immunopositive axons showed YFP-positivity, 2) All of thy1-YFP-positive sensory axons were thick and myelinated, 3) Individual thy1-YFP-positive trunk axons innervated 4–54 hair follicles, 4) Most palisade endings had a gap of lanceolate ending arrangement, 5) PGP9.5-immunopositive 10–32 longitudinal lanceolate endings were closely arranged. Only a part of them were thy1-YFP-positive axons that originated from 1–3 afferents, and 6) Single nerve bundles of the dermal nerve network included both bidirectional afferents. Palisade endings innervated by multiple sensory neurons might be highly sensitive to hair movement. PMID:23229751
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Suzuki, Maasa; Ebara, Satomi; Koike, Taro; Tonomura, Sotatsu; Kumamoto, Kenzo
2012-01-01
Hairs are known as a sensory apparatus for touch. Their follicles are innervated predominantly by palisade endings composed of longitudinal and circumferential lanceolate endings. However, little is known as to how their original primary neurons make up a part of the ending. In this study, innervation of the palisade endings was investigated in the auricular skin of thy1-YFP transgenic mouse. Major observations were 1) Only a small portion of PGP9.5-immunopositive axons showed YFP-positivity, 2) All of thy1-YFP-positive sensory axons were thick and myelinated, 3) Individual thy1-YFP-positive trunk axons innervated 4-54 hair follicles, 4) Most palisade endings had a gap of lanceolate ending arrangement, 5) PGP9.5-immunopositive 10-32 longitudinal lanceolate endings were closely arranged. Only a part of them were thy1-YFP-positive axons that originated from 1-3 afferents, and 6) Single nerve bundles of the dermal nerve network included both bidirectional afferents. Palisade endings innervated by multiple sensory neurons might be highly sensitive to hair movement.
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Imaging Neural Activity Using Thy1-GCaMP Transgenic mice
Chen, Qian; Cichon, Joseph; Wang, Wenting; Qiu, Li; Lee, Seok-Jin R.; Campbell, Nolan R.; DeStefino, Nicholas; Goard, Michael J.; Fu, Zhanyan; Yasuda, Ryohei; Looger, Loren L.; Arenkiel, Benjamin R.; Gan, Wen-Biao; Feng, Guoping
2014-01-01
Summary The ability to chronically monitor neuronal activity in the living brain is essential for understanding the organization and function of the nervous system. The genetically encoded green fluorescent protein based calcium sensor GCaMP provides a powerful tool for detecting calcium transients in neuronal somata, processes, and synapses that are triggered by neuronal activities. Here we report the generation and characterization of transgenic mice that express improved GCaMPs in various neuronal subpopulations under the control of the Thy1 promoter. In vitro and in vivo studies show that calcium transients induced by spontaneous and stimulus-evoked neuronal activities can be readily detected at the level of individual cells and synapses in acute brain slices, as well as chronically in awake behaving animals. These GCaMP transgenic mice allow investigation of activity patterns in defined neuronal populations in the living brain, and will greatly facilitate dissecting complex structural and functional relationships of neural networks. PMID:23083733
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Thy-1, the enigmatic extrovert on the neuronal surface.
Morris, R
1992-10-01
Thy-1 is a small glycoprotein of 110 amino acids which, folded in the characteristic structure of an immunoglobulin variable domain, are enchored to the plasma membrane via a glycophosphatidylinositol (GPI) tail (Fig. 1). It is a major component of the surface of various cell types, including neurons, at certain stages of their development. These qualities doubtlessly appeal to certain cognoscenti, but it is not clear why they would raise Thy-1 to the status of a favourite molecule. Indeed, few scientists readily admit to having a favourite. We study individual molecules because science is rooted in specific observations; but we do so in order to discover mechanisms of general importance. A molecule's appeal is dependent on its ability to reveal novel aspects of how nature works. Thy-1 has been unusual in this respect. It was the first lymphocyte surface antigen shown to be restricted to a functional subset of lymphocytes (T cells in the mouse), a finding crucial to the development of cellular immunology; it was one of the first cell surface molecules to be sequenced and indicated the importance of immunoglobulin domains and GPI anchors as structural motifs; it has been pivotal in studies demonstrating that GPI-anchored molecules are able to signal across the membrane they do not span. Thy-1 has revealed this much, however, with the charm of an adroit stripper: it has always promised glimpses of things more exciting than that displayed. In particular, the function of this molecule has never emerged.(ABSTRACT TRUNCATED AT 250 WORDS)
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Bu, Wei; Ren, Huiling; Deng, Yunping; Del Mar, Nobel; Guley, Natalie M.; Moore, Bob M.; Honig, Marcia G.; Reiner, Anton
2016-01-01
We have previously reported that mild TBI created by focal left-side cranial blast in mice produces widespread axonal injury, microglial activation, and a variety of functional deficits. We have also shown that these functional deficits are reduced by targeting microglia through their cannabinoid type-2 (CB2) receptors using 2-week daily administration of the CB2 inverse agonist SMM-189. CB2 inverse agonists stabilize the G-protein coupled CB2 receptor in an inactive conformation, leading to increased phosphorylation and nuclear translocation of the cAMP response element binding protein (CREB), and thus bias activated microglia from a pro-inflammatory M1 to a pro-healing M2 state. In the present study, we showed that SMM-189 boosts nuclear pCREB levels in microglia in several brain regions by 3 days after TBI, by using pCREB/CD68 double immunofluorescent labeling. Next, to better understand the basis of motor deficits and increased fearfulness after TBI, we used unbiased stereological methods to characterize neuronal loss in cortex, striatum, and basolateral amygdala (BLA) and assessed how neuronal loss was affected by SMM-189 treatment. Our stereological neuron counts revealed a 20% reduction in cortical and 30% reduction in striatal neurons bilaterally at 2–3 months post blast, with SMM-189 yielding about 50% rescue. Loss of BLA neurons was restricted to the blast side, with 33% of Thy1+ fear-suppressing pyramidal neurons and 47% of fear-suppressing parvalbuminergic (PARV) interneurons lost, and Thy1-negative fear-promoting pyramidal neurons not significantly affected. SMM-189 yielded 50–60% rescue of Thy1+ and PARV neuron loss in BLA. Thus, fearfulness after mild TBI may result from the loss of fear-suppressing neuron types in BLA, and SMM-189 may reduce fearfulness by their rescue. Overall, our findings indicate that SMM-189 rescues damaged neurons and thereby alleviates functional deficits resulting from TBI, apparently by selectively modulating microglia to the beneficial M2 state. CB2 inverse agonists thus represent a promising therapeutic approach for mitigating neuroinflammation and neurodegeneration. PMID:27766068
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Lipid Rafts Act as Specialized Domains for Tetanus Toxin Binding and Internalization into Neurons
Herreros, Judit; Ng, Tony; Schiavo, Giampietro
2001-01-01
Tetanus (TeNT) is a zinc protease that blocks neurotransmission by cleaving the synaptic protein vesicle-associated membrane protein/synaptobrevin. Although its intracellular catalytic activity is well established, the mechanism by which this neurotoxin interacts with the neuronal surface is not known. In this study, we characterize p15s, the first plasma membrane TeNT binding proteins and we show that they are glycosylphosphatidylinositol-anchored glycoproteins in nerve growth factor (NGF)-differentiated PC12 cells, spinal cord cells, and purified motor neurons. We identify p15 as neuronal Thy-1 in NGF-differentiated PC12 cells. Fluorescence lifetime imaging microscopy measurements confirm the close association of the binding domain of TeNT and Thy-1 at the plasma membrane. We find that TeNT is recruited to detergent-insoluble lipid microdomains on the surface of neuronal cells. Finally, we show that cholesterol depletion affects a raft subpool and blocks the internalization and intracellular activity of the toxin. Our results indicate that TeNT interacts with target cells by binding to lipid rafts and that cholesterol is required for TeNT internalization and/or trafficking in neurons. PMID:11598183
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Neuronal and astrocytic metabolism in a transgenic rat model of Alzheimer's disease.
Nilsen, Linn Hege; Witter, Menno P; Sonnewald, Ursula
2014-05-01
Regional hypometabolism of glucose in the brain is a hallmark of Alzheimer's disease (AD). However, little is known about the specific alterations of neuronal and astrocytic metabolism involved in homeostasis of glutamate and GABA in AD. Here, we investigated the effects of amyloid β (Aβ) pathology on neuronal and astrocytic metabolism and glial-neuronal interactions in amino acid neurotransmitter homeostasis in the transgenic McGill-R-Thy1-APP rat model of AD compared with healthy controls at age 15 months. Rats were injected with [1-(13)C]glucose and [1,2-(13)C]acetate, and extracts of the hippocampal formation as well as several cortical regions were analyzed using (1)H- and (13)C nuclear magnetic resonance spectroscopy and high-performance liquid chromatography. Reduced tricarboxylic acid cycle turnover was evident for glutamatergic and GABAergic neurons in hippocampal formation and frontal cortex, and for astrocytes in frontal cortex. Pyruvate carboxylation, which is necessary for de novo synthesis of amino acids, was decreased and affected the level of glutamine in hippocampal formation and those of glutamate, glutamine, GABA, and aspartate in the retrosplenial/cingulate cortex. Metabolic alterations were also detected in the entorhinal cortex. Overall, perturbations in energy- and neurotransmitter homeostasis, mitochondrial astrocytic and neuronal metabolism, and aspects of the glutamate-glutamine cycle were found in McGill-R-Thy1-APP rats.
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Axon-Sorting Multifunctional Nerve Guides: Accelerating Restoration of Nerve Function
2014-10-01
factor (singly & in selected combinations) in the organotypic model system for preferential sensory or motor axon extension. Use confocal microscopy to...track axon extension of labeled sensory or motor neurons from spinal cord slices (motor) or dorsal root ganglia ( DRG ) (sensory). 20 Thy1-YFP mice...RESEARCH ACCOMPLISHMENTS: • Established a system of color-coded mixed nerve tracking using GFP and RFP expressing motor and sensory neurons (Figure 1
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Ma, Ying; Shaik, Mohammed A; Kozberg, Mariel G; Kim, Sharon H; Portes, Jacob P; Timerman, Dmitriy; Hillman, Elizabeth M C
2016-12-27
Brain hemodynamics serve as a proxy for neural activity in a range of noninvasive neuroimaging techniques including functional magnetic resonance imaging (fMRI). In resting-state fMRI, hemodynamic fluctuations have been found to exhibit patterns of bilateral synchrony, with correlated regions inferred to have functional connectivity. However, the relationship between resting-state hemodynamics and underlying neural activity has not been well established, making the neural underpinnings of functional connectivity networks unclear. In this study, neural activity and hemodynamics were recorded simultaneously over the bilateral cortex of awake and anesthetized Thy1-GCaMP mice using wide-field optical mapping. Neural activity was visualized via selective expression of the calcium-sensitive fluorophore GCaMP in layer 2/3 and 5 excitatory neurons. Characteristic patterns of resting-state hemodynamics were accompanied by more rapidly changing bilateral patterns of resting-state neural activity. Spatiotemporal hemodynamics could be modeled by convolving this neural activity with hemodynamic response functions derived through both deconvolution and gamma-variate fitting. Simultaneous imaging and electrophysiology confirmed that Thy1-GCaMP signals are well-predicted by multiunit activity. Neurovascular coupling between resting-state neural activity and hemodynamics was robust and fast in awake animals, whereas coupling in urethane-anesthetized animals was slower, and in some cases included lower-frequency (<0.04 Hz) hemodynamic fluctuations that were not well-predicted by local Thy1-GCaMP recordings. These results support that resting-state hemodynamics in the awake and anesthetized brain are coupled to underlying patterns of excitatory neural activity. The patterns of bilaterally-symmetric spontaneous neural activity revealed by wide-field Thy1-GCaMP imaging may depict the neural foundation of functional connectivity networks detected in resting-state fMRI.
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Ma, Ying; Shaik, Mohammed A.; Kozberg, Mariel G.; Portes, Jacob P.; Timerman, Dmitriy
2016-01-01
Brain hemodynamics serve as a proxy for neural activity in a range of noninvasive neuroimaging techniques including functional magnetic resonance imaging (fMRI). In resting-state fMRI, hemodynamic fluctuations have been found to exhibit patterns of bilateral synchrony, with correlated regions inferred to have functional connectivity. However, the relationship between resting-state hemodynamics and underlying neural activity has not been well established, making the neural underpinnings of functional connectivity networks unclear. In this study, neural activity and hemodynamics were recorded simultaneously over the bilateral cortex of awake and anesthetized Thy1-GCaMP mice using wide-field optical mapping. Neural activity was visualized via selective expression of the calcium-sensitive fluorophore GCaMP in layer 2/3 and 5 excitatory neurons. Characteristic patterns of resting-state hemodynamics were accompanied by more rapidly changing bilateral patterns of resting-state neural activity. Spatiotemporal hemodynamics could be modeled by convolving this neural activity with hemodynamic response functions derived through both deconvolution and gamma-variate fitting. Simultaneous imaging and electrophysiology confirmed that Thy1-GCaMP signals are well-predicted by multiunit activity. Neurovascular coupling between resting-state neural activity and hemodynamics was robust and fast in awake animals, whereas coupling in urethane-anesthetized animals was slower, and in some cases included lower-frequency (<0.04 Hz) hemodynamic fluctuations that were not well-predicted by local Thy1-GCaMP recordings. These results support that resting-state hemodynamics in the awake and anesthetized brain are coupled to underlying patterns of excitatory neural activity. The patterns of bilaterally-symmetric spontaneous neural activity revealed by wide-field Thy1-GCaMP imaging may depict the neural foundation of functional connectivity networks detected in resting-state fMRI. PMID:27974609
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Davari, Maliheh; Soheili, Zahra-Soheila; Samiei, Shahram; Sharifi, Zohreh; Pirmardan, Ehsan Ranaei
2017-01-29
miR-183 cluster, composed of miR-183/-96/-182 genes, is highly expressed in the adult retina, particularly in photoreceptors. It involves in development, maturation and normal function of neuroretina. Ectopic overexpression of miR-183/-96/-182 genes was performed to assess reprogramming of hRPE cells. They were amplified from genomic DNA and cloned independently or in tandem configuration into pAAV.MCS vector. hRPE cells were then transfected with the recombinant constructs. Real-Time PCR was performed to measure the expression levels of miR-183/-96/-182 and that of several retina-specific neuronal genes such as OTX2, NRL, PDC and DCT. The transfected cells also were immunocytochemically examined for retina-specific neuronal markers, including Rhodopsin, red opsin, CRX, Thy1, CD73, recoverin and PKCα, to determine the cellular fate of the transfected hRPE cells. Data showed that upon miR-183/-96/-182 overexpression in hRPE cultures, the expression of neuronal genes including OTX2, NRL, PDC and DCT was also upregulated. Moreover, miR-183 cluster-treated hRPE cells were immunoreactive for neuronal markers such as Rhodopsin, red opsin, CRX and Thy1. Both transcriptional and translational upregulation of neuronal genes in miR-183 cluster-treated hRPE cells suggests that in vitro overexpression of miR-183 cluster could trigger reprogramming of hRPE cells to retinal neuron fate. Copyright © 2016 Elsevier Inc. All rights reserved.
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Hayworth, C R; Rojas, J C; Gonzalez-Lima, F
2008-01-01
This is the first study using a reporter transgenic model to investigate the effects of an environmental toxin on the retina. Rotenone is a widely used pesticide that inhibits mitochondrial complex I and produces neurotoxicity. Previous studies demonstrated the time course and dose response of rotenone toxicity on retinal ganglion cells (RGC). However, previous analyses of rotenone-induced retinotoxicity provided little detail of the optic nerve axons and cellular pathology. These limitations were successfully surmounted by using a transgenic mouse line shown to express cyan fluorescent protein (CFP) in neurons, including RGC, under regulatory elements of the human the thy1.1 promoter (thy-CFP). Data showed that CFP expression is limited to RGC and their processes in the retina of thy-CFP mice. Eyes exposed to the pesticide rotenone displayed marked alterations in RGC morphology, inner plexiform layer, optic disc, and optic nerves. After 24 h, the number of CFP-labeled RGC was reduced 50%. Correlated with a loss of RGC bodies was an approximate 50% reduction in CFP fluorescence intensity at the optic disc. The findings showed that rotenone-induced degeneration of RGC and their processes can be visualized with exquisite detail in thy-CFP mice, and that this approach may provide a novel and effective way to monitor the association between environmental toxins and neurodegeneration in living animals.
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Hu, Jinwei; Xu, Jane; Streelman, Matthew; Xu, Helen; Guthrie, O'neil
2015-01-01
Objective. The mechanisms of tinnitus are known to alter neuronal circuits in the brainstem and cortex, which are common to several comorbid conditions. This study examines the relationship between tinnitus and anxiety/depression. Subjects and Methods. Ninety-one male veterans with subjective tinnitus were enrolled in a Veterans Affairs Tinnitus Clinic. The Tinnitus Handicap Inventory (THI) was used to assess tinnitus severity. ICD-9 codes for anxiety/depression were used to determine their prevalence. Pure tone averages (PTA) were used to assess hearing status. Results. Descriptive analyses revealed that 79.1% of the 91 tinnitus sufferers had a diagnosis of anxiety, 59.3% had depression, and 58.2% suffered from both anxiety/depression. Patients with anxiety had elevated total THI scores as compared to patients without anxiety (p < 0.05). Patients with anxiety or depression had significantly increased Functional and Emotional THI scores, but not Catastrophic THI score. Significant positive correlations were illustrated between the degree of tinnitus and anxiety/depression (p < 0.05). There were no differences in PTA among groups. Conclusions. A majority of patients with tinnitus exhibited anxiety and depression. These patients suffered more severe tinnitus than did patients without anxiety and depression. The data support the need for multidisciplinary intervention of veterans with tinnitus. PMID:26697070
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Lindsey, James D; Duong-Polk, Karen X; Dai, Yi; Nguyen, Duy H; Leung, Christopher K; Weinreb, Robert N
2013-01-01
Thy-1 is a cell surface protein that is expressed during the differentiation of retinal ganglion cells (RGCs). Optic nerve injury induces progressive loss in the number of RGCs expressing Thy-1. The rate of this loss is fastest during the first week after optic nerve injury and slower in subsequent weeks. This study was undertaken to determine whether oral treatment with a water-soluble N-hydroxy-2,2,6,6-tetramethylpiperidine derivative (OT-440) protects against loss of Thy-1 promoter activation following optic nerve crush and whether this effect targets the earlier quick phase or the later slow phase. The retina of mice expressing cyan fluorescent protein under control of the Thy-1 promoter (Thy1-CFP mice) was imaged using a blue-light confocal scanning laser ophthalmoscope (bCSLO). These mice then received oral OT-440 prepared in cream cheese or dissolved in water, or plain vehicle, for two weeks and were imaged again prior to unilateral optic nerve crush. Treatments and weekly imaging continued for four more weeks. Fluorescent neurons were counted in the same defined retinal areas imaged at each time point in a masked fashion. When the counts at each time point were directly compared, the numbers of fluorescent cells at each time point were greater in the animals that received OT-440 in cream cheese by 8%, 27%, 52% and 60% than in corresponding control animals at 1, 2, 3 and 4 weeks after optic nerve crush. Similar results were obtained when the vehicle was water. Rate analysis indicated the protective effect of OT-440 was greatest during the first two weeks and was maintained in the second two weeks after crush for both the cream cheese vehicle study and water vehicle study. Because most of the fluorescent cells detected by bCSLO are RGCs, these findings suggest that oral OT-440 can either protect against or delay early degenerative responses occurring in RGCs following optic nerve injury.
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The Thiamine Biosynthesis Gene THI1 Promotes Nodule Growth and Seed Maturation1
Nagae, Miwa; Kawaguchi, Masayoshi; Takeda, Naoya
2016-01-01
Thiamine (vitamin B1) is essential for living organisms. Unlike animals, plants can synthesize thiamine. In Lotus japonicus, the expression of two thiamine biosynthesis genes, THI1 and THIC, was enhanced by inoculation with rhizobia but not by inoculation with arbuscular mycorrhizal fungi. THIC and THI2 (a THI1 paralog) were expressed in uninoculated leaves. THI2-knockdown plants and the transposon insertion mutant thiC had chlorotic leaves. This typical phenotype of thiamine deficiency was rescued by an exogenous supply of thiamine. In wild-type plants, THI1 was expressed mainly in roots and nodules, and the thi1 mutant had green leaves even in the absence of exogenous thiamine. THI1 was highly expressed in actively dividing cells of nodule primordia. The thi1 mutant had small nodules, and this phenotype was rescued by exogenous thiamine and by THI1 complementation. Exogenous thiamine increased nodule diameter, but the level of arbuscular mycorrhizal colonization was unaffected in the thi1 mutant or by exogenous thiamine. Expression of symbiotic marker genes was induced normally, implying that mainly nodule growth was delayed in the thi1 mutant. Furthermore, this mutant formed many immature seeds with reduced seed weight. These results indicate that thiamine biosynthesis mediated by THI1 enhances nodule enlargement and is required for seed development in L. japonicus. PMID:27702844
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Adrenoceptors in Brain: Cellular Gene Expression and Effects on Astrocytic Metabolism and [Ca2+]i
Hertz, Leif; Lovatt, Ditte; Goldman, Steven A.; Nedergaard, Maiken
2010-01-01
Recent in vivo studies have established astrocytes as a major target for locus coeruleus activation (Bekar et al., Cereb. Cortex 18, 2789–2795), renewing interest in cell culture studies on noradrenergic effects on astrocytes in primary cultures and calling for additional information about the expression of adrenoceptor subtypes on different types of brain cells. In the present communication, mRNA expression of α1-, α2- and β-adrenergic receptors and their subtypes was determined in freshly-isolated, cell marker-defined populations of astrocytes, NG2-positive cells, microglia, endothelial cells, and Thy1-positive neurons (mainly glutamatergic projection neurons) in murine cerebral cortex. Immediately after dissection of frontal, parietal and occipital cortex of 10–12-week-old transgenic mice, which combined each cell-type marker with a specific fluorescent signal, the tissue was digested, triturated and centrifuged, yielding a solution of dissociated cells of all types, which were separated by fluorescence-activated cell sorting (FACS). mRNA expression in each cell fraction was determined by microarray analysis. α1A-Receptors were unequivocally expressed in astrocytes and NG2-positive cells, but absent in other cell types, and α1B-receptors were not expressed in any cell population. Among α2-receptors only α2A-receptors were expressed, unequivocally in astrocytes and NG-positive cells, tentatively in microglia and questionably in Thy1-positive neurons and endothelial cells. β1-Receptors were unequivocally expressed in astrocytes, tentatively in microglia, and questionably in neurons and endothelial cells, whereas β2-adrenergic receptors showed tentative expression in neurons and astrocytes and unequivocal expression in other cell types. This distribution was supported by immunochemical data and its relevance established by previous studies in well-differentiated primary cultures of mouse astrocytes, showing that stimulation of α2-adrenoceptors increases glycogen formation and oxidative metabolism, the latter by a mechanism depending on intramitochondrial Ca2+, whereas α1-adrenoceptor stimulation enhances glutamate uptake, and β-adrenoceptor activation causes glycogenolysis and increased Na+,K+-ATPase activity. The Ca2+- and cAMP-mediated association between energy-consuming and energy-yielding processes is emphasized. PMID:20380860
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Effect of the GPI anchor of human Thy-1 on antibody recognition and function
Bradley, John E.; Chan, Joy M.; Hagood, James S.
2012-01-01
Thymocyte differentiation antigen 1 (Thy-1) is a glycosylphosphatidyl inositol (GPI)-linked cell surface glycoprotein expressed on numerous cell types, which regulates signals affecting cell adhesion, migration, differentiation, and survival. In addition, Thy-1 has been detected in serum, cerebral spinal fluid, wound fluid from venous ulcers, synovial fluid from joints in rheumatoid arthritis and more recently urine. We previously detected Thy-1 in the conditioned media of cytokine-stimulated lung fibroblasts, suggesting that Thy-1 shedding may be a response to cellular stress. Soluble and membrane bound forms of Thy-1 from in vivo sources have been shown to be identical in size when deglycosylated, suggesting that soluble Thy-1 is separated from the diacyl glycerol portion of its GPI-anchor by hydrolysis within the GPI moiety. For Thy-1 and other GPI-anchored proteins, delipidation induces a stable change in conformation that manifests itself in a change in antibody affinity for soluble forms. Using epitope tagged recombinant soluble Thy-1, we report that widely available monoclonal antibodies to human Thy-1 are unable to detect soluble Thy-1 by immunoblotting. We reevaluated the Thy-1 that we previously reported in the conditioned media of normal human lung fibroblasts and found it to be entirely insoluble. These findings suggest that most Thy-1 reported in body fluids retains its GPI anchor and may be associated with membrane fragments or vesicles. This phenomenon should be considered in the generation of antibodies and controls for Thy-1 bioassays. Furthermore, the changes in Thy-1 conformation with delipidation, beyond affecting antibody affinity, likely affect the ligand affinity and biological function of soluble vs. released membrane-associated forms. PMID:23358110
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The Effect of Substrate Topography on Direct Reprogramming of Fibroblasts to Induced Neurons
Kulangara, Karina; Adler, Andrew F.; Wang, Hong; Chellappan, Malathi; Hammett, Ellen; Yasuda, Ryohei; Leong, Kam W.
2014-01-01
Cellular reprogramming holds tremendous potential for cell therapy and regenerative medicine. Recently, fibroblasts have been directly converted into induced neurons (iNs) by overexpression of the neuronal transcription factors Ascl1, Brn2 and Myt1L. Hypothesizing that cell-topography interactions could influence the fibroblast-to-neuron reprogramming process, we investigated the effects of various topographies on iNs produced by direct reprogramming. Final iN purity and conversion efficiency were increased on micrograting substrates. Neurite branching was increased on microposts and decreased on microgratings, with a simplified dendritic arbor characterized by the reduction of MAP2+ neurites. Neurite outgrowth increased significantly on various topographies. DNA microarray analysis detected 20 differentially expressed genes in iNs reprogrammed on smooth versus microgratings, and quantitative PCR (qPCR) confirmed the upregulation of Vip and downregulation of Thy1 and Bmp5 on microgratings. Electrophysiology and calcium imaging verified the functionality of these iNs. This study demonstrates the potential of applying topographical cues to optimize cellular reprogramming. PMID:24709523
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A microprobe for parallel optical and electrical recordings from single neurons in vivo.
LeChasseur, Yoan; Dufour, Suzie; Lavertu, Guillaume; Bories, Cyril; Deschênes, Martin; Vallée, Réal; De Koninck, Yves
2011-04-01
Recording electrical activity from identified neurons in intact tissue is key to understanding their role in information processing. Recent fluorescence labeling techniques have opened new possibilities to combine electrophysiological recording with optical detection of individual neurons deep in brain tissue. For this purpose we developed dual-core fiberoptics-based microprobes, with an optical core to locally excite and collect fluorescence, and an electrolyte-filled hollow core for extracellular single unit electrophysiology. This design provides microprobes with tips < 10 μm, enabling analyses with single-cell optical resolution. We demonstrate combined electrical and optical detection of single fluorescent neurons in rats and mice. We combined electrical recordings and optical Ca²(+) measurements from single thalamic relay neurons in rats, and achieved detection and activation of single channelrhodopsin-expressing neurons in Thy1::ChR2-YFP transgenic mice. The microprobe expands possibilities for in vivo electrophysiological recording, providing parallel access to single-cell optical monitoring and control.
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Zhou, Yong; Hagood, James S.; Murphy-Ullrich, Joanne E.
2004-01-01
Distinct subpopulations of fibroblasts contribute to lung fibrosis, although the mechanisms underlying fibrogenesis in these subpopulations are not clear. Differential expression of the glycophosphatidylinositol-linked protein Thy-1 affects proliferation and myofibroblast differentiation. Lung fibroblast populations selected on the basis of Thy-1 expression by cell sorting were examined for responses to fibrogenic stimuli. Thy-1 (−) and Thy-1 (+) fibroblast populations were treated with platelet-derived growth factor-BB, interleukin-1β, interleukin-4, or bleomycin and assessed for activation of transforming growth factor (TGF)-β, Smad3 phosphorylation, and α-smooth muscle actin and fibronectin expression. Thy-1 (−) fibroblasts responded to these stimuli with increased TGF-β activity, Smad3 phosphorylation, and expression of α-smooth muscle actin and fibronectin, whereas Thy-1 (+) fibroblasts resisted stimulation. The unresponsiveness of Thy-1 (+) cells is not because of defective TGF-β signaling because both subsets respond to exogenous active TGF-β. Rather, Thy-1 (−) fibroblasts activate latent TGF-β in response to fibrogenic stimuli, whereas Thy-1 (+) cells fail to do so. Defective activation is common to multiple mechanisms of TGF-β activation, including thrombospondin 1, matrix metalloproteinase, or plasmin. Thy-1 (−) lung fibroblasts transfected with Thy-1 also become resistant to fibrogenic stimulation, indicating that Thy-1 is a critical biological response modifier that protects against fibrotic progression by controlling TGF-β activation. These studies provide a molecular basis for understanding the differential roles of fibroblast subpopulations in fibrotic lung disease through control of latent TGF-β activation. PMID:15277239
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Lack of Thy1 (CD90) expression in neuroblastomas is correlated with impaired survival.
Fiegel, Henning C; Kaifi, Jussuf T; Quaas, Alexander; Varol, Emine; Krickhahn, Annika; Metzger, Roman; Sauter, Guido; Till, Holger; Izbicki, Jakob R; Erttmann, Rudolf; Kluth, Dietrich
2008-01-01
Neuroblastoma (NBL) is the most common solid tumor in children. Tumors in advanced stage or with positive risk factors still have a poor prognosis. Thy1 (CD90) is a membrane glycoprotein expressed in thymus, retinal ganglionic cells, and several types of stem cells. The aim of this study was to assess Thy1 expression in NBL and analyze the correlation with clinical outcome. Sixty-three specimens of NBL were stained for Thy1 on a tissue microarray by immunohistochemistry. Fresh frozen tumor tissues were used for RNA isolation, and RT-PCR analysis for Thy1-mRNA expression was performed. Patients' survival data were correlated with Thy1 status using a log rank test and a Cox regression multivariate analysis. Thy1 was expressed on 51 (81%) of the tumors. Kaplan-Meier survival analysis showed a significantly impaired survival in patients with NBL missing Thy1 (P < 0.005 by log-rank test). A multivariate Cox regression showed an independent prognostic value of Thy1 status for overall survival (P < 0.05). In addition, the frequency of events and deaths was significantly higher in the group of patients with Thy1 negative tumors, as assessed by ANOVA analysis (P < 0.05 by F-test). The data showed that Thy1-negative NBL patients have a significantly impaired overall survival compared with Thy1-positive NBL patients. Thus, Thy1 seemed to be a marker with a specific prognostic value in NBL patients. Future studies are aiming at the biological role of this marker in the tumor cell differentiation.
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Horiguchi, Kotaro; Nakakura, Takashi; Yoshida, Saishu; Tsukada, Takehiro; Kanno, Naoko; Hasegawa, Rumi; Takigami, Shu; Ohsako, Shunji; Kato, Takako; Kato, Yukio
2016-11-11
Contact-dependent (juxtacrine) signaling is important for local cell-to-cell interaction and has received attention in recent years regarding its role in pituitary function, differentiation, and development. This study investigated one of the juxtacrine-related molecules, thymocyte differentiation antigen 1 (THY1), in the anterior lobe of the rat pituitary gland. Western blot analysis revealed expression of the THY1 protein in the adult rat anterior lobe. We also found that the THY1 ligand, integrin-β2 (ITGB2), is also expressed in the pituitary gland. In situ hybridization and immunohistochemical analyses showed that both THY1 mRNA and protein were present in almost, if not all, thyroid-stimulating hormone (TSH)-immunopositive cells (thyrotropes) and that ITGB2 was co-expressed in these cells. As THY1 appeared to represent a novel marker for thyrotropes, we then attempted to isolate these cells from various anterior lobe cells by the use of a THY1 antibody and a pluriBead-cascade cell isolation system. This technology allowed the isolation of thyrotropes with 83% purity at about 17-fold enrichment. Furthermore, the isolated THY1-immunopositive cells had higher Tsh mRNA levels compared with THY1-immunonegative cells and released TSH in response to thyrotropin-releasing hormone. These findings indicated that THY1 represents a potent thyrotrope marker and that the thyrotrope isolation method using the THY1 antibody may serve as a powerful tool to analyze their function including juxtacrine regulation through THY1/ITGB2 interaction. Copyright © 2016 Elsevier Inc. All rights reserved.
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Eleftheriadou, I; Trabalza, A; Ellison, SM; Gharun, K; Mazarakis, ND
2014-01-01
To understand how receptors are involved in neuronal trafficking and to be able to utilize them for specific targeting via the peripheral route would be of great benefit. Here, we describe the generation of novel lentiviral vectors with tropism to motor neurons that were made by coexpressing onto the lentiviral surface a fusogenic glycoprotein (mutated sindbis G) and an antibody against a cell-surface receptor (Thy1.1, p75NTR, or coxsackievirus and adenovirus receptor) on the presynaptic terminal of the neuromuscular junction. These vectors exhibit binding specificity and efficient transduction of receptor positive cell lines and primary motor neurons in vitro. Targeting of each of these receptors conferred to these vectors the capability of being transported retrogradely from the axonal tip, leading to transduction of motor neurons in vitro in compartmented microfluidic cultures. In vivo delivery of coxsackievirus and adenovirus receptor-targeted vectors in leg muscles of mice resulted in predicted patterns of motor neuron labeling in lumbar spinal cord. This opens up the clinical potential of these vectors for minimally invasive administration of central nervous system-targeted therapeutics in motor neuron diseases. PMID:24670531
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Giusti, Massimo; Massa, Barbara; Balestra, Margherita; Calamaro, Paola; Gay, Stefano; Schiaffino, Simone; Turtulici, Giovanni; Zupo, Simonetta; Monti, Eleonora; Ansaldo, Gianluca
2017-01-01
The cytology of 130 indeterminate nodules (Thy 3) was retrospectively reviewed according to the British Thyroid Association 2014 classification. Nodules were divided into Thy 3a (atypical features) and Thy 3f (follicular lesion) categories. Histology was available as a reference for 97 nodules. Pre-surgical evaluations comprised biochemical tests, color-Doppler ultrasonography (US), semi-quantitative elastography-US (USE), contrast-enhanced US (CEUS), and mutation analysis from cytological slides. Thyroid malignancy was the final diagnosis for 19% of surgically-treated nodules. No statistically significant difference in the risk of malignancy was found between Thy 3a (26%) and Thy 3f (14%) nodules. Histology of the Thy 3a and Thy 3f nodules showed a higher incidence of Hurtle cell adenomas in Thy 3f (29%) than in Thy 3a (3%) nodules (P=0.01). The only pre-surgical difference concerned the BRAF V600E mutation, which was positive in some Thy 3a but not in any Thy 3f nodules (P=0.04). Receiver-operating characteristic (ROC) analysis was used to obtain cut-off values from US (score), USE (ELX 2/1 strain index), and CEUS (time-to-peak index and peak index) data. The cut-off values were similar for Thy 3a and Thy 3f nodules. Data showed that malignancy can be suspected if the US score is >2, ELX 1/2 strain index >1, time-to-peak index >1, and peak index <1. In a sub-group of 24 revised nodules (12 Thy 3a and 12 Thy 3f) with histology as a reference, the diagnostic power of cumulative pre-surgical analysis by means of US, USE, and CEUS showed high positive and negative predictive values (83% and 100%, respectively) for the presence of malignancy in Thy 3a and Thy 3f nodules. In conclusion, in our series of revised Thy 3 nodules, malignancy was low and displayed no significant differences between Thy 3a and Thy 3f categories. The use of cut-offs based on histology as a reference could reduce surgery. Our data support the conviction that, in mutation-negative Thy 3a and Thy 3f nodules, observation should be the first choice when not all instrumental results are suspect. PMID:28681580
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Viereckel, Thomas; Dumas, Sylvie; Smith-Anttila, Casey J. A.; Vlcek, Bianca; Bimpisidis, Zisis; Lagerström, Malin C.; Konradsson-Geuken, Åsa; Wallén-Mackenzie, Åsa
2016-01-01
The ventral tegmental area (VTA) and substantia nigra pars compacta (SNc) of the midbrain are associated with Parkinson’s disease (PD), schizophrenia, mood disorders and addiction. Based on the recently unraveled heterogeneity within the VTA and SNc, where glutamate, GABA and co-releasing neurons have been found to co-exist with the classical dopamine neurons, there is a compelling need for identification of gene expression patterns that represent this heterogeneity and that are of value for development of human therapies. Here, several unique gene expression patterns were identified in the mouse midbrain of which NeuroD6 and Grp were expressed within different dopaminergic subpopulations of the VTA, and TrpV1 within a small heterogeneous population. Optogenetics-coupled in vivo amperometry revealed a previously unknown glutamatergic mesoaccumbal pathway characterized by TrpV1-Cre-expression. Human GRP was strongly detected in non-melanized dopaminergic neurons within the SNc of both control and PD brains, suggesting GRP as a marker for neuroprotected neurons in PD. This study thus unravels markers for distinct subpopulations of neurons within the mouse and human midbrain, defines unique anatomical subregions within the VTA and exposes an entirely new glutamatergic pathway. Finally, both TRPV1 and GRP are implied in midbrain physiology of importance to neurological and neuropsychiatric disorders. PMID:27762319
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Cajal's second great battle for the neuron doctrine: the nature and function of neurofibrils.
Frixione, Eugenio
2009-03-01
One hundred years ago, a novel kind of reticularism threatened to displace the neuron doctrine as the established model of functional organization of the nervous system. The challenging paradigm, championed by Stephan von Apáthy and Albrecht Bethe, held that nerve impulses propagate along neurofibrils connected in a continuous network throughout all nerve cells. Santiago Ramón y Cajal, a leading figure in the conception of the neuron doctrine, headed again the battle against this return of reticularism. Dissatisfied with the available staining techniques, he devised the "reduced silver nitrate method" that even Camillo Golgi recognized as the best at the time for revealing the neurofibrils. In 1904 Cajal already published over a dozen papers in three languages describing neurofibril distributions in the nervous systems of diverse vertebrates and invertebrates, under both normal and experimental conditions. Next he investigated the involvement of neurofibrils in the process of nerve regeneration. This unprecedented survey led him to the conclusion that the neurofibrils are linear "colonies" of particles constituting a semi-solid, dynamic internal skeleton of the nerve cell. Apáthy reacted with a long invective paper that Cajal had no choice but acknowledging. His comprehensive reply, published in 1908, meant the effective end of the renewed reticularist campaign against the neuron doctrine. Along the way, a visionary and today almost forgotten chapter in the history of the cytoskeleton had also been written.
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Thy-1+ dendritic epidermal cells express T3 antigen and the T-cell receptor gamma chain.
Stingl, G; Koning, F; Yamada, H; Yokoyama, W M; Tschachler, E; Bluestone, J A; Steiner, G; Samelson, L E; Lew, A M; Coligan, J E
1987-01-01
The murine epidermis is a heterogeneous epithelium composed of keratinocytes, melanocytes, Langerhans cells, and a recently described subpopulation (2-3%) of bone-marrow-derived leukocytes with a dendritic morphology and the cell surface phenotype Thy-1+, L3T4-, Lyt-2-. Previous studies have demonstrated that cell lines derived from freshly explanted Thy-1+ dendritic epidermal cells (DEC) have abundant mRNA for rearranged T-cell receptor (TCR) gamma-chain genes. Analysis of Thy-1+ DEC in situ, freshly isolated cell suspensions of Thy-1+ DEC, and long-term Thy-1+ DEC lines demonstrated that 100% of the Thy-1+ DEC reacted with a monoclonal antibody to the epsilon chain of the murine T3 complex and that 40-60% of resident Thy-1+ DEC were also reactive with an antiserum to the TCR gamma chain. Two Thy-1+ DEC lines expressed a disulfide-linked 70-kDa molecule that could be precipitated with an anti-gamma-chain antiserum and could be coprecipitated with an antiserum to the T3 delta chain; the molecule appeared as a single 34-kDa band under reducing conditions. The phenotype of Thy-1+ DEC (T3+, L3T4-, Lyt-2-, TCR gamma chain+) thus resembles that of the recently described subpopulation of murine and human lymphocytes that have been identified in the thymus, peripheral blood, and fetal blood. Images PMID:2885839
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Yuan, Yin; Xu, Xiu-yue; Lao, Jie; Zhao, Xin
2018-01-01
Nerve transfer is the most common treatment for total brachial plexus avulsion injury. After nerve transfer, the movement of the injured limb may be activated by certain movements of the healthy limb at the early stage of recovery, i.e., trans-hemispheric reorganization. Previous studies have focused on functional magnetic resonance imaging and changes in brain-derived neurotrophic factor and growth associated protein 43, but there have been no proteomics studies. In this study, we designed a rat model of total brachial plexus avulsion injury involving contralateral C7 nerve transfer. Isobaric tags for relative and absolute quantitation and western blot assay were then used to screen differentially expressed proteins in bilateral motor cortices. We found that most differentially expressed proteins in both cortices of upper limb were associated with nervous system development and function (including neuron differentiation and development, axonogenesis, and guidance), microtubule and cytoskeleton organization, synapse plasticity, and transmission of nerve impulses. Two key differentially expressed proteins, neurofilament light (NFL) and Thy-1, were identified. In contralateral cortex, the NFL level was upregulated 2 weeks after transfer and downregulated at 1 and 5 months. The Thy-1 level was upregulated from 1 to 5 months. In the affected cortex, the NFL level increased gradually from 1 to 5 months. Western blot results of key differentially expressed proteins were consistent with the proteomic findings. These results indicate that NFL and Thy-1 play an important role in trans-hemispheric organization following total brachial plexus root avulsion and contralateral C7 nerve transfer. PMID:29557385
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Ohtoyo, Mamoru; Machinaga, Nobuo; Inoue, Ryotaku; Hagihara, Katsunobu; Yuita, Hiroshi; Tamura, Masakazu; Hashimoto, Ryuji; Chiba, Jun; Muro, Fumihito; Watanabe, Jun; Kobayashi, Yoshimasa; Abe, Koji; Kita, Yasuo; Nagasaki, Miyuki; Shimozato, Takaichi
2016-05-19
Caramel color is widely used in the food industry, and its many variations are generally considered to be safe. It has been known for a long time that THI (2-acetyl-4-(tetrahydroxybutyl)imidazole), a component of caramel color III, causes lymphopenia in animals through sphingosine 1-phosphate (S1P) lyase (S1PL) inhibition. However, this mechanism of action has not been fully validated because THI does not inhibit S1PL in vitro. To reconcile this situation, we examined molecular details of THI mechanism of action using "smaller" THI derivatives. We identified a bioactive derivative, A6770, which has the same lymphopenic effect as THI via S1PL inhibition. In the case of A6770 we observe this effect both in vitro and in vivo, and demonstrate that A6770 is phosphorylated and inhibits S1PL in the same way as 4-deoxypyridoxine. In addition, A6770 was detected in rat plasma following oral administration of THI, suggesting that A6770 is a key metabolic intermediate of THI. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Park, Eun Jung; Jang, Hwa Jin; Tsoyi, Konstantin; Kim, Young Min; Park, Sang Won; Kim, Hye Jung; Lee, Jae Heun; Chang, Ki Churl
2013-01-01
The nuclear DNA binding protein high mobility group box 1 (HMGB1) has recently been suggested to act as a late mediator of septic shock. The effect of ((S)-6,7-dihydroxy-1-(4-hydroxynaphthylmethyl)-1,2,3,4-tetrahydroisoquinoline alkaloid, also known as THI-56, in an experimental model of sepsis was investigated. THI-56 exhibited potent anti-inflammatory properties in response to LPS in RAW 264.7 cells. In particular, THI-56 significantly inhibited the expression of inducible nitric oxide synthase (iNOS) and the release of HMGB1 in activated macrophages. THI-56 activated NE-F2-regulated factor 2 (Nrf-2)/heme oxygenase 1 (HO-1). The specific knockdown of the HO-1 gene by HO-1 siRNA significantly reversed the inhibitory effects of THI-56 on iNOS expression and HMGB1 release in LPS-stimulated macrophages. Importantly, THI-56 administration protected animals from death induced by either a lethal dose of LPS or cecal ligation and puncture (CLP). Furthermore, the ALT, AST, BUN, creatinine, and HMGB1 levels in the blood were significantly increased in CLP-induced septic mice, and the administration of THI-56 reduced these levels in a concentration-dependent and zinc protoporphyrin IX (ZnPPIX)-sensitive manner. In addition, the administration of THI-56 significantly ameliorated not only lung damage but also macrophage infiltration in the livers of CLP-induced septic mice, and these effects were also abrogated in the presence of ZnPPIX. Thus, we conclude that THI-56 significantly attenuates the proinflammatory response induced by LPS and reduces organ damage in a CLP-induced sepsis model through the upregulation of Nrf-2/HO-1.
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Cell Surface THY-1 Contributes to Human Cytomegalovirus Entry via a Macropinocytosis-Like Process
Li, Qingxue; Fischer, Elizabeth
2016-01-01
ABSTRACT Previously we showed that THY-1 has a critical role in the initial stage of infection of certain cell types with human cytomegalovirus (HCMV) and that THY-1 is important for HCMV-mediated activation of phosphatidylinositol 3-kinase (PI3K)/Akt during virus entry. THY-1 is known to interact with integrins and is a major cargo protein of clathrin-independent endocytic vesicles. Since macropinocytosis involves integrin signaling, is PI3K/Akt dependent, and is a clathrin-independent endocytic process, we determined whether THY-1 has a role in HCMV entry by macropinocytosis. Using electron microscopy in two cell lines that support HCMV infection in a THY-1-dependent manner, we found that HCMV enters these cells by a macropinocytosis-like process. THY-1 associated with HCMV virions on the cell surface and colocalized with virus inside macropinosomes. 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) and soluble THY-1 blocked HCMV infection in the cell lines by ≥80% and 60%, respectively. HCMV entry into the cells triggered increased influx of extracellular fluid, a marker of macropinocytosis, and this increased fluid uptake was inhibited by EIPA and by soluble THY-1. Blocking actin depolymerization, Na+/H+ exchange, PI3K, and Pak1 kinase, which are critical for macropinocytosis, impaired HCMV infection. Neither internalized HCMV virions nor THY-1 in virus-infected cells colocalized with transferrin as determined by confocal microscopy, indicating that clathrin-mediated endocytosis was not involved in THY-1-associated virus entry. These results suggest that HCMV has adapted to utilize THY-1, a cargo protein of clathrin-independent endocytotic vesicles, to facilitate efficient entry into certain cell types by a macropinocytosis-like process. IMPORTANCE Human cytomegalovirus (HCMV) infects over half of the population and is the most common infectious cause of birth defects. The virus is the most important infection occurring in transplant recipients. The mechanism of how HCMV enters cells is controversial. In this study, we show that THY-1, a cell surface protein that is critical for the early stage of entry of HCMV into certain cell types, contributes to virus entry by macropinocytosis. Our findings suggest that HCMV has adapted to utilize THY-1 to facilitate entry of HCMV into macropinosomes in certain cell types. Further knowledge about the mechanism of HCMV entry into cells may facilitate the development of novel inhibitors of virus infection. PMID:27558416
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Improved expression of halorhodopsin for light-induced silencing of neuronal activity
Zhao, Shengli; Cunha, Catarina; Zhang, Feng; Liu, Qun; Gloss, Bernd; Deisseroth, Karl; Augustine, George J.; Feng, Guoping
2011-01-01
The ability to control and manipulate neuronal activity within an intact mammalian brain is of key importance for mapping functional connectivity and for dissecting the neural circuitry underlying behaviors. We have previously generated transgenic mice that express channelrhodopsin-2 for light-induced activation of neurons and mapping of neural circuits. Here we describe transgenic mice that express halorhodopsin (NpHR), a light-driven chloride pump, that can be used to silence neuronal activity via light. Using the Thy-1 promoter to target NpHR expression to neurons, we found that neurons in these mice expressed high levels of NpHR-YFP and that illumination of cortical pyramidal neurons expressing NpHR-YFP led to rapid, reversible photoinhibition of action potential firing in these cells. However, NpHR-YFP expression led to the formation of numerous intracellular blebs, which may disrupt neuronal function. Labeling of various subcellular markers indicated that the blebs arise from retention of NpHR-YFP in the endoplasmic reticulum. By improving the signal peptide sequence and adding an ER export signal to NpHR-YFP, we eliminated the formation of blebs and dramatically increased the membrane expression of NpHR-YFP. Thus, the improved version of NpHR should serve as an excellent tool for neuronal silencing in vitro and in vivo. PMID:18931914
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Assmann, Karel J M; van Son, Jacco P H F; Dïjkman, Henry B P M; Mentzel, Stef; Wetzels, Jack F M
2002-07-01
Podocytes play an important role in the development of proteinuria and focal glomerulosclerosis. Previously we have demonstrated that a combination of two monoclonal antibodies (mAb) against aminopeptidase A (APA), an enzyme present on podocytes, induces a massive acute albuminuria in mice. The present study examined the relationship between the acute antibody-induced albuminuria and the development of focal glomerulosclerosis in the Thy-1.1 transgenic mouse. This mouse expresses a hybrid human-mouse Thy-1.1 antigen on the podocytes, and slowly but spontaneously develops albuminuria and focal glomerulosclerosis. Five-week-old non-albuminuric Thy-1.1 transgenic and non-transgenic control mice were injected with anti-APA and anti-Thy-1.1 mAb or saline. Albuminuria was measured at days 1, 7, 14 and 21. At day 21 kidneys were processed for light microscopy, immunofluorescence, and electron microscopy. Injection of anti-APA and anti-Thy1.1 mAb in Thy-1.1 transgenic mice induced an albuminuria at day 1 that persisted at day 21. The acute albuminuria after injection of anti-APA mAb was more prominent but transient in non-transgenic mice. In non-trangenic mice no albuminuria could be induced with anti-Thy 1.1 mAb. Light microscopy revealed normal glomeruli at day 1 in all transgenic mice, however, at day 21 advanced glomerulosclerotic lesions were seen in mice injected with either anti-APA mAb (37+/-19% of glomeruli affected) or anti-Thy-1.1 mAb (71+/-5%). Non-transgenic mice did not reveal sclerotic lesions at any time investigated. In the transgenic mice the percentage of focal glomerulosclerosis at day 21 did not correlate with albuminuria at day 21. However, we found a highly significant correlation between percentage of focal glomerulosclerosis and the time-averaged albuminuria over the three-week study period (P < 0.001). Injection of a combination of anti-APA or anti-Thy-1.1 mAb into one mo old, non-albuminuric Thy-1.1 transgenic mice induces an acute albuminuria at day 1 that is accompanied by an accelerated focal glomerulosclerosis at day 21. We suggest that the Thy-1.1 transgenic mouse is an excellent model to study specifically the relation between podocytic injury, albuminuria and the development of focal glomerulosclerosis.
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Visualizing estrogen receptor-a-expressing neurons using a new ERa-ZsGreen reporter mouse line
USDA-ARS?s Scientific Manuscript database
A variety of biological functions of estrogens, including regulation of energy metabolism, are mediated by neurons expressingestrogen receptor-a (ERa) in the brain. However, complex intracellular processes in these ERa-expressing neurons are difficult to unravel, due to the lack of strategy to visua...
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Structural genes for thiamine biosynthetic enzymes (thiCEFGH) in Escherichia coli K-12.
Vander Horn, P B; Backstrom, A D; Stewart, V; Begley, T P
1993-01-01
Escherichia coli K-12 synthesizes thiamine pyrophosphate (vitamin B1) de novo. Two precursors [4-methyl-5-(beta-hydroxyethyl)thiazole monophosphate and 4-amino-5-hydroxymethyl-2-methylpyrimidine pyrophosphate] are coupled to form thiamine monophosphate, which is then phosphorylated to make thiamine pyrophosphate. Previous studies have identified two classes of thi mutations, clustered at 90 min on the genetic map, which result in requirements for the thiazole or the hydroxymethylpryimidine. We report here our initial molecular genetic analysis of the thi cluster. We cloned the thi cluster genes and examined their organization, structure, and function by a combination of phenotypic testing, complementation analysis, polypeptide expression, and DNA sequencing. We found five tightly linked genes, designated thiCEFGH. The thiC gene product is required for the synthesis of the hydroxymethylpyrimidine. The thiE, thiF, thiG, and thiH gene products are required for synthesis of the thiazole. These mutants did not respond to 1-deoxy-D-threo-2-pentulose, indicating that they are blocked in the conversion of this precursor compound to the thiazole itself. Images PMID:8432721
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Preliminary characterization of Thy-1.1 and Ag-B antigens from rat tissues solubilized in detergents
Letarte-Muirhead, Michelle; Acton, Ronald T.; Williams, Alan F.
1974-01-01
1. A radioactive binding assay for Thy-1.1 alloantigen which functions in the presence of detergents was established by using glutaraldehyde-fixed thymocytes as target cells. Thy-1.1 activity in detergent extracts was then assayed by measuring inhibition of the binding assay. 2. Solubilization of Thy-1.1 from whole thymocytes, and their membranes by a large number of non-ionic detergents and deoxycholate was studied. In the same extracts Ag-B(4) histocompatibility antigenic activities were measured. With the exception of Nonidet P-40, the detergents did not affect the antigenicity of Thy-1.1, but only Lubrol-PX and deoxycholate gave effective solubilization as measured by activity remaining in the supernatant after centrifugation at 200000g for 40min. With Ag-B(4) antigen, Triton X-100, Triton X-67 and Nonidet P-40 gave effective solubilization as well as Lubrol-PX and deoxycholate. Solubilization of Thy-1.1 activity from leukaemia cells and a brain homogenate was also studied, but none of the non-ionic detergents gave satisfactory results with these tissues. 3. Extracts from thymocyte membranes were further examined by gel filtration and sucrose gradient centrifugation. The Thy-1.1 activity behaved as a single component in deoxycholate with a density similar to that of a globular protein, but in Lubrol-PX the antigen was contained in a low-density complex. In Lubrol-PX extracts Ag-B(4) was also found in aggregates not observed in deoxycholate. 4. The s20,w values for Thy-1.1 and Ag-B(4) antigens in deoxycholate were 2.4 and 4.4, and v̄ values were 0.70 and 0.75 respectively. The Stokes radius observed for Thy-1.1 was 3.1nm and for Ag-B(4) 5.3nm. By using these values the molecular weights for the antigen–detergent complexes were calculated to be 28000 for Thy-1.1 and 100000 for Ag-B(4). PMID:4219284
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Richter, Franziska; Gao, Fuying; Medvedeva, Vera; Lee, Patrick; Bove, Nicholas; Fleming, Sheila M.; Michaud, Magali; Lemesre, Vincent; Patassini, Stefano; De La Rosa, Krystal; Mulligan, Caitlin K.; Sioshansi, Pedrom; Zhu, Chunni; Coppola, Giovanni; Bordet, Thierry; Pruss, Rebecca; Chesselet, Marie-Françoise
2014-01-01
Cholesterol-oximes TRO19622 and TRO40303 target outer mitochondrial membrane proteins and have beneficial effects in preclinical models of neurodegenerative diseases leading to their advancement to clinical trials. Dopaminergic neurons degenerate in Parkinson’s disease (PD) and are prone to oxidative stress and mitochondrial dysfunction. In order to provide insights into the neuroprotective potential of TRO19622 and TRO40303 for dopaminergic neurons in vivo, we assessed their effects on gene expression in laser captured nigrostriatal dopaminergic neurons of wildtype mice and of mice that over-express alpha-synuclein, a protein involved in both familial and sporadic forms of PD (Thy1-aSyn mice). Young mice were fed the drugs in food pellets or a control diet from 1 to 4 months of age, approximately 10 months before the appearance of striatal dopamine loss in this model. Unbiased weighted gene co-expression network analysis (WGCNA) of transcriptional changes revealed effects of cholesterol oximes on transcripts related to mitochondria, cytoprotection and anti-oxidant response in wild-type and transgenic mice, including increased transcription of stress defense (e.g. Prdx1, Prdx2, Glrx2, Hspa9, Pink1, Drp1, Trak1) and dopamine-related (Th, Ddc, Gch1, Dat, Vmat2, Drd2, Chnr6a) genes. Even at this young age transgenic mice showed alterations in transcripts implicated in mitochondrial function and oxidative stress (e.g. Bcl-2, Bax, Casp3, Nos2), and both drugs normalized about 20% of these alterations. Young Thy1-aSyn mice exhibit motor deficits that differ from parkinsonism and are established before the onset of treatment; these deficits were not improved by cholesterol oximes. However, high doses of TRO40303 improved olfaction and produced the same effects as dopamine agonists on a challenging beam test, specifically an increase in footslips, an observation congruent with its effects on transcripts involved in dopamine synthesis. High doses of TRO19622 increased alpha-synuclein aggregates in the substantia nigra; this effect, not seen with TRO40303 was inconsistent and may represent a protective mechanism as in other neurodegenerative diseases. Overall, the results suggest that cholesterol oximes, while not improving early effects of alpha-synuclein overexpression on motor behavior or pathology, may ameliorate the function and resilience of dopaminergic neurons in vivo and support further studies of neuroprotection in models with dopaminergic cell loss. PMID:24844147
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Hepatic oval cells express the hematopoietic stem cell marker Thy-1 in the rat.
Petersen, B E; Goff, J P; Greenberger, J S; Michalopoulos, G K
1998-02-01
Hepatic oval cells (HOC) are a small subpopulation of cells found in the liver when hepatocyte proliferation is inhibited and followed by some type of hepatic injury. HOC can be induced to proliferate using a 2-acetylaminofluorene (2-AAF)/hepatic injury (i.e., CCl4, partial hepatectomy [PHx]) protocol. These cells are believed to be bipotential, i.e., able to differentiate into hepatocytes or bile ductular cells. In the past, isolation of highly enriched populations of these cells has been difficult. Thy-1 is a cell surface marker used in conjunction with CD34 and lineage-specific markers to identify hematopoietic stem cells. Thy-1 antigen is not normally expressed in adult liver, but is expressed in fetal liver, presumably on the hematopoietic cells. We report herein that HOC express high levels of Thy-1. Immunohistochemistry revealed that the cells expressing Thy-1 were indeed oval cells, because they also expressed alpha-fetoprotein (AFP), gamma-glutamyl transpeptidase (GGT), cytokeratin 19 (CK-19), OC.2, and OV-6, all known markers for oval cell identification. In addition, the Thy-1+ cells were negative for desmin, a marker specific for Ito cells. Using Thy-1 antibody as a new marker for the identification of oval cells, a highly enriched population was obtained. Using flow cytometric methods, we isolated a 95% to 97% pure Thy-1+ oval cell population. Our results indicate that cell sorting using Thy-1 could be an attractive tool for future studies, which would facilitate both in vivo and in vitro studies of HOC.
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Single-Cell Genomics Unravels Brain Cell-Type Complexity.
Guillaumet-Adkins, Amy; Heyn, Holger
2017-01-01
The brain is the most complex tissue in terms of cell types that it comprises, to the extent that it is still poorly understood. Single cell genome and transcriptome profiling allow to disentangle the neuronal heterogeneity, enabling the categorization of individual neurons into groups with similar molecular signatures. Herein, we unravel the current state of knowledge in single cell neurogenomics. We describe the molecular understanding of the cellular architecture of the mammalian nervous system in health and in disease; from the discovery of unrecognized cell types to the validation of known ones, applying these state-of-the-art technologies.
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Nabenishi, Hisashi; Ohta, Hiroshi; Nishimoto, Toshihumi; Morita, Tetsuo; Ashizawa, Koji; Tsuzuki, Yasuhiro
2011-09-01
In the present study, we investigated the relationship between the temperature-humidity index (THI) and the conception rate of lactating dairy cows in southwestern Japan, one of the hottest areas of the country. We also investigated the relationship between measurement of the vaginal temperature of lactating dairy cows as their core body temperature at one-hour intervals for 25 consecutive days in hot (August-September, n=6) and cool (January-February, n=5) periods and their THI. Furthermore, we discussed the above relationship using these vaginal temperatures, the conception rates and the THI. As a result, when the conception rates from day 2 to 0 before AI were classified into day 2, 1 and 0 groups by the six maximum THI values in each group (mTHI; <61, 61-65, 66-70, 71-75, 76-80, >80), only the conception rate for the mTHI over 80 at 1 day before AI group was significantly lower (P<0.05) than the other groups. The conception rate for days 15 to 17, but not days 19 to 22 and 30 to 35, after AI in the cows that experienced average mTHI over 80 (amTHI>80) was significantly lower (P<0.05) than that of the cows that did not experience amTHI>80. There was a significant positive correlation (P<0.01) between the mTHI and the mean daily vaginal temperature, but not during the cool period. When the mTHI reached 69, the vaginal temperature started to increase. As for the relationship between the conception rates and vaginal temperatures for all mTHI classes, in the mTHI>80 at 1 day before AI group, the vaginal temperature increased by 0.6 C from 38.7 C, resulting in a reduction of 11.6% in the conception rate from 40.5%. In conclusion, these results suggest that one of the causes of the fall in conception rate of lactating dairy cows during the summer season in southwestern Japan may be an increase in their core body temperature with a higher mTHI than the critical mTHI of 69 at 1 day before AI.
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Histone Deacetylase 3 Is Necessary for Proper Brain Development*
Norwood, Jordan; Franklin, Jade M.; Sharma, Dharmendra; D'Mello, Santosh R.
2014-01-01
The functional role of histone deacetylase 3 (HDAC3) in the developing brain has yet to be elucidated. We show that mice lacking HDAC3 in neurons and glia of the central nervous system, Nes-Cre/HDAC3 conditional KO mice, show major abnormalities in the cytoarchitecture of the neocortex and cerebellum and die within 24 h of birth. Later-born neurons do not localize properly in the cortex. A similar mislocalization is observed with cerebellar Purkinje neurons. Although the proportion of astrocytes is higher than normal, the numbers of oligodendrocytes are reduced. In contrast, conditional knockout of HDAC3 in neurons of the forebrain and certain other brain regions, using Thy1-Cre and calcium/calmodulin dependent protein kinase II α-Cre for ablation, produces no overt abnormalities in the organization of cells within the cortex or of cerebellar Purkinje neurons at birth. However, both lines of conditional knockout mice suffer from progressive hind limb paralysis and ataxia and die around 6 weeks after birth. The mice display an increase in overall numbers of cells, higher numbers of astrocytes, and Purkinje neuron degeneration. Taken together, our results demonstrate that HDAC3 plays an essential role in regulating brain development, with effects on both neurons and glia in different brain regions. PMID:25339172
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Suzuki, S; Nakamura, S; Sakaguchi, T; Mitsuoka, H; Tsuchiya, Y; Kojima, Y; Konno, H; Baba, S
1998-11-01
Animal models of total hepatic ischemia (THI) and reperfusion injury are restricted by concomitant splanchnic congestion. This study was performed to determine the requirement suitable for an extracorporeal portosystemic shunt (PSS) to maintain the intestinal integrity in a rat model of THI. Using a polyethylene tube (0.86 or 1 mm i.d.), PSS was placed between the mesenteric and jugular veins. Comparison was done between THI models with or without PSS and a partial ischemia model with hepatectomy of the nonischemic lobes. Well-tolerated hepatic ischemic period, portal pressure after 10 min of hepatic ischemia, portal endotoxin levels at 1 h after reperfusion, histological features of the small bowel just before reperfusion, and local jejunal and ileal blood hemoglobin oxygen saturation index (ISO2) were compared among the models. Animals without PSS poorly tolerated 30 min of THI. Animals receiving THI with PSS or partial hepatic ischemia tolerated a longer ischemic period (60 min) with a significantly higher small bowel ISO2, lower portal pressure and endotoxin levels (P < 0.01), and less histological damage of the small bowel when compared to those receiving THI without PSS. Portal endotoxin levels after THI with PSS using a 1-mm i.d. tube as well as partial hepatic ischemia were significantly lower than those after THI with PSS using a 0.86-mm i.d. tube. THI with PSS using a 1-mm i.d. tube was strikingly similar to partial hepatic ischemia in the pathophysiological profile during hepatic ischemia. PSS with a tube 1 mm or more in inner diameter offers pathophysiological advantages in experiments on THI and reperfusion. Copyright 1998 Academic Press.
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Abeta42-driven cerebral amyloidosis in transgenic mice reveals early and robust pathology.
Radde, Rebecca; Bolmont, Tristan; Kaeser, Stephan A; Coomaraswamy, Janaky; Lindau, Dennis; Stoltze, Lars; Calhoun, Michael E; Jäggi, Fabienne; Wolburg, Hartwig; Gengler, Simon; Haass, Christian; Ghetti, Bernardino; Czech, Christian; Hölscher, Christian; Mathews, Paul M; Jucker, Mathias
2006-09-01
We have generated a novel transgenic mouse model on a C57BL/6J genetic background that coexpresses KM670/671NL mutated amyloid precursor protein and L166P mutated presenilin 1 under the control of a neuron-specific Thy1 promoter element (APPPS1 mice). Cerebral amyloidosis starts at 6-8 weeks and the ratio of human amyloid (A)beta42 to Abeta40 is 1.5 and 5 in pre-depositing and amyloid-depositing mice, respectively. Consistent with this ratio, extensive congophilic parenchymal amyloid but minimal amyloid angiopathy is observed. Amyloid-associated pathologies include dystrophic synaptic boutons, hyperphosphorylated tau-positive neuritic structures and robust gliosis, with neocortical microglia number increasing threefold from 1 to 8 months of age. Global neocortical neuron loss is not apparent up to 8 months of age, but local neuron loss in the dentate gyrus is observed. Because of the early onset of amyloid lesions, the defined genetic background of the model and the facile breeding characteristics, APPPS1 mice are well suited for studying therapeutic strategies and the pathomechanism of amyloidosis by cross-breeding to other genetically engineered mouse models.
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Aβ42-driven cerebral amyloidosis in transgenic mice reveals early and robust pathology
Radde, Rebecca; Bolmont, Tristan; Kaeser, Stephan A; Coomaraswamy, Janaky; Lindau, Dennis; Stoltze, Lars; Calhoun, Michael E; Jäggi, Fabienne; Wolburg, Hartwig; Gengler, Simon; Haass, Christian; Ghetti, Bernardino; Czech, Christian; Hölscher, Christian; Mathews, Paul M; Jucker, Mathias
2006-01-01
We have generated a novel transgenic mouse model on a C57BL/6J genetic background that coexpresses KM670/671NL mutated amyloid precursor protein and L166P mutated presenilin 1 under the control of a neuron-specific Thy1 promoter element (APPPS1 mice). Cerebral amyloidosis starts at 6–8 weeks and the ratio of human amyloid (A)β42 to Aβ40 is 1.5 and 5 in pre-depositing and amyloid-depositing mice, respectively. Consistent with this ratio, extensive congophilic parenchymal amyloid but minimal amyloid angiopathy is observed. Amyloid-associated pathologies include dystrophic synaptic boutons, hyperphosphorylated tau-positive neuritic structures and robust gliosis, with neocortical microglia number increasing threefold from 1 to 8 months of age. Global neocortical neuron loss is not apparent up to 8 months of age, but local neuron loss in the dentate gyrus is observed. Because of the early onset of amyloid lesions, the defined genetic background of the model and the facile breeding characteristics, APPPS1 mice are well suited for studying therapeutic strategies and the pathomechanism of amyloidosis by cross-breeding to other genetically engineered mouse models. PMID:16906128
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Changes in morphology of retinal ganglion cells with eccentricity in retinal degeneration.
Anderson, E E; Greferath, U; Fletcher, E L
2016-05-01
Ganglion cells are the output neurons of the retina and are known to remodel during the subtle plasticity changes that occur following the death of photoreceptors in inherited retinal degeneration. We examine the influence of retinal eccentricity on anatomical remodelling and ganglion cell morphology well after photoreceptor loss. Rd1 mice that have a mutation in the β subunit of phosphodiesterase 6 were used as a model of retinal degeneration and gross remodelling events were examined by processing serial sections for immunocytochemistry. Retinal wholemounts from rd1-Thy1 and control Thy1 mice that contained a fluorescent protein labelling a subset of ganglion cells were processed for immunohistochemistry at 11 months of age. Ganglion cells were classified based on their soma size, dendritic field size and dendritic branching pattern and their dendritic fields were analysed for their length, area and quantity of branching points. Overall, more remodelling was found in the central compared with the peripheral retina. In addition, the size and complexity of A2, B1, C1 and D type ganglion cells located in the central region of the retina decreased. We propose that the changes in ganglion cell morphology are correlated with remodelling events in these regions and impact the function of retinal circuitry in the degenerated retina.
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Friedmacher, Florian; Hofmann, Alejandro Daniel; Takahashi, Hiromizu; Takahashi, Toshiaki; Gosemann, Jan-Hendrik; Puri, Prem
2014-02-01
Pulmonary hypoplasia (PH), characterized by alveolar immaturity, remains the main cause of neonatal mortality and long-term morbidity in infants with congenital diaphragmatic hernia (CDH). Lipid-containing interstitial fibroblasts (LIFs) are critically important for normal alveolar development. Thymocyte antigen 1 (Thy-1) is a highly expressed cell-surface protein in this specific subset of lung fibroblasts, which plays a key role in fetal alveolarization by coordinating the differentiation and lipid homeostasis of alveolar LIFs. Thy-1 increases the lipid content of LIFs by upregulation of adipocyte differentiation-related protein (ADRP), a lipogenic molecular marker characterizing pulmonary LIFs. Thy-1 (-/-) mice further show impaired alveolar development with reduced proliferation of pulmonary LIFs, resulting in a PH-similar phenotype. We hypothesized that pulmonary Thy-1 signaling is disrupted in experimentally induced CDH, which may has an adverse effect on the lipid content of alveolar LIFs. Timed-pregnant Sprague-Dawley rats were treated with either 100 mg nitrofen or vehicle on embryonic day 9.5 (E9.5). Fetuses were killed on E21.5, and lungs were divided into controls (n = 14) and CDH-associated PH (n = 14). Pulmonary gene expression levels of Thy-1 and ADRP were assessed by quantitative real-time PCR. ADRP immunohistochemistry and oil-red-O staining were used to localize alveolar LIF expression and lipid droplets. Immunofluorescence double staining for Thy-1 and oil-red-O was performed to evaluate Thy-1 expression and lipid content in alveolar LIFs. Radial alveolar count was significantly reduced in CDH-associated PH with significant downregulation of pulmonary Thy-1 and ADRP mRNA expression compared to controls. ADRP immunoreactivity and lipid droplets were markedly diminished in alveolar interstitial cells, which coincided with decreased alveolar LIF expression in CDH-associated PH compared to controls. Confocal laser scanning microscopy confirmed markedly decreased Thy-1 expression and lipid content in alveolar LIFs of CDH-associated PH compared to controls. Our study provides strong evidence that disruption of pulmonary Thy-1 signaling results in reduced lipid droplets in alveolar LIFs and may thus contribute to PH in the nitrofen-induced CDH model. Treatment modalities aimed at increasing lipid content in alveolar LIFs may therefore have a therapeutic potential in attenuating CDH-associated PH.
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Exploring the brain on multiple scales with correlative two-photon and light sheet microscopy
NASA Astrophysics Data System (ADS)
Silvestri, Ludovico; Allegra Mascaro, Anna Letizia; Costantini, Irene; Sacconi, Leonardo; Pavone, Francesco S.
2014-02-01
One of the unique features of the brain is that its activity cannot be framed in a single spatio-temporal scale, but rather spans many orders of magnitude both in space and time. A single imaging technique can reveal only a small part of this complex machinery. To obtain a more comprehensive view of brain functionality, complementary approaches should be combined into a correlative framework. Here, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, taking advantage of blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living thy1-GFP-M mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from the apical portion, the whole pyramidal neuron can then be segmented. The correlative approach presented here allows contextualizing within a three-dimensional anatomic framework the neurons whose dynamics have been observed with high detail in vivo.
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Podocyte changes upon induction of albuminuria in Thy-1.1 transgenic mice.
Smeets, Bart; Dijkman, Henry B P M; te Loeke, Nathalie A J M; van Son, Jacco P H F; Steenbergen, Eric J; Assmann, Karel J M; Wetzels, Jack F M; Groenen, Patricia J T A
2003-12-01
Thy-1.1 transgenic mice, characterized by ectopic expression of the Thy-1.1 protein on podocytes, spontaneously develop proteinuria and focal glomerulosclerosis (FGS). Injection of a monoclonal antibody (mAb) directed against the Thy-1.1 protein in young transgenic mice induces a massive albuminuria that is followed by an accelerated FGS within 3 weeks. This albuminuria is complement and leukocyte independent. The time course of proteinuria, the pathogenesis of the acute proteinuria and the dose dependency of FGS are unknown. Albuminuria was measured in Thy-1.1 transgenic mice after injection of different doses of anti-Thy-1.1 mAb and at different time points within the first 24 h after injection. Podocytic foot processes and slit pore diameter were quantitated by electron microscopy. Changes in expression of slit pore constituents (podocin, CD2AP, nephrin and ZO-1), cytoskeleton-associated proteins (actin, alpha-actinin, ezrin and synaptopodin), the GDH-podocyte adhesion molecules alpha(3)-integrin, and heparan sulfate were studied by immunofluorescence. FGS was scored by light microscopy at 3 weeks after induction of albuminuria. Albuminuria in Thy-1.1 transgenic mice was observed within 10 min after anti-Thy-1.1 mAb injection. This rapid development of albuminuria was accompanied by a reduction in number of podocytic foot processes from 20.0 +/- 0.7/10 microm glomerular basement membrane (GBM) in saline-treated transgenic mice to 8.0 +/- 0.5 and 2.2 +/- 0.2 in anti-Thy-1.1-treated mice, at 10 min and 8 h after treatment, respectively. In addition, we observed a significant decrease in width of remaining slit pores, from 32.7 +/- 1.1 to 26.8 +/- 1.4 nm at 10 min after mAb injection. By immunofluorescence, we did not observe major changes in the expression pattern of any of the proteins studied. There was no correlation between the injected dose of the anti-Thy-1.1 mAb and the acute albuminuria. In contrast, the percentage of FGS at 3 weeks correlated with the dose, and a significant correlation between the percentage of FGS and the time-averaged albuminuria over the 3 week study period (P < 0.001) was found. Injection of mAb directed against the Thy-1.1 protein, in young non-albuminuric Thy-1.1 transgenic mice, induced an acute albuminuria within 10 min, which was accompanied by foot process effacement. Notably, we observed a decrease in slit pore width although the expression of slit pore proteins was unchanged. Also, the acute albuminuria could not be related to alterations in cytoskeleton-associated proteins, the GBM adhesion molecule alpha(3)-integrin or heparan sulfate in the GBM. The dose-dependent development of FGS and the correlation between the percentage FGS and time-averaged albuminuria suggest that, in our model, FGS is a consequence of podocyte injury. However, the data leave open the possibility that albuminuria itself contributes to FGS development. The Thy-1.1 transgenic mouse model is an excellent model to study further the relationship between podocytic injury, albuminuria and the development of FGS.
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Aid, Saba; Parikh, Nishant; Palumbo, Sara; Bosetti, Francesca
2010-07-12
Neuroinflammation is a critical component in the progression of several neurological and neurodegenerative diseases and cyclooxygenases (COX)-1 and -2 are key regulators of innate immune responses. We recently demonstrated that COX-1 deletion attenuates, whereas COX-2 deletion enhances, the neuroinflammatory response, blood-brain barrier permeability and leukocyte recruitment during lipopolysaccharide (LPS)-induced innate immune activation. Here, we used transgenic mice, which overexpressed human COX-2 via neuron-specific Thy-1 promoter (TgCOX-2), causing elevated prostaglandins (PGs) levels. We tested whether neuronal COX-2 overexpression affects the glial response to a single intracerebroventricular injection of LPS, which produces a robust neuroinflammatory reaction. Relative to non-transgenic controls (NTg), 7 month-old TgCOX-2 did not show any basal neuroinflammation, as assessed by gene expression of markers of inflammation and oxidative stress, neuronal damage, as assessed by Fluoro-JadeB staining, or systemic inflammation, as assessed by plasma levels of IL-1beta and corticosterone. Twenty-four hours after LPS injection, all mice showed increased microglial activation, as indicated by Iba1 immunostaining, neuronal damage, mRNA expression of cytokines (TNF-alpha, IL-6), reactive oxygen expressing enzymes (iNOS and NADPH oxidase subunits), endogenous COX-2, cPLA(2) and mPGES-1, and hippocampal and cortical IL-1beta levels. However, the increases were similar in TgCOX-2 and NTg. In NTg, LPS increased brain PGE(2) to the levels observed in TgCOX-2. These results suggest that PGs derived from neuronal COX-2 do not play a role in the neuroinflammatory response to acute activation of brain innate immunity. This is likely due to the direct effect of LPS on glial rather than neuronal cells. Published by Elsevier Ireland Ltd.
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Characterization of cell types during rat liver development.
Fiegel, Henning C; Park, Jonas J h; Lioznov, Michael V; Martin, Andreas; Jaeschke-Melli, Stefan; Kaufmann, Peter M; Fehse, Boris; Zander, Axel R; Kluth, Dietrich
2003-01-01
Hepatic stem cells have been identified in adult liver. Recently, the origin of hepatic progenitors and hepatocytes from bone marrow was demonstrated. Hematopoietic and hepatic stem cells share the markers CD 34, c-kit, and Thy1. Little is known about liver stem cells during liver development. In this study, we investigated the potential stem cell marker Thy1 and hepatocytic marker CK-18 during liver development to identify putative fetal liver stem cell candidates. Livers were harvested from embryonic and fetal day (ED) 16, ED 18, ED 20, and neonatal ED 22 stage rat fetuses from Sprague-Dawley rats. Fetal livers were digested by collagenase-DNAse solution and purified by percoll centrifugation. Magnetic cell sorting (MACS) depletion of fetal liver cells was performed using OX43 and OX44 antibodies. Cells were characterized by immunocytochemistry for Thy1, CK-18, and proliferating cell antigen Ki-67 and double labeling for Thy1 and CK-18. Thy1 expression was found at all stages of liver development before and after MACS in immunocytochemistry. Thy1 positive cells were enriched after MACS only in early developmental stages. An enrichment of CK-18 positive cells was found after MACS at all developmental stages. Cells coexpressing Thy1 and CK-18 were identified by double labeling of fetal liver cell isolates. In conclusion, hepatic progenitor cells (CK-18 positive) in fetal rat liver express Thy1. Other progenitors express only CK-18. This indicates the coexistence of different hepatic cell compartments. Isolation and further characterization of such cells is needed to demonstrate their biologic properties.
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Henríquez, Mauricio; Herrera-Molina, Rodrigo; Valdivia, Alejandra; Alvarez, Alvaro; Kong, Milene; Muñoz, Nicolás; Eisner, Verónica; Jaimovich, Enrique; Schneider, Pascal; Quest, Andrew F. G.; Leyton, Lisette
2011-01-01
Thy-1, an abundant mammalian glycoprotein, interacts with αvβ3 integrin and syndecan-4 in astrocytes and thus triggers signaling events that involve RhoA and its effector p160ROCK, thereby increasing astrocyte adhesion to the extracellular matrix. The signaling cascade includes calcium-dependent activation of protein kinase Cα upstream of Rho; however, what causes the intracellular calcium transients required to promote adhesion remains unclear. Purinergic P2X7 receptors are important for astrocyte function and form large non-selective cation pores upon binding to their ligand, ATP. Thus, we evaluated whether the intracellular calcium required for Thy-1-induced cell adhesion stems from influx mediated by ATP-activated P2X7 receptors. Results show that adhesion induced by the fusion protein Thy-1-Fc was preceded by both ATP release and sustained intracellular calcium elevation. Elimination of extracellular ATP with Apyrase, chelation of extracellular calcium with EGTA, or inhibition of P2X7 with oxidized ATP, all individually blocked intracellular calcium increase and Thy-1-stimulated adhesion. Moreover, Thy-1 mutated in the integrin-binding site did not trigger ATP release, and silencing of P2X7 with specific siRNA blocked Thy-1-induced adhesion. This study is the first to demonstrate a functional link between αvβ3 integrin and P2X7 receptors, and to reveal an important, hitherto unanticipated, role for P2X7 in calcium-dependent signaling required for Thy-1-stimulated astrocyte adhesion. PMID:21502139
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Optogenetic noise-photostimulation on the brain increases somatosensory spike firing responses.
Huidobro, Nayeli; De la Torre-Valdovinos, Braniff; Mendez, Abraham; Treviño, Mario; Arias-Carrion, Oscar; Chavez, Fermin; Gutierrez, Ranier; Manjarrez, Elias
2018-01-18
We examined whether the optogenetic noise-photostimulation (ONP) of the barrel cortex (BC) of anesthetized Thy1-ChR2-YFP transgenic mice increases the neuronal multiunit-activity response evoked by whisker mechanical stimulation (whisker-evoked MUA). In all transgenic mice, we found that the signal-to-noise ratio (SNR) of such whisker-evoked MUA signals exhibited an inverted U-like shape as a function of the ONP level. Numerical simulations of a ChR2-expressing neuron model qualitatively support our experimental data. These results show that the application of an intermediate intensity of ONP in the brain can increase cortical somatosensory spike responses to whisker protraction. These findings suggest that ONP of the mice-BC could produce improvements in somatosensory perception to whisker stimulation. Copyright © 2017 Elsevier B.V. All rights reserved.
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Cánovas, José; Berndt, F Andrés; Sepúlveda, Hugo; Aguilar, Rodrigo; Veloso, Felipe A; Montecino, Martín; Oliva, Carlos; Maass, Juan C; Sierralta, Jimena; Kukuljan, Manuel
2015-05-13
The acquisition of distinct neuronal fates is fundamental for the function of the cerebral cortex. We find that the development of subcerebral projections from layer 5 neurons in the mouse neocortex depends on the high levels of expression of the transcription factor CTIP1; CTIP1 is coexpressed with CTIP2 in neurons that project to subcerebral targets and with SATB2 in those that project to the contralateral cortex. CTIP1 directly represses Tbr1 in layer 5, which appears as a critical step for the acquisition of the subcerebral fate. In contrast, lower levels of CTIP1 in layer 6 are required for TBR1 expression, which directs the corticothalamic fate. CTIP1 does not appear to play a critical role in the acquisition of the callosal projection fate in layer 5. These findings unravel a key step in the acquisition of cell fate for closely related corticofugal neurons and indicate that differential dosages of transcriptions factors are critical to specify different neuronal identities. Copyright © 2015 the authors 0270-6474/15/357552-13$15.00/0.
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Cylindromatosis mediates neuronal cell death in vitro and in vivo.
Ganjam, Goutham K; Terpolilli, Nicole Angela; Diemert, Sebastian; Eisenbach, Ina; Hoffmann, Lena; Reuther, Christina; Herden, Christiane; Roth, Joachim; Plesnila, Nikolaus; Culmsee, Carsten
2018-01-19
The tumor-suppressor cylindromatosis (CYLD) is a deubiquitinating enzyme and key regulator of cell proliferation and inflammation. A genome-wide siRNA screen linked CYLD to receptor interacting protein-1 (RIP1) kinase-mediated necroptosis; however, the exact mechanisms of CYLD-mediated cell death remain unknown. Therefore, we investigated the precise role of CYLD in models of neuronal cell death in vitro and evaluated whether CYLD deletion affects brain injury in vivo. In vitro, downregulation of CYLD increased RIP1 ubiquitination, prevented RIP1/RIP3 complex formation, and protected neuronal cells from oxidative death. Similar protective effects were achieved by siRNA silencing of RIP1 or RIP3 or by pharmacological inhibition of RIP1 with necrostatin-1. In vivo, CYLD knockout mice were protected from trauma-induced brain damage compared to wild-type littermate controls. These findings unravel the mechanisms of CYLD-mediated cell death signaling in damaged neurons in vitro and suggest a cell death-mediating role of CYLD in vivo.
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Nasr, Mohammed A F
2017-08-01
The current study was aimed to investigate the impact of THI on productive and reproductive indices of PE and F 1 crosses (50% PE and 50% Italian buffaloes) and back crosses (BC) (75% PE and 25% Italian buffaloes) under hot conditions. In this study, 8385 records used PE (1914, 1518, and 1737), F 1 (387, 447, and 657), and BC (495, 585, and 645) for low, medium, and high THI, respectively. The high THI reduced the conception rate after first insemination in PE, F 1 , and BC ((odds ratio, OR) = 1.187, P = 0.007; 2.361, <0.0001 and 1.603, <0.0001, respectively) when compared with low THI. But, stillbirth and calving condition were not significantly influenced by different THI levels in BC and F 1 . BC was more bearable to the harsh environment; they possessed the highest incidence of conception after first insemination (72.70, 72.60, and 62.40%), producing live calves (98.50, 100, and 99.40%) with easy calving condition (98.50, 100, and 99.40%) when compared to PE and F 1 at different levels of THI, respectively. The average daily milk yield and peak of milk production were decreased in PE (4.02 and 5.12%), BC (13.33 and 10.95%), and F 1 (25.29 and 12.20%) from low to high THI. However, BC revealed no significant changes in days open, dry period, calving interval, gestation length, service per conception, and the first service post-partum at different levels of THI when compared with PE and F 1 . Therefore, rearing BC is recommended for improving buffalo productive and reproductive performance under hot conditions.
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Thi Qar Bee Farm Thi Qar, Iraq
2010-04-01
vegetation and fields where bees once gathered pollen and beekeepers face hardships from droughts and lack of financial assistance. 1...of equipment, and provided training to the bee farmers. General topography of the area was flat with vacant or agricultural land extending for a...OFFICE OF THE SPECIAL INSPECTOR GENERAL FOR IRAQ RECONSTRUCTION THI QAR BEE FARM THI QAR, IRAQ SIGIR PA--09--188
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Schüller, L K; Burfeind, O; Heuwieser, W
2014-05-01
The objectives of this retrospective study were to investigate the relationship between temperature-humidity index (THI) and conception rate (CR) of lactating dairy cows, to estimate a threshold for this relationship, and to identify periods of exposure to heat stress relative to breeding in an area of moderate climate. In addition, we compared three different heat load indices related to CR: mean THI, maximum THI, and number of hours above the mean THI threshold. The THI threshold for the influence of heat stress on CR was 73. It was statistically chosen based on the observed relationship between the mean THI at the day of breeding and the resulting CR. Negative effects of heat stress, however, were already apparent at lower levels of THI, and 1 hour of mean THI of 73 or more decreased the CR significantly. The CR of lactating dairy cows was negatively affected by heat stress both before and after the day of breeding. The greatest negative impact of heat stress on CR was observed 21 to 1 day before breeding. When the mean THI was 73 or more in this period, CR decreased from 31% to 12%. Compared with the average maximum THI and the total number of hours above a threshold of more than or 9 hours, the mean THI was the most sensitive heat load index relating to CR. These results indicate that the CR of dairy cows raised in the moderate climates is highly affected by heat stress. Copyright © 2014 Elsevier Inc. All rights reserved.
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Intimate association of Thy-1 and the T-cell antigen receptor with the CD45 tyrosine phosphatase
DOE Office of Scientific and Technical Information (OSTI.GOV)
Volarevic, S.; Burns, C.M.; Sussman, J.J.
1990-09-01
Immunoprecipitation of Thy-1 from Triton X-100 detergent lysates of surface-iodinated and chemically cross-linked T cells precipitated at least first major and discrete bands. Four of these bands were identified as Thy-1, CD45 (a trasmembrane tyrosine phosphatase), a major histocompatibility complex-encoded class I molecule, and {beta}{sub 2}-microglobulin. Similar analyses revealed that CD45 was coprecipitated from lysates of cross-linker-treated cells by antibodies to the T-cell antigen receptor (TCR). The same pattern of coprecipitated bands was observed when digitonin was used to lyse untreated cells. Immunoprecipitation of Thy-1 or the TCR from lysates of cross-linked T cells precipitated CD45 tyrosine phosphatase activity. Calculationsmore » based upon the amounts of coprecipitated enzymatic activity or TCR {zeta} chain indicate that a substantial fraction of Thy-1 and TCR complexes can be cross-linked to CD45. These data support a model in which the dependence of Thy-1 signaling on TCR coexpression is due to their common interaction with a tyrosine phosphatase and provide a possible structural basis for the influence of CD45 on TCR-mediated signaling.« less
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[Preliminary investigation of psychologic factors in 76 tinnitus patients].
Mao, Kunhua; Jiang, Wen; Feng, Yong
2011-08-01
To study the psychological aspects of tinnitus patients, to analyze the distribution of psychologic obstacle in tinnitus patients, and then to provide information for diagnosing and treating tinnitus clinically. All patients were detected their frequency and loudness of tinnitus. Then they were evaluated by symptom checklist 90 (SCL-90), life satisfaction scale, Pittsburgh sleep quality index (PSQI) and tinnitus handicap inventory (THI). All data were analyzed with statistical software SPSS11.0. (1)There was no straight line correlation between frequency, loudness of tinnitus and the patient's scores from SCL-90, life satisfaction rating scale (LSR), life satisfaction index A (LSIA), LSIB, PSQI, THI. (2) To 76 tinnitus patients, some factors of SCL-90 were higher than internal nom. Compared with internal nom, tinnitus patients' score of LSR, LSIA and LSIB were all lower than it. Many of tinnitus patients had sleep disorder, the ratio was higher than internal nom. (3) Grouping these patients, based on the score of THI. To THI four grade group and THI five grade group, their satisfaction of lives were lower, some factors of SCL-90 were higher than internal nom. To THI five grade group, the ratio about sleep disorder was higher than internal nom. There is no straight line correlation between frequency, loudness of tinnitus and the patient's scores from SCL-90, LSR, LSIA, LSIB, PSQ1, THI. Grouping based on the score of THI, the groups of THI four grade and THI five grade are approved that they have psychologic obstacle obviously, they should be paid close attention.
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Mutsaers, Chantal A.; Thomson, Derek; Hamilton, Gillian; Parson, Simon H.; Gillingwater, Thomas H.
2012-01-01
Spinal muscular atrophy (SMA) is a leading genetic cause of infant mortality, resulting primarily from the degeneration and loss of lower motor neurons. Studies using mouse models of SMA have revealed widespread heterogeneity in the susceptibility of individual motor neurons to neurodegeneration, but the underlying reasons remain unclear. Data from related motor neuron diseases, such as amyotrophic lateral sclerosis (ALS), suggest that morphological properties of motor neurons may regulate susceptibility: in ALS larger motor units innervating fast-twitch muscles degenerate first. We therefore set out to determine whether intrinsic morphological characteristics of motor neurons influenced their relative vulnerability to SMA. Motor neuron vulnerability was mapped across 10 muscle groups in SMA mice. Neither the position of the muscle in the body, nor the fibre type of the muscle innervated, influenced susceptibility. Morphological properties of vulnerable and disease-resistant motor neurons were then determined from single motor units reconstructed in Thy.1-YFP-H mice. None of the parameters we investigated in healthy young adult mice – including motor unit size, motor unit arbor length, branching patterns, motor endplate size, developmental pruning and numbers of terminal Schwann cells at neuromuscular junctions - correlated with vulnerability. We conclude that morphological characteristics of motor neurons are not a major determinant of disease-susceptibility in SMA, in stark contrast to related forms of motor neuron disease such as ALS. This suggests that subtle molecular differences between motor neurons, or extrinsic factors arising from other cell types, are more likely to determine relative susceptibility in SMA. PMID:23285108
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Ozaki, Mitsunori; Sano, Hiromi; Sato, Shigeki; Ogura, Mitsuhiro; Mushiake, Hajime; Chiken, Satomi; Nakao, Naoyuki; Nambu, Atsushi
2017-12-01
To understand how information from different cortical areas is integrated and processed through the cortico-basal ganglia pathways, we used optogenetics to systematically stimulate the sensorimotor cortex and examined basal ganglia activity. We utilized Thy1-ChR2-YFP transgenic mice, in which channelrhodopsin 2 is robustly expressed in layer V pyramidal neurons. We applied light spots to the sensorimotor cortex in a grid pattern and examined neuronal responses in the globus pallidus (GP) and entopeduncular nucleus (EPN), which are the relay and output nuclei of the basal ganglia, respectively. Light stimulation typically induced a triphasic response composed of early excitation, inhibition, and late excitation in GP/EPN neurons. Other response patterns lacking 1 or 2 of the components were also observed. The distribution of the cortical sites whose stimulation induced a triphasic response was confined, whereas stimulation of the large surrounding areas induced early and late excitation without inhibition. Our results suggest that cortical inputs to the GP/EPN are organized in a "local inhibitory and global excitatory" manner. Such organization seems to be the neuronal basis for information processing through the cortico-basal ganglia pathways, that is, releasing and terminating necessary information at an appropriate timing, while simultaneously suppressing other unnecessary information. © The Author 2017. Published by Oxford University Press.
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Phenotypic, molecular, and functional characterization of human peripheral blood CD34+/THY1+ cells.
Humeau, L; Bardin, F; Maroc, C; Alario, T; Galindo, R; Mannoni, P; Chabannon, C
1996-02-01
A subset of mobilized CD34+ cells present in patient aphereses expresses Thy1 (CDw90). This population contains most long-term culture initiating cells, as assayed with a murine stromal cell line. It also contains a significant proportion of colony-forming unit granulocyte macrophage, but very few burst-forming unit erythroid. The limited differentiation towards the erythroid lineage is further confirmed by the absence of GATA-1 mRNA in the CD34+/Thy1+ subset, and by the low level of c-kit expression. The CD34+/Thy1+ subset appears phenotypically and functionally heterogeneous, a finding consistent with its high representation, compared to phenotypes such as CD34+/CD38-. Therefore, while at least some of CD34+/Thy1+ cells may be infectable by retroviral vectors, as shown by the presence of a transcript for the receptor for murine amphotropic retroviruses, the use of this selection strategy to specifically target human stem cells appears questionable.
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Wang, Kuifeng; Wang, Qi; Chen, Jing; Chen, Lili; Jiang, Hualiang; Shen, Xu
2011-01-01
Thymidylate synthase X (ThyX) catalyzes the methylation of dUMP to form dTMP in bacterial life cycle and is regarded as a promising target for antibiotics discovery. Helicobacter pylori is a human pathogen associated with a number of human diseases. Here, we cloned and purified the ThyX enzyme from H. pylori SS1 strain (HpThyX). The recombinant HpThyX was discovered to exhibit the maximum activity at pH 8.5, and Km values of the two substrates dUMP and CH2H4folate were determined to be 15.3 ± 1.25 μM and 0.35 ± 0.18 mM, respectively. The analyzed crystal structure of HpThyX with the cofactor FAD and the substrate dUMP (at 2.31 Å) revealed that the enzyme was a tetramer bound to four dUMP and four FAD molecules. Different from the catalytic feature of the classical thymidylate synthase (ThyA), N5 atom of the FAD functioned as a nucleophile in the catalytic reaction instead of Ser84 and Ser85 residues. Our current work is expected to help better understand the structural and enzymatic features of HpThyX thus further providing valuable information for anti-H. pylori inhibitor discovery. PMID:21633987
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Zhou, Wen-Qin; Wang, Peng; Shao, Qiu-Ping; Wang, Jian
2016-08-01
Acute respiratory distress syndrome (ARDS) is a common clinical disorder characterized by pulmonary edema leading to acute lung damage and arterial hypoxemia. Pulmonary fibrosis is a progressive, fibrotic lung disorder, whose pathogenesis in ARDS remains speculative. LincRNA-p21 was a novel regulator of cell proliferation, apoptosis and DNA damage response. This study aims to investigate the effects and mechanism of lincRNA-p21 on pulmonary fibrosis in ARDS. Purified 10 mg/kg LPS was dropped into airways of C57BL/6 mice. Expression levels of lincRNA-p21 and Thy-1 were measured by real-time PCR or western blotting. Proliferation of lung fibroblasts was analyzed by BrdU incorporation assay. Lung and BAL collagen contents were estimated using colorimetric Sircol assay. LincRNA-p21 expression was time-dependently increased and Thy-1 expression was time-dependently reduced in a mouse model of ARDS and in LPS-treated lung fibroblasts. Meanwhile, lung fibroblast proliferation was also time-dependently elevated in LPS-treated lung fibroblasts. In addition, lung fibroblast proliferation could be promoted by lincRNA-p21 overexpression and LPS treatment, however, the elevated lung fibroblast proliferation was further abrogated by Thy-1 overexpression or lincRNA-p21 interference. And Thy-1 interference could elevate cell viability of lung fibroblasts and rescue the reduction of lung fibroblast proliferation induced by lincRNA-p21 interference. Moreover, lincRNA-p21 overexpression dramatically inhibited acetylation of H3 and H4 at the Thy-1 promoter and Thy-1 expression levels in HLF1 cells. Finally, lincRNA-p21 interference rescued LPS-induced increase of lung and BAL collagen contents. LincRNA-p21 could lead to pulmonary fibrosis in ARDS by inhibition of the expression of Thy-1.
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Nagae, Miwa; Parniske, Martin; Kawaguchi, Masayoshi; Takeda, Naoya
2016-12-01
Lotus japonicus THIC is expressed in all organs, and the encoded protein catalyzes thiamine biosynthesis. Loss of function produces chlorosis, a typical thiamine-deficiency phenotype, and mortality. To investigate thiamine's role in symbiosis, we focused on THI1, a thiamine-biosynthesis gene expressed in roots, nodules, and seeds. The thi1 mutant had green leaves, but formed small nodules and immature seeds. These phenotypes were rescued by THI1 complementation and by exogenous thiamine. Thus, THI1 is required for nodule enlargement and seed maturation. On the other hand, colonization by arbuscular mycorrhiza (AM) fungus Rhizophagus irregularis was not affected in the thi1 mutant or by exogenous thiamine. However, spores of R. irregularis stored more thiamine than the source (host plants), despite lacking thiamine biosynthesis genes. Therefore, disturbance of the thiamine supply would affect progeny phenotypes such as spore formation and hyphal growth. Further investigation will be required to elucidate thiamine's effect on AM.
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Schüller, Laura K; Heuwieser, Wolfgang
2016-08-01
The objectives of this study were to examine heat stress conditions at cow level and to investigate the relationship to the climate conditions at 5 different stationary locations inside a dairy barn. In addition, we compared the climate conditions at cow level between primiparous and multiparous cows for a period of 1 week after regrouping. The temperature-humidity index (THI) differed significantly between all stationary loggers. The lowest THI was measured at the window logger in the experimental stall and the highest THI was measured at the central logger in the experimental stall. The THI at the mobile cow loggers was 2·33 THI points higher than at the stationary loggers. Furthermore, the mean daily THI was higher at the mobile cow loggers than at the stationary loggers on all experimental days. The THI in the experimental pen was 0·44 THI points lower when the experimental cow group was located inside the milking parlour. The THI measured at the mobile cow loggers was 1·63 THI points higher when the experimental cow group was located inside the milking parlour. However, there was no significant difference for all climate variables between primiparous and multiparous cows. These results indicate, there is a wide range of climate conditions inside a dairy barn and especially areas with a great distance to a fresh air supply have an increased risk for the occurrence of heat stress conditions. Furthermore, the heat stress conditions are even higher at cow level and cows not only influence their climatic environment, but also generate microclimates within different locations inside the barn. Therefore climate conditions should be obtained at cow level to evaluate the heat stress conditions that dairy cows are actually exposed to.
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The role of water in generating Fe-depletion and the calc-alkaline trend
NASA Astrophysics Data System (ADS)
Zimmer, M. M.; Plank, T.
2006-12-01
Describing a magmatic suite as calc-alkaline (CA) or tholeiitic (TH) is a first order characterization, but existing classification schemes (AFM ternary plots and FeO*/MgO vs. SiO2) may convolute magmatic processes and can result in contradictory classification. The salient feature of TH vs. CA evolution is the extent of Fe enrichment or depletion in the magma. A plot of FeO* vs. MgO provides the most straightforward way to quantify Fe enrichment and to develop models for its origin. We present a new quantitative classification utilizing the FeO*-MgO plot, the tholeiitic index (THI) = Fe3-5/Fe8 (Fe3-5=average FeO* at 3-5 wt% MgO; Fe8=FeO* at 8 wt% MgO). THI of 1.2 indicates 20% FeO* enrichment from a magma's starting composition at Fe8, while THI of 0.8 indicates 20% depletion in FeO*. A magmatic suite is CA if THI is <1, and TH if THI is >1. Arcs range from 0.6 to 1.1, back arc basins from 1.1-1.3, and MORBs are \\ge1.6. This classification allows comparison of magmatic evolution on a global basis, regardless of starting composition, and is useful for quantitative comparison to liquid line of descent models. Hypotheses for generating CA magmas include high water contents, high pressure of crystallization, high oxygen fugacity, and high Mg# andesitic starting compositions. In order to test the control of H2O, we compare the THI to average magmatic water contents from undegassed melt inclusions and glasses (S>1000 ppm or CO2>50 ppm) from twenty-eight arc volcanoes and back arc basins, including new water contents from seven Aleutian volcanoes. The resulting negative correlation (R2=0.8) between water concentration and THI (with end-members at 0.8 wt% H2O, THI =1.3 and 6.1 wt% H2O, THI = 0.6) suggests water plays a fundamental role in generating the CA fractionation trend. MORB data plot off the trend at a higher THI, possibly related to lower oxygen fugacity during melting and/or crystallization. Models using the pMelts program are consistent with experimentally- and observationally-demonstrated effects of water on suppression of plagioclase and early formation of oxides relative to silicates during magma fractionation, and the resulting FeO* depletion with respect to decreasing MgO.
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Fierens, T; Van Holderbeke, M; Cornelis, C; Jacobs, G; Sioen, I; De Maeyer, M; Vinkx, C; Vanermen, G
2018-07-30
In Europe, 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl)imidazole (THI) and 4-methylimidazole (4-MEI) are - to a certain level - allowed to be present in the food colours ammonia caramel (E 150c) and sulphite ammonia caramel (E 150d). Besides their presence in food colours, exposure to these contaminants may also include other dietary sources. This study describes the occurrence of THI and 4-MEI in a wide variety of food products (n = 522) purchased from the Belgian market and their dietary intake in Belgian consumers from 15 years old onwards. THI was found to be present in 22.4% of the investigated foods at a level up to 551 µg/kg. For 4-MEI (57.7% quantifiable), concentrations up to 2,835 µg/kg were observed. The average dietary intake amounted to 0.02-0.36 µg kg -1 bw -1 day for THI and 0.4-3.7 µg kg -1 bw -1 day for 4-MEI. Coffee, cola and beer were contributing most to the dietary THI and 4-MEI intake in Belgium. Copyright © 2018 Elsevier Ltd. All rights reserved.
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Taveira, Gabriel B; Carvalho, André O; Rodrigues, Rosana; Trindade, Fernanda G; Da Cunha, Maura; Gomes, Valdirene M
2016-01-27
Thionins are a family of plant antimicrobial peptides (AMPs), which participate in plant defense system against pathogens. Here we describe some aspects of the CaThi thionin-like action mechanism, previously isolated from Capsicum annuum fruits. Thionin-like peptide was submitted to antimicrobial activity assays against Candida species for IC50 determination and synergism with fluconazole evaluation. Viability and plasma membrane permeabilization assays, induction of intracellular ROS production analysis and CaThi localization in yeast cells were also investigated. CaThi had strong antimicrobial activity against six tested pathogenic Candida species, with IC50 ranging from 10 to 40 μg.mL(-1). CaThi antimicrobial activity on Candida species was candidacidal. Moreover, CaThi caused plasma membrane permeabilization in all yeasts tested and induces oxidative stresses only in Candida tropicalis. CaThi was intracellularly localized in C. albicans and C. tropicalis, however localized in nuclei in C. tropicalis, suggesting a possible nuclear target. CaThi performed synergistically with fluconazole inhibiting all tested yeasts, reaching 100% inhibition in C. parapsilosis. The inhibiting concentrations for the synergic pair ranged from 1.3 to 4.0 times below CaThi IC50 and from zero to 2.0 times below fluconazole IC50. The results reported herein may ultimately contribute to future efforts aiming to employ this plant-derived AMP as a new therapeutic substance against yeasts.
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Altered proliferation and networks in neural cells derived from idiopathic autistic individuals.
Marchetto, Maria C; Belinson, Haim; Tian, Yuan; Freitas, Beatriz C; Fu, Chen; Vadodaria, Krishna; Beltrao-Braga, Patricia; Trujillo, Cleber A; Mendes, Ana P D; Padmanabhan, Krishnan; Nunez, Yanelli; Ou, Jing; Ghosh, Himanish; Wright, Rebecca; Brennand, Kristen; Pierce, Karen; Eichenfield, Lawrence; Pramparo, Tiziano; Eyler, Lisa; Barnes, Cynthia C; Courchesne, Eric; Geschwind, Daniel H; Gage, Fred H; Wynshaw-Boris, Anthony; Muotri, Alysson R
2017-06-01
Autism spectrum disorders (ASD) are common, complex and heterogeneous neurodevelopmental disorders. Cellular and molecular mechanisms responsible for ASD pathogenesis have been proposed based on genetic studies, brain pathology and imaging, but a major impediment to testing ASD hypotheses is the lack of human cell models. Here, we reprogrammed fibroblasts to generate induced pluripotent stem cells, neural progenitor cells (NPCs) and neurons from ASD individuals with early brain overgrowth and non-ASD controls with normal brain size. ASD-derived NPCs display increased cell proliferation because of dysregulation of a β-catenin/BRN2 transcriptional cascade. ASD-derived neurons display abnormal neurogenesis and reduced synaptogenesis leading to functional defects in neuronal networks. Interestingly, defects in neuronal networks could be rescued by insulin growth factor 1 (IGF-1), a drug that is currently in clinical trials for ASD. This work demonstrates that selection of ASD subjects based on endophenotypes unraveled biologically relevant pathway disruption and revealed a potential cellular mechanism for the therapeutic effect of IGF-1.
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Liu, Daqing; Yovchev, Mladen I.; Zhang, Jinghang; Alfieri, Alan A.; Tchaikovskaya, Tatyana; Laconi, Ezio; Dabeva, Mariana D.
2016-01-01
In normal rat liver, thymocyte antigen 1 (Thy1) is expressed in fibroblasts/myofibroblasts and in some blood progenitor cells. Thy1-expressing cells also accumulate in the liver during impaired liver regeneration. The origin and nature of these cells are not well understood. By using RT-PCR analysis and immunofluorescence microscopy, we describe the presence of rare Thy1+ cells in the liver lobule of normal animals, occasionally forming small collections of up to 20 cells. These cells constitute a small portion (1.7% to 1.8%) of nonparenchymal cells and reveal a mixed mesenchymal-epithelial phenotype, expressing E-cadherin, cytokeratin 18, and desmin. The most potent mitogens for mesenchymal-epithelial Thy1+ cells in vitro are the inflammatory cytokines interferon γ, IL-1, and platelet-derived growth factor-BB, which are not produced by Thy1+ cells. Thy1+ cells express all typical mesenchymal stem cell and hepatic progenitor cell markers and produce growth factor and cytokine mRNA (Hgf, Il6, Tgfa, and Tweak) for proteins that maintain oval cell growth and differentiation. Under appropriate conditions, mesenchymal-epithelial cells differentiate in vitro into hepatocyte-like cells. In this study, we show that the adult rat liver harbors a small pool of endogenous mesenchymal-epithelial cells not recognized previously. In the quiescent state, these cells express both mesenchymal and epithelial cell markers. They behave like hepatic stem cells/progenitors with dual phenotype, exhibiting high plasticity and long-lasting proliferative activity. PMID:25447047
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Houghton, Joanna; Davis, Elaine O.
2012-01-01
Thymidylate synthase (TS) enzymes catalyse the biosynthesis of deoxythymidine monophosphate (dTMP or thymidylate), and so are important for DNA replication and repair. Two different types of TS proteins have been described (ThyA and ThyX), which have different enzymic mechanisms and unrelated structures. Mycobacteria are unusual as they encode both thyA and thyX, and the biological significance of this is not yet understood. Mycobacterium tuberculosis ThyX is thought to be essential and a potential drug target. We therefore analysed M. tuberculosis thyA and thyX expression levels, their essentiality and roles in pathogenesis. We show that both thyA and thyX are expressed in vitro, and that this expression significantly increased within murine macrophages. Under all conditions tested, thyA expression exceeded that of thyX. Mutational studies show that M. tuberculosis thyX is essential, confirming that the enzyme is a plausible drug target. The requirement for M. tuberculosis thyX in the presence of thyA implies that the essential function of ThyX is something other than dTMP synthase. We successfully deleted thyA from the M. tuberculosis genome, and this deletion conferred an in vitro growth defect that was not observed in vivo. Presumably ThyX performs TS activity within M. tuberculosis ΔthyA at a sufficient rate in vivo for normal growth, but the rate in vitro is less than optimal. We also demonstrate that thyA deletion confers M. tuberculosis p-aminosalicylic acid resistance, and show by complementation studies that ThyA T202A and V261G appear to be functional and non-functional, respectively. PMID:22034487
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Fivian-Hughes, Amanda S; Houghton, Joanna; Davis, Elaine O
2012-02-01
Thymidylate synthase (TS) enzymes catalyse the biosynthesis of deoxythymidine monophosphate (dTMP or thymidylate), and so are important for DNA replication and repair. Two different types of TS proteins have been described (ThyA and ThyX), which have different enzymic mechanisms and unrelated structures. Mycobacteria are unusual as they encode both thyA and thyX, and the biological significance of this is not yet understood. Mycobacterium tuberculosis ThyX is thought to be essential and a potential drug target. We therefore analysed M. tuberculosis thyA and thyX expression levels, their essentiality and roles in pathogenesis. We show that both thyA and thyX are expressed in vitro, and that this expression significantly increased within murine macrophages. Under all conditions tested, thyA expression exceeded that of thyX. Mutational studies show that M. tuberculosis thyX is essential, confirming that the enzyme is a plausible drug target. The requirement for M. tuberculosis thyX in the presence of thyA implies that the essential function of ThyX is something other than dTM synthesis [corrected].We successfully deleted thyA from the M. tuberculosis genome, and this deletion conferred an in vitro growth defect that was not observed in vivo. Presumably ThyX performs TS activity within M. tuberculosis ΔthyA at a sufficient rate in vivo for normal growth, but the rate in vitro is less than optimal. We also demonstrate that thyA deletion confers M. tuberculosis p-aminosalicylic acid resistance, and show by complementation studies that ThyA T202A and V261G appear to be functional and non-functional, respectively.
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Vitamin B1 diversity and characterization of biosynthesis genes in cassava.
Mangel, Nathalie; Fudge, Jared B; Fitzpatrick, Teresa B; Gruissem, Wilhelm; Vanderschuren, Hervé
2017-06-15
Vitamin B1, which consists of the vitamers thiamin and its phosphorylated derivatives, is an essential micronutrient for all living organisms because it is required as a metabolic cofactor in several enzymatic reactions. Genetic diversity of vitamin B1 biosynthesis and accumulation has not been investigated in major crop species other than rice and potato. We analyzed cassava germplasm for accumulation of B1 vitamers. Vitamin B1 content in leaves and roots of 41 cassava accessions showed significant variation between accessions. HPLC analyses of B1 vitamers revealed distinct profiles in cassava leaves and storage roots, with nearly equal relative levels of thiamin pyrophosphate and thiamin monophosphate in leaves, but mostly thiamin pyrophosphate in storage roots. Unusually, the cassava genome has two genes encoding the 4-amino-2-methyl-5-hydroxymethylpyrimidine phosphate synthase, THIC (MeTHIC1 and MeTHIC2), both of which carry a riboswitch in the 3'-UTR, as well as the adenylated thiazole synthase, THI1 (MeTHI1a and MeTHI1b). The THIC and THI1 genes are expressed at very low levels in storage roots compared with the accumulation of vitamin B1, indicating only limited biosynthesis de novo therein. In leaves, vitamin B1 content is negatively correlated with THIC and THI1 expression levels, suggesting post-transcriptional regulation of THIC by the riboswitch present in the 3'-UTR of the THIC mRNA and regulation of THI1 by promoter activity or alternative post-transcriptional mechanisms. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.
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Smeets, Bart; Te Loeke, Nathalie A J M; Dijkman, Henry B P M; Steenbergen, Mark L M; Lensen, Joost F M; Begieneman, Mark P V; van Kuppevelt, Toin H; Wetzels, Jack F M; Steenbergen, Eric J
2004-04-01
Focal segmental glomerulosclerosis (FSGS) is a hallmark of progressive renal disease. Podocyte injury and loss have been proposed as the critical events that lead to FSGS. In the present study, the authors have examined the development of FSGS in Thy-1.1 transgenic (tg) mice, with emphasis on the podocyte and parietal epithelial cell (PEC). Thy-1.1 tg mice express the Thy-1.1 antigen on podocytes. Injection of anti-Thy-1.1 mAb induces an acute albuminuria and development of FSGS lesions that resemble human collapsing FSGS. The authors studied FSGS lesions at days 1, 3, 6, 7, 10, 14, and 21, in relation to changes in the expression of specific markers for normal podocytes (WT-1, synaptopodin, ASD33, and the Thy-1.1 antigen), for mouse PEC (CD10), for activated podocytes (desmin), for macrophages (CD68), and for proliferation (Ki-67). The composition of the extracellular matrix (ECM) that forms tuft adhesions or scars was studied using mAb against collagen IV alpha2 and alpha4 chains and antibodies directed against different heparan sulfate species. The first change observed was severe PEC injury at day 1, which increased in time, and resulted in denuded segments of Bowman's capsule at days 6 and 7. Podocytes showed foot process effacement and microvillous transformation. There was no evidence of podocyte loss or denudation of the GBM. Podocytes became hypertrophic at day 3, with decreased expression of ASD33 and synaptopodin and normal expression of WT-1 and Thy-1.1. Podocyte bridges were formed by attachment of hypertrophic podocytes to PEC and podocyte apposition against denuded segments of Bowman's capsule. At day 6, there was a marked proliferation of epithelial cells in Bowman's space. These proliferating cells were negative for desmin and all podocyte markers, but stained for CD10, and thus appeared to be PEC. The staining properties of the early adhesions were identical to that of Bowman's capsule, suggesting that the ECM in the adhesions was produced by PEC. In conclusion, the authors propose the following sequence of events leading to FSGS lesions in the Thy1.1 tg mice: (1) PEC damage and denudation of Bowman's capsule segments; (2) podocyte hypertrophy and bridging; and (3) PEC proliferation with ECM production.
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The response of L5 pyramidal neurons of the PFC to magnetic stimulation from a micro-coil.
Lee, Seung Woo; Fried, Shelley I
2014-01-01
Magnetic stimulation of the nervous system, e.g. transcranial magnetic stimulation (TMS), has been used both to unravel basic structure and function of the nervous system as well as to treat neurological diseases, i.e. clinical depression. Despite progress in both areas, ongoing advancements have been limited by a lack of understanding of the mechanism by which magnetic stimulation alters neural activity. Here, we report responses of cortical neurons to magnetic stimulation arising from a sub-millimeter coil. Cell attached patch clamp was used to record neural activity of layer 5/6 pyramidal neurons of the prefrontal cortex (PFC) in the in vitro mouse brain slice preparation. The fields arising from the small coil were quite different from those arising during clinical TMS but nevertheless allowed the responses of cortical neurons to magnetic stimulation to be probed. For example, the focal nature of induced fields allowed the sensitivity of different regions within targeted pyramidal neurons, e.g. apical dendrite, soma and axon hillock, to be compared. We found that PFC pyramidal neurons were not sensitive to single pulses of stimulation regardless of coil location. However, regions of the apical dendrite and proximal axon were both sensitive to repetitive stimulation as long as the orientation of the induced electric field was aligned with the long axis of the neuron. These results suggest that neurons of the PFC are sensitive to weak magnetic fields and further, that this type of approach may be useful for unraveling some of the mechanisms underlying TMS.
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Martínez-Cáceres, E; Jaleco, A C; Res, P; Noteboom, E; Weijer, K; Spits, H
1998-07-01
In this paper we report that suspensions of human fetal thymocytes contain cells that express high levels of CD34 and Thy-1. These cells were characterized with regard to location within the thymus, phenotype, and function. Confocal laser scan analysis of frozen sections of fetal thymus with anti-CD34 and Thy-1 antibodies revealed that the double-labeled cells were located in the pericortical area. In addition, it was found that the CD34+Thy-1+ cells lacked CD45 and CD50, indicating that these cells are not of hematopoietic origin; this was confirmed by the finding that these cells could be cultured as adherent cells in a medium with cholera toxin and dexamethasone, but failed to grow in mixtures of hematopoietic growth factors. Further analysis indicated that most cultured CD34+Thy-1+ cells expressed cytokeratin (CK) 14 but lacked CK 13, suggesting that these cells are immature epithelial cells. Cultured CD34+Thy-1+ cells were able to induce differentiation of CD1-CD34+CD3-CD4-CD8- thymic precursors into CD4+CD8+ cells in a reaggregate culture in the absence of exogenous cytokines. The CD4+CD8+ cells that developed in these cultures did not express CD3, indicating that CD34+Thy-1+ thymic stromal cells are not capable of completing full T cell differentiation of thymic hematopoietic progenitor cells.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Vallera, D.A.; Youle, R.J.; Neville, D.M. Jr.
1982-03-01
A new method has been devised to eliminate T cells from murine bone marrow grafts across major histocompatibility barriers and thus prevent graft-vs.-host disease (GVHD). The method utilizes a monoclonal antibody directed at the Thy-1.2 antigen but is complement independent. To make anti-Thy-1.2 toxic, the antibody is covalently linked to the toxin ricin. Ricin ordinarily binds, enters, and kills cells through receptors containing galactose. The hybrid protein, anti-Thy-1.2-ricin, can enter and kill cells via the Thy-1.2 receptor. In the presence of lactose the usual entry route for ricin is largely blocked and the hybrid is shown to be a highlymore » selective reagent that is T cell specific in its inhibition of mitogen-stimulated splenocytes. We have used a model of severe and fatal GVHD where BALB/c splenocytes and bone marrow cells are given to irradiated C57BL/6 recipients. Over 90% of these mice die by day 70, exhibiting signs of GVHD. When donor cells are pretreated with 0.5 microgram/ml of anti-Thy-1.2-ricin plus 200 mM lactose before injection, 10 of 11 animals survive through day 70 without signs of GVHD. These studies demonstrate that ricin linked to monoclonal antibodies may have utility related to the prevention of GVHD in human bone marrow transplantation.« less
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Parada, M A; Puig de Parada, M; Hernandez, L; Hoebel, B G
1995-08-01
A low-torque, bubble-free and multiple-channel fluid swivel of easy construction was recently described. This paper describes the design, construction and testing of 3 electrical channels added to the original fluid swivel. The new channels were tested monitoring intrahypothalamic temperature (T(hy)) by means of a copper-constantan thermocouple in freely moving rats, before and after a single intraperitoneal (i.p.) amphetamine injection (3 mg/kg). This test showed an increase in T(hy) after the injection and the maintenance of the electrical continuity along the whole testing period, even when the animals were hyperactive. With this improvement the original swivel was transformed in a more versatile device for experiments requiring fluid handlings and electrophysiological manipulations. Electrical stimulation as in kindling or brain self-stimulation, and electrophysiological recordings as in electroencephalography, electromiography, electrocardiography, in vivo voltammetry and even neuronal unit recording, are just examples of the electrophysiological methods that can be combined with drug self-administration and microdialysis using the present device.
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Parallel pathways from motor and somatosensory cortex for controlling whisker movements in mice
Sreenivasan, Varun; Karmakar, Kajari; Rijli, Filippo M; Petersen, Carl C H
2015-01-01
Mice can gather tactile sensory information by actively moving their whiskers to palpate objects in their immediate surroundings. Whisker sensory perception therefore requires integration of sensory and motor information, which occurs prominently in the neocortex. The signalling pathways from the neocortex for controlling whisker movements are currently poorly understood in mice. Here, we delineate two pathways, one originating from primary whisker somatosensory cortex (wS1) and the other from whisker motor cortex (wM1), that control qualitatively distinct movements of contralateral whiskers. Optogenetic stimulation of wS1 drove retraction of contralateral whiskers while stimulation of wM1 drove rhythmic whisker protraction. To map brainstem pathways connecting these cortical areas to whisker motor neurons, we used a combination of anterograde tracing using adenoassociated virus injected into neocortex and retrograde tracing using monosynaptic rabies virus injected into whisker muscles. Our data are consistent with wS1 driving whisker retraction by exciting glutamatergic premotor neurons in the rostral spinal trigeminal interpolaris nucleus, which in turn activate the motor neurons innervating the extrinsic retractor muscle nasolabialis. The rhythmic whisker protraction evoked by wM1 stimulation might be driven by excitation of excitatory and inhibitory premotor neurons in the brainstem reticular formation innervating both intrinsic and extrinsic muscles. Our data therefore begin to unravel the neuronal circuits linking the neocortex to whisker motor neurons. PMID:25476605
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Predictive modeling targets thymidylate synthase ThyX in Mycobacterium tuberculosis.
Djaout, Kamel; Singh, Vinayak; Boum, Yap; Katawera, Victoria; Becker, Hubert F; Bush, Natassja G; Hearnshaw, Stephen J; Pritchard, Jennifer E; Bourbon, Pauline; Madrid, Peter B; Maxwell, Anthony; Mizrahi, Valerie; Myllykallio, Hannu; Ekins, Sean
2016-06-10
There is an urgent need to identify new treatments for tuberculosis (TB), a major infectious disease caused by Mycobacterium tuberculosis (Mtb), which results in 1.5 million deaths each year. We have targeted two essential enzymes in this organism that are promising for antibacterial therapy and reported to be inhibited by naphthoquinones. ThyX is an essential thymidylate synthase that is mechanistically and structurally unrelated to the human enzyme. DNA gyrase is a DNA topoisomerase present in bacteria and plants but not animals. The current study set out to understand the structure-activity relationships of these targets in Mtb using a combination of cheminformatics and in vitro screening. Here, we report the identification of new Mtb ThyX inhibitors, 2-chloro-3-(4-methanesulfonylpiperazin-1-yl)-1,4-dihydronaphthalene-1,4-dione) and idebenone, which show modest whole-cell activity and appear to act, at least in part, by targeting ThyX in Mtb.
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Endochondral ossification is required for haematopoietic stem-cell niche formation.
Chan, Charles K F; Chen, Ching-Cheng; Luppen, Cynthia A; Kim, Jae-Beom; DeBoer, Anthony T; Wei, Kevin; Helms, Jill A; Kuo, Calvin J; Kraft, Daniel L; Weissman, Irving L
2009-01-22
Little is known about the formation of niches, local micro-environments required for stem-cell maintenance. Here we develop an in vivo assay for adult haematopoietic stem-cell (HSC) niche formation. With this assay, we identified a population of progenitor cells with surface markers CD45(-)Tie2(-)alpha(V)(+)CD105(+)Thy1.1(-) (CD105(+)Thy1(-)) that, when sorted from 15.5 days post-coitum fetal bones and transplanted under the adult mouse kidney capsule, could recruit host-derived blood vessels, produce donor-derived ectopic bones through a cartilage intermediate and generate a marrow cavity populated by host-derived long-term reconstituting HSC (LT-HSC). In contrast, CD45(-)Tie2(-)alpha(V)(+)CD105(+)Thy1(+) (CD105(+)Thy1(+)) fetal bone progenitors form bone that does not contain a marrow cavity. Suppressing expression of factors involved in endochondral ossification, such as osterix and vascular endothelial growth factor (VEGF), inhibited niche generation. CD105(+)Thy1(-) progenitor populations derived from regions of the fetal mandible or calvaria that do not undergo endochondral ossification formed only bone without marrow in our assay. Collectively, our data implicate endochondral ossification, bone formation that proceeds through a cartilage intermediate, as a requirement for adult HSC niche formation.
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Effect of mesenchymal stem cells on anti-Thy1,1 induced kidney injury in albino rats
Sakr, Saber; Rashed, Laila; Zarouk, Waheba; El-Shamy, Rania
2013-01-01
Objective To evaluate the effect of mesenchymal stem cells (MSCs) in rats with anti-Thy1,1 nephritis. Methods Female albino rats were divided into three groups, control group, anti-Thy1,1 group and treatment with i.v. MSCs group. MSCs were derived from bone marrow of male albino rats, Y-chromosome gene was detected by polymerase chain reaction in the kidney. Serum urea and creatinine were estimated for all groups. Kidney of all studied groups was examined histologically and histochemically (total carbohydrates and total proteins). DNA fragmentation and expression of α-SMA were detected. Results Kidney of animals injected with anti-Thy1,1 showed inflammatory leucocytic infiltration, hypertrophied glomeruli, tubular necrosis and congestion in the renal blood vessels. The kidney tissue also showed reduction of carbohydrates and total proteins together with increase in apoptosis and in expression of α-SMA. Moreover, the levels of urea and creatinine were elevated. Treating animals with MSCs revealed that kidney tissue displayed an improvement in the histological and histochemical changes. Apoptosis and α-SMA expression were decreased, and the levels of urea and creatinine decreased. Conclusions The obtained results demonstrated the potential of MSCs to ameliorate the structure and function of the kidney in rats with anti-Thy1,1 nephritis possibly through the release of paracrine growth factor(s). PMID:23620833
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Almandoz-Gil, Leire; Persson, Emma; Lindström, Veronica; Ingelsson, Martin; Erlandsson, Anna; Bergström, Joakim
2018-01-01
The aggregation of alpha-synuclein (αSyn) is the pathological hallmark of Parkinson's disease, dementia with Lewy bodies and related neurological disorders. However, the physiological function of the protein and how this function relates to its pathological effects remain poorly understood. One of the proposed roles of αSyn is to promote the soluble N -ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex assembly by binding to VAMP-2. The objective of this study was to visualize the co-localization between αSyn and the SNARE proteins (VAMP-2, SNAP-25, and syntaxin-1) for the first time using in situ proximity ligation assay (PLA). Cortical primary neurons were cultured from either non-transgenic or transgenic mice expressing human αSyn with the A30P mutation under the Thy-1 promoter. With an antibody recognizing both mouse and human αSyn, a PLA signal indicating close proximity between αSyn and the three SNARE proteins was observed both in the soma and throughout the processes. No differences in the extent of PLA signals were seen between non-transgenic and transgenic neurons. With an antibody specific against human αSyn, the PLA signal was mostly located to the soma and was only present in a few cells. Taken together, in situ PLA is a method that can be used to investigate the co-localization of αSyn and the SNARE proteins in primary neuronal cultures.
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Coexistence of glutamatergic spine synapses and shaft synapses in substantia nigra dopamine neurons
Jang, Miae; Bum Um, Ki; Jang, Jinyoung; Jin Kim, Hyun; Cho, Hana; Chung, Sungkwon; Kyu Park, Myoung
2015-01-01
Dopamine neurons of the substantia nigra have long been believed to have multiple aspiny dendrites which receive many glutamatergic synaptic inputs from several regions of the brain. But, here, using high-resolution two-photon confocal microscopy in the mouse brain slices, we found a substantial number of common dendritic spines in the nigral dopamine neurons including thin, mushroom, and stubby types of spines. However, the number of dendritic spines of the dopamine neurons was approximately five times lower than that of CA1 pyramidal neurons. Immunostaining and morphological analysis revealed that glutamatergic shaft synapses were present two times more than spine synapses. Using local two-photon glutamate uncaging techniques, we confirmed that shaft synapses and spine synapses had both AMPA and NMDA receptors, but the AMPA/NMDA current ratios differed. The evoked postsynaptic potentials of spine synapses showed lower amplitudes but longer half-widths than those of shaft synapses. Therefore, we provide the first evidence that the midbrain dopamine neurons have two morphologically and functionally distinct types of glutamatergic synapses, spine synapses and shaft synapses, on the same dendrite. This peculiar organization could be a new basis for unraveling many physiological and pathological functions of the midbrain dopamine neurons. PMID:26435058
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Kovalev, Grigoriy; Duus, Karen; Wang, Liping; Lee, Robert; Bonyhadi, Mark; Ho, David; McCune, Joseph M.; Kaneshima, Hideto; Su, Lishan
2015-01-01
The SCID-hu Thy/Liv mouse and human fetal thymic organ culture (HF-TOC) models have been used to explore the pathophysiologic mechanisms of HIV-1 infection in the thymus. We report here that HIV-1 infection of the SCID-hu Thy/Liv mouse leads to the induction of MHC class I (MHCI) expression on CD4+CD8+ (DP) thymocytes, which normally express low levels of MHCI. Induction of MHCI on DP thymocytes in HIV-1-infected Thy/Liv organs precedes their depletion and correlates with the pathogenic activity of the HIV-1 isolates. Both MHCI protein and mRNA are induced in thymocytes from HIV-1-infected Thy/Liv organs, indicating induction of MHCI gene expression. Indirect mechanisms are involved, because only a fraction (<10%) of the DP thymocytes were directly infected by HIV-1, although the majority of DP thymocytes are induced to express high levels of MHCI. We further demonstrate that IL-10 is induced in HIV-1-infected thymus organs. Similar HIV-1-mediated induction of MHCI expression was observed in HF-TOC assays. Exogenous IL-10 in HF-TOC induces MHCI expression on DP thymocytes. Therefore, HIV-1 infection of the thymus organ leads to induction of MHCI expression on immature thymocytes via indirect mechanisms involving IL-10. Overexpression of MHCI on DP thymocytes can interfere with thymocyte maturation and may contribute to HIV-1-induced thymocyte depletion. PMID:10358212
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Taveira, Gabriel B; Mello, Érica O; Carvalho, André O; Regente, Mariana; Pinedo, Marcela; de La Canal, Laura; Rodrigues, Rosana; Gomes, Valdirene M
2017-05-01
Many Fusarium species are able to cause severe infections in plants as well as in animals and humans. Therefore, the discovery of new antifungal agents is of paramount importance. CaThi belongs to the thionins, which are cationic peptides with low molecular weights (∼5 kDa) that have toxic effects against various microorganisms. Herein, we study the mechanism of action of CaThi and its combinatory effect with fluconazole (FLC) against Fusarium solani. The mechanism of action of CaThi was studied by growth inhibition, viability, plasma membrane permeabilization, ROS induction, caspase activation, localization, and DNA binding capability, as assessed with Sytox green, DAB, FITC-VAD-FMK, CaThi-FITC, and gel shift assays. The combinatory effect of CaThi and FLC was assessed using a growth inhibition assay. Our results demonstrated that CaThi present a dose dependent activity and at the higher used concentration (50 µg mL -1 ) inhibits 83% of F. solani growth, prevents the formation of hyphae, permeabilizes membranes, induces endogenous H 2 O 2 , activates caspases, and localizes intracellularly. CaThi combined with FLC, at concentrations that alone do not inhibit F. solani, result in 100% death of F. solani when combined. The data presented in this study demonstrate that CaThi causes death of F. solani via apoptosis; an intracellular target may also be involved. Combined treatment using CaThi and FLC is a strong candidate for studies aimed at improved targeting of F. solani. This strategy is of particular interest because it minimizes selection of resistant microorganisms. © 2017 Wiley Periodicals, Inc.
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2004-12-01
were primarily responsible for the organic chemistry and the analytical chemistry, and to Dr. F . Bikker, Mrs. Roos Mars and Mrs. Helma van Dijk for...study as hosts for bacteriophages: TG1 [K-12 A(lac-pro) supE thi hsdD5/ F ’ traD36proA+ B+ laclq lacZ AM 15] and HB2151 [K-12 ara A(lac-pro) thi/ F ’ proA+ B...TG1 [K-12 A(lac-pro) supE thi hsdD5/ F ’ traD36 proA+ B+ lacPq lacZ AM15] and HB2151 [K-12 ara A(lac-pro) thi/ F ’ proA+ B+ laclq Z AM15]. Both strains were
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Mollica, Richard F; Lyoo, In Kyoon; Chernoff, Miriam C; Bui, Hoan X; Lavelle, James; Yoon, Sujung J; Kim, Jieun E; Renshaw, Perry F
2009-11-01
A pilot study of South Vietnamese ex-political detainees who had been incarcerated in Vietnamese reeducation camps and resettled in the United States disclosed significant mental health problems associated with torture and traumatic head injury (THI). To identify structural brain alterations associated with THI and to investigate whether these deficits are associated with posttraumatic stress disorder and depression. Cross-sectional neuroimaging study. Massachusetts General Hospital and McLean Hospital. A subsample of Vietnamese ex-political detainees (n = 42) and comparison subjects (n = 16) selected from a community study of 337 ex-political detainees and 82 comparison subjects. Scores on the Vietnamese versions of the Hopkins Symptom Checklist-25 (HSCL) and Harvard Trauma Questionnaire for depression and posttraumatic stress disorder, respectively; cerebral regional cortical thickness; and manual volumetric morphometry of the amygdala, hippocampus, and thalamus. Ex-political detainees exposed to THI (n = 16) showed a higher rate of depression (odds ratio, 10.2; 95% confidence interval, 1.2-90.0) than those without THI exposure (n = 26). Ex-political detainees with THI had thinner prefrontotemporal cortices than those without THI exposure (P < .001 by the statistical difference brain map) in the left dorsolateral prefrontal and bilateral superior temporal cortices, controlling for age, handedness, and number of trauma/torture events (left superior frontal cortex [SFC], P = .006; left middle frontal cortex, P = .01; left superior temporal cortex [STC], P = .007; right STC, P = .01). Trauma/torture events were associated with bilateral amygdala volume loss (left, P = .045; right, P = .003). Cortical thinning associated with THI in the left SFC and bilateral STC was related to HSCL depression scores in THI-exposed (vs non-THI-exposed) ex-political detainees (left SFC, P for interaction = .007; left STC, P for interaction = .03; right STC, P for interaction = .02). Structural deficits in prefrontotemporal brain regions are linked to THI exposures. These brain lesions are associated with the symptom severity of depression in Vietnamese ex-political detainees.
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Brügemann, K; Gernand, E; von Borstel, U U; König, S
2011-08-01
Data used in the present study included 1,095,980 first-lactation test-day records for protein yield of 154,880 Holstein cows housed on 196 large-scale dairy farms in Germany. Data were recorded between 2002 and 2009 and merged with meteorological data from public weather stations. The maximum distance between each farm and its corresponding weather station was 50 km. Hourly temperature-humidity indexes (THI) were calculated using the mean of hourly measurements of dry bulb temperature and relative humidity. On the phenotypic scale, an increase in THI was generally associated with a decrease in daily protein yield. For genetic analyses, a random regression model was applied using time-dependent (d in milk, DIM) and THI-dependent covariates. Additive genetic and permanent environmental effects were fitted with this random regression model and Legendre polynomials of order 3 for DIM and THI. In addition, the fixed curve was modeled with Legendre polynomials of order 3. Heterogeneous residuals were fitted by dividing DIM into 5 classes, and by dividing THI into 4 classes, resulting in 20 different classes. Additive genetic variances for daily protein yield decreased with increasing degrees of heat stress and were lowest at the beginning of lactation and at extreme THI. Due to higher additive genetic variances, slightly higher permanent environment variances, and similar residual variances, heritabilities were highest for low THI in combination with DIM at the end of lactation. Genetic correlations among individual values for THI were generally >0.90. These trends from the complex random regression model were verified by applying relatively simple bivariate animal models for protein yield measured in 2 THI environments; that is, defining a THI value of 60 as a threshold. These high correlations indicate the absence of any substantial genotype × environment interaction for protein yield. However, heritabilities and additive genetic variances from the random regression model tended to be slightly higher in the THI range corresponding to cows' comfort zone. Selecting such superior environments for progeny testing can contribute to an accurate genetic differentiation among selection candidates. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
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Effects of intraocular injection of a low concentration of zinc on the rat retina.
Nakamichi, N; Chidlow, G; Osborne, N N
2003-10-01
The main aim of this study was to investigate whether intraocular injection of low concentrations of zinc (no greater than 10 microM) aid the survival of ganglion cells in the rat retina after excitotoxic (NMDA) and ischemia/reperfusion injuries. We also determined whether low amounts of zinc cause any detectable retinal toxicity. Intraocular injection of NMDA caused substantial reductions in the mRNA levels of the ganglion cell-specific markers Thy-1 and neurofilament light (NF-L). Co-injection of 0.1 or 1 nmol zinc neither diminished nor exacerbated the effect of NMDA on the levels of these mRNAs. Likewise, ischemia/reperfusion caused significant decreases in the levels of Thy-1 and NF-L mRNAs and in the b-wave amplitude of the electroretinogram. These effects were not counteracted by injection of zinc. Intraocular injection of NMDA caused marked toxicological effects in retinal glial cells, including upregulations of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), glial fibrial acidic protein (GFAP), basic fibroblast growth factor (FGF-2) and ciliary neurotrophic factor (CNTF). Interestingly, injection of 1 nmol zinc caused no changes in the levels of COX-2 and iNOS, yet produced similar, although quantitatively less pronounced, changes in FGF-2, GFAP and CNTF. The upregulations of FGF-2 and CNTF suggest that increasing zinc intake may benefit injured retinal neurons. However, this was not found to be the case in the present studies, perhaps due to the acute nature of the injury paradigms utilised.
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Arena, Salvatore; Latina, Adele; Baratta, Roberto; Burgio, Giuseppe; Gullo, Damiano; Benvenga, Salvatore
2015-04-01
In genetically predisposed individuals, exogenous factors (including pollution) influence the development of Hashimoto's thyroiditis/chronic lymphocytic thyroiditis (CLT). CLT may also be a risk factor for associated thyroid cancer. Few data are available on the role of pollution from petrochemical complexes, one of which is located in the Siracusa province (South-Eastern Sicily), in the pathogenesis of CLT. i) To study the frequency of CLT in fine-needle aspiration cytology (FNAC)-interrogated thyroid nodules from patients who were stably resident in their zones, comparing it in patients living in the petrochemical complex area (zone A) with that of patients from a control area (zone B). ii) To study the frequency of CLT in the FNAC categories of malignancy risk, comparing the two zones. We retrospectively evaluated cytologically adequate slides of 1323 nodules in 1013 outpatients who underwent ultrasound-guided FNAC from 2006 to 2012. We stratified by area of residence, gender, and FNAC categories of malignancy risk. CLT was detected with significantly greater frequency in either patients or nodules from zone A compared with zone B (32.0% vs 23.1%, P=0.002 or 28.2% vs 18.8%, P=0.0001), with a female preponderance (F=35.2% vs M=21.1% or 30.4% vs 20.4%, zone A and F=26.5% vs 12.3% or 21.6% vs 9.5%, zone B). Regardless of zone, CLT was approximately twofold more frequent in the suspiciously malignant+malignant classes (TH4+THY5=47.6%, zone A and 32.4%, zone B) compared with the benign+intermediate classes (THY2+THY3=27.3%, zone A and 18.2%, zone B), but with a clear stepwise THY2 through THY5 increase only in zone A (THY2=25.3%, THY5=66.7%; THY2=18.6%, THY5=28.6% in zone B). The petrochemical complex-related pollution is an environmental factor involved in the development of CLT and, likely, in the CLT association with thyroid neoplasms. © 2015 European Society of Endocrinology.
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Degroote, Roxane L; Hauck, Stefanie M; Treutlein, Gudrun; Amann, Barbara; Fröhlich, Kristina J H; Kremmer, Elisabeth; Merl, Juliane; Stangassinger, Manfred; Ueffing, Marius; Deeg, Cornelia A
2013-12-06
Autoimmune uveitis is characterized by crossing of blood-retinal barrier (BRB) by autoaggressive immune cells. Equine recurrent uveitis (ERU) is a valuable spontaneous model for autoimmune uveitis and analyses of differentially expressed proteins in ERU unraveled changed protein clusters in target tissues and immune system. Healthy eyes are devoid of leukocytes. In ERU, however, leukocytes enter the inner eye and subsequently destroy it. Molecular mechanisms enabling cell migration through BRB still remain elusive. Previously, we detected decreased talin 1 expression in blood-derived granulocytes of ERU cases, linking the innate immune system to ERU. Because changes in leukocyte protein expression pattern may play a role in pathological abnormalities leading to migration ability, we aimed at identifying interactors of talin 1 in leukocytes with immunoprecipitation, followed by LC-MS/MS for candidate identification. This enabled us to identify CD90 (Thy1) as novel interactor of talin 1 besides several other interactors. In blood-derived granulocytes from healthy individuals, CD90 was highly abundant and significantly reduced in ERU, especially in effector cells. Connection between talin 1 and CD90 and their expression differences in inflammation is an interesting novel finding allowing deeper insight into immune response of innate immune system and granulocyte migration ability in this organ-specific autoimmune disease.
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Ultramicroscopy as a novel tool to unravel the tropism of AAV gene therapy vectors in the brain.
Alves, Sandro; Bode, Julia; Bemelmans, Alexis-Pierre; von Kalle, Christof; Cartier, Nathalie; Tews, Björn
2016-06-20
Recombinant adeno-associated viral (AAV) vectors have advanced to the vanguard of gene therapy. Numerous naturally occurring serotypes have been used to target cells in various tissues. There is a strong need for fast and dynamic methods which efficiently unravel viral tropism in whole organs. Ultramicroscopy (UM) is a novel fluorescence microscopy technique that images optically cleared undissected specimens, achieving good resolutions at high penetration depths while being non-destructive. UM was applied to obtain high-resolution 3D analysis of AAV transduction in adult mouse brains, especially in the hippocampus, a region of interest for Alzheimer's disease therapy. We separately or simultaneously compared transduction efficacies for commonly used serotypes (AAV9 and AAVrh10) using fluorescent reporter expression. We provide a detailed comparative and quantitative analysis of the transduction profiles. UM allowed a rapid analysis of marker fluorescence expression in neurons with intact projections deep inside the brain, in defined anatomical structures. Major hippocampal neuronal transduction was observed with both vectors, with slightly better efficacy for AAV9 in UM. Glial response and synaptic marker expression did not change post transduction.We propose UM as a novel valuable complementary tool to efficiently and simultaneously unravel tropism of different viruses in a single non-dissected adult rodent brain.
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Accumulation of oligomer-prone α-synuclein exacerbates synaptic and neuronal degeneration in vivo
Rockenstein, Edward; Nuber, Silke; Overk, Cassia R.; Ubhi, Kiren; Mante, Michael; Patrick, Christina; Adame, Anthony; Trejo-Morales, Margarita; Gerez, Juan; Picotti, Paola; Jensen, Poul H.; Campioni, Silvia; Riek, Roland; Winkler, Jürgen; Gage, Fred H.; Winner, Beate
2014-01-01
In Parkinson’s disease and dementia with Lewy bodies, α-synuclein aggregates to form oligomers and fibrils; however, the precise nature of the toxic α-synuclein species remains unclear. A number of synthetic α-synuclein mutations were recently created (E57K and E35K) that produce species of α-synuclein that preferentially form oligomers and increase α-synuclein-mediated toxicity. We have shown that acute lentiviral expression of α-synuclein E57K leads to the degeneration of dopaminergic neurons; however, the effects of chronic expression of oligomer-prone α-synuclein in synapses throughout the brain have not been investigated. Such a study could provide insight into the possible mechanism(s) through which accumulation of α-synuclein oligomers in the synapse leads to neurodegeneration. For this purpose, we compared the patterns of neurodegeneration and synaptic damage between a newly generated mThy-1 α-synuclein E57K transgenic mouse model that is prone to forming oligomers and the mThy-1 α-synuclein wild-type mouse model (Line 61), which accumulates various forms of α-synuclein. Three lines of α-synuclein E57K (Lines 9, 16 and 54) were generated and compared with the wild-type. The α-synuclein E57K Lines 9 and 16 were higher expressings of α-synuclein, similar to α-synuclein wild-type Line 61, and Line 54 was a low expressing of α-synuclein compared to Line 61. By immunoblot analysis, the higher-expressing α-synuclein E57K transgenic mice showed abundant oligomeric, but not fibrillar, α-synuclein whereas lower-expressing mice accumulated monomeric α-synuclein. Monomers, oligomers, and fibrils were present in α-synuclein wild-type Line 61. Immunohistochemical and ultrastructural analyses demonstrated that α-synuclein accumulated in the synapses but not in the neuronal cells bodies, which was different from the α-synuclein wild-type Line 61, which accumulates α-synuclein in the soma. Compared to non-transgenic and lower-expressing mice, the higher-expressing α-synuclein E57K mice displayed synaptic and dendritic loss, reduced levels of synapsin 1 and synaptic vesicles, and behavioural deficits. Similar alterations, but to a lesser extent, were seen in the α-synuclein wild-type mice. Moreover, although the oligomer-prone α-synuclein mice displayed neurodegeneration in the frontal cortex and hippocampus, the α-synuclein wild-type only displayed neuronal loss in the hippocampus. These results support the hypothesis that accumulating oligomeric α-synuclein may mediate early synaptic pathology in Parkinson’s disease and dementia with Lewy bodies by disrupting synaptic vesicles. This oligomer-prone model might be useful for evaluating therapies directed at oligomer reduction. PMID:24662516
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Accumulation of oligomer-prone α-synuclein exacerbates synaptic and neuronal degeneration in vivo.
Rockenstein, Edward; Nuber, Silke; Overk, Cassia R; Ubhi, Kiren; Mante, Michael; Patrick, Christina; Adame, Anthony; Trejo-Morales, Margarita; Gerez, Juan; Picotti, Paola; Jensen, Poul H; Campioni, Silvia; Riek, Roland; Winkler, Jürgen; Gage, Fred H; Winner, Beate; Masliah, Eliezer
2014-05-01
In Parkinson's disease and dementia with Lewy bodies, α-synuclein aggregates to form oligomers and fibrils; however, the precise nature of the toxic α-synuclein species remains unclear. A number of synthetic α-synuclein mutations were recently created (E57K and E35K) that produce species of α-synuclein that preferentially form oligomers and increase α-synuclein-mediated toxicity. We have shown that acute lentiviral expression of α-synuclein E57K leads to the degeneration of dopaminergic neurons; however, the effects of chronic expression of oligomer-prone α-synuclein in synapses throughout the brain have not been investigated. Such a study could provide insight into the possible mechanism(s) through which accumulation of α-synuclein oligomers in the synapse leads to neurodegeneration. For this purpose, we compared the patterns of neurodegeneration and synaptic damage between a newly generated mThy-1 α-synuclein E57K transgenic mouse model that is prone to forming oligomers and the mThy-1 α-synuclein wild-type mouse model (Line 61), which accumulates various forms of α-synuclein. Three lines of α-synuclein E57K (Lines 9, 16 and 54) were generated and compared with the wild-type. The α-synuclein E57K Lines 9 and 16 were higher expressings of α-synuclein, similar to α-synuclein wild-type Line 61, and Line 54 was a low expressing of α-synuclein compared to Line 61. By immunoblot analysis, the higher-expressing α-synuclein E57K transgenic mice showed abundant oligomeric, but not fibrillar, α-synuclein whereas lower-expressing mice accumulated monomeric α-synuclein. Monomers, oligomers, and fibrils were present in α-synuclein wild-type Line 61. Immunohistochemical and ultrastructural analyses demonstrated that α-synuclein accumulated in the synapses but not in the neuronal cells bodies, which was different from the α-synuclein wild-type Line 61, which accumulates α-synuclein in the soma. Compared to non-transgenic and lower-expressing mice, the higher-expressing α-synuclein E57K mice displayed synaptic and dendritic loss, reduced levels of synapsin 1 and synaptic vesicles, and behavioural deficits. Similar alterations, but to a lesser extent, were seen in the α-synuclein wild-type mice. Moreover, although the oligomer-prone α-synuclein mice displayed neurodegeneration in the frontal cortex and hippocampus, the α-synuclein wild-type only displayed neuronal loss in the hippocampus. These results support the hypothesis that accumulating oligomeric α-synuclein may mediate early synaptic pathology in Parkinson's disease and dementia with Lewy bodies by disrupting synaptic vesicles. This oligomer-prone model might be useful for evaluating therapies directed at oligomer reduction.
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Emodin down-regulates expression of TRPV1 mRNA and its function in DRG neurons in vitro.
Sui, Feng; Huo, Hai-Ru; Zhang, Chang-Bin; Yang, Na; Guo, Jian-You; Du, Xin-Liang; Zhao, Bao-Sheng; Liu, Hong-Bin; Li, Lan-Fang; Guo, Shu-Ying; Jiang, Ting-Liang
2010-01-01
Emodin is a principle ingredient isolated from rhubarb rhizome, which is commonly used for constipation or pain-related diseases in traditional Chinese medicine (TCM) practice. The transient receptor potential vanilloid 1 ion channel proteins (TRPV1) are abundantly expressed in the peripheral sensory neurons and are assumed to act as a kind of nociceptor involved in the perception of pain and development of hyperalgesia. The aim of this study was to further unravel the analgesic mechanisms of rhubarb through investigating the effects of its main constitutive ingredient emodin on the expression of TRPV1 mRNA as well as on its calcium- mediating functions in vitro. The primary DRG neurons with a high purity and viability were obtained, and the TRPV1 mRNA expression levels were examined by using real-time RT-PCR and the elevated amplitudes of intracellular [Ca(2+)]i in the DRG neurons evoked by TRPV1 agonist capsaicin were examined by confocal microscopy. The results showed that emodin could significantly down-regulate both the mRNA expression of TRPV1 and the capsaicin-evoked intracellular fluorescent intensity in the DRG neurons under both 37 degrees C and 39 degrees C in vitro. Concomitantly, all of the changes induced by emodin could not be blocked by pretreatment of the primary neurons with capsazepine, an antagonist of TRPV1. In conclusion, we established that the mRNA expression level of TRPV1 and its calcium-mediating function in naive DRG neurons could be down-regulated by emodin through perhaps the non-TRPV1 channel pathways, and this might be the molecular mechanisms for rhubarb to inhibit hyperalgesia induced by inflammatory stimuli.
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NASA Astrophysics Data System (ADS)
Menegassi, Silvio Renato Oliveira; Pereira, Gabriel Ribas; Dias, Eduardo Antunes; Koetz, Celso; Lopes, Flávio Guiselli; Bremm, Carolina; Pimentel, Concepta; Lopes, Rubia Branco; da Rocha, Marcela Kuczynski; Carvalho, Helena Robattini; Barcellos, Júlio Otavio Jardim
2016-01-01
The objective of this study was to evaluate the seasonal effects of the environment on sperm quality in subtropical region determined by temperature and humidity index (THI). We used 20 Brangus bulls (5/8 Angus × 3/8 Nellore) aged approximately 24 months at the beginning of the study. Semen evaluations were performed twice per season during 1 year. Climate THI data were collected from an automatic weather station from the National Institute of Meteorology. Infrared thermography images were used to determine the temperature of the proximal and distal poles of the testis to assess the testicular temperature gradient (TG). The seasonal effects on seminal and climatic variables were analyzed with ANOVA using MIXED procedure of SAS. Sperm motility in spring (60.1 %), summer (57.6 %), and autumn (64.5 %) showed difference compared to winter (73.0 %; P < 0.01). TG was negatively correlated with THI at 18 days (spermiogenesis) (-0.76; P < 0.05) and at 12 days (epididymal transit) (-0.85; P < 0.01). Ocular temperature (OcT) had a positive correlation with THI at 18 days (0.78; P < 0.05) and at 12 days (0.84; P < 0.01). Motility showed a negative correlation with THI only at 18 days (-0.79; P < 0.05). During spermiogenesis, the TG had higher negative correlation compared to OcT (-0.97; P < 0.01) and rectal temperature (-0.72; P < 0.05). Spermatozoa with distal midpiece reflex were correlated with THI during transit epididymis (0.72; P < 0.05). Seminal parameters are not affected when THI reaches 93.0 (spermiogenesis) and 88.0 (epididymal transit). We concluded that infrared thermography can be adopted as an indirect method in order to assess the effect of environmental changes in TG and OcT of Brangus bulls.
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Wilde, Marcelo L; Menz, Jakob; Trautwein, Christoph; Leder, Christoph; Kümmerer, Klaus
2016-06-01
An experimental and in silico quantitative structure-activity relationship (QSAR) approach was applied to assess the environmental fate and effects of the antipsychotic drug Thioridazine (THI). The sunlight-driven attenuation of THI was simulated using a Xenon arc lamp. The photodegradation reached the complete primary elimination, whereas 97% of primary elimination and 11% of mineralization was achieved after 256 min of irradiation for the initial concentrations of 500 μg L(-1) and 50 mg L(-1), respectively. A non-target approach for the identification and monitoring of transformation products (TPs) was adopted. The structure of the TPs was further elucidated using liquid chromatography-high resolution mass spectrometry (LC-HRMS). The proposed photodegradation pathway included sulfoxidation, hydroxylation, dehydroxylation, and S- and N-dealkylation, taking into account direct and indirect photolysis through a self-sensitizing process in the higher concentration studied. The biodegradability of THI and photolytic samples of THI was tested according to OECD 301D and 301F, showing that THI and the mixture of TPs were not readily biodegradable. Furthermore, THI was shown to be highly toxic to environmental bacteria using a modified luminescent bacteria test with Vibrio fischeri. This bacteriotoxic activity of THI was significantly reduced by phototransformation and individual concentration-response analysis confirmed a lowered bacterial toxicity for the sulfoxidation products Thioridazine-2-sulfoxide and Thioridazine-5-sulfoxide. Additionally, the applied QSAR models predicted statistical and rule-based positive alerts of mutagenic activities for carbazole derivative TPs (TP 355 and TP 339) formed through sulfoxide elimination, which would require further confirmatory in vitro validation tests. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Effect of thermal stress on fertility and egg quality of Japanese quail.
El-Tarabany, Mahmoud S
2016-10-01
Heat stress is one of the major causes of a decreased performance of laying quail in tropical and subtropical countries. The aim of this study was to investigate the impact of temperature humidity index (THI) on fertility aspects, external and internal egg quality parameters in Japanese quail. One hundred and forty four (144) Japanese quail, 12 of weeks age, were used. Birds were divided randomly into three equal groups, control (at low THI, lower than 70), H 1 (at moderate THI, 70-75) and H 2 (at high THI, 76-80). Quail in the control and H 1 groups had significant greater fertility (p=0.021) and hatchability % (p=0.037), compared with H 2 group. Quail in the control group (at low THI) laid heavier egg weight with a higher external (egg weight (p=0.03), shell thickness, shell weight, eggshell ratio and eggshell density (p=0.001)) and internal egg quality score (albumin weight (p=0.026), yolk height (p=0.003), yolk index (p=0.039) and Haugh unit (p=0.001)). Otherwise, such quality traits were compromised in heat-stressed quail. At the high THI level, egg weight had a significant positive correlation with albumin weight (r=0.58, p<0.01), yolk weight (r=0.22, p<0.05), albumen ratio (r=0.17, p<0.05), yolk height (r=0.14, p<0.05) and yolk index (r=0.18, p<0.05), but was negatively correlated with yolk ratio (r=-0.15, p<0.05). Japanese quail exposed to heat stress (THI over 75) revealed drop in fertility indices and egg quality traits, indicating a detrimental policy of economic income. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Chen, Danping; Wang, Xiaoqian; Huang, Hongming; Fu, Min; Ge, Runmei; Wu, Peina; Cui, Yong
2015-11-01
To investigate the correlation betwen varying degrees of non-acute tinnitus and anxiety and depression. Seventy-seven outpatients with non-acute tinnitus as their in chief complaint were submitted to Tinnitus Handicap Inventory(THI), Self-Rating Anxiety Scale(SAS), and Self-Rating Depression Scale (SDS). THI and its three subscales were found to have significant correlations with SAS and SDS. The group (THI ≥ 38) had more anxiety and depression than the mild (THI < 38). Significant correlations were also observed between THI, SAS and SDS in the group with THI ≥ 38, in contrast with the group of THI < 38. Patients with THI ≥ 38 suffered from severe anxiety and depression than the mild. Doctors should pay more attention to these patients, especially their psychological disorders.
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Ward, Patricia J; Jones, Laura N; Mulligan, Amanda; Goolsby, William; Wilhelm, Jennifer C; English, Arthur W
2016-01-01
Peripheral nerve injuries are common, and functional recovery is very poor. Beyond surgical repair of the nerve, there are currently no treatment options for these patients. In experimental models of nerve injury, interventions (such as exercise and electrical stimulation) that increase neuronal activity of the injured neurons effectively enhance axon regeneration. Here, we utilized optogenetics to determine whether increased activity alone is sufficient to promote motor axon regeneration. In thy-1-ChR2/YFP transgenic mice in which a subset of motoneurons express the light-sensitive cation channel, channelrhodopsin (ChR2), we activated axons in the sciatic nerve using blue light immediately prior to transection and surgical repair of the sciatic nerve. At four weeks post-injury, direct muscle EMG responses evoked with both optical and electrical stimuli as well as the ratio of these optical/electrical evoked EMG responses were significantly greater in mice that received optical treatment. Thus, significantly more ChR2+ axons successfully re-innervated the gastrocnemius muscle in mice that received optical treatment. Sections of the gastrocnemius muscles were reacted with antibodies to Synaptic Vesicle Protein 2 (SV2) to quantify the number of re-occupied motor endplates. The number of SV2+ endplates was greater in mice that received optical treatment. The number of retrogradely-labeled motoneurons following intramuscular injection of cholera toxin subunit B (conjugated to Alexa Fluor 555) was greater in mice that received optical treatment. Thus, the acute (1 hour), one-time optical treatment resulted in robust, long-lasting effects compared to untreated animals as well as untreated axons (ChR2-). We conclude that neuronal activation is sufficient to promote motor axon regeneration, and this regenerative effect is specific to the activated neurons.
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Reliability and Validity of the Chinese (Mandarin) Tinnitus Handicap Inventory
Meng, Zhaoli; Zheng, Yun; Wang, Kai; Kong, Xiudan; Tao, Yong; Xu, Ke; Liu, Guanjian
2012-01-01
Objectives The Tinnitus Handicap Inventory (THI) is a commonly used self-reporting tinnitus questionnaire. We undertook this study to determine the reliability and validity of the Chinese-Mandarin version of the Tinnitus Handicap Inventory (THI-CM) for measuring tinnitus-related handicaps. Methods We tested the test-retest reliability, internal reliability, and construct validity of the THI-CM. Two-hundred patients seeking treatment for primary or secondary tinnitus in Southwest China were asked to complete THI-CM prior to clinical evaluation. Patients were evaluated by a clinician using standard methods, and 40 patients were asked to complete THI-CM a second time 14±3 days after the initial interview. Results The test-retest reliability of THI-CM was high (Pearson correlation, 0.98), as was the internal reliability (Cronbach's α, 0.93). Factor analysis indicated that THI-CM has a unifactorial structure. Conclusion The THI-CM version is reliable. The total score in THI-CM can be used to measure tinnitus-related handicaps in Mandarin-speaking populations. PMID:22468196
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Is reticular temperature a useful indicator of heat stress in dairy cattle?
Ammer, S; Lambertz, C; Gauly, M
2016-12-01
The present study investigated whether reticular temperature (RT) in dairy cattle is a useful indicator of heat stress considering the effects of milk yield and water intake (WI). In total, 28 Holstein-Friesian dairy cows raised on 3 farms in Lower Saxony, Germany, were studied from March to December 2013. During the study, RT and barn climate parameters (air temperature, relative humidity) were measured continuously and individual milk yield was recorded daily. Both the daily temperature-humidity index (THI) and the daily median RT per cow were calculated. Additionally, the individual WI (amount and frequency) of 10 cows during 100d of the study was recorded on 1 farm. Averaged over all farms, daily THI ranged between 35.4 and 78.9 with a mean (±standard deviation) of 60.2 (±8.7). Dairy cows were on average (±standard deviation) 110.9d in milk (±79.3) with a mean (±standard deviation) milk yield of 35.2kg/d (±9.1). The RT was affected by THI, milk yield, days in milk, and WI. Up to a THI threshold of 65, RT remained constant at 39.2°C. Above this threshold, RT increased to 39.3°C and further to 39.4°C when THI ≥70. The correlation between THI ≥70 and RT was 0.22, whereas the coefficient ranged between r=-0.08 to +0.06 when THI <70. With increasing milk yield, RT decreased slightly from 39.3°C (<30kg/d) to 39.2°C (≥40kg/d). For daily milk yields of ≥40kg, the median RT and daily milk yield were correlated at r=-0.18. The RT was greater when dairy cows yielded ≥30kg/d and THI ≥70 (39.5°C) compared with milk yields <30kg and THI <70 (39.3°C). The WI, which averaged (±standard deviation) 11.5 l (±5.7) per drinking bout, caused a mean decrease in RT of 3.2°C and was affected by the amount of WI (r=0.60). After WI, it took up to 2h until RT reached the initial level before drinking. In conclusion, RT increased when the THI threshold of 65 was exceeded. A further increase was noted when THI ≥70. Nevertheless, the effects of WI and milk yield have to be considered carefully when RT is used to detect hyperthermia in dairy cattle. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
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Kim, So Young; Chang, Mun Young; Hong, Min; Yoo, Sun-Gil; Oh, Dongik; Park, Moo Kyun
2017-10-01
Notched music therapy has been suggested to be effective for relieving tinnitus. We have developed a smartphone application using tailor-made notched music for tinnitus patients. This study aimed to evaluate the effect of this smartphone application on reducing tinnitus. In addition, we investigated the predictive factors for tinnitus treatment outcome using this smartphone application. A total of 26 patients who were chronically distressed by tinnitus with a ≥18 Tinnitus Handicap Inventory (THI) score were recruited from March 2013 to March 2015 (National Clinical Trial (NCT) Identifier Number 01663467). Patients were instructed to listen to tailor-made notched music through our smartphone application for 30-60min per day and were prescribed Ginkgo biloba for 3 months. Treatment outcome was evaluated using the THI, a visual analogue scale that measures the effects of tinnitus in terms of loudness, noticeable time, annoyance, and disruption of daily life. Demographic data, including age, sex, duration of tinnitus, and pre-treatment scores on questionnaires such as the Beck Depression Inventory (BDI), State Trait Anxiety Inventory (TAI), and Pittsburgh Sleep Quality Index (PSQI) scores were compared between the effective and non-effective groups according to the differences between their pre- and post-treatment THI scores. Smartphone application-delivered notched music therapy and Ginko combined treatment improved the THI score from 33.9±18.9 to 23.1±15.2; the effect was particularly marked for the emotional score of the THI. Improvement in the THI score was positively correlated with the initial THI score (P=0.001, adjusted estimated value=0.49, 95% confidence interval=0.25-0.73). Chronic tinnitus patients who underwent smartphone application-delivered notched music therapy and Ginko combined treatment showed improved THI scores, particularly the emotional score of the THI. A smartphone application-delivered therapy and Ginko combined treatment may be more effective in tinnitus patients who have had a higher initial THI. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
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Mobile group II intron based gene targeting in Lactobacillus plantarum WCFS1.
Sasikumar, Ponnusamy; Paul, Eldho; Gomathi, Sivasamy; Abhishek, Albert; Sasikumar, Sundaresan; Selvam, Govindan Sadasivam
2016-10-01
The usage of recombinant lactic acid bacteria for delivery of therapeutic proteins to the mucosa has been emerging. In the present study, an attempt was made to engineer a thyA mutant of Lactobacillus plantarum (L. plantarum) using lactococcal group II intron Ll.LtrB for the development of biologically contained recombinant L. plantarum for prevention of calcium oxalate stone disease. The 3 kb Ll.LtrB intron donor cassettes from the source vector pACD4C was PCR amplified, ligated into pSIP series of lactobacillus vector pLp_3050sAmyA, yielding a novel vector pLpACD4C (8.6 kb). The quantitative real-time PCR experiment shows 94-fold increased expression of Ll.LtrB intron and 14-fold increased expression of ltrA gene in recombinant L. plantarum containing pLpACD4C. In order to target the thyA gene, the potential intron RNA binding sites in the thyA gene of L. plantarum was predicted with help of computer algorithm. The insertion location 188|189s of thyA gene (lowest E-0.134) was chosen and the wild type intron Ll.LtrB was PCR modified, yielding a retargeted intron of pLpACDthyA. The retargeted intron was expressed by using induction peptide (sppIP), subsequently the integration of intron in thyA gene was identified by PCR screening and finally ThyA - mutant of L. plantarum (ThyA18) was detected. In vitro growth curve result showed that in the absence of thymidine, colony forming units of mutant ThyA18 was decreased, whereas high thymidine concentration (10 μM) supported the growth of the culture until saturation. In conclusion, ThyA - mutant of L. plantarum (ThyA18) constructed in this study will be used as a biologically contained recombinant probiotic to deliver oxalate decarboxylase into the lumen for treatment of hyperoxaluria and calcium oxalate stone deposition. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Pettoello-Mantovani, M; Kollmann, T R; Raker, C; Kim, A; Yurasov, S; Tudor, R; Wiltshire, H; Goldstein, H
1997-01-01
Treatment with protease inhibitors alone or in combination with inhibitors of reverse transcriptase potently suppresses levels of human immunodeficiency virus (HIV) RNA in plasma and thereby may significantly delay the progression of HIV-mediated disease. To investigate the effect of treatment with the protease inhibitor saquinavir on HIV replication in the lymphoid tissues, we used a SCID-hu mouse model that we developed, in which human thymic and liver tissues (hu-thy/liv) were implanted under both kidney capsules in SCID mice (thy/liv-SCID-hu mice). These mice are populated in the periphery with large numbers of human T cells and develop disseminated HIV infection after intraimplant injection. thy/liv-SCID-hu mice with established HIV infection that were treated for 1 month with saquinavir had a significantly lower viral load present in the implanted hu-thy/liv and mouse spleen than did the untreated HIV-infected thy/liv-SCID-hu mice. To examine the capacity of acute treatment with saquinavir to prevent HIV infection, some thy/liv-SCID-hu mice were inoculated with HIV and then immediately started on saquinavir. Although treated mice had markedly lower viral loads in the thy/liv implants and spleens, HIV infection was not completely prevented. Thus, the effect of antiviral therapy on HIV infection in the major site of HIV replication, the lymphoid tissues, can be readily evaluated in our thy/liv-SCID-hu mice. These mice should prove to be a useful model for determining the in vivo effectiveness of different therapeutic interventions on acute and chronic HIV infection. PMID:9303378
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Lee, Jae Hee; Ra, Jin-Ju; Kim, Young Ho
2014-04-01
The Tinnitus Handicap Inventory (THI) often requires patients to devote their time to complete the questionnaire than they expect. Given that it limits the effectiveness of THI in a busy clinical practice that desires a quick and easy assessment of tinnitus handicap, this study evaluated clinical usefulness of a Simplified version of Tinnitus Handicap Inventory (THI-S) in measuring the severity of tinnitus handicap as well as predicting the psychological distress associated with tinnitus. A total of 129 outpatients suffering from tinnitus (61 with normal hearing and 68 with hearing loss) participated in this study. The responses of THI-S (10 items) and THI (25 items) were evaluated to quantify the subjective handicap of tinnitus. The self-perceived level of stress, anxiety, and depression of all participants was measured with a series of self-report questionnaires such as Korean version of Brief Encounter Psychosocial Instrument (BEPSI), State Trait Anxiety Inventory (STAI), and Beck Depression Inventory (BDI), respectively. All the questionnaire responses were analyzed using non-parametric analyses to examine the convergence, comparability, internal consistency reliability and validity of THI-S. The severity of tinnitus handicap and the relevant psychological distress greatly varied across individuals. The THI-S responses were comparable to original THI responses, regardless of hearing status of patients. The internal consistency of THI-S responses was found to be good for total score (Cronbach's α=0.83-0.91), with moderately high consistency for the emotional, functional, and catastrophic subscales. Significant (p<0.01) correlations of the THI-S with the THI (rs =0.95) as well as with the BEPSI, STAI, BDI questionnaires (rs =0.40-0.45) suggest that the THI-S questionnaire appeared to be useful to provide objective data of subjective tinnitus handicap as well as predict psychological distress. Three factors were extracted through factor analysis, which explained 73% of the total variance. Despite reducing the length of 25 items of THI to 10 items, the THI-S can be used as s a simple but reliable and valid tool for evaluating severity of tinnitus handicap as well as detecting its negative impact on psychological functioning.
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El-Tarabany, Mahmoud S; El-Tarabany, Akram A; Atta, Mostafa A
2017-01-01
The objective of this study was to evaluate the impact of thermal stress on milk production and physiological traits of Baladi goats under subtropical Egyptian conditions. Sixty dairy Baladi goats were exposed to three different levels of temperature-humidity index (THI), including low (less than 70), moderate (over 70 and up to 80), and high levels (over 80). The influence of THI on the milk composition and physiological, hematological, and biochemical traits was investigated. Rectal temperature and respiration rate were significantly greater at the higher THI than at low and moderate THI (p = 0.016 and 0.002, respectively). Baladi goats had decreased daily milk yield in a rate of 27.3 and 19.3 % at high THI level, compared with low and moderate THI, respectively (p = 0.031). On the contrary, no significant differences have been reported in protein, fat, and total solids percentages at different THI levels. Total leucocyte count, serum glucose, and total protein were significantly reduced at high THI in comparison with low and moderate THI levels (p = 0.043, 0.001, and 0.001, respectively). However, dairy goats maintained relatively stable estimates for erythrocytes count, hemoglobin, serum triglycerides, cholesterol, catalase, total antioxidant capacity, and triiodothyronine at different THI levels. Our results indicate that dairy Baladi goats can tolerate THI levels up to 80; however, variable reduction in milk yield and few biochemical (serum total protein and glucose) and hematological (leucocytes count) parameters have been reported at a THI level higher than 80.
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Myosin IIA-related Actomyosin Contractility Mediates Oxidative Stress-induced Neuronal Apoptosis
Wang, Yan; Xu, Yingqiong; Liu, Qian; Zhang, Yuanyuan; Gao, Zhen; Yin, Mingzhu; Jiang, Nan; Cao, Guosheng; Yu, Boyang; Cao, Zhengyu; Kou, Junping
2017-01-01
Oxidative stress-induced neuronal apoptosis plays an important role in the progression of central nervous system (CNS) diseases. In our study, when neuronal cells were exposed to hydrogen peroxide (H2O2), an exogenous oxidant, cell apoptosis was observed with typical morphological changes including membrane blebbing, neurite retraction and cell contraction. The actomyosin system is considered to be responsible for the morphological changes, but how exactly it regulates oxidative stress-induced neuronal apoptosis and the distinctive functions of different myosin II isoforms remain unclear. We demonstrate that myosin IIA was required for neuronal contraction, while myosin IIB was required for neuronal outgrowth in normal conditions. During H2O2-induced neuronal apoptosis, myosin IIA, rather than IIB, interacted with actin filaments to generate contractile forces that lead to morphological changes. Moreover, myosin IIA knockout using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein-9 nuclease (CRISPR/Cas9) reduced H2O2-induced neuronal apoptosis and the associated morphological changes. We further demonstrate that caspase-3/Rho-associated kinase 1 (ROCK1) dependent phosphorylation of myosin light chain (MLC) was required for the formation of the myosin IIA-actin complex. Meanwhile, either inhibition of myosin II ATPase with blebbistatin or knockdown of myosin IIA with siRNA reversely attenuated caspase-3 activation, suggesting a positive feedback loop during oxidative stress-induced apoptosis. Based on our observation, myosin IIA-actin complex contributes to actomyosin contractility and is associated with the positive feedback loop of caspase-3/ROCK1/MLC pathway. This study unravels the biochemical and mechanistic mechanisms during oxidative stress-induced neuronal apoptosis and may be applicable for the development of therapies for CNS diseases. PMID:28352215
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Neuronal dysfunction with aging and its amelioration
ANDO, Susumu
2012-01-01
The author focused on the functional decline of synapses in the brain with aging to understand the underlying mechanisms and to ameliorate the deficits. The first attempt was to unravel the neuronal functions of gangliosides so that gangliosides could be used for enhancing synaptic activity. The second attempt was to elicit the neuronal plasticity in aged animals through enriched environmental stimulation and nutritional intervention. Environmental stimuli were revealed neurochemically and morphologically to develop synapses leading to enhanced cognitive function. Dietary restriction as a nutritional intervention restored the altered metabolism of neuronal membranes with aging, providing a possible explanation for the longevity effect of dietary restriction. These results obtained with aging and dementia models of animals would benefit aged people. PMID:22728441
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Spencer, Brian; Valera, Elvira; Rockenstein, Edward; Overk, Cassia; Mante, Michael; Adame, Anthony; Zago, Wagner; Seubert, Peter; Barbour, Robin; Schenk, Dale; Games, Dora; Rissman, Robert A; Masliah, Eliezer
2017-01-13
Neurodegenerative disorders such as Parkinson's Disease (PD), PD dementia (PDD) and Dementia with Lewy bodies (DLB) are characterized by progressive accumulation of α-synuclein (α-syn) in neurons. Recent studies have proposed that neuron-to-neuron propagation of α-syn plays a role in the pathogenesis of these disorders. We have previously shown that antibodies against the C-terminus of α-syn reduce the intra-neuronal accumulation of α-syn and related deficits in transgenic models of synucleinopathy, probably by abrogating the axonal transport and accumulation of α-syn in in vivo models. Here, we assessed the effect of passive immunization against α-syn in a new mouse model of axonal transport and accumulation of α-syn. For these purpose, non-transgenic, α-syn knock-out and mThy1-α-syn tg (line 61) mice received unilateral intra-cerebral injections with a lentiviral (LV)-α-syn vector construct followed by systemic administration of the monoclonal antibody 1H7 (recognizes amino acids 91-99) or control IgG for 3 months. Cerebral α-syn accumulation and axonopathy was assessed by immunohistochemistry and effects on behavior were assessed by Morris water maze. Unilateral LV-α-syn injection resulted in axonal propagation of α-syn in the contra-lateral site with subsequent behavioral deficits and axonal degeneration. Passive immunization with 1H7 antibody reduced the axonal accumulation of α-syn in the contra-lateral side and ameliorated the behavioral deficits. Together this study supports the notion that immunotherapy might improve the deficits in models of synucleinopathy by reducing the axonal propagation and accumulation of α-syn. This represents a potential new mode of action through which α-syn immunization might work.
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Lü, Peng; Yang, Yanhua; Yao, Qin; Xia, Hengchuan; Chen, Keping
2015-01-01
β-thymosin plays important roles in the development of the lymphatic system and the central nervous system in vertebrates. However, its role and function in invertebrates remain much less explored. Here, we firstly isolated a gene encoding β-thymosin in silkworm (Bombyx mori L.). Interestingly, this gene encodes two polypeptides, named as BmTHY1 and BmTHY2, via two different modes of RNA splicing. The recombinant proteins fused with an N-term GST tag were over-expressed in Escherichia coli (E. coli) and further purified to near homogenity to prepare mouse antibodies. The Western blot analysis showed that these proteins were expressed in various tissues and organs, as well as in different developmental stages. Amazingly, the expression of BmTHY2 was hugely increased during the pupae stage, indicating a specialized role in this period. The expression of these proteins was gradually decreased in BmN cells infected by BmNPV, suggesting they may play different roles in the virus infection. In addition, both BmTHY1 and BmTHY2 can interact with 14-3-3 of silkworm and Ubiquitin of BmNPV as shown by GST pull down and Co-IP assays, consistent with their roles in the regulation of the development of nervous system. PMID:26474303
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Mahmoudian, Saeid; Shahmiri, Elaheh; Rouzbahani, Masoumeh; Jafari, Zahra; Keyhani, Mohammad; Rahimi, Farzad; Mahmoudian, Guiti; Akbarvand, Leila; Barzegar, Gholamreza; Farhadi, Mohammad
2011-01-01
Tinnitus is a debilitating condition that is widespread yet difficult to successfully diagnose and treat. This symptom can seriously affect the individual's life quality. The aim of current study was to compose and validate a Persian version of the Tinnitus Handicap Inventory (THI-P). The linguistic validation of the original version of THI into Persian version (THI-P) included translation, back translation and data gathering. The THI-P was administered to 112 tinnitus subjects. Age, gender, medical history and tinnitus characteristics were recorded as baseline information. All participants complained of chronic unilateral or bilateral subjective idiopathic tinnitus lasting for at least 6 months before consulting about their tinnitus. There was no significant difference between gender, age, hearing impairment and total score and subscales of THI-P. Pearson product-moment correlations revealed adequate test-retest reliability for the THI-P (r = 0.96). Cronbach's-alpha coefficient indicated adequate internal stability of the THI-P (r= 0.943), with a total item correction varying between r=0.939 and r=0.944, indicating its reproducibility. The present study proved the internal consistency/ coherency of the Persian version of THI (THI-P). This provides satisfactory application in clinical/research environments.
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Ra, Jin-Ju; Kim, Young Ho
2014-01-01
Background and Objectives The Tinnitus Handicap Inventory (THI) often requires patients to devote their time to complete the questionnaire than they expect. Given that it limits the effectiveness of THI in a busy clinical practice that desires a quick and easy assessment of tinnitus handicap, this study evaluated clinical usefulness of a Simplified version of Tinnitus Handicap Inventory (THI-S) in measuring the severity of tinnitus handicap as well as predicting the psychological distress associated with tinnitus. Subjects and Methods A total of 129 outpatients suffering from tinnitus (61 with normal hearing and 68 with hearing loss) participated in this study. The responses of THI-S (10 items) and THI (25 items) were evaluated to quantify the subjective handicap of tinnitus. The self-perceived level of stress, anxiety, and depression of all participants was measured with a series of self-report questionnaires such as Korean version of Brief Encounter Psychosocial Instrument (BEPSI), State Trait Anxiety Inventory (STAI), and Beck Depression Inventory (BDI), respectively. All the questionnaire responses were analyzed using non-parametric analyses to examine the convergence, comparability, internal consistency reliability and validity of THI-S. Results The severity of tinnitus handicap and the relevant psychological distress greatly varied across individuals. The THI-S responses were comparable to original THI responses, regardless of hearing status of patients. The internal consistency of THI-S responses was found to be good for total score (Cronbach's α=0.83-0.91), with moderately high consistency for the emotional, functional, and catastrophic subscales. Significant (p<0.01) correlations of the THI-S with the THI (rs=0.95) as well as with the BEPSI, STAI, BDI questionnaires (rs=0.40-0.45) suggest that the THI-S questionnaire appeared to be useful to provide objective data of subjective tinnitus handicap as well as predict psychological distress. Three factors were extracted through factor analysis, which explained 73% of the total variance. Conclusions Despite reducing the length of 25 items of THI to 10 items, the THI-S can be used as s a simple but reliable and valid tool for evaluating severity of tinnitus handicap as well as detecting its negative impact on psychological functioning. PMID:24782947
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Preliminary study of synthetic aperture tissue harmonic imaging on in-vivo data
NASA Astrophysics Data System (ADS)
Rasmussen, Joachim H.; Hemmsen, Martin C.; Madsen, Signe S.; Hansen, Peter M.; Nielsen, Michael B.; Jensen, Jørgen A.
2013-03-01
A method for synthetic aperture tissue harmonic imaging is investigated. It combines synthetic aperture sequen- tial beamforming (SASB) with tissue harmonic imaging (THI) to produce an increased and more uniform spatial resolution and improved side lobe reduction compared to conventional B-mode imaging. Synthetic aperture sequential beamforming tissue harmonic imaging (SASB-THI) was implemented on a commercially available BK 2202 Pro Focus UltraView ultrasound system and compared to dynamic receive focused tissue harmonic imag- ing (DRF-THI) in clinical scans. The scan sequence that was implemented on the UltraView system acquires both SASB-THI and DRF-THI simultaneously. Twenty-four simultaneously acquired video sequences of in-vivo abdominal SASB-THI and DRF-THI scans on 3 volunteers of 4 different sections of liver and kidney tissues were created. Videos of the in-vivo scans were presented in double blinded studies to two radiologists for image quality performance scoring. Limitations to the systems transmit stage prevented user defined transmit apodization to be applied. Field II simulations showed that side lobes in SASB could be improved by using Hanning transmit apodization. Results from the image quality study show, that in the current configuration on the UltraView system, where no transmit apodization was applied, SASB-THI and DRF-THI produced equally good images. It is expected that given the use of transmit apodization, SASB-THI could be further improved.
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Shinoda, Kenta; Hirahara, Kiyoshi; Iinuma, Tomohisa; Ichikawa, Tomomi; Suzuki, Akane S.; Sugaya, Kaoru; Tumes, Damon J.; Yamamoto, Heizaburo; Hara, Takahiro; Tani-ichi, Shizue; Ikuta, Koichi; Okamoto, Yoshitaka; Nakayama, Toshinori
2016-01-01
Memory CD4+ T helper (Th) cells are central to long-term protection against pathogens, but they can also be pathogenic and drive chronic inflammatory disorders. How these pathogenic memory Th cells are maintained, particularly at sites of local inflammation, remains unclear. We found that ectopic lymphoid-like structures called inducible bronchus-associated lymphoid tissue (iBALT) are formed during chronic allergic inflammation in the lung, and that memory-type pathogenic Th2 (Tpath2) cells capable of driving allergic inflammation are maintained within the iBALT structures. The maintenance of memory Th2 cells within iBALT is supported by Thy1+IL-7–producing lymphatic endothelial cells (LECs). The Thy1+IL-7–producing LECs express IL-33 and T-cell–attracting chemokines CCL21 and CCL19. Moreover, ectopic lymphoid structures consisting of memory CD4+ T cells and IL-7+IL-33+ LECs were found in nasal polyps of patients with eosinophilic chronic rhinosinusitis. Thus, Thy1+IL-7–producing LECs control chronic allergic airway inflammation by providing a survival niche for memory-type Tpath2 cells. PMID:27140620
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Zhang, F J; Weng, X G; Wang, J F; Zhou, D; Zhang, W; Zhai, C C; Hou, Y X; Zhu, Y H
2014-07-01
Heat stress adversely affects the productivity and immune status of dairy cows. The temperature-humidity index (THI) is commonly used to indicate the degree of heat stress on dairy cattle. We investigated the effects of different THI and Cr supplementation on the antioxidant capacity, the levels of heat shock protein 72 (Hsp72), and cytokine responses of lactating cows. The study used a total of 24 clinically healthy uniparous midlactation Holstein cows, which were randomly divided into 2 groups (n = 12 per group), and was conducted in 3 designated THI periods: low THI period (LTHI; THI = 56.4 ± 2.5), moderate THI period (MTHI; THI = 73.9 ± 1.7), and high THI period (HTHI; THI = 80.3 ± 1.0). The 2 groups of cows were fed corn and corn silage based basal diet supplemented chromium picolinate to provide 3.5 mg of Cr/cow daily (Cr+) or basal diet with no Cr (Cr-). The experiment was a 3 × 2 factorial design. The numbers of leukocytes (P < 0.05) and serum levels of glucose (P < 0.001) were lower; however, the serum levels of blood urea nitrogen (BUN; P < 0.001) and creatinine (P < 0.001) were greater in the MTHI and HTHI than in LTHI. The total antioxidant capacity in the serum was unaltered; an increase in superoxide dismutase activity (P < 0.001) and in serum malondialdehyde concentration (P < 0.001) was observed in the MTHI and HTHI compared with the LTHI. The high THI led to increases in serum concentrations of tumor necrosis factor-α (TNF-α; P < 0.001) and IL-10 (P < 0.05). Cows supplemented with Cr had lower (P = 0.009) serum concentrations of cholesterol but greater (P < 0.001, respectively) serum levels of Hsp72 and IL-10 compared with those without Cr supplementation in the HTHI. Western blot analysis revealed that cows supplemented with Cr had greater (P = 0.038) expression of the inhibitor of nuclear factor kappa B α (IκBα) in peripheral blood mononuclear cells (PBMC) compared with those without Cr supplementation in the HTHI, whereas the expression of Hsp72 in PBMC was unaltered. Data indicate that there is a decrease in glucose and increases in BUN and creatinine in the serum of midlactation cows under hot conditions during the summer and that these cows have a lowered oxidative capacity but an elevated antioxidant capacity. In addition, Cr may play an anti-inflammatory role in lactating cows by promoting the release of Hsp72, increasing the production of IL-10, and inhibiting the degradation of IκBα under hot conditions during the summer.
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DREAM-Dependent Activation of Astrocytes in Amyotrophic Lateral Sclerosis.
Larrodé, Pilar; Calvo, Ana Cristina; Moreno-Martínez, Laura; de la Torre, Miriam; Moreno-García, Leticia; Molina, Nora; Castiella, Tomás; Iñiguez, Cristina; Pascual, Luis Fernando; Mena, Francisco Javier Miana; Zaragoza, Pilar; Y Cajal, Santiago Ramón; Osta, Rosario
2018-01-01
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease of unknown origin and characterized by a relentless loss of motor neurons that causes a progressive muscle weakness until death. Among the several pathogenic mechanisms that have been related to ALS, a dysregulation of calcium-buffering proteins in motor neurons of the brain and spinal cord can make these neurons more vulnerable to disease progression. Downstream regulatory element antagonist modulator (DREAM) is a neuronal calcium-binding protein that plays multiple roles in the nucleus and cytosol. The main aim of this study was focused on the characterization of DREAM and glial fibrillary acid protein (GFAP) in the brain and spinal cord tissues from transgenic SOD1 G93A mice and ALS patients to unravel its potential role under neurodegenerative conditions. The DREAM and GFAP levels in the spinal cord and different brain areas from transgenic SOD1 G93A mice and ALS patients were analyzed by Western blot and immunohistochemistry. Our findings suggest that the calcium-dependent excitotoxicity progressively enhanced in the CNS in ALS could modulate the multifunctional nature of DREAM, strengthening its apoptotic way of action in both motor neurons and astrocytes, which could act as an additional factor to increase neuronal damage. The direct crosstalk between astrocytes and motor neurons can become vulnerable under neurodegenerative conditions, and DREAM could act as an additional switch to enhance motor neuron loss. Together, these findings could pave the way to further study the molecular targets of DREAM to find novel therapeutic strategies to fight ALS.
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Souza, E.L.; Oliveira, C.E.V.; Stamford, T.L.M.; Conceição, M.L.; Neto, N.J. Gomes
2013-01-01
This study evaluated the influence of the phenolic compounds carvacrol (CAR) and thymol (THY) on some physiological characteristics and on the modulation of the secretion of some staphylococcal virulence factors, that is, coagulase and enterotoxin. This study also investigated possible mechanisms for the establishment of the anti-staphylococcal activity of these compounds. Sublethal concentrations (0.3 and 0.15 μL/mL) of CAR and THY inhibited the activity of the enzymes coagulase and lipase and led to a decrease in salt tolerance. At the tested sublethal concentrations, both CAR and THY led to a total suppression of enterotoxin production. The loss of a 260-nm-absorbing material and an efflux of potassium ions occurred immediately after the addition of CAR and THY at 0.6 and 1.2 μL/mL and increased up to 120 min of exposure. Electron microscopy of cells exposed to CAR and THY (0.6 μL/mL) revealed that individual cells appeared to be deformed, with projections of cellular material. The observations of leakage of cellular material and an altered cell surface suggest that gross damage to a cell’s cytoplasmic membrane, which results in a disruption in protein secretion, could be responsible for the anti-staphylococcal properties of CAR and THY. PMID:24159280
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Unraveling Cell Processes: Interference Imaging Interwoven with Data Analysis
Brazhe, A. R.; Pavlov, A. N.; Erokhova, L. A.; Yusipovich, A. I.; Maksimov, G. V.; Mosekilde, E.; Sosnovtseva, O. V.
2006-01-01
The paper presents results on the application of interference microscopy and wavelet-analysis for cell visualization and studies of cell dynamics. We demonstrate that interference imaging of erythrocytes can reveal reorganization of the cytoskeleton and inhomogenity in the distribution of hemoglobin, and that interference imaging of neurons can show intracellular compartmentalization and submembrane structures. We investigate temporal and spatial variations of the refractive index for different cell types: isolated neurons, mast cells and erythrocytes. We show that the refractive dynamical properties differ from cell type to cell type and depend on the cellular compartment. Our results suggest that low frequency variations (0.1–0.6 Hz) result from plasma membrane processes and that higher frequency variations (20–26 Hz) are related to the movement of vesicles. Using double-wavelet analysis, we study the modulation of the 1 Hz rhythm in neurons and reveal its changes under depolarization and hyperpolarization of the plasma membrane. We conclude that interference microscopy combined with wavelet analysis is a useful technique for non-invasive cell studies, cell visualization, and investigation of plasma membrane properties. PMID:19669463
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Vakili, A. R.; Khorrami, B.; Mesgaran, M. Danesh; Parand, E.
2013-01-01
Essential oils have been shown to favorably effect in vitro ruminal fermentation, but there are few in vivo studies that have examined animal responses. The objective of this study was to evaluate the effects of thyme (THY) and cinnamon (CIN) essential oils on feed intake, growth performance, ruminal fermentation and blood metabolites in feedlot calves fed high-concentrate diets. Twelve growing Holstein calves (213±17 kg initial BW) were used in a completely randomized design and received their respective dietary treatments for 45 d. Treatments were: 1-control (no additive), 2-THY (5 g/d/calf) and 3-CIN (5 g/d/calf). Calves were fed ad libitum diets consisting of 15% forage and 85% concentrate, and adapted to the finishing diet by gradually increasing the concentrate ratio with feeding a series of transition diets 5 wk before the experiment started. Supplementation of THY or CIN did not affect DMI and ADG, and feed efficiency was similar between treatment groups. There were no effects of additives on ruminal pH and rumen concentrations of ammonia nitrogen and total VFA; whereas molar proportion of acetate and ratio of acetate to propionate decreased, and the molar proportion of propionate increased with THY and CIN supplementation. Rumen molar concentration of butyrate was significantly increased by adding CIN compared to control; but no change was observed with THY compared with control group. No effects of THY, or CIN were observed on valerate, isobutyrate or isovalerate proportions. Plasma concentrations of glucose, cholesterol, triglyceride, urea-N, β-hydroxybutyrate, alanine aminotransferase and aspartate aminotransferase were not changed by feeding THY or CIN. Results from this study suggest that supplementing a feedlot finishing diet with THY or CIN essential oil might be useful as ruminal fermentation modifiers in beef production systems, but has minor impacts on blood metabolites. PMID:25049871
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Improvement of neuronal differentiation by carbon monoxide: Role of pentose phosphate pathway.
Almeida, Ana S; Soares, Nuno L; Sequeira, Catarina O; Pereira, Sofia A; Sonnewald, Ursula; Vieira, Helena L A
2018-05-15
Over the last decades, the silent-killer carbon monoxide (CO) has been shown to also be an endogenous cytoprotective molecule able to inhibit cell death and modulate mitochondrial metabolism. Neuronal metabolism is mostly oxidative and neurons also use glucose for maintaining their anti-oxidant status by generation of reduced glutathione (GSH) via the pentose-phosphate pathway (PPP). It is established that neuronal differentiation depends on reactive oxygen species (ROS) generation and signalling, however there is a lack of information about modulation of the PPP during adult neurogenesis. Thus, the main goal of this study was to unravel the role of CO on cell metabolism during neuronal differentiation, particularly by targeting PPP flux and GSH levels as anti-oxidant system. A human neuroblastoma SH-S5Y5 cell line was used, which differentiates into post-mitotic neurons by treatment with retinoic acid (RA), supplemented or not with CO-releasing molecule-A1 (CORM-A1). SH-SY5Y cell differentiation supplemented with CORM-A1 prompted an increase in neuronal yield production. It did, however, not alter glycolytic metabolism, but increased the PPP. In fact, CORM-A1 treatment stimulated (i) mRNA expression of 6-phosphogluconate dehydrogenase (PGDH) and transketolase (TKT), which are enzymes for oxidative and non-oxidative phases of the PPP, respectively and (ii) protein expression and activity of glucose 6-phosphate dehydrogenase (G6PD) the rate-limiting enzyme of the PPP. Likewise, whenever G6PD was knocked-down CO-induced improvement on neuronal differentiation was reverted, while pharmacological inhibition of GSH synthesis did not change CO's effect on the improvement of neuronal differentiation. Both results indicate the key role of PPP in CO-modulation of neuronal differentiation. Furthermore, at the end of SH-SY5Y neuronal differentiation process, CORM-A1 supplementation increased the ratio of reduced and oxidized glutathione (GSH/GSSG) without alteration of GSH metabolism. These data corroborate with PPP stimulation. In conclusion, CO improves neuronal differentiation of SH-S5Y5 cells by stimulating the PPP and modulating the GSH system. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.
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Berman, A; Horovitz, Talia; Kaim, M; Gacitua, H
2016-10-01
The combined temperature-humidity heat stress is estimated in farm animals by indices derived of an index based on human thermal comfort sensation. The latter index consists of temperature and humidity measures that sum to form the temperature-humidity index (THI). The hitherto unknown relative contribution of temperature and humidity to the THI was examined. A temperature-humidity data set (temperature 20-42 °C and relative humidity 10-70 %) was used to assess by regression procedures the relative weights of temperature and humidity in the variance of THI values produced by six commonly used heat stress indices. The temperature (Ta) effect was predominant (0.82-0.95 of variance) and humidity accounted for only 0.05 to 0.12 of THI variance, half of the variance encountered in animal responses to variable humidity heat stress. Significant difference in THI values was found between indices in the relative weights of temperature and humidity. As in THI indices, temperature and humidity are expressed in different physical units, their sum has no physical attributes, and empirical evaluations assess THI relation to animal responses. A sensible heat THI was created, in which at higher temperatures humidity reaches 0.25 of sensible heat, similarly to evaporative heat loss span in heat stressed animals. It relates to ambient temperature-humidity similarly to present THI; its values are similar to other THI but greater at higher humidity. In warm conditions, mean animal responses are similar in both indices. The higher sensitivity to humidity makes this index preferable for warm-humid conditions.
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Psychometric Properties of the Persian Version of the Tinnitus Handicap Inventory (THI-P)
Jalali, Mir Mohammad; Soleimani, Robabeh; Fallahi, Mahnaz; Aghajanpour, Mohammad; Elahi, Masoumeh
2015-01-01
Introduction: Tinnitus can have a significant effect on an individual’s quality of life, and is very difficult quantify. One of the most popular questionnaires used in this area is the Tinnitus Handicap Inventory (THI). The aim of this study was to determine the reliability and validity of a Persian translation of the Tinnitus Handicap Inventory (THI-P). Materials and Methods: This prospective clinical study was performed in the Otolaryngology Department of Guilan University of Medical Sciences, Iran. A total of 102 patients aged 23–80 years with tinnitus completed the (THI-P). The patients were instructed to complete the Beck Depression Inventory (BDI) and the State-Trait Anxiety Inventory (STAI). Audiometry was performed. Eight-five patients were asked to complete the THI-P for a second time 7–10 days after the initial interview. We assessed test–retest reliability and internal reliability of the THI-P. Validity was assessed by analyzing the THI-P of patients according to their age, tinnitus duration and psychological distress (BDI and STAI). A factor analysis was computed to verify if three subscales (functional, emotional, and catastrophic) represented three distinct variables. Results: Test–retest correlation coefficient scores were highly significant. The THI-P and its subscales showed good internal consistency reliability (α = 0.80 to 0.96). High-to-moderate correlations were observed between THI-P and psychological distress and tinnitus symptom ratings. A confirmatory factor analysis failed to validate the three subscales of THI, and high inter-correlations found between the subscales question whether they represent three distinct factors. Conclusion: The results suggest that the THI-P is a reliable and valid tool which can be used in a clinical setting to quantify the impact of tinnitus on the quality of life of Iranian patients. PMID:25938079
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Enrichment and isolation of neurons from adult mouse brain for ex vivo analysis.
Berl, Sabina; Karram, Khalad; Scheller, Anja; Jungblut, Melanie; Kirchhoff, Frank; Waisman, Ari
2017-05-01
Isolation of neurons from the adult mouse CNS is important in order to study their gene expression during development or the course of different diseases. Here we present two different methods for the enrichment or isolation of neurons from adult mouse CNS. These methods: are either based on flow cytometry sorting of eYFP expressing neurons, or by depletion of non-neuronal cells by sorting with magnetic-beads. Enrichment by FACS sorting of eYFP positive neurons results in a population of 62.4% NeuN positive living neurons. qPCR data shows a 3-5fold upregulation of neuronal markers. The isolation of neurons based on depletion of non-neuronal cells using the Miltenyi Neuron Isolation Kit, reaches a purity of up to 86.5%. qPCR data of these isolated neurons shows an increase in neuronal markers and an absence of glial markers, proving pure neuronal RNA isolation. Former data related to neuronal gene expression are mainly based on histology, which does not allow for high-throughput transcriptome analysis to examine differential gene expression. These protocols can be used to study cell type specific gene expression of neurons to unravel their function in the process of damage to the CNS. Copyright © 2017 Elsevier B.V. All rights reserved.
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Honeycutt, Jenna B; Wahl, Angela; Archin, Nancie; Choudhary, Shailesh; Margolis, David; Garcia, J Victor
2013-10-24
The major targets of HIV infection in humans are CD4⁺ T cells. CD4⁺ T cell depletion is a hallmark of AIDS. Previously, the SCID-hu thy/liv model was used to study the effect of HIV on thymopoeisis in vivo. However, these mice did not develop high levels of peripheral T cell reconstitution and required invasive surgery for infection and analysis. Here, we describe a novel variant of this model in which thy/liv implantation results in systemic reconstitution with human T cells in the absence of any other human hematopoietic lineages. NOD/SCID-hu thy/liv and NSG-hu thy/liv mice were created by implanting human fetal thymus and liver tissues under the kidney capsule of either NOD/SCID or NSG mice. In contrast to NOD/SCID-hu thy/liv mice that show little or no human cells in peripheral blood or tissues, substantial systemic human reconstitution occurs in NSG-hu thy/liv. These mice are exclusively reconstituted with human T cells (i.e. T-cell only mice or TOM). Despite substantial levels of human T cells no signs of graft-versus-host disease (GVHD) were noted in these mice over a period of 14 months. TOM are readily infected after parenteral exposure to HIV-1. HIV replication is sustained in peripheral blood at high levels and results in modest reduction of CD4⁺ T cells. HIV-1 replication in TOM responds to daily administration of combination antiretroviral therapy (ART) resulting in strong suppression of virus replication as determined by undetectable viral load in plasma. Latently HIV infected resting CD4⁺ T cells can be isolated from suppressed mice that can be induced to express HIV ex-vivo upon activation demonstrating the establishment of latency in vivo. NSG-hu thy/liv mice are systemically reconstituted with human T cells. No other human lymphoid lineages are present in these mice (i.e. monocytes/macrophages, B cells and DC are all absent). These T cell only mice do not develop GVHD, are susceptible to HIV-1 infection and can efficiently maintain virus replication. HIV infected TOM undergoing ART harbor latently infected, resting CD4+ T cells.
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Huang, Kaipeng; Li, Ruiming; Wei, Wentao
2018-08-05
Mesangial proliferative glomerulonephritis (MsPGN) is characterized by glomerular mesangial cells proliferation and extracellular matrix deposition in mesangial area, which develop into glomerulosclerosis. Both silent information regulator 2-related protein 1 (Sirt1) and nuclear factor erythroid 2-related factor 2/anti-oxidant response element (Nrf2/ARE) pathway had remarkable renoprotective effects. However, whether Sirt1 and Nrf2/ARE pathway can regulate the pathological process of MsPGN remains unknown. Here, we found that Sirt1 activation by SRT1720 decreased mesangial hypercellularity and mesangial matrix areas, reduced renal Col4 and α-SMA expressions, lowered 24 h proteinuria, and eventually reduced FN and TGF-β1 expressions in rats received anti-Thy 1.1 IgG. Further study showed that SRT1720 markedly enhanced the activity of Nrf2/ARE pathway including promoting the nuclear content and ARE-binding ability of Nrf2, elevating the protein levels of HO-1 and SOD1, two target genes of Nrf2, which eventually increased total SOD activity and decreased malondialdehyde level in the kidney tissues of experimental anti-Thy 1.1 MsPGN rats. Taken together, Sirt1 prevented the pathological process of experimental anti-Thy 1.1 MsPGN through promoting the activation of Nrf2/ARE pathway, which warrants further elucidation. Sirt1 might be a potential therapeutic target for treating MsPGN. Copyright © 2018 Elsevier B.V. All rights reserved.
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Shu, Sai-Nan; Wei, Lai; Wang, Jiang-Hua; Zhan, Yu-Tao; Chen, Hong-Song; Wang, Yu
2004-10-01
To investigate the different effects of mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) on hepatic differentiation. MSCs from rat bone marrow were isolated and cultured by standard methods. HSCs from rat bone marrow were isolated and purified by magnetic activated cell sorting. Both cell subsets were induced. Morphology, RT-PCR and immunocytochemistry were used to identify the hepatic differentiation grade. MSCs exhibited round in shape after differentiation, instead of fibroblast-like morphology before differentiation. Albumin mRNA and protein were expressed positively in MSCs, without detection of alpha-fetoprotein (AFP). HSCs were polygonal in shape after differentiation. The expression of albumin signal decreased and AFP signal increased. The expression of CK18 was continuous in MSCs and HSCs both before and after induction. Both MSCs and HSCs have hepatic differentiation capabilities. However, their capabilities are not the same. MSCs can differentiate into mature hepatocyte-like cells, never expressing early hepatic specific genes, while Thy-1.1(+) cells are inclined to differentiate into hepatic stem cell-like cells, with an increasing AFP expression and a decreasing albumin signal. CK18 mRNA is positive in Thy-1.1(+) cells and MSCs, negative in Thy-1.1(-) cells. It seems that CK18 has some relationship with Thy-1.1 antigen, and CK18 may be a predictive marker of hepatic differentiation capability.
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Games, Dora; Seubert, Peter; Rockenstein, Edward; Patrick, Christina; Trejo, Margarita; Ubhi, Kiren; Ettle, Benjamin; Ghassemiam, Majid; Barbour, Robin; Schenk, Dale; Nuber, Silke; Masliah, Eliezer
2014-01-01
Progressive accumulation of α-synuclein (α-syn) in limbic and striatonigral systems is associated with the neurodegenerative processes in dementia with Lewy bodies (DLB) and Parkinson’s disease (PD). The murine Thy-1 (mThy1)-α-syn transgenic (tg) model recapitulates aspects of degenerative processes associated with α-syn accumulation in these disorders. Given that axonal and synaptic pathologies are important features of DLB and PD, we sought to investigate the extent and characteristics of these alterations in mThy1-α-syn tg mice and to determine the contribution of α-syn c-terminally cleaved at amino acid 122 (CT α-syn) to these abnormalities. We generated a novel polyclonal antibody (SYN105) against the c-terminally truncated sequence (amino acids 121 to 123) of α-syn (CT α-syn) and performed immunocytochemical and ultrastructural analyses in mThy1-α-syn tg mice. We found abundant clusters of dystrophic neurites in layers 2 to 3 of the neocortex, the stratum lacunosum, the dentate gyrus, and cornu ammonis 3 of the hippocampus, striatum, thalamus, midbrain, and pons. Dystrophic neurites displayed intense immunoreactivity detected with the SYN105 antibody. Double-labeling studies with antibodies to phosphorylated neurofilaments confirmed the axonal location of full-length and CT α-syn. α-Syn immunoreactive dystrophic neurites contained numerous electrodense laminated structures. These results show that neuritic dystrophy is a prominent pathologic feature of the mThy1-α-syn tg model and suggest that CT α-syn might play an important role in the process of axonal damage in these mice as well as in DLB and PD. PMID:23313024
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Games, Dora; Seubert, Peter; Rockenstein, Edward; Patrick, Christina; Trejo, Margarita; Ubhi, Kiren; Ettle, Benjamin; Ghassemiam, Majid; Barbour, Robin; Schenk, Dale; Nuber, Silke; Masliah, Eliezer
2013-03-01
Progressive accumulation of α-synuclein (α-syn) in limbic and striatonigral systems is associated with the neurodegenerative processes in dementia with Lewy bodies (DLB) and Parkinson's disease (PD). The murine Thy-1 (mThy1)-α-syn transgenic (tg) model recapitulates aspects of degenerative processes associated with α-syn accumulation in these disorders. Given that axonal and synaptic pathologies are important features of DLB and PD, we sought to investigate the extent and characteristics of these alterations in mThy1-α-syn tg mice and to determine the contribution of α-syn c-terminally cleaved at amino acid 122 (CT α-syn) to these abnormalities. We generated a novel polyclonal antibody (SYN105) against the c-terminally truncated sequence (amino acids 121 to 123) of α-syn (CT α-syn) and performed immunocytochemical and ultrastructural analyses in mThy1-α-syn tg mice. We found abundant clusters of dystrophic neurites in layers 2 to 3 of the neocortex, the stratum lacunosum, the dentate gyrus, and cornu ammonis 3 of the hippocampus, striatum, thalamus, midbrain, and pons. Dystrophic neurites displayed intense immunoreactivity detected with the SYN105 antibody. Double-labeling studies with antibodies to phosphorylated neurofilaments confirmed the axonal location of full-length and CT α-syn. α-Syn immunoreactive dystrophic neurites contained numerous electrodense laminated structures. These results show that neuritic dystrophy is a prominent pathologic feature of the mThy1-α-syn tg model and suggest that CT α-syn might play an important role in the process of axonal damage in these mice as well as in DLB and PD. Copyright © 2013 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.
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Morignat, Eric; Gay, Emilie; Vinard, Jean-Luc; Calavas, Didier; Hénaux, Viviane
2015-07-01
In the context of climate change, the frequency and severity of extreme weather events are expected to increase in temperate regions, and potentially have a severe impact on farmed cattle through production losses or deaths. In this study, we used distributed lag non-linear models to describe and quantify the relationship between a temperature-humidity index (THI) and cattle mortality in 12 areas in France. THI incorporates the effects of both temperature and relative humidity and was already used to quantify the degree of heat stress on dairy cattle because it does reflect physical stress deriving from extreme conditions better than air temperature alone. Relationships between daily THI and mortality were modeled separately for dairy and beef cattle during the 2003-2006 period. Our general approach was to first determine the shape of the THI-mortality relationship in each area by modeling THI with natural cubic splines. We then modeled each relationship assuming a three-piecewise linear function, to estimate the critical cold and heat THI thresholds, for each area, delimiting the thermoneutral zone (i.e. where the risk of death is at its minimum), and the cold and heat effects below and above these thresholds, respectively. Area-specific estimates of the cold or heat effects were then combined in a hierarchical Bayesian model to compute the pooled effects of THI increase or decrease on dairy and beef cattle mortality. A U-shaped relationship, indicating a mortality increase below the cold threshold and above the heat threshold was found in most of the study areas for dairy and beef cattle. The pooled estimate of the mortality risk associated with a 1°C decrease in THI below the cold threshold was 5.0% for dairy cattle [95% posterior interval: 4.4, 5.5] and 4.4% for beef cattle [2.0, 6.5]. The pooled mortality risk associated with a 1°C increase above the hot threshold was estimated to be 5.6% [5.0, 6.2] for dairy and 4.6% [0.9, 8.7] for beef cattle. Knowing the thermoneutral zone and temperature effects outside this zone is of primary interest for farmers because it can help determine when to implement appropriate preventive and mitigation measures. Copyright © 2015 Elsevier Inc. All rights reserved.
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Oxidizer heat exchanger component test
NASA Technical Reports Server (NTRS)
Kanic, P. G.
1988-01-01
The RL10-IIB engine, is capable of multimode thrust operation. The engine operates at two low-thrust levels: tank head idle (THI), approximately 1 to 2 percent of full thrust; and pumped idle, 10 percent of full thrust. Operation at THI provides vehicle propellant settling thrust and efficient thermal conditioning; PI operation provides vehicle tank prepressurization and maneuver thrust for low-g deployment. Stable combustion of the RL10-IIB engine during the low-thrust operating modes can be accomplished by using a heat exchanger to supply gaseous oxygen to the propellant injector. The oxidized heat exchanger (OHE) vaporizes the liquid oxygen using hydrogen as the energy source. This report summarizes the test activity and post-test data analysis for two possible heat exchangers, each of which employs a completely different design philosophy. One design makes use of a low-heat transfer (PHT) approach in combination with a volume to attenuate pressure and flow oscillations. The test data showed that the LHT unit satisfied the oxygen exit quality of 0.95 or greater in both the THI and PI modes while maintaining stability. The HHT unit fulfilled all PI requirements; data for THI satisfactory operation is implied from experimental data that straddle the exact THI operating point.
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Sadakane, Chiharu; Kase, Yoshio; Koseki, Junichi; Hasegawa, Yoshihiro; Shindo, Shoichiro; Maruyama, Hirobumi; Takeda, Shuichi; Takeda, Hiroshi; Hattori, Tomohisa
2011-02-01
Phosphodiesterase type IV (PDEIV) plays an important role in the immune response and inflammation. However, it is well known that classical PDEIV inhibitors have systemic side effects, so the clinical and chronic use of these agents as therapy for glomerulonephritis is difficult. This study was performed to elucidate the anti-nephritic effects of TJN-598, a new chemical compound derived from herbal components, on experimental mesangial proliferative glomerulonephritis. We first examined the effects of TJN-598 and captopril on mesangial expansion induced by anti-Thy1 serum in rats. Second, to investigate the effects of TJN-598 and rolipram, which are typical PDEIV inhibitors, on the production of tumor necrosis factor (TNF)-α and transforming growth factor (TGF)-β1, glomeruli were isolated from rats with anti-Thy1 nephritis and incubated with the test drugs in vitro for 48 h. Treatment with TJN-598 prevented an increase in the mesangial area/total glomerular area, in the number of cells in the glomerular cross section and matrix index. TJN-598 also inhibited the increases in the expression of α-smooth muscle actin, the TGF-β1-positive area, in the number of ED-1 positive cells and proliferating cell nuclear antigen-positive cells in the glomeruli. Furthermore, administration of TJN-598 inhibited increases in the levels of TGF-β1 protein derived from glomeruli with anti-Thy-1 nephritis. The addition of both TJN-598 and rolipram to the culture supernatant inhibited both increased expression of TGF-β1 and increases in levels of TNF-α in glomeruli isolated from rats with anti-Thy1 nephritis in a dose-dependent manner. These results suggest that TJN-598, a PDEIV inhibitor, is effective against expansion of mesangial cells, via the suppression of secretion of TGF-β1 and TNF-α from inflamed glomeruli.
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Effectiveness of a tinnitus management programme: a 2-year follow-up study
Gudex, Claire; Skellgaard, Preben H; West, Torben; Sørensen, Jan
2009-01-01
Background Tinnitus impairs the possibility of leading a normal life in 0.5–1% of the population. While neither medical nor surgical treatment appears effective, counselling may offer some relief. An intervention combining counselling and hearing devices is offered to clients referred to the Centre for Help Aids and Communication (CHC) in southern Denmark. The aims of this exploratory study were to examine i) the characteristics of CHC's clients and their tinnitus, ii) the effectiveness of the treatment, and iii) whether particular client groups benefit more than others. Methods One hundred new clients presenting with tinnitus completed the Tinnitus Handicap Inventory (THI) three times – before their first consultation, after one month and after 1–2 years. The scores were tested for significant differences over time using tests for paired data. Logistic regression was used to examine factors associated with a clinically important difference (i.e. THI score improvement of at least 20 points). Results At final follow-up, total THI score was significantly lower than baseline, i.e. 29.8 (CI 25.5–34.2) vs. 37.2 (CI 33.1–37.2), p < 0.01. The programme achieved a clinically important difference for 27% and 24% of the clients one month and 1–2 years after the first consultation, respectively. It appeared that greater improvement in THI score was related to higher baseline THI score and possibly also to treatment by a particular CHC therapist. The absolute reduction in mean THI score after 1–2 years for clients with moderate and severe handicap was 14 and 20 points, respectively, i.e. similar to that previously reported for TRT (14–28 points). The cost of the current programme was approximately 200 EUR per client. Conclusion The tinnitus management programme appeared to provide significant benefit to many clients at a relatively low cost. It would be useful to conduct a randomised controlled study comparing the current programme with alternative forms of combination counselling/sound therapy approaches. PMID:19558680
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The effects of heat stress in Italian Holstein dairy cattle.
Bernabucci, U; Biffani, S; Buggiotti, L; Vitali, A; Lacetera, N; Nardone, A
2014-01-01
The data set for this study comprised 1,488,474 test-day records for milk, fat, and protein yields and fat and protein percentages from 191,012 first-, second-, and third-parity Holstein cows from 484 farms. Data were collected from 2001 through 2007 and merged with meteorological data from 35 weather stations. A linear model (M1) was used to estimate the effects of the temperature-humidity index (THI) on production traits. Least squares means from M1 were used to detect the THI thresholds for milk production in all parities by using a 2-phase linear regression procedure (M2). A multiple-trait repeatability test-model (M3) was used to estimate variance components for all traits and a dummy regression variable (t) was defined to estimate the production decline caused by heat stress. Additionally, the estimated variance components and M3 were used to estimate traditional and heat-tolerance breeding values (estimated breeding values, EBV) for milk yield and protein percentages at parity 1. An analysis of data (M2) indicated that the daily THI at which milk production started to decline for the 3 parities and traits ranged from 65 to 76. These THI values can be achieved with different temperature/humidity combinations with a range of temperatures from 21 to 36°C and relative humidity values from 5 to 95%. The highest negative effect of THI was observed 4 d before test day over the 3 parities for all traits. The negative effect of THI on production traits indicates that first-parity cows are less sensitive to heat stress than multiparous cows. Over the parities, the general additive genetic variance decreased for protein content and increased for milk yield and fat and protein yield. Additive genetic variance for heat tolerance showed an increase from the first to third parity for milk, protein, and fat yield, and for protein percentage. Genetic correlations between general and heat stress effects were all unfavorable (from -0.24 to -0.56). Three EBV per trait were calculated for each cow and bull (traditional EBV, traditional EBV estimated with the inclusion of THI covariate effect, and heat tolerance EBV) and the rankings of EBV for 283 bulls born after 1985 with at least 50 daughters were compared. When THI was included in the model, the ranking for 17 and 32 bulls changed for milk yield and protein percentage, respectively. The heat tolerance genetic component is not negligible, suggesting that heat tolerance selection should be included in the selection objectives. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
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Manipulating neuronal activity with low frequency transcranial ultrasound
NASA Astrophysics Data System (ADS)
Moore, Michele Elizabeth
Stimulation of the rodent cerebral cortex is used to investigate the underlying biological basis for the restorative effects of slow wave sleep. Neuronal activation by optogenetic and ultrasound stimulation elicits changes in action potentials across the cerebral cortex that are recorded as electroencephalograms. Optogenetic stimulation requires an invasive implantation procedure limiting its application in human studies. We sought to determine whether ultrasound stimulation could be as effective as optogenetic techniques currently used, in an effort to further understand the physiological and metabolic requirements of sleep. We successfully recorded electroencephalograms in response to transcranial ultrasound stimulation of the barrel cortex at 1 and 7 Hz frequencies, comparing them to those recorded in response to optogenetic stimuli applied at the same frequencies. Our results showed application of a 473 nm blue LED positioned 6 cm above the skull and ultrasound stimulation at an output voltage of 1000 mVpp produced electroencephalograms with physiological responses of similar amplitude. We concluded that there exists an intensity-proportionate response in the optogenetic stimulation, but not with ultrasound stimulation at the frequencies we surveyed. Activation of neuronal cells in response to optogenetic stimulation in a Thy1-ChR2 transgenic mouse line is specifically targeted to pyramidal cells in the cerebral cortex. ChR2 responses to optogenetic stimulation are mediated by a focal activation of neuronal ion channels. We measured electrophysiological responses to ultrasound stimulation, comparing them to those recorded from optogenetic stimuli. Our results show striking similarities between ultrasound-induced responses and optogenetically-induced responses, which may indicate that transcranial ultrasound stimulation is also mediated by ion channel dependent processes in cerebral cortical neurons. The biophysical substrates for electrical excitability of neurons impose temporal constraints on their response to stimulation. If ultrasound-mediated responses are, in fact, ion channel mediated responses, ultrasound-induced responses should exhibit time-dependence characteristics similar to those of optogenetically-triggered responses. Minimal stimulus duration thresholds and the temporal limits of paired pulse facilitation for ultrasound stimulation were identical to those of optogenetic stimulation. Collectively, these experiments demonstrate an electrophysiological basis for low-frequency transcranial ultrasound stimulation of cerebral cortical neuronal activity.
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Wagener, Kerstin C; Kolbrink, Benedikt; Dietrich, Katharina; Kizina, Kathrin M; Terwitte, Lukas S; Kempkes, Belinda; Bao, Guobin; Müller, Michael
2016-07-01
Reactive oxygen species (ROS) and downstream redox alterations not only mediate physiological signaling but also neuropathology. For long, ROS/redox imaging was hampered by a lack of reliable probes. Genetically encoded redox sensors overcame this gap and revolutionized (sub)cellular redox imaging. Yet, the successful delivery of sensor-coding DNA, which demands transfection/transduction of cultured preparations or stereotaxic microinjections of each subject, remains challenging. By generating transgenic mice, we aimed to overcome limiting cultured preparations, circumvent surgical interventions, and to extend effectively redox imaging to complex and adult preparations. Our redox indicator mice widely express Thy1-driven roGFP1 (reduction-oxidation-sensitive green fluorescent protein 1) in neuronal cytosol or mitochondria. Negative phenotypic effects of roGFP1 were excluded and its proper targeting and functionality confirmed. Redox mapping by ratiometric wide-field imaging reveals most oxidizing conditions in CA3 neurons. Furthermore, mitochondria are more oxidized than cytosol. Cytosolic and mitochondrial roGFP1s reliably report cell endogenous redox dynamics upon metabolic challenge or stimulation. Fluorescence lifetime imaging yields stable, but marginal, response ranges. We therefore developed automated excitation ratiometric 2-photon imaging. It offers superior sensitivity, spatial resolution, and response dynamics. Redox indicator mice enable quantitative analyses of subcellular redox dynamics in a multitude of preparations and at all postnatal stages. This will uncover cell- and compartment-specific cerebral redox signals and their defined alterations during development, maturation, and aging. Cross-breeding with other disease models will reveal molecular details on compartmental redox homeostasis in neuropathology. Combined with ratiometric 2-photon imaging, this will foster our mechanistic understanding of cellular redox signals in their full complexity. Antioxid. Redox Signal. 25, 41-58.
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Wagener, Kerstin C.; Kolbrink, Benedikt; Dietrich, Katharina; Kizina, Kathrin M.; Terwitte, Lukas S.; Kempkes, Belinda; Bao, Guobin
2016-01-01
Abstract Aims: Reactive oxygen species (ROS) and downstream redox alterations not only mediate physiological signaling but also neuropathology. For long, ROS/redox imaging was hampered by a lack of reliable probes. Genetically encoded redox sensors overcame this gap and revolutionized (sub)cellular redox imaging. Yet, the successful delivery of sensor-coding DNA, which demands transfection/transduction of cultured preparations or stereotaxic microinjections of each subject, remains challenging. By generating transgenic mice, we aimed to overcome limiting cultured preparations, circumvent surgical interventions, and to extend effectively redox imaging to complex and adult preparations. Results: Our redox indicator mice widely express Thy1-driven roGFP1 (reduction–oxidation-sensitive green fluorescent protein 1) in neuronal cytosol or mitochondria. Negative phenotypic effects of roGFP1 were excluded and its proper targeting and functionality confirmed. Redox mapping by ratiometric wide-field imaging reveals most oxidizing conditions in CA3 neurons. Furthermore, mitochondria are more oxidized than cytosol. Cytosolic and mitochondrial roGFP1s reliably report cell endogenous redox dynamics upon metabolic challenge or stimulation. Fluorescence lifetime imaging yields stable, but marginal, response ranges. We therefore developed automated excitation ratiometric 2-photon imaging. It offers superior sensitivity, spatial resolution, and response dynamics. Innovation and Conclusion: Redox indicator mice enable quantitative analyses of subcellular redox dynamics in a multitude of preparations and at all postnatal stages. This will uncover cell- and compartment-specific cerebral redox signals and their defined alterations during development, maturation, and aging. Cross-breeding with other disease models will reveal molecular details on compartmental redox homeostasis in neuropathology. Combined with ratiometric 2-photon imaging, this will foster our mechanistic understanding of cellular redox signals in their full complexity. Antioxid. Redox Signal. 25, 41–58. PMID:27059697
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Temperature humidity index scenarios in the Mediterranean basin.
Segnalini, M; Bernabucci, U; Vitali, A; Nardone, A; Lacetera, N
2013-05-01
The study was undertaken to describe the temperature humidity index (THI) dynamics over the Mediterranean basin for the period 1971-2050. The THI combines temperature and humidity into a single value, and has been widely used to predict the effects of environmental warmth in farm animals. The analysis was based on daily outputs of the temperature and relative humidity from the Max Planck Institute data using the Intergovernmental Panel on Climate Change Special Report Emission Scenario A1B. Data revealed a gradual increase of both annual and seasonal THI during the period under investigation and a strong heterogeneity of the Mediterranean area. In particular, the analysis indicated that Spain, southern France and Italy should be expected to undergo the highest THI increase, which in the last decade under study (2041-2050) will range between 3 and 4 units. However, only during summer months the area presents characteristics indicating risk of thermal (heat) stress for farm animals. In this regard, scenario maps relative to the summer season suggested an enlargement of the areas in the basin where summer THI values will likely cause thermal discomfort in farm animals. In conclusion, the study indicated that the Mediterranean basin is likely to undergo THI changes, which may aggravate the consequences of hot weather on animal welfare, performances, health and survival and may help farmers, nutritionists, veterinarians, and policy-makers to develop appropriate adaptation strategies to limit consequences of climate change for the livestock sector in the Mediterranean countries.
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Temperature humidity index scenarios in the Mediterranean basin
NASA Astrophysics Data System (ADS)
Segnalini, M.; Bernabucci, U.; Vitali, A.; Nardone, A.; Lacetera, N.
2013-05-01
The study was undertaken to describe the temperature humidity index (THI) dynamics over the Mediterranean basin for the period 1971-2050. The THI combines temperature and humidity into a single value, and has been widely used to predict the effects of environmental warmth in farm animals. The analysis was based on daily outputs of the temperature and relative humidity from the Max Planck Institute data using the Intergovernmental Panel on Climate Change Special Report Emission Scenario A1B. Data revealed a gradual increase of both annual and seasonal THI during the period under investigation and a strong heterogeneity of the Mediterranean area. In particular, the analysis indicated that Spain, southern France and Italy should be expected to undergo the highest THI increase, which in the last decade under study (2041-2050) will range between 3 and 4 units. However, only during summer months the area presents characteristics indicating risk of thermal (heat) stress for farm animals. In this regard, scenario maps relative to the summer season suggested an enlargement of the areas in the basin where summer THI values will likely cause thermal discomfort in farm animals. In conclusion, the study indicated that the Mediterranean basin is likely to undergo THI changes, which may aggravate the consequences of hot weather on animal welfare, performances, health and survival and may help farmers, nutritionists, veterinarians, and policy-makers to develop appropriate adaptation strategies to limit consequences of climate change for the livestock sector in the Mediterranean countries.
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Krashes, Michael J.; Shah, Bhavik P.; Koda, Shuichi; Lowell, Bradford B.
2013-01-01
Summary Agouti-related peptide (AgRP) neurons of the hypothalamus release a fast transmitter (GABA) in addition to neuropeptides (NPY and AgRP). This raises questions as to their respective functions. Acute activation of AgRP neurons robustly promotes food intake, while central injections of AgRP, NPY or GABA agonist results in marked escalation of food consumption with temporal variance. Given the orexigenic capability of all three of these neuroactive substances in conjunction with their coexpression in AgRP neurons, we looked to unravel their relative temporal role in driving food intake. Following acute stimulation of AgRP neurons using DREADD technology, we found that either GABA or NPY is required for rapid stimulation of feeding, and the neuropeptide AgRP, through action on MC4 receptors, is sufficient to induce feeding over a delayed, yet prolonged period. These studies help to elucidate the neurochemical mechanisms of AgRP neurons in controlling temporally distinct phases of eating. PMID:24093681
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Anxiety-like behavior in transgenic mice with brain expression of neuropeptide Y.
Inui, A; Okita, M; Nakajima, M; Momose, K; Ueno, N; Teranishi, A; Miura, M; Hirosue, Y; Sano, K; Sato, M; Watanabe, M; Sakai, T; Watanabe, T; Ishida, K; Silver, J; Baba, S; Kasuga, M
1998-01-01
Neuropeptide Y (NPY), one of the most abundant peptide transmitters in the mammalian brain, is assumed to play an important role in behavior and its disorders. To understand the long-term modulation of neuronal functions by NPY, we raised transgenic mice created with a novel central nervous system (CNS) neuron-specific expression vector of human Thy- gene fragment linked to mouse NPY cDNA. In situ hybridization analysis demonstrated transgene-derived NPY expression in neurons (e.g., in the hippocampus, cerebral cortex, and the arcuate nucleus of the hypothalamus) in the transgenic mice. The modest increase of NPY protein in the brain was demonstrated by semiquantitative immunohistochemical analysis and by radioreceptor assay (115% in transgenic mice compared to control littermates). Double-staining experiments indicated colocalization of the transgene-derived NPY message and NPY protein in the same neurons, such as in the arcuate nucleus. The transgenic mice displayed behavioral signs of anxiety and hypertrophy of adrenal zona fasciculata cells, but no change in food intake was observed. The anxiety-like behavior of transgenic mice was reversed, at least in part, by administration of corticotropin-releasing factor (CRF) antagonists, alpha-helical CRF9-41, into the third cerebral ventricle. These results suggest that NPY has a role in anxiety and behavioral responses to stress partly via the CRF neuronal system. This genetic model may provide a unique opportunity to study human anxiety and emotional disorders.
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Unal-Cevik, Isin; Gursoy-Ozdemir, Yasemin; Yemisci, Muge; Lule, Sevda; Gurer, Gunfer; Can, Alp; Müller, Veronica; Kahle, Philip J; Dalkara, Turgay
2011-01-01
Alpha-synuclein oligomerization and aggregation are considered to have a role in the pathogenesis of neurodegenerative diseases. However, despite numerous in vitro studies, the impact of aggregates in the intact brain is unclear. In vitro, oxidative/nitrative stress and acidity induce α-synuclein oligomerization. These conditions favoring α-synuclein fibrillization are present in the ischemic brain, which may serve as an in vivo model to study α-synuclein aggregation. In this study, we show that 30-minute proximal middle cerebral artery (MCA) occlusion and 72 hours reperfusion induce oligomerization of wild-type α-synuclein in the ischemic mouse brain. The nonamyloidogenic isoform β-synuclein did not form oligomers. Alpha-synuclein aggregates were confined to neurons and colocalized with ubiquitin immunoreactivity. We also found that 30 minutes proximal MCA occlusion and 24 hours reperfusion induced larger infarcts in C57BL/6(Thy1)-h[A30P]alphaSYN transgenic mice, which have an increased tendency to form synuclein fibrils. Trangenics also developed more selective neuronal necrosis when subjected to 20 minutes distal MCA occlusion and 72 hours reperfusion. Enhanced 3-nitrotyrosine immunoreactivity in transgenic mice suggests that oxidative/nitrative stress may be one of the mechanisms mediating aggregate toxicity. Thus, the increased vulnerability of transgenic mice to ischemia suggests that α-synuclein aggregates not only form during ischemia but also negatively impact neuronal survival, supporting the idea that α-synuclein misfolding may be neurotoxic. PMID:20877387
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Shankar, Akshaya; Jagota, Anand; Mittal, Jeetain
2012-10-11
Single- and double-stranded DNA are increasingly being paired with surfaces and nanoparticles for numerous applications, such as sensing, imaging, and drug delivery. Unlike the majority of DNA structures in bulk that are stabilized by canonical Watson-Crick pairing between Ade-Thy and Gua-Cyt, those adsorbed on surfaces are often stabilized by noncanonical base pairing, quartet formation, and base-surface stacking. Not much is known about these kinds of interactions. To build an understanding of the role of non-Watson-Crick pairing on DNA behavior near surfaces, one requires basic information on DNA base pair stacking and hydrogen-bonding interactions. All-atom molecular simulations of DNA bases in two cases--in bulk water and strongly adsorbed on a graphite surface--are conducted to study the relative strengths of stacking and hydrogen bond interactions for each of the 10 possible combinations of base pairs. The key information obtained from these simulations is the free energy as a function of distance between two bases in a pair. We find that stacking interactions exert the dominant influence on the stability of DNA base pairs in bulk water as expected. The strength of stability for these stacking interactions is found to decrease in the order Gua-Gua > Ade-Gua > Ade-Ade > Gua-Thy > Gua-Cyt > Ade-Thy > Ade-Cyt > Thy-Thy > Cyt-Thy > Cyt-Cyt. On the other hand, mutual interactions of surface-adsorbed base pairs are stabilized mostly by hydrogen-bonding interactions in the order Gua-Cyt > Ade-Gua > Ade-Thy > Ade-Ade > Cyt-Thy > Gua-Gua > Cyt-Cyt > Ade-Cyt > Thy-Thy > Gua-Thy. Interestingly, several non-Watson-Crick base pairings, which are commonly ignored, have similar stabilization free energies due to interbase hydrogen bonding as Watson-Crick pairs. This clearly highlights the importance of non-Watson-Crick base pairing in the development of secondary structures of oligonucleotides near surfaces.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Paul, Debamita; Chatterjee, Abhishek; Begley, Tadhg P.
2010-11-15
THI6 is a bifunctional enzyme found in the thiamin biosynthetic pathway in eukaryotes. The N-terminal domain of THI6 catalyzes the ligation of the thiamin thiazole and pyrimidine moieties to form thiamin phosphate, and the C-terminal domain catalyzes the phosphorylation of 4-methyl-5-hydroxyethylthiazole in a salvage pathway. In prokaryotes, thiamin phosphate synthase and 4-methyl-5-hydroxyethylthiazole kinase are separate gene products. Here we report the first crystal structure of a eukaryotic THI6 along with several complexes that characterize the active sites responsible for the two chemical reactions. THI6 from Candida glabrata is a homohexamer in which the six protomers form a cage-like structure. Eachmore » protomer is composed of two domains, which are structurally homologous to their monofunctional bacterial counterparts. Two loop regions not found in the bacterial enzymes provide interactions between the two domains. The structures of different protein-ligand complexes define the thiazole and ATP binding sites of the 4-methyl-5-hydroxyethylthiazole kinase domain and the thiazole phosphate and 4-amino-5-hydroxymethyl-2-methylpyrimidine pyrophosphate binding sites of the thiamin phosphate synthase domain. Our structural studies reveal that the active sites of the two domains are 40 {angstrom} apart and are not connected by an obvious channel. Biochemical studies show 4-methyl-5-hydroxyethylthiazole phosphate is a substrate for THI6; however, adenosine diphospho-5{beta}-ethyl-4-methylthiazole-2-carboxylic acid, the product of THI4, is not a substrate for THI6. This suggests that an unidentified enzyme is necessary to produce the substrate for THI6 from the THI4 product.« less
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Mangia, Silvia; Giove, Federico; Tkáč, Ivan; Logothetis, Nikos K.; Henry, Pierre-Gilles; Olman, Cheryl A.; Maraviglia, Bruno; Di Salle, Francesco; Uğurbil, Kâmil
2009-01-01
Unraveling the energy metabolism and the hemodynamic outcomes of excitatory and inhibitory neuronal activity is critical not only for our basic understanding of overall brain function, but also for the understanding of many brain disorders. Methodologies of magnetic resonance spectroscopy (MRS) and magnetic resonance imaging (MRI) are powerful tools for the non-invasive investigation of brain metabolism and physiology. However, the temporal and spatial resolution of in vivo MRS and MRI is not suitable to provide direct evidence for hypotheses that involve metabolic compartmentalization between different cell types, or to untangle the complex neuronal micro-circuitry which results in changes of electrical activity. This review aims at describing how the current models of brain metabolism, mainly built on the basis of in vitro evidence, relate to experimental findings recently obtained in vivo by 1H MRS, 13C MRS and MRI. The hypotheses related to the role of different metabolic substrates, the metabolic neuron-glia interactions, along with the available theoretical predictions of the energy budget of neurotransmission, will be discussed. In addition, the cellular and network mechanisms that characterize different types of increased and suppressed neuronal activity will be considered within the sensitivity-constraints of MRS and MRI. PMID:19002199
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Antibacterial and efflux pump inhibitors of thymol and carvacrol against food-borne pathogens.
Miladi, Hanene; Zmantar, Tarek; Chaabouni, Yassine; Fedhila, Kais; Bakhrouf, Amina; Mahdouani, Kacem; Chaieb, Kamel
2016-10-01
In this study thymol (THY) and carvacrol (CAR), two monoterpenic phenol produced by various aromatic plants, was tested for their antibacterial and efflux pump inhibitors potencies against a panel of clinical and foodborne pathogenes. Our results demonstrated a substantial susceptibility of the tested bacteria toward THY and CAR. Especially, THY displayed a strong inhibitory activity (MIC's values ranged from 32 to 64 μg/mL) against the majority of the tested strains compared to CAR. Moreover, a significant reduction in MIC's of TET and benzalkonium chloride (QAC) were noticed when tested in combinations with THY and CAR. Their synergic effect was more significant in the case of THY which resulted a reduction of MIC's values of TET (2-8 fold) and QAC (2-8 fold). We noted also that THY and CAR inhibited the ethidium bromide (EtBr) cell efflux in a concentration-dependent manner. The rate of EtBr accumulation in food-borne pathogen was enhanced with THY and CAR (0, 250 and 500 μg/mL). The lowest concentration causing 50% of EtBr efflux inhibition (IC 50) was noticed in Salmonella enteritidis (1129) at 150 μg/mL of THY and 190 μg/mL of CAR respectively. These findings indicate that THY and CAR may serve as potential sources of efflux pump inhibitor in food-borne pathogens. Copyright © 2016 Elsevier Ltd. All rights reserved.
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A confirmatory factor analytic validation of the Tinnitus Handicap Inventory.
Kleinstäuber, Maria; Frank, Ina; Weise, Cornelia
2015-03-01
Because the postulated three-factor structure of the internationally widely used Tinnitus Handicap Inventory (THI) has not been confirmed yet by a confirmatory factor analytic approach this was the central aim of the current study. From a clinical setting, N=373 patients with chronic tinnitus completed the THI and further questionnaires assessing tinnitus-related and psychological variables. In order to analyze the psychometric properties of the THI, confirmatory factor analysis (CFA) and correlational analyses were conducted. CFA provided a statistically significant support for a better fit of the data to the hypothesized three-factor structure (RMSEA=.049, WRMR=1.062, CFI=.965, TLI=.961) than to a general factor model (RMSEA=.062, WRMR=1.258, CFI=.942, TLI=.937). The calculation of Cronbach's alpha as indicator of internal consistency revealed satisfactory values (.80-.91) with the exception of the catastrophic subscale (.65). High positive correlations of the THI and its subscales with other measures of tinnitus distress, anxiety, and depression, high negative correlations with tinnitus acceptance, moderate positive correlations with anxiety sensitivity, sleeping difficulties, tinnitus loudness, and small correlations with the Big Five personality dimensions confirmed construct validity. Results show that the THI is a highly reliable and valid measure of tinnitus-related handicap. In contrast to results of previous exploratory analyses the current findings speak for a three-factor in contrast to a unifactorial structure. Future research is needed to replicate this result in different tinnitus populations. Copyright © 2015 Elsevier Inc. All rights reserved.
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Thymosin From Bombyx mori Is Down-Regulated in Expression by BmNPV Exhibiting Antiviral Activity.
Zhang, Chen; Wang, Yongdi; Fang, Qiang; Xu, Minlin; Lv, Mengyuan; Liao, Jinxu; Li, Si; Nie, Zuoming; Zhang, Wenping
2016-01-01
Thymosins have been highly conserved during evolution. These hormones exist in many animal species and play an essential role in many biological events. However, little is known regarding the physiological function of silkworm Bombyx mori thymosin (BmTHY). In this study, we investigated the expression pattern of BmTHY in a Bombyx mori larval ovarian cell line (BmN) challenged with Bombyx mori nuclear polyhydrosis virus (BmNPV) and the antiviral effect of recombinant BmTHY (rBmTHY) for Bombyx mori against BmNPV. Western-blot assay and qRT-PCR analysis revealed that the level of BmTHY protein expression and transcription decreased over time when BmN cells were infected by BmNPV. Treatment with endotoxin-free rBmTHY led to a significant reduction in viral titer in the supernatant of BmN cells challenged with BmNPV. The results from antiviral tests performed in vitro and in vivo showed that endotoxin-free rBmTHY improved the survival rate of Bombyx mori infected with BmNPV. These findings suggest that BmTHY exerts immunomodulatory effects on Bombyx mori, rendering them resistant to viral infection. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America.
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Heat stress effects on Holstein dairy cows' rumination.
Moretti, R; Biffani, S; Chessa, S; Bozzi, R
2017-12-01
The objective of this study was to investigate the relationship between temperature-humidity index (THI) and rumination time (RT) in order to possibly exploit it as a useful tool for animal welfare improvement. During summer 2015 (1 June to 31 August), data from an Italian Holstein dairy farm located in the North of Italy were collected along with environmental data (i.e. ambient temperature and relative humidity) recorded with a weather station installed inside the barn. Rumination data were collected through the Heatime® HR system (SCR Engineers Ltd., Hadarim, Netanya, Israel), an automatic system composed of a neck collar with a Tag that records the RT and activity of each cow. A significant negative correlation was observed between RT and THI. Mixed linear models were fitted, including animal and test day as random effects, and parity, milk production level and date of last calving as fixed effects. A statistically significant effect of THI on RT was identified, with RT decreasing as THI increased.
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Norman, J C; McGee, M G; Fuqua, J M; Igo, S R; Turner, S A; Sterling, R; Urrutia, C O; Frazier, O H; Clay, W C; Chambers, J A
1983-02-01
A long-term, implantable, electrically actuated left ventricular assist system (THI/Gould LVAS) is being developed and characterized in vitro and in vivo for utilization in patients with end-stage heart disease. This system consists of five major components: a long-term, implantable blood pump (THI E-type ALVAD); an electrical-mechanical energy converter (Gould Model V); a control unit with batteries; a volume compensation system; and an external power supply and monitoring unit. Two of these components (blood pump and electrical-mechanical energy converter) have been integrated, and are undergoing chronic in vivo evaluations in calves. Thus far, 44 pneumatically and electrically actuated THI/Gould LVAS evaluations have been performed. This experience has resulted in greater than 6.5 years of actuation in vivo, with durations exceeding 1 year. System in vivo performance in terms of durability, mechanical reliability, hemodynamic effectiveness, and biocompatibility has been satisfactory. Demonstration of long-term (2-year) effectiveness in supporting the circulation is the ultimate goal.
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Nilsen, Linn H; Melø, Torun M; Saether, Oddbjørn; Witter, Menno P; Sonnewald, Ursula
2012-11-01
We investigated metabolite levels during the progression of pathology in McGill-R-Thy1-APP rats, a transgenic animal model of Alzheimer's disease, and in healthy age-matched controls. Rats were subjected to in vivo (1) H magnetic resonance spectroscopy (MRS) of the dorsal hippocampus at age 3, 9 and 12 months and of frontal cortex at 9 and 12 months. At 3 months, a stage in which only Aβ oligomers are present, lower glutamate, myo-inositol and total choline content were apparent in McGill-R-Thy1-APP rats. At age 9 months, lower levels of glutamate, GABA, N-acetylaspartate and total choline and elevated myo-inositol and taurine were found in dorsal hippocampus, whereas lower levels of glutamate, GABA, glutamine and N-acetylaspartate were found in frontal cortex. At age 12 months, only the taurine level was significantly different in dorsal hippocampus, whereas taurine, myo-inositol, N-acetylaspartate and total creatine levels were significantly higher in frontal cortex. McGill-R-Thy1-APP rats did not show the same changes in metabolite levels with age as displayed in the controls, and overall, prominent and complex metabolite differences were evident in this transgenic rat model of Alzheimer's disease. The findings also demonstrate that in vivo (1) H MRS is a powerful tool to investigate disease-related metabolite changes in the brain. © 2012 The Authors Journal of Neurochemistry © 2012 International Society for Neurochemistry.
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Zhao, Xiao-Su; Fu, Wing-Yu; Chien, Winnie W. Y.; Li, Zhen; Fu, Amy K. Y.; Ip, Nancy Y.
2014-01-01
Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase, which plays critical roles in a wide spectrum of neuronal functions including neuronal survival, neurite outgrowth, and synapse development and plasticity. Cdk5 activity is controlled by its specific activators: p35 or p39. While knockout studies reveal that Cdk5/p35 is critical for neuronal migration during early brain development, functions of Cdk5/p35 have been unraveled through the identification of the interacting proteins of p35, most of which are Cdk5/p35 substrates. However, it remains unclear whether p35 can regulate neuronal functions independent of Cdk5 activity. Here, we report that a nuclear protein, nuclear hormone receptor coregulator (NRC)-interacting factor 1 (NIF-1), is a new interacting partner of p35. Interestingly, p35 regulates the functions of NIF-1 independent of Cdk5 activity. NIF-1 was initially discovered as a transcriptional regulator that enhances the transcriptional activity of nuclear hormone receptors. Our results show that p35 interacts with NIF-1 and regulates its nucleocytoplasmic trafficking via the nuclear export pathway. Furthermore, we identified a nuclear export signal on p35; mutation of this site or blockade of the CRM1/exportin-dependent nuclear export pathway resulted in the nuclear accumulation of p35. Intriguingly, blocking the nuclear export of p35 attenuated the nuclear accumulation of NIF-1. These findings reveal a new p35-dependent mechanism in transcriptional regulation that involves the nucleocytoplasmic shuttling of transcription regulators. PMID:25329792
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Zhao, Xiao-Su; Fu, Wing-Yu; Chien, Winnie W Y; Li, Zhen; Fu, Amy K Y; Ip, Nancy Y
2014-01-01
Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase, which plays critical roles in a wide spectrum of neuronal functions including neuronal survival, neurite outgrowth, and synapse development and plasticity. Cdk5 activity is controlled by its specific activators: p35 or p39. While knockout studies reveal that Cdk5/p35 is critical for neuronal migration during early brain development, functions of Cdk5/p35 have been unraveled through the identification of the interacting proteins of p35, most of which are Cdk5/p35 substrates. However, it remains unclear whether p35 can regulate neuronal functions independent of Cdk5 activity. Here, we report that a nuclear protein, nuclear hormone receptor coregulator (NRC)-interacting factor 1 (NIF-1), is a new interacting partner of p35. Interestingly, p35 regulates the functions of NIF-1 independent of Cdk5 activity. NIF-1 was initially discovered as a transcriptional regulator that enhances the transcriptional activity of nuclear hormone receptors. Our results show that p35 interacts with NIF-1 and regulates its nucleocytoplasmic trafficking via the nuclear export pathway. Furthermore, we identified a nuclear export signal on p35; mutation of this site or blockade of the CRM1/exportin-dependent nuclear export pathway resulted in the nuclear accumulation of p35. Intriguingly, blocking the nuclear export of p35 attenuated the nuclear accumulation of NIF-1. These findings reveal a new p35-dependent mechanism in transcriptional regulation that involves the nucleocytoplasmic shuttling of transcription regulators.
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Genetics of heat tolerance for milk yield and quality in Holsteins.
Santana, M L; Bignardi, A B; Pereira, R J; Stefani, G; El Faro, L
2017-01-01
Tropical and sub-tropical climates are characterized by high temperature and humidity, during at least part of the year. Consequently, heat stress is common in Holstein cattle and productive and reproductive losses are frequent. Our objectives were as follows: (1) to quantify losses in production and quality of milk due to heat stress; (2) to estimate genetic correlations within and between milk yield (MY) and milk quality traits; and (3) to evaluate the trends of genetic components of tolerance to heat stress in multiple lactations of Brazilian Holstein cows. Thus, nine analyses using two-trait random regression animal models were carried out to estimate variance components and genetic parameters over temperature-humidity index (THI) values for MY and milk quality traits (three lactations: MY×fat percentage (F%), MY×protein percentage (P%) and MY×somatic cell score (SCS)) of Brazilian Holstein cattle. It was demonstrated that the effects of heat stress can be harmful for traits related to milk production and milk quality of Holstein cattle even though most herds were maintained in a modified environment, for example, with fans and sprinklers. For MY, the effect of heat stress was more detrimental in advanced lactations (-0.22 to -0.52 kg/day per increase of 1 THI unit). In general, the mean heritability estimates were higher for lower THI values and longer days in milk for all traits. In contrast, the heritability estimates for SCS increased with increasing THI values in the second and third lactation. For each trait studied, lower genetic correlations (different from unity) were observed between opposite extremes of THI (THI 47 v. THI 80) and in advanced lactations. The genetic correlations between MY and milk quality trait varied across the THI scale and lactations. The genotype×environment interaction due to heat stress was more important for MY and SCS, particularly in advanced lactations, and can affect the genetic relationship between MY and milk quality traits. Selection for higher MY, F% or P% may result in a poor response of the animals to heat stress, as a genetic antagonism was observed between the general production level and specific ability to respond to heat stress for these traits. Genetic trends confirm the adverse responses in the genetic components of heat stress over the years for milk production and quality. Consequently, the selection of Holstein cattle raised in modified environments in both tropical and sub-tropical regions should take into consideration the genetic variation in heat stress.
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NASA Astrophysics Data System (ADS)
Ahirwar, Maneesh Kumar; Kataktalware, Mukund Amritrao; Ramesha, Kerekoppa Puttaiah; Pushpadass, Heartwin Amaladhas; Jeyakumar, Sakthivel; Revanasiddu, Deginal; Kour, Reen Jagish; Nath, Sapna; Nagaleekar, Anand Kumar; Nazar, Sayyad
2017-12-01
The aim of the present study was to examine the effects of non-genetic factors on scrotal thermographic profile viz., proximal pole temperature (PPT °C), mid pole temperature (MPT °C), distal pole temperature (DPT °C) and ocular temperature (OcT) of Murrah ( Bubalus bubalis) breeding bulls. A total of 109 buffalo bulls, maintained at three semen stations (SS), were monitored for scrotal surface and ocular temperatures using infrared thermography twice daily during rainy, winter and summer seasons using an FLIR i5 infrared camera and temperatures were measured. Thermograms were analysed by FLIR QuickReport v.1.2 SP2 software. Statistical analysis revealed that semen station, season, temperature humidity index (THI), housing system and timing of observations had significant ( P < 0.05) effect on scrotal surface temperature (SST) and OcT. In SS-I, the PPT and MPT were significantly ( P < 0.05) higher as compared to SS-II and SS-III. THI had significant ( P < 0.05) effect on SST and OcT, whereas PPT (°C), MPT (°C), DPT (°C) and OcT (°C) values during high THI (>80.88; <0.05) period were higher as compared to medium THI period (70.06-80.88) and during low THI period (<70.06). Temperature gradient (TG) of the testes was significantly ( P < 0.05) higher during low THI period (4.50 ± 0.06 °C) as compared to medium THI (2.38 ± 0.03 °C) and high THI (1.61 ± 0.05 °C). Season of the year had a significant effect ( P < 0.05) on the SST and OcT. During the rainy season, PPT (34.50 ± 0.09 °C), MPT (33.44 ± 0.12 °C) and DPT (32.11 ± 0.15 °C) were significantly ( P < 0.05) higher as compared to winter and summer seasons. Age of the bulls had non-significant effect on SST and OcT but had a marked influence on thermal profile of scrotum. It could be concluded semen station, season, temperature humidity index, housing system and timing of observations had a significant influence on scrotal surface temperature. The monitoring of scrotal surface temperature by infrared thermography was found to be useful in evaluating the effects of thermal stress on physiology and health of buffalo bulls.
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Ahirwar, Maneesh Kumar; Kataktalware, Mukund Amritrao; Ramesha, Kerekoppa Puttaiah; Pushpadass, Heartwin Amaladhas; Jeyakumar, Sakthivel; Revanasiddu, Deginal; Kour, Reen Jagish; Nath, Sapna; Nagaleekar, Anand Kumar; Nazar, Sayyad
2017-12-01
The aim of the present study was to examine the effects of non-genetic factors on scrotal thermographic profile viz., proximal pole temperature (PPT °C), mid pole temperature (MPT °C), distal pole temperature (DPT °C) and ocular temperature (OcT) of Murrah (Bubalus bubalis) breeding bulls. A total of 109 buffalo bulls, maintained at three semen stations (SS), were monitored for scrotal surface and ocular temperatures using infrared thermography twice daily during rainy, winter and summer seasons using an FLIR i5 infrared camera and temperatures were measured. Thermograms were analysed by FLIR QuickReport v.1.2 SP2 software. Statistical analysis revealed that semen station, season, temperature humidity index (THI), housing system and timing of observations had significant (P < 0.05) effect on scrotal surface temperature (SST) and OcT. In SS-I, the PPT and MPT were significantly (P < 0.05) higher as compared to SS-II and SS-III. THI had significant (P < 0.05) effect on SST and OcT, whereas PPT (°C), MPT (°C), DPT (°C) and OcT (°C) values during high THI (>80.88; <0.05) period were higher as compared to medium THI period (70.06-80.88) and during low THI period (<70.06). Temperature gradient (TG) of the testes was significantly (P < 0.05) higher during low THI period (4.50 ± 0.06 °C) as compared to medium THI (2.38 ± 0.03 °C) and high THI (1.61 ± 0.05 °C). Season of the year had a significant effect (P < 0.05) on the SST and OcT. During the rainy season, PPT (34.50 ± 0.09 °C), MPT (33.44 ± 0.12 °C) and DPT (32.11 ± 0.15 °C) were significantly (P < 0.05) higher as compared to winter and summer seasons. Age of the bulls had non-significant effect on SST and OcT but had a marked influence on thermal profile of scrotum. It could be concluded semen station, season, temperature humidity index, housing system and timing of observations had a significant influence on scrotal surface temperature. The monitoring of scrotal surface temperature by infrared thermography was found to be useful in evaluating the effects of thermal stress on physiology and health of buffalo bulls.
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Digesting dietary miRNA therapeutics
USDA-ARS?s Scientific Manuscript database
Hippocrates famously advised, "Let food be thy medicine and thy medicine be thy food." Numerous plant-derived compounds are used as cancer therapeutics including antimitotics, topoisomerase inhibitors, and kinase inhibitors. Here we will review emerging evidence suggesting that diet derived small RN...
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Likavec, Tasha; Pires, Alda F.A.; Funk, Julie A.
2016-01-01
The objective of this study was to describe the association between thermal measures in the barn environment (pen temperature and humidity) and fecal shedding of Salmonella in dairy cattle. A repeated cross-sectional study was conducted within a commercial dairy herd located in the midwestern United States. Five pooled fecal samples were collected monthly from each pen for 9 mo and submitted for microbiological culture. Negative binomial regression methods were used to test the association [incidence rate ratio (IRR)] between Salmonella pen status (the count of Salmonella-positive pools) and thermal environmental parameters [average temperature and temperature humidity index (THI)] for 3 time periods (48 h, 72 h, and 1 wk) before fecal sampling. Salmonella was cultured from 10.8% [39/360; 95% confidence interval (CI): 7.8% to 14.5%] of pooled samples. The highest proportion of positive pools occurred in August. The IRR ranged from 1.26 (95% CI: 1.15 to 1.39, THI 1 wk) to 4.5 (95% CI: 2.13 to 9.51, heat exposure 1 wk) across all thermal parameters and lag time periods measured. For example, the incidence rate of Salmonella-positive pools increased by 54% for every 5°C increment in average temperature (IRR = 1.54; 95% CI: 1.29 to 1.85) and 29% for every 5-unit increase in THI (IRR = 1.29; 95% CI: 1.16 to 1.42) during the 72 h before sampling. The incidence rate ratio for pens exposed to higher temperatures (> 25°C) was 4.5 times (95% CI: 2.13 to 9.51) the incidence rate ratio for pens exposed to temperatures < 25°C in the 72 h before sampling. Likewise, the incidence rate ratio for pens exposed to THI > 70 was 4.23 times greater (95% CI: 2.1 to 8.28) than when the THI was < 70 in the 72 h before sampling. An association was found between the thermal environment and Salmonella shedding in dairy cattle. Further research is warranted in order to fully understand the component risks associated with the summer season and increased Salmonella shedding. PMID:27408330
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NASA Astrophysics Data System (ADS)
El-Tarabany, Mahmoud Salah
2016-07-01
The aim of this study was to investigate the effect of temperature-humidity index (THI) level on productive parameters, welfare, and immunity in Japanese quails. One hundred and eighty (180) birds of Japanese quail, 14 weeks old, were used. Birds were divided randomly into three equal groups, control (at low THI, less than 70), H1 (at moderate THI, 70-75), and H2 (at high THI, 76-80). Birds in the control group had higher body weight (281.2 g, p = 0.001), egg mass (745 g, p = 0.001), fertility (85.4 %, p = 0.039), hatchability (80.4 %, p = 0.001), and immune response titer to Newcastle disease virus ( p = 0.031), compared with H2 group. Furthermore, the thermoneutral group had higher internal egg quality score [albumen height (5.14 mm, p = 0.001), yolk height (10.88 mm, p = 0.015), yolk index (42.32 %, p = 0.039), and Haugh unit (92.67, p = 0.001)]. Nevertheless, there were no significant differences in fertility percentage, immune response, and corticosterone concentration between control and H1 group. Birds in the H2 group had the lowest total leucocytic count and lymphocyte percentage ( p = 0.001 and 0.020, respectively) but the highest H/L ratio (0.83, p = 0.001). Corticosterone concentration was lower in control and H1 groups (5.49 and 6.41 ng/mL, respectively, p = 0.024) than that in H2 group. Japanese quail exposed to heat stress revealed drop in production and immunological parameters, as well as a detrimental effects on welfare. Thus, practical approaches might be used to reduce the detrimental effects of greater THI level.
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Yan, Y; Zhao, J; Cao, C; Jia, Z; Zhou, N; Han, S; Wang, Y; Xu, Y; Zhao, J; Yan, Y; Cui, H
2014-10-10
Tetramethylpyrazine (TMP) is an active compound extracted from the traditional Chinese medicinal herb Chuanxiong. Recently, it has been reported that TMP enhances neurogenesis, and promotes neural stem cell differentiation toward neurons. However, its molecular basis remains unknown. Topoisomerase IIβ (TopoIIβ) is a nuclear enzyme with an essential role in neuronal development. This study aimed to investigate whether TopoIIβ is involved in TMP-induced neuronal differentiation. We examined the effect of TMP on neuronal differentiation of SH-SY5Y cells. It was found that TMP inhibited cell proliferation and induced G0/G1 cell cycle arrest. TMP promoted SH-SY5Y cells to differentiate toward post-mitotic neurons characterized by long, out-branched neurites and up-regulated neuronal markers, microtubule-associated protein 2 (MAP2) and tau. Meanwhile, we demonstrated that TopoIIβ was highly expressed following TMP treatment. To unravel how TMP affects TopoIIβ expression, two chromatin active markers, acetylated histone H3 (Ac-H3) and acetylated histone H4 (Ac-H4) were examined in this study. Our data showed that the levels of Ac-H3 and Ac-H4 were positively correlated with TopoIIβ expression in the processes of neuronal differentiation. We further performed chromatin immunoprecipitation (ChIP) analysis and identified that TMP enhanced the recruitment of Ac-H3 and Ac-H4 to the TopoIIβ gene promoter region. Therefore, we concluded that TMP may stimulate neuronal differentiation of SH-SY5Y cells through epigenetic regulation of TopoIIβ. These results suggest a novel molecular mechanism underlying TMP-promoted neuronal differentiation. Copyright © 2014 IBRO. Published by Elsevier Ltd. All rights reserved.
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Görtelmeyer, Roman; Schmidt, Jürgen; Suckfüll, Markus; Jastreboff, Pawel; Gebauer, Alexander; Krüger, Hagen; Wittmann, Werner
2011-08-01
To evaluate the reliability, dimensionality, predictive validity, construct validity, and sensitivity to change of the THI-12 total and sub-scales as diagnostic aids to describe and quantify tinnitus-evoked reactions and evaluate treatment efficacy. Explorative analysis of the German tinnitus handicap inventory (THI-12) to assess potential sensitivity to tinnitus therapy in placebo-controlled randomized studies. Correlation analysis, including Cronbach's coefficient α and explorative common factor analysis (EFA), was conducted within and between assessments to demonstrate the construct validity, dimensionality, and factorial structure of the THI-12. N = 618 patients suffering from subjective tinnitus who were to be screened to participate in a randomized, placebo-controlled, 16-week, longitudinal study. The THI-12 can reliably diagnose tinnitus-related impairments and disabilities and assess changes over time. The test-retest coefficient for neighboured visits was r > 0.69, the internal consistency of the THI-12 total score was α ≤ 0.79 and α ≤ 0.89 at subsequent visits. Predictability of THI-12 total score and overall variance increased with successive measurements. The three-factorial structure allowed for evaluation of factors that affect aspects of patients' health-related quality of life. The THI-12, with its three-factorial structure, is a simple, reliable, and valid instrument for the diagnosis and assessment of tinnitus and associated impairment over time.
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Cellular Insulin Resistance Disrupts Leptin-Mediated Control of Neuronal Signaling and Transcription
Nazarians-Armavil, Anaies; Menchella, Jonathan A.
2013-01-01
Central resistance to the actions of insulin and leptin is associated with the onset of obesity and type 2 diabetes mellitus, whereas leptin and insulin signaling is essential for both glucose and energy homeostasis. Although it is known that leptin resistance can lead to attenuated insulin signaling, whether insulin resistance can lead to or exacerbate leptin resistance is unknown. To investigate the molecular events underlying crosstalk between these signaling pathways, immortalized hypothalamic neuronal models, rHypoE-19 and mHypoA-2/10, were used. Prolonged insulin exposure was used to induce cellular insulin resistance, and thereafter leptin-mediated regulation of signal transduction and gene expression was assessed. Leptin directly repressed agouti-related peptide mRNA levels but induced urocortin-2, insulin receptor substrate (IRS)-1, IRS2, and IR transcription, through leptin-mediated phosphatidylinositol 3-kinase/Akt activation. Neuronal insulin resistance, as assessed by attenuated Akt phosphorylation, blocked leptin-mediated signal transduction and agouti-related peptide, urocortin-2, IRS1, IRS2, and insulin receptor synthesis. Insulin resistance caused a substantial decrease in insulin receptor protein levels, forkhead box protein 1 phosphorylation, and an increase in suppressor of cytokine signaling 3 protein levels. Cellular insulin resistance may cause or exacerbate neuronal leptin resistance and, by extension, obesity. It is essential to unravel the effects of neuronal insulin resistance given that both peripheral, as well as the less widely studied central insulin resistance, may contribute to the development of metabolic, reproductive, and cardiovascular disorders. This study provides improved understanding of the complex cellular crosstalk between insulin-leptin signal transduction that is disrupted during neuronal insulin resistance. PMID:23579487
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2014-01-01
Background In dementia with Lewy bodies (DLB) abnormal interactions between α-synuclein (α-syn) and beta amyloid (Aβ) result in selective degeneration of neurons in the neocortex, limbic system and striatum. However, factors rendering these neurons selectively vulnerable have not been fully investigated. The metabotropic glutamate receptor 5 (mGluR5) has been shown to be up regulated in DLB and might play a role as a mediator of the neurotoxic effects of Aβ and α-syn in vulnerable neuronal populations. In this context, the main objective of the present study was to investigate the role of mGluR5 as a mediator of the neurotoxic effects of α-syn and Aβ in the hippocampus. Results We generated double transgenic mice over-expressing amyloid precursor protein (APP) and α-syn under the mThy1 cassette and investigated the relationship between α-syn cleavage, Aβ, mGluR5 and neurodegeneration in the hippocampus. We found that compared to the single tg mice, the α-syn/APP tg mice displayed greater accumulation of α-syn and mGluR5 in the CA3 region of the hippocampus compared to the CA1 and other regions. This was accompanied by loss of CA3 (but not CA1) neurons in the single and α-syn/APP tg mice and greater loss of MAP 2 and synaptophysin in the CA3 in the α-syn/APP tg. mGluR5 gene transfer using a lentiviral vector into the hippocampus CA1 region resulted in greater α-syn accumulation and neurodegeneration in the single and α-syn/APP tg mice. In contrast, silencing mGluR5 with a lenti-shRNA protected neurons in the CA3 region of tg mice. In vitro, greater toxicity was observed in primary hippocampal neuronal cultures treated with Aβ oligomers and over-expressing α-syn; this effect was attenuated by down-regulating mGluR5 with an shRNA lentiviral vector. In α-syn-expressing neuronal cells lines, Aβ oligomers promoted increased intracellular calcium levels, calpain activation and α-syn cleavage resulting in caspase-3-dependent cell death. Treatment with pharmacological mGluR5 inhibitors such as 2-Methyl-6-(phenylethynyl)pyridine (MPEP) and 3-((2-Methyl-4-thiazolyl)ethynyl)pyridine (MTEP) attenuated the toxic effects of Aβ in α-syn-expressing neuronal cells. Conclusions Together, these results support the possibility that vulnerability of hippocampal neurons to α-syn and Aβ might be mediated via mGluR5. Moreover, therapeutical interventions targeting mGluR5 might have a role in DLB. PMID:24885390
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Yi, Chun-Xia; Sun, Ning; Ackermans, Mariette T; Alkemade, Anneke; Foppen, Ewout; Shi, Jing; Serlie, Mireille J; Buijs, Ruud M; Fliers, Eric; Kalsbeek, Andries
2010-07-01
The unraveling of the elaborate brain networks that control glucose metabolism presents one of the current challenges in diabetes research. Within the central nervous system, the hypothalamus is regarded as the key brain area to regulate energy homeostasis. The aim of the present study was to investigate the hypothalamic mechanism involved in the hyperglycemic effects of the neuropeptide pituitary adenylyl cyclase-activating polypeptide (PACAP). Endogenous glucose production (EGP) was determined during intracerebroventricular infusions of PACAP-38, vasoactive intestinal peptide (VIP), or their receptor agonists. The specificity of their receptors was examined by coinfusions of receptor antagonists. The possible neuronal pathway involved was investigated by 1) local injections in hypothalamic nuclei, 2) retrograde neuronal tracing from the thoracic spinal cord to hypothalamic preautonomic neurons together with Fos immunoreactivity, and 3) specific hepatic sympathetic or parasympathetic denervation to block the autonomic neuronal input to liver. Intracerebroventricular infusion of PACAP-38 increased EGP to a similar extent as a VIP/PACAP-2 (VPAC2) receptor agonist, and intracerebroventricular administration of VIP had significantly less influence on EGP. The PACAP-38 induced increase of EGP was significantly suppressed by preinfusion of a VPAC2 but not a PAC1 receptor antagonist, as well as by hepatic sympathetic but not parasympathetic denervation. In the hypothalamus, Fos immunoreactivity induced by PACAP-38 was colocalized within autonomic neurons in paraventricular nuclei projecting to preganglionic sympathetic neurons in the spinal cord. Local infusion of PACAP-38 directly into the PVN induced a significant increase of EGP. This study demonstrates that PACAP-38 signaling via sympathetic preautonomic neurons located in the paraventricular nucleus is an important component in the hypothalamic control of hepatic glucose production.
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Dash, Soumya; Chakravarty, A K; Singh, Avtar; Shivahre, Pushp Raj; Upadhyay, Arpan; Sah, Vaishali; Singh, K Mahesh
2015-03-01
The aim of the present study was to assess the influence of temperature and humidity prevalent under subtropical climate on the breeding values for fertility traits viz. service period (SP), pregnancy rate (PR) and conception rate (CR) of Murrah buffaloes in National Dairy Research Institute (NDRI) herd. Fertility data on 1379 records of 581 Murrah buffaloes spread over four lactations and climatic parameters viz. dry bulb temperature and relative humidity (RH) spanned over 20 years (1993-2012) were collected from NDRI and Central Soil and Salinity Research Institute, Karnal, India. Monthly average temperature humidity index (THI) values were estimated. Threshold THI value affecting fertility traits was identified by fixed least-squares model analysis. Three zones of non-heat stress, heat stress and critical heat stress zones were developed in a year. The genetic parameters heritability (h(2)) and repeatability (r) of each fertility trait were estimated. Genetic evaluation of Murrah buffaloes was performed in each zone with respect to their expected breeding values (EBV) for fertility traits. Effect of THI was found significant (p<0.001) on all fertility traits with threshold THI value identified as 75. Based on THI values, a year was classified into three zones: Non heat stress zone(THI 56.71-73.21), HSZ (THI 75.39-81.60) and critical HSZ (THI 80.27-81.60). The EBVfor SP, PR, CR were estimated as 138.57 days, 0.362 and 69.02% in non-HSZ while in HSZ EBV were found as 139.62 days, 0.358 and 68.81%, respectively. EBV for SP was increased to 140.92 days and for PR and CR, it was declined to 0.357 and 68.71% in critical HSZ. The negative effect of THI was observed on EBV of fertility traits under the non-HSZ and critical HSZ Thus, the influence of THI should be adjusted before estimating the breeding values for fertility traits in Murrah buffaloes.
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Mollica, Richard F.; Chernoff, Miriam C.; Berthold, S. Megan; Lavelle, James; Lyoo, In Kyoon; Renshaw, Perry
2014-01-01
Little is known about the relationship between traumatic head injury (THI) and psychiatric morbidity in torture survivors. We examine the relationship between THI and depression, PTSD, post-concussive syndrome (PCS), disability and poor health status in Vietnamese ex-political detainees who survived incarceration in Vietnamese re-education camps. A community sample of ex-political detainees (n=337) and a non-THI, non-ex-detainee comparison group (n=82) were surveyed. 78% of the ex-political detainees had experienced THI. 90.6% of the ex-political detainees and 3.6% of the comparison group had experienced 7 or more trauma events. Depression and PTSD were greater in ex-detainees than the comparison group (40.9% vs 23.2% and 13.4% vs 0%). Dose-effect relationships for THI and trauma/torture in the ex-political detainee group were significant. Logistic regression in the pooled sample of ex-detainees and the comparison group confirmed the independent impact of THI from trauma/torture on psychiatric morbidity (OR for PTSD=22.4; 95% CI: 3.0-165.8). These results demonstrate important effects of THI on depression and PTSD in Vietnamese ex-detainees who have survived torture. PMID:24962448
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Patient-based outcomes in patients with primary tinnitus undergoing tinnitus retraining therapy.
Berry, Julie A; Gold, Susan L; Frederick, Ellen Alvarez; Gray, William C; Staecker, Hinrich
2002-10-01
To determine whether the Tinnitus Handicap Inventory (THI), a validated patient-based outcomes measure, may improve our ability to quantify impact and assess therapy for patients with tinnitus. Nonrandomized, prospective analysis of 32 patients undergoing tinnitus retraining therapy (TRT). Assessment tools included comprehensive audiology, a subjective self-assessment survey of tinnitus characteristics, and the THI. Tinnitus Handicap Inventory scores were assessed at baseline and 6 months following TRT. Baseline analysis revealed significant correlation between the subjective presence of hyperacusis and higher total, emotional, and catastrophic THI scores. Tinnitus Handicap Inventory scores correlated with subjective perception of overall tinnitus effect (P<.001). Mean pure-tone threshold average was 17.4 dB, and mean speech discrimination was 97.0%. There were no consistent correlations between baseline audiologic parameters and THI scores. Following 6 months of TRT, the total, emotional, functional, and catastrophic THI scores significantly improved (P<.001). Loudness discomfort levels also significantly improved (P< or =.02). There is significant improvement in self-perceived disability following TRT as measured by the THI. The results confirm the utility of the THI as a patient-based outcomes measure for quantifying treatment status in patients with primary tinnitus.
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Hypothalamic neurones governing glucose homeostasis.
Coppari, R
2015-06-01
The notion that the brain directly controls the level of glucose in the blood (glycaemia) independent of its known action on food intake and body weight has been known ever since 1849. That year, the French physiologist Dr Claude Bernard reported that physical puncture of the floor of the fourth cerebral ventricle rapidly leads to an increased level of sugar in the blood (and urine) in rabbits. Despite this important discovery, it took approximately 150 years before significant efforts aimed at understanding the underlying mechanism of brain-mediated control of glucose metabolism were made. Technological developments allowing for genetically-mediated manipulation of selected molecular pathways in a neurone-type-specific fashion unravelled the importance of specific molecules in specific neuronal populations. These neuronal pathways govern glucose metabolism in the presence and even in the absence of insulin. Also, a peculiarity of these pathways is that certain biochemically-defined neurones govern glucose metabolism in a tissue-specific fashion. © 2015 British Society for Neuroendocrinology.
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The Molecular Design of High-Performance Carbon Materials
2008-06-30
Thies MC. Control of mesophase pitch properties by supercritical fluid extraction. Carbon 1998; 36(7-8):953-61. 7] Zhuang M, Gast K, Thies MC...pitch with supercritical toluene. J Supercrit Fluids 1991; 4(1):7-14. 16] Herod AA, Bartle KD, Kandiyoti R. Characterization of heavy...MALDI, mass spectrometry, mesophase, extraction, supercritical , fractionation. 16. SECURITY CLASSIFICATION OF: a. REPORT u b. ABSTRACT u c. THIS
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Integration of autopatching with automated pipette and cell detection in vitro
Wu (吴秋雨), Qiuyu; Kolb, Ilya; Callahan, Brendan M.; Su, Zhaolun; Stoy, William; Kodandaramaiah, Suhasa B.; Neve, Rachael; Zeng, Hongkui; Boyden, Edward S.; Forest, Craig R.
2016-01-01
Patch clamp is the main technique for measuring electrical properties of individual cells. Since its discovery in 1976 by Neher and Sakmann, patch clamp has been instrumental in broadening our understanding of the fundamental properties of ion channels and synapses in neurons. The conventional patch-clamp method requires manual, precise positioning of a glass micropipette against the cell membrane of a visually identified target neuron. Subsequently, a tight “gigaseal” connection between the pipette and the cell membrane is established, and suction is applied to establish the whole cell patch configuration to perform electrophysiological recordings. This procedure is repeated manually for each individual cell, making it labor intensive and time consuming. In this article we describe the development of a new automatic patch-clamp system for brain slices, which integrates all steps of the patch-clamp process: image acquisition through a microscope, computer vision-based identification of a patch pipette and fluorescently labeled neurons, micromanipulator control, and automated patching. We validated our system in brain slices from wild-type and transgenic mice expressing channelrhodopsin 2 under the Thy1 promoter (line 18) or injected with a herpes simplex virus-expressing archaerhodopsin, ArchT. Our computer vision-based algorithm makes the fluorescent cell detection and targeting user independent. Compared with manual patching, our system is superior in both success rate and average trial duration. It provides more reliable trial-to-trial control of the patching process and improves reproducibility of experiments. PMID:27385800
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Translation, Adaptation and Cross Language Validation of Tinnitus Handicap Inventory in Urdu.
Aqeel, Muhammad; Ahmed, Ammar
2017-12-01
Tinnitus is characterized as a perception of numerous auditory sounds in absence of external stimulus. Tinnitus can have a considerable consequence on a person's quality of life, and is considered to be very complicated to quantify. The aim of this study was to investigate the reliability and validity of Urdu translation of the Tinnitus Handicap Inventory (THI) in Pakistan. It was designed to assess the presence of various auditory sounds without the external stimulus. Scale consisted of 25 items having three subscales functional, emotional, and catastrophic. The study comprised into two stages, preliminary and main studies. The results of preliminary study revealed that the overall scale had high internal consistency [alpha coefficient of Urdu version of THI (THI-U)= 0.99, alpha coefficient of English version of THI=0.98]. The overall scale had test-retest correlation over a fifteen days period of interval (0.99). Main study was performed on 110 tinnitus patients. The results of main study showed that the internal consistency and reliability of Urdu version was (α=0.93). The THI-U and its subscales demonstrated good internal consistency reliability ( α =0.81 to 0.86). High to moderate correlations were noted between tinnitus symptom ratings. A confirmatory factor analysis was used to validate the three subscales of THI-U, and high inter-correlations were found between the subscales also results revealed that a three-factor model for the THI-U was most tenable. The results displayed that the confirmatory factor analysis confirmed to validate the three subscales of THI-U. THI-U might present important information about precise facets of tinnitus distress along with diagnostic interviews in clinical practice.
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Krashes, Michael J; Shah, Bhavik P; Koda, Shuichi; Lowell, Bradford B
2013-10-01
Agouti-related peptide (AgRP) neurons of the hypothalamus release a fast transmitter (GABA) in addition to neuropeptides (neuropeptide Y [NPY] and Agouti-related peptide [AgRP]). This raises questions as to their respective functions. The acute activation of AgRP neurons robustly promotes food intake, while central injections of AgRP, NPY, or GABA agonist results in the marked escalation of food consumption with temporal variance. Given the orexigenic capability of all three of these neuroactive substances in conjunction with their coexpression in AgRP neurons, we looked to unravel their relative temporal role in driving food intake. After the acute stimulation of AgRP neurons with DREADD technology, we found that either GABA or NPY is required for the rapid stimulation of feeding, and the neuropeptide AgRP, through action on MC4 receptors, is sufficient to induce feeding over a delayed yet prolonged period. These studies help to elucidate the neurochemical mechanisms of AgRP neurons in controlling temporally distinct phases of eating. Copyright © 2013 Elsevier Inc. All rights reserved.
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Dynamics of the temperature-humidity index in the Mediterranean basin
NASA Astrophysics Data System (ADS)
Segnalini, Maria; Nardone, Alessandro; Bernabucci, Umberto; Vitali, Andrea; Ronchi, Bruno; Lacetera, Nicola
2011-03-01
The study was aimed at describing the temperature humidity index (THI) dynamics over the Mediterranean basin for the period 1951-2007. The THI combines temperature and humidity into a single value, and may help to predict the effects of environmental warmth in farm animals. In particular, on the basis of THI values, numerous studies have been performed to establish thresholds for heat stress in dairy cows. The THI was calculated by using monthly mean values of temperature and humidity obtained from the National Center for Environmental Prediction/National Center for Atmospheric Research reanalysis project. The analysis demonstrated a high degree of heterogeneity of THI patterns over the Mediterranean basin, a strong north-south gradient, and an overall warming during the study period, which was particularly marked during summer seasons. Results indicated that several areas of the basin present summer THI values which were unfavorable to cow welfare and productivity, and that risk of heat stress for cows is generally greater in the countries of the south coast of the basin. Furthermore, THI data from the summer 2003 revealed that severe positive anomalies may impact areas normally characterized by a favorable climate for animal production. In conclusion, THI dynamics should be taken into careful consideration by farmers and policy makers operating in Mediterranean countries when planning investments in the sector of animal production. The investments should at least partially be directed towards implementation of adaptation measures, which may help to alleviate the impact of hot on farm animals welfare, performance and health.
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Impaired reproduction in Japanese Black cattle under cold environmental conditions.
Nabenishi, H; Yamazaki, A
2017-06-01
Environmental factors such as the temperature-humidity index (THI) are known to affect reproductive parameters in cattle. Therefore, here, we examined whether there was any correlation between the THI and the reproductive performance of Japanese Black cattle by analysing the first-service conception rates of 178,492 artificially inseminated cows across 9,833 herds in south-western Japan over a 3-year period. The daily mean (±SD) THI over the study period was 63.6 ± 11.3 (range: 41.4-81.5). The calving to first artificial insemination (AI) interval was significantly negatively correlated with THI in the month of AI (r = -.75, p < .001), whereas conception rate to first AI (CRFA) showed a cyclical change that did not correspond with that of THI, resulting in a time lag between CRFA and THI in the month of AI. Furthermore, there was a stronger correlation between CRFA and THI between 3 and 2 months before AI (r = .76 vs. .83, p < .001) than in the month of AI (r = .34, p = .04). Therefore, this extensive field study showed that a decrease in THI was associated with declined reproductive performance in Japanese Black cattle and that the impact of the cold environment on the conception rate is attributable to a carryover effect from the cold season before AI rather than conditions at the time of AI. © 2017 Blackwell Verlag GmbH.
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Aliaga, Leonardo; Lai, Chen; Yu, Jia; Chub, Nikolai; Shim, Hoon; Sun, Lixin; Xie, Chengsong; Yang, Wan-Jou; Lin, Xian; O'Donovan, Michael J.; Cai, Huaibin
2013-01-01
The substitution of Proline with Serine at residue 56 (P56S) of vesicle-associated membrane protein-associated protein B (VAPB) has been linked to an atypical autosomal dominant form of familial amyotrophic lateral sclerosis 8 (ALS8). To investigate the pathogenic mechanism of P56S VAPB in ALS, we generated transgenic (Tg) mice that heterologously express human wild-type (WT) and P56S VAPB under the control of a pan-neuronal promoter Thy1.2. While WT VAPB Tg mice did not exhibit any overt motor behavioral phenotypes, P56S VAPB Tg mice developed progressive hyperactivities and other motor abnormalities. VAPB protein was accumulated as large punctate in the soma and proximal dendrites of both corticospinal motor neurons (CSMNs) and spinal motor neurons (SMNs) in P56S VAPB Tg mice. Concomitantly, a significant increase of endoplasmic reticulum stress and unfolded protein response and the resulting up-regulation of pro-apoptotic factor CCAAT/enhancer-binding protein homologous protein expression were observed in the CSMNs and SMNs of P56S VAPB Tg mice. However, only a progressive loss of CSMNs but not SMNs was found in P56S VAPB Tg mice. In SMNs, P56S VAPB promoted a rather selective translocation of VAPB protein onto the postsynaptic site of C-boutons that altered the morphology of C-boutons and impaired the spontaneous rhythmic discharges of SMNs. Therefore, these findings provide new pathophysiological mechanisms of P56S VAPB that differentially affect the function and survival of CSMNs and SMNs in ALS8. PMID:23771029
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Increased sphingosine-1-phosphate improves muscle regeneration in acutely injured mdx mice
2013-01-01
Background Presently, there is no effective treatment for the lethal muscle wasting disease Duchenne muscular dystrophy (DMD). Here we show that increased sphingosine-1-phoshate (S1P) through direct injection or via the administration of the small molecule 2-acetyl-4(5)-tetrahydroxybutyl imidazole (THI), an S1P lyase inhibitor, has beneficial effects in acutely injured dystrophic muscles of mdx mice. Methods We treated mdx mice with and without acute injury and characterized the histopathological and functional effects of increasing S1P levels. We also tested exogenous and direct administration of S1P on mdx muscles to examine the molecular pathways under which S1P promotes regeneration in dystrophic muscles. Results Short-term treatment with THI significantly increased muscle fiber size and extensor digitorum longus (EDL) muscle specific force in acutely injured mdx limb muscles. In addition, the accumulation of fibrosis and fat deposition, hallmarks of DMD pathology and impaired muscle regeneration, were lower in the injured muscles of THI-treated mdx mice. Furthermore, increased muscle force was observed in uninjured EDL muscles with a longer-term treatment of THI. Such regenerative effects were linked to the response of myogenic cells, since intramuscular injection of S1P increased the number of Myf5nlacz/+ positive myogenic cells and newly regenerated myofibers in injured mdx muscles. Intramuscular injection of biotinylated-S1P localized to muscle fibers, including newly regenerated fibers, which also stained positive for S1P receptor 1 (S1PR1). Importantly, plasma membrane and perinuclear localization of phosphorylated S1PR1 was observed in regenerating muscle fibers of mdx muscles. Intramuscular increases of S1P levels, S1PR1 and phosphorylated ribosomal protein S6 (P-rpS6), and elevated EDL muscle specific force, suggest S1P promoted the upregulation of anabolic pathways that mediate skeletal muscle mass and function. Conclusions These data show that S1P is beneficial for muscle regeneration and functional gain in dystrophic mice, and that THI, or other pharmacological agents that raise S1P levels systemically, may be developed into an effective treatment for improving muscle function and reducing the pathology of DMD. PMID:23915702
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[Determination of genetic bases of auxotrophy in Yersinia pestis ssp. caucasica strains].
Odinokov, G N; Eroshenko, G A; Kukleva, L M; Shavina, N Iu; Krasnov, Ia M; Kutyrev, V V
2012-04-01
Based on the results of computer analysis of nucleotide sequences in strains Yersinia pestis and Y. pseudotuberculosis recorded in the files of NCBI GenBank database, differences between genes argA, aroG, aroF, thiH, and thiG of strain Pestoides F (subspecies caucasica) were found, compared to other strains of plaque agent and pseudotuberculosis microbe. Using PCR with calculated primers and the method of sequence analysis, the structure of variable regions of these genes was studied in 96 natural Y. pestis and Y. pseudotuberculosis strains. It was shown that all examined strains of subspecies caucasica, unlike strains of plague-causing agent of other subspecies and pseudotubercolosis microbe, had identical mutations in genes argA (integration of the insertion sequence IS100), aroG (insertion of ten nucleotides), aroF (inserion of IS100), thiH (insertion of nucleotide T), and thiG (deletion of 13 nucleotides). These mutations are the reason for the absence in strains belonging to this subspecies of the ability to synthesize arginine, phenylalanine, tyrosine, and vitamin B1 (thiamine), and cause their auxotrophy for these growth factors.
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Non-selectivity of new bradykinin antagonists for B1 receptors.
Rhaleb, N E; Gobeil, F; Regoli, D
1992-01-01
Two new B1 receptor antagonists, [Hyp3,Thi5,DTic7,Oic8]desArg9-BK and DArg[Hyp3,Thi5,DTic7,Oic8]desArg9-BK were tested in vitro on the rabbit jugular vein and the guinea pig ileum (preparations containing B2 receptors) and on the rabbit aorta (preparation containing B1 receptors) for pharmacological characterization. The results indicate that both compounds are antagonists on both B1 and B2 receptors, are competitive and discriminate between B2A and B2B receptor subtypes.
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Mollica, Richard F; Chernoff, Miriam C; Megan Berthold, S; Lavelle, James; Lyoo, In Kyoon; Renshaw, Perry
2014-10-01
Little is known about the relationship between traumatic head injury (THI) and psychiatric morbidity in torture survivors. We examine the relationship between THI and depression, PTSD, post-concussive syndrome (PCS), disability and poor health status in Vietnamese ex-political detainees who survived incarceration in Vietnamese re-education camps. A community sample of ex-political detainees (n=337) and a non-THI, non-ex-detainee comparison group (n=82) were surveyed. Seventy-eight percent of the ex-political detainees had experienced THI; 90.6% of the ex-political detainees and 3.6% of the comparison group had experienced 7 or more trauma events. Depression and PTSD were greater in ex-detainees than in the comparison group (40.9% vs 23.2% and 13.4% vs 0%). Dose-effect relationships for THI and trauma/torture in the ex-political detainee group were significant. Logistic regression in the pooled sample of ex-detainees and the comparison group confirmed the independent impact of THI from trauma/torture on psychiatric morbidity (OR for PTSD=22.4; 95% CI: 3.0-165.8). These results demonstrate important effects of THI on depression and PTSD in Vietnamese ex-detainees who have survived torture. Copyright © 2014 Elsevier Inc. All rights reserved.
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NASA Astrophysics Data System (ADS)
Drebes, Julia; Künz, Madeleine; Windshügel, Björn; Kikhney, Alexey G.; Müller, Ingrid B.; Eberle, Raphael J.; Oberthür, Dominik; Cang, Huaixing; Svergun, Dmitri I.; Perbandt, Markus; Betzel, Christian; Wrenger, Carsten
2016-03-01
Infections caused by the methicillin-resistant Staphylococcus aureus (MRSA) are today known to be a substantial threat for global health. Emerging multi-drug resistant bacteria have created a substantial need to identify and discover new drug targets and to develop novel strategies to treat bacterial infections. A promising and so far untapped antibiotic target is the biosynthesis of vitamin B1 (thiamin). Thiamin in its activated form, thiamin pyrophosphate, is an essential co-factor for all organisms. Therefore, thiamin analogous compounds, when introduced into the vitamin B1 biosynthetic pathway and further converted into non-functional co-factors by the bacterium can function as pro-drugs which thus block various co-factor dependent pathways. We characterized one of the key enzymes within the S. aureus vitamin B1 biosynthetic pathway, 5-(hydroxyethyl)-4-methylthiazole kinase (SaThiM; EC 2.7.1.50), a potential target for pro-drug compounds and analyzed the native structure of SaThiM and complexes with the natural substrate 5-(hydroxyethyl)-4-methylthiazole (THZ) and two selected substrate analogues.
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Ruiz-Reig, Nuria; Andres, Belen; Lamonerie, Thomas; Theil, Thomas; Fairén, Alfonso; Studer, Michèle
2018-06-04
In rodents, the medial nucleus of the amygdala receives direct inputs from the accessory olfactory bulbs and is mainly implicated in pheromone-mediated reproductive and defensive behaviors. The principal neurons of the medial amygdala are GABAergic neurons generated principally in the caudo-ventral medial ganglionic eminence and preoptic area. Beside GABAergic neurons, the medial amygdala also contains glutamatergic Otp-expressing neurons cells generated in the lateral hypothalamic neuroepithelium and a non-well characterized Pax6-positive population. In the present work, we describe a novel glutamatergic Ebf3-expressing neuronal subpopulation distributed within the periphery of the postero-ventral medial amygdala. These neurons are generated in a pallial domain characterized by high expression of Gdf10. This territory is topologically the most caudal tier of the ventral pallium and accordingly, we named it Caudo-Ventral Pallium (CVP). In the absence of Pax6, the CVP is disrupted and Ebf3-expressing neurons fail to be generated. Overall, this work proposes a novel model of the neuronal composition of the medial amygdala and unravels for the first time a new novel pallial subpopulation originating from the CVP and expressing the transcription factor Ebf3.
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Bordin, Antonella; Ponti, Donatella; Iudicone, Paola; Rendina, Erino Angelo; Calogero, Antonella; Pierelli, Luca; Ibrahim, Mohsen; De Falco, Elena
2015-01-01
Human adipose tissue-derived mesenchymal stem cells (ADMSCs) are considered eligible candidates for cardiovascular stem cell therapy applications due to their cardiac transdifferentiation potential and immunotolerance. Over the years, the in vitro culture of ADMSCs by platelet lysate (PL), a hemoderivate containing numerous growth factors and cytokines derived from platelet pools, has allowed achieving a safe and reproducible methodology to obtain high cell yield prior to clinical administration. Nevertheless, the biological properties of PL are still to be fully elucidated. In this brief report we show the potential ability of PL to induce a permissive state of cardiac-like transdifferentiation and to cause epigenetic modifications. RTPCR results indicate an upregulation of Cx43, SMA, c-kit, and Thy-1 confirmed by immunofluorescence staining, compared to standard cultures with foetal bovine serum. Moreover, PL-cultured ADMSCs exhibit a remarkable increase of both acetylated histones 3 and 4, with a patient-dependent time trend, and methylation at lysine 9 on histone 3 preceding the acetylation. Expression levels of p300 and SIRT-1, two major regulators of histone 3, are also upregulated after treatment with PL. In conclusion, PL could unravel novel biological properties beyond its routine employment in noncardiac applications, providing new insights into the plasticity of human ADMSCs. PMID:26495284
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Siciliano, Camilla; Chimenti, Isotta; Bordin, Antonella; Ponti, Donatella; Iudicone, Paola; Peruzzi, Mariangela; Rendina, Erino Angelo; Calogero, Antonella; Pierelli, Luca; Ibrahim, Mohsen; De Falco, Elena
2015-01-01
Human adipose tissue-derived mesenchymal stem cells (ADMSCs) are considered eligible candidates for cardiovascular stem cell therapy applications due to their cardiac transdifferentiation potential and immunotolerance. Over the years, the in vitro culture of ADMSCs by platelet lysate (PL), a hemoderivate containing numerous growth factors and cytokines derived from platelet pools, has allowed achieving a safe and reproducible methodology to obtain high cell yield prior to clinical administration. Nevertheless, the biological properties of PL are still to be fully elucidated. In this brief report we show the potential ability of PL to induce a permissive state of cardiac-like transdifferentiation and to cause epigenetic modifications. RTPCR results indicate an upregulation of Cx43, SMA, c-kit, and Thy-1 confirmed by immunofluorescence staining, compared to standard cultures with foetal bovine serum. Moreover, PL-cultured ADMSCs exhibit a remarkable increase of both acetylated histones 3 and 4, with a patient-dependent time trend, and methylation at lysine 9 on histone 3 preceding the acetylation. Expression levels of p300 and SIRT-1, two major regulators of histone 3, are also upregulated after treatment with PL. In conclusion, PL could unravel novel biological properties beyond its routine employment in noncardiac applications, providing new insights into the plasticity of human ADMSCs.
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Westerweel, Peter E.; Joles, Jaap A.; den Ouden, Krista; Goldschmeding, Roel; Rookmaaker, Maarten B.; Verhaar, Marianne C.
2012-01-01
Background/Aims ACE inhibitor (ACE-I) treatment effectively inhibits proteinuria and ameliorates the course of various renal diseases. In experimental glomerulonephritis, however, angiotensin II (AngII) infusion has also been shown to be renoprotective. We evaluated the long-term (28 days) course of anti-Thy1 glomerulonephritis in animals with suppressed AngII formation by ACE-I treatment. Methods Brown Norway rats received perindopril (2.8 mg/kg/day, n = 12), dihydropyridine calcium-antagonist amlodipine (Ca-A; 13 mg/kg/day, n = 6) or were left untreated (n = 14). All animals were monitored for blood pressure, proteinuria, and creatinine clearance after anti-Thy1 injection. Renal histology was assessed at day 7 and 28. Results Systolic blood pressure was equally reduced by ACE-I and Ca-A treatment. AngII suppression prevented development of proteinuria, but did not protect against glomerular microaneurysm formation or reduction in creatinine clearance. After resolution of the microaneurysms, animals with suppressed AngII production showed a modest increase in glomerulosclerosis and vasculopathic thickening of intrarenal vessels. Conclusions In anti-Thy1 glomerulonephritis, suppression of AngII formation does not protect against the induction of glomerular damage and is associated with mild aggravation of adverse renal fibrotic remodeling. Proteinuria, however, is effectively prevented by ACE-I treatment. Ca-A treatment did not affect the course of glomerulonephritis, indicating that ACE-I effects are blood pressure independent. PMID:22479264
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Ketteler, M; Westenfeld, R; Gawlik, A; de Heer, E; Distler, A
2000-01-01
Pathways of L-arginine metabolism including nitric oxide, agmatine and polyamine synthesis are upregulated during glomerular inflammation in experimental glomerulonephritis. In anti-Thy-1-glomerulonephritis L-arginine-deficient diets ameliorate the disease course in this model. However, it is unclear which metabolic pathway is affected by this substrate depletion. Since polyamines are important proproliferative molecules, we studied the effect of specific polyamine synthesis blockade in vivo on mesangial cell proliferation and glomerular fibrosis in this model. Anti-Thy-1-glomerulonephritis was induced in male Sprague-Dawley rats by single-bolus injection of monoclonal ER4-antibodies. Rats were treated with difluoromethylornithine (0.5-2% in the drinking water), a selective inhibitor of the rate-limiting enzyme of polyamine synthesis, ornithine decarboxylase (ODC). Mesangial cell proliferation and matrix expansion were evaluated in PAS-stained kidney tissues. Glomerular TGF-beta and biglycan-mRNA-expression were determined by Northern blot analysis and albuminuria was measured using a competitive ELISA. Data were compared to untreated controls. Though complete inhibition of ODC activity was achieved at any time point, difluoromethlornithine treatment had no significant effect on albuminuria, glomerular matrix protein expression and mesangial cell count in this model. The acute upregulation of glomerular ODC activity above baseline in anti-Thy1-glomerulonephritis is not pathophysiologically important for disease development however, biological effects of available polyamine pools cannot be excluded by our study.
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Inhibitory dendrite dynamics as a general feature of the adult cortical microcircuit.
Chen, Jerry L; Flanders, Genevieve H; Lee, Wei-Chung Allen; Lin, Walter C; Nedivi, Elly
2011-08-31
The mammalian neocortex is functionally subdivided into architectonically distinct regions that process various types of information based on their source of afferent input. Yet, the modularity of neocortical organization in terms of cell type and intrinsic circuitry allows afferent drive to continuously reassign cortical map space. New aspects of cortical map plasticity include dynamic turnover of dendritic spines on pyramidal neurons and remodeling of interneuron dendritic arbors. While spine remodeling occurs in multiple cortical regions, it is not yet known whether interneuron dendrite remodeling is common across primary sensory and higher-level cortices. It is also unknown whether, like pyramidal dendrites, inhibitory dendrites respect functional domain boundaries. Given the importance of the inhibitory circuitry to adult cortical plasticity and the reorganization of cortical maps, we sought to address these questions by using two-photon microscopy to monitor interneuron dendritic arbors of thy1-GFP-S transgenic mice expressing GFP in neurons sparsely distributed across the superficial layers of the neocortex. We find that interneuron dendritic branch tip remodeling is a general feature of the adult cortical microcircuit, and that remodeling rates are similar across primary sensory regions of different modalities, but may differ in magnitude between primary sensory versus higher cortical areas. We also show that branch tip remodeling occurs in bursts and respects functional domain boundaries.
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Chitin-Induced Airway Epithelial Cell Innate Immune Responses Are Inhibited by Carvacrol/Thymol
Erle, David J.
2016-01-01
Chitin is produced in large amounts by fungi, insects, and other organisms and has been implicated in the pathogenesis of asthma. Airway epithelial cells are in direct contact with environmental particles and serve as the first line of defense against inhaled allergens and pathogens. The potential contributions of airway epithelial cells to chitin-induced asthma remain poorly understood. We hypothesized that chitin directly stimulates airway epithelial cells to release cytokines that promote type 2 immune responses and to induce expression of molecules which are important in innate immune responses. We found that chitin exposure rapidly induced the expression of three key type 2-promoting cytokines, IL-25, IL-33 and TSLP, in BEAS-2B transformed human bronchial epithelial cells and in A549 and H292 lung carcinoma cells. Chitin also induced the expression of the key pattern recognition receptors TLR2 and TLR4. Chitin induced the expression of miR-155, miR-146a and miR-21, each of which is known to up-regulate the expression of pro-inflammatory cytokines. Also the expression of SOCS1 and SHIP1 which are known targets of miR-155 was repressed by chitin treatment. The monoterpene phenol carvacrol (Car) and its isomer thymol (Thy) are found in herbal essential oils and have been shown to inhibit allergic inflammation in asthma models. We found that Car/Thy inhibited the effects of chitin on type 2-promoting cytokine release and on the expression of TLRs, SOCS1, SHIP1, and miRNAs. Car/Thy could also efficiently reduce the protein levels of TLR4, inhibit the increase in TLR2 protein levels in chitin plus Car/Thy-treated cells and increase the protein levels of SHIP1 and SOCS1, which are negative regulators of TLR-mediated inflammatory responses. We conclude that direct effects of chitin on airway epithelial cells are likely to contribute to allergic airway diseases like asthma, and that Car/Thy directly inhibits epithelial cell pro-inflammatory responses to chitin. PMID:27463381
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Zhang, Peng; Sun, Hongwen; Min, Lujuan; Ren, Chao
2018-05-01
One interest of using biochar as soil amendment is to reduce pesticide adverse effects. In this paper, the sorption and degradation of thiacloprid (THI) in a black soil amended by various biochars were systematically investigated, and the mechanisms therein were explored by analyzing the changes in soil physicochemical properties, degrading enzymes and genes and microorganism community. Biochar amendment increased THI sorption in soil, which was associated with an increase in organic carbon and surface area and a decrease in H/C. Amendments of 300-PT (pyrolyzing temperature) biochar promoted the biodegradation of THI by increasing the microbe abundance and improving nitrile hydratase (NHase) activity. In contrast, 500- and 700-PT biochar amendments inhibited biodegradation by reducing THI availability and changing NHase activity and THI-degradative nth gene abundance, and instead promoted chemical degradation mainly through elevated pH, active groups on mineral surface and generation of •OH and other free radicals. Furthermore, THI shifted the soil microbial community, stimulated the NHase activity and elevated nth gene abundance. Biochar amendments also changed soil bacterial community by modulating soil pH, dissolved organic matter and nitrogen and phosphorus levels, which further influenced THI biodegradation. Therefore, the impact of biochars on the fate of a pesticide in soil depends greatly on their type and properties, which should be comprehensively examined when applying biochar to soil. Copyright © 2018 Elsevier Ltd. All rights reserved.
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Skarzynski, Piotr H; Raj-Koziak, Danuta; J Rajchel, Joanna; Pilka, Adam; Wlodarczyk, Andrzej W; Skarzynski, Henryk
2017-10-01
To describe how the Tinnitus Handicap Inventory (THI) was translated into Polish (THI-POL) and to present psychometric data on how well it performed in a clinical population of tinnitus sufferers. The original version of THI was adapted into Polish. The reliability of THI-POL was investigated using test-retest, Cronbach's alpha, endorsement rate and item-total correlation. Construct validity and convergent validity were also assessed based on confirmatory factor analysis, inter-item correlation and Pearson product-moment correlations using subscale A (Tinnitus) of the Tinnitus and Hearing Survey (THS-POL); divergent validity was checked using subscale B (Hearing) of THS-POL. A group of 167 adults filled in THI-POL twice over their three-day hospitalisation period. Test-retest reliability for the total THI-POL scores was strong (r = 0.91). Cronbach's alpha coefficient for the total score was high (r = 0.95), confirming the questionnaire's stability. Confirmatory factor analysis (CFA) and inter-item correlation did not confirm the three-factor model. Convergent validity from the Tinnitus subscale of THS showed a positive strong (r = 0.75) correlation. Divergent validity showed only a moderate correlation. All analyses were statistically significant (p < 0.01). THI-POL is a valid and reliable self-administered tool, which allows the overall tinnitus handicap of Polish-speaking patients to be effectively assessed.
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A de novo compound targeting α-synuclein improves deficits in models of Parkinson’s disease
Wrasidlo, Wolfgang; Tsigelny, Igor F.; Price, Diana L.; Dutta, Garima; Rockenstein, Edward; Schwarz, Thomas C.; Ledolter, Karin; Bonhaus, Douglas; Paulino, Amy; Eleuteri, Simona; Skjevik, Åge A.; Kouznetsova, Valentina L.; Spencer, Brian; Desplats, Paula; Gonzalez-Ruelas, Tania; Trejo-Morales, Margarita; Overk, Cassia R.; Winter, Stefan; Zhu, Chunni; Chesselet, Marie-Francoise; Meier, Dieter; Moessler, Herbert; Konrat, Robert; Masliah, Eliezer
2016-01-01
Abnormal accumulation and propagation of the neuronal protein α-synuclein has been hypothesized to underlie the pathogenesis of Parkinson’s disease, dementia with Lewy bodies and multiple system atrophy. Here we report a de novo-developed compound (NPT100-18A) that reduces α-synuclein toxicity through a novel mechanism that involves displacing α-synuclein from the membrane. This compound interacts with a domain in the C-terminus of α-synuclein. The E83R mutation reduces the compound interaction with the 80–90 amino acid region of α-synuclein and prevents the effects of NPT100-18A. In vitro studies showed that NPT100-18A reduced the formation of wild-type α-synuclein oligomers in membranes, reduced the neuronal accumulation of α-synuclein, and decreased markers of cell toxicity. In vivo studies were conducted in three different α-synuclein transgenic rodent models. Treatment with NPT100-18A ameliorated motor deficits in mThy1 wild-type α-synuclein transgenic mice in a dose-dependent manner at two independent institutions. Neuropathological examination showed that NPT100-18A decreased the accumulation of proteinase K-resistant α-synuclein aggregates in the CNS and was accompanied by the normalization of neuronal and inflammatory markers. These results were confirmed in a mutant line of α-synuclein transgenic mice that is prone to generate oligomers. In vivo imaging studies of α-synuclein-GFP transgenic mice using two-photon microscopy showed that NPT100-18A reduced the cortical synaptic accumulation of α-synuclein within 1 h post-administration. Taken together, these studies support the notion that altering the interaction of α-synuclein with the membrane might be a feasible therapeutic approach for developing new disease-modifying treatments of Parkinson’s disease and other synucleinopathies. PMID:27679481
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Lee, Hyun-Jin; Kim, Min-Beom; Yoo, Shin-Young; Park, Shi Nae; Nam, Eui-Cheol; Moon, In Seok; Lee, Ho-Ki
2018-01-01
The purpose of this study was to investigate the effectiveness of intratympanic dexamethasone injection (ITDI) in acute tinnitus of presumed cochlear origin. A prospective, randomized, placebo-controlled, double-blinded, multicenter study. Between August 2013 and December 2015, 54 patients with unilateral tinnitus were enrolled at four different centers. Patients were assigned either to an ITDI (n = 27) or an intratympanic normal saline injection (ITNI; n = 27) group through block randomization. Intratympanic injections were administered four times over 2 weeks. At 4 weeks after initial injection, we analyzed the improvement rates of tinnitus using the tinnitus handicap Inventory (THI) and visual analogue scale (VAS) for loudness, awareness, and annoyance. We defined improvement as the reduction of more than 7 points or of more than 20% in the final THI score compared to the initial THI score. The initial mean hearing thresholds and VAS and THI scores of the two groups did not differ significantly. At 4 weeks after initial injection, the mean VAS and THI scores of both groups had significantly reduced. However, the improvement rate did not differ significantly between the groups (ITDI, 51.9%; ITNI, 59.3%). The results indicate that ITDI might not be more effective than ITNI for the treatment of acute unilateral tinnitus. Therefore, ITDI should not be considered as the main treatment for patients presenting with acute tinnitus as the primary symptom. 1b. Laryngoscope, 128:184-188, 2018. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.
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El-Tarabany, Mahmoud Salah
2016-07-01
The aim of this study was to investigate the effect of temperature-humidity index (THI) level on productive parameters, welfare, and immunity in Japanese quails. One hundred and eighty (180) birds of Japanese quail, 14 weeks old, were used. Birds were divided randomly into three equal groups, control (at low THI, less than 70), H1 (at moderate THI, 70-75), and H2 (at high THI, 76-80). Birds in the control group had higher body weight (281.2 g, p = 0.001), egg mass (745 g, p = 0.001), fertility (85.4 %, p = 0.039), hatchability (80.4 %, p = 0.001), and immune response titer to Newcastle disease virus (p = 0.031), compared with H2 group. Furthermore, the thermoneutral group had higher internal egg quality score [albumen height (5.14 mm, p = 0.001), yolk height (10.88 mm, p = 0.015), yolk index (42.32 %, p = 0.039), and Haugh unit (92.67, p = 0.001)]. Nevertheless, there were no significant differences in fertility percentage, immune response, and corticosterone concentration between control and H1 group. Birds in the H2 group had the lowest total leucocytic count and lymphocyte percentage (p = 0.001 and 0.020, respectively) but the highest H/L ratio (0.83, p = 0.001). Corticosterone concentration was lower in control and H1 groups (5.49 and 6.41 ng/mL, respectively, p = 0.024) than that in H2 group. Japanese quail exposed to heat stress revealed drop in production and immunological parameters, as well as a detrimental effects on welfare. Thus, practical approaches might be used to reduce the detrimental effects of greater THI level.
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Carter, B H; Friend, T H; Garey, S M; Sawyer, J A; Alexander, M B; Tomazewski, M A
2014-01-01
The objective of this study was to determine the ability of radiant insulation hutch covers to moderate the effect of ambient temperature and radiant energy on calves housed in polyethylene hutches. The insulation had a double layer of polyethylene bubble film laminated between a layer of aluminum foil and white polyethylene (reflectance = 95%, R value (ft(2) · °F ·h/Btu) = 2.7). In each of two experiments (exp.), hutches were either uninsulated (control) or covered with reflective insulation across the top and sides of the hutch leaving the front, back, and pen exposed. Each hutch had a 1.2 × 1.8-m attached outdoor wire pen. In both exp., rate of increase of interior hutch temperature relative to ambient temperature was lower in insulated hutches (P < 0.001) indicating they were warmer at low THI and cooler at high THI. In exp. 1, increase in respiration rate and ear canal temperature of the calves, relative to THI, were moderated in insulated hutches (P < 0.001). In Exp. 2, respiration rate was not affected by treatment (P = 0.50), but increased with increasing THI (P < 0.001). Mean ADG did not differ among treatments in either exp. (P > 0.21). Insulating calf hutches with reflective insulation moderated hutch microclimate, and improved calf comfort, but did not translate to improvements in economically relevant variables such as ADG.
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Danaei, Gholam Hassan; Karami, Mohammad
2017-10-01
Several studies have shown that oxidative stress and cell damage can occur in the very early stages of diazinon (DZN) exposure. The present study was designed to determine the beneficial effect of thymoquinone (Thy), the main component of Nigella sativa (black seed or black cumin) against DZN immunotoxicity, hematotoxicity and genotoxicity in rats. In the present experimental study, 48 male Wistar rats were randomly divided into six groups, (eight per group) as follows: control (receiving corn oil as the DZN solvent), DZN (20mg/kg), Thy (10mg/kg), Thy (2.5mg/kg)+DZN, Thy (5mg/kg)+DZN and Thy (10mg/kg)+DZN. After four weeks of treatment, the hematological parameters of red blood cells (RBCs), white blood cells (WBCs), hemoglobin (Hb), hematocrit (Hct) and platelets (PLTs) were evaluated. The evaluation of genotoxicity was carried out using the micronucleus assay. For measurement of cytokine production, interferon gamma (IFN-γ), interleukin 10 (IL10) and interleukin 4 (IL4) were chosen as immunotoxicity indicators of DZN toxicity. DZN was found to decrease RBCs, WBCs, Hb, Hct, PLTs, butyrl- and acetyl-cholinesterase activity and I FN-γ and increased the micronucleus indices of IL10 and IL4 as compared with the control group. Treatment with Thy reduced DZN hematotoxicity and immunotoxicity, but, significantly, did not prevent genotoxicity. This study showed that Thy (without the significant effect on genotoxicity) decreased the hematological toxicity, immunotoxicity and butyrl and acetyl cholinesterase activity induced by DZN. The success of Thy supplementation against DZN toxicity can be attributed to the antioxidant effects of its constituents. Copyright © 2017. Published by Elsevier B.V.
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Arduini, Agnese; Redaelli, Veronica; Luzi, Fabio; Dall'Olio, Stefania; Pace, Vincenzo; Nanni Costa, Leonardo
2017-02-10
In order to evaluate the relationships between deck level, body surface temperature and carcass damages after a short journey (30 min), 10 deliveries of Italian heavy pigs, including a total of 1400 animals from one farm, were examined. Within 5 min after the arrival at the abattoir, the vehicles were unloaded. Environmental temperature and relative humidity were recorded and a Temperature Humidity Index (THI) was calculated. After unloading, maximum temperatures of dorsal and ocular regions were measured by a thermal camera on groups of pigs from each of the unloaded decks. After dehairing, quarters and whole carcasses were evaluated subjectively by a trained operator for skin damage using a four-point scale. On the basis of THI at unloading, deliveries were grouped into three classes. Data of body surface temperature and skin damage score were analysed in a model including THI class, deck level and their interaction. Regardless of pig location in the truck, the maximum temperature of the dorsal and ocular regions increased with increasing THI class. Within each THI class, the highest and lowest body surface temperatures were found in pigs located on the middle and upper decks, respectively. Only THI class was found to affect the skin damage score ( p < 0.05), which increased on quarters and whole carcasses with increasing THI class. The results of this study on short-distance transport of Italian heavy pigs highlighted the need to control and ameliorate the environmental conditions in the trucks, even at relatively low temperature and THI, in order to improve welfare and reduce loss of carcass value.
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Macroscopic in vivo imaging of facial nerve regeneration in Thy1-GFP rats.
Placheta, Eva; Wood, Matthew D; Lafontaine, Christine; Frey, Manfred; Gordon, Tessa; Borschel, Gregory H
2015-01-01
Facial nerve injury leads to severe functional and aesthetic deficits. The transgenic Thy1-GFP rat is a new model for facial nerve injury and reconstruction research that will help improve clinical outcomes through translational facial nerve injury research. To determine whether serial in vivo imaging of nerve regeneration in the transgenic rat model is possible, facial nerve regeneration was imaged under the main paradigms of facial nerve injury and reconstruction. Fifteen male Thy1-GFP rats, which express green fluorescent protein (GFP) in their neural structures, were divided into 3 groups in the laboratory: crush-injury, direct repair, and cross-face nerve grafting (30-mm graft length). The distal nerve stump or nerve graft was predegenerated for 2 weeks. The facial nerve of the transgenic rats was serially imaged at the time of operation and after 2, 4, and 8 weeks of regeneration. The imaging was performed under a GFP-MDS-96/BN excitation stand (BLS Ltd). Facial nerve injury. Optical fluorescence of regenerating facial nerve axons. Serial in vivo imaging of the regeneration of GFP-positive axons in the Thy1-GFP rat model is possible. All animals survived the short imaging procedures well, and nerve regeneration was followed over clinically relevant distances. The predegeneration of the distal nerve stump or the cross-face nerve graft was, however, necessary to image the regeneration front at early time points. Crush injury was not suitable to sufficiently predegenerate the nerve (and to allow for degradation of the GFP through Wallerian degeneration). After direct repair, axons regenerated over the coaptation site in between 2 and 4 weeks. The GFP-positive nerve fibers reached the distal end of the 30-mm-long cross-face nervegrafts after 4 to 8 weeks of regeneration. The time course of facial nerve regeneration was studied by serial in vivo imaging in the transgenic rat model. Nerve regeneration was followed over clinically relevant distances in a small number of experimental animals, as they were subsequently imaged at multiple time points. The Thy1-GFP rat model will help improve clinical outcomes of facial reanimation surgery through improving the knowledge of facial nerve regeneration after surgical procedures. NA.
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Krolicki, Leszek; Bruchertseifer, Frank; Kunikowska, Jolanta; Koziara, Henryk; Królicki, Bartosz; Jakuciński, Maciej; Pawlak, Dariusz; Apostolidis, Christos; Mirzadeh, Saed; Rola, Rafał; Merlo, Adrian; Morgenstern, Alfred
2018-04-30
Glioblastoma multiforme (GBM), the most common malignant brain tumor, mainly manifests as a primary de novo and less frequently as a secondary glial neoplasm. GBM has been demonstrated to overexpress the NK-1 receptor and substance P can be used as a ligand for targeted therapy. Alpha emitters, e.g. 213 Bi, that deposit their high energy within a short range allow the selective irradiation of tumor cells while sparing adjacent neuronal structures. Among 50 glioma patients of different subtypes that have to date been treated with targeted alpha therapy at the Medical University Warsaw, we report here the data on nine patients with secondary GBM. Following surgery, chemo- and radiotherapy, recurrent GBM was treated by intracavitary injection of 1-6 doses of 0.9-2.3 GBq 213 Bi- DOTA-[Thi 8 ,Met(O 2 ) 11 ]-substance P ( 213 Bi-DOTA-SP) in 2-month intervals. 68 Ga-DOTA-[Thi 8 ,Met(O 2 ) 11 ]-substance P ( 68 Ga-DOTA-SP) was co-injected with the therapeutic doses to assess biodistribution using PET/CT. Therapeutic response was monitored with MRI. Treatment with activities ranging from 1.4 to 9.7 (median 5.8) GBq 213 Bi- DOTA-SP was well tolerated with only mild transient adverse reactions, mainly headaches due to a transient perfocal edema reaction. The median progression free survival and overall survival time following the initiation of alpha therapy was 5.8 and 16.4 months, respectively. The median overall survival time from the first diagnosis was 52.3 months. Two out of nine patients are still alive 39 and 51 months, respectively, after the initiation of the therapy. Targeted alpha therapy of secondary GBM with 213 Bi-DOTA-SP is safe and well tolerated and may evolve as a promising novel therapeutic option for secondary GBM.
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The Influence of Vegetation Function towards the Langsep Street Thermal Comfort
NASA Astrophysics Data System (ADS)
Alfian, R.; Setyabudi, I.; Uran, R. S.
2017-10-01
Streetscape is an important element for character building of the environment, spatial, and visual in order to provide an urban identity, especially in Malang City protocol streets. Langsep Street is one of the protocol streets in Malang City. Langsep Street famous with central education and offices area. This study aims (1) to identify vegetation of streetscape; (2) to analyze the thermal comfort of the streetscape, and (3) to evaluate the comfort level of Langsep Street. The method used was the THI approach. THI value that obtained was analyzed using the standard of Laurie (1990). Based on observations, the THI value of Langsep Street was 27.60. This was influenced by the trees canopy density and spacing of the trees on the streetscape. It can be concluded that streetscape required (1) shaded plants that have root systems that do not damage the construction of roads, (2) the branching plants are not easily broken and easy to maintain, and (3) the combination of trees, shrubs and ground cover.
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Chamberland, Simon; Yang, Helen H; Pan, Michael M; Evans, Stephen W; Guan, Sihui; Chavarha, Mariya; Yang, Ying; Salesse, Charleen; Wu, Haodi; Wu, Joseph C; Clandinin, Thomas R; Toth, Katalin; Lin, Michael Z; St-Pierre, François
2017-07-27
Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in Drosophila . These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision.
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Metabolomics of human brain aging and age-related neurodegenerative diseases.
Jové, Mariona; Portero-Otín, Manuel; Naudí, Alba; Ferrer, Isidre; Pamplona, Reinald
2014-07-01
Neurons in the mature human central nervous system (CNS) perform a wide range of motor, sensory, regulatory, behavioral, and cognitive functions. Such diverse functional output requires a great diversity of CNS neuronal and non-neuronal populations. Metabolomics encompasses the study of the complete set of metabolites/low-molecular-weight intermediates (metabolome), which are context-dependent and vary according to the physiology, developmental state, or pathologic state of the cell, tissue, organ, or organism. Therefore, the use of metabolomics can help to unravel the diversity-and to disclose the specificity-of metabolic traits and their alterations in the brain and in fluids such as cerebrospinal fluid and plasma, thus helping to uncover potential biomarkers of aging and neurodegenerative diseases. Here, we review the current applications of metabolomics in studies of CNS aging and certain age-related neurodegenerative diseases such as Alzheimer disease, Parkinson disease, and amyotrophic lateral sclerosis. Neurometabolomics will increase knowledge of the physiologic and pathologic functions of neural cells and will place the concept of selective neuronal vulnerability in a metabolic context.
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Pang, Zhen; Wu, Nancy N; Zhao, Weiguang; Chain, David C; Schaffer, Erica; Zhang, Xin; Yamdagni, Preeti; Palejwala, Vaseem A; Fan, Chunpeng; Favara, Sarah G; Dressler, Holly M; Economides, Kyriakos D; Weinstock, Daniel; Cavallo, Jean S; Naimi, Souad; Galzin, Anne-Marie; Guillot, Etienne; Pruniaux, Marie-Pierre; Tocci, Michael J; Polites, H Greg
2011-10-01
Cannabinoid receptor CB1 is expressed abundantly in the brain and presumably in the peripheral tissues responsible for energy metabolism. It is unclear if the antiobesity effects of rimonabant, a CB1 antagonist, are mediated through the central or the peripheral CB1 receptors. To address this question, we generated transgenic mice with central nervous system (CNS)-specific knockdown (KD) of CB1, by expressing an artificial microRNA (AMIR) under the control of the neuronal Thy1.2 promoter. In the mutant mice, CB1 expression was reduced in the brain and spinal cord, whereas no change was observed in the superior cervical ganglia (SCG), sympathetic trunk, enteric nervous system, and pancreatic ganglia. In contrast to the neuronal tissues, CB1 was undetectable in the brown adipose tissue (BAT) or the liver. Consistent with the selective loss of central CB1, agonist-induced hypothermia was attenuated in the mutant mice, but the agonist-induced delay of gastrointestinal transit (GIT), a primarily peripheral nervous system-mediated effect, was not. Compared to wild-type (WT) littermates, the mutant mice displayed reduced body weight (BW), adiposity, and feeding efficiency, and when fed a high-fat diet (HFD), showed decreased plasma insulin, leptin, cholesterol, and triglyceride levels, and elevated adiponectin levels. Furthermore, the therapeutic effects of rimonabant on food intake (FI), BW, and serum parameters were markedly reduced and correlated with the degree of CB1 KD. Thus, KD of CB1 in the CNS recapitulates the metabolic phenotype of CB1 knockout (KO) mice and diminishes rimonabant's efficacy, indicating that blockade of central CB1 is required for rimonabant's antiobesity actions.
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Hill, D L; Wall, E
2015-01-01
A better understanding of how livestock respond to weather is essential to enable farming to adapt to a changing climate. Climate change is mainly expected to impact dairy cattle through heat stress and an increase in the frequency of extreme weather events. We investigated the effects of weather on milk yield and composition (fat and protein content) in an experimental dairy herd in Scotland over 21 years. Holstein Friesian cows were either housed indoors in winter and grazed over the summer or were continuously housed. Milk yield was measured daily, resulting in 762 786 test day records from 1369 individuals, and fat and protein percentage were sampled once a week, giving 89 331 records from 1220 cows/trait. The relative influence of 11 weather elements, measured from local outdoor weather stations, and two indices of temperature and humidity (THI), indicators of heat stress, were compared using separate maximum likelihood models for each element or index. Models containing a direct measure of temperature (dry bulb, wet bulb, grass or soil temperature) or a THI provided the best fits to milk yield and fat data; wind speed and the number of hours of sunshine were most important in explaining protein content. Weather elements summarised across a week's timescale from the test day usually explained milk yield and fat content better than shorter-scale (3 day, test day, test day -1) metrics. Then, examining a subset of key weather variables using restricted maximum likelihood, we found that THI, wind speed and the number of hours of sunshine influenced milk yield and composition. The shape and magnitude of these effects depended on whether animals were inside or outside on the test day. The milk yield of cows outdoors was lower at the extremes of THI than at average values, and the highest yields were obtained when THI, recorded at 0900 h, was 55 units. Cows indoors decreased milk yield as THI increased. Fat content was lower at higher THIs than at intermediate THIs in both environments. Protein content decreased as THI increased in animals kept indoors and outdoors, and the rate of decrease was greater when animals were outside than when they were inside. Moderate wind speeds appeared to alleviate heat stress. These results show that milk yield and composition are impacted at the upper extreme of THI under conditions currently experienced in Scotland, where animals have so far experienced little pressure to adapt to heat stress.
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NASA Astrophysics Data System (ADS)
Alamiddine, Zakaria; Selvam, Balaji; Cerón-Carrasco, José P.; Mathé-Allainmat, Monique; Lebreton, Jacques; Thany, Steeve H.; Laurent, Adèle D.; Graton, Jérôme; Le Questel, Jean-Yves
2015-12-01
The binding of thiaclopride (THI), a neonicotinoid insecticide, with Aplysia californica acetylcholine binding protein ( Ac-AChBP), the surrogate of the extracellular domain of insects nicotinic acetylcholine receptors, has been studied with a QM/QM' hybrid methodology using the ONIOM approach (M06-2X/6-311G(d):PM6). The contributions of Ac-AChBP key residues for THI binding are accurately quantified from a structural and energetic point of view. The importance of water mediated hydrogen-bond (H-bond) interactions involving two water molecules and Tyr55 and Ser189 residues in the vicinity of the THI nitrile group, is specially highlighted. A larger stabilization energy is obtained with the THI- Ac-AChBP complex compared to imidacloprid (IMI), the forerunner of neonicotinoid insecticides. Pairwise interaction energy calculations rationalize this result with, in particular, a significantly more important contribution of the pivotal aromatic residues Trp147 and Tyr188 with THI through CH···π/CH···O and π-π stacking interactions, respectively. These trends are confirmed through a complementary non-covalent interaction (NCI) analysis of selected THI- Ac-AChBP amino acid pairs.
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Gillard, Geoffrey O.; Bivas-Benita, Maytal; Hovav, Avi-Hai; Grandpre, Lauren E.; Panas, Michael W.; Seaman, Michael S.; Haynes, Barton F.; Letvin, Norman L.
2011-01-01
While immunological memory has long been considered the province of T- and B- lymphocytes, it has recently been reported that innate cell populations are capable of mediating memory responses. We now show that an innate memory immune response is generated in mice following infection with vaccinia virus, a poxvirus for which no cognate germline-encoded receptor has been identified. This immune response results in viral clearance in the absence of classical adaptive T and B lymphocyte populations, and is mediated by a Thy1+ subset of natural killer (NK) cells. We demonstrate that immune protection against infection from a lethal dose of virus can be adoptively transferred with memory hepatic Thy1+ NK cells that were primed with live virus. Our results also indicate that, like classical immunological memory, stronger innate memory responses form in response to priming with live virus than a highly attenuated vector. These results demonstrate that a defined innate memory cell population alone can provide host protection against a lethal systemic infection through viral clearance. PMID:21829360
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Arshad, Q; Roberts, R E; Ahmad, H; Lobo, R; Patel, M; Ham, T; Sharp, D J; Seemungal, B M
2017-04-01
We hypothesised that chronic vestibular symptoms (CVS) of imbalance and dizziness post-traumatic head injury (THI) may relate to: (i) the occurrence of multiple simultaneous vestibular diagnoses including both peripheral and central vestibular dysfunction in individual patients increasing the chance of missed diagnoses and suboptimal treatment; (ii) an impaired response to vestibular rehabilitation since the central mechanisms that mediate rehabilitation related brain plasticity may themselves be disrupted. We report the results of a retrospective analysis of both the comprehensive clinical and vestibular laboratory testing of 20 consecutive THI patients with prominent and persisting vestibular symptoms still present at least 6months post THI. Individual THI patients typically had multiple vestibular diagnoses and unique to this group of vestibular patients, often displayed both peripheral and central vestibular dysfunction. Despite expert neuro-otological management, at two years 20% of patients still had persisting vestibular symptoms. In summary, chronic vestibular dysfunction in THI could relate to: (i) the presence of multiple vestibular diagnoses, increasing the risk of 'missed' vestibular diagnoses leading to persisting symptoms; (ii) the impact of brain trauma which may impair brain plasticity mediated repair mechanisms. Apart from alerting physicians to the potential for multiple vestibular diagnoses in THI, future work to identify the specific deficits in brain function mediating poor recovery from post-THI vestibular dysfunction could provide the rationale for developing new therapy for head injury patients whose vestibular symptoms are resistant to treatment. Copyright © 2017. Published by Elsevier B.V.
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Walsh, Carolyn M.; Bautista, Diana M.; Lumpkin, Ellen A.
2015-01-01
An assortment of touch receptors innervate the skin and encode different tactile features of the environment. Compared with invertebrate touch and other sensory systems, our understanding of the molecular and cellular underpinnings of mammalian touch lags behind. Two recent breakthroughs have accelerated progress. First, an arsenal of cell-type-specific molecular markers allowed the functional and anatomical properties of sensory neurons to be matched, thereby unraveling a cellular code for touch. Such markers have also revealed key roles of non-neuronal cell types, such as Merkel cells and keratinocytes, in touch reception. Second, the discovery of Piezo genes as a new family of mechanically activated channels has fueled the discovery of molecular mechanisms that mediate and mechanotransduction in mammalian touch receptors. PMID:26100741
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1980-01-01
Heavily irradiated peritoneal cells (PC) from congenitally athymic nude (nu/nu) mice markedly restored the impaired in vitro antibody response of nu/nu spleen cells to sheep erythrocyte antigens (T-dependent antigen), whereas irradiated spleen or lymph node cells from nu/nu mice had no effect on the response. This activity of the irradiated PC of nu/nu mice was completely abolished by treatment with anti-Thy-1.2 antiserum plus normal guinea pig serum (C') and is, therefore, attributable to a function of matured T cells. PMID:6966314
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Santana, M L; Pereira, R J; Bignardi, A B; Filho, A E Vercesi; Menéndez-Buxadera, A; El Faro, L
2015-12-01
In an attempt to determine the possible detrimental effects of continuous selection for milk yield on the genetic tolerance of Zebu cattle to heat stress, genetic parameters and trends of the response to heat stress for 86,950 test-day (TD) milk yield records from 14,670 first lactations of purebred dairy Gir cows were estimated. A random regression model with regression on days in milk (DIM) and temperature-humidity index (THI) values was applied to the data. The most detrimental effect of THI on milk yield was observed in the stage of lactation with higher milk production, DIM 61 to 120 (-0.099kg/d per THI). Although modest variations were observed for the THI scale, a reduction in additive genetic variance as well as in permanent environmental and residual variance was observed with increasing THI values. The heritability estimates showed a slight increase with increasing THI values for any DIM. The correlations between additive genetic effects across the THI scale showed that, for most of the THI values, genotype by environment interactions due to heat stress were less important for the ranking of bulls. However, for extreme THI values, this type of genotype by environment interaction may lead to an important error in selection. As a result of the selection for milk yield practiced in the dairy Gir population for 3 decades, the genetic trend of cumulative milk yield was significantly positive for production in both high (51.81kg/yr) and low THI values (78.48kg/yr). However, the difference between the breeding values of animals at high and low THI may be considered alarming (355kg in 2011). The genetic trends observed for the regression coefficients related to general production level (intercept of the reaction norm) and specific ability to respond to heat stress (slope of the reaction norm) indicate that the dairy Gir population is heading toward a higher production level at the expense of lower tolerance to heat stress. These trends reflect the genetic antagonism between production and tolerance to heat stress demonstrated by the negative genetic correlation between these components (-0.23). Monitoring trends of the genetic component of heat stress would be a reasonable measure to avoid deterioration in one of the main traits of Zebu cattle (i.e., high tolerance to heat stress). On the basis of current genetic trends, the need for future genetic evaluation of dairy Zebu animals for tolerance to heat stress cannot be ruled out. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
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A Complete Developmental Sequence of a Drosophila Neuronal Lineage as Revealed by Twin-Spot MARCM
He, Yisheng; Ding, Peng; Kao, Jui-Chun; Lee, Tzumin
2010-01-01
Drosophila brains contain numerous neurons that form complex circuits. These neurons are derived in stereotyped patterns from a fixed number of progenitors, called neuroblasts, and identifying individual neurons made by a neuroblast facilitates the reconstruction of neural circuits. An improved MARCM (mosaic analysis with a repressible cell marker) technique, called twin-spot MARCM, allows one to label the sister clones derived from a common progenitor simultaneously in different colors. It enables identification of every single neuron in an extended neuronal lineage based on the order of neuron birth. Here we report the first example, to our knowledge, of complete lineage analysis among neurons derived from a common neuroblast that relay olfactory information from the antennal lobe (AL) to higher brain centers. By identifying the sequentially derived neurons, we found that the neuroblast serially makes 40 types of AL projection neurons (PNs). During embryogenesis, one PN with multi-glomerular innervation and 18 uniglomerular PNs targeting 17 glomeruli of the adult AL are born. Many more PNs of 22 additional types, including four types of polyglomerular PNs, derive after the neuroblast resumes dividing in early larvae. Although different offspring are generated in a rather arbitrary sequence, the birth order strictly dictates the fate of each post-mitotic neuron, including the fate of programmed cell death. Notably, the embryonic progenitor has an altered temporal identity following each self-renewing asymmetric cell division. After larval hatching, the same progenitor produces multiple neurons for each cell type, but the number of neurons for each type is tightly regulated. These observations substantiate the origin-dependent specification of neuron types. Sequencing neuronal lineages will not only unravel how a complex brain develops but also permit systematic identification of neuron types for detailed structure and function analysis of the brain. PMID:20808769
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Li, Xueyan; Kan, Xianwen
2018-04-30
In this study, a ratiometric strategy-based electrochemical sensor was developed by electropolymerization of thionine (THI) and β-cyclodextrin (β-CD) composite films on a glassy carbon electrode surface for imidacloprid (IMI) detection. THI played the role of an inner reference element to provide a built-in correction. In addition, the modified β-CD showed good selective enrichment for IMI to improve the sensitivity and anti-interference ability of the sensor. The current ratio between IMI and THI was calculated as the detected signal for IMI sensing. Compared with common single-signal sensing, the proposed ratiometric strategy showed a higher linear range and a lower limit of detection of 4.0 × 10-8-1.0 × 10-5 mol L-1 and 1.7 × 10-8 mol L-1, respectively, for IMI detection. On the other hand, the ratiometric strategy endowed the sensor with good accuracy, reproducibility, and stability. The sensor was also used for IMI determination in real samples with satisfactory results. The simple, effective, and reliable way reported in this study can be further used to prepare ratiometric strategy-based electrochemical sensors for the selective and sensitive detection of other compounds with good accuracy and stability.
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Goldhawk, C; Crowe, T; Janzen, E; González, L A; Kastelic, J; Pajor, E; Schwartzkopf-Genswein, K S
2014-11-01
Nineteen loads of commercial feeder cattle (BW 376 ± 39 kg, mean ± SD) transported for 18 ± 4.5 h in summer and winter seasons were used to collect data on internal temperature and humidity conditions in the deck and belly compartment of pot-bellied trailers and their relationship with shrink, cortisol, and morbidity. Measurements of temperature or humidity at ceiling or animal level did not vary with transportation factors. Temperature and humidity ratio was greater at animal-level than ambient conditions during nonhighway travel and stationary periods (P < 0.01). During the 3 time periods evaluated within journeys, there was a larger difference between animal-level and ambient conditions during the winter than during the summer (P < 0.01); however, this difference was not associated with other transport factors (P > 0.05). Evening loads (1700 and 2100 h) experienced more shrink in the summer than in the winter (11.2 ± 0.5 vs. 9.0 ± 0.5% of BW; P = 0.03). A 1°C increase in difference between average animal-level temperature in transit and the mean ambient temperature during the 10 d before transport was associated with a 0.11 ± 0.03% of BW increase in shrink (P < 0.01) and 0.006 ± 0.002 ng/mL increase in posttransport cortisol concentration (P = 0.05). Animal-level temperature-humidity index (THI) events (consecutive observations of THI greater than 78°F) were more likely to last for longer than 1 h when the trailer was stationary vs. traveling (mean = 1.8, confidence level 95% = 1.33, 2.52). During THI events at animal level, the disagreement with ambient temperature regarding THI classification was lower when the vehicle was traveling vs. stationary (95.5 ± 0.01% vs. 99.7 ± 0.002% of THI event in disagreement; P < 0.01) and was greatest in events less than 1 h (99.8 ± 0.0% vs. 91.7 ± 0.03% of THI event in disagreement; P < 0.01). The average magnitude of the difference during these events was 11.4 ± 7.6°F and was not affected by transportation factors (P > 0.05). Despite association between indicators of calf welfare and microclimate, all cattle arrived in good condition and there was 0.96% treatment rate within the first 30 d after arrival. Management and auditing decisions related to transportation of feeder cattle should consider the relationship between animal-level and ambient conditions and conditions before transportation. Under the commercial conditions of the current study, the transportation process did not appear to cause distress according to the dimensions of animal welfare that were assessed.
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Sun, Yu-Yo; Li, Yikun; Wali, Bushra; Li, Yuancheng; Lee, Jolly; Heinmiller, Andrew; Abe, Koji; Stein, Donald G.; Mao, Hui; Sayeed, Iqbal; Kuan, Chia-Yi
2015-01-01
Background and Purpose Hypoperfusion-induced thrombosis is an important mechanism for post-surgery stroke and cognitive decline, but there are no perioperative neuroprotectants to date. This study investigated whether prophylactic application of Edaravone, a free radical scavenger already used in treating ischemic stroke in Japan, can prevent infarct and cognitive deficits in a murine model of transient cerebral hypoxia-ischemia. Methods Adult male C57BL/6 mice were subjected to transient hypoxic-ischemic (tHI) insult that consists of 30-min occlusion of the unilateral common carotid artery and exposure to 7.5% oxygen. Edaravone or saline was prophylactically applied to compare their effects on cortical oxygen saturation, blood flow, coagulation, oxidative stress, metabolites, and learning-memory using methods that include photoacoustic imaging, laser speckle contrast imaging, solid state NMR and Morris water maze. The effects on infarct size by Edaravone application at different time-points after tHI were also compared. Results Prophylactic administration of Edaravone (4.5 mg/kg × 2, IP, 1 h before and 1 h after tHI) improved vascular reperfusion, oxygen saturation, and the maintenance of brain metabolites, while reducing oxidative stress, thrombosis, white-matter injury, and learning impairment after tHI insult. Delayed Edaravone treatment after 3 h post-tHI became unable to reduce infarct size. Conclusions Acute application of Edaravone may be a useful strategy to prevent post-surgery stroke and cognitive impairment, especially in patients with severe carotid stenosis. PMID:26060244
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Sun, Yu-Yo; Li, Yikun; Wali, Bushra; Li, Yuancheng; Lee, Jolly; Heinmiller, Andrew; Abe, Koji; Stein, Donald G; Mao, Hui; Sayeed, Iqbal; Kuan, Chia-Yi
2015-07-01
Hypoperfusion-induced thrombosis is an important mechanism for postsurgery stroke and cognitive decline, but there are no perioperative neuroprotectants to date. This study investigated whether prophylactic application of Edaravone, a free radical scavenger already used in treating ischemic stroke in Japan, can prevent infarct and cognitive deficits in a murine model of transient cerebral hypoxia-ischemia. Adult male C57BL/6 mice were subjected to transient hypoxic-ischemic (tHI) insult that consists of 30-minute occlusion of the unilateral common carotid artery and exposure to 7.5% oxygen. Edaravone or saline was prophylactically applied to compare their effects on cortical oxygen saturation, blood flow, coagulation, oxidative stress, metabolites, and learning-memory using methods that include photoacoustic imaging, laser speckle contrast imaging, solid-state NMR, and Morris water maze. The effects on infarct size by Edaravone application at different time points after tHI were also compared. Prophylactic administration of Edaravone (4.5 mg/kg×2, IP, 1 hour before and 1 hour after tHI) improved vascular reperfusion, oxygen saturation, and the maintenance of brain metabolites, reducing oxidative stress, thrombosis, white-matter injury, and learning impairment after tHI insult. Delayed Edaravone treatment after 3 h post-tHI became unable to reduce infarct size. Acute application of Edaravone may be a useful strategy to prevent postsurgery stroke and cognitive impairment, especially in patients with severe carotid stenosis. © 2015 American Heart Association, Inc.
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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Transmyocardial Laser Revascularization
... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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NASA Astrophysics Data System (ADS)
Santolaria, Pilar; López-Gatius, Fernando; García-Ispierto, Irina; Bech-Sàbat, Gregori; Angulo, Eduardo; Carretero, Teresa; Sánchez-Nadal, Jóse Antonio; Yániz, Jesus
2010-01-01
The aim of this study was to determine possible relationships between farm climate conditions, recorded from day 0 to day 40 post-artificial insemination (AI), and late embryo/early fetal loss in high producing dairy cows. Pregnancy was diagnosed by rectal ultrasonography between 28 and 34 days post-AI. Fetal loss was registered when a further 80- to 86-day diagnosis proved negative. Climate variables such as air temperature and relative humidity (RH) were monitored in the cubicles area for each 30-min period. Temperature-humidity indices (THI); cumulative stressful values and episodes of acute change (defined as the mean daily value 1.2 times higher or lower than the mean daily values of the 10 previous days) of the climate variables were calculated. The data were derived from 759 cows in one herd. A total of 692 pregnancies (91.2%) carried singletons and 67 (8.8%) carried twins. No triplets were recorded. Pregnancy loss was recorded in 6.7% (51/759) of pregnancies: 5.6% (39/692) in single and 17.9% (12/67) in twin pregnancies. Using logistic regression procedures, a one-unit increase in the daily cumulative number of hours for the THI values higher than 85 during days 11-20 of gestation caused a 1.57-fold increase in the pregnancy loss, whereas the likelihood of fetal loss increased by a factor of 1.16 for each additional episode of acute variation for the maximum THI values during gestation days 0-40. THI values higher than 85 and episodes of acute variation for the maximum THI values were only recorded during the warm and cool periods, respectively. The presence of twins led to a 3.98-fold increase in pregnancy loss. In conclusion, our findings show that cumulative stressful and episodes of acute variation of climatic conditions can compromise the success of gestation during both the cool and warm periods of the year. Twin pregnancy was confirmed as a main factor associated with pregnancy loss.
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Rodriguez, Guillaume; Sarazin, Matthieu; Clemente, Alexandra; Holden, Stephanie; Paz, Jeanne T; Delord, Bruno
2018-04-30
Persistent neural activity, the substrate of working memory, is thought to emerge from synaptic reverberation within recurrent networks. However, reverberation models do not robustly explain fundamental dynamics of persistent activity, including high-spiking irregularity, large intertrial variability, and state transitions. While cellular bistability may contribute to persistent activity, its rigidity appears incompatible with persistent activity labile characteristics. Here, we unravel in a cellular model a form of spike-mediated conditional bistability that is robust, generic and provides a rich repertoire of mnemonic computations. Under asynchronous synaptic inputs of the awakened state, conditional bistability generates spiking/bursting episodes, accounting for the irregularity, variability and state transitions characterizing persistent activity. This mechanism has likely been overlooked because of the sub-threshold input it requires and we predict how to assess it experimentally. Our results suggest a reexamination of the role of intrinsic properties in the collective network dynamics responsible for flexible working memory. SIGNIFICANCE STATEMENT This study unravels a novel form of intrinsic neuronal property, i.e. conditional bistability. We show that, thanks of its conditional character, conditional bistability favors the emergence of flexible and robust forms of persistent activity in PFC neural networks, in opposition to previously studied classical forms of absolute bistability. Specifically, we demonstrate for the first time that conditional bistability 1) is a generic biophysical spike-dependent mechanism of layer V pyramidal neurons in the PFC and that 2) it accounts for essential neurodynamical features for the organisation and flexibility of PFC persistent activity (the large irregularity and intertrial variability of the discharge and its organization under discrete stable states), which remain unexplained in a robust fashion by current models. Copyright © 2018 the authors.
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Marchetti, B
1996-01-01
Neurons and astrocytes have a close anatomic and functional relationship that plays a crucial role during development and in the adult brain. Astrocytes in the central nervous system (CNS) express receptors for a variety of growth factors (GFs), neurotransmitters and/or neuromodulators; in turn, neuronal cells can respond to astrocyte-derived GFs and control astrocyte function via a common set of signaling molecules and intracellular transducing pathways. There is also increasing evidence that soluble factors from lymphoid/mononuclear cells are able to modulate the growth and function of cells found in the CNS, specifically macroglial and microglial cells. Furthermore, glial cells can secrete immunoregulatory molecules that influence immune cells as well as the glial cells themselves. As neuronal and immune cells share common signaling systems, the potential exists for bidirectional communication not only between lymphoid and glial cells, but also between neuronal cells and immune and glial cells. In the present work, interactions of luteinizing-hormone-releasing hormone (LHRH) and the astroglial cell are proposed as a prototype for the study of neuroimmune communication within the CNS in the light of (1) the commonality of signal molecules (hormones, neurotransmitters and cytokines) and transduction mechanisms shared by glia LHRH neurons and lymphoid cells; (2) the central role of glia in the developmental organization and pattern of LHRH neuronal migration during embryogenesis, and (3) the strong modulatory role played by sex steroids in mechanisms involved in synaptic and interneuronal organization, as well as in the sexual dimorphisms of neuroendocrine-immune functions. During their maturation and differentiation in vitro, astroglial cells release factors able to accelerate markedly the LHRH neuronal phenotypic differentiation as well as the acquisition of mature LHRH secretory potential, with a potency depending on both the 'age' and the specific brain localization of the astroglia, as well as the degree of LHRH neuronal differentiation in vitro. Regional differences in astroglial sensitivity to estrogens were also measured. Different experimental paradigms such as coculture and mixed-culture models between the immortalized LHRH (GT1-1) neuronal cell line and astroglial cells in primary culture, disclosed the presence of a bidirectional flow of informational molecules regulating both proliferative and secretory capacities of each cell type. The importance of adhesive mechanisms in such cross-talk is underscored by the complete abolition of GT1-1 LHRH production and cell proliferation following the counteraction of neuronal-neuronal/neuronal-glial interactions through addition of neural-cell adhesion molecule antiserum. Other information came from pharmacological experiments manipulating the astroglia-derived cytokines and/or nitric oxide, which revealed cross-talk between the neuronal and astroglial compartments. From the bulk of this information, it seems likely that interactions between astroglia and LHRH neurons play a major role in the integration of the multiplicity of brain signals converging on the LHRH neurons that govern reproduction. Another important facet of neuronal-glial interactions is that concerning neuron-guided migration, and unraveling astroglial/LHRH-neuronal networks might then constitute an additional effort in the comprehension of defective LHRH-neuronal migration in Kallman's syndrome.
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Deng, Keqin; Liu, Xinyan; Li, Chunxiang; Huang, Haowen
2018-05-31
The loading capacity of thionin (Thi) on shortened multi-walled carbon nanotubes (S-MWCNTs) and acidified multi-walled carbon nanotubes (A-MWCNTs) was compared. Two DNA probe fragments were designed for hybridization with microRNA-21 (miR-21), the microRNAs (miRNAs) model analyte. DNA probe 1 (P1) was assembled on Au nanoparticles (AuNPs) modified electrode. MiR-21 was captured by the pre-immobilized P1. A signal nanoprobe was synthesized by loading large amount of Thi on S-MWCNTs with covalently bonded probe 2 (P2). Owing to the large effective surface area of MWCNTs, fast electron shuttle of MWCNTs, high-loaded Thi on S-MWCNTs, and the increased conductivity from AuNPs, after signal probe hybridized with miR-21, it gave rise to a magnified current response on electrode. The increased electrochemical current enabled us to quantitatively detect miR-21. Expensive bioreagents and labeled target/detection DNA or miRNAs were avoided in this strategy. The operation complexity and assay cost were also reduced. Copyright © 2018 Elsevier B.V. All rights reserved.
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Chung, Wilson C J; Linscott, Megan L; Rodriguez, Karla M; Stewart, Courtney E
2016-01-01
Over the last few years, numerous studies solidified the hypothesis that fibroblast growth factor (FGF) signaling regulates neuroendocrine progenitor cell proliferation, fate specification, and cell survival and, therefore, is critical for the regulation and maintenance of homeostasis of the body. One important example that underscores the involvement of FGF signaling during neuroendocrine cell development is gonadotropin-releasing hormone (GnRH) neuron ontogenesis. Indeed, transgenic mice with reduced olfactory placode (OP) Fgf8 expression do not have GnRH neurons. This observation indicates the requirement of FGF8 signaling for the emergence of the GnRH neuronal system in the embryonic OP, the putative birth place of GnRH neurons. Mammalian reproductive success depends on the presence of GnRH neurons to stimulate gonadotropin secretion from the anterior pituitary, which activates gonadal steroidogenesis and gametogenesis. Together, these observations are critical for understanding the function of GnRH neurons and their control of the hypothalamus-pituitary-gonadal (HPG) axis to maintain fertility. Taken together, these studies illustrate that GnRH neuron emergence and hence HPG function is vulnerable to genomic and molecular signals that abnormally modify Fgf8 expression in the developing mouse OP. In this short review, we focus on research that is aimed at unraveling how androgen, all-trans retinoic acid, and how epigenetic factors modify control mouse OP Fgf8 transcription in the context of GnRH neuronal development and mammalian reproductive success.
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Inoue, Takeshi; Yamashita, Taiga; Agata, Kiyokazu
2014-11-19
For most organisms, sensitive recognition of even slight changes in environmental temperature is essential for adjusting their behavioral strategies to ensure homeostasis and survival. However, much remains to be understood about the molecular and cellular processes that regulate thermosensation and the corresponding behavioral responses. Planarians display clear thermotaxis, although they have a relatively simple brain. Here, we devised a quantitative thermotaxis assay and unraveled a neural pathway involved in planarian thermotaxis by combinatory behavioral assays and RNAi analysis. We found that thermosensory neurons that expressed a planarian Dugesia japonica homolog of the Transient Receptor Potential Melastatin family a (DjTRPMa) gene were required for the thermotaxis. Interestingly, although these thermosensory neurons are distributed throughout their body, planarians with a dysfunctional brain due to regeneration-dependent conditional gene knockdown (Readyknock) of the synaptotagmin gene completely lost their thermotactic behavior. These results suggest that brain function is required as a central processor for the thermosensory response. Therefore, we investigated the type(s) of brain neurons involved in processing the thermal signals by gene knockdown of limiting enzymes for neurotransmitter biosynthesis in the brain. We found that serotonergic neurons with dendrites that were elongated toward DjTRPMa-expressing thermosensory neurons might be required for the processing of signals from thermosensory neurons that results in thermotaxis. These results suggest that serotonergic neurons in the brain may interact with thermosensory neurons activated by TRPM ion channels to produce thermotaxis in planarians. Copyright © 2014 the authors 0270-6474/14/3415701-14$15.00/0.
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Diagnostic Tests and Procedures
... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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Testing of Streptococcus pneumoniae for resistance to penicillin.
Marshall, K J; Musher, D M; Watson, D; Mason, E O
1993-01-01
The increasing prevalence of penicillin-resistant Streptococcus pneumoniae requires antibiotic susceptibility tests that can be done with greater ease and reliability. We measured the MIC of penicillin for pneumococci by the tube macrodilution method with Mueller-Hinton broth (MHB), Haemophilus Test Medium (HTM), Todd-Hewitt broth with 0.5% yeast extract (THY), and MHB with 3% lysed horse blood (LHB). Eight (19%) and 6 (14%) of 42 pneumococcal isolates failed to generate turbid growth in MHB and HTM, respectively, whereas all pneumococcal isolates did so in THY and LHB. For those strains that replicated to turbidity, the mean MICs of penicillin were lower in MHB and HTM than in THY and LHB, with differences being significant (P < 0.05) for comparisons with LHB. Four isolates appeared to be penicillin susceptible in HTM but were actually moderately resistant in THY and LHB, and two isolates appeared to be moderately resistant but were resistant. A similar failure to detect resistance was seen with MHB. S. pneumoniae ATCC 49619, a moderately penicillin-resistant strain that has been proposed for quality control testing, gave variable results in MHB or THM and appeared to be susceptible to penicillin in some assays, whereas the MICs for S. pneumoniae ATCC 49619 in THY or LHB fell within a twofold dilution range, with geometric means of 0.16 and 0.18 micrograms/ml, respectively. Pneumococcal isolates thus may appear falsely susceptible to penicillin when tested in MHB or HTM. LHB remains the standard medium; however, because THY is an easily prepared clear medium that can be used in automated systems and appears to yield results similar to those obtained with LHB, THY deserves consideration for routine use. PMID:8501225
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Base Realignment and Closure: An Evolution for Harvesting Efficiencies
2012-06-15
Master’s Thesis 3. DATES COVERED (From - To) 25-07-2011 to 15-06-2012 4. TITLE AND SUBTITLE Base Realignment and Closure: An Evolution for...made to c lo~e and rea l ign bases in a timely manner. ACKNOWLEDGEl\\IIENT Although one name appears on the cover of thi.s thesi ~. thi~ paper i the...wisdom to endure to the end. • M y fami ly ror your constcu1t encourugcment and support . • C’ A PT Joanne Fish. my thesi ~ advi!’>ur who provided the
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Input transformation by dendritic spines of pyramidal neurons
Araya, Roberto
2014-01-01
In the mammalian brain, most inputs received by a neuron are formed on the dendritic tree. In the neocortex, the dendrites of pyramidal neurons are covered by thousands of tiny protrusions known as dendritic spines, which are the major recipient sites for excitatory synaptic information in the brain. Their peculiar morphology, with a small head connected to the dendritic shaft by a slender neck, has inspired decades of theoretical and more recently experimental work in an attempt to understand how excitatory synaptic inputs are processed, stored and integrated in pyramidal neurons. Advances in electrophysiological, optical and genetic tools are now enabling us to unravel the biophysical and molecular mechanisms controlling spine function in health and disease. Here I highlight relevant findings, challenges and hypotheses on spine function, with an emphasis on the electrical properties of spines and on how these affect the storage and integration of excitatory synaptic inputs in pyramidal neurons. In an attempt to make sense of the published data, I propose that the raison d'etre for dendritic spines lies in their ability to undergo activity-dependent structural and molecular changes that can modify synaptic strength, and hence alter the gain of the linearly integrated sub-threshold depolarizations in pyramidal neuron dendrites before the generation of a dendritic spike. PMID:25520626
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Diseases of the Tricuspid Valve
... Daily Conferences Fellowships and Residencies School of Perfusion Technology Education Resources Library & Learning Resource Center CME Resources THI Journal THI Cardiac Society Save the Date: International Symposium on Cardiovascular Regenerative ...
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Fu, X; Xu, J G
2000-01-01
A chromosome-plasmid balanced lethal gene delivery system for Lactobacillus acidophilus based on the thyA gene was developed. The selected L. acidophilus DOM La strain carries a mutated thyA gene and has an obligate requirement for thymidine. This strain can be used as a host for the constructed shuttle vector pFXL03, lacking antibiotic-resistant markers but having the wild-type thyA gene from L. casei which complements the thyA chromosomal mutation. The vector also contains the replicon region from plasmid pUC19 and that of the Lactococcus plasmid pWV01, which allows the transfer between Escherichia coli, L. casei and L. acidophilus. Eight unique restriction sites (i.e., PstI, HindIII, SphI, SalI, AccI, XbaI, KpnI and SacI) are available for cloning. After 40-time transfers in modified MRS medium, no plasmid loss was observed. The vector pFXL03 is potentially useful as a food-grade vaccine delivery system for L. acidophilus.
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Adaptation of the Arabic Version of the Tinnitus Handicap Inventory.
Barake, Rana; Rizk, Samer Abou; Ziade, Georges; Zaytoun, George; Bassim, Marc
2016-03-01
To translate the Tinnitus Handicap Inventory (THI) into literary Arabic to come up with a unified Arabic version and to determine its validity and reliability in assessing the quality of life of Arabic-speaking patients with tinnitus. Clinical measurement study. Tertiary care center. The original English THI was translated into literary Arabic by a forward- and back-translation process according to the published guidelines for cross-cultural adaptation of health-related quality-of-life measures and applied to 100 patients with chronic tinnitus. Internal consistency reliability was then assessed by calculating Cronbach's alpha coefficient. Pearson correlation coefficients were also calculated for the different scales and the different baseline characteristics. Results showed high internal consistency and reliability coefficients (total THI: 0.93, functional subscale: 0.86, emotional subscale: 0.86, catastrophic subscale: 0.66) comparable to those of the original English THI. The Arabic version of the THI is a valid and reliable tool for the assessment of the impact of tinnitus on the quality of life of Arabic-speaking patients with the complaint of chronic tinnitus. © American Academy of Otolaryngology—Head and Neck Surgery Foundation 2015.
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Chamberland, Simon; Yang, Helen H; Pan, Michael M; Evans, Stephen W; Guan, Sihui; Chavarha, Mariya; Yang, Ying; Salesse, Charleen; Wu, Haodi; Wu, Joseph C; Clandinin, Thomas R; Toth, Katalin; Lin, Michael Z; St-Pierre, François
2017-01-01
Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in Drosophila. These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision. DOI: http://dx.doi.org/10.7554/eLife.25690.001 PMID:28749338
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Endonuclease G mediates α-synuclein cytotoxicity during Parkinson's disease.
Büttner, Sabrina; Habernig, Lukas; Broeskamp, Filomena; Ruli, Doris; Vögtle, F Nora; Vlachos, Manolis; Macchi, Francesca; Küttner, Victoria; Carmona-Gutierrez, Didac; Eisenberg, Tobias; Ring, Julia; Markaki, Maria; Taskin, Asli Aras; Benke, Stefan; Ruckenstuhl, Christoph; Braun, Ralf; Van den Haute, Chris; Bammens, Tine; van der Perren, Anke; Fröhlich, Kai-Uwe; Winderickx, Joris; Kroemer, Guido; Baekelandt, Veerle; Tavernarakis, Nektarios; Kovacs, Gabor G; Dengjel, Jörn; Meisinger, Chris; Sigrist, Stephan J; Madeo, Frank
2013-11-27
Malfunctioning of the protein α-synuclein is critically involved in the demise of dopaminergic neurons relevant to Parkinson's disease. Nonetheless, the precise mechanisms explaining this pathogenic neuronal cell death remain elusive. Endonuclease G (EndoG) is a mitochondrially localized nuclease that triggers DNA degradation and cell death upon translocation from mitochondria to the nucleus. Here, we show that EndoG displays cytotoxic nuclear localization in dopaminergic neurons of human Parkinson-diseased patients, while EndoG depletion largely reduces α-synuclein-induced cell death in human neuroblastoma cells. Xenogenic expression of human α-synuclein in yeast cells triggers mitochondria-nuclear translocation of EndoG and EndoG-mediated DNA degradation through a mechanism that requires a functional kynurenine pathway and the permeability transition pore. In nematodes and flies, EndoG is essential for the α-synuclein-driven degeneration of dopaminergic neurons. Moreover, the locomotion and survival of α-synuclein-expressing flies is compromised, but reinstalled by parallel depletion of EndoG. In sum, we unravel a phylogenetically conserved pathway that involves EndoG as a critical downstream executor of α-synuclein cytotoxicity.
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Kaniuka-Jakubowska, Sonia; Lewczuk, Anna; Majkowicz, Mikołaj; Piskunowicz, Maciej; Mizan-Gross, Krystyna; Zapaśnik, Adam; Kaszubowski, Mariusz; Lass, Piotr; Sworczak, Krzysztof
2018-01-01
Despite numerous publications regarding nontoxic goiter (NTG) treatment and an increasing interest in patients' quality of life, few studies present the outcome of 131-I treatment from the patients' perspective. Our study's main aim was to verify whether there is any improvement in life quality following 131-I treatment. Thirty-five patients with NTG qualified to participate in the study. All patients completed a Thyroid-Related Health-Related Quality of Life (Thy-R-HRQoL) questionnaire created by us and the Medical Outcomes Study 36-item Short Form (SF-36), right before and 1 year after 131-I. We observed an improvement in six out of eight SF-36 and three out of seven Thy-R-HRQoL domains. In comparison with the control group, we observed worse results in two out of eight, prior to treatment, and one out of eight SF-36 afterward, as well as in all Thy-R-HRQoL domains. We did not find any correlation between improvement of Thy-R-HRQoL and SF-36 and goiter size reduction, except for Bodily Pain. There was also no correlation between improvement of SF-36 and Thy-R-HRQoL domains, and goiter size before treatment. The older the patient, the less noticeable improvement was observed in Physical and Social Functioning, and Vitality in SF-36, but age had no influence on the assessment by Thy-R-HRQoL. Radioiodine treatment improves life quality in patients with NTG. Use of the Health-Related Quality of Life questionnaire should be taken into consideration when evaluating life quality of patients with NTG. Relentless pursuit of maximal goiter size reduction in 131-I treatment is worth consideration. In our study, life quality improvement did not depend directly on the goiter size reduction. Life quality improvement after 131-I might not depend on initial goiter size, and for certain domains of SF-36 might be less clearly expressed in older patients.
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Boekhoudt, Linde; Wijbrans, Ellen C; Man, Jodie H K; Luijendijk, Mieneke C M; de Jong, Johannes W; van der Plasse, Geoffrey; Vanderschuren, Louk J M J; Adan, Roger A H
2018-01-01
Motivational deficits are a key symptom in multiple psychiatric disorders, including major depressive disorder, schizophrenia and addiction. A likely neural substrate for these motivational deficits is the brain dopamine (DA) system. In particular, DA signalling in the nucleus accumbens, which originates from DA neurons in the ventral tegmental area (VTA), has been identified as a crucial substrate for effort-related and activational aspects of motivation. Unravelling how VTA DA neuronal activity relates to motivational behaviours is required to understand how motivational deficits in psychiatry can be specifically targeted. In this study, we therefore used designer receptors exclusively activated by designer drugs (DREADD) in TH:Cre rats, in order to determine the effects of chemogenetic DA neuron activation on different aspects of motivational behaviour. We found that chemogenetic activation of DA neurons in the VTA, but not substantia nigra, significantly increased responding for sucrose under a progressive ratio schedule of reinforcement. More specifically, high effort exertion was characterized by increased initiations of reward-seeking actions. This effect was dependent on effort requirements and instrumental contingencies, but was not affected by sucrose pre-feeding. Together, these findings indicate that VTA DA neuronal activation drives motivational behaviour by facilitating action initiation. With this study, we show that enhancing excitability of VTA DA neurons is a viable strategy to improve motivational behaviour. Copyright © 2017 Elsevier B.V. and ECNP. All rights reserved.
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Transgenic mouse lines for non-invasive ratiometric monitoring of intracellular chloride
Batti, Laura; Mukhtarov, Marat; Audero, Enrica; Ivanov, Anton; Paolicelli, Rosa Chiara; Zurborg, Sandra; Gross, Cornelius; Bregestovski, Piotr; Heppenstall, Paul A.
2013-01-01
Chloride is the most abundant physiological anion and participates in a variety of cellular processes including trans-epithelial transport, cell volume regulation, and regulation of electrical excitability. The development of tools to monitor intracellular chloride concentration ([Cli]) is therefore important for the evaluation of cellular function in normal and pathological conditions. Recently, several Cl-sensitive genetically encoded probes have been described which allow for non-invasive monitoring of [Cli]. Here we describe two mouse lines expressing a CFP-YFP-based Cl probe called Cl-Sensor. First, we generated transgenic mice expressing Cl-Sensor under the control of the mouse Thy1 mini promoter. Cl-Sensor exhibited good expression from postnatal day two (P2) in neurons of the hippocampus and cortex, and its level increased strongly during development. Using simultaneous whole-cell monitoring of ionic currents and Cl-dependent fluorescence, we determined that the apparent EC50 for Cli was 46 mM, indicating that this line is appropriate for measuring neuronal [Cli] in postnatal mice. We also describe a transgenic mouse reporter line for Cre-dependent conditional expression of Cl-Sensor, which was targeted to the Rosa26 locus and by incorporating a strong exogenous promoter induced robust expression upon Cre-mediated recombination. We demonstrate high levels of tissue-specific expression in two different Cre-driver lines targeting cells of the myeloid lineage and peripheral sensory neurons. Using these mice the apparent EC50 for Cli was estimated to be 61 and 54 mM in macrophages and DRG, respectively. Our data suggest that these mouse lines will be useful models for ratiometric monitoring of Cli in specific cell types in vivo. PMID:23734096
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Comparison of precipitation measurements by OTT Parsivel2 and Thies LPM optical disdrometers
NASA Astrophysics Data System (ADS)
Angulo-Martínez, Marta; Beguería, Santiago; Latorre, Borja; Fernández-Raga, María
2018-05-01
Optical disdrometers are present weather sensors with the ability of providing detailed information on precipitation such as rain intensity, radar reflectivity or kinetic energy, together with discrete information on the particle size and fall velocity distribution (PSVD) of the hydrometeors. Disdrometers constitute a step forward towards a more complete characterization of precipitation, being useful in several research fields and applications. In this article the performance of two extensively used optical disdrometers, the most recent version of OTT Parsivel2 disdrometer and Thies Clima Laser Precipitation Monitor (LPM), is evaluated. During 2 years, four collocated optical disdrometers, two Thies Clima LPM and two OTT Parsivel2, collected up to 100 000 min of data and up to 30 000 min with rain in more than 200 rainfall events, with intensities peaking at 277 mm h-1 in 1 minute. The analysis of these records shows significant differences between both disdrometer types for all integrated precipitation parameters, which can be explained by differences in the raw PSVD estimated by the two sensors. Thies LPM recorded a larger number of particles than Parsivel2 and a higher proportion of small particles than OTT Parsivel2, resulting in higher rain rates and totals and differences in radar reflectivity and kinetic energy. These differences increased greatly with rainfall intensity. Possible causes of these differences, and their practical consequences, are discussed in order to help researchers and users in the choice of sensor, and at the same time pointing out limitations to be addressed in future studies.
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Yasuyama, Kouji; Meinertzhagen, Ian A
2010-02-01
Recent studies in Drosophila melanogaster indicate that the neuropeptide pigment-dispersing factor (PDF) is an important output signal from a set of major clock neurons, s-LN(v)s (small ventral lateral neurons), which transmit the circadian phase to subsets of other clock neurons, DNs (dorsal neurons). Both s-LN(v)s and DNs have fiber projections to the dorsal protocerebrum of the brain, so that this area is a conspicuous locus for coupling between different subsets of clock neurons. To unravel the neural circuits underlying the fly's circadian rhythms, we examined the detailed subcellular morphology of the PDF-positive fibers of the s-LN(v)s in the dorsal protocerebrum, focusing on their synaptic connections, using preembedding immunoelectron microscopy. To examine the distribution of synapses, we also reconstructed the three-dimensional morphology of PDF-positive varicosities from fiber profiles in the dorsal protocerebrum. The varicosities contained large dense-core vesicles (DCVs), and also numerous small clear vesicles, forming divergent output synapses onto unlabeled neurites. The DCVs apparently dock at nonsynaptic sites, suggesting their nonsynaptic release. In addition, a 3D reconstruction revealed the presence of input synapses onto the PDF-positive fibers. These were detected less frequently than output sites. These observations suggest that the PDF-positive clock neurons receive neural inputs directly through synaptic connections in the dorsal protocerebrum, in addition to supplying dual outputs, either synaptic or via paracrine release of the DCV contents, to unidentified target neurons.
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Trace Elements: What They Do and Where to Get Them
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Kriegeskorte, Andre; Block, Desiree; Drescher, Mike; Windmüller, Nadine; Mellmann, Alexander; Baum, Cathrin; Neumann, Claudia; Lorè, Nicola Ivan; Bragonzi, Alessandra; Liebau, Eva; Hertel, Patrick; Seggewiss, Jochen; Becker, Karsten; Proctor, Richard A.; Peters, Georg
2014-01-01
ABSTRACT Staphylococcus aureus thymidine-dependent small-colony variants (TD-SCVs) are frequently isolated from patients with chronic S. aureus infections after long-term treatment with trimethoprim-sulfamethoxazole (TMP-SMX). While it has been shown that TD-SCVs were associated with mutations in thymidylate synthase (TS; thyA), the impact of such mutations on protein function is lacking. In this study, we showed that mutations in thyA were leading to inactivity of TS proteins, and TS inactivity led to tremendous impact on S. aureus physiology and virulence. Whole DNA microarray analysis of the constructed ΔthyA mutant identified severe alterations compared to the wild type. Important virulence regulators (agr, arlRS, sarA) and major virulence determinants (hla, hlb, sspAB, and geh) were downregulated, while genes important for colonization (fnbA, fnbB, spa, clfB, sdrC, and sdrD) were upregulated. The expression of genes involved in pyrimidine and purine metabolism and nucleotide interconversion changed significantly. NupC was identified as a major nucleoside transporter, which supported growth of the mutant during TMP-SMX exposure by uptake of extracellular thymidine. The ΔthyA mutant was strongly attenuated in virulence models, including a Caenorhabditis elegans killing model and an acute pneumonia mouse model. This study identified inactivation of TS as the molecular basis of clinical TD-SCV and showed that thyA activity has a major role for S. aureus virulence and physiology. PMID:25073642
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Strumila, Robertas; Lengvenytė, Aistė; Vainutienė, Vija; Lesinskas, Eugenijus
2017-12-01
Psychological factors have been described as important for tinnitus severity, but attempts to incorporate them in one picture are sparse. This study investigated to what extent traits (personality), states (depressive and anxiety symptoms), sociodemographic factors and questioning environment influence tinnitus severity perception and how they interplay. Data were obtained from 212 subjects in a survey that was undertaken in 2016 at Vilnius University hospital and via internet. Measures included the Tinnitus Handicap Inventory (THI), Visual Analogue Scale (VAS), Hospital Anxiety and Depression Scale (HADS), Big Five Personality Dimensions Scale and sociodemographic questions. A series of stepwise forward and multiple regression analyses were undertaken to discover how factors interconnect. Female gender, age, living in rural area, but not level of education, were found to be associated with THI and HADS. Total HADS score and of both subscales were linked to scores on THI, VAS scales and all personality traits, except agreeableness (and consciousness for anxiety). Anxiety was the most important predictor for tinnitus severity, followed by depressive symptoms. Only neuroticism from personality dimensions was a predictor of THI score, whereas THI scores did not predict scores on neuroticism. All results in scales were higher in the internet group, except agreeableness and neuroticism, while extroversion correlated negatively with THI score only in the hospital group. Tinnitus severity was highly correlated with depressive, anxiety symptoms and neuroticism. Respondents recruited through internet had higher scores on most parameters. Results emphasize the importance of psychological factors in tinnitus management.
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Analysis of lipid raft molecules in the living brain slices.
Kotani, Norihiro; Nakano, Takanari; Ida, Yui; Ito, Rina; Hashizume, Miki; Yamaguchi, Arisa; Seo, Makoto; Araki, Tomoyuki; Hojo, Yasushi; Honke, Koichi; Murakoshi, Takayuki
2017-08-24
Neuronal plasma membrane has been thought to retain a lot of lipid raft components which play important roles in the neural function. Although the biochemical analyses of lipid raft using brain tissues have been extensively carried out in the past 20 years, many of their experimental conditions do not coincide with those of standard neuroscience researches such as neurophysiology and neuropharmacology. Hence, the physiological methods for lipid raft analysis that can be compatible with general neuroscience have been required. Herein, we developed a system to physiologically analyze ganglioside GM1-enriched lipid rafts in brain tissues using the "Enzyme-Mediated Activation of Radical Sources (EMARS)" method that we reported (Kotani N. et al. Proc. Natl. Acad. Sci. U S A 105, 7405-7409 (2008)). The EMARS method was applied to acute brain slices prepared from mouse brains in aCSF solution using the EMARS probe, HRP-conjugated cholera toxin subunit B, which recognizes ganglioside GM1. The membrane molecules present in the GM1-enriched lipid rafts were then labeled with fluorescein under the physiological condition. The fluorescein-tagged lipid raft molecules called "EMARS products" distributed differentially among various parts of the brain. On the other hand, appreciable differences were not detected among segments along the longitudinal axis of the hippocampus. We further developed a device to label the lipid raft molecules in acute hippocampal slices under two different physiological conditions to detect dynamics of the lipid raft molecules during neural excitation. Using this device, several cell membrane molecules including Thy1, known as a lipid raft resident molecule in neurons, were confirmed by the EMARS method in living hippocampal slices. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Taylor, Hannah; Schmiedt, Joscha T.; Çarçak, Nihan; Onat, Filiz; Di Giovanni, Giuseppe; Lambert, Régis; Leresche, Nathalie; Crunelli, Vincenzo; David, Francois
2014-01-01
Background The advent of optogenetics has given neuroscientists the opportunity to excite or inhibit neuronal population activity with high temporal resolution and cellular selectivity. Thus, when combined with recordings of neuronal ensemble activity in freely moving animals optogenetics can provide an unprecedented snapshot of the contribution of neuronal assemblies to (patho)physiological conditions in vivo. Still, the combination of optogenetic and silicone probe (or tetrode) recordings does not allow investigation of the role played by voltage- and transmitter-gated channels of the opsin-transfected neurons and/or other adjacent neurons in controlling neuronal activity. New method and results We demonstrate that optogenetics and silicone probe recordings can be combined with intracerebral reverse microdialysis for the long-term delivery of neuroactive drugs around the optic fiber and silicone probe. In particular, we show the effect of antagonists of T-type Ca2+ channels, hyperpolarization-activated cyclic nucleotide-gated channels and metabotropic glutamate receptors on silicone probe-recorded activity of the local opsin-transfected neurons in the ventrobasal thalamus, and demonstrate the changes that the block of these thalamic channels/receptors brings about in the network dynamics of distant somatotopic cortical neuronal ensembles. Comparison with existing methods This is the first demonstration of successfully combining optogenetics and neuronal ensemble recordings with reverse microdialysis. This combination of techniques overcomes some of the disadvantages that are associated with the use of intracerebral injection of a drug-containing solution at the site of laser activation. Conclusions The combination of reverse microdialysis, silicone probe recordings and optogenetics can unravel the short and long-term effects of specific transmitter- and voltage-gated channels on laser-modulated firing at the site of optogenetic stimulation and the actions that these manipulations exert on distant neuronal populations. PMID:25004203
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Taylor, Hannah; Schmiedt, Joscha T; Carçak, Nihan; Onat, Filiz; Di Giovanni, Giuseppe; Lambert, Régis; Leresche, Nathalie; Crunelli, Vincenzo; David, Francois
2014-09-30
The advent of optogenetics has given neuroscientists the opportunity to excite or inhibit neuronal population activity with high temporal resolution and cellular selectivity. Thus, when combined with recordings of neuronal ensemble activity in freely moving animals optogenetics can provide an unprecedented snapshot of the contribution of neuronal assemblies to (patho)physiological conditions in vivo. Still, the combination of optogenetic and silicone probe (or tetrode) recordings does not allow investigation of the role played by voltage- and transmitter-gated channels of the opsin-transfected neurons and/or other adjacent neurons in controlling neuronal activity. We demonstrate that optogenetics and silicone probe recordings can be combined with intracerebral reverse microdialysis for the long-term delivery of neuroactive drugs around the optic fiber and silicone probe. In particular, we show the effect of antagonists of T-type Ca(2+) channels, hyperpolarization-activated cyclic nucleotide-gated channels and metabotropic glutamate receptors on silicone probe-recorded activity of the local opsin-transfected neurons in the ventrobasal thalamus, and demonstrate the changes that the block of these thalamic channels/receptors brings about in the network dynamics of distant somatotopic cortical neuronal ensembles. This is the first demonstration of successfully combining optogenetics and neuronal ensemble recordings with reverse microdialysis. This combination of techniques overcomes some of the disadvantages that are associated with the use of intracerebral injection of a drug-containing solution at the site of laser activation. The combination of reverse microdialysis, silicone probe recordings and optogenetics can unravel the short and long-term effects of specific transmitter- and voltage-gated channels on laser-modulated firing at the site of optogenetic stimulation and the actions that these manipulations exert on distant neuronal populations. Copyright © 2014. Published by Elsevier B.V.
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Gorniak, Tobias; Meyer, Ulrich; Südekum, Karl-Heinz; Dänicke, Sven
2014-01-01
The aim of the present study was to evaluate the impact of summer temperatures in a temperate climate on mid-lactation Holstein dairy cows. Therefore, a data set was examined comprising five trials with dairy cows conducted at the experimental station of the Friedrich-Loeffler-Institute in Braunschweig, Germany. The temperature-humidity index (THI) was calculated using temperature and humidity data from the barns recorded between January 2010 and July 2012. By using a generalised additive mixed model, the impact of increasing THI on dry matter intake, milk yield and milk composition was evaluated. Dry matter intake and milk yield decreased when THI rose above 60, whilst water intake increased in a linear manner beyond THI 30. Furthermore, milk protein and milk fat content decreased continuously with increasing THI. The present results revealed that heat stress exists in Lower Saxony, Germany. However, further research is necessary to describe the mode of action of heat stress. Especially, mild heat stress has to be investigated in more detail and appropriate heat stress thresholds for temperate climates have to be developed.
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Carcaud, Julie; Giurfa, Martin; Sandoz, Jean Christophe
2016-01-01
The function of parallel neural processing is a fundamental problem in Neuroscience, as it is found across sensory modalities and evolutionary lineages, from insects to humans. Recently, parallel processing has attracted increased attention in the olfactory domain, with the demonstration in both insects and mammals that different populations of second-order neurons encode and/or process odorant information differently. Among insects, Hymenoptera present a striking olfactory system with a clear neural dichotomy from the periphery to higher-order centers, based on two main tracts of second-order (projection) neurons: the medial and lateral antennal lobe tracts (m-ALT and l-ALT). To unravel the functional role of these two pathways, we combined specific lesions of the m-ALT tract with behavioral experiments, using the classical conditioning of the proboscis extension response (PER conditioning). Lesioned and intact bees had to learn to associate an odorant (1-nonanol) with sucrose. Then the bees were subjected to a generalization procedure with a range of odorants differing in terms of their carbon chain length or functional group. We show that m-ALT lesion strongly affects acquisition of an odor-sucrose association. However, lesioned bees that still learned the association showed a normal gradient of decreasing generalization responses to increasingly dissimilar odorants. Generalization responses could be predicted to some extent by in vivo calcium imaging recordings of l-ALT neurons. The m-ALT pathway therefore seems necessary for normal classical olfactory conditioning performance. PMID:26834589
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What Does the Thyroid Gland Do?
... it helps other cells do their job. hypothyroidism (hi-poh-THY-royd-izm): when your thyroid gland ... thyroid hormone (“hypo” means ‘under’ or ‘below’). hyperthyroidism (hi-purr-THY-royd-izm): when your thyroid gland ...
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Hsieh, Anne M-Y; Polyakova, Olena; Fu, Guodong; Chazen, Ronald S; MacMillan, Christina; Witterick, Ian J; Ralhan, Ranju; Walfish, Paul G
2018-04-13
Recognition of noninvasive follicular thyroid neoplasms with papillary-like nuclear features (NIFTP) that distinguishes them from invasive malignant encapsulated follicular variant of papillary thyroid carcinoma (EFVPTC) can prevent overtreatment of NIFTP patients. We and others have previously reported that programmed death-ligand 1 (PD-L1) is a useful biomarker in thyroid tumors; however, all reports to date have relied on manual scoring that is time consuming as well as subject to individual bias. Consequently, we developed a digital image analysis (DIA) protocol for cytoplasmic and membranous stain quantitation (ThyApp) and evaluated three tumor sampling methods [Systemic Uniform Random Sampling, hotspot nucleus, and hotspot nucleus/3,3'-Diaminobenzidine (DAB)]. A patient cohort of 153 cases consisting of 48 NIFTP, 44 EFVPTC, 26 benign nodules and 35 encapsulated follicular lesions/neoplasms with lymphocytic thyroiditis (LT) was studied. ThyApp quantitation of PD-L1 expression revealed a significant difference between invasive EFVPTC and NIFTP; but none between NIFTP and benign nodules. ThyApp integrated with hotspot nucleus tumor sampling method demonstrated to be most clinically relevant, consumed least processing time, and eliminated interobserver variance. In conclusion, the fully automatic DIA algorithm developed using a histomorphological approach objectively quantitated PD-L1 expression in encapsulated thyroid neoplasms and outperformed manual scoring in reproducibility and higher efficiency.
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Veraitch, Ophelia; Mabuchi, Yo; Matsuzaki, Yumi; Sasaki, Takashi; Okuno, Hironobu; Tsukashima, Aki; Amagai, Masayuki; Okano, Hideyuki; Ohyama, Manabu
2017-01-01
The dermal papilla (DP) is a specialised mesenchymal component of the hair follicle (HF) that plays key roles in HF morphogenesis and regeneration. Current technical difficulties in preparing trichogenic human DP cells could be overcome by the use of highly proliferative and plastic human induced pluripotent stem cells (hiPSCs). In this study, hiPSCs were differentiated into induced mesenchymal cells (iMCs) with a bone marrow stromal cell phenotype. A highly proliferative and plastic LNGFR(+)THY-1(+) subset of iMCs was subsequently programmed using retinoic acid and DP cell activating culture medium to acquire DP properties. The resultant cells (induced DP-substituting cells [iDPSCs]) exhibited up-regulated DP markers, interacted with human keratinocytes to up-regulate HF related genes, and when co-grafted with human keratinocytes in vivo gave rise to fibre structures with a hair cuticle-like coat resembling the hair shaft, as confirmed by scanning electron microscope analysis. Furthermore, iDPSCs responded to the clinically used hair growth reagent, minoxidil sulfate, to up-regulate DP genes, further supporting that they were capable of, at least in part, reproducing DP properties. Thus, LNGFR(+)THY-1(+) iMCs may provide material for HF bioengineering and drug screening for hair diseases. PMID:28220862
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The rational use of fine needle aspiration biopsy (FNAB) in diagnosing thyroid nodules.
Sidoti, M; Marino, G; Resmini, E; Augeri, C; Cappi, C; Cavallero, D; Lagasio, C; Ceppa, P; Minuto, F; Giusti, M
2006-06-01
Fine needle aspiration biopsy (FNAB) plays a crucial role in the diagnosis of thyroid nodules and enables the number of surgical operations to be reduced. Theoretically, FNAB should be carried out on all nodules, though currently only those displaying certain characteristics are biopsied. Indeed, to perform FNAB on all nodules may be regarded as an excess of zeal. Therefore, it seems advisable that the endocrinologist should be able to confirm on the spot the necessity and utility of FNAB. We evaluated on a sample of 263 consecutive requests (209 female, 57 male; age 56.7+/-13.7 years) for FNAB in 2004: 1) the appropriateness of the investigation, 2) expected efficacy, 3) practical efficacy, 4) efficiency. FNAB was performed under echo-guidance in accordance with the standard technique. In 50%, 36%, 6%, 3%, 2% and 1% of cases, the echographic diagnosis was of MNG, UNG, pseudo-nodular lesion in ATD, lymph-node, neck cyst, suspected parathyroid lesion and tumefaction of the salivary glands, respectively. A pre-FNAB clinical risk score was assigned to each case on the basis of clinical and echographic data, with a maximum possible score of 11. The results of FNAB were subdivided into 5 categories according to the criteria of the BTA (Thy1-Thy5). After FNAB, a decisional category was assigned, ranging from ''observation'' to ''surgery''; this was subsequently (7-18 months) compared with the management strategy adopted by the attending physician. Information was gathered by means of telephone enquiry. 1) Appropriateness: on the basis of clinical and echographic findings, FNAB was not judged appropriate in 24% of cases because of either the lack of confirmation of a significant target (34%) or a low pre-FNAB risk score (range 0-2) (66%). The decisional category was ''observation'' in 87% of cases and ''further investigation'' in 13%. 2) Expected efficacy: FNAB was performed in 76% of cases. The biopsies (3%) performed on swollen lymph-nodes and extra-thyroid neck tumefactions, in which biochemical evaluation was positive, proved to be diagnostic but not classifiable according to the BTA. In 82% of the remaining cases, the result was Thy2 (observation) or Thy 4-5 (surgery). Thy3 results (surgery) were rare (1%). Thy1 results (16%) were yielded by the aspiration of colloid cysts (29%), solid lesions (10%) characterised by means of PTH-FNAB and Tg-FNAB, nodules (9%) no longer detectable on repetition of FNAB, nodules (16%) in which FNAB was already a repetition of a non-diagnostic investigation (2003), and nodules (9%) in which the presence of normal thyrocytes, ''hot'' scintigraphic image and prior decision of the surgeon advised against repeating FNAB. Of the patients with Thy1 results, 26% refused to repeat FNAB. In all, 95% of FNAB supported by biochemical evaluation yielded results that usefully contributed to patient management. The correlation between pre-FNAB clinical risk and cytological score according to the BTA proved significant (P<0.001). No difference in diameter was recorded between nodules with adequate cytology (23.3+/-0.9 mm) and those with inadequate cytology (25.2+/-1.6 mm). 3) Practical efficacy: 75% of patients were reached by telephone. In most cases, observation was the most frequent clinical choice, after echography and/or FNAB. The decisional category assigned after FNAB correlated significantly (P<0.001) with the approach adopted by the attending physician. d) Efficiency: following FNAB, 11 patients were assigned to surgery. DTC was detected in 100% of these cases (1 follicular carcinoma, 1 insular carcinoma, 9 papillary carcinoma). The success of FNAB (9/11) in detecting lesions that proved malignant on histological examination (11/11) was significant (P<0.05). Of the 2 Thy 3 cases, 1 was follicular carcinoma and 1 was follicular adenoma with adjacent papillary carcinoma. The incidence of thyroid carcinomas in the population studied was 5.5%. 1) Together with clinical-biochemical evaluation, echo-guided FNAB re-mains the first-line diagnostic test in the management of thyroid nodules; 2) a pre-FNAB clinical risk score is useful in limiting the number of probably inappropriate investigations; 3) efficacy, in terms of cytology results that are useful for patient management after FNAB (and after biochemical evaluation, when indicated) is high, enabling patients to be stratified in classes with different subsequent pathways; 4) in the vast majority of cases, FNAB influences subsequent clinical decisions; 5) false negatives cannot be excluded, while false positives are practically nil; 6) further indications may be yielded by studies on larger populations, and new prospects may emerge from the application of other techniques associated to FNAB.
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Wrzosek, Małgorzata; Szymiec, Eugeniusz; Klemens, Wiesława; Kotyło, Piotr; Schlee, Winfried; Modrzyńska, Małgorzata; Lang-Małecka, Agnieszka; Preis, Anna; Bulla, Jan
2016-01-01
Objective: The need for validated measures enabling clinicians to classify tinnitus patients according to the severity of tinnitus and screen the progress of therapies in our country led us to translate into Polish and to validate two tinnitus questionnaires, namely the Tinnitus Handicap Inventory (THI) and the Tinnitus Functional Index (TFI). Design: The original English versions of the questionnaires were translated into Polish and translated back to English by three independent translators. These versions were then finalized by the authors into a Polish THI (THI-Pl) and a Polish TFI (TFI-Pl). Participants from three laryngological centers in Poland anonymously answered the THI-Pl (N = 98) and the TFI-Pl (N = 108) in addition to the Polish versions of the Center for Epidemiologic Studies Depression Scale as a measure of self-perceived level of depression, and the Satisfaction With Life Scale to assess self-perceived quality of life. Both were used to determine discriminant validity. Two Visual Analog Scales were used to measure tinnitus annoyance and tinnitus loudness in order to determine convergent validity. Results: Similar to the original version of the THI, the THI-Pl showed a high internal consistency (Cronbach’s α = 0.93). The exploratory factor analysis revealed that the questionnaire has a three-factorial structure that does not correspond to the original division for functional, catastrophic, and emotional subscales. Convergent and discriminant validities were confirmed. The TFI-Pl showed high internal consistency (Cronbach’s α = 0.96) with the reliability ranging from 0.82 to 0.95 for its different subscales. Factor analysis confirmed an eight-factorial structure with factors assigning all items to appropriate subscales reported in the original version of the questionnaire. Discriminant and convergent validities were also confirmed for the TFI-Pl. Conclusion: We translated and validated the Polish versions of the THI and the TFI to make them suitable for clinical use in Poland. PMID:27965609
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Myeloid Conditioning with c-kit-Targeted CAR-T Cells Enables Donor Stem Cell Engraftment.
Arai, Yasuyuki; Choi, Uimook; Corsino, Cristina I; Koontz, Sherry M; Tajima, Masaki; Sweeney, Colin L; Black, Mary A; Feldman, Steven A; Dinauer, Mary C; Malech, Harry L
2018-05-02
We report a novel approach to bone marrow (BM) conditioning using c-kit-targeted chimeric antigen receptor T (c-kit CAR-T) cells in mice. Previous reports using anti-c-kit or anti-CD45 antibody linked to a toxin such as saporin have been promising. We developed a distinctly different approach using c-kit CAR-T cells. Initial studies demonstrated in vitro killing of hematopoietic stem cells by c-kit CAR-T cells but poor expansion in vivo and poor migration of CAR-T cells into BM. Pre-treatment of recipient mice with low-dose cyclophosphamide (125 mg/kg) together with CXCR4 transduction in the CAR-T cells enhanced trafficking to and expansion in BM (<1%-13.1%). This resulted in significant depletion of the BM c-kit + population (9.0%-0.1%). Because congenic Thy1.1 CAR-T cells were used in the Thy1.2-recipient mice, anti-Thy1.1 antibody could be used to deplete CAR-T cells in vivo before donor BM transplant. This achieved 20%-40% multilineage engraftment. We applied this conditioning to achieve an average of 28% correction of chronic granulomatous disease mice by wild-type BM transplant. Our findings provide a proof of concept that c-kit CAR-T cells can achieve effective BM conditioning without chemo-/radiotherapy. Our work also demonstrates that co-expression of a trafficking receptor can enhance targeting of CAR-T cells to a designated tissue. Published by Elsevier Inc.
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Munemasa, Yasunari; Chang, Chang-Sheng; Kwong, Jacky M K; Kyung, Haksu; Kitaoka, Yasushi; Caprioli, Joseph; Piri, Natik
2012-01-01
Nell2 is a neuron-specific protein containing six epidermal growth factor-like domains. We have identified Nell2 as a retinal ganglion cell (RGC)-expressed gene by comparing mRNA profiles of control and RGC-deficient rat retinas. The aim of this study was to analyze Nell2 expression in wild-type and optic nerve axotomized retinas and evaluate its potential role in RGCs. Nell2-positive in situ and immunohistochemical signals were localized to irregularly shaped cells in the ganglion cell layer (GCL) and colocalized with retrogradely-labeled RGCs. No Nell2-positive cells were detected in 2 weeks optic nerve transected (ONT) retinas characterized with approximately 90% RGC loss. RT-PCR analysis showed a dramatic decrease in the Nell2 mRNA level after ONT compared to the controls. Immunoblot analysis of the Nell2 expression in the retina revealed the presence of two proteins with approximate MW of 140 and 90 kDa representing glycosylated and non-glycosylated Nell2, respectively. Both products were almost undetectable in retinal protein extracts two weeks after ONT. Proteome analysis of Nell2-interacting proteins carried out with MALDI-TOF MS (MS) identified microtubule-actin crosslinking factor 1 (Macf1), known to be critical in CNS development. Strong Macf1 expression was observed in the inner plexiform layer and GCL where it was colocalizied with Thy-1 staining. Since Nell2 has been reported to increase neuronal survival of the hippocampus and cerebral cortex, we evaluated the effect of Nell2 overexpression on RGC survival. RGCs in the nasal retina were consistently more efficiently transfected than in other areas (49% vs. 13%; n = 5, p<0.05). In non-transfected or pEGFP-transfected ONT retinas, the loss of RGCs was approximately 90% compared to the untreated control. In the nasal region, Nell2 transfection led to the preservation of approximately 58% more cells damaged by axotomy compared to non-transfected (n = 5, p<0.01) or pEGFP-transfected controls (n = 5, p<0.01).
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Munemasa, Yasunari; Chang, Chang-Sheng; Kwong, Jacky M. K.; Kyung, Haksu; Kitaoka, Yasushi; Caprioli, Joseph; Piri, Natik
2012-01-01
Nell2 is a neuron-specific protein containing six epidermal growth factor-like domains. We have identified Nell2 as a retinal ganglion cell (RGC)-expressed gene by comparing mRNA profiles of control and RGC-deficient rat retinas. The aim of this study was to analyze Nell2 expression in wild-type and optic nerve axotomized retinas and evaluate its potential role in RGCs. Nell2-positive in situ and immunohistochemical signals were localized to irregularly shaped cells in the ganglion cell layer (GCL) and colocalized with retrogradely-labeled RGCs. No Nell2-positive cells were detected in 2 weeks optic nerve transected (ONT) retinas characterized with approximately 90% RGC loss. RT-PCR analysis showed a dramatic decrease in the Nell2 mRNA level after ONT compared to the controls. Immunoblot analysis of the Nell2 expression in the retina revealed the presence of two proteins with approximate MW of 140 and 90 kDa representing glycosylated and non-glycosylated Nell2, respectively. Both products were almost undetectable in retinal protein extracts two weeks after ONT. Proteome analysis of Nell2-interacting proteins carried out with MALDI-TOF MS (MS) identified microtubule-actin crosslinking factor 1 (Macf1), known to be critical in CNS development. Strong Macf1 expression was observed in the inner plexiform layer and GCL where it was colocalizied with Thy-1 staining. Since Nell2 has been reported to increase neuronal survival of the hippocampus and cerebral cortex, we evaluated the effect of Nell2 overexpression on RGC survival. RGCs in the nasal retina were consistently more efficiently transfected than in other areas (49% vs. 13%; n = 5, p<0.05). In non-transfected or pEGFP-transfected ONT retinas, the loss of RGCs was approximately 90% compared to the untreated control. In the nasal region, Nell2 transfection led to the preservation of approximately 58% more cells damaged by axotomy compared to non-transfected (n = 5, p<0.01) or pEGFP-transfected controls (n = 5, p<0.01). PMID:22496866
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García-Berrocoso, Teresa; Llombart, Víctor; Colàs-Campàs, Laura; Hainard, Alexandre; Licker, Virginie; Penalba, Anna; Ramiro, Laura; Simats, Alba; Bustamante, Alejandro; Martínez-Saez, Elena; Canals, Francesc; Sanchez, Jean-Charles; Montaner, Joan
2018-01-01
Cerebral ischemia entails rapid tissue damage in the affected brain area causing devastating neurological dysfunction. How each component of the neurovascular unit contributes or responds to the ischemic insult in the context of the human brain has not been solved yet. Thus, the analysis of the proteome is a straightforward approach to unraveling these cell proteotypes. In this study, post-mortem brain slices from ischemic stroke patients were obtained corresponding to infarcted (IC) and contralateral (CL) areas. By means of laser microdissection, neurons and blood brain barrier structures (BBB) were isolated and analyzed using label-free quantification. MS data are available via ProteomeXchange with identifier PXD003519. Ninety proteins were identified only in neurons, 260 proteins only in the BBB and 261 proteins in both cell types. Bioinformatics analyses revealed that repair processes, mainly related to synaptic plasticity, are outlined in microdissected neurons, with nonexclusive important functions found in the BBB. A total of 30 proteins showing p < 0.05 and fold-change> 2 between IC and CL areas were considered meaningful in this study: 13 in neurons, 14 in the BBB and 3 in both cell types. Twelve of these proteins were selected as candidates and analyzed by immunohistofluorescence in independent brains. The MS findings were completely verified for neuronal SAHH2 and SRSF1 whereas the presence in both cell types of GABT and EAA2 was only validated in neurons. In addition, SAHH2 showed its potential as a prognostic biomarker of neurological improvement when analyzed early in the plasma of ischemic stroke patients. Therefore, the quantitative proteomes of neurons and the BBB (or proteotypes) after human brain ischemia presented here contribute to increasing the knowledge regarding the molecular mechanisms of ischemic stroke pathology and highlight new proteins that might represent putative biomarkers of brain ischemia or therapeutic targets. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.
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Identification of HIV-1 determinants for replication in vivo.
Su, L; Kaneshima, H; Bonyhadi, M L; Lee, R; Auten, J; Wolf, A; Du, B; Rabin, L; Hahn, B H; Terwilliger, E; Mccune, J M
1997-01-06
Pathogenic organisms are frequently attenuated after long-term culture in vitro. The mechanisms of the attenuation process are not clear, but probably involve mutations of functions required for replication and pathogenicity in vivo. To identify these functions, a direct comparison must be made between attenuated genomes and those that remain pathogenic in vivo. In this study, we used the heterochimeric SCID-hu Thy/Liv mouse as an in vivo model to define human immunodeficiency virus type 1 (HIV-1) determinants which are uniquely required for replication in vivo. The Lai/IIIB isolate and its associated infectious molecular clones (e.g., HXB2) were found to infect T cell lines but failed to replicate in the SCID-hu Thy/Liv model. When a lab worker was accidentally infected by Lai/IIIB, however, HIV-1 was isolated only from infection of primary PBMC, and not from infection of T cell lines. We hypothesized that the lab worker was exposed to a heterogeneous viral stock which had been attenuated by passage in immortalized T cell lines. Either a rare family member from this stock was selected for in vivo replication or, alternatively, an attenuated genotype dominant in vitro may have reverted to become more infectious in vivo. To address this hypothesis, we have used the SCID-hu Thy/Liv model to study the replication of HXB2 and of HXB2 recombinant viruses with HIV-1 fragments isolated from the infected lab worker. HXB2 showed no or very low levels of replication in the Thy/Liv organ. Replacement of its subgenomic fragment encoding the envelope gene with a corresponding fragment from the lab worker isolate generated a recombinant virus (HXB2/LW) which replicated actively in SCID-hu mice. The NEF mutation in the HXB2 genome is still present in HXB2/LW. Thus, the LW sequences encode HIV-1 determinants which enhance HIV replication in vivo in a NEF-independent mechanism. The specific determinants have been mapped to the V1-V3 regions of the HIV-1 genome. Six unique mutations in the V3 loop region of HXB2/LW have been identified which contribute to the increased replication in vivo.
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Reliability and validity of the Lithuanian Tinnitus Handicap Inventory.
Ulozienė, Ingrida; Balnytė, Renata; Alzbutienė, Giedrė; Arechvo, Irina; Vaitkus, Antanas; Šileikaitė, Milda; Šaferis, Viktoras; Ulozas, Virgilijus
2016-01-01
The aim of this study was to determine the reliability and validity of the Lithuanian version of the Tinnitus Handicap Inventory (THI), a self-report measure of perceived tinnitus handicap. A cross-sectional psychometric validation study was performed in the University Hospital. A total of 248 subjects reporting chronic tinnitus as their primary complaint or secondary to hearing loss were encluded in the study and filled in the Lithuanian version of THI. For assessment of construct validity a subgroup of 55 participants completed the Lithuanian version of the Hospital Anxiety and Depression Scale as a measure of self-perceived levels of anxiety and depression. Test-retest and internal consistency reliability as well as construct validity were calculated. The Lithuanian version of the THI and its subscales showed a robust internal consistency reliability (Cronbach's alpha=0.93) comparable to the original version. Statistically significant correlations were observed between the Lithuanian translation of the THI and the measures of self-perceived levels of anxiety and depression using HADS. Confirmatory factor analysis demonstrated that the three subscales of the THI Lithuanian version corresponded to three different factors, which strongly correlated between themselves. The results suggest that the Lithuanian version of THI maintains its original validity and may serve as reliable and valid measure of general tinnitus related distress that can be used in a clinical setting to quantify the impact of tinnitus on daily living. Copyright © 2016 The Lithuanian University of Health Sciences. Production and hosting by Elsevier Urban & Partner Sp. z o.o. All rights reserved.
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Challand, Martin R.; Martins, Filipa T.; Roach, Peter L.
2010-01-01
Thiazole synthase in Escherichia coli is an αβ heterodimer of ThiG and ThiH. ThiH is a tyrosine lyase that cleaves the Cα–Cβ bond of tyrosine, generating p-cresol as a by-product, to form dehydroglycine. This reactive intermediate acts as one of three substrates for the thiazole cyclization reaction catalyzed by ThiG. ThiH is a radical S-adenosylmethionine (AdoMet) enzyme that utilizes a [4Fe-4S]+ cluster to reductively cleave AdoMet, forming methionine and a 5′-deoxyadenosyl radical. Analysis of the time-dependent formation of the reaction products 5′-deoxyadenosine (DOA) and p-cresol has demonstrated catalytic behavior of the tyrosine lyase. The kinetics of product formation showed a pre-steady state burst phase, and the involvement of DOA in product inhibition was identified by the addition of 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidase to activity assays. This hydrolyzed the DOA and changed the rate-determining step but, in addition, substantially increased the uncoupled turnover of AdoMet. Addition of glyoxylate and ammonium inhibited the tyrosine cleavage reaction, but the reductive cleavage of AdoMet continued in an uncoupled manner. Tyrosine analogues were incubated with ThiGH, which showed a strong preference for phenolic substrates. 4-Hydroxyphenylpropionic acid analogues allowed uncoupled AdoMet cleavage but did not result in further reaction (Cα–Cβ bond cleavage). The results of the substrate analogue studies and the product inhibition can be explained by a mechanistic hypothesis involving two reaction pathways, a product-forming pathway and a futile cycle. PMID:19923213
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Dual Anterograde and Retrograde Viral Tracing of Reciprocal Connectivity.
Haberl, Matthias G; Ginger, Melanie; Frick, Andreas
2017-01-01
Current large-scale approaches in neuroscience aim to unravel the complete connectivity map of specific neuronal circuits, or even the entire brain. This emerging research discipline has been termed connectomics. Recombinant glycoprotein-deleted rabies virus (RABV ∆G) has become an important tool for the investigation of neuronal connectivity in the brains of a variety of species. Neuronal infection with even a single RABV ∆G particle results in high-level transgene expression, revealing the fine-detailed morphology of all neuronal features-including dendritic spines, axonal processes, and boutons-on a brain-wide scale. This labeling is eminently suitable for subsequent post-hoc morphological analysis, such as semiautomated reconstruction in 3D. Here we describe the use of a recently developed anterograde RABV ∆G variant together with a retrograde RABV ∆G for the investigation of projections both to, and from, a particular brain region. In addition to the automated reconstruction of a dendritic tree, we also give as an example the volume measurements of axonal boutons following RABV ∆G-mediated fluorescent marker expression. In conclusion RABV ∆G variants expressing a combination of markers and/or tools for stimulating/monitoring neuronal activity, used together with genetic or behavioral animal models, promise important insights in the structure-function relationship of neural circuits.
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NASA Astrophysics Data System (ADS)
Bernabucci, U.; Lacetera, N.; Danieli, P. P.; Bani, P.; Nardone, A.; Ronchi, B.
2009-09-01
Effects of different periods of exposure to hot environments on rumen function, diet digestibility and digesta passage rate were studied in four adult not-pregnant Sardinian ewes housed in a climatic chamber. The ewes were kept in individual metabolic cages. The trial lasted 83 days; 17 days were spent under thermal comfort conditions (TC) [temperature-humidity index (THI) = 65.0 ± 2.0], followed by 49 days under elevated THI (ETHI: THI = 82.0 ± 2.5) and 17 days under thermal comfort (TC; THI = 65.0 ± 1.0). Five digestibility and passage rate trials were carried out during the 83 days. Trials 1 and 5 were carried out under TC; trials 2, 3 and 4 were carried out under ETHI. Values of rectal temperatures (39.7 ± 0.3°C) and respiratory rate (118.4 ± 31.8 breaths/min) indicated that sheep under ETHI were heat-stressed. Heat stress caused an increase ( P < 0.01) in water intake, and reductions ( P < 0.05) in dry matter intake, rumen pH, rumen cellulolytic and amylolytic bacteria count, rumen osmolarity, organic matter, dry matter, neutral detergent fibre, acid detergent fibre and non-structural carbohydrates digestibility coefficients, and a reduction of digesta passage rates. Under ETHI, diet digestibility and passage rate of digesta were reduced in a time-dependent fashion. Variation of diet digestibility under ETHI was not related to passage rate of digesta and feed intake. Reduction of cellulolytic and amylolytic bacteria and the adaptive response to hot environment seem to be related to alteration of digestibility observed in ewes chronically exposed to hot environment.
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Simplified form of tinnitus retraining therapy in adults: a retrospective study
Aazh, Hashir; Moore, Brian CJ; Glasberg, Brian R
2008-01-01
Background Since the first description of tinnitus retraining therapy (TRT), clinicians have modified and customised the method of TRT in order to suit their practice and their patients. A simplified form of TRT is used at Ealing Primary Care Trust Audiology Department. Simplified TRT is different from TRT in the type and (shorter) duration of the counseling but is similar to TRT in the application of sound therapy except for patients exhibiting tinnitus with no hearing loss and no decreased sound tolerance (wearable sound generators were not mandatory or recommended here, whereas they are for TRT). The main goal of this retrospective study was to assess the efficacy of simplified TRT. Methods Data were collected from a series of 42 consecutive patients who underwent simplified TRT for a period of 3 to 23 months. Perceived tinnitus handicap was measured by the Tinnitus Handicap Inventory (THI) and perceived tinnitus loudness, annoyance and the effect of tinnitus on life were assessed through the Visual Analog Scale (VAS). Results The mean THI and VAS scores were significantly decreased after 3 to 23 months of treatment. The mean decline of the THI score was 45 (SD = 22) and the difference between pre- and post-treatment scores was statistically significant. The mean decline of the VAS scores was 1.6 (SD = 2.1) for tinnitus loudness, 3.6 (SD = 2.6) for annoyance, and 3.9 (SD = 2.3) for effect on life. The differences between pre- and post-treatment VAS scores were statistically significant for tinnitus loudness, annoyance, and effect on life. The decline of THI scores was not significantly correlated with age and duration of tinnitus. Conclusion The results suggest that benefit may be obtained from a substantially simplified form of TRT. PMID:18980672
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Hino, Kimihiro; Lee, Jung Su; Asami, Yasushi
2017-12-01
People's year-round interpersonal step count variations according to meteorological conditions are not fully understood, because complete year-round data from a sufficient sample of the general population are difficult to acquire. This study examined the associations between meteorological conditions and objectively measured step counts using year-round data collected from a large cohort ( N = 24,625) in Yokohama, Japan from April 2015 to March 2016. Two-piece linear regression analysis was used to examine the associations between the monthly median daily step count and three meteorological indices (mean values of temperature, temperature-humidity index (THI), and net effective temperature (NET)). The number of steps per day peaked at temperatures between 19.4 and 20.7 °C. At lower temperatures, the increase in steps per day was between 46.4 and 52.5 steps per 1 °C increase. At temperatures higher than those at which step counts peaked, the decrease in steps per day was between 98.0 and 187.9 per 1 °C increase. Furthermore, these effects were more obvious in elderly than non-elderly persons in both sexes. A similar tendency was seen when using THI and NET instead of temperature. Among the three meteorological indices, the highest R 2 value with step counts was observed with THI in all four groups. Both high and low meteorological indices discourage people from walking and higher values of the indices adversely affect step count more than lower values, particularly among the elderly. Among the three indices assessed, THI best explains the seasonal fluctuations in step counts.
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Simplified form of tinnitus retraining therapy in adults: a retrospective study.
Aazh, Hashir; Moore, Brian C J; Glasberg, Brian R
2008-11-03
Since the first description of tinnitus retraining therapy (TRT), clinicians have modified and customised the method of TRT in order to suit their practice and their patients. A simplified form of TRT is used at Ealing Primary Care Trust Audiology Department. Simplified TRT is different from TRT in the type and (shorter) duration of the counseling but is similar to TRT in the application of sound therapy except for patients exhibiting tinnitus with no hearing loss and no decreased sound tolerance (wearable sound generators were not mandatory or recommended here, whereas they are for TRT). The main goal of this retrospective study was to assess the efficacy of simplified TRT. Data were collected from a series of 42 consecutive patients who underwent simplified TRT for a period of 3 to 23 months. Perceived tinnitus handicap was measured by the Tinnitus Handicap Inventory (THI) and perceived tinnitus loudness, annoyance and the effect of tinnitus on life were assessed through the Visual Analog Scale (VAS). The mean THI and VAS scores were significantly decreased after 3 to 23 months of treatment. The mean decline of the THI score was 45 (SD = 22) and the difference between pre- and post-treatment scores was statistically significant. The mean decline of the VAS scores was 1.6 (SD = 2.1) for tinnitus loudness, 3.6 (SD = 2.6) for annoyance, and 3.9 (SD = 2.3) for effect on life. The differences between pre- and post-treatment VAS scores were statistically significant for tinnitus loudness, annoyance, and effect on life. The decline of THI scores was not significantly correlated with age and duration of tinnitus. The results suggest that benefit may be obtained from a substantially simplified form of TRT.
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NASA Astrophysics Data System (ADS)
Shivalingaiah, Shivaranjani; Gu, Ling; Mohanty, Samarendra K.
2011-03-01
Stimulation of retinal neuronal cells using optogenetics via use of chanelrhodopsin-2 (ChR2) and blue light has opened up a new direction for restoration of vision with respect to treatment of Retinitis pigmentosa (RP). In addition to delivery of ChR2 to specific retinal layer using genetic engineering, threshold level of blue light needs to be delivered onto the retina for generating action potential and successful behavioral outcome. We report measurement of intensity distribution of light reaching the retina of Retinitis pigmentosa (RP) mouse models and compared those results with theoretical simulations of light propagation in eye. The parameters for the stimulating source positioning in front of eye was determined for optimal light delivery to the retina. In contrast to earlier viral method based delivery of ChR2 onto retinal ganglion cells, in-vivo electroporation method was employed for retina-transfection of RP mice. The behavioral improvement in mice with Thy1-ChR2-YFP transfected retina, expressing ChR2 in retinal ganglion cells, was found to correlate with stimulation intensity.
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NASA Astrophysics Data System (ADS)
Davison, T. M.; Jonsson, N. N.; Mayer, D. G.; Gaughan, J. B.; Ehrlich, W. K.; McGowan, M. R.
2016-12-01
Exposure to hot environments affects milk yield (MY) and milk composition of pasture and feed-pad fed dairy cows in subtropical regions. This study was undertaken during summer to compare MY and physiology of cows exposed to six heat-load management treatments. Seventy-eight Holstein-Friesian cows were blocked by season of calving, parity, milk yield, BW, and milk protein (%) and milk fat (%) measured in 2 weeks prior to the start of the study. Within blocks, cows were randomly allocated to one of the following treatments: open-sided iron roofed day pen adjacent to dairy (CID) + sprinklers (SP); CID only; non-shaded pen adjacent to dairy + SP (NSD + SP); open-sided shade cloth roofed day pen adjacent to dairy (SCD); NSD + sprinkler (sprinkler on for 45 min at 1100 h if mean respiration rate >80 breaths per minute (NSD + WSP) ); open-sided shade cloth roofed structure over feed bunk in paddock + 1 km walk to and from the dairy (SCP + WLK). Sprinklers for CID + SP and NSD + SP cycled 2 min on, 12 min off when ambient temperature >26°C. The highest milk yields were in the CID + SP and CID treatments (23.9 L cow-1 day-1), intermediate for NSD + SP, SCD and SCP + WLK (22.4 L cow-1 day-1), and lowest for NSD + WSP (21.3 L cow-1 day-1) ( P < 0.05). The highest ( P < 0.05) feed intakes occurred in the CID + SP and CID treatments while intake was lowest ( P < 0.05) for NSD + WSP and SCP + WLK. Weather data were collected on site at 10-min intervals, and from these, THI was calculated. Nonlinear regression modelling of MY × THI and heat-load management treatment demonstrated that cows in CID + SP showed no decline in MY out to a THI break point value of 83.2, whereas the pooled MY of the other treatments declined when THI >80.7. A combination of iron roof shade plus water sprinkling throughout the day provided the most effective control of heat load.
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Cowley, F C; Barber, D G; Houlihan, A V; Poppi, D P
2015-04-01
The effects of heat stress on dairy production can be separated into 2 distinct causes: those effects that are mediated by the reduced voluntary feed intake associated with heat stress, and the direct physiological and metabolic effects of heat stress. To distinguish between these, and identify their effect on milk protein and casein concentration, mid-lactation Holstein-Friesian cows (n = 24) were housed in temperature-controlled chambers and either subjected to heat stress [HS; temperature-humidity index (THI) ~78] or kept in a THI<70 environment and pair-fed with heat-stressed cows (TN-R) for 7 d. A control group of cows was kept in a THI<70 environment with ad libitum feeding (TN-AL). A subsequent recovery period (7 d), with THI<70 and ad libitum feeding followed. Intake accounted for only part of the effects of heat stress. Heat stress reduced the milk protein concentration, casein number, and casein concentration and increased the urea concentration in milk beyond the effects of restriction of intake. Under HS, the proportion in total casein of αS1-casein increased and the proportion of αS2-casein decreased. Because no effect of HS on milk fat or lactose concentration was found, these effects appeared to be the result of specific downregulation of mammary protein synthesis, and not a general reduction in mammary activity. No residual effects were found of HS or TN-R on milk production or composition after THI<70 and ad libitum intake were restored. Heat-stressed cows had elevated blood concentrations of urea and Ca, compared with TN-R and TN-AL. Cows in TN-R had higher serum nonesterified fatty acid concentrations than cows in HS. It was proposed that HS and TN-R cows may mobilize different tissues as endogenous sources of energy. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
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Watt, Torquil; Barbesino, Giuseppe; Bjorner, Jakob Bue; Bonnema, Steen Joop; Bukvic, Branka; Drummond, Russell; Groenvold, Mogens; Hegedüs, Laszlo; Kantzer, Valeska; Lasch, Kathryn E; Marcocci, Claudio; Mishra, Anjali; Netea-Maier, Romana; Ekker, Merel; Paunovic, Ivan; Quinn, Terence J; Rasmussen, Åse Krogh; Russell, Audrey; Sabaretnam, Mayilvaganan; Smit, Johannes; Törring, Ove; Zivaljevic, Vladan; Feldt-Rasmussen, Ulla
2015-03-01
Thyroid diseases are common and often affect quality of life (QoL). No cross-culturally validated patient-reported outcome measuring thyroid-related QoL is available. The purpose of the present study was to test the cross-cultural validity of the newly developed thyroid-related patient-reported outcome ThyPRO, using tests for differential item functioning (DIF) according to language version. The ThyPRO consists of 85 items summarized in 13 multi-item scales and one single item. Scales cover physical and mental symptoms, well-being and function as well as social and daily function and cosmetic concerns. Translation applied standard forward-backward methodology with subsequent cognitive interviews and reviews. Responses (N = 1,810) to the ThyPRO were collected in seven countries: UK (n = 166), The Netherlands (n = 147), Serbia (n = 150), Italy (n = 110), India (n = 148), Denmark (n = 902) and Sweden (n = 187). Translated versions were compared pairwise to the English version by examining uniform and nonuniform DIF, i.e., whether patients from different countries respond differently to a particular item, although they have identical level of the concept measured by the item. Analyses were controlled for thyroid diagnosis. DIF was investigated by ordinal logistic regression, testing for both statistical significance and magnitude (ΔR (2) > 0.02). Scale level was estimated by the sum score, after purification. For twelve of the 84 tested items, DIF was identified in more than one language. Eight of these were small, but four were indicative of possible low translatability. Twenty-one instances of DIF in single languages were identified, indicating potential problems with the particular translation. However, only seven were of a magnitude which could affect scale scores, most of which could be explained by sample differences not controlled for. The ThyPRO has good cross-cultural validity with only minor cross-cultural invariance and is recommended for use in international multicenter studies.
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Belarbi, Karim; Burnouf, Sylvie; Fernandez-Gomez, Francisco-Jose; Laurent, Cyril; Lestavel, Sophie; Figeac, Martin; Sultan, Audrey; Troquier, Laetitia; Leboucher, Antoine; Caillierez, Raphaëlle; Grosjean, Marie-Eve; Demeyer, Dominique; Obriot, Hélène; Brion, Ingrid; Barbot, Bérangère; Galas, Marie-Christine; Staels, Bart; Humez, Sandrine; Sergeant, Nicolas; Schraen-Maschke, Susanna; Muhr-Tailleux, Anne; Hamdane, Malika; Buée, Luc; Blum, David
2011-08-01
Tau pathology is encountered in many neurodegenerative disorders known as tauopathies, including Alzheimer's disease. Physical activity is a lifestyle factor affecting processes crucial for memory and synaptic plasticity. Whether long-term voluntary exercise has an impact on Tau pathology and its pathophysiological consequences is currently unknown. To address this question, we investigated the effects of long-term voluntary exercise in the THY-Tau22 transgenic model of Alzheimer's disease-like Tau pathology, characterized by the progressive development of Tau pathology, cholinergic alterations and subsequent memory impairments. Three-month-old THY-Tau22 mice and wild-type littermates were assigned to standard housing or housing supplemented with a running wheel. After 9 months of exercise, mice were evaluated for memory performance and examined for hippocampal Tau pathology, cholinergic defects, inflammation and genes related to cholesterol metabolism. Exercise prevented memory alterations in THY-Tau22 mice. This was accompanied by a decrease in hippocampal Tau pathology and a prevention of the loss of expression of choline acetyltransferase within the medial septum. Whereas the expression of most cholesterol-related genes remained unchanged in the hippocampus of running THY-Tau22 mice, we observed a significant upregulation in mRNA levels of NPC1 and NPC2, genes involved in cholesterol trafficking from the lysosomes. Our data support the view that long-term voluntary physical exercise is an effective strategy capable of mitigating Tau pathology and its pathophysiological consequences. Copyright © 2011 Elsevier Inc. All rights reserved.
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Elaboration of antibiofilm surfaces functionalized with antifungal-cyclodextrin inclusion complexes.
Gharbi, Aïcha; Humblot, Vincent; Turpin, Frédéric; Pradier, Claire-Marie; Imbert, Christine; Berjeaud, Jean-Marc
2012-07-01
To tackle the loss of activity of surfaces functionalized by coating and covalently bound molecules to materials, an intermediate system implying the noncovalent immobilization of active molecules in the inner cavity of grafted cyclodextrins (CDs) was investigated. The antifungal and antibiofilm activities of the most stable complexes of Anidulafungin (ANF; echinocandin) and thymol (THY; terpen) in various CDs were demonstrated to be almost the same as the free molecules. The selected CD was covalently bond to self-assembled monolayers on gold surfaces. The immobilized antifungal agents reduced the number of culturable Candida albicans ATCC 3153 attached to the surface by 64 ± 8% for ANF and 75 ± 15% for THY. The inhibitory activity was persistent for THY-loaded samples, whereas it was completely lost for ANF-loaded surfaces after one use. However, reloading of the echinocandin restored the activity. Using fluorescent dying and confocal microscopy, it was proposed that the ANF-loaded surfaces inhibited the adherence of the yeasts, whereas the activity of immobilized THY was found fungicidal. This kind of tailored approach for functionalizing surfaces that could allow a progressive release of ANF or THY gave promising results but still needs to be improved to display a full activity. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.
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Neuroprotective strategies against calpain-mediated neurodegeneration
Yildiz-Unal, Aysegul; Korulu, Sirin; Karabay, Arzu
2015-01-01
Calpains are calcium-dependent proteolytic enzymes that have deleterious effects on neurons upon their pathological over-activation. According to the results of numerous studies to date, there is no doubt that abnormal calpain activation triggers activation and progression of apoptotic processes in neurodegeneration, leading to neuronal death. Thus, it is very crucial to unravel all the aspects of calpain-mediated neurodegeneration in order to protect neurons through eliminating or at least minimizing its lethal effects. Protecting neurons against calpain-activated apoptosis basically requires developing effective, reliable, and most importantly, therapeutically applicable approaches to succeed. From this aspect, the most significant studies focusing on preventing calpain-mediated neurodegeneration include blocking the N-methyl-d-aspartate (NMDA)-type glutamate receptor activities, which are closely related to calpain activation; directly inhibiting calpain itself via intrinsic or synthetic calpain inhibitors, or inhibiting its downstream processes; and utilizing the neuroprotectant steroid hormone estrogen and its receptors. In this review, the most remarkable neuroprotective strategies for calpain-mediated neurodegeneration are categorized and summarized with respect to their advantages and disadvantages over one another, in terms of their efficiency and applicability as a therapeutic regimen in the treatment of neurodegenerative diseases. PMID:25709452
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Know thy sound: perceiving self and others in musical contexts.
Sevdalis, Vassilis; Keller, Peter E
2014-10-01
This review article provides a summary of the findings from empirical studies that investigated recognition of an action's agent by using music and/or other auditory information. Embodied cognition accounts ground higher cognitive functions in lower level sensorimotor functioning. Action simulation, the recruitment of an observer's motor system and its neural substrates when observing actions, has been proposed to be particularly potent for actions that are self-produced. This review examines evidence for such claims from the music domain. It covers studies in which trained or untrained individuals generated and/or perceived (musical) sounds, and were subsequently asked to identify who was the author of the sounds (e.g., the self or another individual) in immediate (online) or delayed (offline) research designs. The review is structured according to the complexity of auditory-motor information available and includes sections on: 1) simple auditory information (e.g., clapping, piano, drum sounds), 2) complex instrumental sound sequences (e.g., piano/organ performances), and 3) musical information embedded within audiovisual performance contexts, when action sequences are both viewed as movements and/or listened to in synchrony with sounds (e.g., conductors' gestures, dance). This work has proven to be informative in unraveling the links between perceptual-motor processes, supporting embodied accounts of human cognition that address action observation. The reported findings are examined in relation to cues that contribute to agency judgments, and their implications for research concerning action understanding and applied musical practice. Copyright © 2014 Elsevier B.V. All rights reserved.
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Enzyme and membrane markers in leukaemia: recent developments.
Hoffbrand, A V; Janossy, G
1981-01-01
Terminal deoxynucleotidyl transferase (TdT) assay has proved a valuable test for distinguishing lymphoblastic from myeloblastic leukaemias, particularly in adults whose blast cells are often negative for the c-ALL antigen. The immunofluorescence assay, particularly when used in combination with antisera to surface membrane antigens, has proved a sensitive technique for detecting small numbers of lymphoblasts in extramedullary sites, for example, testis or cerebrospinal fluid, or of residual Thy-ALL blasts in the marrow, which might otherwise be difficult to recognise. Differences in concentration of several enzymes concerned in purine metabolism have been detected between the blast cells in the various acute leukaemias. Adenosine deaminase (ADA) concentrations tend to be higher in Thy-ALL than in other forms of leukaemia, but the wide overlap reduces the diagnostic value of this assay. Thy-ALL blasts, however, appear to be selectively and exquisitely susceptible to inhibition of ADA by the drug deoxycoformycin, which has now been used sucessfully in a number of other wise resistant patients with Thy-ALL to obtain a complete remission. The recently introduced technique for the production of monoclonal antibodies has substantially widened the reagents available for analysing the membrane characteristics of bone marrow stem cells and of cell lineages derived from them. These have revealed previously unsuspected heterogeneity among different cases of acute lymphoblastic leukaemia, for example, among Thy-ALL blasts from different patients, and they have also delineated minor populations of immature thymocytes from which these leukaemic cells are derived. The potential use of these antibodies to prevent graft-versus-host disease by selective removal of T-lymphocytes from donor bone marrow before allogeneic bone marrow transplantation, or to prevent recurrence of Thy-ALL and other lymphoblastic leukaemias or lymphomas by selective removal of leukaemic or lymphoma malignant cells before autologous transplantation, is reviewed. Images Fig. 2 PMID:6785321
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Glasgow Coma Scale and Outcomes after Structural Traumatic Head Injury in Early Childhood
Heather, Natasha L.; Derraik, José G. B.; Beca, John; Hofman, Paul L.; Dansey, Rangi; Hamill, James; Cutfield, Wayne S.
2013-01-01
Objective To assess the association of the Glasgow Coma Scale (GCS) with radiological evidence of head injury (the Abbreviated Injury Scale for the head region, AIS-HR) in young children hospitalized with traumatic head injury (THI), and the predictive value of GCS and AIS-HR scores for long-term impairment. Methods Our study involved a 10-year retrospective review of a database encompassing all patients admitted to Starship Children’s Hospital (Auckland, New Zealand, 2000–2010) with THI. Results We studied 619 children aged <5 years at the time of THI, with long-term outcome data available for 161 subjects. Both GCS and AIS-HR scores were predictive of length of intensive care unit and hospital stay (all p<0.001). GCS was correlated with AIS-HR (ρ=-0.46; p<0.001), although mild GCS scores (13–15) commonly under-estimated the severity of radiological injury: 42% of children with mild GCS scores had serious–critical THI (AIS-HR 3–5). Increasingly severe GCS or AIS-HR scores were both associated with a greater likelihood of long-term impairment (neurological disability, residual problems, and educational support). However, long-term impairment was also relatively common in children with mild GCS scores paired with structural THI more severe than a simple linear skull fracture. Conclusion Severe GCS scores will identify most cases of severe radiological injury in early childhood, and are good predictors of poor long-term outcome. However, young children admitted to hospital with structural THI and mild GCS scores have an appreciable risk of long-term disability, and also warrant long-term follow-up. PMID:24312648
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Job, Agnès; Cardinal, Fabien; Michel, Hugues; Klein, Céline; Ressiot, Elodie; Gauthier, Jérome
2018-03-26
Tinnitus and associated handicap related to acoustic trauma sequelae have never been assessed in the French artillery. Although impulsive noise exposure to firearms and canons are thought to increase prevalence of tinnitus among soldiers, recent studies demonstrating this fact are missing. Here, a representative sample of 389 soldiers from an operational mountain artillery regiment was surveyed. Soldiers personally concerned by tinnitus were invited to fill in a questionnaire. We assessed tinnitus and the associated handicap using a French translation of the Tinnitus Handicap Inventory (THI). Questions about attention/concentration problems, impaired speech hearing and understanding, sleep disorders, social and familial tension, irritability, depression, and tiredness as linked to tinnitus were the core of the questionnaire. Soldiers that completed the THI (n = 73, 19%) had a mean THI score of 18 ± 17, this mean score corresponded to a mild handicap. At this grade, tinnitus should be easily masked and should not interfere with daily activities. The percentage of soldiers concerned by tinnitus was slightly higher in the older age class, but there was no significant difference of THI scores between the different age classes. The most reported handicaps were attention/concentration problems, impaired speech hearing, and understanding. Among the THI fillers, eight soldiers (11%) had THI scores >36, indicating a moderate to severe handicap. Despite a mild tinnitus handicap, the percentage of people concerned by tinnitus in this regiment is higher (19%) than that in the estimated percentage of general population of European countries (about 10%). It should be of interest to replicate this type of study from other regiments and from other countries. Education and good fitting of hearing protection for prevention of acoustic trauma sequelae should still be encouraged.
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Zheng, Huabao; Wang, Xuan; Yomano, Lorraine P.; Shanmugam, Keelnatham T.
2012-01-01
Furfural is an inhibitory side product formed during the depolymerization of hemicellulose by mineral acids. Genomic libraries from three different bacteria (Bacillus subtilis YB886, Escherichia coli NC3, and Zymomonas mobilis CP4) were screened for genes that conferred furfural resistance on plates. Beneficial plasmids containing the thyA gene (coding for thymidylate synthase) were recovered from all three organisms. Expression of this key gene in the de novo pathway for dTMP biosynthesis improved furfural resistance on plates and during fermentation. A similar benefit was observed by supplementation with thymine, thymidine, or the combination of tetrahydrofolate and serine (precursors for 5,10-methylenetetrahydrofolate, the methyl donor for ThyA). Supplementation with deoxyuridine provided a small benefit, and deoxyribose was of no benefit for furfural tolerance. A combination of thymidine and plasmid expression of thyA was no more effective than either alone. Together, these results demonstrate that furfural tolerance is increased by approaches that increase the supply of pyrimidine deoxyribonucleotides. However, ThyA activity was not directly affected by the addition of furfural. Furfural has been previously shown to damage DNA in E. coli and to activate a cellular response to oxidative damage in yeast. The added burden of repairing furfural-damaged DNA in E. coli would be expected to increase the cellular requirement for dTMP. Increased expression of thyA (E. coli, B. subtilis, or Z. mobilis), supplementation of cultures with thymidine, and supplementation with precursors for 5,10-methylenetetrahydrofolate (methyl donor) are each proposed to increase furfural tolerance by increasing the availability of dTMP for DNA repair. PMID:22504824
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Zheng, Huabao; Wang, Xuan; Yomano, Lorraine P; Shanmugam, Keelnatham T; Ingram, Lonnie O
2012-06-01
Furfural is an inhibitory side product formed during the depolymerization of hemicellulose by mineral acids. Genomic libraries from three different bacteria (Bacillus subtilis YB886, Escherichia coli NC3, and Zymomonas mobilis CP4) were screened for genes that conferred furfural resistance on plates. Beneficial plasmids containing the thyA gene (coding for thymidylate synthase) were recovered from all three organisms. Expression of this key gene in the de novo pathway for dTMP biosynthesis improved furfural resistance on plates and during fermentation. A similar benefit was observed by supplementation with thymine, thymidine, or the combination of tetrahydrofolate and serine (precursors for 5,10-methylenetetrahydrofolate, the methyl donor for ThyA). Supplementation with deoxyuridine provided a small benefit, and deoxyribose was of no benefit for furfural tolerance. A combination of thymidine and plasmid expression of thyA was no more effective than either alone. Together, these results demonstrate that furfural tolerance is increased by approaches that increase the supply of pyrimidine deoxyribonucleotides. However, ThyA activity was not directly affected by the addition of furfural. Furfural has been previously shown to damage DNA in E. coli and to activate a cellular response to oxidative damage in yeast. The added burden of repairing furfural-damaged DNA in E. coli would be expected to increase the cellular requirement for dTMP. Increased expression of thyA (E. coli, B. subtilis, or Z. mobilis), supplementation of cultures with thymidine, and supplementation with precursors for 5,10-methylenetetrahydrofolate (methyl donor) are each proposed to increase furfural tolerance by increasing the availability of dTMP for DNA repair.
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Cirino, Isis Caroline S; Menezes-Silva, Suellen Maria P; Silva, Helena Tainá D; de Souza, Evandro Leite; Siqueira-Júnior, José P
2014-01-01
In an ongoing project to evaluate essential oils as modulators of antibiotic resistance, the essential oil from Origanum vulgare L. (OVEO), as well as its individual constituents carvacrol (CAR) and thymol (THY), were investigated using Staphylococcus aureus strains possessing efflux mechanisms of resistance to norfloxacin, erythromycin and tetracycline. The minimum inhibitory concentration (MIC) values of the antibiotics were determined by agar dilution method, in the absence and in the presence of subinhibitory concentrations of OVEO, CAR or THY. Along with relevant antistaphylococcal activity, OVEO, CAR and THY modulated the activity of tetracycline, i.e. in combination with antibiotics a reduction in the MIC was observed (up to fourfold). The results presented here represent, as far as we know, the first report of OVEO, CAR and THY as putative efflux pump inhibitors. Broadly, these findings indicate that essential oils could serve as potential sources of compounds capable of modulating drug resistance. © 2015 S. Karger AG, Basel.
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Influences of climatic parameters on piglet preweaning mortality in Thailand.
Nuntapaitoon, Morakot; Tummaruk, Padet
2018-04-01
The objective of the present study was to determine the influences of temperature, humidity, and temperature-humidity index (THI) on piglet preweaning mortality in a conventional open-housing system commercial swine herd in Thailand. The analyzed data included 11,157 litters from 3574 Landrace × Yorkshire crossbred sows. The daily temperature, humidity, and THI data were collected from a meteorological station near the herd. The associations between temperature, humidity, and THI for periods before and after farrowings and piglet preweaning mortality were analyzed. Piglet preweaning mortality (log transformation) and the proportion of litters with piglet preweaning mortality greater than 20% were analyzed by using general linear mixed models and generalized linear mixed models (GLIMMIX), respectively. On average, the piglet preweaning mortality and the proportion of litters with piglet preweaning mortality greater than 20% were 14.5% (14.2 to 14.8% CI) and 26.4% (25.5 to 27.2% CI), respectively. Piglet preweaning mortality was positively correlated with the mean temperature (r = 0.028, P = 0.003), humidity (r = 0.038, P < 0.001), and THI (r = 0.036, P < 0.001) during 0-7 days postpartum. In primiparous sows, piglet preweaning mortality increased from 12.1 to 18.5% (+ 6.4%, P < 0.001) when the mean temperature during 0-7 days postpartum increased from < 25.0 to ≥ 29 °C. However, the influence of the temperature during 0-7 days postpartum was insignificant in multiparous sows (P = 0.569, P = 0.593, and P = 0.539 in sows parity numbers 2, 3-5, and 6-9, respectively). Likewise, piglet preweaning mortality increased from 10.7 to 16.7% (+ 6.0%, P = 0.012) when humidity during 0-7 days postpartum increased from < 60 to ≥ 80% in primiparous sows but it was insignificant in sows parity numbers 3-5 (P = 0.095) and 6-9 (P = 0.219). Moreover, the proportion of the litters with piglet preweaning mortality greater than 20% in primiparous sows increased from 18.3 to 32.4% (+ 14.1%, P = 0.017) when the THI during 0-7 days postpartum increased from < 73 to ≥ 81. In conclusion, the negative influences of temperature, humidity, and THI on piglet preweaning mortality were more evident in primiparous than multiparous sows. These findings implied that strategies to reduce temperature for postpartum sows in the open-housing system in Thailand are inadequate, and the proper management of postpartum primiparous sows should be emphasized.
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Min, Li; Cheng, Jian-bo; Shi, Bao-lu; Yang, Hong-jian; Zheng, Nan; Wang, Jia-qi
2015-06-01
Heat stress affects feed intake, milk production, and endocrine status in dairy cows. The temperature-humidity index (THI) is employed as an index to evaluate the degree of heat stress in dairy cows. However, it is difficult to ascertain whether THI is the most appropriate measurement of heat stress in dairy cows. This experiment was conducted to investigate the effects of heat stress on serum insulin, adipokines (leptin and adiponectin), AMP-activated protein kinase (AMPK), and heat shock signal molecules (heat shock transcription factor (HSF) and heat shock proteins (HSP)) in dairy cows and to research biomarkers to be used for better understanding the meaning of THI as a bioclimatic index. To achieve these objectives, two experiments were performed. The first experiment: eighteen lactating Holstein dairy cows were used. The treatments were: heat stress (HS, THI average=81.7, n=9) and cooling (CL, THI average=53.4, n=9). Samples of HS were obtained on August 16, 2013, and samples of CL were collected on April 7, 2014 in natural conditions. The second experiment: HS treatment cows (n=9) from the first experiment were fed for 8 weeks from August 16, 2013 to October 12, 2013. Samples for moderate heat stress, mild heat stress, and no heat stress were obtained, respectively, according to the physical alterations of the THI. Results showed that heat stress significantly increased the serum adiponectin, AMPK, HSF, HSP27, HSP70, and HSP90 (P<0.05). Adiponectin is strongly associated with AMPK. The increases of adiponectin and AMPK may be one of the mechanisms to maintain homeostasis in heat-stressed dairy cows. When heat stress treatment lasted 8 weeks, a higher expression of HSF and HSP70 was observed under moderate heat stress. Serum HSF and HSP70 are sensitive and accurate in heat stress and they could be potential indicators of animal response to heat stress. We recommend serum HSF and HSP70 as meaningful biomarkers to supplement the THI and evaluate moderate heat stress in dairy cows in the future.
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Min, Li; Cheng, Jian-bo; Shi, Bao-lu; Yang, Hong-jian; Zheng, Nan; Wang, Jia-qi
2015-01-01
Heat stress affects feed intake, milk production, and endocrine status in dairy cows. The temperature-humidity index (THI) is employed as an index to evaluate the degree of heat stress in dairy cows. However, it is difficult to ascertain whether THI is the most appropriate measurement of heat stress in dairy cows. This experiment was conducted to investigate the effects of heat stress on serum insulin, adipokines (leptin and adiponectin), AMP-activated protein kinase (AMPK), and heat shock signal molecules (heat shock transcription factor (HSF) and heat shock proteins (HSP)) in dairy cows and to research biomarkers to be used for better understanding the meaning of THI as a bioclimatic index. To achieve these objectives, two experiments were performed. The first experiment: eighteen lactating Holstein dairy cows were used. The treatments were: heat stress (HS, THI average=81.7, n=9) and cooling (CL, THI average=53.4, n=9). Samples of HS were obtained on August 16, 2013, and samples of CL were collected on April 7, 2014 in natural conditions. The second experiment: HS treatment cows (n=9) from the first experiment were fed for 8 weeks from August 16, 2013 to October 12, 2013. Samples for moderate heat stress, mild heat stress, and no heat stress were obtained, respectively, according to the physical alterations of the THI. Results showed that heat stress significantly increased the serum adiponectin, AMPK, HSF, HSP27, HSP70, and HSP90 (P<0.05). Adiponectin is strongly associated with AMPK. The increases of adiponectin and AMPK may be one of the mechanisms to maintain homeostasis in heat-stressed dairy cows. When heat stress treatment lasted 8 weeks, a higher expression of HSF and HSP70 was observed under moderate heat stress. Serum HSF and HSP70 are sensitive and accurate in heat stress and they could be potential indicators of animal response to heat stress. We recommend serum HSF and HSP70 as meaningful biomarkers to supplement the THI and evaluate moderate heat stress in dairy cows in the future. PMID:26055916
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Brovarets', Ol'ha O; Zhurakivsky, Roman O; Hovorun, Dmytro M
2013-10-01
The biologically important tautomerization of the Hyp·Cyt, Hyp·Thy and Hyp·Hyp base pairs to the Hyp·Cyt, Hyp·Thy and Hyp·Hyp base pairs, respectively, by the double proton transfer (DPT) was comprehensively studied in vacuo and in the continuum with a low dielectric constant (ε = 4) corresponding to hydrophobic interfaces of protein-nucleic acid interactions by combining theoretical investigations at the B3LYP/6-311++G(d,p) level of QM theory with QTAIM topological analysis. Based on the sweeps of the energetic, electron-topological, geometric and polar parameters, which describe the course of the tautomerization along the intrinsic reaction coordinate (IRC), it was proved that the tautomerization through the DPT is concerted and asynchronous process for the Hyp·Cyt and Hyp·Thy base pairs, while concerted and synchronous for the Hyp·Hyp homodimer. The continuum with ε = 4 does not affect qualitatively the course of the tautomerization reaction for all studied complexes. The nine key points along the IRC of the Hyp·Cyt↔Hyp·Cyt and Hyp·Thy↔Hyp·Thy tautomerizations and the six key points of the Hyp·Hyp↔Hyp·Hyp tautomerization have been identified and fully characterized. These key points could be considered as electron-topological "fingerprints" of concerted asynchronous (for Hyp·Cyt and Hyp·Thy) or synchronous (for Hyp·Hyp) tautomerization process via the DPT. It was found, that in the Hyp·Cyt, Hyp·Thy, Hyp·Hyp and Hyp·Hyp base pairs all H-bonds are significantly cooperative and mutually reinforce each other, while the C2H…O2 H-bond in the Hyp·Cyt base pair and the O6H…O4 H-bond in the Hyp·Thy base pair behave anti-cooperatively, i.e., they become weakened, while two others become strengthened.
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U.S. EPA, Pesticide Product Label, MALEIC HYDRAZIDE 2P, 11/21/1972
2011-04-14
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Effect of heat stress on age at first calving of Japanese Black cows in Okinawa.
Oikawa, Takuro
2017-03-01
Calving records from birth certificates of cows were analyzed to investigate the effect of heat stress on age at first calving (AFC) of Japanese Black cows. The data set covered 20 years (1990-2009) of calving records. Total number of records was 9279. Daily weather information from weather stations in the vicinity of the farms was used. Temperature-humidity index (THI) fitted to a linear model covered 30 days pre-insemination to 61 days post-insemination. Statistical analysis was conducted with procedures of SAS/STAT. Preliminary analysis showed that THI of the lowest temperature and humidity was most conducive to AFC. Covariance analysis, including main effect of sire, farm and year of insemination and covariates of THI on days showed that regression coefficients of THI on day -7, day -2 and day +31 were statistically significant. The estimated piecewise regression line showed different responses of AFC to THI on days: roof-shasped downward trend on day -7, hockey-stick shaped upward trend on day -2 and day +31. The difference among the estimated regression lines may be caused by direct and indirect factors on reproduction: indirect effect of reduced feed intake, failure of conception at previous insemination, direct effect of heat stress on oocyte and embryo development. © 2016 Japanese Society of Animal Science.
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Increased Atherosclerosis Correlates with Subjective Tinnitus Severity.
Yüksel, Fatih; Karataş, Duran; Türkdoğan, Figen Tunalı; Yüksel, Özlem
2018-03-01
The aim of the present study was to investigate whether increased intima media thickness was associated with the severity of subjective non-pulsatile tinnitus and hearing loss. Data of the patients who came to Otorhinolaryngology Department of Isparta Government Hospital with subjective non-pulsatile tinnitus complaint, between January 2012 and June 2013, were evaluated retrospectively. A total of 215 patients were included in the present study. Hearing tests, biochemical analysis, tinnitus handicap inventory (THI), visual analogue scale (VAS) and doppler ultrasonography results of the patients were reviewed and recorded. The patients were classified into two groups as those having an increased intima media thickness and those having a normal intima media thickness. The said groups were compared with respect to age, gender, THI, VAS, hearing test findings and lipid values. Moreover, THI and VAS groups were compared with respect to intima-media thickness. In the group having increased intima-media thickness, THI and VAS average, frequency of hypertension, total cholesterol, low density lipoprotein and triglyceride averages and mean frequencies obtained by hearing test were significantly higher. Comparison of THI and VAS groups showed that intima-media thickness was significantly different between those having a mild tinnitus and those having a severe tinnitus. Increased intima-media thickness was associated with the severity of subjective non-pulsatile tinnitus and hearing loss. For this reason, the carotid system should be examined in subjective non-pulsatile tinnitus patients.
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Derix, Johanna; Iljina, Olga; Weiske, Johanna; Schulze-Bonhage, Andreas; Aertsen, Ad; Ball, Tonio
2014-01-01
Exchange of thoughts by means of expressive speech is fundamental to human communication. However, the neuronal basis of real-life communication in general, and of verbal exchange of ideas in particular, has rarely been studied until now. Here, our aim was to establish an approach for exploring the neuronal processes related to cognitive “idea” units (IUs) in conditions of non-experimental speech production. We investigated whether such units corresponding to single, coherent chunks of speech with syntactically-defined borders, are useful to unravel the neuronal mechanisms underlying real-world human cognition. To this aim, we employed simultaneous electrocorticography (ECoG) and video recordings obtained in pre-neurosurgical diagnostics of epilepsy patients. We transcribed non-experimental, daily hospital conversations, identified IUs in transcriptions of the patients' speech, classified the obtained IUs according to a previously-proposed taxonomy focusing on memory content, and investigated the underlying neuronal activity. In each of our three subjects, we were able to collect a large number of IUs which could be assigned to different functional IU subclasses with a high inter-rater agreement. Robust IU-onset-related changes in spectral magnitude could be observed in high gamma frequencies (70–150 Hz) on the inferior lateral convexity and in the superior temporal cortex regardless of the IU content. A comparison of the topography of these responses with mouth motor and speech areas identified by electrocortical stimulation showed that IUs might be of use for extraoperative mapping of eloquent cortex (average sensitivity: 44.4%, average specificity: 91.1%). High gamma responses specific to memory-related IU subclasses were observed in the inferior parietal and prefrontal regions. IU-based analysis of ECoG recordings during non-experimental communication thus elicits topographically- and functionally-specific effects. We conclude that segmentation of spontaneous real-world speech in linguistically-motivated units is a promising strategy for elucidating the neuronal basis of mental processing during non-experimental communication. PMID:24982625
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Constraining the Teratogenicity of Pesticide Pollution by a Synthetic Nanoreceptor.
Yang, Xue; Li, Shengke; Wang, Ziyi; Lee, Simon M Y; Wang, Lian-Hui; Wang, Ruibing
2018-01-04
The teratogenicity of the pesticide nereistoxin (NTX) and its derivative thiocyclam (THI) towards aquatic life was dramatically constrained by a synthetic nanoreceptor, cucurbit[7]uril, through selective encapsulation of the pesticides (K CB[7]-NTX of 3.24(±0.31)×10 6 m -1 and K CB[7]-THI of 7.46(±0.10)×10 5 m -1 ), as evidenced by the rate of hatchability, morphology development, and tyrosinase activity of zebrafish larvae incubated with the pesticides (3-300 μm) in the absence and in the presence of 300 μm cucurbit[7]uril, demonstrating the significant potential of the nanoreceptor in managing ecological pollution of these pesticides. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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van den Berge, Minke J C; van Dijk, J Marc C; Free, Rolien H; Stienstra, Janke; van Dijk, Pim; van der Laan, Bernard F A M
2017-02-01
Tinnitus is a common entity that may lead to severe impairment in quality of life. An adequate treatment modality for severe tinnitus is currently lacking. Neurostimulation of the auditory tract may serve as a promising adjunct in tinnitus treatment. The aim is to investigate the effect of direct stimulation on the cochleovestibular nerve for intractable tinnitus. This study was conducted at the University Medical Center Groningen, The Netherlands. We studied 10 patients with severe, unilateral, intractable tinnitus, who were implanted with a cuff electrode around the cochleovestibular nerve between 2001 and 2013. All patients had preoperative ipsilateral hearing loss. Tinnitus Handicap Inventory (THI) scores and audiometric values were collected. Treatment success was determined based on the self-assessment of satisfactory usage by each patient. The mean preoperative tinnitus duration was 8.0 ± 5.9 years. The preoperative THI score was 71 ± 18 points. During mean follow-up of 49 months, the mean THI reduction was 24 ± 26 points (P = 0.02). Treatment was regarded successful in 6 patients (60%). In these patients, tinnitus did not disappear, but transformed into a more bearable sound. In 4 patients, transient complications occurred, and 1 patient experienced permanent vertigo postoperatively. Furthermore, hearing deterioration was a result of implantation in 86% of the patients. Direct neurostimulation resulted in treatment success in a small majority of the patients, with a significant decrease in THI score. However, because of a high risk of additional hearing damage, this technique seems not viable for patients with moderate hearing loss. Copyright © 2016 Elsevier Inc. All rights reserved.
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Tinnitus and Its Effect on the Quality of Life of Sufferers: A Nigerian Cohort Study.
Ukaegbe, Onyinyechi C; Orji, Foster T; Ezeanolue, Basil C; Akpeh, James O; Okorafor, Ijeoma A
2017-10-01
Objectives To evaluate the quality of life of patients with ongoing tinnitus. Study Design This was a cross-sectional study of patients with ongoing tinnitus. Setting The study was carried out in a tertiary hospital in southeastern Nigeria. Subjects and Methods Subjects are adults who presented to the otorhinolaryngology clinic with tinnitus as their primary complaint. Pure-tone audiometry, tinnitus pitch, and loudness matching were done. The Tinnitus Handicap Inventory (THI) questionnaire was used in assessing their quality of life. Results There were 63 participants within the age range of 16 to 74 years; 20 (31.7%) were male and 43 (68.3%) were female. The mean duration of tinnitus was 26.7 ± 38.1 months. Nineteen (30.2%) participants had bilateral tinnitus while 44 (69.8%) had unilateral tinnitus. The mean THI score was 36.6 ± 19.7. The most reported handicap was anxiety and difficulty with concentration followed by depression and irritability. There was no correlation between the disability shown by the THI score and the age, sex, duration of the tinnitus, the tinnitus pitch, tinnitus loudness, or the laterality of the tinnitus. There was a significant positive correlation between the grade of hearing loss and the level of disability reported in the THI ( P = .01). Conclusion Tinnitus sufferers appear to have poorer quality of life compared with nonsufferers. This quality-of-life affectation is likely to be worse in those with disabling hearing loss but does not appear to be related to their age, sex, symptom duration, or the loudness and pitch of their tinnitus.
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Serotonin-containing neurons in basal insects: In search of ground patterns among tetraconata.
Stemme, Torben; Stern, Michael; Bicker, Gerd
2017-01-01
The ventral nerve cord of Tetraconata contains a comparably low number of serotonin-immunoreactive neurons, facilitating individual identification of cells and their characteristic neurite morphology. This offers the rather unique possibility of establishing homologies at the single cell level. Because phylogenetic relationships within Tetraconata are still discussed controversially, comparisons of individually identifiable neurons can help to unravel these issues. Serotonin immunoreactivity has been investigated in numerous tetraconate taxa, leading to reconstructions of hypothetical ground patterns for major lineages. However, detailed descriptions of basal insects are still missing, but are crucial for meaningful evolutionary considerations. We investigated the morphology of individually identifiable serotonin-immunoreactive neurons in the ventral nerve cord of Zygentoma (Thermobia domestica, Lepisma saccharina, Atelura formicaria) and Archaeognatha (Machilis germanica, Dilta hibernica). To improve immunocytochemical resolution, we also performed preincubation experiments with 5-hydroxy-L-tryptophan and serotonin. Additionally, we checked for immunolabeling of tryptophan hydroxylase, an enzyme associated with the synthesis of serotonin. Besides the generally identified groups of anterolateral, medial, and posterolateral neurons within each ganglion of the ventral nerve cord, we identified several other immunoreactive cells, which seem to have no correspondence in other tetraconates. Furthermore, we show that not all immunoreactive neurons produce serotonin, but have the capability for serotonin uptake. Comparisons with the patterns of serotonin-containing neurons in major tetraconate taxa suggest a close phylogenetic relationship of Remipedia, Cephalocarida, and Hexapoda, supporting the Miracrustacea hypothesis. J. Comp. Neurol., 2016. © 2016 Wiley Periodicals, Inc. J. Comp. Neurol. 525:79-115, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.
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Hua, XinZhong; Hua, LinQiang; Liu, XiaoJun
2016-05-18
The nonradiative decay dynamics of photo-excited uracil (Ura) and its derivatives, i.e., thymine (5-methyluracil, Thy), 6-methyluracil (6-MU) and 6-azauracil (6-AU) in water, has been studied using a femtosecond transient absorption method. The molecules are populated in the lowest (1)ππ* state by a pump pulse at 266 nm, and a broadband continuum in the deep UV region is then employed as the probe. The extension of the continuous UV probe down to 250 nm enables us to investigate comprehensively the population dynamics of the ground states for those molecules and to uncover the substituent effects on nonradiative decay dynamics of uracil. Vibrational cooling in the ground states of Ura, Thy and 6-MU has been directly observed for the first time, providing solid evidence of the ultrafast (1)ππ* → S0 decay. In combination with the ground state bleaching signals, it is consolidated that their lowest (1)ππ* state decays via two parallel pathways, i.e., (1)ππ* → S0 and (1)ππ* → (1)nπ*. Moreover, the contribution of the (1)ππ* → (1)nπ* channel is found to be much smaller for Thy or 6-MU than for Ura. Different from methyl-substitution, the initial (1)ππ* state of the aza-substituent 6-AU decays primarily to the (1)nπ* state, while the (1)ππ* → S0 channel can be negligible. Our study provides a comprehensive understanding of the substituent effects on the excited-state dynamics of uracil in water.
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Endomicroscopy and electromyography of neuromuscular junctions in situ
Brown, Rosalind; Dissanayake, Kosala N; Skehel, Paul A; Ribchester, Richard R
2014-01-01
Objective Electromyography (EMG) is used routinely to diagnose neuromuscular dysfunction in a wide range of peripheral neuropathies, myopathies, and neuromuscular degenerative diseases including motor neuron diseases such as amyotrophic lateral sclerosis (ALS). Definitive neurological diagnosis may also be indicated by the analysis of pathological neuromuscular innervation in motor-point biopsies. Our objective in this study was to preempt motor-point biopsy by combining live imaging with electrophysiological analysis of slow degeneration of neuromuscular junctions (NMJs) in vivo. Methods We combined conventional needle electromyography with fiber-optic confocal endomicroscopy (CEM), using an integrated hand-held, 1.5-mm-diameter probe. We utilized as a test bed, various axotomized muscles in the hind limbs of anaesthetized, double-homozygous thy1.2YFP16: WldS mice, which coexpress the Wallerian-degeneration Slow (WldS) protein and yellow fluorescent protein (YFP) in motor neurons. We also tested exogenous vital stains, including Alexa488-α-bungarotoxin; the styryl pyridinium dye 4-Di-2-Asp; and a GFP conjugate of botulinum toxin Type A heavy chain (GFP-HcBoNT/A). Results We show that an integrated EMG/CEM probe is effective in longitudinal evaluation of functional and morphological changes that take place over a 7-day period during axotomy-induced, slow neuromuscular synaptic degeneration. EMG amplitude declined in parallel with overt degeneration of motor nerve terminals. EMG/CEM was safe and effective when nerve terminals and motor endplates were selectively stained with vital dyes. Interpretation Our findings constitute proof-of-concept, based on live imaging in an animal model, that combining EMG/CEM may be useful as a minimally invasive precursor or alternative to motor-point biopsy in neurological diagnosis and for monitoring local administration of potential therapeutics. PMID:25540801
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Khalaj, Leila; Nejad, Sara Chavoshi; Mohammadi, Marzieh; Zadeh, Sadaf Sarraf; Pour, Marieh Hossein; Ahmadiani, Abolhassan; Khodagholi, Fariba; Ashabi, Ghorbangol; Alamdary, Shabnam Zeighamy; Samami, Elham
2013-08-21
Stress predisposes the brain to various neuropathological disorders. Fibrates like gemfibrozil, commonly used for hyperlipidemia, have not yet been examined for their protective/deteriorative potential against restraint stress-induced disturbances. Pretreatment of rats with a range of gemfibrozil concentrations showed significant protection against stress consequences at 90 mg/kg of gemfibrozil, as it resulted in the highest level of antioxidant defense system potentiation among other doses. It also reduced plasma corticosterone compared with the stressed animals. Administration of gemfibrozil (90 mg/kg) before stress induction was able to significantly induce the protein levels of some protective factors including hemeoxygenase-1 (HO-1) and NAD(P)H dehydrogenase quinone-1 (NQO-1) in the antioxidant nuclear factor erythroid-derived 2-like 2 (Nrf-2) pathway, as well as mitochondrial pro-survival proteins, including peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and nuclear respiratory factor 1 (NRF-1). In parallel, the level of cleaved caspase-3 and apoptosis-inducing factor (AIF), two proteins involved in apoptotic cell death, and the number of damaged neurons detected in hematoxylin-eosin (H&E) stained hippocampus sections were suppressed in the presence of gemfibrozil. Herein, although gemfibrozil demonstrated protection against the restraint stress, considering its dose and context-dependent effects reported in the previous studies, as well as its common application in clinic, further investigations are essential to unravel its exact beneficial/deleterious effects in various neuronal contexts. Copyright © 2013 Elsevier B.V. All rights reserved.
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Auditory discrimination training for tinnitus treatment: the effect of different paradigms.
Herraiz, Carlos; Diges, I; Cobo, P; Aparicio, J M; Toledano, A
2010-07-01
Acoustic deprivation, i.e. hearing loss, is responsible for a cascade of processes resulting in reorganisation of the cortex. Tinnitus mechanisms are explained by synchronization of the neural spontaneous activity and might be related to cortical re-mapping. Auditory discrimination training (ADT) has demonstrated in both animals and humans to induce tonotopical changes in the auditory pathways through neural plasticity. We hypothesize that ADT could have some effect on tinnitus perception. The objective of this study is to compare the effect on tinnitus following two paradigms of ADT. Only patients from 20 to 60 years of age were recruited. Inclusion criteria were pure tone tinnitus of mild or moderate handicap according to the Tinnitus Handicap Inventory score (<56). ADT patients were randomized in two groups: SAME (ADT in the same frequency of tinnitus pitch, 20 patients) and NONSAME (ADT in the frequency one-octave below tinnitus pitch, 21 patients). Groups of pair of tones (70% standard tones ST, 30% deviant tones ST + 0.1-0.5 kHz) were randomly mixed for 20 min/day during 1 month. Patient had to mark when the two sounds of the pair were similar or different. Control group included 26 patients from the waiting list (WLG). Patients were also divided according to the trained frequency and the deepest hearing-impaired frequency. Outcome parameters were set up according to the answer to the question "is your tinnitus better, same, or worse with the treatment?" (RESP), the tinnitus handicap inventory (THI) and the visual analogue scale from 1 to 10 on tinnitus intensity (VAS). Tinnitus improved in 42.2% of the patients (RESP). VAS and THI scores were reduced but only THI differences were statistically significant (P = 0.003). ADT patients improved significantly compared with WLG in RESP and THI scores (P < 0.01). Training frequencies one-octave below the tinnitus pitch (NONSAME) decreased significantly THI scores compared with patients trained frequencies similar to tinnitus pitch (SAME, P = 0.035). RESP and VAS scores decreased more in NONSAME group though differences were not significant. We did not find any differences when comparing the group training the deepest hearing-impaired frequency and the group who trained other frequencies. Auditory discrimination training significantly improved tinnitus handicap compared to a waiting list group. Those patients who trained frequencies one octave below the tinnitus pitch had better outcome than those who performed the ADT with frequencies similar to the tinnitus pitch (P = 0.035).
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Banerjee, Surajit; Christov, Plamen P.; Kozekova, Albena
trans-4-Hydroxynonenal (HNE) is the major peroxidation product of {omega}-6 polyunsaturated fatty acids in vivo. Michael addition of the N{sub 2}-amino group of dGuo to HNE followed by ring closure of N1 onto the aldehyde results in four diastereomeric 1,N{sub 2}-dGuo (1,N{sub 2}-HNE-dGuo) adducts. The (6S,8R,11S)-HNE-1,N{sub 2}-dGuo adduct was incorporated into the 18-mer templates 5'-d(TCATXGAATCCTTCCCCC)-3' and d(TCACXGAATCCTTCCCCC)-3', where X = (6S,8R,11S)-HNE-1,N{sub 2}-dGuo adduct. These differed in the identity of the template 5'-neighbor base, which was either Thy or Cyt, respectively. Each of these templates was annealed with either a 13-mer primer 5'-d(GGGGGAAGGATTC)-3' or a 14-mer primer 5'-d(GGGGGAAGGATTCC)-3'. The addition of dNTPsmore » to the 13-mer primer allowed analysis of dNTP insertion opposite to the (6S,8R,11S)-HNE-1,N{sub 2}-dGuo adduct, whereas the 14-mer primer allowed analysis of dNTP extension past a primed (6S,8R,11S)-HNE-1,N{sub 2}-dGuo:dCyd pair. The Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4) belongs to the Y-family of error-prone polymerases. Replication bypass studies in vitro reveal that this polymerase inserted dNTPs opposite the (6S,8R,11S)-HNE-1,N{sub 2}-dGuo adduct in a sequence-specific manner. If the template 5'-neighbor base was dCyt, the polymerase inserted primarily dGTP, whereas if the template 5'-neighbor base was dThy, the polymerase inserted primarily dATP. The latter event would predict low levels of Gua {yields} Thy mutations during replication bypass when the template 5'-neighbor base is dThy. When presented with a primed (6S,8R,11S)-HNE-1,N{sub 2}-dGuo:dCyd pair, the polymerase conducted full-length primer extension. Structures for ternary (Dpo4-DNA-dNTP) complexes with all four template-primers were obtained. For the 18-mer:13-mer template-primers in which the polymerase was confronted with the (6S,8R,11S)-HNE-1,N{sub 2}-dGuo adduct, the (6S,8R,11S)-1,N{sub 2}-dGuo lesion remained in the ring-closed conformation at the active site. The incoming dNTP, either dGTP or dATP, was positioned with Watson-Crick pairing opposite the template 5'-neighbor base, dCyt or dThy, respectively. In contrast, for the 18-mer:14-mer template-primers with a primed (6S,8R,11S)-HNE-1,N{sub 2}-dGuo:dCyd pair, ring opening of the adduct to the corresponding N{sub 2}-dGuo aldehyde species occurred. This allowed Watson-Crick base pairing at the (6S,8R,11S)-HNE-1,N{sub 2}-dGuo:dCyd pair.« less
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Pavani, Krishna; Carvalhais, Isabel; Faheem, Marwa; Chaveiro, Antonio; Reis, Francisco Vieira; da Silva, Fernando Moreira
2015-01-01
The present study was designed to evaluate how environmental factors in a dry-summer subtropical climate in Terceira-Azores (situated in the North Atlantic Ocean: 38° 43′ N 27° 12′ W) can affect dairy cow (Holstein) fertility, as well as seasonal influence on in vitro oocytes maturation and embryos development. Impact of heat shock (HS) effects on in vitro oocyte’s maturation and further embryo development after in vitro fertilization (IVF) was also evaluated. For such purpose the result of the first artificial insemination (AI) performed 60 to 90 days after calving of 6,300 cows were recorded for one year. In parallel, climatic data was obtained at different elevation points (n = 5) from 0 to 1,000 m and grazing points from 0 to 500 m, in Terceira island, and the temperature humidity index (THI) was calculated. For in vitro experiments, oocytes (n = 706) were collected weekly during all year, for meiotic maturation and IVF. Further, to evaluate HS effect, 891 oocytes were collected in the cold moths (December, January, February and March) and divided in three groups treated to HS for 24 h during in vitro maturation at: C (Control = 38.5°C), HS1 (39.5°C) and HS2 (40.5°C). Oocytes from each group were used for meiotic assessment and IVF. Cleavage, morula and blastocyst development were evaluated respectively on day 2, 6, and 9 after IVF. A negative correlation between cow’s conception rate (CR) and THI in grazing points (−91.3%; p<0.001) was observed. Mean THI in warmer months (June, July, August and September) was 71.7±0.7 and the CR (40.2±1.5%) while in cold months THI was 62.8±0.2 and CR was 63.8±0.4%. A similar impact was obtained with in vitro results in which nuclear maturation rate (NMR) ranged from 78.4% (±8.0) to 44.3% (±8.1), while embryos development ranged from 53.8% (±5.8) to 36.3% (±3.3) in cold and warmer months respectively. In vitro HS results showed a significant decline (p<0.05) on NMR of oocytes for every 1°C rising temperature (78.4±8.0, 21.7±3.1 and 8.9±2.2, respectively for C, HS1, and HS2). Similar results were observed in cleavage rate and embryo development, showing a clear correlation (96.9 p<0.05) between NMR and embryo development with respect to temperatures. Results clearly demonstrated that, up to a THI of 70.6, a decrease in the CR occurs in first AI after calving; this impairment was confirmed with in vitro results. PMID:25656191
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Pavani, Krishna; Carvalhais, Isabel; Faheem, Marwa; Chaveiro, Antonio; Reis, Francisco Vieira; da Silva, Fernando Moreira
2015-03-01
The present study was designed to evaluate how environmental factors in a dry-summer subtropical climate in Terceira-Azores (situated in the North Atlantic Ocean: 38° 43' N 27° 12' W) can affect dairy cow (Holstein) fertility, as well as seasonal influence on in vitro oocytes maturation and embryos development. Impact of heat shock (HS) effects on in vitro oocyte's maturation and further embryo development after in vitro fertilization (IVF) was also evaluated. For such purpose the result of the first artificial insemination (AI) performed 60 to 90 days after calving of 6,300 cows were recorded for one year. In parallel, climatic data was obtained at different elevation points (n = 5) from 0 to 1,000 m and grazing points from 0 to 500 m, in Terceira island, and the temperature humidity index (THI) was calculated. For in vitro experiments, oocytes (n = 706) were collected weekly during all year, for meiotic maturation and IVF. Further, to evaluate HS effect, 891 oocytes were collected in the cold moths (December, January, February and March) and divided in three groups treated to HS for 24 h during in vitro maturation at: C (Control = 38.5°C), HS1 (39.5°C) and HS2 (40.5°C). Oocytes from each group were used for meiotic assessment and IVF. Cleavage, morula and blastocyst development were evaluated respectively on day 2, 6, and 9 after IVF. A negative correlation between cow's conception rate (CR) and THI in grazing points (-91.3%; p<0.001) was observed. Mean THI in warmer months (June, July, August and September) was 71.7±0.7 and the CR (40.2±1.5%) while in cold months THI was 62.8±0.2 and CR was 63.8±0.4%. A similar impact was obtained with in vitro results in which nuclear maturation rate (NMR) ranged from 78.4% (±8.0) to 44.3% (±8.1), while embryos development ranged from 53.8% (±5.8) to 36.3% (±3.3) in cold and warmer months respectively. In vitro HS results showed a significant decline (p<0.05) on NMR of oocytes for every 1°C rising temperature (78.4±8.0, 21.7±3.1 and 8.9±2.2, respectively for C, HS1, and HS2). Similar results were observed in cleavage rate and embryo development, showing a clear correlation (96.9 p<0.05) between NMR and embryo development with respect to temperatures. Results clearly demonstrated that, up to a THI of 70.6, a decrease in the CR occurs in first AI after calving; this impairment was confirmed with in vitro results.
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Pulsed Beamless High Power Microwave (HPM) Source with Integrated Antenna
2013-06-01
lengt ance matchin ces have bee ntenna. Thi self-containe 150-550 MH power of thi m with a puls MICRO ED ANTE tromagnetic eign Trade ltgilbers U z...nal at ith a igned, ng of ve the 0-550 a. The ed by dance lowed peak ically- tional efined 88. 330
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Tinnitus severity, depression, and the big five personality traits.
Langguth, B; Kleinjung, T; Fischer, B; Hajak, G; Eichhammer, P; Sand, P G
2007-01-01
A growing number of self-report measures for the evaluation of tinnitus severity has become available to research and clinical practice. This has led to an increased awareness of depression and personality as predictors of tinnitus severity in addition to loudness and other psychoacoustic measures. However, the net impact of personality dimensions on tinnitus ratings has not been investigated when the effect of depressed mood is controlled. In the present study, we demonstrate the role of the big five personality traits, 'Neuroticism', 'Extraversion', 'Openness', 'Agreeableness', and 'Conscientiousness', in affecting scores on two standard instruments for grading tinnitus-related complaints, the tinnitus handicap inventory (THI), and the tinnitus questionnaire (TQ). When 72 individuals with chronic tinnitus were examined, 'Agreeableness' negatively correlated with THI scores (p=.003), whereas the anxiety trait 'Neuroticism' correlated both with depressive symptomatology (p<.001) and TQ scores (p=.028), but not with THI ratings (n.s.). In addition to confirming the established roles of trait anxiety and depression, low 'Agreeableness' was thus identified as a novel predictor of tinnitus severity on the THI.
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Conformational changes in the expression domain of the Escherichia coli thiM riboswitch
Rentmeister, Andrea; Mayer, Günter; Kuhn, Nicole; Famulok, Michael
2007-01-01
The thiM riboswitch contains an aptamer domain that adaptively binds the coenzyme thiamine pyrophosphate (TPP). The binding of TPP to the aptamer domain induces structural rearrangements that are relayed to a second domain, the so-called expression domain, thereby interfering with gene expression. The recently solved crystal structures of the aptamer domains of the thiM riboswitches in complex with TPP revealed how TPP stabilizes secondary and tertiary structures in the RNA ligand complex. To understand the global modes of reorganization between the two domains upon metabolite binding the structure of the entire riboswitch in presence and absence of TPP needs to be determined. Here we report the secondary structure of the entire thiM riboswitch from Escherichia coli in its TPP-free form and its transition into the TPP-bound variant, thereby depicting domains of the riboswitch that serve as communication links between the aptamer and the expression domain. Furthermore, structural probing provides an explanation for the lack of genetic control exerted by a riboswitch variant with mutations in the expression domain that still binds TPP. PMID:17517779
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Low heat transfer oxidizer heat exchanger design and analysis
NASA Technical Reports Server (NTRS)
Kanic, P. G.; Kmiec, T. D.; Peckham, R. J.
1987-01-01
The RL10-IIB engine, a derivative of the RLIO, is capable of multi-mode thrust operation. This engine operates at two low thrust levels: tank head idle (THI), which is approximately 1 to 2 percent of full thrust, and pumped idle (PI), which is 10 percent of full thrust. Operation at THI provides vehicle propellant settling thrust and efficient engine thermal conditioning; PI operation provides vehicle tank pre-pressurization and maneuver thrust for log-g deployment. Stable combustion of the RL10-IIB engine at THI and PI thrust levels can be accomplished by providing gaseous oxygen at the propellant injector. Using gaseous hydrogen from the thrust chamber jacket as an energy source, a heat exchanger can be used to vaporize liquid oxygen without creating flow instability. This report summarizes the design and analysis of a United Aircraft Products (UAP) low-rate heat transfer heat exchanger concept for the RL10-IIB rocket engine. The design represents a second iteration of the RL10-IIB heat exchanger investigation program. The design and analysis of the first heat exchanger effort is presented in more detail in NASA CR-174857. Testing of the previous design is detailed in NASA CR-179487.
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Clonal development and organization of the adult Drosophila central brain.
Yu, Hung-Hsiang; Awasaki, Takeshi; Schroeder, Mark David; Long, Fuhui; Yang, Jacob S; He, Yisheng; Ding, Peng; Kao, Jui-Chun; Wu, Gloria Yueh-Yi; Peng, Hanchuan; Myers, Gene; Lee, Tzumin
2013-04-22
The insect brain can be divided into neuropils that are formed by neurites of both local and remote origin. The complexity of the interconnections obscures how these neuropils are established and interconnected through development. The Drosophila central brain develops from a fixed number of neuroblasts (NBs) that deposit neurons in regional clusters. By determining individual NB clones and pursuing their projections into specific neuropils, we unravel the regional development of the brain neural network. Exhaustive clonal analysis revealed 95 stereotyped neuronal lineages with characteristic cell-body locations and neurite trajectories. Most clones show complex projection patterns, but despite the complexity, neighboring clones often coinnervate the same local neuropil or neuropils and further target a restricted set of distant neuropils. These observations argue for regional clonal development of both neuropils and neuropil connectivity throughout the Drosophila central brain. Copyright © 2013 Elsevier Ltd. All rights reserved.
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Danaei, Gholam Hassan; Memar, Bahram; Ataee, Ramin; Karami, Mohammad
2018-04-12
Several studies have shown that oxidative stress and cell damage can occur at very early stages of diazinon (DZN) exposure. The present study was designed to determine the beneficial effect of thymoquinone (Thy), the main component of Nigella sativa (black seed or black cumin), against DZN cardio-toxicity in rats. In the present experimental study, 48 male Wistar rats were randomly divided into six groups: control (corn oil gavages), DZN gavages (20 mg/kg/day), Thy gavages (10 mg/kg/day) and Thy + DVN gavages (2.5, 5 and 10 mg/kg/day). Treatments were continued for 28 days, then the animals were anesthetized by ether and superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), lactate dehydrogenize (LDH) and glutathione peroxide (GPX) activity was evaluated. In addition, glutathione (GSH) and malondialdehyde (MDA) the heart tissue and creatinephosphokinase-MB (CPK-MB) and troponin (TPI) levels and cholinesterase activity in the blood were evaluated. DZN-induced oxidative damage and elevated the levels of the cardiac markers CK-MB, TPI, MDA and LDH and decreased SOD, CAT and cholinesterase activity and GSH level compared with the control group. Treatment with Thy reduced DZN cardio-toxicity and cholinesterase activity. The success of Thy supplementation against DZN toxicity can be attributed to the antioxidant effects of its constituents. Administration of Thy as a natural antioxidant decreased DZN cardio-toxicity and improved cholinesterase activity in rats through the mechanism of free radical scavenging.
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Selective Vulnerability of Brainstem Nuclei in Distinct Tauopathies: A Postmortem Study.
Eser, Rana A; Ehrenberg, Alexander J; Petersen, Cathrine; Dunlop, Sara; Mejia, Maria B; Suemoto, Claudia K; Walsh, Christine M; Rajana, Hima; Oh, Jun; Theofilas, Panos; Seeley, William W; Miller, Bruce L; Neylan, Thomas C; Heinsen, Helmut; Grinberg, Lea T
2018-02-01
The brainstem nuclei of the reticular formation (RF) are critical for regulating homeostasis, behavior, and cognition. RF degenerates in tauopathies including Alzheimer disease (AD), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD). Although the burden of phopho-tau inclusion is high across these diseases, suggesting a similar vulnerability pattern, a distinct RF-associated clinical phenotype in these diseases indicates the opposite. To compare patterns of RF selective vulnerability to tauopathies, we analyzed 5 RF nuclei in tissue from 14 AD, 14 CBD, 10 PSP, and 3 control cases. Multidimensional quantitative analysis unraveled discernable differences on how these nuclei are vulnerable to AD, CBD, and PSP. For instance, PSP and CBD accrued more tau inclusions than AD in locus coeruleus, suggesting a lower vulnerability to AD. However, locus coeruleus neuronal loss in AD was so extreme that few neurons remained to develop aggregates. Likewise, tau burden in gigantocellular nucleus was low in AD and high in PSP, but few GABAergic neurons were present in AD. This challenges the hypothesis that gigantocellular nucleus neuronal loss underlies REM behavioral disorders because REM behavioral disorders rarely manifests in AD. This study provides foundation for characterizing the clinical consequences of RF degeneration in tauopathies and guiding customized treatment. © 2018 American Association of Neuropathologists, Inc. All rights reserved.
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An integrated multi-electrode-optrode array for in vitro optogenetics
Welkenhuysen, Marleen; Hoffman, Luis; Luo, Zhengxiang; De Proft, Anabel; Van den Haute, Chris; Baekelandt, Veerle; Debyser, Zeger; Gielen, Georges; Puers, Robert; Braeken, Dries
2016-01-01
Modulation of a group of cells or tissue needs to be very precise in order to exercise effective control over the cell population under investigation. Optogenetic tools have already demonstrated to be of great value in the study of neuronal circuits and in neuromodulation. Ideally, they should permit very accurate resolution, preferably down to the single cell level. Further, to address a spatially distributed sample, independently addressable multiple optical outputs should be present. In current techniques, at least one of these requirements is not fulfilled. In addition to this, it is interesting to directly monitor feedback of the modulation by electrical registration of the activity of the stimulated cells. Here, we present the fabrication and characterization of a fully integrated silicon-based multi-electrode-optrode array (MEOA) for in vitro optogenetics. We demonstrate that this device allows for artifact-free electrical recording. Moreover, the MEOA was used to reliably elicit spiking activity from ChR2-transduced neurons. Thanks to the single cell resolution stimulation capability, we could determine spatial and temporal activation patterns and spike latencies of the neuronal network. This integrated approach to multi-site combined optical stimulation and electrical recording significantly advances today’s tool set for neuroscientists in their search to unravel neuronal network dynamics. PMID:26832455
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Heldt, Scott A.; Elberger, Andrea J.; Deng, Yunping; Guley, Natalie H.; Del Mar, Nobel; Rogers, Joshua; Choi, Gy Won; Ferrell, Jessica; Rex, Tonia S.; Honig, Marcia G.; Reiner, Anton
2014-01-01
Emotional disorders are a common outcome from mild traumatic brain injury (TBI) in humans, but their pathophysiological basis is poorly understood. We have developed a mouse model of closed-head blast injury using an air pressure wave delivered to a small area on one side of the cranium, to create mild TBI. We found that 20-psi blasts in 3-month-old C57BL/6 male mice yielded no obvious behavioral or histological evidence of brain injury, while 25–40 psi blasts produced transient anxiety in an open field arena but little histological evidence of brain damage. By contrast, 50–60 psi blasts resulted in anxiety-like behavior in an open field arena that became more evident with time after blast. In additional behavioral tests conducted 2–8 weeks after blast, 50–60 psi mice also demonstrated increased acoustic startle, perseverance of learned fear, and enhanced contextual fear, as well as depression-like behavior and diminished prepulse inhibition. We found no evident cerebral pathology, but did observe scattered axonal degeneration in brain sections from 50 to 60 psi mice 3–8 weeks after blast. Thus, the TBI caused by single 50–60 psi blasts in mice exhibits the minimal neuronal loss coupled to “diffuse” axonal injury characteristic of human mild TBI. A reduction in the abundance of a subpopulation of excitatory projection neurons in basolateral amygdala enriched in Thy1 was, however, observed. The reported link of this neuronal population to fear suppression suggests their damage by mild TBI may contribute to the heightened anxiety and fearfulness observed after blast in our mice. Our overpressure air blast model of concussion in mice will enable further studies of the mechanisms underlying the diverse emotional deficits seen after mild TBI. PMID:24478749
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The comparison of acoustic and psychic parameters of subjective tinnitus.
Karatas, Erkan; Deniz, Murat
2012-02-01
We aim to assess the correlation between audiometric data, and psychotic and acoustic measures associated with subjective tinnitus (ST) and to clarify the importance of the psychological process in determining the degree of subjective annoyance and disability due to tinnitus. Fifty-four patients experiencing unilateral ST were allocated for the study. Acoustic assessment of patients including LDL (loudness discomfort levels), MML (minimum masking level) and RI (residual inhibition) was performed. Tinnitus Handicap Inventory (THI), Beck Depression Inventory (BDI) and Visual Analog Scale (VAS) tests were performed for the psychological aspects of subjective annoyance. RI was positive in 23 patients with 13 frequency-matched stimuli at 8,000 Hz. Masking treatment response was successful in 16 RI-positive patients. Mean and standard deviation (SD) of THI scores were 38.77 ± 23.63. Ten patients (%18.51) with tinnitus had ≥ 17 points score, which was significant for BDI. Mean and SD were 5.01 ± 2.31 for VAS-1 scores (severity of tinnitus), 7.98 ± 2.79 for VAS-2 (frequency and duration of tinnitus), 5.77 ± 2.72 for VAS-3 (discomfort level), 3.56 ± 3.30 for VAS-4 (attention deficit) and 3.31 ± 3.31 for VAS-5 (sleep disorders). A significant correlation was found between the tinnitus duration time, age, gender and THI scores (P < 0.05). There were statistically significant correlations between VAS 1, 2, 3 scores and LDL, MML and RI (P > 0.05). RI might be largely frequency dependent and was found as an indicator for the masking treatment response. We did not notice statistically significant correlations between audiometric data and THI and BDI. There were correlations between with VAS and LDL and with MML and RI. VAS was simpler and easier for the assessment of ST. We should consider the psychological aspects of ST and assess it as a symptom separately with acoustic and psychotic tests.
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Goscinny, Séverine; Hanot, Vincent; Trabelsi, Hasna; Van Loco, Joris
2014-01-01
2-Methylimidazole, 4-methylimidazole (2-MI and 4-MI), 2-acetyl-4-(1,2,3,4-tetrahydroxybutyl) imidazole (THI) and 5-hydroxymethylfurfural (5-HMF) are neo-formed compounds generated during the manufacture of caramel colours and are transferred to the processed food. These contaminants are known to have a toxicological profile that may pose health risks. Hence, to characterise THI, 2- and 4-MI and 5-HMF levels in liquid foods, an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and sample preparation was divided into two analytical strategies depending on the concentration range expected in the type of foods targeted. For the determination of the imidazole substitutes (THI, 2- and 4-MI), a sample enrichment and clean-up step by strong cation solid-phase extraction was developed. This method is capable of quantifying over a range of 5 ng ml⁻¹ (LOQ) to 500 ng ml⁻¹ with recoveries of 75.4-112.4% and RSDs of 1.5-15%. For determination of 5-HMF, a standard addition method was applied covering the linear range of 0.25-30 µg ml⁻¹ with RSDs from 2.8% (for intraday precision) to 9.2% (for intermediate precision). The validated analytical methods were applied to 28 liquid food samples purchased from local markets. THI was found only in the beer samples at levels up to 141.2 ng ml⁻¹. For 2-MI, non-quantifiable traces were observed for all samples, while 4-MI was observed in all samples with large concentration variations (from < LOQ to 563.9 ng ml⁻¹). 5-HMF was found at expected concentrations, except for a sherry vinegar sample (113 µg ml⁻¹), which required a high level of dilution before following the standard addition protocol.
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ODE constrained mixture modelling: a method for unraveling subpopulation structures and dynamics.
Hasenauer, Jan; Hasenauer, Christine; Hucho, Tim; Theis, Fabian J
2014-07-01
Functional cell-to-cell variability is ubiquitous in multicellular organisms as well as bacterial populations. Even genetically identical cells of the same cell type can respond differently to identical stimuli. Methods have been developed to analyse heterogeneous populations, e.g., mixture models and stochastic population models. The available methods are, however, either incapable of simultaneously analysing different experimental conditions or are computationally demanding and difficult to apply. Furthermore, they do not account for biological information available in the literature. To overcome disadvantages of existing methods, we combine mixture models and ordinary differential equation (ODE) models. The ODE models provide a mechanistic description of the underlying processes while mixture models provide an easy way to capture variability. In a simulation study, we show that the class of ODE constrained mixture models can unravel the subpopulation structure and determine the sources of cell-to-cell variability. In addition, the method provides reliable estimates for kinetic rates and subpopulation characteristics. We use ODE constrained mixture modelling to study NGF-induced Erk1/2 phosphorylation in primary sensory neurones, a process relevant in inflammatory and neuropathic pain. We propose a mechanistic pathway model for this process and reconstructed static and dynamical subpopulation characteristics across experimental conditions. We validate the model predictions experimentally, which verifies the capabilities of ODE constrained mixture models. These results illustrate that ODE constrained mixture models can reveal novel mechanistic insights and possess a high sensitivity.
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The influence of donor age on liver regeneration and hepatic progenitor cell populations.
Ono, Yoshihiro; Kawachi, Shigeyuki; Hayashida, Tetsu; Wakui, Masatoshi; Tanabe, Minoru; Itano, Osamu; Obara, Hideaki; Shinoda, Masahiro; Hibi, Taizo; Oshima, Go; Tani, Noriyuki; Mihara, Kisyo; Kitagawa, Yuko
2011-08-01
Recent reports suggest that donor age might have a major impact on recipient outcome in adult living donor liver transplantation (LDLT), but the reasons underlying this effect remain unclear. The aims of this study were to compare liver regeneration between young and aged living donors and to evaluate the number of Thy-1+ cells, which have been reported to be human hepatic progenitor cells. LDLT donors were divided into 2 groups (Group O, donor age ≥ 50 years, n = 6 and Group Y, donor age ≤ 30 years, n = 9). The remnant liver regeneration rates were calculated on the basis of computed tomography volumetry on postoperative days 7 and 30. Liver tissue samples were obtained from donors undergoing routine liver biopsy or patients undergoing partial hepatectomy for metastatic liver tumors. Thy-1+ cells were isolated and counted using immunomagnetic activated cell sorting (MACS) technique. Donor liver regeneration rates were significantly higher in young donors compared to old donors (P = .042) on postoperative day 7. Regeneration rates were significantly higher after right lobe resection compared to rates after left lobe resection. The MACS findings showed that the number of Thy-1+ cells in the human liver consistently tended to decline with age. Our study revealed that liver regeneration is impaired with age after donor hepatectomy, especially after right lobe resection. The declining hepatic progenitor cell population might be one of the reasons for impaired liver regeneration in aged donors. Copyright © 2011 Mosby, Inc. All rights reserved.
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Ryge, Jesper; Winther, Ole; Wienecke, Jacob; Sandelin, Albin; Westerdahl, Ann-Charlotte; Hultborn, Hans; Kiehn, Ole
2010-06-09
Spinal cord injury leads to neurological dysfunctions affecting the motor, sensory as well as the autonomic systems. Increased excitability of motor neurons has been implicated in injury-induced spasticity, where the reappearance of self-sustained plateau potentials in the absence of modulatory inputs from the brain correlates with the development of spasticity. Here we examine the dynamic transcriptional response of motor neurons to spinal cord injury as it evolves over time to unravel common gene expression patterns and their underlying regulatory mechanisms. For this we use a rat-tail-model with complete spinal cord transection causing injury-induced spasticity, where gene expression profiles are obtained from labeled motor neurons extracted with laser microdissection 0, 2, 7, 21 and 60 days post injury. Consensus clustering identifies 12 gene clusters with distinct time expression profiles. Analysis of these gene clusters identifies early immunological/inflammatory and late developmental responses as well as a regulation of genes relating to neuron excitability that support the development of motor neuron hyper-excitability and the reappearance of plateau potentials in the late phase of the injury response. Transcription factor motif analysis identifies differentially expressed transcription factors involved in the regulation of each gene cluster, shaping the expression of the identified biological processes and their associated genes underlying the changes in motor neuron excitability. This analysis provides important clues to the underlying mechanisms of transcriptional regulation responsible for the increased excitability observed in motor neurons in the late chronic phase of spinal cord injury suggesting alternative targets for treatment of spinal cord injury. Several transcription factors were identified as potential regulators of gene clusters containing elements related to motor neuron hyper-excitability, the manipulation of which potentially could be used to alter the transcriptional response to prevent the motor neurons from entering a state of hyper-excitability.
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Transcriptional regulation of gene expression clusters in motor neurons following spinal cord injury
2010-01-01
Background Spinal cord injury leads to neurological dysfunctions affecting the motor, sensory as well as the autonomic systems. Increased excitability of motor neurons has been implicated in injury-induced spasticity, where the reappearance of self-sustained plateau potentials in the absence of modulatory inputs from the brain correlates with the development of spasticity. Results Here we examine the dynamic transcriptional response of motor neurons to spinal cord injury as it evolves over time to unravel common gene expression patterns and their underlying regulatory mechanisms. For this we use a rat-tail-model with complete spinal cord transection causing injury-induced spasticity, where gene expression profiles are obtained from labeled motor neurons extracted with laser microdissection 0, 2, 7, 21 and 60 days post injury. Consensus clustering identifies 12 gene clusters with distinct time expression profiles. Analysis of these gene clusters identifies early immunological/inflammatory and late developmental responses as well as a regulation of genes relating to neuron excitability that support the development of motor neuron hyper-excitability and the reappearance of plateau potentials in the late phase of the injury response. Transcription factor motif analysis identifies differentially expressed transcription factors involved in the regulation of each gene cluster, shaping the expression of the identified biological processes and their associated genes underlying the changes in motor neuron excitability. Conclusions This analysis provides important clues to the underlying mechanisms of transcriptional regulation responsible for the increased excitability observed in motor neurons in the late chronic phase of spinal cord injury suggesting alternative targets for treatment of spinal cord injury. Several transcription factors were identified as potential regulators of gene clusters containing elements related to motor neuron hyper-excitability, the manipulation of which potentially could be used to alter the transcriptional response to prevent the motor neurons from entering a state of hyper-excitability. PMID:20534130
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Magrin, L; Brscic, M; Lora, I; Rumor, C; Tondello, L; Cozzi, G; Gottardo, F
2017-06-01
This research aimed at assessing the effects of a ceiling fan ventilation system on health, feeding, social behaviour and growth response of finishing young bulls fattened indoors during a mild summer season. A total of 69 Charolais young bulls were housed in six pens without any mechanical ventilation system (Control) and in six pens equipped with ceiling fans. The experimental period lasted 98 days from June until mid-September 2014. Four experimental days were considered in order to assess the effect of the ventilation system under two different microclimatic conditions: 2 alert days at monthly interval with temperature humidity index (THI) between 75 and 78, and 2 normal days with THI⩽74. Health and behaviour of the bulls were evaluated through 8-h observation sessions starting after morning feed delivery. The study was carried out during a rather cool summer with a climate average THI of 68.9 and 4 days with average THI>75. Despite these mild climate conditions, ceiling fans lowered litter moisture and acted as a preventive measure for bulls' dirtiness (odd ratio=47.9; 95% CI 19.6 to 117.4). The risk of abnormal breathing was increased for Control bulls (odd ratio=40.7; 95% CI 5.4 to 304.2). When exposed to alert THI conditions, respiration rate and panting scores increased and rumination duration dropped in Control bulls compared with bulls provided with a ceiling fan. During observations under alert THI, bulls spent less time eating, more time being inactive and consumed more water compared with normal THI conditions. Bulls' daily dry matter intake measured during the observation sessions decreased on alert compared with normal THI days (P<0.001) due to a drop of intake during the daylight hours. Ceiling fan treatment had no effect on bulls' growth performance or water consumption but these results most likely depended on the mild climate conditions. Ceiling fans proved to mitigate some of the negative effects of heat stress on bulls' behaviour (rumination, lying down and drinking water) and respiration rate, however. The lack of a significant improvement of bulls' growth response should not discourage beef farmers from using ceiling fans in indoor systems, considering the likely increase in frequency and intensity of heat waves in the planet's temperate areas induced by global warming.
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Prospective Validation of ATA and ETA Sonographic Pattern Risk of Thyroid Nodules Selected for FNAC.
Maino, Fabio; Forleo, Raffaella; Martinelli, Martina; Fralassi, Noemi; Barbato, Filomena; Pilli, Tania; Capezzone, Marco; Brilli, Lucia; Ciuoli, Cristina; Di Cairano, Giovanni; Nigi, Laura; Pacini, Furio; Castagna, Maria Grazia
2018-06-01
Recently, the American Thyroid Association (ATA) and the European Thyroid Association (ETA) have proposed that thyroid ultrasound (US) should be used to stratify the risk of malignancy in thyroid nodules and to aid decision-making about whether fine-needle aspiration cytology (FNAC) is indicated. To validate and to compare the ATA and ETA US risk stratification systems of thyroid nodules in a prospective series of thyroid nodules submitted to FNAC. We prospectively evaluated 432 thyroid nodules selected for FNAC from 340 patients. Cytology reports were based on the five categories according to the criteria of the British Thyroid Association. The proportion of Thy2 nodules decreased significantly, whereas the proportion of Thy4/Thy5 nodules significantly increased with increasing US risk class (P < 0.0001). The ability to identify benign and malignant nodules was similar between ATA and ETA systems. According to ATA and ETA US risk stratification systems, 23.7% and 56.0% nodules did not meet the criteria for FNAC, respectively. Considering only categories at lower risk of malignancy, the cumulative malignancy rate in these nodules was 1.2% for ATA and 1.7% for ETA US risk stratification systems. ETA and ATA US risk stratification systems provide effective malignancy risk stratification for thyroid nodules. In clinical practice, using this approach, we should be able to reduce the number of unnecessary FNAC without losing clinically relevant thyroid cancer.
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The effects of alprazolam on tinnitus: a cross-over randomized clinical trial.
Jalali, Mir Mohammad; Kousha, Abdorrahim; Naghavi, Sayed Ebrahim; Soleimani, Robabeh; Banan, Rozbeh
2009-11-01
Tinnitus remains a phenomenon with an unknown pathophysiology and for which few therapeutic measures are available. To date there has been insufficient evidence to support the use of alprazolam in the treatment of tinnitus. We sought to evaluate the efficacy of alprazolam for relief of tinnitus. Thirty-six tinnitus sufferers participated in this cross-over, randomized, triple-blind, placebo-controlled trial. Inclusion criteria included patients between ages 21 and 65, with a complaint of non-pulsatile tinnitus of more than 1 year duration. Patients with depressive or anxiety disorders were excluded, as were those using hearing aids. Participants received alprazolam 1.5 mg daily versus placebo in each period. Primary outcome variables included the Tinnitus Handicap Inventory (THI), a Visual Analog Scale (VAS), and tinnitus loudness. Thirty patients completed the study. The average age of patients was 47.58+/-7.65 years. Alprazolam in comparison with placebo did not result in statistically significantly greater relief in THI score and tinnitus loudness. There was a significant improvement in VAS score in the alprazolam group compared with the placebo group (p<0.001). These results suggest that although alprazolam did not improve the THI score or sensation level of loudness significantly, it has a desirable effect on VAS. Further work is needed to determine the beneficial effects of alprazolam in distressed or depressed patients.
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Kreuzer, Peter M; Poeppl, Timm B; Bulla, Jan; Schlee, Winfried; Lehner, Astrid; Langguth, Berthold; Schecklmann, Martin
2016-10-01
Interference of ongoing neuronal activity and brain stimulation motivated this study to combine repetitive transcranial magnetic stimulation (rTMS) and relaxation techniques in tinnitus patients. Forty-two patients were enrolled in this one-arm proof-of-concept study to receive ten sessions of rTMS applied to the left dorsolateral prefrontal cortex and temporo-parietal cortex. During stimulation, patients listened to five different kinds of relaxation audios. Variables of interest were tinnitus questionnaires, tinnitus numeric rating scales, depressivity, and quality of life. Results were compared to results of historical control groups having received the same rTMS protocol (active control) and sham treatment (placebo) without relaxation techniques. Thirty-eight patients completed the treatment, drop-out rates and adverse events were low. Responder rates (reduction in tinnitus questionnaire (TQ) score ≥5 points 10 weeks after treatment) were 44.7 % in the study, 27.8 % in the active control group, and 21.7 % in the placebo group, differing between groups on a near significant level. For the tinnitus handicap inventory (THI), the main effect of group was not significant. However, linear mixed model analyses showed that the relaxation/rTMS group differed significantly from the active control group showing steeper negative THI trend for the relaxation/rTMS group indicating better amelioration over the course of the trial. Deepness of relaxation during rTMS and selection of active relaxation vs. passive listening to music predicted larger TQ. All remaining secondary outcomes turned out non-significant. This combined treatment proved to be a safe, feasible and promising approach to enhance rTMS treatment effects in chronic tinnitus.
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West, J W; Mullinix, B G; Bernard, J K
2003-01-01
Lactating cows were exposed to moderate and hot, humid weather to determine the effect of increasing ambient temperature, relative humidity, or temperature-humidity index (THI) on intake, milk yield, and milk temperature. Minimum and maximum temperatures averaged 17.9 and 29.5 degrees C (cool period) and 22.5 and 34.4 degrees C (hot period), and minimum and maximum THI averaged 63.8 and 76.6 (cool period) and 72.1 and 83.6 (hot period). Environmental conditions had minor effects on intake and milk yield during the cool period. During the hot period, the THI 2 d earlier and mean air temperature 2 d earlier had the greatest impact on milk yield and DMI, respectively. Both breeds maintained milk temperature within normal ranges during the cool period, but Holstein and Jersey p.m. milk temperatures averaged 39.6 and 39.2 degrees C during the hot period. Current day mean air temperature during the hot period had the greatest impact on cow p.m. milk temperature, and minimum air temperature had the greatest influence on a.m. milk temperature. Dry matter intake and milk yield declined linearly with respective increases in air temperature or THI during the hot period and milk temperature increased linearly with increasing air temperature. Dry matter intake and milk yield both exhibited a curvilinear relationship with milk temperature. Environmental modifications should target the effects of high temperatures on cow body temperature and should modify the environment at critical times during the day when cows are stressed, including morning hours when ambient temperatures are typically cooler and cows are not assumed to be stressed.
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Uneven colonization of the lymphoid periphery by T cells that undergo early TCRα rearrangements1
Hendricks, Deborah W.; Fink, Pamela J.
2009-01-01
A sparse population of thymocytes undergoes TCRα gene rearrangement early in development, before the double positive stage. The potential of these cells to contribute to the peripheral T cell pool is unknown. To examine the peripheral T cell compartment expressing a repertoire biased to early TCR gene rearrangements, we developed a mouse model in which TCRα rearrangements are restricted to the double negative stage of thymocyte development. These mice carry floxed RAG2 alleles and a Cre transgene driven by the CD4 promoter. As expected, conventional T cell development is compromised in such Cre(+) RAG2fl/fl mice, and the TCRαβ+ T cells that develop are limited in their TCRα repertoire, preferentially utilizing early-rearranging Vα genes. In the gut, the Thy-1+TCRαβ+ intraepithelial lymphocyte (IEL) compartment is surprisingly intact, while the Thy-1−TCRαβ+ subset is almost completely absent. Thus, T cells expressing a TCRα repertoire that is the product of early gene rearrangements can preferentially populate distinct IEL compartments. Despite this capacity, Cre(+) RAG2fl/fl T cell progenitors cannot compete with wild-type (WT) T cell progenitors in mixed bone marrow chimeras, suggesting that in normal mice, there is only a small contribution to the peripheral T cell pool by cells that have undergone early TCRα rearrangements. In the absence of WT competitors, aggressive homeostatic proliferation in the IEL compartment can promote a relatively normal Thy-1+ TCRαβ+ T cell pool from the limited population derived from Cre(+) RAG2fl/fl progenitors. PMID:19299725
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Favorable prognostic role of tropomodulins in neuroblastoma.
Bettinsoli, Paola; Ferrari-Toninelli, Giulia; Bonini, Sara Anna; Guarienti, Michela; Cangelosi, Davide; Varesio, Luigi; Memo, Maurizio
2018-06-05
Neuroblastoma is a pediatric tumor of the sympatoadrenal lineage of the neural crest characterized by high molecular and clinical heterogeneity, which are the main causes of the poor response to standard multimodal therapy. The identification of new and selective biomarkers is important to improve our knowledge on the mechanisms of neuroblastoma progression and to find the targets for innovative cancer therapies. This study identifies a positive correlation among tropomodulins (TMODs) proteins expression and neuroblastoma progression. TMODs bind the pointed end of actin filaments, regulate polymerization and depolymerization processes modifying actin cytoskeletal dynamic and influencing neuronal development processes. Expression levels of TMODs genes were analyzed in 17 datasets comprising different types of tumors, including neuroblastoma, and it was demonstrated that high levels of tropomodulin1 ( TMOD1 ) and tropomodulin 2 ( TMOD2 ) correlate positively with high survival probability and with favorable clinical and molecular characteristics. Functional studies on neuroblastoma cell lines, showed that TMOD1 knockin induced cell cycle arrest, cell proliferation arrest and a mature functional differentiation. TMOD1 overexpression was responsible for particular cell morphology and biochemical changes which directed cells towards a neuronal favorable differentiation profile. TMOD1 downregulation also induced cell proliferation arrest but caused the loss of mature cell differentiation and promoted the development of neuroendocrine cellular characteristics, delineating an aggressive and unfavorable tumor behavior. Overall, these data indicated that TMODs are favorable prognostic biomarkers in neuroblastoma and we believe that they could contribute to unravel a new pathophysiological mechanism of neuroblastoma resistance contributing to the design of personalized therapeutics opportunities.
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Quintá, HR; Galigniana, MD
2012-01-01
BACKGROUND AND PURPOSE The immunosuppressive macrolide FK506 (tacrolimus) shows neuroregenerative action by a mechanism that appears to involve the Hsp90-binding immunophilin FKBP52. This study analyses some aspects of the early steps of neuronal differentiation and neuroregeneration. EXPERIMENTAL APPROACH Undifferentiated murine neuroblastoma cells and hippocampal neurones isolated from embryonic day-17 rat embryos were induced to differentiate with FK506. Subcellular relocalization of FKBP52, Hsp90 and its co-chaperone p23 was analysed by indirect immunofluorescence confocal microscopy and by Western blots of axonal fractions isolated from cells grown on a porous transwell cell culture chamber. Neuroregeneration was evaluated using a scratch-wound assay. KEY RESULTS In undifferentiated cells, FKBP52, Hsp90 and p23 are located in the cell nucleus, forming an annular structure that disassembles when the differentiation process is triggered by FK506. This was observed in the N2a cell line and in hippocampal neurones. More importantly, the annular structure of chaperones is reassembled after damaging the neurones, whereas FK506 prompts their rapid regeneration, a process linked to the subcellular redistribution of the heterocomplex. CONCLUSIONS AND IMPLICATIONS There is a direct relationship between the disassembly of the chaperone complex and the progression of neuronal differentiation upon stimulation with the immunophilin ligand FK506. Both neuronal differentiation and neuroregeneration appear to be mechanistically linked, so the elucidation of one mechanism may lead to unravel the properties of the other. This study also implies that the discovery of FK506 derivatives, devoid of immunosuppressive action, would be therapeutically significant for neurotrophic use. PMID:22091865
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Nassi, Jonathan J.; Cepko, Constance L.; Born, Richard T.; Beier, Kevin T.
2015-01-01
The nervous system is complex not simply because of the enormous number of neurons it contains but by virtue of the specificity with which they are connected. Unraveling this specificity is the task of neuroanatomy. In this endeavor, neuroanatomists have traditionally exploited an impressive array of tools ranging from the Golgi method to electron microscopy. An ideal method for studying anatomy would label neurons that are interconnected, and, in addition, allow expression of foreign genes in these neurons. Fortuitously, nature has already partially developed such a method in the form of neurotropic viruses, which have evolved to deliver their genetic material between synaptically connected neurons while largely eluding glia and the immune system. While these characteristics make some of these viruses a threat to human health, simple modifications allow them to be used in controlled experimental settings, thus enabling neuroanatomists to trace multi-synaptic connections within and across brain regions. Wild-type neurotropic viruses, such as rabies and alpha-herpes virus, have already contributed greatly to our understanding of brain connectivity, and modern molecular techniques have enabled the construction of recombinant forms of these and other viruses. These newly engineered reagents are particularly useful, as they can target genetically defined populations of neurons, spread only one synapse to either inputs or outputs, and carry instructions by which the targeted neurons can be made to express exogenous proteins, such as calcium sensors or light-sensitive ion channels, that can be used to study neuronal function. In this review, we address these uniquely powerful features of the viruses already in the neuroanatomist’s toolbox, as well as the aspects of their biology that currently limit their utility. Based on the latter, we consider strategies for improving viral tracing methods by reducing toxicity, improving control of transsynaptic spread, and extending the range of species that can be studied. PMID:26190977
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Lymphocyte functions in dairy cows in hot environment
NASA Astrophysics Data System (ADS)
Lacetera, Nicola; Bernabucci, Umberto; Scalia, Daniela; Ronchi, Bruno; Kuzminsky, Giorgina; Nardone, Alessandro
2005-11-01
This study was carried out to ascertain the effects of intense high environmental temperatures (HET) on lymphocyte functions in periparturient dairy cows. The study was undertaken from the beginning of March through the end of July 2003 in a commercial dairy unit located approximately 40 km north of Rome. Thirty-four Holstein cows were utilised in the study. Twenty-two of these cows gave birth in spring (SP cows), from 28 March to 30 April. The remaining 12 cows gave birth in summer (SU cows), between 15 June and 2 July. The two groups of cows were balanced for parity and were fed the same rations. Blood samples were taken 4, 3, 2 and 1 week before calving, and 1, 2 and 4 weeks after calving, in order to evaluate peripheral blood mononuclear cell (PBMC) function in vitro, and to determine plasma cortisol concentrations. After isolation, the PBMC were stimulated with mitogens and their response in terms of DNA synthesis and IgM secretion was measured. During spring, either the day (9 20 h) or the night (21 8 h) temperature humidity index (THI) was below the upper critical THI (72) established for dairy cows. During summer, the mean daily THI values were 79.5±2.9 during the day and 70.1±4.7 during the night. Furthermore, during summer, three heat waves (a period of at least 3 consecutive days during which there were less than 10 recovery hours) occurred. Recovery hours were intended hours with a THI below 72. The first heat wave lasted 5 days, the second 6 days, and the third 15 days. Compared to the SP cows, over the entire periparturient period the extent of DNA synthesis and IgM secretion levels were lower (P ranging from <0.01 to 0.0001) and higher (P<0.01) respectively, in the SU cows. Before calving, the SU cows also presented higher (P<0.01) concentrations of plasma cortisol compared to the SP cows. This study indicates that the effects of HET on the immune response depend on the specific immune function under consideration, and that neuroendocrinal changes due to HET may play a role in the perturbation of immune functions.
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Mechanisms of neuroimmune gene induction in alcoholism.
Crews, Fulton T; Vetreno, Ryan P
2016-05-01
Alcoholism is a primary, chronic relapsing disease of brain reward, motivation, memory, and related circuitry. It is characterized by an individual's continued drinking despite negative consequences related to alcohol use, which is exemplified by alcohol use leading to clinically significant impairment or distress. Chronic alcohol consumption increases the expression of innate immune signaling molecules (ISMs) in the brain that alter cognitive processes and promote alcohol drinking. Unraveling the mechanisms of alcohol-induced neuroimmune gene induction is complicated by positive loops of multiple cytokines and other signaling molecules that converge on nuclear factor kappa-light-chain-enhancer of activated B cells and activator protein-1 leading to induction of additional neuroimmune signaling molecules that amplify and expand the expression of ISMs. Studies from our laboratory employing reverse transcription polymerase chain reaction (RT-PCR) to assess mRNA, immunohistochemistry and Western blot analysis to assess protein expression, and others suggest that ethanol increases brain neuroimmune gene and protein expression through two distinct mechanisms involving (1) systemic induction of innate immune molecules that are transported from blood to the brain and (2) the direct release of high-mobility group box 1 (HMGB1) from neurons in the brain. Released HMGB1 signals through multiple receptors, particularly Toll-like receptor (TLR) 4, that potentiate cytokine receptor responses leading to a hyperexcitable state that disrupts neuronal networks and increases excitotoxic neuronal death. Innate immune gene activation in brain is persistent, consistent with the chronic relapsing disease that is alcoholism. Expression of HMGB1, TLRs, and other ISMs is increased several-fold in the human orbital frontal cortex, and expression of these molecules is highly correlated with each other as well as lifetime alcohol consumption and age of drinking onset. The persistent and cumulative nature of alcohol on HMGB1 and TLR gene induction support their involvement in alcohol-induced long-term changes in brain function and neurodegeneration.
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Asano, Masanao; Li, Yue-Sheng; Núñez, Gabriel
2017-01-01
Although B cell development requires expression of the B cell antigen receptor (BCR), it remains unclear whether engagement of self-antigen provides a positive impact for most B cells. Here, we show that BCR engagement by self-ligand during development in vivo results in up-regulation of the Nod-like receptor member Nod1, which recognizes the products of intestinal commensal bacteria. In anti-thymocyte/Thy-1 autoreactive BCR knock-in mice lacking self–Thy-1 ligand, immunoglobulin light chain editing occurred, generating B cells with up-regulated Nod1, including follicular and marginal zone B cells with natural autoreactivity. This BCR editing with increased Nod1 resulted in preferential survival. In normal adult mice, most mature B cells are enriched for Nod1 up-regulated cells, and signaling through Nod1 promotes competitive survival of mature B cells. These findings demonstrate a role for microbial products in promoting survival of mature B cells through up-regulated Nod1, providing a positive effect of BCR engagement on development of most B cells. PMID:28878001
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Potent activity of the HIV-1 maturation inhibitor bevirimat in SCID-hu Thy/Liv mice.
Stoddart, Cheryl A; Joshi, Pheroze; Sloan, Barbara; Bare, Jennifer C; Smith, Philip C; Allaway, Graham P; Wild, Carl T; Martin, David E
2007-11-28
The HIV-1 maturation inhibitor, 3-O-(3',3'-dimethylsuccinyl) betulinic acid (bevirimat, PA-457) is a promising drug candidate with 10 nM in vitro antiviral activity against multiple wild-type (WT) and drug-resistant HIV-1 isolates. Bevirimat has a novel mechanism of action, specifically inhibiting cleavage of spacer peptide 1 (SP1) from the C-terminus of capsid which results in defective core condensation. Oral administration of bevirimat to HIV-1-infected SCID-hu Thy/Liv mice reduced viral RNA by >2 log(10) and protected immature and mature T cells from virus-mediated depletion. This activity was observed at plasma concentrations that are achievable in humans after oral dosing, and bevirimat was active up to 3 days after inoculation with both WT HIV-1 and an AZT-resistant HIV-1 clinical isolate. Consistent with its mechanism of action, bevirimat caused a dose-dependent inhibition of capsid-SP1 cleavage in HIV-1-infected human thymocytes obtained from these mice. HIV-1 NL4-3 with an alanine-to-valine substitution at the N-terminus of SP1 (SP1/A1V), which is resistant to bevirimat in vitro, was also resistant to bevirimat treatment in the mice, and SP1/AIV had replication and thymocyte kinetics similar to that of WT NL4-3 with no evidence of fitness impairment in in vivo competition assays. Interestingly, protease inhibitor-resistant HIV-1 with impaired capsid-SP1 cleavage was hypersensitive to bevirimat in vitro with a 50% inhibitory concentration 140 times lower than for WT HIV-1. These results support further clinical development of this first-in-class maturation inhibitor and confirm the usefulness of the SCID-hu Thy/Liv model for evaluation of in vivo antiretroviral efficacy, drug resistance, and viral fitness.
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Strogatz, S H
2001-03-08
The study of networks pervades all of science, from neurobiology to statistical physics. The most basic issues are structural: how does one characterize the wiring diagram of a food web or the Internet or the metabolic network of the bacterium Escherichia coli? Are there any unifying principles underlying their topology? From the perspective of nonlinear dynamics, we would also like to understand how an enormous network of interacting dynamical systems-be they neurons, power stations or lasers-will behave collectively, given their individual dynamics and coupling architecture. Researchers are only now beginning to unravel the structure and dynamics of complex networks.
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Effect of heat stress on reproductive performances of dairy cattle and buffaloes: A review
Dash, Soumya; Chakravarty, A. K.; Singh, Avtar; Upadhyay, Arpan; Singh, Manvendra; Yousuf, Saleem
2016-01-01
Heat stress has adverse effects on the reproductive performances of dairy cattle and buffaloes. The dairy sector is a more vulnerable to global warming and climate change. The temperature humidity index (THI) is the widely used index to measure the magnitude of heat stress in animals. The objective of this paper was to assess the decline in performances of reproductive traits such as service period, conception rate and pregnancy rate of dairy cattle and buffaloes with respect to increase in THI. The review stated that service period in cattle is affected by season of calving for which cows calved in summer had the longest service period. The conception rate and pregnancy rate in dairy cattle were found decreased above THI 72 while a significant decline in reproductive performances of buffaloes was observed above threshold THI 75. The non-heat stress zone (HSZ) (October to March) is favorable for optimum reproductive performance, while fertility is depressed in HSZ (April to September) and critical HSZ (CHSZ) (May and June). Heat stress in animals has been associated with reduced fertility through its deleterious impact on oocyte maturation and early embryo development. The management strategies viz., nutrition modification, environment modification and timed artificial insemination protocol are to be strictly operated to ameliorate the adverse effects of heat stress in cattle and buffaloes during CHSZ to improve their fertility. The identification of genes associated with heat tolerance, its incorporation into breeding program and the inclusion of THI covariate effects in selection index should be targeted for genetic evaluation of dairy animals in the hot climate. PMID:27057105
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Effect of heat stress on reproductive performances of dairy cattle and buffaloes: A review.
Dash, Soumya; Chakravarty, A K; Singh, Avtar; Upadhyay, Arpan; Singh, Manvendra; Yousuf, Saleem
2016-03-01
Heat stress has adverse effects on the reproductive performances of dairy cattle and buffaloes. The dairy sector is a more vulnerable to global warming and climate change. The temperature humidity index (THI) is the widely used index to measure the magnitude of heat stress in animals. The objective of this paper was to assess the decline in performances of reproductive traits such as service period, conception rate and pregnancy rate of dairy cattle and buffaloes with respect to increase in THI. The review stated that service period in cattle is affected by season of calving for which cows calved in summer had the longest service period. The conception rate and pregnancy rate in dairy cattle were found decreased above THI 72 while a significant decline in reproductive performances of buffaloes was observed above threshold THI 75. The non-heat stress zone (HSZ) (October to March) is favorable for optimum reproductive performance, while fertility is depressed in HSZ (April to September) and critical HSZ (CHSZ) (May and June). Heat stress in animals has been associated with reduced fertility through its deleterious impact on oocyte maturation and early embryo development. The management strategies viz., nutrition modification, environment modification and timed artificial insemination protocol are to be strictly operated to ameliorate the adverse effects of heat stress in cattle and buffaloes during CHSZ to improve their fertility. The identification of genes associated with heat tolerance, its incorporation into breeding program and the inclusion of THI covariate effects in selection index should be targeted for genetic evaluation of dairy animals in the hot climate.
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NASA Astrophysics Data System (ADS)
Kollmann, Tobias R.; Pettoello-Mantovani, Massimo; Katopodis, Nikos F.; Hachamovitch, Moshe; Rubinstein, Arye; Kim, Ana; Goldstein, Harris
1996-04-01
To improve the usefulness of in vivo models for the investigation of the pathophysiology of human immunodeficiency virus (HIV) infection, we modified the construction of SCID mice implanted with human fetal thymus and liver (thy/liv-SCID-hu mice) so that the peripheral blood of the mice contained significant numbers of human monocytes and T cells. After inoculation with HIV-159, a primary patient isolate capable of infecting monocytes and T cells, the modified thy/liv-SCID-hu mice developed disseminated HIV infection that was associated with plasma viremia. The development of plasma viremia and HIV infection in thy/liv-SCID-hu mice inoculated with HIV-159 was inhibited by acute treatment with human interleukin (IL) 10 but not with human IL-12. The human peripheral blood mononuclear cells in these modified thy/liv-SCID-hu mice were responsive in vivo to treatment with exogenous cytokines. Human interferon γ expression in the circulating human peripheral blood mononuclear cells was induced by treatment with IL-12 and inhibited by treatment with IL-10. Thus, these modified thy/liv-SCID-hu mice should prove to be a valuable in vivo model for examining the role of immunomodulatory therapy in modifying HIV infection. Furthermore, our demonstration of the in vivo inhibitory effect of IL-10 on acute HIV infection suggests that further studies may be warranted to evaluate whether there is a role for IL-10 therapy in preventing HIV infection in individuals soon after exposure to HIV such as for children born to HIV-infected mothers.
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40 CFR 721.600 - 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name).
Code of Federal Regulations, 2010 CFR
2010-07-01
... 40 Protection of Environment 30 2010-07-01 2010-07-01 false 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name). 721.600 Section 721.600 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL ACT SIGNIFICANT NEW USES OF CHEMICAL SUBSTANCES Significant New Uses for Specific Chemical Substances § 72...
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KNOW THY TEAM, KNOW THY ENEMY: USING DECEPTION TEAMS TO ACHIEVE AIR SUPERIORITY
2016-06-01
sea/25810495. 5 Sun Tzu , The Art of War, ed. Dallas Galvin, tran. Lionel Giles (New York: Fine Creative Media, Inc., 2003), 9. 6 Dr. Eric C...the G-2 3D U.S. Army. April 1991. Tzu , Sun . The Art of War. Edited by Dallas Galvin. Translated by Lionel Giles. New York: Fine Creative...ENDNOTES ......................................................................................................................39 BIBLIOGRAPHY
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Teenagers educating teenagers about reproductive health and their rights to confidential care.
Yanda, K
2000-01-01
This paper focuses on the efforts of the Teen Health Initiative (THI) to meet the needs of teenagers for an accurate understanding of their rights to health care in New York. In particular, THI makes the state's laws understandable and explains the legal rights of minors to health care. In addition to the extensive training for professionals who work with adolescents, THI runs a peer education program. The program provides teenagers the opportunity to discuss their rights to confidential health care and gives them the tools to present that information to other adolescents around the state. An important aspect of the THI program is that it focuses on teenagers educating teenagers. Its workshop covers areas of health care to which minors can give informed consent and that they can receive confidentiality such as mental health care, drug and alcohol counseling, as well as areas of reproductive health such as birth control, pregnancy testing, prenatal care and counseling, testing and treatment for sexually transmitted diseases, HIV/AIDS testing and treatment, and abortion. The group believes that, when fully educated and treated respectfully, most teenagers are willing and able to make responsible choices about their health and their lives.
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Code of Federal Regulations, 2011 CFR
2011-10-01
... 45 Public Welfare 3 2011-10-01 2011-10-01 false Purpose. 611.1 Section 611.1 Public Welfare Regulations Relating to Public Welfare (Continued) NATIONAL SCIENCE FOUNDATION NONDISCRIMINATION IN FEDERALLY-ASSISTED PROGRAMS OF THE NATIONAL SCIENCE FOUNDATION-EFFECTUATION OF TITLE VI OF THE CIVIL RIGHTS ACT OF 1964 § 611.1 Purpose. The purpose of thi...
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Iijima, Hisaka; Suzuki, Shosuke; Koyama, Hiroshi; Nakazawa, Minato; Wakimoto, Yuji
2018-05-01
Objective: This study investigated the relationship between occupations and health status to obtain an overall understanding of a cohort of Japanese middle-aged women, including unemployed women, who comprised approximately 30% of the sample. Participants and Methods: Participants of this study were 4,454 women aged 40-69 years, classified into the following five groups based on their occupation: unemployed, 1,432; agriculture, 439; self-employed, 1,596; white collared, 793; and blue collared, 194. Participants' perceived health was assessed using a symptoms checklist called the Todai Health Index (THI, later renamed as the Total Health Index) in a baseline survey conducted in 1993. The mortality risk of the participants was assessed using the Cox's Proportional Hazards Model. Results: The means of the percentile values on the Total Scale 1 in the THI were as follows: agriculture, 43.7; unemployed, 50.8; self-employed, 52.5; white collared, 53.0; and blue collared, 56.1, with lower percentile values indicating better perceived health. The results showed that women engaged in agriculture were in significantly better health than were those in the other four occupations. The hazard ratios (HRs) and 95% confidence intervals of the occupational groups adjusted for age, area of residence, and Total Scale 1 scores were as follows: agriculture (reference group), 1; white collared, 1.16 (0.77-1.74); self-employed, 1.25 (0.87-1.78); unemployed, 1.27 (0.91-1.77); and blue collared, 1.50 (0.86-2.60). Conclusions: Women engaged in agriculture had a significantly higher tendency to have a better health status on the THI as compared to those from the other four occupational groups, and they exhibited the lowest HR as compared to their counterparts, though not statistically significant. We concluded that the perceived health status of unemployed women was similar to that of women engaged in agriculture.
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Short communication: Effect of heat stress on nonreturn rate of Italian Holstein cows.
Biffani, S; Bernabucci, U; Vitali, A; Lacetera, N; Nardone, A
2016-07-01
The data set consisted of 1,016,856 inseminations of 191,012 first, second, and third parity Holstein cows from 484 farms. Data were collected from year 2001 through 2007 and included meteorological data from 35 weather stations. Nonreturn rate at 56 d after first insemination (NR56) was considered. A logit model was used to estimate the effect of temperature-humidity index (THI) on reproduction across parities. Then, least squares means were used to detect the THI breakpoints using a 2-phase linear regression procedure. Finally, a multiple-trait threshold model was used to estimate variance components for NR56 in first and second parity cows. A dummy regression variable (t) was used to estimate NR56 decline due to heat stress. The NR56, both for first and second parity cows, was significantly (unfavorable) affected by THI from 4 d before 5 d after the insemination date. Additive genetic variances for NR56 increased from first to second parity both for general and heat stress effect. Genetic correlations between general and heat stress effects were -0.31 for first parity and -0.45 for second parity cows. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
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Genetic effects of heat stress on milk yield of Thai Holstein crossbreds.
Boonkum, W; Misztal, I; Duangjinda, M; Pattarajinda, V; Tumwasorn, S; Sanpote, J
2011-01-01
The threshold for heat stress on milk yield of Holstein crossbreds under climatic conditions in Thailand was investigated, and genetic effects of heat stress on milk yield were estimated. Data included 400,738 test-day milk yield records for the first 3 parities from 25,609 Thai crossbred Holsteins between 1990 and 2008. Mean test-day milk yield ranged from 12.6 kg for cows with <87.5% Holstein genetics to 14.4 kg for cows with ≥93.7% Holstein genetics. Daily temperature and humidity data from 26 provincial weather stations were used to calculate a temperature-humidity index (THI). Test-day milk yield varied little with THI for first parity except above a THI of 82 for cows with ≥93.7% Holstein genetics. For third parity, test-day milk yield started to decline after a THI of 74 for cows with ≥87.5% Holstein genetics and declined more rapidly after a THI of 82. A repeatability test-day model with parities as correlated traits was used to estimate heat stress parameters; fixed effects included herd-test month-test year and breed groups, days in milk, calving age, and parity; random effects included 2 additive genetic effects, regular and heat stress, and 2 permanent environment, regular and heat stress. The threshold for effect of heat stress on test-day milk yield was set to a THI of 80. All variance component estimates increased with parity; the largest increases were found for effects associated with heat stress. In particular, genetic variance associated with heat stress quadrupled from first to third parity, whereas permanent environmental variance only doubled. However, permanent environmental variance for heat stress was at least 10 times larger than genetic variance. Genetic correlations among parities for additive effects without heat stress considered ranged from 0.88 to 0.96. Genetic correlations among parities for additive effects of heat stress ranged from 0.08 to 0.22, and genetic correlations between effects regular and heat stress effects ranged from -0.21 to -0.33 for individual parities. Effect of heat stress on Thai Holstein crossbreds increased greatly with parity and was especially large after a THI of 80 for cows with a high percentage of Holstein genetics (≥93.7%). Individual sensitivity to heat stress was more environmental than genetic for Thai Holstein crossbreds. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
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[Application of health questionnaires for health management in small- and medium-sized enterprises].
Kishida, K; Saito, M; Hasegawa, T; Aoki, S; Suzuki, S
1986-01-01
Two kinds of health questionnaires, the Todai Health Index (THI) and Cumulative Fatigue Index (CFI), were applied as a screening device for health management of workers belonging to small-medium sized enterprises. A total of 495 workers composed of 452 male workers of a glass-bottle manufacturing factory and 43 male workers of a soft-drink bottling factory were the subjects of the present study. It was found that the two kinds of health questionnaires were different from each other and have their own characteristics. Twelve scales of THI were grouped into two, the first consisting of ten scales (SUSY, RESP, EYSK, MOUT, DIGE, IMPU, MENT, DEPR, NERV, and LIFE) and the second consisting of two scales (AGGR and LISC). Nine categories of CFI were grouped into one by using principal factor analysis. It was confirmed that the twelve scale scores of THI obtained at small-medium sized factories differed from those scale scores of a reference group investigated at a large-sized enterprise. It is on the basis of the scales of aggressiveness and lies and also of the scale of mental unstability which characterizes workers, locality, job (clerical or field work), and size of industry (large or small sized) that the difference could be evaluated. Urban life characterized by a life style of staying up late at night and waking up late in the morning has been reflected on the scale of life irregularity. Irregularity of life induced by transformation of working schedule, such as two or three shifts of work and overtime, was also reflected on this scale. Two scales of THI test, i.e., many subjective symptoms and digestive organ complaints, seemed to be the representative scales indicating a close relation between work load and health level. The discriminant score for diagnosis of psychosomatic diseases is considered to be one of the most useful assessments of the individual's health condition. As mentioned above, THI is recommended as a convenient assessment method for health management of workers and for screening individuals or groups requiring health management from the total respondents belonging to small-medium sized enterprises where health administrators or professionals for health services are not available. A combined use of THI and CFI is more effective in evaluating health status of field workers than the independent use of one of these two tests, because the causal relationship between work load and health status cannot be satisfactorily observed, only through THI.
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Sleep Dysfunction and EEG Alterations in Mice Overexpressing Alpha-Synuclein
McDowell, Kimberly A.; Shin, David; Roos, Kenneth P.; Chesselet, Marie-Françoise
2018-01-01
Background: Sleep disruptions occur early and frequently in Parkinson’s disease (PD). PD patients also show a slowing of resting state activity. Alpha-synuclein is causally linked to PD and accumulates in sleep-related brain regions. While sleep problems occur in over 75% of PD patients and severely impact the quality of life of patients and caregivers, their study is limited by a paucity of adequate animal models. Objective: The objective of this study was to determine whether overexpression of wildtype alpha-synuclein could lead to alterations in sleep patterns reminiscent of those observed in PD by measuring sleep/wake activity with rigorous quantitative methods in a well-characterized genetic mouse model. Methods: At 10 months of age, mice expressing human wildtype alpha-synuclein under the Thy-1 promoter (Thy1-aSyn) and wildtype littermates underwent the subcutaneous implantation of a telemetry device (Data Sciences International) for the recording of electromyograms (EMG) and electroencephalograms (EEG) in freely moving animals. Surgeries and data collection were performed without knowledge of mouse genotype. Results: Thy1-aSyn mice showed increased non-rapid eye movement sleep during their quiescent phase, increased active wake during their active phase, and decreased rapid eye movement sleep over a 24-h period, as well as a shift in the density of their EEG power spectra toward lower frequencies with a significant decrease in gamma power during wakefulness. Conclusions: Alpha-synuclein overexpression in mice produces sleep disruptions and altered oscillatory EEG activity reminiscent of PD, and this model provides a novel platform to assess mechanisms and therapeutic strategies for sleep dysfunction in PD. PMID:24867919
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Khedr, E M; Abo-Elfetoh, N; Rothwell, J C; El-Atar, A; Sayed, E; Khalifa, H
2010-07-01
Repetitive transcranial magnetic stimulation (rTMS) applied over left temporoparietal cortex has been reported to have a long-term therapeutic effect on tinnitus. We compare the impact of 1 and 25 Hz rTMS delivered either contralateral or ipsilateral to symptoms in 62 patients with unilateral chronic tinnitus. Patients were randomly assigned to one of four treatment groups: with stimulation at 1 or 25 Hz applied either ipsilateral or contralateral to symptoms. Two thousand pulses per session were given daily for 2 weeks. Changes in tinnitus handicap inventory (THI), self-rating scores of loudness, awareness, and annoyance were measured monthly for 10 months. Duration of residual inhibition (RI) and psychiatric morbidity were evaluated monthly for 3 months. There was a significant main effect of time (P < 0.0001) and a significant time x side interaction (P = 0.032) between groups. This was because of the fact that contralateral stimulation had a greater effect on THI than ipsilateral stimulation; it was also superior to left side stimulation (P = 0.027). Ratings of loudness improved more after contralateral rTMS (P = 0.037). Twenty patients had no remaining tinnitus after 3 months; the remainder had a significant increase in RI. Patients with the shortest history of tinnitus tended to respond better to rTMS. There was a significant correlation between changes in THI score and changes in Hamilton anxiety and depression scores. Ten daily treatments of 1 and 25 Hz rTMS contralateral to the side of tinnitus have a greater beneficial effect on symptoms than either ipsilateral or left side stimulation.
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Hendricks, Deborah W; Fink, Pamela J
2009-04-01
A sparse population of thymocytes undergoes TCRalpha gene rearrangement early in development, before the double-positive stage. The potential of these cells to contribute to the peripheral T cell pool is unknown. To examine the peripheral T cell compartment expressing a repertoire biased to early TCR gene rearrangements, we developed a mouse model in which TCRalpha rearrangements are restricted to the double-negative stage of thymocyte development. These mice carry floxed RAG2 alleles and a Cre transgene driven by the CD4 promoter. As expected, conventional T cell development is compromised in such Cre(+) RAG2(fl/fl) mice, and the TCRalphabeta(+) T cells that develop are limited in their TCRalpha repertoire, preferentially using early rearranging Valpha genes. In the gut, the Thy-1(+)TCRalphabeta(+) intraepithelial lymphocyte (IEL) compartment is surprisingly intact, whereas the Thy-1(-)TCRalphabeta(+) subset is almost completely absent. Thus, T cells expressing a TCRalpha repertoire that is the product of early gene rearrangements can preferentially populate distinct IEL compartments. Despite this capacity, Cre(+) RAG2(fl/fl) T cell progenitors cannot compete with wild-type T cell progenitors in mixed bone marrow chimeras, suggesting that in normal mice, there is only a small contribution to the peripheral T cell pool by cells that have undergone early TCRalpha rearrangements. In the absence of wild-type competitors, aggressive homeostatic proliferation in the IEL compartment can promote a relatively normal Thy-1(+) TCRalphabeta(+) T cell pool from the limited population derived from Cre(+) RAG2(fl/fl) progenitors.
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The ubiquitin proteasome system in glia and its role in neurodegenerative diseases
Jansen, Anne H. P.; Reits, Eric A. J.; Hol, Elly M.
2014-01-01
The ubiquitin proteasome system (UPS) is crucial for intracellular protein homeostasis and for degradation of aberrant and damaged proteins. The accumulation of ubiquitinated proteins is a hallmark of many neurodegenerative diseases, including amyotrophic lateral sclerosis, Alzheimer’s, Parkinson’s, and Huntington’s disease, leading to the hypothesis that proteasomal impairment is contributing to these diseases. So far, most research related to the UPS in neurodegenerative diseases has been focused on neurons, while glial cells have been largely disregarded in this respect. However, glial cells are essential for proper neuronal function and adopt a reactive phenotype in neurodegenerative diseases, thereby contributing to an inflammatory response. This process is called reactive gliosis, which in turn affects UPS function in glial cells. In many neurodegenerative diseases, mostly neurons show accumulation and aggregation of ubiquitinated proteins, suggesting that glial cells may be better equipped to maintain proper protein homeostasis. During an inflammatory reaction, the immunoproteasome is induced in glia, which may contribute to a more efficient degradation of disease-related proteins. Here we review the role of the UPS in glial cells in various neurodegenerative diseases, and we discuss how studying glial cell function might provide essential information in unraveling mechanisms of neurodegenerative diseases. PMID:25152710
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Identification of an aggression-promoting pheromone and its receptor neurons in Drosophila.
Wang, Liming; Anderson, David J
2010-01-14
Aggression is regulated by pheromones in many animal species. However, in no system have aggression pheromones, their cognate receptors and corresponding sensory neurons been identified. Here we show that 11-cis-vaccenyl acetate (cVA), a male-specific volatile pheromone, robustly promotes male-male aggression in the vinegar fly Drosophila melanogaster. The aggression-promoting effect of synthetic cVA requires olfactory sensory neurons (OSNs) expressing the receptor Or67d, as well as the receptor itself. Activation of Or67d-expressing OSNs, either by genetic manipulation of their excitability or by exposure to male pheromones in the absence of other classes of OSNs, is sufficient to promote aggression. High densities of male flies can promote aggression by the release of volatile cVA. In turn, cVA-promoted aggression can promote male fly dispersal from a food resource, in a manner dependent on Or67d-expressing OSNs. These data indicate that cVA may mediate negative-feedback control of male population density, through its effect on aggression. Identification of a pheromone-OSN pair controlling aggression in a genetic organism opens the way to unravelling the neurobiology of this evolutionarily conserved behaviour.
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Linking F-box protein 7 and parkin to neuronal degeneration in Parkinson's disease (PD).
Zhou, Zhi Dong; Sathiyamoorthy, Sushmitha; Angeles, Dario C; Tan, Eng King
2016-04-18
Mutations of F-box protein 7 (FBXO7) and Parkin, two proteins in ubiquitin-proteasome system (UPS), are both implicated in pathogenesis of dopamine (DA) neuron degeneration in Parkinson's disease (PD). Parkin is a HECT/RING hybrid ligase that physically receives ubiquitin on its catalytic centre and passes ubiquitin onto its substrates, whereas FBXO7 is an adaptor protein in Skp-Cullin-F-box (SCF) SCF(FBXO7) ubiquitin E3 ligase complex to recognize substrates and mediate substrates ubiquitination by SCF(FBXO7) E3 ligase. Here, we discuss the overlapping pathophysiologic mechanisms and clinical features linking Parkin and FBXO7 with autosomal recessive PD. Both proteins play an important role in neuroprotective mitophagy to clear away impaired mitochondria. Parkin can be recruited to impaired mitochondria whereas cellular stress can promote FBXO7 mitochondrial translocation. PD-linked FBXO7 can recruit Parkin into damaged mitochondria and facilitate its aggregation. WT FBXO7, but not PD-linked FBXO7 mutants can rescue DA neuron degeneration in Parkin null Drosophila. A better understanding of the common pathophysiologic mechanisms of these two proteins could unravel specific pathways for targeted therapy in PD.
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Jeong, Hee-Won; Choi, Seung-Won; Youm, Jin-Young; Lim, Jeong-Wook; Kwon, Hyon-Jo; Song, Shi-Hun
2017-11-01
Among pediatric injury, brain injury is a leading cause of death and disability. To improve outcomes, many developed countries built neurotrauma databank (NTDB) system but there was not established nationwide coverage NTDB until 2009 and there have been few studies on pediatric traumatic head injury (THI) patients in Korea. Therefore, we analyzed epidemiology and outcome from the big data of pediatric THI. We collected data on pediatric patients from 23 university hospitals including 9 regional trauma centers from 2010 to 2014 and analyzed their clinical factors (sex, age, initial Glasgow coma scale, cause and mechanism of head injury, presence of surgery). Among all the 2617 THI patients, total number of pediatric patients was 256. The average age of the subjects was 9.07 (standard deviation±6.3) years old. The male-to female ratio was 1.87 to 1 and male dominance increases with age. The most common cause for trauma were falls and traffic accidents. Age ( p =0.007), surgery ( p <0.001), mechanism of trauma ( p =0.016), subdural hemorrhage (SDH) ( p <0.001), diffuse axonal injury (DAI) ( p <0.001) were statistically significant associated with severe brain injury. Falls were the most common cause of trauma, and age, surgery, mechanism of trauma, SDH, DAI increased with injury severity. There is a critical need for effective fall and traffic accidents prevention strategies for children, and we should give attention to these predicting factors for more effective care.
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Gago-Lopez, Nuria; Awaji, Obinna; Zhang, Yiqiang; Ko, Christopher; Nsair, Ali; Liem, David; Stempien-Otero, April; MacLellan, W. Robb
2014-01-01
Summary Despite over a decade of intense research, the identity and differentiation potential of human adult cardiac progenitor cells (aCPC) remains controversial. Cardiospheres have been proposed as a means to expand aCPCs in vitro, but the identity of the progenitor cell within these 3D structures is unknown. We show that clones derived from cardiospheres could be subdivided based on expression of thymocyte differentiation antigen 1 (THY-1/CD90) into two distinct populations that exhibit divergent cardiac differentiation potential. One population, which is CD90+, expressed markers consistent with a mesenchymal/myofibroblast cell. The second clone type was CD90− and could form mature, functional myocytes with sarcomeres albeit at a very low rate. These two populations of cardiogenic clones displayed distinct cell surface markers and unique transcriptomes. Our study suggests that a rare aCPC exists in cardiospheres along with a mesenchymal/myofibroblast cell, which demonstrates incomplete cardiac myocyte differentiation. PMID:24936447
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Thatcher, Louise F.; Powell, Jonathan J.; Aitken, Elizabeth A.B.; Kazan, Kemal; Manners, John M.
2012-01-01
The LATERAL ORGAN BOUNDARIES (LOB) DOMAIN (LBD) gene family encodes plant-specific transcriptional regulators functioning in organ development. In a screen of Arabidopsis (Arabidopsis thaliana) sequence-indexed transferred DNA insertion mutants, we found disruption of the LOB DOMAIN-CONTAINING PROTEIN20 (LBD20) gene led to increased resistance to the root-infecting vascular wilt pathogen Fusarium oxysporum. In wild-type plants, LBD20 transcripts were barely detectable in leaves but abundant in roots, where they were further induced after F. oxysporum inoculation or methyl jasmonate treatment. Induction of LBD20 expression in roots was abolished in coronatine insensitive1 (coi1) and myc2 (allelic to jasmonate insensitive1) mutants, suggesting LBD20 may function in jasmonate (JA) signaling. Consistent with this, expression of the JA-regulated THIONIN2.1 (Thi2.1) and VEGETATIVE STORAGE PROTEIN2 (VSP2) genes were up-regulated in shoots of lbd20 following treatment of roots with F. oxysporum or methyl jasmonate. However, PLANT DEFENSIN1.2 expression was unaltered, indicating a repressor role for LBD20 in a branch of the JA-signaling pathway. Plants overexpressing LBD20 (LBD20-OX) had reduced Thi2.1 and VSP2 expression. There was a significant correlation between increased LBD20 expression in the LBD20-OX lines with both Thi2.1 and VSP2 repression, and reduced survival following F. oxysporum infection. Chlorosis resulting from application of F. oxysporum culture filtrate was also reduced in lbd20 leaves relative to the wild type. Taken together, LBD20 is a F. oxysporum susceptibility gene that appears to regulate components of JA signaling downstream of COI1 and MYC2 that are required for full elicitation of F. oxysporum- and JA-dependent responses. To our knowledge, this is the first demonstration of a role for a LBD gene family member in either biotic stress or JA signaling. PMID:22786889
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Ahmad, Aijaz; Khan, Amber; Manzoor, Nikhat
2013-01-23
Thymol (THY) and carvacrol (CARV), the principal chemical components of thyme oil have long been known for their wide use in medicine due to antimicrobial and disinfectant properties. This study, however, draws attention to a possible synergistic antifungal effect of these monoterpenes with azole antimycotic-fluconazole. Resistance to azoles in Candida albicans involves over-expression of efflux-pump genes MDR1, CDR1, CDR2 or mutations and over-expression of target gene ERG11. The inhibition of drug efflux pumps is considered a feasible strategy to overcome clinical antifungal resistance. To put forward this approach, we investigated the combination effects of these monoterpenes and FLC against 38 clinically obtained FLC-sensitive, and eleven FLC-resistant Candida isolates. Synergism was observed with combinations of THY-FLC and CARV-FLC evaluated by checkerboard microdilution method and nature of the interactions was calculated by FICI. In addition, antifungal activity was assessed using agar-diffusion and time-kill curves. The drug efflux activity was determined using two dyes, Rhodamine6G (R6G) and fluorescent Hoechst 33342. No significant differences were observed in dye uptakes between FLC-susceptible and resistant isolates, incubated in glucose free buffer. However, a significantly higher efflux was recorded in FLC-resistant isolates when glucose was added. Both monoterpenes inhibited efflux by 70-90%, showing their high potency to block drug transporter pumps. Significant differences, in the expression levels of CDR1 and MDR1, induced by monoterpenes revealed reversal of FLC-resistance. The selectively fungicidal characteristics and ability to restore FLC susceptibility in resistant isolates signify a promising candidature of THY and CARV as antifungal agents in combinational treatments for candidiasis. Copyright © 2012 Elsevier B.V. All rights reserved.
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Kim, Bong Hyun; Kim, Kyuseok; Nam, Hae Jeong
2017-01-31
Many previous studies of electroacupuncture used combined therapy of electroacupuncture and systemic manual acupuncture, so it was uncertain which treatment was effective. This study evaluated and compared the effects of systemic manual acupuncture, periauricular electroacupuncture and distal electroacupuncture for treating patients with tinnitus. A randomized, parallel, open-labeled exploratory trial was conducted. Subjects aged 20-75 years who had suffered from idiopathic tinnitus for > 2 weeks were recruited from May 2013 to April 2014. The subjects were divided into three groups by systemic manual acupuncture group (MA), periauricular electroacupuncture group (PE), and distal electroacupuncture group (DE). The groups were selected by random drawing. Nine acupoints (TE 17, TE21, SI19, GB2, GB8, ST36, ST37, TE3 and TE9), two periauricular acupoints (TE17 and TE21), and four distal acupoints (TE3, TE9, ST36, and ST37) were selected. The treatment sessions were performed twice weekly for a total of eight sessions over 4 weeks. Outcomes were the tinnitus handicap inventory (THI) score and the loud and uncomfortable visual analogue scales (VAS). Demographic and clinical characteristics of all participants were compared between the groups upon admission using one-way analysis of variance (ANOVA). One-way ANOVA was used to evaluate the THI, VAS loud , and VAS uncomfortable scores. The least significant difference test was used as a post-hoc test. Thirty-nine subjects were eligible and their data were analyzed. No difference in THI and VAS loudness scores was observed in between groups. The VAS uncomfortable scores decreased significantly in MA and DE compared with those in PE. Within the group, all three treatments showed some effect on THI, VAS loudness scores and VAS uncomfortable scores after treatment except DE in THI. There was no statistically significant difference between systemic manual acupuncture, periauricular electroacupuncture and distal electroacupuncture in tinnitus. However, all three treatments had some effect on tinnitus within the group before and after treatment. Systemic manual acupuncture and distal electroacupuncture have some effect on VAS uncomfortable . KCT0001991 by CRIS (Clinical Research Information Service), 2016-8-1, retrospectively registered.
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Technical note: estimating absorbed doses to the thyroid in CT.
Huda, Walter; Magill, Dennise; Spampinato, Maria V
2011-06-01
To describe a method for estimating absorbed doses to the thyroid in patients undergoing neck CT examinations. Thyroid doses in anthropomorphic phantoms were obtained for all 23 scanner dosimetry data sets in the ImPACT CT patient dosimetry calculator. Values of relative thyroid dose [R(thy)(L)], defined as the thyroid dose for a given scan length (L) divided by the corresponding thyroid dose for a whole body scan, were determined for neck CT scans. Ratios of the maximum thyroid dose to the corresponding CTDI(vol) and [D'(thy)], were obtained for two phantom diameters. The mass-equivalent water cylinder of any patient can be derived from the neck cross-sectional area and the corresponding average Hounsfield Unit, and compared to the 16.5-cm diameter water cylinder that models the ImPACT anthropomorphic phantom neck. Published values of relative doses in water cylinders of varying diameter were used to adjust thyroid doses in the anthropomorphic phantom to those of any sized patient. Relative thyroid doses R(thy)(L) increase to unity with increasing scan length and with very small difference between scanners. A 10-cm scan centered on the thyroid would result in a dose that is, nearly 90% of the thyroid dose from a whole body scan when performed using the constant radiographic techniques. At 120 kV, the average value of D'(thy) for the 16-cm diameter was 1.17 +/- 0.05 and was independent of CT vendor and year of CT scanner, and choice of x-ray tube voltage. The corresponding average value of D'(thy) in the 32-cm diameter phantom was 2.28 +/- 0.22 and showed marked variations depending on vendor, year of introduction into clinical practice as well as x-ray tube voltage. At 120 kV, a neck equivalent to a 10-cm diameter cylinder of water would have thyroid doses 36% higher than those in the ImPACT phantom, whereas a neck equivalent to a 25-cm cylinder diameter would have thyroid doses 35% lower. Patient thyroid doses can be estimated by taking into account the amount of radiation used to perform the CT examination (CTDI(vol)) and accounting for scan length and patient anatomy (i.e., neck diameter) at the thyroid location.
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Santana, Mário L; Bignardi, Annaiza Braga; Pereira, Rodrigo Junqueira; Menéndez-Buxadera, Alberto; El Faro, Lenira
2016-02-01
The present study had the following objectives: to compare random regression models (RRM) considering the time-dependent (days in milk, DIM) and/or temperature × humidity-dependent (THI) covariate for genetic evaluation; to identify the effect of genotype by environment interaction (G×E) due to heat stress on milk yield; and to quantify the loss of milk yield due to heat stress across lactation of cows under tropical conditions. A total of 937,771 test-day records from 3603 first lactations of Brazilian Holstein cows obtained between 2007 and 2013 were analyzed. An important reduction in milk yield due to heat stress was observed for THI values above 66 (-0.23 kg/day/THI). Three phases of milk yield loss were identified during lactation, the most damaging one at the end of lactation (-0.27 kg/day/THI). Using the most complex RRM, the additive genetic variance could be altered simultaneously as a function of both DIM and THI values. This model could be recommended for the genetic evaluation taking into account the effect of G×E. The response to selection in the comfort zone (THI ≤ 66) is expected to be higher than that obtained in the heat stress zone (THI > 66) of the animals. The genetic correlations between milk yield in the comfort and heat stress zones were less than unity at opposite extremes of the environmental gradient. Thus, the best animals for milk yield in the comfort zone are not necessarily the best in the zone of heat stress and, therefore, G×E due to heat stress should not be neglected in the genetic evaluation.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Lai, Rung-Yi; Huang, Siyu; Fenwick, Michael K.
2012-06-26
In Saccharomyces cerevisiae, thiamin pyrimidine is formed from histidine and pyridoxal phosphate (PLP). The origin of all of the pyrimidine atoms has been previously determined using labeling studies and suggests that the pyrimidine is formed using remarkable chemistry that is without chemical or biochemical precedent. Here we report the overexpression of the closely related Candida albicans pyrimidine synthase (THI5p) and the reconstitution and preliminary characterization of the enzymatic activity. A structure of the C. albicans THI5p shows PLP bound at the active site via an imine with Lys62 and His66 in close proximity to the PLP. Our data suggest thatmore » His66 of the THI5 protein is the histidine source for pyrimidine formation and that the pyrimidine synthase is a single-turnover enzyme.« less
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Zhou, Wenbo; Milder, Julie B; Freed, Curt R
2008-04-11
Abnormal aggregation of human alpha-synuclein in Lewy bodies and Lewy neurites is a pathological hallmark of Parkinson disease and dementia with Lewy bodies. Studies have shown that oxidation and nitration of alpha-synuclein lead to the formation of stable dimers and oligomers through dityrosine cross-linking. Previously we have reported that tyrosine-to-cysteine mutations, particularly at the tyrosine 39 residue (Y39C), significantly enhanced alpha-synuclein fibril formation and neurotoxicity. In the current study, we have generated transgenic mice expressing the Y39C mutant human alpha-synuclein gene controlled by the mouse Thy1 promoter. Mutant human alpha-synuclein was widely expressed in transgenic mouse brain, resulting in 150% overexpression relative to endogenous mouse alpha-synuclein. At age 9-12 months, transgenic mice began to display motor dysfunction in rotarod testing. Older animals aged 15-18 months showed progressive accumulation of human alpha-synuclein oligomers, associated with worse motor function and cognitive impairment in the Morris water maze. By age 21-24 months, alpha-synuclein aggregates were further increased, accompanied by severe behavioral deficits. At this age, transgenic mice developed neuropathology, such as Lewy body-like alpha-synuclein and ubiquitin-positive inclusions, phosphorylation at Ser(129) of human alpha-synuclein, and increased apoptotic cell death. In summary, Y39C human alpha-synuclein transgenic mice show age-dependent, progressive neuronal degeneration with motor and cognitive deficits similar to diffuse Lewy body disease. The time course of alpha-synuclein oligomer accumulation coincided with behavioral and pathological changes, indicating that these oligomers may initiate protein aggregation, disrupt cellular function, and eventually lead to neuronal death.
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Ghaderi, Shima; Soheili, Zahra-Soheila; Ahmadieh, Hamid; Davari, Maliheh; Jahromi, Fatemeh Sanie; Samie, Shahram; Rezaie-Kanavi, Mozhgan; Pakravesh, Jalil; Deezagi, Abdolkhalegh
2011-09-01
To evaluate the effect of human amniotic fluid (HAF) on retinal pigmented epithelial cells growth and trans-differentiation into retinal neurons, retinal pigmented epithelium (RPE) cells were isolated from neonatal human cadaver eye globes and cultured in Dulbecco's modified Eagle's medium-F12 supplemented with 10% fetal bovine serum (FBS). Confluent monolayer cultures were trypsinized and passaged using FBS-containing or HAF-containing media. Amniotic fluid samples were received from pregnant women in the first trimester of gestation. Cell proliferation and death enzyme-linked immunosorbent assays were performed to assess the effect of HAF on RPE cell growth. Trans-differentiation into rod photoreceptors and retinal ganglion cells was also studied using immunocytochemistry and real-time polymerase chain reaction techniques. Primary cultures of RPE cells were successfully established under FBS-containing or HAF-containing media leading to rapid cell growth and proliferation. When RPE cells were moved to in vitro culture system, they began to lose their differentiation markers such as pigmentation and RPE65 marker and trans-differentiated neural-like cells followed by spheroid colonies pertaining to stem/progenitor cells were morphologically detected. Immunocytochemistry (ICC) analysis of HAF-treated cultures showed a considerable expression of Rhodopsin gene (30% Rhodopsin-positive cells) indicating trans-differentiation of RPE cells to rod photoreceptors. Real-time polymerase chain reaction revealed an HAF-dose-dependant expression of Thy-1 gene (RGC marker) and significant promoting effect of HAF on RGCs generation. The data presented here suggest that HAF possesses invaluable stimulatory effect on RPE cells growth and trans-differentiation into retinal neurons. It can be regarded as a newly introduced enriched supplement in serum-free kinds of media used in neuro-retinal regeneration studies.
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Ray, Radharaman; Zhang, Peng; Ray, Prabhati
2011-08-01
The passion in the scientific endeavors of Marshall Warren Nirenberg had been his quest for knowledge regarding the storage, retrieval, and processing of information in the cell. After deciphering the genetic code for which he shared the Nobel Prize in Physiology and Medicine in 1968, Nirenberg devoted his attention to unraveling the mysteries in the most complex cellular organization in the body, i.e., the nervous system, especially those governing neuronal development, plasticity, and synaptogenesis. During the tenure of the primary author (RR) as a postdoctoral Staff Fellow in the Nirenberg laboratory in the late seventies to early eighties, he had the opportunity of working on projects related to what Nirenberg used to broadly define as the "synaptic code." The major aspects of these projects dealt with the functional macromolecules relevant to neuronal growth, organization, lineage, selectivity, stabilization, synaptogenesis, and functions such as neuroexocytosis. This author's emphasis was particularly on voltage-gated calcium channels that regulate stimulus-induced neurotransmitter release. One central as well as crucial theme in these studies was the fact that the neurons had to be mature and differentiated in order to study these entities (Science 222: 794-799, 1983; Cold Spring Harb Symp Quant Biol 48: 707-715, 1983). In this communication, we illustrate how did this basic knowledge, i.e., cell maturation-dependent properties being essential for neuronal functions, led to a successful experimental design and demonstration of the validity of the targeted neurologic therapeutic delivery approach based on recombinant botulinum toxin serotype A (BoNT/A) heavy chain (rHC) serving as a neuron-specific targeting molecule (BMC Pharmacol 9: 12, 2009).
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Ronzitti, Emiliano; Conti, Rossella; Zampini, Valeria; Tanese, Dimitrii; Klapoetke, Nathan; Boyden, Edward S.; Papagiakoumou, Eirini
2017-01-01
Optogenetic neuronal network manipulation promises to unravel a long-standing mystery in neuroscience: how does microcircuit activity relate causally to behavioral and pathological states? The challenge to evoke spikes with high spatial and temporal complexity necessitates further joint development of light-delivery approaches and custom opsins. Two-photon (2P) light-targeting strategies demonstrated in-depth generation of action potentials in photosensitive neurons both in vitro and in vivo, but thus far lack the temporal precision necessary to induce precisely timed spiking events. Here, we show that efficient current integration enabled by 2P holographic amplified laser illumination of Chronos, a highly light-sensitive and fast opsin, can evoke spikes with submillisecond precision and repeated firing up to 100 Hz in brain slices from Swiss male mice. These results pave the way for optogenetic manipulation with the spatial and temporal sophistication necessary to mimic natural microcircuit activity. SIGNIFICANCE STATEMENT To reveal causal links between neuronal activity and behavior, it is necessary to develop experimental strategies to induce spatially and temporally sophisticated perturbation of network microcircuits. Two-photon computer generated holography (2P-CGH) recently demonstrated 3D optogenetic control of selected pools of neurons with single-cell accuracy in depth in the brain. Here, we show that exciting the fast opsin Chronos with amplified laser 2P-CGH enables cellular-resolution targeting with unprecedented temporal control, driving spiking up to 100 Hz with submillisecond onset precision using low laser power densities. This system achieves a unique combination of spatial flexibility and temporal precision needed to pattern optogenetically inputs that mimic natural neuronal network activity patterns. PMID:28972125
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The Sense of Smell Impacts Metabolic Health and Obesity.
Riera, Celine E; Tsaousidou, Eva; Halloran, Jonathan; Follett, Patricia; Hahn, Oliver; Pereira, Mafalda M A; Ruud, Linda Engström; Alber, Jens; Tharp, Kevin; Anderson, Courtney M; Brönneke, Hella; Hampel, Brigitte; Filho, Carlos Daniel de Magalhaes; Stahl, Andreas; Brüning, Jens C; Dillin, Andrew
2017-07-05
Olfactory inputs help coordinate food appreciation and selection, but their role in systemic physiology and energy balance is poorly understood. Here we demonstrate that mice upon conditional ablation of mature olfactory sensory neurons (OSNs) are resistant to diet-induced obesity accompanied by increased thermogenesis in brown and inguinal fat depots. Acute loss of smell perception after obesity onset not only abrogated further weight gain but also improved fat mass and insulin resistance. Reduced olfactory input stimulates sympathetic nerve activity, resulting in activation of β-adrenergic receptors on white and brown adipocytes to promote lipolysis. Conversely, conditional ablation of the IGF1 receptor in OSNs enhances olfactory performance in mice and leads to increased adiposity and insulin resistance. These findings unravel a new bidirectional function for the olfactory system in controlling energy homeostasis in response to sensory and hormonal signals. Copyright © 2017 Elsevier Inc. All rights reserved.
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Lee, Hongje; Lee, Ho Won; La Lee, You; Jeon, Yong Hyun; Jeong, Shin Young; Lee, Sang-Woo; Lee, Jaetae; Ahn, Byeong-Cheol
2018-06-01
The aim of this study is to optimize the dendritic cell (DC)-mediated T-cell activation using reporter gene imaging and flow cytometric analysis in living mice. A murine dendritic cell line (DC2.4) co-expressing effluc and Thy1.1 genes were established by transfection with retroviral vectors. Thy1.1 positive cells were sorted by magnetic bead separation system (DC2.4/effluc). Cell proliferation assay and phenotype analysis to determine the effects of gene transduction on the function of dendritic cells between parental DC2.4 and DC2.4/effluc were performed. To optimize the DC-mediated immune response by cell number or frequency, different cell numbers (5 × 10 5 , 1 × 10 6 , and 2 × 10 6 DC2.4/effluc) or different frequencies of DC2.4/effluc (first, second, and third injections) were injected in the right footpad of mice. The migration of the DC2.4/effluc into the draining popliteal lymph node of mice was monitored by bioluminescence imaging (BLI). Flow cytometric analysis was performed with splenocytes to determine the cytotoxic T-cell population after injection of DC2.4/effluc. Parental DC2.4 and DC2.4/effluc exhibit no significant differences in their proliferation and phenotype. BLI signals were observed in the draining popliteal lymph node at day 1 after injection of DC2.4/effluc in 1 × 10 6 and 2 × 10 6 cells-injected groups. The highest BLI signal intensity was detected in 2 × 10 6 cells-injected mice. On day 11, the BLI signal was detected in only 2 × 10 6 cell-injected group but not in other groups. Optimized cell numbers (2 × 10 6 ) were injected in three animal groups with a different frequency (first, second, and third injection groups). The BLI signal was detected at day 1 and maintained until day 7 in the first injection group, but there is low signal intensity in the second and the third injection groups. Although the expression levels of Thy1.1 gene in the first injection group were very high, there reveals no expression of Thy1.1 gene in the second and the third injection groups. The number of tumor-specific CD8 + T-cells in the spleen significantly increased, as the number of DC injections increases. Successful optimization of DC-mediated cytotoxic T-cell activation in living mice using reporter gene imaging and flow cytometric analysis was achieved. The optimization of DC-mediated cytotoxic T-cell activation could be applied for the future DC-based immunotherapy.
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ODE Constrained Mixture Modelling: A Method for Unraveling Subpopulation Structures and Dynamics
Hasenauer, Jan; Hasenauer, Christine; Hucho, Tim; Theis, Fabian J.
2014-01-01
Functional cell-to-cell variability is ubiquitous in multicellular organisms as well as bacterial populations. Even genetically identical cells of the same cell type can respond differently to identical stimuli. Methods have been developed to analyse heterogeneous populations, e.g., mixture models and stochastic population models. The available methods are, however, either incapable of simultaneously analysing different experimental conditions or are computationally demanding and difficult to apply. Furthermore, they do not account for biological information available in the literature. To overcome disadvantages of existing methods, we combine mixture models and ordinary differential equation (ODE) models. The ODE models provide a mechanistic description of the underlying processes while mixture models provide an easy way to capture variability. In a simulation study, we show that the class of ODE constrained mixture models can unravel the subpopulation structure and determine the sources of cell-to-cell variability. In addition, the method provides reliable estimates for kinetic rates and subpopulation characteristics. We use ODE constrained mixture modelling to study NGF-induced Erk1/2 phosphorylation in primary sensory neurones, a process relevant in inflammatory and neuropathic pain. We propose a mechanistic pathway model for this process and reconstructed static and dynamical subpopulation characteristics across experimental conditions. We validate the model predictions experimentally, which verifies the capabilities of ODE constrained mixture models. These results illustrate that ODE constrained mixture models can reveal novel mechanistic insights and possess a high sensitivity. PMID:24992156
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Reentrant Phase Coherence in Superconducting Nanowire Composites.
Ansermet, Diane; Petrović, Alexander P; He, Shikun; Chernyshov, Dmitri; Hoesch, Moritz; Salloum, Diala; Gougeon, Patrick; Potel, Michel; Boeri, Lilia; Andersen, Ole Krogh; Panagopoulos, Christos
2016-01-26
The short coherence lengths characteristic of low-dimensional superconductors are associated with usefully high critical fields or temperatures. Unfortunately, such materials are often sensitive to disorder and suffer from phase fluctuations in the superconducting order parameter which diverge with temperature T, magnetic field H, or current I. We propose an approach to overcome synthesis and fluctuation problems: building superconductors from inhomogeneous composites of nanofilaments. Macroscopic crystals of quasi-one-dimensional Na2-δMo6Se6 featuring Na vacancy disorder (δ ≈ 0.2) are shown to behave as percolative networks of superconducting nanowires. Long-range order is established via transverse coupling between individual one-dimensional filaments, yet phase coherence remains unstable to fluctuations and localization in the zero (T,H,I) limit. However, a region of reentrant phase coherence develops upon raising (T,H,I). We attribute this phenomenon to an enhancement of the transverse coupling due to electron delocalization. Our observations of reentrant phase coherence coincide with a peak in the Josephson energy EJ at nonzero (T,H,I), which we estimate using a simple analytical model for a disordered anisotropic superconductor. Na2-δMo6Se6 is therefore a blueprint for a future generation of nanofilamentary superconductors with inbuilt resilience to phase fluctuations at elevated (T,H,I).
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Physics of tissue harmonic imaging by ultrasound
NASA Astrophysics Data System (ADS)
Jing, Yuan
Tissue Harmonic Imaging (THI) is an imaging modality that is currently deployed on diagnostic ultrasound scanners. In THI the amplitude of the ultrasonic pulse that is used to probe the tissue is large enough that the pulse undergoes nonlinear distortion as it propagates into the tissue. One result of the distortion is that as the pulse propagates energy is shifted from the fundamental frequency of the source pulse into its higher harmonics. These harmonics will scatter off objects in the tissue and images formed from the scattered higher harmonics are considered to have superior quality to the images formed from the fundamental frequency. Processes that have been suggested as possibly responsible for the improved imaging in THI include: (1) reduced sensitivity to reverberation, (2) reduced sensitivity to aberration, and (3) reduction in side lobes. By using a combination of controlled experiments and numerical simulations, these three reasons have been investigated. A single element transducer and a clinical ultrasound scanner with a phased array transducer were used to image a commercial tissue-mimicking phantom with calibrated targets. The higher image quality achieved with THI was quantified in terms of spatial resolution and "clutter" signals. A three-dimensional model of the forward propagation of nonlinear sound beams in media with arbitrary spatial properties (a generalized KZK equation) was developed. A time-domain code for solving the KZK equation was validated with measurements of the acoustic field generated by the single element transducer and the phased array transducer. The code was used to investigate the impact of aberration using tissue-like media with three-dimensional variations in all acoustic properties. The three-dimensional maps of tissue properties were derived from the datasets available through the Visible Female project. The experiments and simulations demonstrated that second harmonic imaging (1) suffers less clutter associated with reverberation; (2) is not immune to aberration effects and (3) suffers less clutter due to reduced side-lobe levels. The results indicate that side lobe suppression is the most significant reason for the improvement of second harmonic imaging.
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Lack of Diaph3 relaxes the spindle checkpoint causing the loss of neural progenitors
Damiani, Devid; Goffinet, André M.; Alberts, Arthur; Tissir, Fadel
2016-01-01
The diaphanous homologue Diaph3 (aka mDia2) is a major regulator of actin cytoskeleton. Loss of Diaph3 has been constantly associated with cytokinesis failure ascribed to impaired accumulation of actin in the cleavage furrow. Here we report that Diaph3 is required before cell fission, to ensure the accurate segregation of chromosomes. Inactivation of the Diaph3 gene causes a massive loss of cortical progenitor cells, with subsequent depletion of intermediate progenitors and neurons, and results in microcephaly. In embryonic brain extracts, Diaph3 co-immunoprecipitates with BubR1, a key regulator of the spindle assembly checkpoint (SAC). Diaph3-deficient cortical progenitors have decreased levels of BubR1 and fail to properly activate the SAC. Hence, they bypass mitotic arrest and embark on anaphase in spite of incorrect chromosome segregation, generating aneuploidy. Our data identify Diaph3 as a major guard of cortical progenitors, unravel novel functions of Diaphanous formins and add insights into the pathobiology of microcephaly. PMID:27848932
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The DPP4 Inhibitor Linagliptin Protects from Experimental Diabetic Retinopathy.
Dietrich, Nadine; Kolibabka, Matthias; Busch, Stephanie; Bugert, Petra; Kaiser, Ulrike; Lin, Jihong; Fleming, Thomas; Morcos, Michael; Klein, Thomas; Schlotterer, Andrea; Hammes, Hans-Peter
2016-01-01
Dipeptidyl peptidase 4 (DPP4) inhibitors improve glycemic control in type 2 diabetes, however, their influence on the retinal neurovascular unit remains unclear. Vasculo- and neuroprotective effects were assessed in experimental diabetic retinopathy and high glucose-cultivated C. elegans, respectively. In STZ-diabetic Wistar rats (diabetes duration of 24 weeks), DPP4 activity (fluorometric assay), GLP-1 (ELISA), methylglyoxal (LC-MS/MS), acellular capillaries and pericytes (quantitative retinal morphometry), SDF-1a and heme oxygenase-1 (ELISA), HMGB-1, Iba1 and Thy1.1 (immunohistochemistry), nuclei in the ganglion cell layer, GFAP (western blot), and IL-1beta, Icam1, Cxcr4, catalase and beta-actin (quantitative RT-PCR) were determined. In C. elegans, neuronal function was determined using worm tracking software. Linagliptin decreased DPP4 activity by 77% and resulted in an 11.5-fold increase in active GLP-1. Blood glucose and HbA1c were reduced by 13% and 14% and retinal methylglyoxal by 66%. The increase in acellular capillaries was diminished by 70% and linagliptin prevented the loss of pericytes and retinal ganglion cells. The rise in Iba-1 positive microglia was reduced by 73% with linagliptin. In addition, the increase in retinal Il1b expression was decreased by 65%. As a functional correlate, impairment of motility (body bending frequency) was significantly prevented in C. elegans. Our data suggest that linagliptin has a protective effect on the microvasculature of the diabetic retina, most likely due to a combination of neuroprotective and antioxidative effects of linagliptin on the neurovascular unit.
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Novel astrocyte targets: new avenues for the therapeutic treatment of epilepsy.
Crunelli, Vincenzo; Carmignoto, Giorgio; Steinhäuser, Christian
2015-02-01
During the last 20 years, it has been well established that a finely tuned, continuous crosstalk between neurons and astrocytes not only critically modulates physiological brain functions but also underlies many neurological diseases. In particular, this novel way of interpreting brain activity is markedly influencing our current knowledge of epilepsy, prompting a re-evaluation of old findings and guiding novel experimentation. Here, we review recent studies that have unraveled novel and unique contributions of astrocytes to the generation and spread of convulsive and nonconvulsive seizures and epileptiform activity. The emerging scenario advocates an overall framework in which a dynamic and reciprocal interplay among astrocytic and neuronal ensembles is fundamental for a fuller understanding of epilepsy. In turn, this offers novel astrocytic targets for the development of those really novel chemical entities for the control of convulsive and nonconvulsive seizures that have been acknowledged as a key priority in the management of epilepsy. © The Author(s) 2014.
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Redox Regulation in Amyotrophic Lateral Sclerosis
Parakh, Sonam; Spencer, Damian M.; Halloran, Mark A.; Soo, Kai Y.; Atkin, Julie D.
2013-01-01
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that results from the death of upper and lower motor neurons. Due to a lack of effective treatment, it is imperative to understand the underlying mechanisms and processes involved in disease progression. Regulations in cellular reduction/oxidation (redox) processes are being increasingly implicated in disease. Here we discuss the possible involvement of redox dysregulation in the pathophysiology of ALS, either as a cause of cellular abnormalities or a consequence. We focus on its possible role in oxidative stress, protein misfolding, glutamate excitotoxicity, lipid peroxidation and cholesterol esterification, mitochondrial dysfunction, impaired axonal transport and neurofilament aggregation, autophagic stress, and endoplasmic reticulum (ER) stress. We also speculate that an ER chaperone protein disulphide isomerase (PDI) could play a key role in this dysregulation. PDI is essential for normal protein folding by oxidation and reduction of disulphide bonds, and hence any disruption to this process may have consequences for motor neurons. Addressing the mechanism underlying redox regulation and dysregulation may therefore help to unravel the molecular mechanism involved in ALS. PMID:23533690
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Odor Coding by a Mammalian Receptor Repertoire
Saito, Harumi; Chi, Qiuyi; Zhuang, Hanyi; Matsunami, Hiro; Mainland, Joel D.
2009-01-01
Deciphering olfactory encoding requires a thorough description of the ligands that activate each odorant receptor (OR). In mammalian systems, however, ligands are known for fewer than 50 of over 1400 human and mouse ORs, greatly limiting our understanding of olfactory coding. We performed high-throughput screening of 93 odorants against 464 ORs expressed in heterologous cells and identified agonists for 52 mouse and 10 human ORs. We used the resulting interaction profiles to develop a predictive model relating physicochemical odorant properties, OR sequences, and their interactions. Our results provide a basis for translating odorants into receptor neuron responses and unraveling mammalian odor coding. PMID:19261596
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Artificial Intelligence Technical Paper Abstracts 1991
1992-07-08
f se118urca,. gathering and inaitaing the data nee ded. and completin ene reuewmng the collection f informtio sn continnt rey dsng thi bude estimate...89) Pr’fS o b’ ANS I Sid 139-1 19 0 Table of Contents Introduction
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Red blood cell transfusions and tissue oxygenation in anemic hematology outpatients.
Yuruk, Koray; Bartels, Sebastiaan A; Milstein, Dan M J; Bezemer, Rick; Biemond, Bart J; Ince, Can
2012-03-01
There is little clinical evidence that red blood cell (RBC) transfusions improve oxygen availability at the microcirculatory level. We tested the hypotheses that anemia in chronically anemic patients with relatively healthy microcirculation would be associated with low tissue hemoglobin (Hb) and tissue oxygenation levels and that these conditions would be improved after RBC transfusions. Near-infrared spectroscopy (NIRS) was used to determine tissue oxygen saturation (StO(2)) and tissue Hb index (THI; an index of the amount of Hb in the NIRS measurement volume) in the thenar eminence and sublingual tissue before and 30 minutes after RBC transfusions in 20 chronically anemic hematology outpatients. Data are presented as median (25%-75%). The patients received three (two to three) bags of RBCs in saline-adenine-glucose-mannitol with an age of 21 (7-21) days, which was infused intravenously at the rate of 0.7 bag/hr. RBC transfusions significantly increased hematocrit level from 26% (24%-28%) to 32% (30%-34%; p < 0.0001), Hb level from 8.2 (7.6-8.9) g/dL to 11.0 (9.9-11.8) g/dL (p < 0.0001), whole blood viscosity from 3.4 (3.1-3.5) mPa/sec to 4.2 (4.0-4.5) mPa/sec (p < 0.0001), thenar StO(2) from 81% (80%-84%) to 86% (81%-89%; p = 0.002), thenar THI from 11.2 (9.3-13.3) AU to 13.7 (9.7-15.3) AU (p = 0.024), sublingual StO(2) from 86% (81%-89%) to 91% (86%-92%; p < 0.0001), and sublingual THI from 15.2 (13.0-17.4) AU to 17.2 (13.5-19.7) AU (p = 0.040). Although anemia in chronically anemic hematology outpatients was not associated with low StO(2) and THI levels, RBC transfusions were successful in improving these variables. © 2011 American Association of Blood Banks.
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Auditory discrimination therapy (ADT) for tinnitus management.
Herraiz, C; Diges, I; Cobo, P
2007-01-01
Auditory discrimination training (ADT) designs a procedure to increase cortical areas responding to trained frequencies (damaged cochlear areas with cortical misrepresentation) and to shrink the neighboring over-represented ones (tinnitus pitch). In a prospective descriptive study of 27 patients with high frequency tinnitus, the severity of the tinnitus was measured using a visual analog scale (VAS) and the tinnitus handicap inventory (THI). Patients performed a 10-min auditory discrimination task twice a day during one month. Discontinuous 4 kHz pure tones were mixed randomly with short broadband noise sounds through an MP3 system. After the treatment mean VAS scores were reduced from 5.2 to 4.5 (p=0.000) and the THI decreased from 26.2% to 21.3% (p=0.000). Forty percent of the patients had improvement in tinnitus perception (RESP). Comparing the ADT group with a control group showed statistically significant improvement of their tinnitus as assessed by RESP, VAS, and THI.
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Carneiro, Cláudia Martins; Martins-Filho, Olindo Assis; Reis, Alexandre Barbosa; Veloso, Vanja Maria; Araújo, Flávio Marcos Gomes; Bahia, Maria Terezinha; de Lana, Marta; Machado-Coelho, George Luiz Lins; Gazzinelli, Giovanni; Correa-Oliveira, Rodrigo; Tafuri, Washington Luiz
2007-02-01
A detailed follow-up investigation of the major parasitological, serological and phenotypic features in dogs experimentally infected with metacyclic (MT) and blood (BT) trypomastigotes of Trypanosoma cruzi strain Berenice-78, typifying vectorial and transfusional transmission of human Chagas disease, has been conducted. Although there were no changes with respect to the window of patent-parasitaemia, significant differences between MT- and BT-infected dogs in both the prepatent period (days 23 and 19, respectively) and the day of maximum parasitaemia (days 26 and 22, respectively) were recorded. A progressive enhancement in the level of T. cruzi-specific antibodies accompanied infection by both MT and BT forms, although higher IgG titres developed on days 14 and 21 following infection with MT forms. Higher Thy-1(+)/CD21(+) and lower CD4(+)/CD8(+) cell ratios, occasioned by increased levels of Thy-1(+) and CD8(+) T-cells and reduced frequencies of CD4(+) T-cells and CD21(+) B-lymphocytes, were observed in both MT- and BT-infected animals. The reduced frequency of CD14(+) leukocytes was revealed as the most relevant phenotypic feature intrinsic to T. cruzi infection independent of inoculum source. BT-specific phenotypic features included an early reduction in the percentage of circulating CD21(+) and CD14(+) leukocytes, together with a higher Thy-1(+)/CD21(+) cell ratio on day 42. On the other hand, higher levels of CD8(+) T-cells, together with a lower CD4(+)/CD8(+) cell ratio on day 28, were characteristic of MT infection. These findings emphasise the importance of inoculum source and suggest that vectorial or transfusional routes of T. cruzi infection may trigger distinct parasite-host interactions during acute Chagas disease.
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Trailer microclimate and calf welfare during fall-run transportation of beef calves in Alberta.
Goldhawk, C; Janzen, E; González, L A; Crowe, T; Kastelic, J; Pajor, E; Schwartzkopf-Genswein, K S
2014-11-01
Twenty-four commercial loads of beef calves (BW 300 ± 52 kg, mean ± SD) were evaluated for associations among transportation factors, in-transit microclimate, and calf welfare. Transport factors evaluated included vehicle speed, space allowance, compartment within trailer, and transit duration. Calves were transported for 7 h 44 min ± 4 h 15 min, with space allowances ranging from 0.56 to 1.17 m(2)/animal. Compartment within trailer, space allowance, and vehicle speed did not affect the difference between compartment ceiling-level and ambient temperatures during a 30-min period of steady-state microclimate. During the steady-state period, a 1°C increase in ambient temperature above the mean of 5.6°C was associated with a 0.62°C decrease in the difference between ceiling-level and ambient temperature (P < 0.01). Ceiling-level temperature and humidity during the first 400 min of transport could be predicted by ambient conditions and vehicle speed (pseudo-r(2) of 0.91 and 0.82 for temperature and humidity ratio; P < 0.01). Events when animal-level temperature-humidity index (THI) was classified as above the "danger" level lasted for 10.2 ± 4.1 consecutive minutes. Ambient and ceiling-level THI values were not classified as above "danger" for 90.0 and 84.9% of animal-level events. Ambient and ceiling-level THI were 5.0 ± 2.1 and 4.7 ± 2.0° Flower than animal-level THI during periods of disagreement, respectively. The majority of calves arrived in good condition and biochemical indicators of calf welfare were within reference ranges for healthy cattle. Within the study population, high pre-transport cortisol and hematocrit were associated with elevated post-transport values (P < 0.01). A 1% increase in shrink during the weaning to loading interval (24 or 48 h) decreased transportation shrink by 0.26 ± 0.04% when average animal-level temperature was greater than 5°C and decreased transportation shrink by 0.11 ± 0.04% when average animal-level temperature was less than 5°C (P < 0.01). We inferred that the study results support future investigation of the extension of in-transit microclimate as a risk factor for post-transport treatment for disease. The study also provided correction factors for estimating in-transit microclimate that could assist in evaluation of transportation management and decisions affecting profitability and calf welfare.
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Mahboubi, Hossein; Haidar, Yarah M; Kiumehr, Saman; Ziai, Kasra; Djalilian, Hamid R
2017-10-01
To determine the effectiveness of a customized sound therapy and compare its effectiveness to that of masking with broadband noise. Subjects were randomized to receive either customized sound therapy or broadband noise for 2 hours per day for 3 months and then switched to the other treatment after a washout period. The outcome variables were tinnitus loudness (scored 0-10), Tinnitus Handicap Inventory (THI), Beck Anxiety Inventory (BAI), minimum masking levels (MML), and residual inhibition (RI). Eighteen subjects completed the study. Mean age was 53 ± 11 years, and mean tinnitus duration was 118 ± 99 months. With customized sound therapy, mean loudness decreased from 6.4 ± 2.0 to 4.9 ± 1.9 ( P = .001), mean THI decreased from 42.8 ± 21.6 to 31.5 ± 20.3 ( P < .001), mean BAI decreased from 10.6 ± 10.9 to 8.3 ± 9.9 ( P = .01), and MML decreased from 22.3 ± 11.6 dB SL to 17.2 ± 10.6 dB SL ( P = .005). After 3 months of broadband noise therapy, only BAI and, to a lesser degree, MML decreased ( P = .003 and .04, respectively). Customized sound therapy can decrease the loudness and THI scores of tinnitus patients, and the results may be superior to broadband noise.
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Prevalence of tinnitus in workers exposed to noise and organophosphates
Delecrode, Camila Ribas; de Freitas, Thais Domingues; Frizzo, Ana Claúdia Figueiredo; Cardoso, Ana Claúdia Vieira
2012-01-01
Summary Introduction: Research on the workplace has emphasized the effects of noise exposure on workers' hearing, but has not considered the effects of agrochemicals. Aim: To evaluate and correlate the hearing level and tinnitus of workers exposed simultaneously to noise and organophosphates in their workplace and to measure tinnitus distress on their quality of life. Method: A retrospective clinical study. We evaluated 82 organophosphate sprinklers from the São Paulo State Regional Superintendence who were active in the fight against dengue and who were exposed to noise and organophosphates. We performed pure tone audiometry and applied the translated THI (Tinnitus Handicap Inventory) questionnaire. Results: Of the sample, 28.05% reported current tinnitus or had presented tinnitus, and the workers with tinnitus had an increased incidence of abnormal audiometry. The average hearing threshold for the 4–8-kHz frequency range of the workers with current tinnitus was higher than that of the others, and was most affected at the 4-kHz frequency. The THI score ranged 0–84, with an average score of 13.1. Twelve (52.17%) workers had THI scores consistent with discrete handicap. Conclusion: There is an increased incidence of abnormal pure tone audiometry in workers with tinnitus, and its impact on the workers' quality of life was discrete. The correlation between average hearing threshold and tinnitus distress was weak. PMID:25991953
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Auditory discrimination therapy (ADT) for tinnitus managment: preliminary results.
Herraiz, C; Diges, I; Cobo, P; Plaza, G; Aparicio, J M
2006-12-01
This clinical assay has demonstrated the efficacy of auditory discrimination therapy (ADT) in tinnitus management compared with a waiting-list group. In all, 43% of the ADT patients improved their tinnitus, and its intensity together with its handicap were statistically decreased (EMB rating: B-2). To describe the effect of sound discrimination training on tinnitus. ADT designs a procedure to increase the cortical representation of trained frequencies (damaged cochlear areas with a secondary reduction of cortical stimulation) and to shrink the neighbouring over-represented ones (corresponding to tinnitus pitch). This prospective descriptive study included 14 patients with high frequency matched tinnitus. Tinnitus severity was measured according to a visual analogue scale (VAS) and the Tinnitus Handicap Inventory (THI). Patients performed a 10-min auditory discrimination task twice a day for 1 month. Discontinuous 8 kHz pure tones were randomly mixed with 500 ms 'white noise' sounds through a MP3 system. ADT group results were compared with a waiting-list group (n=21). In all, 43% of our patients had improvement in their tinnitus. A significant improvement in VAS (p=0.004) and THI mean scores was achieved (p=0.038). Statistical differences between ADT and the waiting-list group have been proved, considering patients' self-evaluations (p=0.043) and VAS scores (p=0.004). A non-significant reduction of THI was achieved (p=0.113).
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Park, Sean S; Grills, Inga Siiner; Bojrab, Dennis; Pieper, Daniel; Kartush, Jack; Maitz, Ann; Martin, Arturo; Perez, Evelyn; Hahn, Yoav; Ye, Hong; Martinez, Alvaro; Chen, Peter
2011-06-01
To prospectively assess the quality of life (QOL) and hearing acuity in vestibular schwannoma (VS) patients after gamma knife surgery (GKS). Fifty-nine VS patients. GKS. Prospective follow-up algorithm included 36-item Short Form Health Survey (SF-36), Hearing Handicap Inventory (HHI), Dizziness Handicap Inventory (DHI), Tinnitus Handicap Inventory (THI), pure-tone average, and speech discrimination hearing scores (Gardner-Robertson and American Academy of Otolaryngology), performed before and after GKS at 1-, 3-, 6-, 12-, and 18-month posttreatment intervals. From December 2006 to November 2008, 59 VS patients were treated with a median follow-up of 15 months. At baseline, mean scores for SF-36, HHI, DHI, and THI were 73, 37, 17, and 23, respectively. Median baseline Gardner-Robertson and American Academy of Otolaryngology hearing acuity scores were 2 and B, respectively. No significant decline in SF-36 health survey was noted after GKS. Mean SF-36 score at baseline was 73, compared with a range of 70 to 77 at predetermined posttreatment intervals. Similarly, no significant changes in DHI, HHI, and THI were noted. Approximately 47% of patients with baseline serviceable hearing maintained serviceable hearing at 12 months. Significant acute and chronic worsening in hearing acuity were noted at 1 and 18 months, respectively. No correlative decline in QOL was noted as assessed by SF-36 or HHI. No significant decline in global QOL occurred after GKS with relatively short follow-up and approximately 50% survey completion. When discussing therapy options with VS patients, anticipated treatment-related QOL outcomes should be considered.
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KiSS-1 and reproduction: focus on its role in the metabolic regulation of fertility.
Tena-Sempere, Manuel
2006-01-01
Unraveling of the master role of kisspeptins, the products of the KiSS-1 gene, and their receptor, GPR54, in the control of reproduction has been a major breakthrough in contemporary neuroendocrinology. Indeed, since the disclosure of their reproductive dimension in late 2003, an ever-growing number of genetic, molecular, physiologic and pharmacological studies have defined the crucial role of KiSS-1 neurons as central processors for the dynamic regulation of the gonadotropic axis and its full activation at puberty. Yet, the potential role of the hypothalamic KiSS-1 system as an intermediary factor for the well-known interplay between energy status and reproduction initially received little attention. Recent data, however, strongly suggest a prominent role of KiSS-1 in the metabolic control of fertility, as expression of KiSS-1 gene at the hypothalamus is down-regulated in conditions of negative energy balance and kisspeptin administration is capable of overcoming the hypogonadotropic state observed in undernutrition and disturbed metabolic conditions. Leptin, the adipocyte hormone signaling the size of body energy stores, is likely to play a pivotal role in the metabolic control of the KiSS-1 system, since kisspeptin neurons express leptin receptors and leptin is able to normalize defective KiSS-1 gene expression in models of impaired gonadotropin secretion linked to hypoleptinemia, such as the ob/ob mouse and streptozotocin-induced diabetic rat. In sum, these data provide strong evidence for a central role of kisspeptins and GPR54 as molecular conduits for the metabolic regulation of reproductive function - a phenomenon with potential physiopathologic and therapeutic implications.
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Using historical data to measure transportation infrastructure constraints on land use.
DOT National Transportation Integrated Search
1998-06-01
This study had three goals: (1) To develop a method for reversing the planning process, such that we begin with transportation system usage and conclude with transportation system usage and conclude with an indication of land use; (2) To validate thi...
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Sun, Xia; Cao, Yaoyao; Gong, Zhili; Wang, Xiangyou; Zhang, Yan; Gao, Jinmei
2012-01-01
In this work, a novel amperometric immunosensor based on multi-walled carbon nanotubes-thionine-chitosan (MWCNTs-THI-CHIT) nanocomposite film as electrode modified material was developed for the detection of chlorpyrifos residues. The nanocomposite film was dropped onto a glassy carbon electrode (GCE), and then the anti-chlorpyrifos monoclonal antibody was covalently immobilized onto the surface of MWCNTs-THI-CHIT/GCE using the crosslinking agent glutaraldehyde (GA). The modification procedure was characterized by using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under the optimized conditions, a linear relationship between the relative change in peak current of different pulse voltammetry (DPV) and the logarithm of chlorpyrifos solution concentration was obtained in the range from 0.1 to 1.0 × 105 ng/mL with a detection limit of 0.046 ng/mL. The proposed chlorpyrifos immunosensor exhibited high reproducibility, stability, and good selectivity and regeneration, making it a potential alternative tool for ultrasensitive detection of chlorpyrifos residues in vegetables and fruits. PMID:23443396
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Cramon, Per; Winther, Kristian Hillert; Watt, Torquil; Bonnema, Steen Joop; Bjorner, Jakob Bue; Ekholm, Ola; Groenvold, Mogens; Hegedüs, Laszlo; Feldt-Rasmussen, Ulla; Rasmussen, Åse Krogh
2016-08-01
The treatment of hyperthyroidism is aimed at improving health-related quality of life (HRQoL) and reducing morbidity and mortality. However, few studies have used validated questionnaires to assess HRQoL prospectively in such patients. The purpose of this study was to assess the impact of hyperthyroidism and its treatment on HRQoL using validated disease-specific and generic questionnaires. This prospective cohort study enrolled 88 patients with Graves' hyperthyroidism and 68 with toxic nodular goiter from endocrine outpatient clinics at two Danish university hospitals. The patients were treated with antithyroid drugs, radioactive iodine, or surgery. Disease-specific and generic HRQoL were assessed using the thyroid-related patient-reported outcome (ThyPRO) and the Medical Outcomes Study 36-item Short Form (SF-36), respectively, evaluated at baseline and six-month follow-up. The scores were compared with those from two general population samples who completed ThyPRO (n = 739) and SF-36 (n = 6638). Baseline scores for patients with Graves' hyperthyroidism and toxic nodular goiter were significantly worse than those for the general population scores on all comparable ThyPRO scales and all SF-36 scales and component summaries. ThyPRO scores improved significantly with treatment on all scales in Graves' hyperthyroidism and four scales in toxic nodular goiter, while SF-36 scores improved on five scales and both component summaries in Graves' hyperthyroidism and only one scale in toxic nodular goiter. In Graves' hyperthyroidism, large treatment effects were observed on three ThyPRO scales (Hyperthyroid Symptoms, Tiredness, Overall HRQoL) and moderate effects on three scales (Anxiety, Emotional Susceptibility, Impaired Daily Life), while moderate effects were seen in two ThyPRO scales in toxic nodular goiter (Anxiety, Overall HRQoL). However, significant disease-specific and generic HRQoL deficits persisted on multiple domains across both patient groups. Graves' hyperthyroidism and toxic nodular goiter cause severe disease-specific and generic HRQoL impairments, and HRQoL deficits persist in both patient groups six months after treatment. These data have the potential to improve communication between physicians and patients by offering realistic estimates of expected HRQoL impairments and treatment effects. Future studies should identify risk factors for persistent HRQoL deficits, compare HRQoL effects of the various therapies, and thereby aid in determining the optimal treatment strategies.
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NASA Astrophysics Data System (ADS)
Allegra Mascaro, Anna Letizia; Costantini, Irene; Margoni, Emilia; Iannello, Giulio; Bria, Alessandro; Sacconi, Leonardo; Pavone, Francesco S.
2016-03-01
Two-photon imaging combined with targeted fluorescent indicators is extensively used for visualizing critical features of brain functionality and structural plasticity. Back-scattered photons from the NIR laser provide complimentary information without introducing any exogenous labelling. Here, we describe a versatile approach that, by collecting the reflected NIR light, provides structural details on the myelinated axons and blood vessels in the brain, both in fixed samples and in live animals. Indeed, by combining NIR reflectance and two-photon imaging of a slice of hippocampus from Thy1-GFPm mice, we show the presence of randomly oriented axons intermingled with sparsely fluorescent neuronal processes. The back-scattered photons guide the contextualization of the fluorescence structure within brain atlas thanks to the recognition of characteristic hippocampal structures. Label-free detection of axonal elongations over the layer 2/3 of mouse cortex under a cranial window was also possible in live brain. Finally, blood flow could be measured in vivo, thus validating label free NIR reflectance as a tool for monitoring hemodynamic fluctuations. The prospective versatility of this label-free technique complimentary to two-photon fluorescence microscopy is demonstrated in a mouse model of photothrombotic stroke in which the axonal degeneration and blood flow remodeling can be investigated simultaneously.
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U.S. EPA, Pesticide Product Label, FOOD PLANT FOGGING INSECTICIDE, 01/30/1990
2011-04-14
... w.lter. food, 01" fet:,1 hy ')lur.J,:!C 01 ,li· ••• u"> •• I. ... (his proJuct c. .• Z",.. IfitJ~f'!f.1.~. I)n thi') label" •• o 6""(l'r .' ... IJlIIC'i ,111 ri'1lks of J\\t~ ')lo.SI~I~ or'. ...
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[Distal hereditary motor neuropathy].
Devic, P; Petiot, P
2011-11-01
Distal hereditary motor neuropathy (dHMN), also known as spinal muscular atrophy, represents a group of clinically and genetically heterogeneous diseases caused by degenerations of spinal motor neurons and leading to distal muscle weakness and wasting. Nerve conduction studies reveal a pure motor axonopathy and needle examination shows chronic denervation. dHMN were initially subdivided into seven subtypes according to mode of inheritance, age at onset, and clinical evolution. Recent studies have shown that these subtypes are still heterogeneous at the molecular genetic level and novel clinical and genetic entities have been characterized. To date, mutations in 11 different genes have been identified for autosomal-dominant, autosomal-recessive, and X-linked recessive dHMN. Most of the genes encode protein involved in housekeeping functions, endosomal trafficking, axonal transport, translation synthesis, RNA processing, oxidative stress response and apoptosis. The pathophysiological mechanisms underlying dHMN seem to be related to the "length-dependent" death of motor neurons of the anterior horn of the spinal cord, likely because their large axons have higher metabolic requirements for maintenance. dHMN remain heterogeneous at the clinical and molecular genetic level. The molecular pathomechanisms explaining why mutations in these ubiquitously expressed housekeeping genes result in the selective involvement of spinal motor neurons remain to be unravelled. Copyright © 2011 Elsevier Masson SAS. All rights reserved.
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Interactions between β-amyloid and central cholinergic neurons: implications for Alzheimer's disease
Kar, Satyabrata; Slowikowski, Stephen P.M.; Westaway, David; Mount, Howard T.J.
2004-01-01
Alzheimer's disease is an age-related neurodegenerative disorder that is characterized by a progressive loss of memory and deterioration of higher cognitive functions. The brain of an individual with Alzheimer's disease exhibits extracellular plaques of aggregated β-amyloid protein (Aβ), intracellular neurofibrillary tangles that contain hyperphosphorylated tau protein and a profound loss of basal forebrain cholinergic neurons that innervate the hippocampus and the neocortex. Aβ accumulation may trigger or contribute to the process of neurodegeneration. However, the mechanisms whereby Aβ induces basal forebrain cholinergic cell loss and cognitive impairment remain obscure. Physiologically relevant concentrations of Aβ-related peptides have acute, negative effects on multiple aspects of acetylcholine (ACh) synthesis and release, without inducing toxicity. These data suggest a neuromodulatory influence of the peptides on central cholinergic functions. Long-term exposure to micromolar Aβ induces cholinergic cell toxicity, possibly via hyperphosphorylation of tau protein. Conversely, activation of selected cholinergic receptors has been shown to alter the processing of the amyloid precursor protein as well as phosphorylation of tau protein. A direct interaction between Aβ and nicotinic ACh receptors has also been demonstrated. This review addresses the role of Aβ-related peptides in regulating the function and survival of central cholinergic neurons and the relevance of these effects to cholinergic deficits in Alzheimer's disease. Understanding the functional interrelations between Aβ peptides, cholinergic neurons and tau phosphorylation will unravel the biologic events that precede neurodegeneration and may lead to the development of more effective pharmacotherapies for Alzheimer's disease. PMID:15644984
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Narayanan, Rajeevan T.; Egger, Robert; Johnson, Andrew S.; Mansvelder, Huibert D.; Sakmann, Bert; de Kock, Christiaan P.J.; Oberlaender, Marcel
2015-01-01
Vertical thalamocortical afferents give rise to the elementary functional units of sensory cortex, cortical columns. Principles that underlie communication between columns remain however unknown. Here we unravel these by reconstructing in vivo-labeled neurons from all excitatory cell types in the vibrissal part of rat primary somatosensory cortex (vS1). Integrating the morphologies into an exact 3D model of vS1 revealed that the majority of intracortical (IC) axons project far beyond the borders of the principal column. We defined the corresponding innervation volume as the IC-unit. Deconstructing this structural cortical unit into its cell type-specific components, we found asymmetric projections that innervate columns of either the same whisker row or arc, and which subdivide vS1 into 2 orthogonal [supra-]granular and infragranular strata. We show that such organization could be most effective for encoding multi whisker inputs. Communication between columns is thus organized by multiple highly specific horizontal projection patterns, rendering IC-units as the primary structural entities for processing complex sensory stimuli. PMID:25838038
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Umapathy, Nagavedi S; Dun, Ying; Martin, Pamela M; Duplantier, Jennifer N; Roon, Penny; Prasad, Puttur; Smith, Sylvia B; Ganapathy, Vadivel
2008-11-01
Glutamine transport is essential for the glutamate-glutamine cycle, which occurs between neurons and glia. System N, consisting of SN1 (SNAT3) and SN2 (SNAT5), is the principal mediator of glutamine transport in retinal Müller cells. Mediators of glutamine transport in retinal ganglion cells were investigated. The relative contributions of various transport systems for glutamine uptake (systems N, A, L, y+L, ASCT, and ATB(0,+)) were examined in RGC-5 cells based on differential features of the individual transport systems. mRNA for the genes encoding members of these transport systems were analyzed by RT-PCR. Based on these data, SN1 and SN2 were analyzed in mouse retina, RGC-5 cells, and primary mouse ganglion cells (GCs) by in situ hybridization (ISH), immunofluorescence (IF), and Western blotting. Three transport systems--N, A, and L--participated in glutamine uptake in RGC-5 cells. System N was the principal contributor; systems A and L contributed considerably less. ISH and IF revealed SN1 and SN2 expression in the ganglion, inner nuclear, and photoreceptor cell layers. SN1 and SN2 colocalized with the ganglion cell marker Thy 1.2 and with the Müller cell marker vimentin, confirming their presence in both retinal cell types. SN1 and SN2 proteins were detected in primary mouse GCs. These findings suggest that in addition to its role in glutamine uptake in retinal glial cells, system N contributes significantly to glutamine uptake in ganglion cells and, hence, contributes to the retinal glutamate-glutamine cycle.
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NASA Astrophysics Data System (ADS)
Gabrieli, Andrea; Wright, Robert; Lucey, Paul G.; Porter, John N.; Garbeil, Harold; Pilger, Eric; Wood, Mark
2016-10-01
The ability to image and quantify SO2 path-concentrations in volcanic plumes, either by day or by night, is beneficial to volcanologists. Gas emission rates are affected by the chemical equilibria in rising magmas and a better understanding of this relationship would be useful for short-term eruption prediction. A newly developed remote sensing long-wave thermal InfraRed (IR) imaging hyperspectral sensor - the Thermal Hyperspectral Imager (THI) - was built and tested. The system employs a Sagnac interferometer and an uncooled microbolometer in rapid scanning configuration to collect hyperspectral images of volcanic plumes. Each pixel in the resulting image yields a spectrum with 50 samples between 8 and 14 μm. Images are spectrally and radiometrically calibrated using an IR source with a narrow band filter and two blackbodies. In this paper, the sensitivity of the instrument for the purpose of quantifying SO2 using well constrained laboratory experiments is evaluated, and initial field results from Kīlauea volcano, Hawai'i, are presented. The sensitivity of THI was determined using gas cells filled with known concentrations of SO2 and using NIST-traceable blackbodies to simulate a range of realistic background conditions. Measurements made by THI were then benchmarked against a high spectral resolution off-the-shelf Michelson FTIR instrument. Theoretical thermal IR spectral radiances were computed with MODTRAN5 for the same optical conditions, to evaluate how well the (known) concentration of SO2 in the gas cells could be retrieved from the resulting THI spectra. Finally, THI was recently field-tested at Kīlauea to evaluate its ability to image the concentration of SO2 in a real volcanic plume. A path-concentration of 7150 ppm m was retrieved from measurements made near the Halema'uma'u vent.
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NASA Astrophysics Data System (ADS)
Minka, N. S.; Ayo, J. O.
2012-03-01
The major factor in the induction of physiological stress during road transportation of livestock is the complex fluctuations of the thermal transport microenvironment, encountered when animals are transported across different ecological zones. Recommended guidelines on optimum "on-board" conditions in which goats should be transported are lacking, and there are no acceptable ranges and limits for the thermal loads to which goats may be subjected during long-distance road transportation in hot-dry conditions. Panting score (PS), rectal temperature (RT), heart rate (HR) and respiratory rate (RR) were employed as reliable stress indices to assess the effects of different thermal loads, measured as temperature humidity index (THI), encountered in the vehicle during 12 h of road transportation of 40 goats, and to suggest the administration of 100 mg/kg body weight of ascorbic acid (AA) as an ameliorating agent. The results obtained showed that the PS, RT, HR and RR rose above normal reference values with increase in the THI and journey duration. The rise in PS value, which is a visual indicator of the severity of thermal load, was the most pronounced. The results suggest that values of THI in the vehicle up to 94.6 constitute no risk, while at of 100 it presents a moderate risk and above 100 may result in severe stress. The relationships between the thermal load and the physiological variables were positive and significant ( P < 0.05). They reflect the degree of stress imposed by each THI value during the transportation, and may be used as recommended ranges and limit thermal load values in transported goats. The results demonstrated that administration of 100 mg/kg body weight of AA before road transportation mitigated the risk of adverse effects of high THI values and other stress factors due to road transportation in goats.
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ACTIVE SUPPRESSION OF IMMUNOGLOBULIN ALLOTYPE SYNTHESIS
Herzenberg, Leonore A.; Chan, Eva L.; Ravitch, Myrnice M.; Riblet, Roy J.; Herzenberg, Leonard A.
1973-01-01
Thymus-derived cells (T cells) that actively suppress production of IgG2a immunoglobulins carrying the Ig-1b allotype have been found in adult (SJL x BALB/c)F1 mice exposed to anti-Ig-1b early in life. The suppression is specific for Ig-1b. The allelic product, Ig-1a, is unaffected. Spleen, lymph node, bone marrow, or thymus cells from suppressed mice suppress production of Ig-1b by syngeneic spleen cells from normal F1 mice. When a mixture of suppressed and normal cells is transferred into lethally irradiated BALB/c mice, there is a short burst of Ig-1b production after which Ig-1b levels in the recipient fall rapidly below detectability. Pretreatment of the cells from the suppressed mice with antiserum specific for T cells (anti-Thy-1b) plus complement before mixture destroys the suppressing activity. Similar results with suppressor cells were obtained in vitro using Mishell-Dutton cultures. Mixture of spleen cells from suppressed animals with sheep erythrocyte (SRBC)-primed syngeneic normal spleen before culture suppresses Ig-1b plaque-forming cell (PFC) formation while leaving Ig-1a PFC unaffected. Treatment of the suppressed spleen with anti-Thy-1b before transfer removes the suppressing activity. PMID:4541122
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NASA Astrophysics Data System (ADS)
Stepanova, N. V.; Arkhipova, N. S.; Fomina, S. F.
2018-01-01
Assessment of non-carcinogenic risks from chemical substances ingested with drinking water included peroral, skin and inhalation routes of contact with water. The study was carried out for children aged 3-6 years living in 4 districts (zones) of the city of Kazan. Regional exposure factors (REF) at the median (Me) and the 95-th Percentile (95P) levels were identified according to the results of the questionnaire survey. The value of total hazard indices (THI) calculated with application of REF at the median (Me) and the 95-th Percentile (95P) levels made THIMe = 14.2 and 17.1, and THI 95perc = 13.03 and 16.3 in zones with a combined type of water supply. The ingestion of chemical substances with drinking water in different zones of the city of Kazan implies, alert and high levels of non-carcinogenic health risk for the child population.
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Sajad, Mir; Zargan, Jamil; Zargar, Mohammad Afzal; Sharma, Jyoti; Umar, Sadiq; Arora, Rajesh; Khan, Haider A
2013-05-01
Survival along with optimal proliferation of neuronal precursors determines the outcomes of the endogenous cellular repair in CNS. Cellular-oxidation based cell death has been described in several neurodegenerative disorders. Therefore, this study was aimed at the identification of the potent targets of oxidative damage to the neuronal precursors and its effective prevention by a natural flavonoid, Quercetin. Neuronal precursor cells (NPCs), Nestin+ and GFAP (Glial fibrillary acidic protein)+ were isolated and cultured from adult rat SVZ (subventricular zone). These cells were challenged with a single dose of H2O2 (50μM) and/or pre-treated with different concentrations of Quercetin. H2O2 severely limited the cellular viability and expansion of the neurospheres. Cellular-oxidation studies revealed reduction in glutathione dependent redox buffering along with depletion of enzymatic cellular antioxidants that might potentiate the nitrite (NO2(-)) and superoxide anion (O2(-)) mediated peroxynitrite (ONOO(-)) formation and irreversible protein nitration. We identified depleted PK-M2 (M2 isoform of pyruvate kinase) activity and apoptosis of NPCs revealed by the genomic DNA fragmentation and elevated PARP (poly ADP ribose polymerase) activity along with increased Caspase activity initiated by severely depolarised mitochondrial membranes. However, the pre-treatment of Quercetin in a dose-response manner prevented these changes and restored the expansion of neurospheres preferably by neutralizing the oxidative conditions and thereby reducing peroxynitrite formation, protein nitration and PK-M2 depletion. Our results unravel the potential interactions of oxidative environment and respiration in the survival and activation of precursors and offer a promise shown by a natural flavonoid in the protective strategy for neuronal precursors of adult brain. Copyright © 2013 Elsevier Inc. All rights reserved.
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Immuno-PCR assay for sensitive detection of proteins in real time
USDA-ARS?s Scientific Manuscript database
The immuno-PCR (IPCR) assay combines the versatility and robustness of immunoassays with the exponential signal amplification power of the polymerase chain reaction (PCR). Typically, IPCR allows a 10–1,000-fold increase in sensitivity over the analogous enzyme-linked immunosorbent assay (ELISA). Thi...
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A Humanized Mouse Model Generated Using Surplus Neonatal Tissue.
Brown, Matthew E; Zhou, Ying; McIntosh, Brian E; Norman, Ian G; Lou, Hannah E; Biermann, Mitch; Sullivan, Jeremy A; Kamp, Timothy J; Thomson, James A; Anagnostopoulos, Petros V; Burlingham, William J
2018-04-10
Here, we describe the NeoThy humanized mouse model created using non-fetal human tissue sources, cryopreserved neonatal thymus and umbilical cord blood hematopoietic stem cells (HSCs). Conventional humanized mouse models are made by engrafting human fetal thymus and HSCs into immunocompromised mice. These mice harbor functional human T cells that have matured in the presence of human self-peptides and human leukocyte antigen molecules. Neonatal thymus tissue is more abundant and developmentally mature and allows for creation of up to ∼50-fold more mice per donor compared with fetal tissue models. The NeoThy has equivalent frequencies of engrafted human immune cells compared with fetal tissue humanized mice and exhibits T cell function in assays of ex vivo cell proliferation, interferon γ secretion, and in vivo graft infiltration. The NeoThy model may provide significant advantages for induced pluripotent stem cell immunogenicity studies, while bypassing the requirement for fetal tissue. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.
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Yang, Qi; Ge, Moyar Q.; Kokalari, Blerina; Redai, Imre G.; Wang, Xinxin; Kemeny, David M.; Bhandoola, Avinash; Haczku, Angela
2015-01-01
Background Patients with asthma are highly susceptible to air pollution and in particular, to the effects of ozone (O3) inhalation, but the underlying mechanisms remain unclear. Objective Using mouse models of O3-induced airway inflammation and hyperresponsiveness (AHR), we sought to investigate the role of the recently discovered group 2 innate lymphoid cells (ILC2). Methods C57BL/6 and Balb/c mice were exposed to Aspergillus fumigatus and/or O3 (2ppm, 2h). ILC2 were isolated by FACS sorting and studied for IL-5 and IL-13 mRNA expression. ILC2 were depleted with anti-Thy1.2 mAb and replaced by intratracheal transfer of ex vivo expanded Thy1.1 ILC2. Cytokines (ELISA, qPCR), inflammatory cell profile and AHR (FlexiVent) were assessed in the mice. Results In addition to neutrophil influx, O3 inhalation elicited the appearance of eosinophils and IL-5 in the airways of Balb/c but not C57BL/6 mice. Although O3 induced expression of IL-33, a known activator of ILC2 in the lung was similar between these strains, isolated pulmonary ILC2 from O3 exposed Balb/c mice had significantly greater IL-5 and IL-13 mRNA expression than those of C57BL/6 mice. This suggested that an altered ILC2 function in Balb/c mice may mediate the increased O3 responsiveness. Indeed, anti-Thy1.2 treatment abolished, whereas ILC2 add-back dramatically enhanced O3-induced AHR. Conclusions O3-induced activation of pulmonary ILC2 was necessary and sufficient to mediate asthma-like changes in Balb/c mice. This previously unrecognized role of ILC2 may help explain the heightened susceptibility of human asthmatic airways to O3 exposure. PMID:26282284
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NASA Astrophysics Data System (ADS)
Battini, Monica; Barbieri, Sara; Fioni, Luna; Mattiello, Silvana
2016-02-01
This investigation tested the feasibility and validity of indicators of cold and heat stress in dairy goats for on-farm welfare assessment protocols. The study was performed on two intensive dairy farms in Italy. Two different 3-point scale (0-2) scoring systems were applied to assess cold and heat stress. Cold and heat stress scores were visually assessed from outside the pen in the morning, afternoon and evening in January-February, April-May and July 2013 for a total of nine sessions of observations/farm. Temperature (°C), relative humidity (%) and wind speed (km/h) were recorded and Thermal Heat Index (THI) was calculated. The sessions were allocated to three climatic seasons, depending on THI ranges: cold (<50), neutral (50-65) and hot (>65). Score 2 was rarely assessed; therefore, scores 1 and 2 were aggregated for statistical analysis. The amount of goats suffering from cold stress was significantly higher in the cold season than in neutral ( P < 0.01) and hot ( P < 0.001) seasons. Signs of heat stress were recorded only in the hot season ( P < 0.001). The visual assessment from outside the pen confirms the on-farm feasibility of both indicators: No constraint was found and time required was less than 10 min. Our results show that cold and heat stress scores are valid indicators to detect thermal stress in intensively managed dairy goats. The use of a binary scoring system (presence/absence), merging scores 1 and 2, may be a further refinement to improve the feasibility. This study also allows the prediction of optimal ranges of THI for dairy goat breeds in intensive husbandry systems, setting a comfort zone included into 55 and 70.
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Divergent mechanisms of insulin-like growth factor I and II on rat hepatocyte proliferation.
Raper, S; Kothary, P; Ishoo, E; Dikin, M; Kokudo, N; Hashimoto, M; DeMatteo, R P
1995-07-21
Insulin-like growth factors I and II are peptides with a structural homology for proinsulin, and are involved in hepatocyte proliferation. IGF-I and IGF-II, however, have different metabolic roles, and their mechanisms of action are incompletely known. We hypothesized that IGF-I and IGF-II act by different signal transduction pathways. To test this hypothesis, hepatocytes from 200 g male Sprague-Dawley rats were isolated by a two-step collagenase perfusion technique and plated at a density of 10(5) cells/16 mm Primaria plate. Proliferation was measured by [3H]thymidine ([3H]thy) incorporation into DNA, and an autoradiographic nuclear labeling index (LI). To analyze signal transduction, cyclic AMP (cAMP) levels were measured 5 min after addition of reagents by a radioimmunoassay. Reagents (doses) used were: IGF-I (2 nM), IGF-II (2 nM), the inhibitory peptide somatostatin-14 (SS14) (10 nM), and the adenylyl cyclase antagonist dideoxyadenosine (DDA) (10 microM). A summary of the findings is as follows: (1) IGF-I stimulates [3H]thy, LI and cAMP accumulation. (2) IGF-II stimulates [3H]thy and LI but not cAMP; (3) IGF-I but not IGF-II effects are inhibited by SS14 and DDA. We conclude that the hepatotrophic effects of IGF-I and IGF-II occur by different mechanisms: IGF-I is cAMP-dependent, IGF-II is cAMP-independent.
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Watt, Torquil; Groenvold, Mogens; Hegedüs, Laszlo; Bonnema, Steen Joop; Rasmussen, Åse Krogh; Feldt-Rasmussen, Ulla; Bjorner, Jakob Bue
2014-02-01
To evaluate the extent of differential item functioning (DIF) within the thyroid-specific quality of life patient-reported outcome measure, ThyPRO, according to sex, age, education and thyroid diagnosis. A total of 838 patients with benign thyroid diseases completed the ThyPRO questionnaire (84 five-point items, 13 scales). Uniform and nonuniform DIF were investigated using ordinal logistic regression, testing for both statistical significance and magnitude (∆R(2) > 0.02). Scale level was estimated by the sum score, after purification. Twenty instances of DIF in 17 of the 84 items were found. Eight according to diagnosis, where the goiter scale was the one most affected, possibly due to differing perceptions in patients with auto-immune thyroid diseases compared to patients with simple goiter. Eight DIFs according to age were found, of which 5 were in positively worded items, which younger patients were more likely to endorse; one according to gender: women were more likely to report crying, and three according to educational level. The vast majority of DIF had only minor influence on the scale scores (0.1-2.3 points on the 0-100 scales), but two DIF corresponded to a difference of 4.6 and 9.8, respectively. Ordinal logistic regression identified DIF in 17 of 84 items. The potential impact of this on the present scales was low, but items displaying DIF could be avoided when developing abbreviated scales, where the potential impact of DIF (due to fewer items) will be larger.
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Capitani, Paolo; Cerri, Matteo; Amici, Roberto; Baracchi, Francesca; Jones, Christine Ann; Luppi, Marco; Perez, Emanuele; Parmeggiani, Pier Luigi; Zamboni, Giovanni
A shift of physiological regulations from a homeostatic to a non-homeostatic modality characterizes the passage from non-NREM sleep (NREMS) to REM sleep (REMS). In the rat, an EEG index which allows the automatic scoring of transitions from NREMS to REMS has been proposed: the NREMS to REMS transition indicator value, NIV [J.H. Benington et al., Sleep 17 (1994) 28-36]. However, such transitions are not always followed by a REMS episode, but are often followed by an awakening. In the present study, the relationship between changes in EEG activity and hypothalamic temperature (Thy), taken as an index of autonomic activity, was studied within a window consisting of the 60s which precedes a state change from a consolidated NREMS episode. Furthermore, the probability that a transition would lead to REMS or wake was analysed. The results showed that, within this time window, both a modified NIV (NIV(60)) and the difference between Thy at the limits of the window (Thy(D)) were related to the probability of REMS onset. Both the relationship between the indices and the probability of REMS onset was sigmoid, the latter of which saturated at a probability level around 50-60%. The efficacy for the prediction of successful transitions from NREMS to REMS found using Thy(D) as an index supports the view that such a transition is a dynamic process where the physiological risk to enter REMS is weighted at a central level.
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1983-04-01
Honorary Fraternity Gamma Eta Chapter- Secretary A-12 JAMES F. BRAITHWAITE, P.E. QUALIFICATIONS SUIMIARY PRINCIPAL James Braithwaite is Executive Vice...X factor Score subtotaLd/aaius .Me aubtOtal .7 C. oigmast pathway mabe "cre. l t~ thi ghest sabfoe Valueoo a. S-1. 9 -2 at 0-3 aboe. pathaysSubeorv
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Terashima, Hideki; Kato, Mikio; Ebisawa, Masayuki; Kobayashi, Hideki; Suzuki, Kanae; Nezu, Yoshikazu; Sada, Toshio
2014-07-05
R-268712 is a novel and specific inhibitor of activin receptor-like kinase 5 (ALK5), a transforming growth factor β (TGF-β) type I receptor. Evaluation of in vitro inhibition indicated that R-268712 is a potent and selective inhibitor of ALK5 with an IC50 of 2.5nM, an approximately 5000-fold more selectivity for ALK5 than p38 mitogen-activated protein kinase (MAPK). Oral administration of R-268712 at doses of 1, 3 and 10mg/kg also inhibited the development of renal fibrosis in a dose-dependent manner in a unilateral ureteral obstruction (UUO) model. Additionally, we evaluated the efficacy of R-268712 in a heminephrectomized anti-Thy1 glomerulonephritis model at doses of 0.3 and 1mg/kg. R-268712 reduced proteinuria and glomerulosclerosis significantly with improvement of renal function. Collectively, these results suggested that R-268712 and other ALK5 inhibitors could suppress glomerulonephritis as well as glomerulosclerosis by an inhibitory mechanism that involves suppression of TGF-β signaling. Copyright © 2014 Elsevier B.V. All rights reserved.
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Suppression of Tinnitus in Chinese Patients Receiving Regular Cochlear Implant Programming.
Liu, Ying; Wang, Hong; Han, Dong Xu; Li, Ming Hua; Wang, Yu; Xiao, Yu Li
2016-04-01
To assess the clinical effect of cochlear implant programming on tinnitus. Tinnitus patients (n = 234) were divided into 3 groups: (1) preoperative tinnitus (n = 108), (2) postoperative tinnitus occurring before implant switch-on at week 4 (n = 88), and (3) tinnitus occurring more than 1 year postoperatively (n = 44). Patients in each group were randomly allocated into a programming subgroup that received programming for 12 weeks postoperatively or after tinnitus occurrence or a control subgroup. Impedance testing and the Tinnitus Handicap Inventory (THI) were performed preoperatively and at 4, 6, 8, and 12 weeks postoperatively (groups 1 and 2) or after tinnitus occurrence (group 3). Comparisons were performed using t tests and chi-square tests. Impedance was significantly lower in the programming subgroup than in the control subgroup in groups 1 and 2 at 8 and 12 weeks and in group 3 at 12 weeks. The THI scores decreased in both programming and control subgroups in all groups. However, this decrease was pronounced in the programming subgroup, whereas in the control subgroup, it occurred slowly over time. Cochlear implant programming decreases impedance and improves tinnitus symptoms. © The Author(s) 2015.
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Hagenmaier, J A; Reinhardt, C D; Bartle, S J; Thomson, D U
2016-12-01
Feedlot cattle ( = 1,395; BW = 568 ± 43 kg) were used to evaluate the effects of shade on animal welfare, growth performance, and carcass quality during the summer of 2013 in a Kansas commercial feedlot. Seven lots of predominately black steers and heifers (4 and 3, respectively) visually determined to be approaching the final mo on feed were identified, randomly gate-sorted, and allocated to pens located across the feed alley from each other to receive 1 of 2 treatments: 1) Shade (mean shade area = 1.5 m/ animal) or 2) No shade. Shade was provided using a 13-ounce polyethylene fabric and pens were oriented northwest to southeast. The mean starting date was June 13 and the mean days on feed for lots while on the study was 38 d. Cattle were fed a 77.67% DM steam-flaked corn-based diet and had ad libitum access to water throughout the duration of the trial. Zilpaterol hydrochloride (ZIL) was included in the finishing ration at an inclusion rate of 8.3 mg/kg of DM for the last 20 d on feed with a 3 d withdrawal period. Pen floor temperatures (PFT) were measured using an infrared thermometer and prevalence of cattle open-mouth breathing (OMB) was recorded on a pen basis. In addition to shade treatment, the effect of temperature humidity index (THI) on PFT and OMB was analyzed by classifying days as either "Alert" (THI < 79) or "Danger" (THI > 79). On the day of slaughter, pens within a replicate were kept separate through all stages of the marketing channel from loading at the feedlot until stunning at the plant. Pen served as the experimental unit for all measurements. There was a THI × shade treatment interaction for PFT and OMB ( < 0.001) where days classified as "Danger" increased PFT and prevalence of OMB compared to "Alert" days in unshaded but not shaded cattle. Shaded cattle had greater DMI ( = 0.01); however, unshaded cattle had greater G:F ( = 0.05) and therefore no differences were observed in ADG ( = 0.39). Shaded cattle had greater dressing percentage ( = 0.01), although HCW, LM area, fat thickness, marbling score, and quality grade did not differ between treatments ( > 0.05). Heat stress, a significant animal welfare concern and cause of reduced performance in feedlot cattle during the final phase of the feeding period, was alleviated in shaded cattle and illustrates the importance of shade provision as 1 tool to protect the welfare and increase feed consumption in large pens of feedlot cattle during hot summer months.
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MicroRNA let-7d regulates the TLX/microRNA-9 cascade to control neural cell fate and neurogenesis
Zhao, Chunnian; Sun, GuoQiang; Ye, Peng; Li, Shengxiu; Shi, Yanhong
2013-01-01
MicroRNAs have important functions in the nervous system through post-transcriptional regulation of neurogenesis genes. Here we show that microRNA let-7d, which has been implicated in cocaine addiction and other neurological disorders, targets the neural stem cell regulator TLX. Overexpression of let-7d in vivo reduced neural stem cell proliferation and promoted premature neuronal differentiation and migration, a phenotype similar to those induced by TLX knockdown or overexpression of its negatively-regulated target, microRNA-9. We found a let-7d binding sequence in the tlx 3′ UTR and demonstrated that let-7d reduced TLX expression levels in neural stem cells, which in turn, up-regulated miR-9 expression. Moreover, co-expression of let-7d and TLX lacking its 3′ UTR in vivo restored neural stem cell proliferation and reversed the premature neuronal differentiation and migration. Therefore, manipulating let-7d and its downstream targets could be a novel strategy to unravel neurogenic signaling pathways and identify potential interventions for relevant neurological disorders. PMID:23435502
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MicroRNA let-7d regulates the TLX/microRNA-9 cascade to control neural cell fate and neurogenesis.
Zhao, Chunnian; Sun, GuoQiang; Ye, Peng; Li, Shengxiu; Shi, Yanhong
2013-01-01
MicroRNAs have important functions in the nervous system through post-transcriptional regulation of neurogenesis genes. Here we show that microRNA let-7d, which has been implicated in cocaine addiction and other neurological disorders, targets the neural stem cell regulator TLX. Overexpression of let-7d in vivo reduced neural stem cell proliferation and promoted premature neuronal differentiation and migration, a phenotype similar to those induced by TLX knockdown or overexpression of its negatively-regulated target, microRNA-9. We found a let-7d binding sequence in the tlx 3' UTR and demonstrated that let-7d reduced TLX expression levels in neural stem cells, which in turn, up-regulated miR-9 expression. Moreover, co-expression of let-7d and TLX lacking its 3' UTR in vivo restored neural stem cell proliferation and reversed the premature neuronal differentiation and migration. Therefore, manipulating let-7d and its downstream targets could be a novel strategy to unravel neurogenic signaling pathways and identify potential interventions for relevant neurological disorders.
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The neural bases of host plant selection in a Neuroecology framework.
Reisenman, Carolina E; Riffell, Jeffrey A
2015-01-01
Understanding how animals make use of environmental information to guide behavior is a fundamental problem in the field of neuroscience. Similarly, the field of ecology seeks to understand the role of behavior in shaping interactions between organisms at various levels of organization, including population-, community- and even ecosystem-level scales. Together, the newly emerged field of "Neuroecology" seeks to unravel this fundamental question by studying both the function of neurons at many levels of the sensory pathway and the interactions between organisms and their natural environment. The interactions between herbivorous insects and their host plants are ideal examples of Neuroecology given the strong ecological and evolutionary forces and the underlying physiological and behavioral mechanisms that shaped these interactions. In this review we focus on an exemplary herbivorous insect within the Lepidoptera, the giant sphinx moth Manduca sexta, as much is known about the natural behaviors related to host plant selection and the involved neurons at several level of the sensory pathway. We also discuss how herbivore-induced plant odorants and secondary metabolites in floral nectar in turn can affect moth behavior, and the underlying neural mechanisms.
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Nociception and escape behavior in planarians
NASA Astrophysics Data System (ADS)
Schoetz Collins, Eva-Maria
2015-03-01
Planarians are famous and widely studied for their regenerative capabilities. When a moving planarian is cut through the middle, the resulting head and tail pieces instantaneously retract and exhibit a characteristic escape response that differs from normal locomotion. In asexual animals, a similar reaction is observed when the planarian undergoes fission, suggesting that reproduction through self-tearing is a rather traumatic event for the animal. Using a multiscale approach, we unravel the dynamics, mechanics, and functional aspects of the planarian escape response. This musculature-driven gait was found to be a dominating response that supersedes the urge to feed or reproduce and quantitatively differs from other modes of planarian locomotion (gliding, peristalsis). We show that this escape gait constitutes the animal's pain response mediated by TRP like receptors and the neurotransmitter histamine, and that it can be induced through adverse thermal, mechanical, electrical or chemical stimuli. Ultimately, we will examine the neuronal subpopulations involved in mediating escape reflexes in planarians and how they are functionally restored during regeneration, thereby gaining mechanistic insight into the neuronal circuits required for specific behaviors. Supported by BWF CASI and Sloan Foundation.
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USDA-ARS?s Scientific Manuscript database
Livestock play an important role in carbon cycling through consumption of biomass and emissions of methane. Recent research suggests that existing bottom-up inventories of livestock methane emissions in the U.S., such as those made using 2006 IPCC Tier 1 livestock emissions factors, are too low. Thi...
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Transfer of T-cell mediated immunity to Hymenolepis nana from mother mice to their neonates.
Asano, K; Okamoto, K
1992-01-15
Administration of lymph node cells from Hymenolepis nana-infected mice into lactating mothers, or directly suckling neonates successfully transferred immunity to the neonates. The capacity of lymph node cells to transfer immunity was completely abrogated by pretreatment with anti-Thy-1.2 monoclonal antibody and complement.
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Unravelling the NERDS syndrome.
Singh, Achintya Dinesh; Suri, Tejas Menon; Jagdish, Rakesh Kumar; Kumar, Uma
2018-05-12
A 22-year-old man presented with symmetric polyarthritis, pruritus and deviation of angle of mouth to the right side since the last 7 years. His symptoms were persistent despite receiving ayurvedic medications and symptomatic therapy. Examination revealed dry skin, cutaneous nodules, xanthelasma, periarticular non-tender swellings, pitting oedema of hands and feet and lower motor neuron type right facial palsy. Haematological investigations revealed eosinophilia and skin biopsy had cutaneous eosinophilic infiltration. The constellation of above findings comprises the nodules, eosinophilia, rheumatism, dermatitis and swelling syndrome. It a rare syndrome with few reported cases in literature. The patient was started on oral corticosteroids which was subsequently tapered and methotrexate therapy. His polyarthritis and skin rashes resolved with therapy. He has been followed-up for 2 years and is presently asymptomatic for the last 1 year. © BMJ Publishing Group Ltd (unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.
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Cole, Elizabeth; Brown, Traci A; Pinkerton, Kent E; Postma, Britten; Malany, Keegan; Yang, Mihi; Kim, Yang Jee; Hamilton, Raymond F; Holian, Andrij; Cho, Yoon Hee
2017-08-01
Prenatal and early-life environmental tobacco smoke (ETS) exposure can induce epigenetic alterations associated with inflammation and respiratory disease. The objective of this study was to address the long-term epigenetic consequences of perinatal ETS exposure on latent respiratory disease risk, which are still largely unknown. C57BL/6 mice were exposed to prenatal and early-life ETS; offspring lung pathology, global DNA, and gene-specific methylation were measured at two adult ages. Significant alterations in global DNA methylation and promoter methylation of IFN-γ and Thy-1 were found in ETS-exposed offspring at 10-12 and 20 weeks of age. These sustained epigenetic alterations preceded the onset of significant pulmonary pathologies observed at 20 weeks of age. This study suggests that perinatal ETS exposure induces persistent epigenetic alterations in global DNA, as well as IFN-γ and Thy-1 promoter methylation that precede the adult onset of fibrotic lung pathology. These epigenetic findings could represent potential biomarkers of latent respiratory disease risk.
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Euros, Pounds and Albion at Arms: European Monetary Policy and British Defence in the 21st Century
2004-09-01
Shakespeare , Hamlet , Prince of Denmark, act 1, scene 2. 2 William Shakespeare , Richard the Second, act 2, scene 1. 3 Hugo Young, This Blessed Plot: Britain...THIS PAGE INTENTIONALLY LEFT BLANK 1 I. INTRODUCTION Queen: Good Hamlet , cast thy nighted colour...tis common. All that lives must die, Passing through nature to eternity. Hamlet : Ay, madam, it is common. Queen: If it
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Coherent anti-Stokes Raman scattering enhancement of thymine adsorbed on graphene oxide
2014-01-01
Coherent anti-Stokes Raman scattering (CARS) of carbon nanostructures, namely, highly oriented pyrolytic graphite, graphene nanoplatelets, graphene oxide, and multiwall carbon nanotubes as well CARS spectra of thymine (Thy) molecules adsorbed on graphene oxide were studied. The spectra of the samples were compared with spontaneous Raman scattering (RS) spectra. The CARS spectra of Thy adsorbed on graphene oxide are characterized by shifts of the main bands in comparison with RS. The CARS spectra of the initial nanocarbons are definitely different: for all investigated materials, there is a redistribution of D- and G-mode intensities, significant shift of their frequencies (more than 20 cm-1), and appearance of new modes about 1,400 and 1,500 cm-1. The D band in CARS spectra is less changed than the G band; there is an absence of 2D-mode at 2,600 cm-1 for graphene and appearance of intensive modes of the second order between 2,400 and 3,000 cm-1. Multiphonon processes in graphene under many photon excitations seem to be responsible for the features of the CARS spectra. We found an enhancement of the CARS signal from thymine adsorbed on graphene oxide with maximum enhancement factor about 105. The probable mechanism of CARS enhancement is discussed. PMID:24948887
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Talking back: Development of the olivocochlear efferent system.
Frank, Michelle M; Goodrich, Lisa V
2018-06-26
Developing sensory systems must coordinate the growth of neural circuitry spanning from receptors in the peripheral nervous system (PNS) to multilayered networks within the central nervous system (CNS). This breadth presents particular challenges, as nascent processes must navigate across the CNS-PNS boundary and coalesce into a tightly intermingled wiring pattern, thereby enabling reliable integration from the PNS to the CNS and back. In the auditory system, feedforward spiral ganglion neurons (SGNs) from the periphery collect sound information via tonotopically organized connections in the cochlea and transmit this information to the brainstem for processing via the VIII cranial nerve. In turn, feedback olivocochlear neurons (OCNs) housed in the auditory brainstem send projections into the periphery, also through the VIII nerve. OCNs are motor neuron-like efferent cells that influence auditory processing within the cochlea and protect against noise damage in adult animals. These aligned feedforward and feedback systems develop in parallel, with SGN central axons reaching the developing auditory brainstem around the same time that the OCN axons extend out toward the developing inner ear. Recent findings have begun to unravel the genetic and molecular mechanisms that guide OCN development, from their origins in a generic pool of motor neuron precursors to their specialized roles as modulators of cochlear activity. One recurrent theme is the importance of efferent-afferent interactions, as afferent SGNs guide OCNs to their final locations within the sensory epithelium, and efferent OCNs shape the activity of the developing auditory system. This article is categorized under: Nervous System Development > Vertebrates: Regional Development. © 2018 Wiley Periodicals, Inc.
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The CB₁ cannabinoid receptor signals striatal neuroprotection via a PI3K/Akt/mTORC1/BDNF pathway.
Blázquez, C; Chiarlone, A; Bellocchio, L; Resel, E; Pruunsild, P; García-Rincón, D; Sendtner, M; Timmusk, T; Lutz, B; Galve-Roperh, I; Guzmán, M
2015-10-01
The CB1 cannabinoid receptor, the main molecular target of endocannabinoids and cannabis active components, is the most abundant G protein-coupled receptor in the mammalian brain. In particular, the CB1 receptor is highly expressed in the basal ganglia, mostly on terminals of medium-sized spiny neurons, where it plays a key neuromodulatory function. The CB1 receptor also confers neuroprotection in various experimental models of striatal damage. However, the assessment of the physiological relevance and therapeutic potential of the CB1 receptor in basal ganglia-related diseases is hampered, at least in part, by the lack of knowledge of the precise mechanism of CB1 receptor neuroprotective activity. Here, by using an array of pharmacological, genetic and pharmacogenetic (designer receptor exclusively activated by designer drug) approaches, we show that (1) CB1 receptor engagement protects striatal cells from excitotoxic death via the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin complex 1 pathway, which, in turn, (2) induces brain-derived neurotrophic factor (BDNF) expression through the selective activation of BDNF gene promoter IV, an effect that is mediated by multiple transcription factors. To assess the possible functional impact of the CB1/BDNF axis in a neurodegenerative-disease context in vivo, we conducted experiments in the R6/2 mouse, a well-established model of Huntington's disease, in which the CB1 receptor and BDNF are known to be severely downregulated in the dorsolateral striatum. Adeno-associated viral vector-enforced re-expression of the CB1 receptor in the dorsolateral striatum of R6/2 mice allowed the re-expression of BDNF and the concerted rescue of the neuropathological deficits in these animals. Collectively, these findings unravel a molecular link between CB1 receptor activation and BDNF expression, and support the relevance of the CB1/BDNF axis in promoting striatal neuron survival.
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Impact Damage Thresholds in Brittle Materials Impacted by Water Drops.
1979-03-01
hardness AS$TNAC T (C 5M.. cc roc .. aId. It ,. m.p .id td ..tIl~- S~. block cad ..) The damage threshold for brittle materials impacted by water drops...center, vls-a-vls their(continued) ~~~ DO I JA N71 1Q3 EDITION OP 1 NOV IS iS OHOLETE UNCLASSIFIED SECURITY CLASSIFICATION OP ThiS RAGS (a. ONs
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A Brain Unfixed: Unlimited Neurogenesis and Regeneration of the Adult Planarian Nervous System
Brown, David D. R.; Pearson, Bret J.
2017-01-01
Powerful genetic tools in classical laboratory models have been fundamental to our understanding of how stem cells give rise to complex neural tissues during embryonic development. In contrast, adult neurogenesis in our model systems, if present, is typically constrained to one or a few zones of the adult brain to produce a limited subset of neurons leading to the dogma that the brain is primarily fixed post-development. The freshwater planarian (flatworm) is an invertebrate model system that challenges this dogma. The planarian possesses a brain containing several thousand neurons with very high rates of cell turnover (homeostasis), which can also be fully regenerated de novo from injury in just 7 days. Both homeostasis and regeneration depend on the activity of a large population of adult stem cells, called neoblasts, throughout the planarian body. Thus, much effort has been put forth to understand how the flatworm can continually give rise to the diversity of cell types found in the adult brain. Here we focus on work using single-cell genomics and functional analyses to unravel the cellular hierarchies from stem cell to neuron. In addition, we will review what is known about how planarians utilize developmental signaling to maintain proper tissue patterning, homeostasis, and cell-type diversity in their brains. Together, planarians are a powerful emerging model system to study the dynamics of adult neurogenesis and regeneration. PMID:28588444
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Using induced pluripotent stem cells derived neurons to model brain diseases.
McKinney, Cindy E
2017-07-01
The ability to use induced pluripotent stem cells (iPSC) to model brain diseases is a powerful tool for unraveling mechanistic alterations in these disorders. Rodent models of brain diseases have spurred understanding of pathology but the concern arises that they may not recapitulate the full spectrum of neuron disruptions associated with human neuropathology. iPSC derived neurons, or other neural cell types, provide the ability to access pathology in cells derived directly from a patient's blood sample or skin biopsy where availability of brain tissue is limiting. Thus, utilization of iPSC to study brain diseases provides an unlimited resource for disease modelling but may also be used for drug screening for effective therapies and may potentially be used to regenerate aged or damaged cells in the future. Many brain diseases across the spectrum of neurodevelopment, neurodegenerative and neuropsychiatric are being approached by iPSC models. The goal of an iPSC based disease model is to identify a cellular phenotype that discriminates the disease-bearing cells from the control cells. In this mini-review, the importance of iPSC cell models validated for pluripotency, germline competency and function assessments is discussed. Selected examples for the variety of brain diseases that are being approached by iPSC technology to discover or establish the molecular basis of the neuropathology are discussed.
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2014-01-01
Background The aim of the present study was to investigate, which aspects of tinnitus are most relevant for impairment of quality of life. For this purpose we analysed how responses to the Tinnitus Handicap Inventory (THI) and to the question “How much of a problem is your tinnitus at present” correlate with the different aspects of quality of life and depression. Methods 1274 patients of the Tinnitus Research Initiative database were eligible for analysis. The Tinnitus Research Initiative database is composed of eight study centres from five countries. We assessed to which extent the Tinnitus Handicap Inventory (THI) and its subscales and single items as well as the tinnitus severity correlate with Beck Depression Inventory (BDI) score and different domains of the short version of the WHO-Quality of Life questionnaire (WHO-QoL Bref) by means of simple and multiple linear regression models. Results The THI explained considerable portions of the variance of the WHO-QoL Physical Health (R2 = 0.39) and Psychological Health (R2 = 0.40) and the BDI (R2 = 0.46). Furthermore, multiple linear regression models which included each THI item separately explained an additional 5% of the variance compared to the THI total score. The items feeling confused from tinnitus, the trouble of falling asleep at night, the interference with job or household responsibilities, getting upset from tinnitus, and the feeling of being depressed were those with the highest influence on quality of life and depression. The single question with regard to tinnitus severity explained 18%, 16%, and 20% of the variance of Physical Health, Psychological Health, and BDI respectively. Conclusions In the context of a cross-sectional correlation analysis, our findings confirmed the strong and consistent relationships between self-reported tinnitus burden and both quality of life, and depression. The single question “How much of a problem is your tinnitus” reflects tinnitus-related impairment in quality of life and can thus be recommended for use in clinical routine. PMID:24422941
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Zeman, Florian; Koller, Michael; Langguth, Berthold; Landgrebe, Michael
2014-01-14
The aim of the present study was to investigate, which aspects of tinnitus are most relevant for impairment of quality of life. For this purpose we analysed how responses to the Tinnitus Handicap Inventory (THI) and to the question "How much of a problem is your tinnitus at present" correlate with the different aspects of quality of life and depression. 1274 patients of the Tinnitus Research Initiative database were eligible for analysis. The Tinnitus Research Initiative database is composed of eight study centres from five countries. We assessed to which extent the Tinnitus Handicap Inventory (THI) and its subscales and single items as well as the tinnitus severity correlate with Beck Depression Inventory (BDI) score and different domains of the short version of the WHO-Quality of Life questionnaire (WHO-QoL Bref) by means of simple and multiple linear regression models. The THI explained considerable portions of the variance of the WHO-QoL Physical Health (R2 = 0.39) and Psychological Health (R2 = 0.40) and the BDI (R2 = 0.46). Furthermore, multiple linear regression models which included each THI item separately explained an additional 5% of the variance compared to the THI total score. The items feeling confused from tinnitus, the trouble of falling asleep at night, the interference with job or household responsibilities, getting upset from tinnitus, and the feeling of being depressed were those with the highest influence on quality of life and depression. The single question with regard to tinnitus severity explained 18%, 16%, and 20% of the variance of Physical Health, Psychological Health, and BDI respectively. In the context of a cross-sectional correlation analysis, our findings confirmed the strong and consistent relationships between self-reported tinnitus burden and both quality of life, and depression. The single question "How much of a problem is your tinnitus" reflects tinnitus-related impairment in quality of life and can thus be recommended for use in clinical routine.
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Tornuk, Fatih; Sagdic, Osman; Hancer, Mehmet; Yetim, Hasan
2018-05-01
In this study, a novel procedure was performed for grafting of nanoclays (montmorillonite (MMT) and halloysite (HNT)) with essential oil constituents (thymol (THY), eugenol (EUG) and carvacrol (CRV)) using Tween 80 as surfactant and then the nanoclay particles were incorporated into LLDPE pellets (5 wt%) to produce active nanocomposite films using a twin screw extruder. The resulting nanocomposite films were analyzed for antimicrobial and antioxidant capacity as well as thickness, mechanical, color, barrier, thermal properties and surface morphology and molecular composition. Release of the active compounds from the films at the refrigerated and room temperature conditions were also tested. The results showed that the films had strong in vitro antibacterial activity against pathogenic bacteria (Salmonella Typhimurium, Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus) while their effect against lactic acid bacteria (Lactobacillus rhamnosus and Lb. casei) was limited. The lowest and highest DPPH scavenging ability levels were 65.59% and % 87.92, belonged to THY-MMT and EUG-MMT, respectively. Release of active compounds at 24 °C was much more rapid than at 4 °C. CRV-HNT and THY-HNT provided slower release than the other films. SEM results showed that nanoclays were uniformly dispersed in the polymer matrix with exceptional agglomerates. Incorporation of the active nanoclays significantly (P > 0.05) improved tensile strength and elongation of the films. The results confirmed that LLDPE based active nanocomposite films could be successfully produced due to its good interaction with MMT and HNT, activated with THY, EUG and CRV. Copyright © 2018 Elsevier Ltd. All rights reserved.
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The impact of tinnitus on daily activities in adult tinnitus sufferers: A pilot study.
Moroe, Nomfundo F; Khoza-Shangase, Katijah
2014-08-27
Few South African studies have been published on the impact of tinnitus on quality of life of tinnitus sufferers, although evidence suggests that a large portion of the general population suffers from tinnitus. The current study aimed at describing the effects of tinnitus on the quality of life of the participants as measured by the Tinnitus Handicap Inventory (THI). In a cross-sectional descriptive study design, 27 participants took part in the study by completing a self-administered THI questionnaire and participating in a semi-structured interview. Descriptive and inferential statistics were used to analyse the data. Descriptively, content analysis was used to organise and convey results from the interviews. Participants reported a wide range of perceived disability on the THI. Results ranged from mild to catastrophic, with functional disability being most prominent in all participants, although there were differences when results were analysed according to gender. There was an association between gender and the type of perceived disability, although this was statistically non-significant (p > 0.05). Only 26% of the participants reported no effect on occupational performance and quality of life, with the remainder of the participants reporting a significant effect. Limited effective management strategies were reported to have been implemented - a significant implication for the audiologists. The results have implications for audiologists as they suggest that audiologists should take a detailed case history to determine the extent to which tinnitus affects the individual. Furthermore, audiologists should administer a scale such as the THI in the management of tinnitus.
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Analysis and explanation of the Thiéry-Wundt illusion.
Day, Ross H; Kimm, Andrew C
2010-01-01
The midpoint of the axis of bisection in a triangle appears to be displaced toward the apex so that the apical extent seems to be shorter than the basal extent, an effect originally reported in 1895 by Thiéry and later in 1898 by Wundt and, therefore, referred to here as the Thiéry-Wundt illusion. Following a demonstration strongly suggesting that the illusion is yet another version of the Müller-Lyer effect in a stimulus figure with inward-directed angles, four exploratory experiments designed to throw more light on this group of illusions are reported. The first showed that the effect occurs in an open angle, between converging lines that do not meet to form an apex, between converging stepped lines, and when one of the converging lines is removed, leaving a single oblique line. The other three experiments showed that the illusion also occurs in an open semicircle and a rectangular bracket, is weakened by the addition of a complete or partial baseline to form a triangle, and weakly but reliably when the angle is minimally formed from dots marking the ends of oblique lines. On the basis of these data, Judd's version of the conventional Müller-Lyer figure, and informal but easily repeatable observations, it is concluded that the Thiéry-Wundt illusion is, like other variants of the Müller-Lyer group of illusions, due to a process of directional biasing-an extension of the concept of biasing proposed originally by Morgan et al (1990, Vision Research 30 1793 1810).
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Hisada, Masayuki; Ota, Yoshihiro; Zhang, Xiuying; Cameron, Andrew M; Gao, Bin; Montgomery, Robert A; Williams, George Melville; Sun, Zhaoli
2015-01-01
Livers from Lewis rats fed with 7% alcohol for 5 weeks were used for transplantation. Reduced sized (50%) livers or whole livers were transplanted into normal DA recipients, which, in this strain combination, survive indefinitely when the donor has not been fed alcohol. However, none of the rats survived a whole fatty liver transplant while six of seven recipients of reduced sized alcoholic liver grafts survived long term. SDF-1 and HGF were significantly increased in reduced size liver grafts compared to whole liver grafts. Lineage-negative Thy-1+CXCR4+CD133+ stem cells were significantly increased in the peripheral blood and in allografts after reduced size fatty liver transplantation. In contrast, there were meager increases in cells reactive with anti Thy-1, CXCR4 and CD133 in peripheral blood and allografts in whole alcoholic liver recipients. The provision of plerixafor, a stem cell mobilizer, salvaged 5 of 10 whole fatty liver grafts. Conversely, blocking SDF-1 activity with neutralizing antibodies diminished stem cell recruitment and four of five reduced sized fatty liver recipients died. Thus chemokine insuficiency was associated with transplant failure of whole grafts which was overcome by the increased regenerative requirements promoted by the small grafts and mediated by SDF-1 resulting in stem cell influx. PMID:22994609
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Umapathy, Nagavedi S.; Dun, Ying; Martin, Pamela M.; Duplantier, Jennifer N.; Roon, Penny; Prasad, Puttur; Smith, Sylvia B.; Ganapathy, Vadivel
2008-01-01
Purpose Glutamine transport is essential for the glutamate-glutamine cycle, which occurs between neurons and glia. System N, consisting of SN1 (SNAT3) and SN2 (SNAT5), is the principal mediator of glutamine transport in retinal Müller cells. Mediators of glutamine transport in retinal ganglion cells were investigated. Methods The relative contributions of various transport systems for glutamine uptake (systems N, A, L, y+L, ASCT, and ATB0,+) were examined in RGC-5 cells based on differential features of the individual transport systems. mRNA for the genes encoding members of these transport systems were analyzed by RT-PCR. Based on these data, SN1 and SN2 were analyzed in mouse retina, RGC-5 cells, and primary mouse ganglion cells (GCs) by in situ hybridization (ISH), immunofluorescence (IF), and Western blotting. Results Three transport systems—N, A, and L—participated in glutamine uptake in RGC-5 cells. System N was the principal contributor; systems A and L contributed considerably less. ISH and IF revealed SN1 and SN2 expression in the ganglion, inner nuclear, and photoreceptor cell layers. SN1 and SN2 colocalized with the ganglion cell marker Thy 1.2 and with the Müller cell marker vimentin, confirming their presence in both retinal cell types. SN1 and SN2 proteins were detected in primary mouse GCs. Conclusions These findings suggest that in addition to its role in glutamine uptake in retinal glial cells, system N contributes significantly to glutamine uptake in ganglion cells and, hence, contributes to the retinal glutamate-glutamine cycle. PMID:18689705
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Kong, Fen-Ying; Xu, Mao-Tian; Xu, Jing-Juan; Chen, Hong-Yuan
2011-10-15
In this paper, gold nanoparticle-thionine-reduced graphene oxide (GNP-THi-GR) nanocomposites were prepared to design a label-free immunosensor for the sensitive detection of carcinoembryonic antigen (CEA). The nanocomposites with good biocompatibility, excellent redox electrochemical activity and large surface area were coated onto the glassy carbon electrode (GCE) surface and then CEA antibody (anti-CEA) was immobilized on the electrode to construct the immunosensor. The morphologies and electrochemistry of the formed nanocomposites were investigated by using scanning electron microscopy (SEM), ultraviolet-visible (UV-vis) spectrometry, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). CV and differential pulse voltammetry (DPV) studies demonstrated that the formation of antibody-antigen complexes decreased the peak current of THi in the GNP-THi-GR nanocomposites. The decreased currents were proportional to the CEA concentration in the range of 10-500 pg/mL with a detection limit of 4 pg/mL. The proposed method was simple, fast and inexpensive for the determination of CEA at very low levels. Copyright © 2011 Elsevier B.V. All rights reserved.
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Cerebellum: links between development, developmental disorders and motor learning
Manto, Mario U.; Jissendi, Patrice
2012-01-01
The study of the links and interactions between development and motor learning has noticeable implications for the understanding and management of neurodevelopmental disorders. This is particularly relevant for the cerebellum which is critical for sensorimotor learning. The olivocerebellar pathway is a key pathway contributing to learning of motor skills. Its developmental maturation and remodeling are being unraveled. Advances in genetics have led to major improvements in our appraisal of the genes involved in cerebellar development, especially studies in mutant mice. Cerebellar neurogenesis is compartmentalized in relationship with neurotransmitter fate. The Engrailed-2 gene is a major actor of the specification of cerebellar cell types and late embryogenic morphogenesis. Math1, expressed by the rhombic lip, is required for the genesis of glutamatergic neurons. Mutants deficient for the transcription factor Ptf1a display a lack of Purkinje cells and gabaergic interneurons. Rora gene contributes to the developmental signaling between granule cells and Purkinje neurons. The expression profile of sonic hedgehog in postnatal stages determines the final size/shape of the cerebellum. Genes affecting the development impact upon the physiological properties of the cerebellar circuits. For instance, receptors are developmentally regulated and their action interferes directly with developmental processes. Another field of research which is expanding relates to very preterm neonates. They are at risk for cerebellar lesions, which may themselves impair the developmental events. Very preterm neonates often show sensori-motor deficits, highlighting another major link between impaired developments and learning deficiencies. Pathways playing a critical role in cerebellar development are likely to become therapeutical targets for several neurodevelopmental disorders. PMID:22291620
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Spinal sensory circuits in motion.
Böhm, Urs Lucas; Wyart, Claire
2016-12-01
The role of sensory feedback in shaping locomotion has been long debated. Recent advances in genetics and behavior analysis revealed the importance of proprioceptive pathways in spinal circuits. The mechanisms underlying peripheral mechanosensation enabled to unravel the networks that feedback to spinal circuits in order to modulate locomotion. Sensory inputs to the vertebrate spinal cord were long thought to originate from the periphery. Recent studies challenge this view: GABAergic sensory neurons located within the spinal cord have been shown to relay mechanical and chemical information from the cerebrospinal fluid to motor circuits. Innovative approaches combining genetics, quantitative analysis of behavior and optogenetics now allow probing the contribution of these sensory feedback pathways to locomotion and recovery following spinal cord injury. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Erfani, Mostafa; Shafiei, Mohammad; Mazidi, Mohammad; Goudarzi, Mostafa
2013-04-01
Somatostatin-derived analogues play an important role in the diagnosis and treatment of neuroendocrine tumors. The aim of this study was to evaluate a new somatostatin analogue designed for labeling with (99m)Tc: [6-hydrazinopyridine-3-carboxylic acid (HYNIC(0)), β-(3-benzothienyl)-Ala (BzThi(3))]-octreotide ([HYNIC]-BOC), using ethylenediamine-N,N'-diacetic acid (EDDA) and tricine as coligands. Synthesis was performed on a solid phase using a standard Fmoc strategy. The HYNIC-peptide conjugate was radiolabeled with (99m)Tc and characterized by ITLC and high-performance liquid chromatography (HPLC). In vitro studies were carried out in sstr2 expressing AR4-2J cell lines. In vivo distribution studies were performed in rats bearing the AR4-2J tumor. The radiolabeled complex could be prepared at high-specific activities and >95% radiochemical yield as determined by HPLC. The peptide conjugate showed high-affinity binding for sstr2. The radioligand showed high and specific internalization into AR4-2J cells (18.19%±0.21% at 4 hours). In vivo distribution studies in rats bearing tumor have shown a receptor-specific uptake of radioactivity in somatostatin receptor-positive organs. After 4 hours, uptake in the AR4-2J tumor was 1.71%±0.36% injected dose per gram tissue (%ID/g). These data show that [(99m)Tc/EDDA/Tricine/HYNIC(0), BzThi(3)]-octreotide is a specific radioligand for the somatostatin receptor-positive tumors and is a suitable candidate for clinical studies.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Litwin, S.D.
Blood mononuclear cells of two individuals having immunodeficiency with thymoma (ID-THY) were cocultured with normal mononuclear cells or treated mononuclear cell fractions in an attempt to correct an imbalance of regulatory cells postulated to be responsible for the failure of pokeweed mitogen-induced Ig synthesis in vitro. Treatment included abrogation of suppressor cell activity by irradiation or incubation with prednisolone in vitro. T cell help was provided by cocultivating lymphocytes of related and unrelated persons, and in some cases autologous treated cells. Ig secretion failed to be induced by any experimental maneuver suggesting that the primary problem in the above ID-THYmore » cells was related to defective or deficient B cells rather than an imbalance of T regulatory cells. Prednisolone treatment in vitro decreased suppressor cell activity in allogeneic cocultures of two ID-THY persons (S1 and S2) but not of an individual (S3) with variable immunodeficiency suggesting heterogeneity of suppressor cells.« less
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Ultrasonic Non-Destructive Testing of Materials.
1980-12-01
k IRIT ’F 1"--A ’I I T . 5 1 ~ Ii I I REPORT DOCUMENTATION PAGE CI- N1Il) I EI. i 0.0 REPORT NUMI3ER i2 GOVT ACCA [. ASINN ,NL t PjAi AFOSR.TR. 80...Peport 107r) thait thi rax!’ t -r lqtionn liron whic-h the- F1 . (.)wan based, was a conseaiencr’ of caal it-"’: -and- ioolniv nd whethe.r suc h ai relation
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Tang, Juan; Chen, Xian; Zhou, Jun; Li, Qunfang; Chen, Guonan; Tang, Dianping
2013-08-07
Multifunctionalized thionine-modified cerium oxide (Thi-CeO2) nanostructures with redox ability and catalytic activity were designed as the bionanolabels for in situ amplified electronic signal of low-abundance protein (carcinoembryonic antigen, CEA, used as a model) based on a cerium oxide-triggered 'one-to-many' catalytic cycling strategy. Initially, the carried CeO2 nanoparticles autocatalytically hydrolyzed the phosphate ester bond of l-ascorbic acid 2-phosphate (AAP) to produce a new reactant (l-ascorbic acid, AA), then the generated AA was electrochemically oxidized by the assembled thionine on the Thi-CeO2, and the resultant product was then reduced back to AA by the added tris(2-carboxyethy)phosphine (TCEP). The catalytic cycling could be re-triggered by the thionine and TCEP, resulting in amplification of the electrochemical signal. Under the optimized conditions, the electrochemical immunosensor exhibited a wide linear range of 0.1 pg mL(-1) to 80 ng mL(-1) with a low detection limit of 0.08 pg mL(-1) CEA at the 3σblank level. In addition, the methodology was evaluated for the analysis of clinical serum samples, and was in good accordance with values obtained using the commercialized enzyme-linked immunosorbent assay (ELISA) method.
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1981-12-01
Steven W. Congdon Physical Science John Collis Geography Abram J. Nicholson Civil Engineering Franklin L. Snitz Environmental Chemistry 6.02 The data...Isllfeebe tP deil,1t1v -, i .1d IO~ 1 , ’, , o- , ,,r-nS ,,n J. Thi pteId h,11 ’- 1 tr ts.-d o lrrf¢t . withot 5. wrlfan aore’sav of the land Resource
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Cantisani, Vito; Maceroni, Piero; D'Andrea, Vito; Patrizi, Gregorio; Di Segni, Mattia; De Vito, Corrado; Grazhdani, Hektor; Isidori, Andrea M; Giannetta, Elisa; Redler, Adriano; Frattaroli, Fabrizio; Giacomelli, Laura; Di Rocco, Giorgio; Catalano, Carlo; D'Ambrosio, Ferdinando
2016-05-01
To assess whether ultrasound elastography (USE) with strain ratio increases diagnostic accuracy of Doppler ultrasound in further characterisation of cytologically Thy3 thyroid nodules. In two different university diagnostic centres, 315 patients with indeterminate cytology (Thy3) in thyroid nodules aspirates were prospectively evaluated with Doppler ultrasound and strain ratio USE before surgery. Ultrasonographic features were analysed separately and together as ultrasound score, to assess sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Receiver operating characteristic (ROC) curves to identify optimal cut-off value of the strain ratio were also provided. Diagnosis on a surgical specimen was considered the standard of reference. Higher strain ratio values were found in malignant nodules, with an optimum strain ratio cut-off of 2.09 at ROC analysis. USE with strain ratio showed 90.6% sensitivity, 93% specificity, 82.8% PPV, 96.4% NPV, while US score yielded a sensitivity of 52.9%, specificity of 84.3%, PPV 55.6% and NPV 82.9%. The diagnostic gain with strain ratio was statistically significant as proved by ROC areas, which was 0.9182 for strain ratio and 0.6864 for US score. USE with strain ratio should be considered a useful additional tool to colour-Doppler US, since it improves characterisation of thyroid nodules with indeterminate cytology. • Strain ratio measurements improve differentiation of thyroid nodules with indeterminate cytology • Elastography with strain ratio is more reliable than ultrasound features and ultrasound score • Strain ratio may help to better select patients with Thy 3 nodules candidate for surgery.
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Jeon, Sun Woo; Kim, Kyu Seok; Nam, Hae Jeong
2012-07-01
The aim of this study was to investigate the long-term effect of acupuncture for treatment of tinnitus. This study is a randomized, patient- and assessor-blind, sham-acupuncture-controlled, pilot trial. Participants were adults (18-60 years old) with chronic (≥6 months), unilateral tinnitus, and without moderate or severe hearing loss. Thirty-three (33) participants were randomized to one of two treatment groups: real acupuncture and sham-acupuncture (nonmeridian; no specific response, de qi). Participants received 10 sessions of acupuncture treatment (twice a week for 5 weeks), and usual patient care education. The subjective outcome was the score of Tinnitus Handicap Inventory (THI) and Visual Analogue Scale (VAS) from baseline to 3 months after. Pure Tone Average (PTA) and Speech Discrimination (SD) from baseline to 3 months after were assessed as objective outcomes. A significant interaction between time and group in VAS (p=0.017) was evident, but not in THI, PTA, and SD scores. THI showed significant improvement at the end of treatment and 3 months after, compared to baseline, in real acupuncture (p=0.004). In SD, a significant long-term effect of real acupuncture was observed until 3 months after (p=0.011). However, the effect of real acupuncture in PTA was not maintained until 3 months after the end of treatment. No significant difference in the sham-acupuncture treatment group was evident. No statistical difference in any outcome was observed between real and sham acupuncture. Only in the mean percent change of VAS, real acupuncture showed statistical significance, compared with sham-acupuncture from baseline to 3 months after (p=0.019). Through evaluation of subjective (THI and VAS) and objective outcomes (PTA and SD), this study demonstrates the long-term effect of real acupuncture.
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One-year follow up of patients with chronic tinnitus treated with left temporoparietal rTMS.
Khedr, E M; Rothwell, J C; El-Atar, A
2009-03-01
Although there are a number of positive reports on the therapeutic effects of repetitive transcranial magnetic stimulation (rTMS) for treatment of tinnitus, there are few details about the duration of treatment effects or the relative efficiency of different rTMS protocols. Sixty six patients with chronic tinnitus were divided into four groups, receiving sham rTMS, 1, 10 and 25 Hz rTMS applied each day for 10 days over left temporoparietal cortex. They were followed up at 4 months and 1 year using the tinnitus questionnaire [Tinnitus Handicap Inventory(THI)] and self ratings of annoyance as well as measures of residual inhibition. A two factor anova revealed a significant 'rTMS' x 'time' interaction indicating that real and sham rTMS had different effects on the THI scale and annoyance of tinnitus (P = 0.026 and 0.046 respectively). After 1 year, the tinnitus was absent in one or both ears of 10 patients who had received real rTMS: one of these was in the 1 Hz group, four patients were in the 10 Hz group and five patients were in the 25 Hz group. Some patients show a lasting benefit at 1 year after 10 days of rTMS treatment. It appears that treatment at 10 or 25 Hz may be more beneficial than at 1 Hz, although more work is necessary to validate this conclusion.
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Shehab-El-Deen, Mohamed Ahmed M M; Fadel, Moustafa S; Van Soom, Ann; Saleh, Sherif Y; Maes, Dominiek; Leroy, Jo L M R
2010-08-01
The current study aimed to investigate the circadian rhythm of blood metabolic parameters associated with summer heat stress (HS) in dairy cows. Ten healthy lactating Holstein Friesian cows were followed during HS for three successive days at six different time points. Blood was sampled from each cow starting from 07:00 AM: ; at 4-h intervals. Ambient air temperature and relative humidity were recorded, and temperature-humidity index (THI) was calculated as well. Respiration rate (RR) and rectal temperature (RT) were recorded for each cow at the time of blood sampling. Concentrations of glucose, non-esterified fatty acids (NEFA), total cholesterol (TC) and urea were measured in each blood sample. The THI values were >68 at all times of the day, and the highest values were recorded at 11:00 AM: , 03:00 PM: and 07:00 PM: (80.9, 83.7, and 80.8, respectively). All the cows showed a significantly higher RR and RT coinciding with higher THI values (93 +/- 4 and 39.6 +/- 0.1; 90.2 +/- 3.4, and 40.1 +/- 0.1; 87.6 +/- 4.1, and 39.8 +/- 0.1, respectively, P < 0.05). The concentrations of glucose were the lowest at 11:00 AM: and 03:00 PM: (3.75 +/- 0.1 and 3.44 +/- 0.1 mmol/L, respectively, P < 0.05). Decreased glucose concentrations coincided with increased NEFA concentrations, (0.43 +/- 0.01 and 0.56 +/- 0.02 mmol/L, respectively, P < 0.05), and were highly negatively correlated (r = -0.50, P < 0.001). The highest urea and TC concentrations were registered at 11:00 AM: (6.11 +/- 0.15 mmol/L and 109.9 +/- 2.2 mg/dl, respectively) whereas the lowest urea and TC values were recorded at 03:00 AM: (4.97 +/- 0.18 mmol/L and 99.5 +/- 1.7 mg/dl, respectively, P < 0.05). The results of the present study indicate that there was a circadian variation in glucose, NEFA, urea, and TC resulting in the most unfavorable metabolic condition during the hottest moment of the day in dairy cattle. Earlier work revealed that HS-metabolic changes are reflected in the follicular fluid. The circadian changes observed in the present study associated with HS may imply that also the microenvironment of the oocyte is affected.
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Molecular Analysis of Sensory Axon Branching Unraveled a cGMP-Dependent Signaling Cascade.
Dumoulin, Alexandre; Ter-Avetisyan, Gohar; Schmidt, Hannes; Rathjen, Fritz G
2018-04-24
Axonal branching is a key process in the establishment of circuit connectivity within the nervous system. Molecular-genetic studies have shown that a specific form of axonal branching—the bifurcation of sensory neurons at the transition zone between the peripheral and the central nervous system—is regulated by a cyclic guanosine monophosphate (cGMP)-dependent signaling cascade which is composed of C-type natriuretic peptide (CNP), the receptor guanylyl cyclase Npr2, and cGMP-dependent protein kinase Iα (cGKIα). In the absence of any one of these components, neurons in dorsal root ganglia (DRG) and cranial sensory ganglia no longer bifurcate, and instead turn in either an ascending or a descending direction. In contrast, collateral axonal branch formation which represents a second type of axonal branch formation is not affected by inactivation of CNP, Npr2, or cGKI. Whereas axon bifurcation was lost in mouse mutants deficient for components of CNP-induced cGMP formation; the absence of the cGMP-degrading enzyme phosphodiesterase 2A had no effect on axon bifurcation. Adult mice that lack sensory axon bifurcation due to the conditional inactivation of Npr2-mediated cGMP signaling in DRG neurons demonstrated an altered shape of sensory axon terminal fields in the spinal cord, indicating that elaborate compensatory mechanisms reorganize neuronal circuits in the absence of bifurcation. On a functional level, these mice showed impaired heat sensation and nociception induced by chemical irritants, whereas responses to cold sensation, mechanical stimulation, and motor coordination are normal. These data point to a critical role of axon bifurcation for the processing of acute pain perception.
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Rotigotine protects against glutamate toxicity in primary dopaminergic cell culture.
Oster, Sandra; Radad, Khaled; Scheller, Dieter; Hesse, Marlen; Balanzew, Wladimir; Reichmann, Heinz; Gille, Gabriele
2014-02-05
In Parkinson disease the degeneration of dopaminergic neurones is believed to lead to a disinhibition of the subthalamic nucleus thus increasing the firing rate of the glutamatergic excitatory projections to the substantia nigra. In consequence, excessive glutamatergic activity will cause excitotoxicity and oxidative stress. In the present study we investigated mechanisms of glutamate toxicity and the neuroprotective potential of the dopamine agonist rotigotine towards dopaminergic neurones in mouse mesencephalic primary culture. Glutamate toxicity was mediated by the N-methyl-d-aspartic acid (NMDA) receptor and accompanied by a strong calcium influx into dopaminergic neurones for which the L-type voltage-sensitive calcium channels play an important role. The rate of superoxide production in the culture was highly increased. Deleterious nitric oxide production did not participate in glutamate-mediated excitotoxicity. Pretreatment of cultures with rotigotine significantly increased the survival of dopaminergic neurones exposed to glutamate. Rotigotine exerted its protective effects via dopamine receptor stimulation (presumably via dopamine D3 receptor) and decreased significantly the production of superoxide radicals. When cultures were preincubated with Phosphoinositol 3-Kinase (PI3K) inhibitors the protective effect of rotigotine was abolished suggesting a decisive role of the PI3K/Akt pathway in rotigotine-mediated neuroprotection. Consistently, exposure to rotigotine induced the activation of Akt by phosphorylation followed by phosphorylation, and thus inactivation, of the pro-apoptotic factor glycogen synthase kinase-3-beta (GSK-3-β). Taken together, our work contributed to elucidating the mechanisms of glutamate toxicity in mesencephalic culture and unravelled the signalling pathways associated with rotigotine-induced neuroprotection against glutamate toxicity in primary dopaminergic cultures. Copyright © 2013 Elsevier B.V. All rights reserved.
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Meyer, Néva P; Carrillo-Baltodano, Allan; Moore, Richard E; Seaver, Elaine C
2015-01-01
Reconstructing the evolutionary history of nervous systems requires an understanding of their architecture and development across diverse taxa. The spiralians encompass diverse body plans and organ systems, and within the spiralians, annelids exhibit a variety of morphologies, life histories, feeding modes and associated nervous systems, making them an ideal group for studying evolution of nervous systems. We describe nervous system development in the annelid Capitella teleta (Blake JA, Grassle JP, Eckelbarger KJ. Capitella teleta, a new species designation for the opportunistic and experimental Capitella sp. I, with a review of the literature for confirmed records. Zoosymposia. 2009;2:25-53) using whole-mount in situ hybridization for a synaptotagmin 1 homolog, nuclear stains, and cross-reactive antibodies against acetylated α-tubulin, 5-HT and FMRFamide. Capitella teleta is member of the Sedentaria (Struck TH, Paul C, Hill N, Hartmann S, Hosel C, Kube M, et al. Phylogenomic analyses unravel annelid evolution. Nature. 2011;471:95-8) and has an indirectly-developing, lecithotrophic larva. The nervous system of C. teleta shares many features with other annelids, including a brain and a ladder-like ventral nerve cord with five connectives, reiterated commissures, and pairs of peripheral nerves. Development of the nervous system begins with the first neurons differentiating in the brain, and follows a temporal order from central to peripheral and from anterior to posterior. Similar to other annelids, neurons with serotonin-like-immunoreactivity (5HT-LIR) and FMRFamide-like-immunoreactivity (FMRF-LIR) are found throughout the brain and ventral nerve cord. A small number of larval-specific neurons and neurites are present, but are visible only after the central nervous system begins to form. These larval neurons are not visible after metamorphosis while the rest of the nervous system is largely unchanged in juveniles. Most of the nervous system that forms during larvogenesis in C. teleta persists into the juvenile stage. The first neurons differentiate in the brain, which contrasts with the early formation of peripheral, larval-specific neurons found in some spiralian taxa with planktotrophic larvae. Our study provides a clear indication that certain shared features among annelids - e.g., five connectives in the ventral nerve cord - are only visible during larval stages in particular species, emphasizing the need to include developmental data in ancestral character state reconstructions. The data provided in this paper will serve as an important comparative reference for understanding evolution of nervous systems, and as a framework for future molecular studies of development.
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Huang, Ying; Zitta, Karina; Bein, Berthold; Steinfath, Markus; Albrecht, Martin
2013-01-01
SUMMARY Ischemia-reperfusion injury and tissue hypoxia are of high clinical relevance because they are associated with various pathophysiological conditions such as myocardial infarction and stroke. Nevertheless, the underlying mechanisms causing cell damage are still not fully understood, which is at least partially due to the lack of cell culture systems for the induction of rapid and transient hypoxic conditions. The aim of the study was to establish a model that is suitable for the investigation of cellular and molecular effects associated with transient and long-term hypoxia and to gain insights into hypoxia-mediated mechanisms employing a neuronal culture system. A semipermeable membrane insert system in combination with the hypoxia-inducing enzymes glucose oxidase and catalase was employed to rapidly and reversibly generate hypoxic conditions in the culture medium. Hydrogen peroxide assays, glucose measurements and western blotting were performed to validate the system and to evaluate the effects of the generated hypoxia on neuronal IMR-32 cells. Using the insert-based two-enzyme model, hypoxic conditions were rapidly induced in the culture medium. Glucose concentrations gradually decreased, whereas levels of hydrogen peroxide were not altered. Moreover, a rapid and reversible (onoff) generation of hypoxia could be performed by the addition and subsequent removal of the enzyme-containing inserts. Employing neuronal IMR-32 cells, we showed that 3 hours of hypoxia led to morphological signs of cellular damage and significantly increased levels of lactate dehydrogenase (a biochemical marker of cell damage). Hypoxic conditions also increased the amounts of cellular procaspase-3 and catalase as well as phosphorylation of the pro-survival kinase Akt, but not Erk1/2 or STAT5. In summary, we present a novel framework for investigating hypoxia-mediated mechanisms at the cellular level. We claim that the model, the first of its kind, enables researchers to rapidly and reversibly induce hypoxic conditions in vitro without unwanted interference of the hypoxia-inducing agent on the cultured cells. The system could help to further unravel hypoxia-associated mechanisms that are clinically relevant in various tissues and organs. PMID:24046359
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ERIC Educational Resources Information Center
Middei, Silvia; Roberto, Anna; Berretta, Nicola; Panico, Maria Beatrice; Lista, Simone; Bernardi, Giorgio; Mercuri, Nicola B.; Ammassari-Teule, Martine; Nistico, Robert
2010-01-01
B6-Tg/Thy1APP23Sdz (APP23) mutant mice exhibit neurohistological hallmarks of Alzheimer's disease but show intact basal hippocampal neurotransmission and synaptic plasticity. Here, we examine whether spatial learning differently modifies the structural and electrophysiological properties of hippocampal synapses in APP23 and wild-type mice. While…
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Domination Problem for Vector Measures and Applications to Nonstationary Processes.
1981-09-23
meas- ures, which have p-semi-variation finite, I a p a 2 . The work here do- pieda in part on an inequality of Grothendieck- Pietsch . The rest of the...7], Corol. 2 to Thi. 4.3 and Prop. 3.1, the Latter is the Grothendieck- Pietsch inequality alluded to in the Introduction), the space I - C(S) being
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Air and Space Power Journal. Volume 26, Number 1, January-February 2012
2012-01-01
Deployment Tasking Process Doesn’t Do . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 Lt... processes that can fundamentally alter the way we do business. We cannot implement this essential shift merely by trimming “around the edges” and making...Journal. This wor- thy initiative highlights the process of turning challenges into opportuni- ties. ASPJ is charged with providing a forum in which
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U.S. EPA, Pesticide Product Label, VISCO 1153, 06/16/1989
2011-04-19
... 0, •• ""i., IS p,"hi.it ••. PE~TICIDE OISPOSAl: P,stiei', Kast,s Irt t.XIC I",,,, is,.s.1 .f Ixe.ss ~.stici' •• S,'I, "ix t." •• , rits.t. is •• i.lltl ••• f F ••• ,.I L •• I' thiS. ...
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Novel method for in vitro depletion of T cells by monoclonal antibody-targeted photosensitization.
Berki, T; Németh, P
1998-02-01
An immunotargeting method (called photo-immunotargeting) has been developed for selective in vitro cell destruction. The procedure combines the photosensitizing (toxic) effect of light-induced dye-molecules, e.g., hematoporphyrin (HP) and the selective binding ability of monoclonal antibodies (mAb) to cell surface molecules. The photosensitizer HP molecules were covalently attached to monoclonal antibodies (a-Thy-1) recognizing an antigen on the surface of T lymphocytes, and used for T cell destruction. To increase the selectivity of the conventional targeting methods, a physical activation step (local light irradiation) as a second degree of specificity was employed. The HP in conjugated form was sufficient to induce T cell (thymocytes, EL-4 cell line) death after irradiation at 400 nm, at tenfold lower concentration compared to the photosensitizing effect of unbound HP. The selective killing of T lymphocytes (bearing the Thy-1 antigen) in a mixed cell population was demonstrated after a treatment with the phototoxic conjugate and light irradiation. This method can be useful for selective destruction of one population (target cell) in an in vitro heterogeneous cell mixture, e.g., in bone marrow transplants for T cell depletion to avoid graft vs. host reaction.
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Chronic Ethanol During Adolescence Impacts Corticolimbic Dendritic Spines and Behavior.
Jury, Nicholas J; Pollack, Gabrielle A; Ward, Meredith J; Bezek, Jessica L; Ng, Alexandra J; Pinard, Courtney R; Bergstrom, Hadley C; Holmes, Andrew
2017-07-01
Risk for alcohol use disorders (AUDs) in adulthood is linked to alcohol drinking during adolescence, but understanding of the neural and behavioral consequences of alcohol exposure during adolescence remains incomplete. Here, we examined the neurobehavioral impact of adolescent chronic intermittent EtOH (CIE) vapor exposure in mice. C57BL/6J-background Thy1-EGFP mice were CIE-exposed during adolescence or adulthood and examined, as adults, for alterations in the density and morphology of dendritic spines in infralimbic (IL) cortex, prelimbic (PL) cortex, and basolateral amygdala (BLA). In parallel, adolescent- and adult-exposed C57BL/6J mice were tested as adults for 2-bottle EtOH drinking, sensitivity to EtOH intoxication (loss of righting reflex [LORR]), blood EtOH clearance, and measures of operant responding for food reward. CIE during adolescence decreased IL neuronal spine density and increased the head width of relatively wide-head IL and BLA spines, whereas CIE decreased head width of relatively narrow-head BLA spines. Adolescents had higher EtOH consumption prior to CIE than adults, while CIE during adulthood, but not adolescence, increased EtOH consumption relative to pre-CIE baseline. CIE produced a tolerance-like decrease in LORR sensitivity to EtOH challenge, irrespective of the age at which mice received CIE exposure. Mice exposed to CIE during adolescence, but not adulthood, required more sessions than AIR controls to reliably respond for food reward on a fixed-ratio (FR) 1, but not subsequent FR3, reinforcement schedule. On a progressive ratio reinforcement schedule, break point responding was higher in the adolescent- than the adult-exposed mice, regardless of CIE. Finally, footshock punishment markedly suppressed responding for reward in all groups. Exposure to CIE during adolescence altered dendritic spine density and morphology in IL and BLA neurons, in parallel with a limited set of behavioral alterations. Together, these data add to growing evidence that key corticolimbic circuits are vulnerable to the effects of alcohol during adolescence, with lasting, potentially detrimental, consequences for behavior. Copyright © 2017 by the Research Society on Alcoholism.
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Chronic ethanol during adolescence impacts corticolimbic dendritic spines and behavior
Jury, Nicholas J.; Pollack, Gabrielle A.; Ward, Meredith J.; Bezek, Jessica L.; Ng, Alexandra J.; Pinard, Courtney R.; Bergstrom, Hadley C.; Holmes, Andrew
2017-01-01
Background Risk for alcohol use disorders (AUDs) is linked to alcohol drinking during adolescence, but understanding of the neural and behavioral consequences of alcohol exposure during adolescence remains incomplete. Here, we examined the neurobehavioral impact of adolescent chronic intermittent EtOH (CIE) vapor exposure in mice. Methods C57BL/6J-background Thy1-EGFP mice were CIE-exposed during adolescence or adulthood and examined, as adults, for alterations in the density and morphology of dendritic spines in infralimbic cortex (IL), prelimbic cortex (PL) and basolateral amygdala (BLA). In parallel, adolescent- and adult-exposed C57BL/6J mice were tested as adults for two-bottle EtOH drinking, sensitivity to EtOH intoxication (loss of righting reflex, LORR), blood EtOH clearance, and measures of operant responding for food reward. Results CIE during adolescence decreased IL neuronal spine density and increased the head-width of relatively wide-head IL and BLA spines, whereas CIE decreased head-width of relatively narrow-head BLA spines. Adolescents had higher EtOH consumption prior to CIE than adults, while CIE during adulthood, but not adolescence, increased EtOH consumption relative to pre-CIE baseline. CIE produced a tolerance-like decrease in LORR sensitivity to EtOH challenge, irrespective of the age at which mice received CIE exposure. Mice exposed to CIE during adolescence, but not adulthood, required more sessions than AIR controls to reliably respond for food reward on a fixed-ratio (FR1), but not subsequent FR3, reinforcement schedule. On a progressive ratio reinforcement schedule, breakpoint responding was higher in the adolescent- than the adult-exposed mice, regardless of CIE. Finally, footshock-punishment markedly suppressed responding for reward in all groups. Conclusions Exposure to CIE during adolescence altered dendritic spine density and morphology in IL and BLA neurons, in parallel with a limited set of behavioral alterations. Together, these data add to growing evidence that key corticolimbic circuits are vulnerable to the effects of alcohol during adolescence, with lasting, potentially detrimental, consequences for behavior. PMID:28614590
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Buckle, A. M.; Mottram, R.; Pierce, A.; Lucas, G. S.; Russell, N.; Miyan, J. A.; Whetton, A. D.
2000-01-01
BACKGROUND: Chronic Myeloid Leukaemia (CML) is characterised by the chromosomal translocation resulting in expression of the Bcr-Abl protein tyrosine kinase (PTK) in early stem cells and their progeny. However the precise nature of Bcr-Abl effects in primitive CML stem cells remains a matter of active debate. MATERIALS AND METHODS: Extremely primitive Bcr-Abl fusion positive cells were purified from patients with CML using multiparameter flow cytometric analysis of CD34, Thy, and lineage marker (Lin) expression, plus rhodamine-123 (Rh-123) brightness. Progenitor cells of increasing maturity were examined for cycling status by flow cytometry and their proliferative status directly correlated with cell phenotype. The activation status of a key transcription factor, signal transducers and activators of transcription (STAT-5), was also analyzed by immunocytochemistry. RESULTS: The most primitive stem cells currently defined (CD34+Lin-Thy+ Rh-1231o) were present as a lower proportion of the stem cell compartment (CD34+Lin-) of CML patients at presentation than of normal individuals (2.3% +/- 0.4 compared with 5.1% +/- 0.6 respectively). Conversely there was a significantly higher proportion of the more mature cells (CD34+Lin-Thy-Rh-123 hi) in CML patients than in normal individuals (79.3 +/- 1.8 compared with 70.9 +/- 3.3). No primitive subpopulation of CML CD34+Lin- cells was cycling to a significantly greater degree than cells from normal donors, in fact, late progenitor cells (CD34+Lin+) were cycling significantly less in CML samples than normal samples. STAT5, however, was observed to be activated in CML cells. CONCLUSIONS: We conclude that no subpopulation of CML stem cells displays significantly increased cell cycling. Thus, increased cycling cannot be a direct consequence of Bcr-Abl PTK acquisition in highly enriched stem cells from patients with CML. In vivo CML need not be considered a disease of unbridled stem cell proliferation, but a subtle defect in the balance between self renewal and maturation. PMID:11126203
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1979-09-01
CAN E GALLEO 3y: GCOSF G(,jPSF GCUNPFk’ E NT py_ SC.A5S4 A 1’ _ _S FG MEN T ( O OESC I £NITE, TE. - UNPACK SUBFILE OR OAE LIST LENr ,TH1 75...SEGMENTS (PR2) DESCS SET LINE MODE IN DRAFTING LENr ,THI ------- 5 --lANGUAGES FTN . ...... - HAS NO SECONDARY ENTRY POINTS CAN BE CALLED BYI PRTADLE B DSTR
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Checking for Circular Dependencies in Distributed Stream Programs
2011-08-29
extensions to express new complexities more conve- nient. Teleport messaging ( TMG ) in the StreamIt language [30] is an example. 1.1 StreamIt Language...dynamicities to an FIR computation Thies et al. in [30] give a TMG model for distributed stream pro- grams. TMG is a mechanism that implements control...messages for stream graphs. The TMG mechanism is designed not to interfere with original dataflow graphs’ structures and scheduling, therefore a key
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The influence of psychological factors on tinnitus severity.
Milerová, Jana; Anders, Martin; Dvořák, Tomáš; Sand, Philipp G; Königer, Stefanie; Langguth, Berthold
2013-01-01
Subjective tinnitus is a frequent symptom characterized by perception of sound in the absence of a corresponding external stimulus. Although many people learn to live with tinnitus, some find it severely debilitating. Why tinnitus is debilitating in some patients, but not in others, is still incompletely understood. We aimed to assess the influence of different aspects of psychological distress on perceived tinnitus severity. Three hundred seventeen patients diagnosed with chronic subjective tinnitus at two university clinics completed the Tinnitus Handicap Inventory (THI), the Tinnitus Questionnaire (TQ) and the Symptom Check List-90-Revised. The influence of the different dimensions of psychological distress on perceived tinnitus severity was statistically evaluated. Both THI and TQ scores were significantly influenced by gender, site and the dimension "depression". In addition, TQ scores were significantly influenced by age and "somatization," whereas "hostility" had an impact on THI scores only. Psychological aspects as well as sociodemographic variables had a significant influence on both TQ scores. However, our results indicate, that these scales reflect emotional distress of tinnitus sufferers differently. This should be taken into consideration in the use of these scales as screening tools for assessment of tinnitus handicap. Copyright © 2013. Published by Elsevier Inc.
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Socioeconomic status of parents with children participating on youth club sport teams.
Post, Eric G; Green, Nicole E; Schaefer, Daniel A; Trigsted, Stephanie M; Brooks, M Alison; McGuine, Timothy A; Watson, Andrew M; Bell, David R
2018-05-17
To describe the socioeconomic status, measured by household income and educational attainment, of parents with children participating on youth club sport teams. Cross-sectional survey. Local sport events. 949 parents (571 female) of youth athletes between 10 and 18 years old were recruited at club team events and practices to complete an anonymous questionnaire. SES variables included total household income (THI) and educational attainment. Sport specialization was classified as low, moderate, or high using a previously utilized 3-point specialization scale. Chi-square tests were used to compare frequencies SES categories with child specialization. Parents reported spending a median of 1500 [500-3000] USD per year on their children's club sports activities. Most parents reported a THI greater than 100,000 USD per year and a bachelor's degree or higher level of education. Parents in the higher THI categories were more likely to have a child that is highly specialized in one sport. The current youth sports system in the United States, which emphasizes year-round participation on club teams, may be limiting the participation of families without the resources to participate in this system. Copyright © 2018 Elsevier Ltd. All rights reserved.
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Yilmaz, Ömer H.; Kiel, Mark J.; Morrison, Sean J.
2006-01-01
Recent advances have increased the purity of hematopoietic stem cells (HSCs) isolated from young mouse bone marrow. However, little attention has been paid to the purity of HSCs from other contexts. Although Thy-1lowSca-1+Lineage-c-kit+ cells from young bone marrow are highly enriched for HSCs (1 in 5 cells gives long-term multilineage reconstitution after transplantation into irradiated mice), the same population from old, reconstituted, or cytokine-mobilized mice engrafts much less efficiently (1 in 78 to 1 in 185 cells gives long-term multilineage reconstitution). To test whether we could increase the purity of HSCs isolated from these contexts, we examined the SLAM family markers CD150 and CD48. All detectable HSCs from old, reconstituted, and cyclophosphamide/G-CSF-mobilized mice were CD150+CD48-, just as in normal young bone marrow. Thy-1lowSca-1+Lineage-c-kit+ cells from old, reconstituted, or mobilized mice included mainly CD48+ and/or CD150- cells that lacked reconstituting ability. CD150+CD48-Sca-1+Lineage-c-kit+ cells from old, reconstituted, or mobilized mice were much more highly enriched for HSCs, with 1 in 3 to 1 in 7 cells giving long-term multilineage reconstitution. SLAM family receptor expression is conserved among HSCs from diverse contexts, and HSCs from old, reconstituted, and mobilized mice engraft relatively efficiently after transplantation when contaminating cells are eliminated. PMID:16219798
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Yilmaz, Omer H; Kiel, Mark J; Morrison, Sean J
2006-02-01
Recent advances have increased the purity of hematopoietic stem cells (HSCs) isolated from young mouse bone marrow. However, little attention has been paid to the purity of HSCs from other contexts. Although Thy-1 low Sca-1+ Lineage- c-kit+ cells from young bone marrow are highly enriched for HSCs (1 in 5 cells gives long-term multilineage reconstitution after transplantation into irradiated mice), the same population from old, reconstituted, or cytokine-mobilized mice engrafts much less efficiently (1 in 78 to 1 in 185 cells gives long-term multilineage reconstitution). To test whether we could increase the purity of HSCs isolated from these contexts, we examined the SLAM family markers CD150 and CD48. All detectable HSCs from old, reconstituted, and cyclophosphamide/G-CSF-mobilized mice were CD150+ CD48-, just as in normal young bone marrow. Thy-1 low Sca-1+ Lineage- c-kit+ cells from old, reconstituted, or mobilized mice included mainly CD48+ and/or CD150- cells that lacked reconstituting ability. CD150+ CD48- Sca-1+ Lineage- c-kit+ cells from old, reconstituted, or mobilized mice were much more highly enriched for HSCs, with 1 in 3 to 1 in 7 cells giving long-term multilineage reconstitution. SLAM family receptor expression is conserved among HSCs from diverse contexts, and HSCs from old, reconstituted, and mobilized mice engraft relatively efficiently after transplantation when contaminating cells are eliminated.
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Mattachini, Gabriele; Bava, Luciana; Sandrucci, Anna; Tamburini, Alberto; Riva, Elisabetta; Provolo, Giorgio
2017-08-01
This study aimed to examine the influence of feed delivery frequency and environmental conditions on daily time budget of lactating dairy cows. The study was carried out in two commercial dairy farms with Holstein herds. Fifty lactating dairy cows milked in automatic milking units (AMS farm) and 96 primiparous lactating dairy cows milked in a conventional milking parlour (conventional farm) were exposed to different frequencies of feed delivery replicated in different periods of the year (warm and mild) that were characterized by different temperature-humidity indices (THI). On each farm, feeding treatments consisted of two different feed delivery frequencies (1× and 2× on the AMS farm; 2× and 3× on the conventional farm). All behaviours of the cows were monitored for the last 8 d of each treatment period using continuous video recording. The two data sets from different farm systems were considered separately for analysis. On both farms, environmental conditions expressed as THI affected time budgets and the pattern of the behavioural indices throughout the day. The variation in the frequency of feed delivery seems to affect the cow's time budget only in a limited way. Standing time of cows on the conventional farm and the time spent by cows in the milking waiting area on the AMS farm both increased in response to increased feeding frequency. Although feed delivery frequency showed limited influence on cow's time budget, the effect on standing time could be carefully considered, especially on farms equipped with AMS where the type of cow traffic system (e.g., milking first) might amplify the negative consequences of more frequent feed delivery. Further investigations are required to evaluate the effect of THI and feed delivery frequency on other aspects of behavioural activity.
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Comparative Toxicity of Preservatives on Immortalized Corneal and Conjunctival Epithelial Cells
Ahdoot, Michael; Marcus, Edward; Asbell, Penny A.
2009-01-01
Abstract Purpose Nearly all eye drops contain preservatives to decrease contamination. Nonpreservatives such as disodium-ethylene diamine tetra-acetate (EDTA) and phosphate-buffered saline are also regularly added as buffering agents. These components can add to the toxicity of eye drops and cause ocular surface disease. To evaluate the potential toxicity of these common components and their comparative effects on the ocular surface, a tissue culture model utilizing immortalized corneal and conjunctival epithelial cells was utilized. Methods Immortalized human conjunctival and corneal epithelial cells were grown. At confluency, medium was replaced with 100 μL of varying concentrations of preservatives: benzalkonium chloride (BAK), methyl paraben (MP), sodium perborate (SP), chlorobutanol (Cbl), and stabilized thimerosal (Thi); varying concentrations of buffer: EDTA; media (viable control); and formalin (dead control). After 1 h, solutions were replaced with 150 μL of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazonium bromide). After 4 h, solutions decanted, 100 μL of acid isopropanol added, and the optical density determined at 572 nm to evaluate cell viability. Results Conjunctival and corneal cell toxicity was seen with all preservatives. Depending upon concentration, BAK exhibited from 56% to 89% toxicity. In comparison, Cbl exhibited from 50% to 86%, MP from 30% to 76%, SP from 23% to 59%, and Thi from 70% to 95%. EDTA with minimal toxicity (from 6% to 59%) was indistinguishable from SP. Conclusions Generally, the order of decreasing toxicity at the most commonly used concentrations: Thi (0.0025%) > BAK (0.025%) > Cbl (0.25%) > MP (0.01%) > SP (0.0025%) ≈ EDTA (0.01%). Even at low concentration, these agents will cause some degree of ocular tissue damage. PMID:19284328
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Nidumolu, Uday; Crimp, Steven; Gobbett, David; Laing, Alison; Howden, Mark; Little, Stephen
2014-08-01
The Murray dairy region produces approximately 1.85 billion litres of milk each year, representing about 20 % of Australia's total annual milk production. An ongoing production challenge in this region is the management of the impacts of heat stress during spring and summer. An increase in the frequency and severity of extreme temperature events due to climate change may result in additional heat stress and production losses. This paper assesses the changing nature of heat stress now, and into the future, using historical data and climate change projections for the region using the temperature humidity index (THI). Projected temperature and relative humidity changes from two global climate models (GCMs), CSIRO MK3.5 and CCR-MIROC-H, have been used to calculate THI values for 2025 and 2050, and summarized as mean occurrence of, and mean length of consecutive high heat stress periods. The future climate scenarios explored show that by 2025 an additional 12-15 days (compared to 1971 to 2000 baseline data) of moderate to severe heat stress are likely across much of the study region. By 2050, larger increases in severity and occurrence of heat stress are likely (i.e. an additional 31-42 moderate to severe heat stress days compared with baseline data). This increasing trend will have a negative impact on milk production among dairy cattle in the region. The results from this study provide useful insights on the trends in THI in the region. Dairy farmers and the dairy industry could use these results to devise and prioritise adaptation options to deal with projected increases in heat stress frequency and severity.
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Moderate summer heat stress does not modify immunological parameters of Holstein dairy cows
NASA Astrophysics Data System (ADS)
Lacetera, Nicola; Bernabucci, Umberto; Ronchi, Bruno; Scalia, Daniela; Nardone, Alessandro
2002-02-01
The study was undertaken during spring and summer months in a territory representative of the Mediterranean climate to assess the effects of season on some immunological parameters of dairy cows. Twenty Holstein cows were used. Eleven of those cows gave birth during spring; the remaining nine cows gave birth in summer. The two groups of cows were homogeneous for parity. Values of air temperatures and relative humidity were recorded both during spring and summer, and were utilized to calculate the temperature humidity index (THI). One week before the expected calving, rectal temperatures and respiratory rates of the cows were recorded (1500 hours), and cell-mediated immunity was assessed by measuring the proliferation of mitogen-stimulated peripheral blood mononuclear cells (PBMC). Within 3 h of calving, one colostrum sample was taken from each cow and analysed to determine content of immunoglobulin (Ig) G1, IgG2, IgM and IgA. At 48 h after birth, passive immunization of the calves was assessed by measuring total serum IgG. During summer, daytime (0900-2000 hours) THI values were above the upper critical value of 72 [75.2, (SD 2.6)] indicating conditions that could represent moderate heat stress. That THI values were able to predict heat stress was confirmed by the values of rectal temperatures and respiratory rates, which were higher ( P < 0.05 and P < 0.001 respectively) during summer. Proliferation of PBMC, the colostral concentration of Ig fractions and serum levels of IgG in their respective offspring did not differ between spring and summer cows. Results indicated that moderate heat stress due to the hot Mediterranean summer does not modify cell-mediated immunity, the protective value of colostrum and passive immunization of the offspring in dairy cows.
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NASA Astrophysics Data System (ADS)
Nidumolu, Uday; Crimp, Steven; Gobbett, David; Laing, Alison; Howden, Mark; Little, Stephen
2014-08-01
The Murray dairy region produces approximately 1.85 billion litres of milk each year, representing about 20 % of Australia's total annual milk production. An ongoing production challenge in this region is the management of the impacts of heat stress during spring and summer. An increase in the frequency and severity of extreme temperature events due to climate change may result in additional heat stress and production losses. This paper assesses the changing nature of heat stress now, and into the future, using historical data and climate change projections for the region using the temperature humidity index (THI). Projected temperature and relative humidity changes from two global climate models (GCMs), CSIRO MK3.5 and CCR-MIROC-H, have been used to calculate THI values for 2025 and 2050, and summarized as mean occurrence of, and mean length of consecutive high heat stress periods. The future climate scenarios explored show that by 2025 an additional 12-15 days (compared to 1971 to 2000 baseline data) of moderate to severe heat stress are likely across much of the study region. By 2050, larger increases in severity and occurrence of heat stress are likely (i.e. an additional 31-42 moderate to severe heat stress days compared with baseline data). This increasing trend will have a negative impact on milk production among dairy cattle in the region. The results from this study provide useful insights on the trends in THI in the region. Dairy farmers and the dairy industry could use these results to devise and prioritise adaptation options to deal with projected increases in heat stress frequency and severity.
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Herbert, A G; Le Gros, G S; Bidawid, S; Watson, J D
1984-01-01
Cytotoxic effector cell populations in murine spleen can be characterized by the phenotype of the cytotoxic cells or the nature of target cells. Lytic events can be antigen-specific, MHC-restricted and clonal, or target cell-specific but apparently non-MHC-restricted. Two cytotoxic effectors of this latter category are spontaneous and natural killers. Normal spleen cells from (BALB/c X DBA/2J)F1 mice (CDF1) cultured without added antigen develop a population of Thy-1+ spontaneous cytotoxic lymphocytes (SCTL) that lyse the DBA/2J mastocytoma P815, as well as the BALB/c-derived plasmacytomas MOPC-11 and SP2/0. Cold target competition experiments reveal the BALB/c-derived plasmacytomas MOPC-11, SP2/0, J558 and the A strain-derived T cell lymphoma YAC-1, but not normal lymphoblasts, block the lysis of P815 target cells. Thus, while these tumour cells appear to express common antigens which are recognized by SCTL cells, plasmacytomas such as J558 are not susceptible to lysis by SCTL. The relationship of SCTL to natural killer (NK) cells was examined. In-vivo treatment of mice with monoclonal anti-Thy-1 antibody leads to a rapid loss of SCTL and precursors from the spleen, but there is a concomitant increase in NK cell activity. PMID:6607213
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Selective Manipulation of Neural Circuits.
Park, Hong Geun; Carmel, Jason B
2016-04-01
Unraveling the complex network of neural circuits that form the nervous system demands tools that can manipulate specific circuits. The recent evolution of genetic tools to target neural circuits allows an unprecedented precision in elucidating their function. Here we describe two general approaches for achieving circuit specificity. The first uses the genetic identity of a cell, such as a transcription factor unique to a circuit, to drive expression of a molecule that can manipulate cell function. The second uses the spatial connectivity of a circuit to achieve specificity: one genetic element is introduced at the origin of a circuit and the other at its termination. When the two genetic elements combine within a neuron, they can alter its function. These two general approaches can be combined to allow manipulation of neurons with a specific genetic identity by introducing a regulatory gene into the origin or termination of the circuit. We consider the advantages and disadvantages of both these general approaches with regard to specificity and efficacy of the manipulations. We also review the genetic techniques that allow gain- and loss-of-function within specific neural circuits. These approaches introduce light-sensitive channels (optogenetic) or drug sensitive channels (chemogenetic) into neurons that form specific circuits. We compare these tools with others developed for circuit-specific manipulation and describe the advantages of each. Finally, we discuss how these tools might be applied for identification of the neural circuits that mediate behavior and for repair of neural connections.
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McKernan, L N; Largen, M T
1984-09-01
The cloned monocyte/macrophage cell line RAW 264.7 was previously shown to produce thymocyte mitogenic and co-mitogenic activity that eluted from a Sephadex G-75 column not only at approximately 16,000 daltons, the m.w. described for interleukin 1 (IL 1), but also at 30,000 to 40,000 daltons. The studies reported here indicate that the 30,000 to 40,000 dalton molecule has thymic differentiating activity. Thymocytes from A/J mice were fractionated on discontinuous BSA gradients, which yielded populations of cells enriched for immature and mature cells. The cells found at the interface between 35 and 29% BSA (band 1 cells), which are the most immature, were cultured for 48 hr with highly purified IL 1, with the 30,000 to 40,000 dalton form of thymocyte co-mitogenic activity obtained after Sephadex G-75 chromatography and chromatofocusing chromatography, or with media alone. The surface antigens TL-3, H-2Kk, Thy-1.2, Lyt-1, and Lyt-2 were examined by immunofluorescence. It was found that the highly purified 30,000 to 40,000 dalton species of co-mitogenic activity induced a significant increase in the content of surface H-2Kk, a decrease in TL-3, and a very small decrease in Thy-1.2 on the cell surface, whereas IL 1 was not capable of inducing a change in these surface antigens. There was no change in Lyt-1 on the surface of band 1 thymocytes after incubation with either IL 1 or the 30,000 to 40,000 dalton species. The 30,000 to 40,000 dalton species caused a significant decrease in the percentage of cells staining positive for Lyt-2, whereas IL 1 caused a smaller but significant decrease in Lyt-2. These changes in the surface markers TL-3, H-2Kk, and Thy-1.2 are consistent with changes that occur during thymocyte differentiation. It was also observed that the proliferative response to the 30,000 to 40,000 dalton form and IL 1 increased with increasing functional maturity of each band of thymocytes when used in the thymocyte mitogenic assay. However, only the 30,000 to 40,000 dalton form was capable of inducing a proliferative response in the immature band 1 thymocytes in the thymocyte co-mitogenic assay. These results indicate that the RAW 264.7 cells produce a factor that has, in addition to thymocyte co-mitogenic activity, thymocyte differentiation activity, and this factor is distinct from IL 1.
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Martino Adami, Pamela V; Galeano, Pablo; Wallinger, Marina L; Quijano, Celia; Rabossi, Alejandro; Pagano, Eleonora S; Olivar, Natividad; Reyes Toso, Carlos; Cardinali, Daniel; Brusco, Luis I; Do Carmo, Sonia; Radi, Rafael; Gevorkian, Goar; Castaño, Eduardo M; Cuello, A Claudio; Morelli, Laura
2017-03-01
Diet is a modifiable risk factor for Alzheimer's disease (AD), but the mechanisms linking alterations in peripheral metabolism and cognition remain unclear. Since it is especially difficult to study long-term effects of high-energy diet in individuals at risk for AD, we addressed this question by using the McGill-R-Thy1-APP transgenic rat model (Tg(+/-)) that mimics presymptomatic AD. Wild-type and Tg(+/-) rats were exposed during 6months to a standard diet or a Western diet (WD), high in saturated fat and sugar. Results from peripheral and hippocampal biochemical analysis and in situ respirometry showed that WD induced a metabolic syndrome and decreased presynaptic bioenergetic parameters without alterations in hippocampal insulin signaling or lipid composition. Cognitive tests, ELISA multiplex, Western blot, immunohistochemistry and RT-qPCR indicated that WD worsened cognition in Tg(+/-) rats, increased hippocampal levels of monomeric Aβ isoforms and oligomeric species, promoted deposits of N-Terminal pyroglutamate-Aβ (AβN3(pE)) in CA1 pyramidal neurons and interneurons, decreased transcript levels of genes involved in neuroprotective pathways such as Sirtuin-1 and increased nitrated proteins. Our results support the concept that in the presence of early Aβ pathology, diet-induced metabolic dysfunctions may contribute as a "second hit" to impair cognition. Noteworthy, such effect is not mediated by higher microglia activation or disruption of blood brain barrier. However, it may be attributed to increased amyloidogenic processing of amyloid precursor protein, generation of AβN3(pE) and dysregulation of pathways governed by Sirtuin-1. This evidence reinforces the implementation of prophylactic interventions in individuals at risk for AD. Copyright © 2016 Elsevier B.V. All rights reserved.
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Corneal neurotoxicity due to topical benzalkonium chloride.
Sarkar, Joy; Chaudhary, Shweta; Namavari, Abed; Ozturk, Okan; Chang, Jin-Hong; Yco, Lisette; Sonawane, Snehal; Khanolkar, Vishakha; Hallak, Joelle; Jain, Sandeep
2012-04-06
The aim of this study was to determine and characterize the effect of topical application of benzalkonium chloride (BAK) on corneal nerves in vivo and in vitro. Thy1-YFP+ neurofluorescent mouse eyes were treated topically with vehicle or BAK (0.01% or 0.1%). Wide-field stereofluorescence microscopy was performed to sequentially image the treated corneas in vivo every week for 4 weeks, and changes in stromal nerve fiber density (NFD) and aqueous tear production were determined. Whole-mount immunofluorescence staining of corneas was performed with antibodies to axonopathy marker SMI-32. Western immunoblot analyses were performed on trigeminal ganglion and corneal lysates to determine abundance of proteins associated with neurotoxicity and regeneration. Compartmental culture of trigeminal ganglion neurons was performed in Campenot devices to determine whether BAK affects neurite outgrowth. BAK-treated corneas exhibited significantly reduced NFD and aqueous tear production, and increased inflammatory cell infiltration and fluorescein staining at 1 week (P < 0.05). These changes were most significant after 0.1% BAK treatment. The extent of inflammatory cell infiltration in the cornea showed a significant negative correlation with NFD. Sequential in vivo imaging of corneas showed two forms of BAK-induced neurotoxicity: reversible neurotoxicity characterized by axonopathy and recovery, and irreversible neurotoxicity characterized by nerve degeneration and regeneration. Increased abundance of beta III tubulin in corneal lysates confirmed regeneration. A dose-related significant reduction in neurites occurred after BAK addition to compartmental cultures of dissociated trigeminal ganglion cells. Although both BAK doses (0.0001% and 0.001%) reduced nerve fiber length, the reduction was significantly more with the higher dose (P < 0.001). Topical application of BAK to the eye causes corneal neurotoxicity, inflammation, and reduced aqueous tear production.
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Corneal Neurotoxicity Due to Topical Benzalkonium Chloride
Sarkar, Joy; Chaudhary, Shweta; Namavari, Abed; Ozturk, Okan; Chang, Jin-Hong; Yco, Lisette; Sonawane, Snehal; Khanolkar, Vishakha; Hallak, Joelle; Jain, Sandeep
2012-01-01
Purpose. The aim of this study was to determine and characterize the effect of topical application of benzalkonium chloride (BAK) on corneal nerves in vivo and in vitro. Methods. Thy1-YFP+ neurofluorescent mouse eyes were treated topically with vehicle or BAK (0.01% or 0.1%). Wide-field stereofluorescence microscopy was performed to sequentially image the treated corneas in vivo every week for 4 weeks, and changes in stromal nerve fiber density (NFD) and aqueous tear production were determined. Whole-mount immunofluorescence staining of corneas was performed with antibodies to axonopathy marker SMI-32. Western immunoblot analyses were performed on trigeminal ganglion and corneal lysates to determine abundance of proteins associated with neurotoxicity and regeneration. Compartmental culture of trigeminal ganglion neurons was performed in Campenot devices to determine whether BAK affects neurite outgrowth. Results. BAK-treated corneas exhibited significantly reduced NFD and aqueous tear production, and increased inflammatory cell infiltration and fluorescein staining at 1 week (P < 0.05). These changes were most significant after 0.1% BAK treatment. The extent of inflammatory cell infiltration in the cornea showed a significant negative correlation with NFD. Sequential in vivo imaging of corneas showed two forms of BAK-induced neurotoxicity: reversible neurotoxicity characterized by axonopathy and recovery, and irreversible neurotoxicity characterized by nerve degeneration and regeneration. Increased abundance of beta III tubulin in corneal lysates confirmed regeneration. A dose-related significant reduction in neurites occurred after BAK addition to compartmental cultures of dissociated trigeminal ganglion cells. Although both BAK doses (0.0001% and 0.001%) reduced nerve fiber length, the reduction was significantly more with the higher dose (P < 0.001). Conclusion. Topical application of BAK to the eye causes corneal neurotoxicity, inflammation, and reduced aqueous tear production. PMID:22410563
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A Linear-Programming-Based Coal Preparation and Blending Technique.
1985-04-10
ti Im- m .. stri ctiol .]s. ore-,sent prob Ims f()r model f ormu I at ion howv r. Thi srecci: ic (rav ty, , can take on 2() va tlues ranqi n; from 1...icuqn i < to prhl emP at v"xessx walsh I it- Vt r 1 b1t, si t 1aI e’r Orp il an a1ccer. f ’ ii r"Itv ) t h 14 (d-,I ormu IAtior. vrfr 1n1(_ish -I iiis
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Spacecraft/Environment Interactions CAE Tool. Volume 1. Users Manual
1988-01-01
aproe fo ubii-: K Arr A /- - " ,-’- .71 . IX, Iy- ,, A A GAUDE’, LT COL, USAF FOR -1E COMltaNDE > (SignturJ; RITA C. SAGCALYN Division Director Thi...report hay boen reviwed by the ESD Public Affairs Office (PA) and is relea-able to the ,.rAtional Techni.a1 Infor-,ation Service (NTIS). Qualified...Parts. Other Options TlY- procedure described above is the simplest method , r ,. files, but excludes several options provided by . K \\11. 5000. If the
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Distribution of Mosquitoes in the Continental United States.
1980-12-01
taenia sapphirina perturbans Wyjeorn7yia smi thi No USAF installations DE LAWARE cantlens is canaderrsii riop."Itan.-3 ql-m cantatorl sit vest ris...atv’atus bahamensis Urano taenia evraticusl- iowii-4.6 ioiambdis sapphirina 4 , Latisquama mul rennafli Wyeomy ia nigri pat-pusl’ 7 haynei opis...septentrionalis Cul-ex lUrano taenia err’aticus2 , 3 lowii nigripal 9us’ 3 sapphirinal1 3*1 pilosus pipiens2,3 Kyeomyia J haynei A mitchzel ii 1
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Discrete Analog Processing for Tracking and Guidance Control
1980-11-01
be called the multi- sample algorithm, satisfies -4 67 tD (Da - d) 0 (4.2.2.3) Thus, this descent algorithm will determine a coefficient vector a... flJ -TI:-* IS; 7" rR(VI Dr TH~I ("vFP)ALLCj TT$ C_ F 2C OH Til TPACK I! NC SYS TE ! f- 1I3 cc cc *’I cc. CC snUpcF FIL1j: C~T 01C 0 (1 cc CC OEJCT F I LF
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Optical Computations for Image Bandwidth Compression.
1981-05-15
UnCl1 ed 3 ’ " * ~ SECURITY CLASSIFICA/ION OF TWIS PA E Doi&e, be,. Enteerdj’ . /j (I) RFRORT DOCUMENTATION PAGE EAPINSTRUCTIONS (I) ~tOT DCUMETATON...Bolling Air Force Base, D. C. 20332 . NME’/PG 14. MONITORING AGENCY NAME 8 AOORESS(/I dilletnt from Controlling Office) IS. SECURITY CLASS. (oI thi ,(il... SECURITY CL.ASS4FICATII_ TI.AGE(W7Ief Deja Entered)2 (3)25imulations.m f a!1 incoherent optical/ *1 video feedback processor.. * Unclassi fled
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Lereim, Ragnhild Reehorst; Oveland, Eystein; Xiao, Yichuan; Torkildsen, Øivind; Wergeland, Stig; Myhr, Kjell-Morten; Sun, Shao-Cong; Berven, Frode S
2016-09-01
The ubiquitin ligase Peli1 has previously been suggested as a potential treatment target in multiple sclerosis. In the multiple sclerosis disease model, experimental autoimmune encephalomyelitis, Peli1 knock-out led to less activated microglia and less inflammation in the central nervous system. Despite being important in microglia, Peli1 expression has also been detected in glial and neuronal cells. In the present study the overall brain proteomes of Peli1 knock-out mice and wild-type mice were compared prior to experimental autoimmune encephalomyelitis induction, at onset of the disease and at disease peak. Brain samples from the frontal hemisphere, peripheral from the extensive inflammatory foci, were analyzed using TMT-labeling of sample pools, and the discovered proteins were verified in individual mice using label-free proteomics. The greatest proteomic differences between Peli1 knock-out and wild-type mice were observed at the disease peak. In Peli1 knock-out a higher degree of antigen presentation, increased activity of adaptive and innate immune cells and alterations to proteins involved in iron metabolism were observed during experimental autoimmune encephalomyelitis. These results unravel global effects to the brain proteome when abrogating Peli1 expression, underlining the importance of Peli1 as a regulator of the immune response also peripheral to inflammatory foci during experimental autoimmune encephalomyelitis. The proteomics data is available in PRIDE with accession PXD003710.
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Spencer, Brian; Verma, Inder; Desplats, Paula; Morvinski, Dinorah; Rockenstein, Ed; Adame, Anthony; Masliah, Eliezer
2014-01-01
Alzheimer disease (AD) is characterized by widespread neurodegeneration throughout the association cortex and limbic system, deposition of amyloid-β peptide (Aβ) in the neuropil and around the blood vessels, and formation of neurofibrillary tangles. The endopeptidase neprilysin has been successfully used to reduce the accumulation of Aβ following intracranial viral vector delivery or ex vivo manipulated intracranial delivery. These therapies have relied on direct injections into the brain, whereas a clinically desirable therapy would involve i.v. infusion of a recombinant enzyme. We previously characterized a recombinant neprilysin that contained a 38-amino acid brain-targeting domain. Recombinant cell lines have been generated expressing this brain-targeted enzyme (ASN12). In this report, we characterize the ASN12 recombinant protein for pharmacology in a mouse as well as efficacy in two APPtg mouse models of AD. The recombinant ASN12 transited to the brain with a t½ of 24 h and accumulated to 1.7% of injected dose at 24 h following i.v. delivery. We examined pharmacodynamics in the tg2576 APPtg mouse with the prion promoter APP695 SWE mutation and in the Line41 mThy1 APP751 mutation mouse. Treatment of either APPtg mouse resulted in reduced Aβ, increased neuronal synapses, and improved learning and memory. In addition, the Line41 APPtg mice showed increased levels of C-terminal neuropeptide Y fragments and increased neurogenesis. These results suggest that the recombinant brain-targeted neprilysin, ASN12, may be an effective treatment for AD and warrant further investigation in clinical trials. PMID:24825898
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U. S. Army Medical Research and Development Technical Report
1978-09-30
skin preparation prior to cathe- ter insertion, thi- type of catheter care following insertion, and the duration of central venous catheterization . An...NO. 3 A study is now underway to determine the optimal techniques for central venous catheter dressing care. A specially designed dress- ing tray that...chronically maintained male rhesus monkeys with indwelling central venous catheters. Two specific problems were en- countered. (1) Available data
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USDA-ARS?s Scientific Manuscript database
Differences in the adrenal cortisol response of OmniGen-AF (OG) supplemented dairy cows to a corticotrophin releasing hormone (CRH) and vasopressin (VP) or an adrenocorticotropic hormone (ACTH) challenge when housed at different temperature-humidity indices (THI) were studied. Holstein cows (n=12; 1...
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USDA-ARS?s Scientific Manuscript database
Differences in the response of OmniGen-AF (OG) supplemented dairy cows to a corticotropin releasing hormone (CRH) and vasopressin (VP) or an adrenocorticotropic hormone (ACTH) challenge when housed at different temperature-humidity indices (THI) were studied. Holstein cows (n=12; 162±1 days in milk)...
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2012-04-22
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Matias, Mariana; Fortuna, Ana; Bicker, Joana; Silvestre, Samuel; Falcão, Amílcar; Alves, Gilberto
2017-11-15
The heterocycles dihydropyrimidin(thi)ones have been under intensive pharmacological research, but their pharmacokinetic properties remain almost unknown. Herein, fifty dihydropyrimidin(thi)ones were submitted to in vitro screening tests using parallel artificial membrane permeability assays (PAMPA) to evaluate their apparent permeability (Papp) through intestinal membrane and blood-brain barrier models, and cell-based assays to assess their interference on the efflux transporter P-glycoprotein (P-gp). Moreover, a set of kinetic and toxicological parameters was also estimated employing a new computational tool, the pkCSM. The in vitro results suggested that 82% of the test compounds have good intestinal permeability (Papp>1.1×10 -6 cm/s), and 66% of these are also expected to exhibit good permeability through blood-brain barrier (Papp>2.0×10 -6 cm/s); these findings are consistent with a high transport rate by passive transcellular pathway. In both PAMPA models, thiourea derivatives presented higher Papp values than the respective urea analogues, which were further corroborated by in silico predictions. The in vitro results also suggested a low extent of plasma protein binding for all compounds (Papp<1.0×10 -5 cm/s), and these findings were also supported by in silico data (unbound fraction ranging from 0.13 to 0.59). In addition, although approximately half of the compounds did not modulate P-gp at the tested concentrations (10 and 50μM), nine of them presented a trend to induce P-gp and particularly the chlorinated compounds exhibited a marked P-gp inhibition at 50μM. Furthermore, the in silico predictions suggested that half of the compounds have hepatotoxic potential. Overall, within this group of compounds, the thiourea derivatives containing an unsubstituted or a monosubstituted (NO 2 , CH 3 , OCH 3 ) phenyl ring attached to the position 4 of the dihydropyrimidine ring represented the most promising structures and should be considered in the subsequent studies of the development of new structurally related drug candidates. Copyright © 2017 Elsevier B.V. All rights reserved.
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Base Release and Modification in Solid-Phase DNA Exposed to Low-Energy Electrons.
Choofong, Surakarn; Cloutier, Pierre; Sanche, Léon; Wagner, J Richard
2016-11-01
Ionization generates a large number of secondary low-energy electrons (LEEs) with a most probable energy of approximately 10 eV, which can break DNA bonds by dissociative electron attachment (DEA) and lead to DNA damage. In this study, we investigated radiation damage to dry DNA induced by X rays (1.5 keV) alone on a glass substrate or X rays combined with extra LEEs (average energy of 5.8 eV) emitted from a tantalum (Ta) substrate under an atmosphere of N 2 and standard ambient conditions of temperature and pressure. The targets included calf-thymus DNA and double-stranded synthetic oligonucleotides. We developed analytical methods to measure the release of non-modified DNA bases from DNA and the formation of several base modifications by LC-MS/MS with isotopic dilution for precise quantification. The results show that the yield of non-modified bases as well as base modifications increase by 20-30% when DNA is deposited on a Ta substrate compared to that on a glass substrate. The order of base release (Gua > Ade > Thy ∼ Cyt) agrees well with several theoretical studies indicating that Gua is the most susceptible site toward sugar-phosphate cleavage. The formation of DNA damage by LEEs is explained by DEA leading to the release of non-modified bases involving the initial cleavage of N1-C1', C3'-O3' or C5'-O5' bonds. The yield of base modifications was lower than the release of non-modified bases. The main LEE-induced base modifications include 5,6-dihydrothymine (5,6-dHT), 5,6-dihydrouracil (5-dHU), 5-hydroxymethyluracil (5-HmU) and 5-formyluracil (5-ForU). The formation of base modifications by LEEs can be explained by DEA and cleavage of the C-H bond of the methyl group of Thy (giving 5-HmU and 5-ForU) and by secondary reactions of H atoms and hydride anions that are generated by primary LEE reactions followed by subsequent reaction with Cyt and Thy (giving 5,6-dHU and 5,6-dHT).
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NASA Astrophysics Data System (ADS)
Ho, Long-Phi; Chau, Nguyen-Xuan-Quang; Nguyen, Hong-Quan
2013-04-01
The Nhieu Loc - Thi Nghe basin is the most important administrative and business area of Ho Chi Minh City. Due to system complexity of the basin such as the increasing trend of rainfall intensity, (tidal) water level and land subsidence, the simulation of hydrological, hydraulic variables for flooding prediction seems rather not adequate in practical projects. The basin is still highly vulnerable despite of multi-million USD investment for urban drainage improvement projects since the last decade. In this paper, an integrated system analysis in both spatial and temporal aspects based on statistical, GIS and modelling approaches has been conducted in order to: (1) Analyse risks before and after projects, (2) Foresee water-related risk under uncertainties of unfavourable driving factors and (3) Develop a sustainable flood risk management strategy for the basin. The results show that given the framework of risk analysis and adaptive strategy, certain urban developing plans in the basin must be carefully revised and/or checked in order to reduce the highly unexpected loss in the future
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Cloverleaf microgyroscope with electrostatic alignment and tuning
NASA Technical Reports Server (NTRS)
Challoner, A. Dorian (Inventor); Gutierrez, Roman C. (Inventor); Tang, Tony K. (Inventor)
2007-01-01
A micro-gyroscope (10) having closed loop output operation by a control voltage (V.sub.ty), that is demodulated by a drive axis (x-axis) signal V.sub.thx of the sense electrodes (S1, S2), providing Coriolis torque rebalance to prevent displacement of the micro-gyroscope (10) on the output axis (y-axis) V.sub.thy.about.0. Closed loop drive axis torque, V.sub.tx maintains a constant drive axis amplitude signal, V.sub.thx. The present invention provides independent alignment and tuning of the micro-gyroscope by using separate electrodes and electrostatic bias voltages to adjust alignment and tuning. A quadrature amplitude signal, or cross-axis transfer function peak amplitude is used to detect misalignment that is corrected to zero by an electrostatic bias voltage adjustment. The cross-axis transfer function is either V.sub.thy/V.sub.ty or V.sub.tnx/V.sub.tx. A quadrature signal noise level, or difference in natural frequencies estimated from measurements of the transfer functions is used to detect residual mistuning, that is corrected to zero by a second electrostatic bias voltage adjustment.
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‘Leave Your Ego at the Door’: A Narrative Investigation into Effective Wingsuit Flying
Arijs, Cedric; Chroni, Stiliani; Brymer, Eric; Carless, David
2017-01-01
In recent years there has been a rapid growth in interest in extreme sports. For the most part research has focused on understanding motivations for participation in extreme sports and very little research has attempted to investigate the psychological structure of effective performance. Those few studies that have attempted to explore this issue have tested models designed for traditional sport on adventure sports. However, extreme sports are not the same as adventure sports or traditional sports. This study employed a narrative approach to investigate experiences of effective performance in the extreme sport of proximity wingsuit flying. An overarching theme we labeled ‘leave your ego at the door,’ emerged based on four sub-themes: (1) know thy self, (2) know thy skills, (3) know the environment now, and (4) tame the ‘inner animal.’ These themes are presented and discussed in relation to performance and discovery narratives identified within elite sport, thereby shedding light on how participants’ experiences of the extreme sport of proximity wingsuit flying differ from dominant stories within traditional sports. PMID:29204131
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Aubin, F; Alcalay, J; Dall'Acqua, F; Kripke, M L
1990-06-01
Although some psoralens are therapeutically active in the treatment of cutaneous hyperproliferative diseases when combined with UVA (320-400 nm) radiation, the toxic effects of these compounds have led physicians to seek new photochemotherapeutic agents. One such agent is 4,4',5'-trimethylazapsoralen (TMAP), a new bifunctional psoralen compound. We investigated the effects of repetitive treatments with TMAP plus UVA radiation on the number of dendritic immune cells in murine epidermis and on the induction of phototoxicity. Mice treated 3 times per week for 4 weeks with 129 microgram TMAP plus 10 kJ/m2 UVA radiation exhibited no gross or microscopic evidence of phototoxicity. During this treatment, the numbers of ATPase+, Ia+, and Thy-l+ dendritic epidermal cells were greatly reduced, and by the end of the treatment period, few dendritic immune cells could be detected. We conclude that morphological alterations of cutaneous immune cells can occur in the absence of overt phototoxicity, and that TMAP plus low-dose UVA radiation decreases the numbers of detectable Langerhans cells and Thy-1+ cells in murine skin.
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Beam and Plasma Physics Research
1990-06-01
La di~raDy in high power microwave computations and thi-ory and high energy plasma computations and theory. The HPM computations concentrated on...2.1 REPORT INDEX 7 2.2 TASK AREA 2: HIGH-POWER RF EMISSION AND CHARGED- PARTICLE BEAM PHYSICS COMPUTATION , MODELING AND THEORY 10 2.2.1 Subtask 02-01...Vulnerability of Space Assets 22 2.2.6 Subtask 02-06, Microwave Computer Program Enhancements 22 2.2.7 Subtask 02-07, High-Power Microwave Transvertron Design 23
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Effect of low level laser (LLL) on cochlear and vestibular inner ear including tinnitus
NASA Astrophysics Data System (ADS)
Rhee, Chung-Ku; Lim, Eun-Seok; Kim, Young-Saeng; Chung, Yong-Won; Jung, Jae-Yun; Chung, Phil-Sang
2006-02-01
Objectives: 1. To investigate preventive effect of LLL on gentamicin-induced vestibular ototoxicity. 2. To evaluate the effectiveness of lower level laser (LLL) in the treatment of tinnitus. Methods: 1. Twenty guinea pigs were divided into control and laser groups. Vestibular ototoxicity was induced by intratympanic injection of gentamicin into left ear. LLL was irradiated into left ear canal of animals in laser group. Vestibular function of the animals was evaluated with vertical and off-vertical axis rotation testing. 2. Forty patients with tinnitus were treated with ginkgo biloba orally and randomly divided into control and laser groups. The 20 patients of laser group received 80.4 J/cm2 of 830 nm laser, 3 times per week for 4 weeks, via transmeatal irradiation. Tinnitus was evaluated by visual analogue scale (VAS) and tinnitus handicap inventory (THI). Results: 1. Preventive effect of LLL to gentamicin induced vestibular ototoxicity was demonstrated by preventing reduction of gain in slow harmonic acceleration test and modulation in the off-vertical axis rotation test. 2. Eleven of 20 laser group patients have shown significant improvement in VAS and THI compared to those of the control group. Conclusions: 1. LLL therapy may have preventive effect to vestibular ototoxicity. 2. LLL therapy in combination with ginkgo biloba seems to be worth trying on patients with tinnitus.
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Noguchi, Yoshihiro; Takahashi, Masatoki; Ito, Taku; Fujikawa, Taro; Kawashima, Yoshiyuki; Kitamura, Ken
2016-10-01
To assess possible delayed recovery of the maximum speech discrimination score (SDS) when the audiometric threshold ceases to change. We retrospectively examined 20 patients with idiopathic sudden sensorineural hearing loss (ISSNHL) (gender: 9 males and 11 females, age: 24-71 years). The findings of pure-tone average (PTA), maximum SDS, auditory brainstem responses (ABRs), and tinnitus handicap inventory (THI) were compared among the three periods of 1-3 months, 6-8 months, and 11-13 months after ISSNHL onset. No significant differences were noted in PTA, whereas an increase of greater than or equal to 10% in maximum SDS was recognized in 9 patients (45%) from the period of 1-3 months to the period of 11-13 months. Four of the 9 patients showed 20% or more recovery of maximum SDS. No significant differences were observed in the interpeak latency difference between waves I and V and the interaural latency difference of wave V in ABRs, whereas an improvement in the THI grade was recognized in 11 patients (55%) from the period of 1-3 months to the period of 11-13 months. The present study suggested the incidence of maximum SDS restoration over 1 year after ISSNHL onset. These findings may be because of the effects of auditory plasticity via the central auditory pathway. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.
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Kapucu, Fikret E.; Välkki, Inkeri; Mikkonen, Jarno E.; Leone, Chiara; Lenk, Kerstin; Tanskanen, Jarno M. A.; Hyttinen, Jari A. K.
2016-01-01
Synchrony and asynchrony are essential aspects of the functioning of interconnected neuronal cells and networks. New information on neuronal synchronization can be expected to aid in understanding these systems. Synchronization provides insight in the functional connectivity and the spatial distribution of the information processing in the networks. Synchronization is generally studied with time domain analysis of neuronal events, or using direct frequency spectrum analysis, e.g., in specific frequency bands. However, these methods have their pitfalls. Thus, we have previously proposed a method to analyze temporal changes in the complexity of the frequency of signals originating from different network regions. The method is based on the correlation of time varying spectral entropies (SEs). SE assesses the regularity, or complexity, of a time series by quantifying the uniformity of the frequency spectrum distribution. It has been previously employed, e.g., in electroencephalogram analysis. Here, we revisit our correlated spectral entropy method (CorSE), providing evidence of its justification, usability, and benefits. Here, CorSE is assessed with simulations and in vitro microelectrode array (MEA) data. CorSE is first demonstrated with a specifically tailored toy simulation to illustrate how it can identify synchronized populations. To provide a form of validation, the method was tested with simulated data from integrate-and-fire model based computational neuronal networks. To demonstrate the analysis of real data, CorSE was applied on in vitro MEA data measured from rat cortical cell cultures, and the results were compared with three known event based synchronization measures. Finally, we show the usability by tracking the development of networks in dissociated mouse cortical cell cultures. The results show that temporal correlations in frequency spectrum distributions reflect the network relations of neuronal populations. In the simulated data, CorSE unraveled the synchronizations. With the real in vitro MEA data, CorSE produced biologically plausible results. Since CorSE analyses continuous data, it is not affected by possibly poor spike or other event detection quality. We conclude that CorSE can reveal neuronal network synchronization based on in vitro MEA field potential measurements. CorSE is expected to be equally applicable also in the analysis of corresponding in vivo and ex vivo data analysis. PMID:27803660
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Diniz, Luan Pereira; Tortelli, Vanessa; Matias, Isadora; Morgado, Juliana; Bérgamo Araujo, Ana Paula; Melo, Helen M; Seixas da Silva, Gisele S; Alves-Leon, Soniza V; de Souza, Jorge M; Ferreira, Sergio T; De Felice, Fernanda G; Gomes, Flávia Carvalho Alcantara
2017-07-12
Alzheimer's disease (AD) is characterized by progressive cognitive decline, increasingly attributed to neuronal dysfunction induced by amyloid-β oligomers (AβOs). Although the impact of AβOs on neurons has been extensively studied, only recently have the possible effects of AβOs on astrocytes begun to be investigated. Given the key roles of astrocytes in synapse formation, plasticity, and function, we sought to investigate the impact of AβOs on astrocytes, and to determine whether this impact is related to the deleterious actions of AβOs on synapses. We found that AβOs interact with astrocytes, cause astrocyte activation and trigger abnormal generation of reactive oxygen species, which is accompanied by impairment of astrocyte neuroprotective potential in vitro We further show that both murine and human astrocyte conditioned media (CM) increase synapse density, reduce AβOs binding, and prevent AβO-induced synapse loss in cultured hippocampal neurons. Both a neutralizing anti-transforming growth factor-β1 (TGF-β1) antibody and siRNA-mediated knockdown of TGF-β1, previously identified as an important synaptogenic factor secreted by astrocytes, abrogated the protective action of astrocyte CM against AβO-induced synapse loss. Notably, TGF-β1 prevented hippocampal dendritic spine loss and memory impairment in mice that received an intracerebroventricular infusion of AβOs. Results suggest that astrocyte-derived TGF-β1 is part of an endogenous mechanism that protects synapses against AβOs. By demonstrating that AβOs decrease astrocyte ability to protect synapses, our results unravel a new mechanism underlying the synaptotoxic action of AβOs in AD. SIGNIFICANCE STATEMENT Alzheimer's disease is characterized by progressive cognitive decline, mainly attributed to synaptotoxicity of the amyloid-β oligomers (AβOs). Here, we investigated the impact of AβOs in astrocytes, a less known subject. We show that astrocytes prevent synapse loss induced by AβOs, via production of transforming growth factor-β1 (TGF-β1). We found that AβOs trigger morphological and functional alterations in astrocytes, and impair their neuroprotective potential. Notably, TGF-β1 reduced hippocampal dendritic spine loss and memory impairment in mice that received intracerebroventricular infusions of AβOs. Our results describe a new mechanism underlying the toxicity of AβOs and indicate novel therapeutic targets for Alzheimer's disease, mainly focused on TGF-β1 and astrocytes. Copyright © 2017 the authors 0270-6474/17/376798-13$15.00/0.
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Tsutsui, Yusuke; Schweicher, Guillaume; Chattopadhyay, Basab; Sakurai, Tsuneaki; Arlin, Jean-Baptiste; Ruzié, Christian; Aliev, Almaz; Ciesielski, Artur; Colella, Silvia; Kennedy, Alan R; Lemaur, Vincent; Olivier, Yoann; Hadji, Rachid; Sanguinet, Lionel; Castet, Frédéric; Osella, Silvio; Dudenko, Dmytro; Beljonne, David; Cornil, Jérôme; Samorì, Paolo; Seki, Shu; Geerts, Yves H
2016-09-01
The structural and electronic properties of four isomers of didodecyl[1]-benzothieno[3,2-b][1]benzothiophene (C12-BTBT) have been investigated. Results show the strong impact of the molecular packing on charge carrier transport and electronic polarization properties. Field-induced time-resolved microwave conductivity measurements unravel an unprecedented high average interfacial mobility of 170 cm(2) V(-1) s(-1) for the 2,7-isomer, holding great promise for the field of organic electronics. © 2016 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Oestrous behaviour of Holstein cows during cooler and hotter tropical seasons.
Rodtian, P; King, G; Subrod, S; Pongpiachan, P
1996-12-02
Seasonal effects on post-partum ovarian activity, duration and intensity of sexual behaviour were determined for Holstein dairy cattle imported from a temperate climate into a tropical region. Animals were observed continuously during the cooler (temperature-humidity index (THI) < 25) and hotter (THI > 25) seasons for 2 years. They were restricted to a cement footing in the hotter season observation period in Year 1, but had access to both concrete and dirt footing during all other seasons. Sequential milk progesterone profiles provided an indication of when follicular phases occurred, and recorded sexual behaviour was compared with these to determine if oestrous signs accompanied ovulations. Most cows had normal ovarian cycles and ovulated regularly during both seasons, but quiet ovulations occurred with greater frequency during the hotter times of the year (P < 0.05). Demonstrations of sexual behaviour were affected by choice of footing rather than season. The actual time when cows stood passively and allowed herdmates to complete mounting ranged from 5.1 +/- 0.7 to 5.8 +/- 1 h with access to exercise yards and cement or dirt footing, but declined to only 1.3 +/- 1.1 h when animals were confined to cement (P < 0.05). Similarly, the total duration of oestrus and mean number of interactions were significantly (P < 0.05) reduced during the observation period conducted with cows confined to concrete footing. These findings further emphasize that the duration of oestrus in dairy cows in considerably shorter than the commonly quoted 18 h.