Tuning the ion selectivity of two-pore channels

DOE Office of Scientific and Technical Information (OSTI.GOV)

Guo, Jiangtao; Zeng, Weizhong; Jiang, Youxing

Organellar two-pore channels (TPCs) contain two copies of a Shaker-like six-transmembrane (6-TM) domain in each subunit and are ubiquitously expressed in plants and animals. Interestingly, plant and animal TPCs share high sequence similarity in the filter region, yet exhibit drastically different ion selectivity. Plant TPC1 functions as a nonselective cation channel on the vacuole membrane, whereas mammalian TPC channels have been shown to be endo/lysosomal Na+-selective or Ca2+-release channels. In this study, we performed systematic characterization of the ion selectivity of TPC1 from Arabidopsis thaliana (AtTPC1) and compared its selectivity with the selectivity of human TPC2 (HsTPC2). We demonstrate thatmore » AtTPC1 is selective for Ca2+ over Na+, but nonselective among monovalent cations (Li+, Na+, and K+). Our results also confirm that HsTPC2 is a Na+-selective channel activated by phosphatidylinositol 3,5-bisphosphate. Guided by our recent structure of AtTPC1, we converted AtTPC1 to a Na+-selective channel by mimicking the selectivity filter of HsTPC2 and identified key residues in the TPC filters that differentiate the selectivity between AtTPC1 and HsTPC2. Furthermore, the structure of the Na+-selective AtTPC1 mutant elucidates the structural basis for Na+ selectivity in mammalian TPCs.« less

On the cellular site of two-pore channel TPC1 action in the Poaceae.

PubMed

Dadacz-Narloch, Beata; Kimura, Sachie; Kurusu, Takamitsu; Farmer, Edward E; Becker, Dirk; Kuchitsu, Kazuyuki; Hedrich, Rainer

2013-11-01

The slow vacuolar (SV) channel has been characterized in different dicots by patch-clamp recordings. This channel represents the major cation conductance of the largest organelle in most plant cells. Studies with the tpc1-2 mutant of the model dicot plant Arabidopsis thaliana identified the SV channel as the product of the TPC1 gene. By contrast, research on rice and wheat TPC1 suggested that the monocot gene encodes a plasma membrane calcium-permeable channel. To explore the site of action of grass TPC1 channels, we expressed OsTPC1 from rice (Oryza sativa) and TaTPC1 from wheat (Triticum aestivum) in the background of the Arabidopsis tpc1-2 mutant. Cross-species tpc1 complementation and patch-clamping of vacuoles using Arabidopsis and rice tpc1 null mutants documented that both monocot TPC1 genes were capable of rescuing the SV channel deficit. Vacuoles from wild-type rice but not the tpc1 loss-of-function mutant harbor SV channels exhibiting the hallmark properties of dicot TPC1/SV channels. When expressed in human embryonic kidney (HEK293) cells OsTPC1 was targeted to Lysotracker-Red-positive organelles. The finding that the rice TPC1, just like those from the model plant Arabidopsis and even animal cells, is localized and active in lyso-vacuolar membranes associates this cation channel species with endomembrane function. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.

The electronics and data acquisition system for the DarkSide-50 veto detectors

DOE PAGES

Agnes, P.; Agostino, L.; Albuquerque, I. F. M.; ...

2016-12-01

DarkSide-50 is a detector for dark matter candidates in the form of weakly interacting massive particles (WIMPs). It utilizes a liquid argon time projection chamber (LAr TPC) for the inner main detector. The TPC is surrounded by a liquid scintillator veto (LSV) and a water Cherenkov veto detector (WCV). The LSV and WCV, both instrumented with PMTs, act as the neutron and cosmogenic muon veto detectors for DarkSide-50. This paper describes the electronics and data acquisition system used for these two detectors.

Exploring the biophysical evidence that mammalian two‐pore channels are NAADP‐activated calcium‐permeable channels

PubMed Central

Reilly‐O'Donnell, Benedict; Sitsapesan, Rebecca

2016-01-01

Abstract Nicotinic acid adenine dinucleotide phosphate (NAADP) potently releases Ca2+ from acidic intracellular endolysosomal Ca2+ stores. It is widely accepted that two types of two‐pore channels, termed TPC1 and TPC2, are responsible for the NAADP‐mediated Ca2+ release but the underlying mechanisms regulating their gating appear to be different. For example, although both TPC1 and TPC2 are activated by NAADP, TPC1 appears to be additionally regulated by cytosolic Ca2+. Ion conduction and permeability also differ markedly. TPC1 and TPC2 are permeable to a range of cations although biophysical experiments suggest that TPC2 is slightly more selective for Ca2+ over K+ than TPC1 and hence capable of releasing greater quantities of Ca2+ from acidic stores. TPC1 is also permeable to H+ and therefore may play a role in regulating lysosomal and cytosolic pH, possibly creating localised acidic domains. The significantly different gating and ion conducting properties of TPC1 and TPC2 suggest that these two ion channels may play complementary physiological roles as Ca2+‐release channels of the endolysosomal system. PMID:26872338

Ionization Electron Signal Processing in Single Phase LArTPCs II. Data/Simulation Comparison and Performance in MicroBooNE

DOE Office of Scientific and Technical Information (OSTI.GOV)

Adams, C.; et al.

The single-phase liquid argon time projection chamber (LArTPC) provides a large amount of detailed information in the form of fine-grained drifted ionization charge from particle traces. To fully utilize this information, the deposited charge must be accurately extracted from the raw digitized waveforms via a robust signal processing chain. Enabled by the ultra-low noise levels associated with cryogenic electronics in the MicroBooNE detector, the precise extraction of ionization charge from the induction wire planes in a single-phase LArTPC is qualitatively demonstrated on MicroBooNE data with event display images, and quantitatively demonstrated via waveform-level and track-level metrics. Improved performance of inductionmore » plane calorimetry is demonstrated through the agreement of extracted ionization charge measurements across different wire planes for various event topologies. In addition to the comprehensive waveform-level comparison of data and simulation, a calibration of the cryogenic electronics response is presented and solutions to various MicroBooNE-specific TPC issues are discussed. This work presents an important improvement in LArTPC signal processing, the foundation of reconstruction and therefore physics analyses in MicroBooNE.« less

How to Advance TPC Benchmarks with Dependability Aspects

NASA Astrophysics Data System (ADS)

Almeida, Raquel; Poess, Meikel; Nambiar, Raghunath; Patil, Indira; Vieira, Marco

Transactional systems are the core of the information systems of most organizations. Although there is general acknowledgement that failures in these systems often entail significant impact both on the proceeds and reputation of companies, the benchmarks developed and managed by the Transaction Processing Performance Council (TPC) still maintain their focus on reporting bare performance. Each TPC benchmark has to pass a list of dependability-related tests (to verify ACID properties), but not all benchmarks require measuring their performances. While TPC-E measures the recovery time of some system failures, TPC-H and TPC-C only require functional correctness of such recovery. Consequently, systems used in TPC benchmarks are tuned mostly for performance. In this paper we argue that nowadays systems should be tuned for a more comprehensive suite of dependability tests, and that a dependability metric should be part of TPC benchmark publications. The paper discusses WHY and HOW this can be achieved. Two approaches are introduced and discussed: augmenting each TPC benchmark in a customized way, by extending each specification individually; and pursuing a more unified approach, defining a generic specification that could be adjoined to any TPC benchmark.

Endolysosomal two‐pore channels regulate autophagy in cardiomyocytes

PubMed Central

García‐Rúa, Vanessa; Feijóo‐Bandín, Sandra; Rodríguez‐Penas, Diego; Mosquera‐Leal, Ana; Abu‐Assi, Emad; Beiras, Andrés; María Seoane, Luisa; Lear, Pamela; Parrington, John; Portolés, Manuel; Roselló‐Lletí, Esther; Rivera, Miguel; Gualillo, Oreste; Parra, Valentina; Hill, Joseph A.; Rothermel, Beverly; González‐Juanatey, José Ramón

2016-01-01

Key points Two‐pore channels (TPCs) were identified as a novel family of endolysosome‐targeted calcium release channels gated by nicotinic acid adenine dinucleotide phosphate, as also as intracellular Na+ channels able to control endolysosomal fusion, a key process in autophagic flux.Autophagy, an evolutionarily ancient response to cellular stress, has been implicated in the pathogenesis of a wide range of cardiovascular pathologies, including heart failure.We report direct evidence indicating that TPCs are involved in regulating autophagy in cardiomyocytes, and that TPC knockout mice show alterations in the cardiac lysosomal system. TPC downregulation implies a decrease in the viability of cardiomyocytes under starvation conditions. In cardiac tissues from both humans and rats, TPC transcripts and protein levels were higher in females than in males, and correlated negatively with markers of autophagy.We conclude that the endolysosomal channels TPC1 and TPC2 are essential for appropriate basal and induced autophagic flux in cardiomyocytes, and also that they are differentially expressed in male and female hearts. Abstract Autophagy participates in physiological and pathological remodelling of the heart. The endolysosomal two‐pore channels (TPCs), TPC1 and TPC2, have been implicated in the regulation of autophagy. The present study aimed to investigate the role of TPC1 and TPC2 in basal and induced cardiac autophagic activity. In cultured cardiomyocytes, starvation induced a significant increase in TPC1 and TPC2 transcripts and protein levels that paralleled the increase in autophagy identified by increased LC3‐II and decreased p62 levels. Small interfering RNA depletion of TPC2 alone or together with TPC1 increased both LC3II and p62 levels under basal conditions and in response to serum starvation, suggesting that, under conditions of severe energy depletion (serum plus glucose starvation), changes in the autophagic flux (as assessed by use of bafilomycin A1) occurred either when TPC1 or TPC2 were downregulated. The knockdown of TPCs diminished cardiomyocyte viability under starvation and simulated ischaemia. Electron micrographs of hearts from TPC1/2 double knockout mice showed that cardiomyocytes contained large numbers of immature lysosomes with diameters significantly smaller than those of wild‐type mice. In cardiac tissues from humans and rats, TPC1 and TPC2 transcripts and protein levels were higher in females than in males. Furthermore, transcript levels of TPCs correlated negatively with p62 levels in heart tissues. TPC1 and TPC2 are essential for appropriate basal and induced autophagic flux in cardiomyocytes (i.e. there is a negative effect on cell viability under stress conditions in their absence) and they are differentially expressed in male and female human and murine hearts, where they correlate with markers of autophagy. PMID:26757341

Overview of TPC Benchmark E: The Next Generation of OLTP Benchmarks

NASA Astrophysics Data System (ADS)

Hogan, Trish

Set to replace the aging TPC-C, the TPC Benchmark E is the next generation OLTP benchmark, which more accurately models client database usage. TPC-E addresses the shortcomings of TPC-C. It has a much more complex workload, requires the use of RAID-protected storage, generates much less I/O, and is much cheaper and easier to set up, run, and audit. After a period of overlap, it is expected that TPC-E will become the de facto OLTP benchmark.

Two Pore Channel 2 (TPC2) Inhibits Autophagosomal-Lysosomal Fusion by Alkalinizing Lysosomal pH*

PubMed Central

Lu, Yingying; Hao, Bai-Xia; Graeff, Richard; Wong, Connie W. M.; Wu, Wu-Tian; Yue, Jianbo

2013-01-01

Autophagy is an evolutionarily conserved lysosomal degradation pathway, yet the underlying mechanisms remain poorly understood. Nicotinic acid adenine dinucleotide phosphate (NAADP), one of the most potent Ca2+ mobilizing messengers, elicits Ca2+ release from lysosomes via the two pore channel 2 (TPC2) in many cell types. Here we found that overexpression of TPC2 in HeLa or mouse embryonic stem cells inhibited autophagosomal-lysosomal fusion, thereby resulting in the accumulation of autophagosomes. Treatment of TPC2 expressing cells with a cell permeant-NAADP agonist, NAADP-AM, further induced autophagosome accumulation. On the other hand, TPC2 knockdown or treatment of cells with Ned-19, a NAADP antagonist, markedly decreased the accumulation of autophagosomes. TPC2-induced accumulation of autophagosomes was also markedly blocked by ATG5 knockdown. Interestingly, inhibiting mTOR activity failed to increase TPC2-induced autophagosome accumulation. Instead, we found that overexpression of TPC2 alkalinized lysosomal pH, and lysosomal re-acidification abolished TPC2-induced autophagosome accumulation. In addition, TPC2 overexpression had no effect on general endosomal-lysosomal degradation but prevented the recruitment of Rab-7 to autophagosomes. Taken together, our data demonstrate that TPC2/NAADP/Ca2+ signaling alkalinizes lysosomal pH to specifically inhibit the later stage of basal autophagy progression. PMID:23836916

Effect of thrombin preconditioning on migration of subventricular zone-derived cells after intracerebral hemorrhage in rats.

PubMed

Guan, Jingxia; Zhang, Shaofeng; Zhou, Qin; Yuan, Zhenhua; Lu, Zuneng

2016-09-01

To investigate the effect of thrombin preconditioning (TPC) on the intracerebral hemorrhage (ICH)-induced proliferation, migration, and function of subventriclular zone (SVZ) cells and to find new strategies that enhance endogenous neurogenesis after ICH. Male Sprague-Dawley rats were randomly divided into 3 groups (ICH, TPC, and control group). Rats of each group were randomly divided into 5 subgroups (3-d, 7-d, 14-d, 21-d, and 28-d subgroup). ICH was caused by intrastrial stereotactic administration of collagenase type IV. Brdu was used to label newborn SVZ cells. Organotypic brain slices were cultured to dynamically observe the migration of SVZ cells at living brain tissue. Migration of Dil-labeled SVZ cells in living brain slices was traced by time-lapse microscopy. To assess whether SVZ cells migrating to injured striatum had the ability to form synapses with other cells, brain slices from each group were double immunolabeled with Brdu and synapsin I. The number of Brdu-positive cells markedly increased in the ipsilateral SVZ and striatum 3 days after TPC, peaked at 14 days (P < 0.01), continued to 21 days, and then gradually decreased at 28 days with significant difference compared to the ICH group at each time point (P < 0.01). Migration of Dil-labeled SVZ cells in brain slices in each group was observed and imaged during a 12-h period. Dil-labeled SVZ cells in the TPC group were observed to migrate laterally toward striatum with time with a faster velocity compared to the ICH group (P < 0.01). Our study also demonstrated that TPC induced strong colocalization of Brdu and synapsin I in the ipsilateral striatum between 3 and 28 days after injury.TPC made colocalization of Brdu and synapsin I appear earlier and continue for a longer time compared to the ICH group. Our results demonstrated that TPC could promote proliferation, migration, and function of SVZ cells after ICH, which may provide a new idea for enhancing endogenous neurogenesis and developing new therapeutic strategies against ICH-induced brain injury.

Characterization of Two-Pore Channel 2 by Nuclear Membrane Electrophysiology

PubMed Central

Lee, Claire Shuk-Kwan; Tong, Benjamin Chun-Kit; Cheng, Cecily Wing-Hei; Hung, Harry Chun-Hin; Cheung, King-Ho

2016-01-01

Lysosomal calcium (Ca2+) release mediated by NAADP triggers signalling cascades that regulate many cellular processes. The identification of two-pore channel 2 (TPC2) as the NAADP receptor advances our understanding of lysosomal Ca2+ signalling, yet the lysosome is not amenable to traditional patch-clamp electrophysiology. Previous attempts to record TPC2 single-channel activity put TPC2 outside its native environment, which not reflect TPC2’s true physiological properties. To test the feasibility of using nuclear membrane electrophysiology for TPC2 channel characterization, we constructed a stable human TPC2-expressing DT40TKO cell line that lacks endogenous InsP3R and RyR (DT40TKO-hTPC2). Immunostaining revealed hTPC2 expression on the ER and nuclear envelope. Intracellular dialysis of NAADP into Fura-2-loaded DT40TKO-hTPC2 cells elicited cytosolic Ca2+ transients, suggesting that hTPC2 was functionally active. Using nuclear membrane electrophysiology, we detected a ~220 pS single-channel current activated by NAADP with K+ as the permeant ion. The detected single-channel recordings displayed a linear current-voltage relationship, were sensitive to Ned-19 inhibition, were biphasically regulated by NAADP concentration, and regulated by PKA phosphorylation. In summary, we developed a cell model for the characterization of the TPC2 channel and the nuclear membrane patch-clamp technique provided an alternative approach to rigorously investigate the electrophysiological properties of TPC2 with minimal manipulation. PMID:26838264

TPC2 controls pigmentation by regulating melanosome pH and size.

PubMed

Ambrosio, Andrea L; Boyle, Judith A; Aradi, Al E; Christian, Keith A; Di Pietro, Santiago M

2016-05-17

Melanin is responsible for pigmentation of skin and hair and is synthesized in a specialized organelle, the melanosome, in melanocytes. A genome-wide association study revealed that the two pore segment channel 2 (TPCN2) gene is strongly linked to pigmentation variations. TPCN2 encodes the two-pore channel 2 (TPC2) protein, a cation channel. Nevertheless, how TPC2 regulates pigmentation remains unknown. Here, we show that TPC2 is expressed in melanocytes and localizes to the melanosome-limiting membrane and, to a lesser extent, to endolysosomal compartments by confocal fluorescence and immunogold electron microscopy. Immunomagnetic isolation of TPC2-containing organelles confirmed its coresidence with melanosomal markers. TPCN2 knockout by means of clustered regularly interspaced short palindromic repeat/CRISPR-associated 9 gene editing elicited a dramatic increase in pigment content in MNT-1 melanocytic cells. This effect was rescued by transient expression of TPC2-GFP. Consistently, siRNA-mediated knockdown of TPC2 also caused a substantial increase in melanin content in both MNT-1 cells and primary human melanocytes. Using a newly developed genetically encoded pH sensor targeted to melanosomes, we determined that the melanosome lumen in TPC2-KO MNT-1 cells and primary melanocytes subjected to TPC2 knockdown is less acidic than in control cells. Fluorescence and electron microscopy analysis revealed that TPC2-KO MNT-1 cells have significantly larger melanosomes than control cells, but the number of organelles is unchanged. TPC2 likely regulates melanosomes pH and size by mediating Ca(2+) release from the organelle, which is decreased in TPC2-KO MNT-1 cells, as determined with the Ca(2+) sensor tyrosinase-GCaMP6. Thus, our data show that TPC2 regulates pigmentation through two fundamental determinants of melanosome function: pH and size.

Transaction Processing Performance Council (TPC): State of the Council 2010

NASA Astrophysics Data System (ADS)

Nambiar, Raghunath; Wakou, Nicholas; Carman, Forrest; Majdalany, Michael

The Transaction Processing Performance Council (TPC) is a non-profit corporation founded to define transaction processing and database benchmarks and to disseminate objective, verifiable performance data to the industry. Established in August 1988, the TPC has been integral in shaping the landscape of modern transaction processing and database benchmarks over the past twenty-two years. This paper provides an overview of the TPC's existing benchmark standards and specifications, introduces two new TPC benchmarks under development, and examines the TPC's active involvement in the early creation of additional future benchmarks.

A Total Pleural Covering for Lymphangioleiomyomatosis Prevents Pneumothorax Recurrence

PubMed Central

Kurihara, Masatoshi; Mizobuchi, Teruaki; Kataoka, Hideyuki; Sato, Teruhiko; Kumasaka, Toshio; Ebana, Hiroki; Yamanaka, Sumitaka; Endo, Reina; Miyahashira, Sumika; Shinya, Noriko; Seyama, Kuniaki

2016-01-01

Background Spontaneous pneumothorax is a major and frequently recurrent complication of lymphangioleiomyomatosis (LAM). Despite the customary use of pleurodesis to manage pnenumothorax, the recurrence rate remains high, and accompanying pleural adhesions cause serious bleeding during subsequent lung transplantation. Therefore, we have developed a technique of total pleural covering (TPC) for LAM to wrap the entire visceral pleura with sheets of oxidized regenerated cellulose (ORC) mesh, thereby reinforcing the affected visceral pleura and preventing recurrence. Methods Since January 2003, TPC has been applied during video-assisted thoracoscopic surgery for the treatment of LAM. The medical records of LAM patients who had TPC since that time and until August 2014 are reviewed. Results TPC was performed in 43 LAM patients (54 hemithoraces), 11 of whom required TPC bilaterally. Pneumothorax recurred in 14 hemithoraces (25.9%) from 11 patients (25.6%) after TPC. Kaplan-Meier estimates of recurrence-free hemithorax were 80.8% at 2.5 years, 71.7% at 5 years, 71.7% at 7.5 years, and 61.4% at 9 years. The recurrence-free probability was significantly better when 10 or more sheets of ORC mesh were utilized for TPC (P = 0.0018). TPC significantly reduced the frequency of pneumothorax: 0.544 ± 0.606 episode/month (mean ± SD) before TPC vs. 0.008 ± 0.019 after TPC (P<0.0001). Grade IIIa postoperative complications were found in 13 TPC surgeries (24.1%). Conclusions TPC successfully prevented the recurrence of pneumothorax in LAM, was minimally invasive and rarely caused restrictive ventilatory impairment. PMID:27658250

TPC2 controls pigmentation by regulating melanosome pH and size

PubMed Central

Ambrosio, Andrea L.; Boyle, Judith A.; Aradi, Al E.; Christian, Keith A.; Di Pietro, Santiago M.

2016-01-01

Melanin is responsible for pigmentation of skin and hair and is synthesized in a specialized organelle, the melanosome, in melanocytes. A genome-wide association study revealed that the two pore segment channel 2 (TPCN2) gene is strongly linked to pigmentation variations. TPCN2 encodes the two-pore channel 2 (TPC2) protein, a cation channel. Nevertheless, how TPC2 regulates pigmentation remains unknown. Here, we show that TPC2 is expressed in melanocytes and localizes to the melanosome-limiting membrane and, to a lesser extent, to endolysosomal compartments by confocal fluorescence and immunogold electron microscopy. Immunomagnetic isolation of TPC2-containing organelles confirmed its coresidence with melanosomal markers. TPCN2 knockout by means of clustered regularly interspaced short palindromic repeat/CRISPR-associated 9 gene editing elicited a dramatic increase in pigment content in MNT-1 melanocytic cells. This effect was rescued by transient expression of TPC2-GFP. Consistently, siRNA-mediated knockdown of TPC2 also caused a substantial increase in melanin content in both MNT-1 cells and primary human melanocytes. Using a newly developed genetically encoded pH sensor targeted to melanosomes, we determined that the melanosome lumen in TPC2-KO MNT-1 cells and primary melanocytes subjected to TPC2 knockdown is less acidic than in control cells. Fluorescence and electron microscopy analysis revealed that TPC2-KO MNT-1 cells have significantly larger melanosomes than control cells, but the number of organelles is unchanged. TPC2 likely regulates melanosomes pH and size by mediating Ca2+ release from the organelle, which is decreased in TPC2-KO MNT-1 cells, as determined with the Ca2+ sensor tyrosinase-GCaMP6. Thus, our data show that TPC2 regulates pigmentation through two fundamental determinants of melanosome function: pH and size. PMID:27140606

Isoform Specificity of Protein Kinase Cs in Synaptic Plasticity

ERIC Educational Resources Information Center

Sossin, Wayne S.

2007-01-01

Protein kinase Cs (PKCs) are implicated in many forms of synaptic plasticity. However, the specific isoform(s) of PKC that underlie(s) these events are often not known. We have used "Aplysia" as a model system in order to investigate the isoform specificity of PKC actions due to the presence of fewer isoforms and a large number of documented…

Enhancing the Anti-Solvatochromic Two-Photon Fluorescence for Cirrhosis Imaging by Forming a Hydrogen-Bond Network.

PubMed

Ren, Tian-Bing; Xu, Wang; Zhang, Qian-Ling; Zhang, Xing-Xing; Wen, Si-Yu; Yi, Hai-Bo; Yuan, Lin; Zhang, Xiao-Bing

2018-06-18

Two-photon imaging is an emerging tool for biomedical research and clinical diagnostics. Electron donor-acceptor (D-A) type molecules are the most widely employed two-photon scaffolds. However, current D-A type fluorophores suffer from solvatochromic quenching in aqueous biological samples. To address this issue, we devised a novel class of D-A type green fluorescent protein (GFP) chromophore analogues that form a hydrogen-bond network in water to improve the two-photon efficiency. Our design results in two-photon chalcone (TPC) dyes with 0.80 quantum yield and large two-photon action cross section (210 GM) in water. This strategy to form hydrogen bonds can be generalized to design two-photon materials with anti-solvatochromic fluorescence. To demonstrate the improved in vivo imaging, we designed a sulfide probe based on TPC dyes and monitored endogenous H 2 S generation and scavenging in the cirrhotic rat liver for the first time. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

MWPC prototyping and testing for STAR inner TPC upgrade

NASA Astrophysics Data System (ADS)

Shen, F.; Wang, S.; Yang, C.; Xu, Q.

2017-06-01

STAR experiment at the Relativistic Heavy Ion Collider (RHIC) is upgrading the inner sectors of the Time Projection Chamber (iTPC). The iTPC upgrade project will increase the segmentation on the inner pad plane from 13 to 40 pad rows and renew the inner sector wire chambers. The upgrade will expand the TPC's acceptance from |η|<=1.0 to |η|<=1.5. Furthermore, the detector will have better acceptance for tracks with low momentum, as well as better resolution in both momentum and dE/dx for tracks of all momenta. The enhanced measurement capabilities of STAR-iTPC upgrade are crucial to the physics program of the Phase II of Beam Energy Scan (BES-II) at RHIC during 2019-2020, in particular the QCD phase transition study. In this proceedings, I will discuss the iTPC MWPC module fabrication and testing results from the first full size iTPC MWPC pre-prototype made at Shandong University.

Two Isoforms of Dihydroxyacetone Phosphate Reductase from the Chloroplasts of Dunaliella tertiolecta.

PubMed

Gee, R.; Goyal, A.; Byerrum, R. U.; Tolbert, N. E.

1993-09-01

Three isoforms of dihydroxyacetone phosphate reductase in extracts from Dunaliella tertiolecta have been separated by a diethylaminoethyl cellulose column chromatography with a shallow NaCl gradient. The chloroplasts contained the two major isoforms, and the third, minor form was in the cytosol. The isoforms are unstable in the absence of glycerol and they are cold labile, but they may be partially reactivated at 35[deg]C. The first chloroplast form to elute from the DEAE cellulose column was the major form when the cells were grown on high NaCl and it has been referred to as the form for glycerol production for osmoregulation or "osmoregulator form." The second form increased in specific activity when inorganic phosphate was increased in the growth media to stimulate growth, and it has been given the designation for the form for glyceride synthesis, "glyceride form." The osmoregulator form was stimulated by NaCl added to the enzyme assay, but not by reduced Escherichia coli thioredoxin. The glyceride form had properties similar to the enzyme in leaf chloroplast, such as inhibition by NaCl and by fatty acyl-coenzyme A derivatives and some stimulation by dithiothreitol, uridine diphosphate galactose, cyti-dine diphosphate dipalmatoyl diglyceride, and reduced E. coli thioredoxin. Thus, Dunaliella chloroplasts have a salt-stimulated osmoregulatory form of dihydroxyacetone phosphate reductase, which seems to have a role in glycerol production, and an isoform, which may be involved in glyceride synthesis and which has properties similar to the enzyme in chloroplasts of higher plants.

Two Isoforms of Dihydroxyacetone Phosphate Reductase from the Chloroplasts of Dunaliella tertiolecta.

PubMed Central

Gee, R.; Goyal, A.; Byerrum, R. U.; Tolbert, N. E.

1993-01-01

Three isoforms of dihydroxyacetone phosphate reductase in extracts from Dunaliella tertiolecta have been separated by a diethylaminoethyl cellulose column chromatography with a shallow NaCl gradient. The chloroplasts contained the two major isoforms, and the third, minor form was in the cytosol. The isoforms are unstable in the absence of glycerol and they are cold labile, but they may be partially reactivated at 35[deg]C. The first chloroplast form to elute from the DEAE cellulose column was the major form when the cells were grown on high NaCl and it has been referred to as the form for glycerol production for osmoregulation or "osmoregulator form." The second form increased in specific activity when inorganic phosphate was increased in the growth media to stimulate growth, and it has been given the designation for the form for glyceride synthesis, "glyceride form." The osmoregulator form was stimulated by NaCl added to the enzyme assay, but not by reduced Escherichia coli thioredoxin. The glyceride form had properties similar to the enzyme in leaf chloroplast, such as inhibition by NaCl and by fatty acyl-coenzyme A derivatives and some stimulation by dithiothreitol, uridine diphosphate galactose, cyti-dine diphosphate dipalmatoyl diglyceride, and reduced E. coli thioredoxin. Thus, Dunaliella chloroplasts have a salt-stimulated osmoregulatory form of dihydroxyacetone phosphate reductase, which seems to have a role in glycerol production, and an isoform, which may be involved in glyceride synthesis and which has properties similar to the enzyme in chloroplasts of higher plants. PMID:12231930

Two Pore Channel 2 Differentially Modulates Neural Differentiation of Mouse Embryonic Stem Cells

PubMed Central

Zhang, Zhe-Hao; Lu, Ying-Ying; Yue, Jianbo

2013-01-01

Nicotinic acid adenine dinucleotide phosphate (NAADP) is an endogenous Ca2+ mobilizing nucleotide presented in various species. NAADP mobilizes Ca2+ from acidic organelles through two pore channel 2 (TPC2) in many cell types and it has been previously shown that NAADP can potently induce neuronal differentiation in PC12 cells. Here we examined the role of TPC2 signaling in the neural differentiation of mouse embryonic stem (ES) cells. We found that the expression of TPC2 was markedly decreased during the initial ES cell entry into neural progenitors, and the levels of TPC2 gradually rebounded during the late stages of neurogenesis. Correspondingly, TPC2 knockdown accelerated mouse ES cell differentiation into neural progenitors but inhibited these neural progenitors from committing to neurons. Overexpression of TPC2, on the other hand, inhibited mouse ES cell from entering the early neural lineage. Interestingly, TPC2 knockdown had no effect on the differentiation of astrocytes and oligodendrocytes of mouse ES cells. Taken together, our data indicate that TPC2 signaling plays a temporal and differential role in modulating the neural lineage entry of mouse ES cells, in that TPC2 signaling inhibits ES cell entry to early neural progenitors, but is required for late neuronal differentiation. PMID:23776607

Physicochemical properties and cell-based bioactivity of Pu'erh tea polysaccharide conjugates.

PubMed

Chen, Xiao-Qiang; Zhang, Zhi-Fa; Gao, Zhi-Ming; Huang, Yi; Wu, Zheng-Qi

2017-11-01

Polysaccharide conjugates were prepared from Pu'erh tea and fractionated by DEAE-cellulose DE-52 column chromatography to yield one unexplored polysaccharide-conjugate fraction termed TPC-P with a molecular weight of 251,200Da. DVS (dynamic vapour sorption) result discovered that the humidity condition of long-term preservation for TPC-P is below 70% RH. Although it contained proteins, TPC-P could not bind to the Coomassie Brilliant Blue dyes G250 and R250. The "shoulder-shaped" ultroviolet absorption peak in TPC-P UV-vis scanning spectum ascribe theabrownins that inevitably adsorbed the polysaccharide conjugate. Zeta potential results demonstrated TPC-P aqueous solution merely presented the negative charge properties of polysaccharides instead of acid-base property of its protein section, and had more stability in greater than pH 5.5. No precipitation or haze occurred in the three TPC-P/EGCG aqueous mixtures during their being stored for 12h. The phase separation was observed in aqueous mixtures of TPC-P and type B gelatin. TPC-P possessed the fine stability as a function of temperature heating and cooling between 0 and 55°C. It is proposed that some properties of the covalent binding protein of TPC-P were "shielded" by its polysaccharide chains. Copyright © 2017 Elsevier B.V. All rights reserved.

The effect of firefighter protective garments, self-contained breathing apparatus and exertion in the heat on postural sway.

PubMed

White, Scott C; Hostler, David

2017-08-01

Fire suppression wearing thermal protective clothing (TPC) and self-contained breathing apparatus (SCBA) challenges a firefighter's balance and may explain firefighter falls. Postural control based on force plate centre of pressure (COP) was compared for healthy subjects wearing TPC and SCBA before and after 20 min of heavy physical exertion in hot conditions. Baseline measures with and without TPC and SCBA (two different SCBA cylinder masses) were compared before and after exertion that included elements of fire suppression activities in an environmental chamber. COP excursion and variability increased with exertion for TPC and SCBA conditions compared to non-stressed conditions. The two different cylinder masses had no significant effect. Wearing TPC and SCBA when physically stressed in a hot environment increases postural sway and exacerbates postural control. Subjects compensated for the extra mass and adjusted to control postural sway with the addition of TPC and SCBA, but the stress protocol amplified these adjustments. Practitioner Summary: Firefighters wear thermal protective clothing (TPC) and self-contained breathing apparatus (SCBA) when heat-stressed and fatigued. Wearing TPC and SCBA was found to negatively impact balance when stressed, but not for non-stressed or two different sized SCBA tanks. Simulating fire-ground conditions wearing TPC and SCBA should be considered for improving balance.

Optimization of extraction parameters on the antioxidant properties of banana waste.

PubMed

Toh, Pui Yee; Leong, Fei Shan; Chang, Sui Kiat; Khoo, Hock Eng; Yim, Hip Seng

2016-01-01

Banana is grown worldwide and consumed as ripe fruit or used for culinary purposes. Peels form about 18-33% of the whole fruit and are discarded as a waste product. With a view to exploiting banana peel as a source of valuable compounds, this study was undertaken to evaluate the effect of different extraction parameters on the antioxidant activities of the industrial by-product of banana waste (peel). Influence of different extraction parameters such as types of solvent, percentages of solvent, and extraction times on total phenolic content (TPC) and antioxidant activity of mature and green peels of Pisang Abu (PA), Pisang Berangan (PB), and Pisang Mas (PM) were investigated. The best extraction parameters were initially selected based on different percentages of ethanol (0-100% v/v), extraction time (1-5 hr), and extraction temperature (25-60°C) for extraction of antioxidants in the banana peels. Total phenolic content (TPC) was evaluated using Folin-Ciocalteu reagent assay while antioxidant activities (AA) of banana peel were accessed by DPPH, ABTS, and β-carotene bleaching (BCB) assays at optimum extraction conditions. Based on different extraction solvents and percentages of solvents used, 70% and 90% of acetone had yielded the highest TPC for the mature and green PA peels, respectively; 90% of ethanol and methanol has yielded the highest TPC for the mature and green PB peels, respectively; while 90% ethanol for the mature and green PM peels. Similar extraction conditions were found for the antioxidant activities for the banana peel assessed using DPPH assay except for green PB peel, which 70% methanol had contributed to the highest AA. Highest TPC and AA were obtained by applying 4, 1, and 2 hrs extraction for the peels of PA, PB and PM, respectively. The best extraction conditions were also used for determination of AAs using ABTS and β-carotene bleaching assays. Therefore, the best extraction conditions used have given the highest TPC and AAs. By-products of banana (peel) can be considered as a potential source of antioxidants in food and pharmaceutical industry.

The two-pore channel TPC1 is required for efficient protein processing through early and recycling endosomes.

PubMed

Castonguay, Jan; Orth, Joachim H C; Müller, Thomas; Sleman, Faten; Grimm, Christian; Wahl-Schott, Christian; Biel, Martin; Mallmann, Robert Theodor; Bildl, Wolfgang; Schulte, Uwe; Klugbauer, Norbert

2017-08-30

Two-pore channels (TPCs) are localized in endo-lysosomal compartments and assumed to play an important role for vesicular fusion and endosomal trafficking. Recently, it has been shown that both TPC1 and 2 were required for host cell entry and pathogenicity of Ebola viruses. Here, we investigate the cellular function of TPC1 using protein toxins as model substrates for distinct endosomal processing routes. Toxin uptake and activation through early endosomes but not processing through other compartments were reduced in TPC1 knockout cells. Detailed co-localization studies with subcellular markers confirmed predominant localization of TPC1 to early and recycling endosomes. Proteomic analysis of native TPC1 channels finally identified direct interaction with a distinct set of syntaxins involved in fusion of intracellular vesicles. Together, our results demonstrate a general role of TPC1 for uptake and processing of proteins in early and recycling endosomes, likely by providing high local Ca 2+ concentrations required for SNARE-mediated vesicle fusion.

Optimized iterative decoding method for TPC coded CPM

NASA Astrophysics Data System (ADS)

Ma, Yanmin; Lai, Penghui; Wang, Shilian; Xie, Shunqin; Zhang, Wei

2018-05-01

Turbo Product Code (TPC) coded Continuous Phase Modulation (CPM) system (TPC-CPM) has been widely used in aeronautical telemetry and satellite communication. This paper mainly investigates the improvement and optimization on the TPC-CPM system. We first add the interleaver and deinterleaver to the TPC-CPM system, and then establish an iterative system to iteratively decode. However, the improved system has a poor convergence ability. To overcome this issue, we use the Extrinsic Information Transfer (EXIT) analysis to find the optimal factors for the system. The experiments show our method is efficient to improve the convergence performance.

Evolutionary and tissue-specific control of expression of multiple acyl-carrier protein isoforms in plants and bacteria.

PubMed

Battey, J F; Ohlrogge, J B

1990-02-01

We have examined the occurrence of multiple acyl-carrier protein (ACP), isoforms in evolutionarily diverse species of higher and lower plants. Isoforms were resolved by native polyacrylamide gel electrophoresis (PAGE), and were detected by Western blotting or fluorography of [(3)H]-palmitate-labelled ACPs. Multiple isoforms of ACP were found in leaf tissue of the monocotyledons Avena sativa and Hordeum vulgare and dicotyledons Arabidopsis thaliana, Cuphea wrightii, and Brassica napus. Lower vascular plants including the lycopod Selaginella krausseriana, the gymnosperms Ephedra sp. and Dioon edule, the ferns Davallia feejensis and Marsilea sp. and the most primitive known extant vascular plant, Psilotum nudum, were all found to have multiple ACP isoforms, as were the nonvascular liverworts, Lunularia sp. and Marchantia sp. and the moss, Polytrichum sp. Therefore, the development of ACP isoforms appears to have occurred early in plant evolution. However, we could detect only a single electrophoretic form of ACP in the unicellular algae Chlamydomonas reinhardtii and Dunaliella tertiolecta and the photosynthetic cyanobacteria Synechocystis strain 6803 and Agmnellum quadruplicatum. Thus, multiple forms of ACP do not occur in all photosynthetic organisms but may be associated with multicellular plants. We have also examined tissue specificity and light control over the expression of ACP isoforms. The relative abundance of multiple forms of ACP in leaf of Spinacia and Avena was altered very little by light. Rather, the different patterns of ACP isoforms were primarily dependent on the tissue type.

Transient Receptor Potential Mucolipin 1 (TRPML1) and Two-pore Channels Are Functionally Independent Organellar Ion Channels*

PubMed Central

Yamaguchi, Soichiro; Jha, Archana; Li, Qin; Soyombo, Abigail A.; Dickinson, George D.; Churamani, Dev; Brailoiu, Eugen; Patel, Sandip; Muallem, Shmuel

2011-01-01

NAADP is a potent second messenger that mobilizes Ca2+ from acidic organelles such as endosomes and lysosomes. The molecular basis for Ca2+ release by NAADP, however, is uncertain. TRP mucolipins (TRPMLs) and two-pore channels (TPCs) are Ca2+-permeable ion channels present within the endolysosomal system. Both have been proposed as targets for NAADP. In the present study, we probed possible physical and functional association of these ion channels. Exogenously expressed TRPML1 showed near complete colocalization with TPC2 and partial colocalization with TPC1. TRPML3 overlap with TPC2 was more modest. TRPML1 and to some extent TRPML3 co-immunoprecipitated with TPC2 but less so with TPC1. Current recording, however, showed that TPC1 and TPC2 did not affect the activity of wild-type TRPML1 or constitutively active TRPML1(V432P). N-terminally truncated TPC2 (TPC2delN), which is targeted to the plasma membrane, also failed to affect TRPML1 and TRPML1(V432P) channel function or TRPML1(V432P)-mediated Ca2+ influx. Whereas overexpression of TPCs enhanced NAADP-mediated Ca2+ signals, overexpression of TRPML1 did not, and the dominant negative TRPML1(D471K) was without affect on endogenous NAADP-mediated Ca2+ signals. Furthermore, the single channel properties of NAADP-activated TPC2delN were not affected by TRPML1. Finally, NAADP-evoked Ca2+ oscillations in pancreatic acinar cells were identical in wild-type and TRPML1−/− cells. We conclude that although TRPML1 and TPCs are present in the same complex, they function as two independent organellar ion channels and that TPCs, not TRPMLs, are the targets for NAADP. PMID:21540176

Sugars, ascorbic acid, total phenolic content and total antioxidant activity in passion fruit (Passiflora) cultivars.

PubMed

Devi Ramaiya, Shiamala; Bujang, Japar Sidik; Zakaria, Muta Harah; King, Wong Sing; Shaffiq Sahrir, Muhd Arif

2013-03-30

The levels of sugars, ascorbic acid, total phenolic content (TPC) and total antioxidant activity (TAA) were determined in fruit juices from seven passion fruit (Passiflora spp.) cultivars: P. edulis cultivars Purple, Frederick, Yellow, Pink, P. edulis f. flavicarpa, P. maliformis and P. quadrangularis (we also tested this cultivar's mesocarp). Purple and Yellow P. edulis had significantly higher total sugar, 142.85 ± 0.17 g kg⁻¹ and 139.69 ± 0.12 g kg⁻¹, respectively, than other cultivars. Glucose and fructose content were higher in juice from vine-ripened fruits of Purple, Frederick and Yellow P. edulis, P. quadrangularis and P. maliformis. Sucrose content was significantly higher in juice of non-vine-ripened fruits of P. edulis (Pink) and P. edulis f. flavicarpa. Ascorbic acid, TPC and TAA were significantly higher in vine-ripened Purple and Yellow P. edulis; ranges were 0.22-0.33 g kg⁻¹, 342.80-382.00 mg gallic acid equivalent L⁻¹ and 409.13-586.70 µmol Trolox L⁻¹, respectively. Based on principal component analysis (PCA) and cluster analysis, the main variables - °Brix, total sugar, glucose, fructose, ascorbic acid, TPC and TAA - formed the characteristics for the group comprising Purple and Yellow P. edulis. Glucose, fructose, sucrose, ascorbic acid, TAA and TPC were quantified in passion fruit juices. Variation of the above variables in juices of Passiflora depends on the cultivar and ripeness. © 2012 Society of Chemical Industry.

Redundant synthesis of a conidial polyketide by two distinct secondary metabolite clusters in Aspergillus fumigatus

PubMed Central

Throckmorton, Kurt; Lim, Fang Yun; Kontoyiannis, Dimitrios P.; Zheng, Weifa; Keller, Nancy P.

2016-01-01

Summary Filamentous fungi are renowned for the production of bioactive secondary metabolites. Typically, one distinct metabolite is generated from a specific secondary metabolite cluster. Here, we characterize the newly described trypacidin (tpc) cluster in the opportunistic human pathogen Aspergillus fumigatus. We find that this cluster as well as the previously characterized endocrocin (enc) cluster both contribute to the production of the spore metabolite endocrocin. Whereas trypacidin is eliminated when only tpc cluster genes are deleted, endocrocin production is only eliminated when both the tpc and enc non-reducing polyketide synthase-encoding genes, tpcC and encA, respectively, are deleted. EncC, an anthrone oxidase, converts the product released from EncA to endocrocin as a final product. In contrast, endocrocin synthesis by the tpc cluster likely results from incomplete catalysis by TpcK (a putative decarboxylase), as its deletion results in a nearly 10-fold increase in endocrocin production. We suggest endocrocin is likely a shunt product in all related non-reducing polyketide synthase clusters containing homologues of TpcK and TpcL (a putative anthrone oxidase), e.g. geodin and monodictyphenone. This finding represents an unusual example of two physically discrete secondary metabolite clusters generating the same natural product in one fungal species by distinct routes. PMID:26242966

Large Time Projection Chambers for Rare Event Detection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Heffner, M

The Time Projection Chamber (TPC) concept [add ref to TPC section] has been applied to many projects outside of particle physics and the accelerator based experiments where it was initially developed. TPCs in non-accelerator particle physics experiments are principally focused on rare event detection (e.g. neutrino and darkmater experiments) and the physics of these experiments can place dramatically different constraints on the TPC design (only extensions to the traditional TPCs are discussed here). The drift gas, or liquid, is usually the target or matter under observation and due to very low signal rates a TPC with the largest active massmore » is desired. The large mass complicates particle tracking of short and sometimes very low energy particles. Other special design issues include, efficient light collection, background rejection, internal triggering and optimal energy resolution. Backgrounds from gamma-rays and neutrons are significant design issues in the construction of these TPCs. They are generally placed deep underground to shield from cosmogenic particles and surrounded with shielding to reduce radiation from the local surroundings. The construction materials have to be carefully screened for radiopurity as they are in close contact with the active mass and can be a signification source of background events. The TPC excels in reducing this internal background because the mass inside the fieldcage forms one monolithic volume from which fiducial cuts can be made ex post facto to isolate quiet drift mass, and can be circulated and purified to a very high level. Self shielding in these large mass systems can be significant and the effect improves with density. The liquid phase TPC can obtain a high density at low pressure which results in very good self-shielding and compact installation with a lightweight containment. The down sides are the need for cryogenics, slower charge drift, tracks shorter than the typical electron diffusion, lower energy resolution (e.g. xenon) and limited charge readout options. Slower charge drift requires long electron lifetimes placing strict limits on the oxygen and other impurities with high electron affinity. A significant variation of the liquid phase TPC, that improves the charge readout, is the dual-phase TPC where a gas phase layer is formed above the liquid into which the drifting electrons are extracted and amplified, typically with electroluminescence. The successful transfer of electrons through the phase boundary requires careful control of its position and setting up an appropriate electric field. A high pressure gas phase TPC has no cryogenics and density is easily optimized for the signal, but a large heavy pressure vessel is required. Although shelf shielding is reduced, it can in some cases approach that of the liquid phase; in xenon at 50atm the density is about half that of water or about 1/6 of liquid xenon. A significant feature of high pressure xenon gas is the energy resolution. Below a density of about 0.5g/cc the intrinsic resolution is only a few times that of high purity germanium. A neutrino-less double beta decay (0{nu}2{beta}) TPC operated below this density limit could enjoy excellent energy resolution and maintain particle tracking for background rejection. An observable interaction with the TPC results in a charged particle that travels in the drift matter exciting and ionizing the atoms until the initial energy is converted into ionization, scintillation, or heat with relatively large fluctuations around a mean distribution. Rare event TPCs can be designed to detect scintillation light as well as charge to exploit the anti-correlation to improve energy resolution and/or signal to noise. An electric drift field separates the electrons and positive ions from the ionization although the separation is not complete and some electrons are captured, exciting atoms and releasing more light than the primary excitation alone. The average partition between the scintillation and ionization can be manipulated to increase the ionization (at a loss of scintillation) by a number of methods such as, increasing the strength of the electric field up to a saturation of the ionization yield, increasing the temperature to enhance the diffusion of the ionized electrons, and adding dopants such as triethylamine that can be photoionized by the scintillation photons releasing more ionization. Scintillation light is typically collected with photomultiplier tubes (PMTs) and avalanche photo diodes (APDs) although any fast (compared to the ionization drift speed) light collector capable of detecting the typically UV photons, maintaining high radiopurity and perhaps withstanding pressure would work. CCDs are slow and therefore only record 2 dimensions integrating over the time direction, some of which can be recovered with a few PMTs.« less

In vivo neuronal synthesis and axonal transport of Kunitz protease inhibitor (KPI)-containing forms of the amyloid precursor protein.

PubMed

Moya, K L; Confaloni, A M; Allinquant, B

1994-11-01

We have shown previously that the amyloid precursor protein (APP) is synthesized in retinal ganglion cells and is rapidly transported down the axons, and that different molecular weight forms of the precursor have different developmental time courses. Some APP isoforms contain a Kunitz protease inhibitor (KPI) domain, and APP that lacks the KPI domain is considered the predominant isoform in neurons. We now show that, among the various rapidly transported APPs, a 140-kDa isoform contains the KPI domain. This APP isoform is highly expressed in rapidly growing retinal axons, and it is also prominent in adult axon endings. This 140-kDa KPI-containing APP is highly sulfated compared with other axonally transported isoforms. These results show that APP with the KPI domain is a prominent isoform synthesized in neurons in vivo, and they suggest that the regulation of protease activity may be an important factor during the establishment of neuronal connections.

MWPC prototyping and performance test for the STAR inner TPC upgrade

NASA Astrophysics Data System (ADS)

Shen, Fuwang; Wang, Shuai; Kong, Fangang; Bai, Shiwei; Li, Changyu; Videbæk, Flemming; Xu, Zhangbu; Zhu, Chengguang; Xu, Qinghua; Yang, Chi

2018-07-01

A new prototype of STAR inner Time Projection Chamber (iTPC) MWPC sector has been fabricated and tested in an X-ray test system. The wire chamber built at Shandong University has a wire tension precision better than 6% and wire pitch precision better than 10 μm. The gas gain uniformity and energy resolution are measured to be better than 1% (RMS) and 20% (FWHM), respectively, using an 55Fe X-ray source. The iTPC upgrade project is to replace all 24 STAR TPC inner sectors as a crucial detector upgrade for the RHIC beam energy scan phase II program. The test results show that the constructed iTPC prototype meets all project requirements.

The ALICE TPC Upgrad

NASA Astrophysics Data System (ADS)

Castro, Andrew; Alice-Usa Collaboration; Alice-Tpc Collaboration

2017-09-01

The Time Projection Chamber (TPC) currently used for ALICE (A Large Ion Collider Experiment at CERN) is a gaseous tracking detector used to study both proton-proton and heavy-ion collisions at the Large Hadron Collider (LHC) In order to accommodate the higher luminosit collisions planned for the LHC Run-3 starting in 2021, the ALICE-TPC will undergo a major upgrade during the next LHC shut down. The TPC is limited to a read out of 1000 Hz in minimum bias events due to the intrinsic dead time associated with back ion flow in the multi wire proportional chambers (MWPC) in the TPC. The TPC upgrade will handle the increase in event readout to 50 kHz for heavy ion minimum bias triggered events expected with the Run-3 luminosity by switching the MWPCs to a stack of four Gaseous Electron Multiplier (GEM) foils. The GEM layers will combine different hole pitches to reduce the dead time while maintaining the current spatial and energy resolution of the existing TPC. Undertaking the upgrade of the TPC represents a massive endeavor in terms of design, production, construction, quality assurance, and installation, thus the upgrade is coordinated over a number of institutes worldwide. The talk will go over the physics motivation for the upgrade, the ALICE-USA contribution to the construction of Inner Read Out Chambers IROCs, and QA from the first chambers built in the U.S

Phosphorus content in three physical fractions of typical Chernozem

NASA Astrophysics Data System (ADS)

Kotelnikova, Anna; Egorova, Zoya; Sushkov, Nikolai; Matveeva, Natalia; Fastovets, Ilya; Rogova, Olga; Volkov, Dmitriy

2017-04-01

The widespread use of fertilizers makes it necessary to study not only the content but also the forms of occurrence of nutrients in soil, as well as the phase in which nutrients are transferred. These characteristics determine the availability of chemical elements for plants, but remain insufficiently studied. In this work we attempted to gain insight into the distribution of organo-mineral fractions in agriculturally used Chernozem from Voronezh (Russia) and the distribution of phosphorus - one of the most important nutrient elements - in this type of soil. We compared the distributions of phosphorus in physical fractions of the soil in 3 experimental groups: the control group (without fertilizers), the group fertilized with 1 dose of NPK, and the group fertilized with 2 doses of NPK. The soil was sampled during the period of treatment with fertilizers and during the period of aftereffect (4 years after the last application of fertilizers). In order to analyze organo-mineral fractions, we used size-density fractionation to separate the soil samples into three physical fractions: clay-associated fraction with particle size < 1μm (CF), light fraction with particle density < 2.0 g cm-3 (LF), and residual fraction > 2.0 g cm-3 (RF). Total phosphorus content (TPC) in the fractions was determined with Agilent 5100 ICP-AES spectrometer. To compare groups, simultaneous confidence intervals were computed from pooled variance estimators in ANOVA, and Fisher's LSD test was used. We showed that during the period of treatment with fertilizers LF increased proportionally to the dose of fertilizers, and a simultaneous reduction in RF was observed. During the period of aftereffect, the content of these fractions tended to the control value. The increase of LF may indicate increasing availability of nutrients, since this fraction is likely to participate in biological cycles. The analysis of TPC in fractions suggested that during the period of treatment with fertilizers most of phosphorus accumulates in CF. In the group with double dose of fertilizers TPC in CF was more than 1.5 times higher than in the control, while for LF the increase in TPC was not significant, and RF TPC was practically the same as in the control. Association of phosphorus predominantly with CF suggests that phosphorus was mainly adsorbed to the surface of clay particles rather than to organic components. Therefore, despite the increase in CF TPC as a consequence of treatment with fertilizers, the increase in availability of phosphorus is questionable. In the aftereffect period no significant differences in TPC were found. In conclusion, we showed that availability of fertilizers may be dependent on fractional composition of soil. Under our experimental conditions, phosphorus tended to bind predominantly to clay particles. However, in the aftereffect period, fractionation of TPC was similar to the control, indicating the need to further investigate the fate of phosphorus in soils.

Ligand Noninnocence in Iron Corroles: Insights from Optical and X-ray Absorption Spectroscopies and Electrochemical Redox Potentials

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ganguly, Sumit; Giles, Logan J.; Thomas, Kolle E.

Two new series of iron meso-tris(para-X-phenyl)corrole (TpXPC) complexes, Fe[TpXPC]Ph and Fe[TpXPC]Tol, in which X=CF 3, H, Me, and OMe, and Tol=p-methylphenyl (p-tolyl), have been synthesized, allowing a multitechnique electronic–structural comparison with the corresponding FeCl, FeNO, and Fe 2(μ-O) TpXPC derivatives. Optical spectroscopy revealed that the Soret maxima of the FePh and FeTol series are insensitive to the phenyl para substituent, consistent with the presumed innocence of the corrole ligand in these compounds. Accordingly, we may be increasingly confident in the ability of the substituent effect criterion to serve as a probe of corrole noninnocence. Furthermore, four complexes—Fe[TPC]Cl, Fe[TPC](NO), {Fe[TPC]} 2O,more » and Fe[TPC]Ph—were selected for a detailed XANES investigation of the question of ligand noninnocence. The intensity-weighted average energy (IWAE) positions were found to exhibit rather modest variations (0.8 eV over the series of corroles). The integrated Fe-K pre-edge intensities, on the other hand, vary considerably, with a 2.5 fold increase for Fe[TPC]Ph relative to Fe[TPC]Cl and Fe[TPC](NO). Given the approximately C 4v local symmetry of the Fe in all the complexes, the large increase in intensity for Fe[TPC]Ph may be attributed to a higher number of 3d holes, consistent with an expected Fe IV-like description, in contrast to Fe[TPC]Cl and Fe[TPC](NO), in which the Fe is thought to be Fe III-like. In conclusion, these results afford strong validation of XANES as a probe of ligand noninnocence in metallocorroles. Electrochemical redox potentials, on the other hand, were found not to afford a simple probe of ligand noninnocence in Fe corroles.« less

Ligand Noninnocence in Iron Corroles: Insights from Optical and X-ray Absorption Spectroscopies and Electrochemical Redox Potentials

DOE PAGES

Ganguly, Sumit; Giles, Logan J.; Thomas, Kolle E.; ...

2017-10-06

Two new series of iron meso-tris(para-X-phenyl)corrole (TpXPC) complexes, Fe[TpXPC]Ph and Fe[TpXPC]Tol, in which X=CF 3, H, Me, and OMe, and Tol=p-methylphenyl (p-tolyl), have been synthesized, allowing a multitechnique electronic–structural comparison with the corresponding FeCl, FeNO, and Fe 2(μ-O) TpXPC derivatives. Optical spectroscopy revealed that the Soret maxima of the FePh and FeTol series are insensitive to the phenyl para substituent, consistent with the presumed innocence of the corrole ligand in these compounds. Accordingly, we may be increasingly confident in the ability of the substituent effect criterion to serve as a probe of corrole noninnocence. Furthermore, four complexes—Fe[TPC]Cl, Fe[TPC](NO), {Fe[TPC]} 2O,more » and Fe[TPC]Ph—were selected for a detailed XANES investigation of the question of ligand noninnocence. The intensity-weighted average energy (IWAE) positions were found to exhibit rather modest variations (0.8 eV over the series of corroles). The integrated Fe-K pre-edge intensities, on the other hand, vary considerably, with a 2.5 fold increase for Fe[TPC]Ph relative to Fe[TPC]Cl and Fe[TPC](NO). Given the approximately C 4v local symmetry of the Fe in all the complexes, the large increase in intensity for Fe[TPC]Ph may be attributed to a higher number of 3d holes, consistent with an expected Fe IV-like description, in contrast to Fe[TPC]Cl and Fe[TPC](NO), in which the Fe is thought to be Fe III-like. In conclusion, these results afford strong validation of XANES as a probe of ligand noninnocence in metallocorroles. Electrochemical redox potentials, on the other hand, were found not to afford a simple probe of ligand noninnocence in Fe corroles.« less

Gait-Cycle-Driven Transmission Power Control Scheme for a Wireless Body Area Network.

PubMed

Zang, Weilin; Li, Ye

2018-05-01

In a wireless body area network (WBAN), walking movements can result in rapid channel fluctuations, which severely degrade the performance of transmission power control (TPC) schemes. On the other hand, these channel fluctuations are often periodic and are time-synchronized with the user's gait cycle, since they are all driven from the walking movements. In this paper, we propose a novel gait-cycle-driven transmission power control (G-TPC) for a WBAN. The proposed G-TPC scheme reinforces the existing TPC scheme by exploiting the periodic channel fluctuation in the walking scenario. In the proposed scheme, the user's gait cycle information acquired by an accelerometer is used as beacons for arranging the transmissions at the time points with the ideal channel state. The specific transmission power is then determined by using received signal strength indication (RSSI). An experiment was conducted to evaluate the energy efficiency and reliability of the proposed G-TPC based on a CC2420 platform. The results reveal that compared to the original RSSI/link-quality-indication-based TPC, G-TPC reduces energy consumption by 25% on the sensor node and reduce the packet loss rate by 65%.

Antioxidant activity and polyphenol content of green tea flavan-3-ols and oligomeric proanthocyanidins.

PubMed

Molan, Abdul L; De, Shampa; Meagher, Lucy

2009-09-01

The antioxidant activity and total phenolics content (TPC) of freshly prepared green tea extract (GTE) as affected by time, temperature and stirring were determined using the ferric reducing antioxidant power (FRAP) and Folin-Ciocalteu assays, respectively. Acetone-water fractions of GTE containing flavan-3-ols and oligomeric proanthocyanidins were evaluated at concentrations between 25 and 500 µg/ml. Increasing the extraction time from 3 min to 10 min resulted in a significant increase in both the FRAP values and TPC. Increasing the extraction time from 10 min to 30 min was without any significant effects on both FRAP and TPC values. Moreover, the FRAP values were correlated with the TPC. GTE fractions had widely different FRAP values that were well correlated with the TPC of the fraction. It was concluded that brewing conditions such as extraction temperature, period of extraction, ratio of tea leaves to extracting water, and stirring are important factors for determining the FRAP values and TPC in GTE. These factors should be taken into consideration during preparation for nutritional benefits during usual consumption of this beverage. Elevated FRAP and TPC values corresponded to those GTE fractions with a higher amount of phenolic compounds, which have stronger antioxidant activities.

Overview of the data analysis and new micro-pattern gas detector development for the Active Target Time Projection Chamber (AT-TPC) project.

NASA Astrophysics Data System (ADS)

Ayyad, Yassid; Mittig, Wolfgang; Bazin, Daniel; Cortesi, Marco

2017-07-01

The Active Target Time Projection Chamber (AT-TPC) project at the NSCL (National Superconducting Cyclotron Laboratory, Michigan State University) is a novel active target detector tailored for low-energy nuclear reactions in inverse kinematics with radioactive ion beams. The AT-TPC allows for a full three dimensional reconstruction of the reaction and provides high luminosity without degradation of resolution by the thickness of the target. Since all the particles (and also the reaction vertex) are tracked inside the detector, the AT-TPC has full 4π efficiency. The AT-TPC can operate under a magnetic field (2 T) that improves the identification of the particles and the energy resolution through the measurement of the magnetic rigidity. Another important characteristic of the AT-TPC is the high-gain operation achieved by the hybrid thick Gas Electron Multipliers (THGEM)-Micromegas pad plane, that allow operation also in pure elemental gas. These two features make the AT-TPC a unique high resolution spectrometer with full acceptance for nuclear physics reactions. This work presents an overview of the project, focused on the data analysis and the development of new micro-pattern gas detectors.

Relationship between total polar components and polycyclic aromatic hydrocarbons in fried edible oil.

PubMed

An, Ke-Jing; Liu, Yu-Lan; Liu, Hai-Lan

2017-09-01

Deep-fried dough sticks (a Chinese traditional breakfast) were fried individually in peanut, sunflower, rapeseed, rice bran, soybean and palm oil without any time lag for 32 h (64 batches fried, each for 30 min) and fried oil samples were obtained every 2 h. The frying-induced changes in the levels of total polar compounds (TPC) and polycyclic aromatic hydrocarbons (PAHs) were investigated by edible oil polar compounds (EOPC) fast separation chromatographic system and gas chromatography-mass spectrometry (GC-MS), respectively. The correlations were analysed of TPC with benzo[a]pyrene (BaP), TPC and PAH4 (benzo[a]anthracene, chrysene, benzo[b]fluoranthene and benzo[a]pyrene) as well as TPC with PAH16 (USEPA 16 PAHs). The results revealed that the levels of TPC and PAHs in fried oil considerably increased with frying time, and the type of oil affected their formation, which could inform the choice of oil for frying. The total BaP equivalents (∑BaPeq) concentrations in fresh oil and in oil whose TPC exceeded 27% were 2.14-13.48 and 5.78-10.80 μg kg -1 , respectively, which means that the carcinogenic potency of frying oil was more pronounced than that of fresh oil. In addition, the TPC concentration was significantly correlated with the concentrations of the sum of the 16 PAHs, PAH4 and BaP, so that the levels of PAHs could be predicted according to the levels of TPC in fried oil. In European standards, the rejection point for TPC in frying oil should be recalculated when considered PAHs. In all, the concentration of PAHs is a vital factor for ensuring the safety of frying oil.

Expression of c-Kit isoforms in multiple myeloma: differences in signaling and drug sensitivity.

PubMed

Montero, Juan Carlos; López-Pérez, Ricardo; San Miguel, Jesús F; Pandiella, Atanasio

2008-06-01

c-Kit is expressed in the plasma cells from 30% of patients with multiple myeloma. Two different isoforms of c-Kit, characterized by the presence or absence of the tetrapeptide sequence GNNK in the extracellular domain, have been described. However, their expression and function in myeloma cells are unknown. We explored the function and expression of these c-Kit isoforms in myeloma cells. Expression of c-Kit isoforms was investigated by reverse transcriptase polymerase chain reaction in fresh plasma cells from patients and cell lines. The function of these c-Kit isoforms was analyzed upon expression in myeloma cells. Signaling was investigated by western blotting using antibodies specific for activated forms of several signaling proteins. The impact of c-Kit on the action of drugs commonly used in the treatment of multiple myeloma was investigated by MTT proliferation assays. Fresh plasma cells from patients as well as myeloma cell lines expressed the two isoforms of c-Kit. Retroviral infection of myeloma cells with vectors that code for c-Kit-GNNK+ or c-Kit-GNNK- forms demonstrated differences in the kinetics of phosphorylation between these isoforms. Stem cell factor-induced activation of the GNNK- form was faster and more pronounced than that of the GNNK+ form, whose activation, however, lasted for longer. The c-Kit receptors weakly activated the Erk1/2 and Erk5 pathways. Both receptors, however, efficiently coupled to the PI3K/Akt pathway, and stimulated p70S6K activation. The latter was sensitive to the mTOR inhibitor, rapamycin. Studies of drug sensitivity indicated that cells expressing the GNNK- form were more resistant to the anti-myeloma action of bortezomib and melphalan. Our data indicate that c-Kit expression in multiple myeloma cells is functional, and coupled to survival pathways that may modulate cell death in response to therapeutic compounds used in the treatment of this disease.

A time projection chamber for high accuracy and precision fission cross-section measurements

DOE PAGES

Heffner, M.; Asner, D. M.; Baker, R. G.; ...

2014-05-22

The fission Time Projection Chamber (fissionTPC) is a compact (15 cm diameter) two-chamber MICROMEGAS TPC designed to make precision cross-section measurements of neutron-induced fission. The actinide targets are placed on the central cathode and irradiated with a neutron beam that passes axially through the TPC inducing fission in the target. The 4π acceptance for fission fragments and complete charged particle track reconstruction are powerful features of the fissionTPC which will be used to measure fission cross-sections and examine the associated systematic errors. This study provides a detailed description of the design requirements, the design solutions, and the initial performance ofmore » the fissionTPC.« less

MWPC prototyping and performance test for the STAR inner TPC upgrade

DOE PAGES

Shen, Fuwang; Wang, Shuai; Kong, Fangang; ...

2018-04-16

A new prototype of STAR inner Time Projection Chamber (iTPC) MWPC sector has been fabricated and tested in an X-ray test system. The wire chamber built at Shandong University has a wire tension precision better than 6% and wire pitch precision better than 10 μm. The gas gain uniformity and energy resolution are measured to be better than 1% (RMS) and 20% (FWHM), respectively, using an 55Fe X-ray source. The iTPC upgrade project is to replace all 24 STAR TPC inner sectors as a crucial detector upgrade for the RHIC beam energy scan phase II program. Furthermore, the test resultsmore » show that the constructed iTPC prototype meets all project requirements.« less

MWPC prototyping and performance test for the STAR inner TPC upgrade

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shen, Fuwang; Wang, Shuai; Kong, Fangang

A new prototype of STAR inner Time Projection Chamber (iTPC) MWPC sector has been fabricated and tested in an X-ray test system. The wire chamber built at Shandong University has a wire tension precision better than 6% and wire pitch precision better than 10 μm. The gas gain uniformity and energy resolution are measured to be better than 1% (RMS) and 20% (FWHM), respectively, using an 55Fe X-ray source. The iTPC upgrade project is to replace all 24 STAR TPC inner sectors as a crucial detector upgrade for the RHIC beam energy scan phase II program. Furthermore, the test resultsmore » show that the constructed iTPC prototype meets all project requirements.« less

Differential Regulation of Aromatase Isoforms and Tissue Responses to Environmental Chemicals in Fish

EPA Science Inventory

As in mammals, aromatase plays a basic role in fish reproduction. Unlike most mammals, with only one form of aromatase, fish have two distinct forms. One isoform, P450aromA, predominates in ovaries. Ovarian aromatase activity controls circulating levels of estrogens and is critic...

Stakeholder Participation in Freshwater Monitoring and Evaluation Programs: Applying Thresholds of Potential Concern within Environmental Flows

NASA Astrophysics Data System (ADS)

Conallin, John; McLoughlin, Craig A.; Campbell, Josh; Knight, Roger; Bright, Troy; Fisher, Ian

2018-03-01

The complex nature of freshwater systems provides challenges for incorporating evidence-based techniques into management. This paper investigates the potential of participatory evidence-based techniques to involve local stakeholders and make decisions based on different "knowledge" sources within adaptive management programs. It focuses on the application of thresholds of potential concern (TPC) within strategic adaptive management (SAM) for facilitating inclusive decision-making. The study is based on the case of the Edward-Wakool (E-W) "Fish and Flows" SAM project in the Murray-Darling River Basin, Australia. We demonstrate the application of TPCs for improving collaborative decision-making within the E-W, associated with environmental watering requirements, and other natural resource management programs such as fish stocking. The development of TPCs in the E-W fish and flows SAM project helped improve stakeholder involvement and understanding of the system, and also the effectiveness of the implemented management interventions. TPCs ultimately helped inform environmental flow management activities. The TPC process complemented monitoring that was already occurring in the system and provided a mechanism for linking formal and informal knowledge to form explicit and measurable endpoints from objectives. The TPC process faced challenges due to the perceived reduction in scientific rigor within initial TPC development and use. However, TPCs must remain tangible to managers and other stakeholders, in order to aid in the implementation of adaptive management. Once accepted by stakeholders, over time TPCs should be reviewed and refined in order to increase their scientific rigor, as new information is generated.

TPC2 mediates new mechanisms of platelet dense granule membrane dynamics through regulation of Ca2+ release

PubMed Central

Ambrosio, Andrea L.; Boyle, Judith A.; Di Pietro, Santiago M.

2015-01-01

Platelet dense granules (PDGs) are acidic calcium stores essential for normal hemostasis. They develop from late endosomal compartments upon receiving PDG-specific proteins through vesicular trafficking, but their maturation process is not well understood. Here we show that two-pore channel 2 (TPC2) is a component of the PDG membrane that regulates PDG luminal pH and the pool of releasable Ca2+. Using a genetically encoded Ca2+ biosensor and a pore mutant TPC2, we establish the function of TPC2 in Ca2+ release from PDGs and the formation of perigranular Ca2+ nanodomains. For the first time, Ca2+ spikes around PDGs—or any organelle of the endolysosome family—are visualized in real time and revealed to precisely mark organelle “kiss-and-run” events. Further, the presence of membranous tubules transiently connecting PDGs is revealed and shown to be dramatically enhanced by TPC2 in a mechanism that requires ion flux through TPC2. “Kiss-and-run” events and tubule connections mediate transfer of membrane proteins and luminal content between PDGs. The results show that PDGs use previously unknown mechanisms of membrane dynamics and content exchange that are regulated by TPC2. PMID:26202466

[Therapeutic Drug Monitoring of Valproic Acid in Children: A Prospective Study of The Effect of The Compliance and The Economic Level on the Trough Plasmatic Concentrations and Epileptic Seizures].

PubMed

Charfi, Rim; Lakhal, Mohamed; Klouz, Anis; Trabelsi, Sameh; Salouage, Issam

2015-01-01

Valproic acid (VA) is a widely used antiepileptic drug. Because of its pharmacokinetic variability and the influence of intrinsic and extrinsic factors such as the treatment compliance, VA therapeutic drug monitoring (TDM) is recommended in children. The aim of this study is to evaluate the effect of treatment compliance and the economic level on VA tough plasmatic concentration (TPC) and epileptic rhythm in children. A one-year prospective study (August 2008-August 2009) concerning children (age≤5 years) regularly treated by VA who had a VA TDM. So, 276 plasmatic samples from 238 children were collected. The children were divided in two groups as following: the group 1 (G1) presenting a good compliance and a reliable questioning and the group 2 (G2) presenting a bad compliance and a non reliable questioning. We evaluated the interindividual variability by correlating the TPC to the dose. Then, we divided the hole group in function of their economic levels (low-medium-high). Sex ratio male/female was 1.3. Median age was 5 years+/-3,9. The mean TPC was 62 µg/mL [0.12-131 µg/mL]. VA TPC were in the therapeutic range (TR) in 62%. Adverse drug reactions were noted in 4.2% of the children. G1 represented 70% of the children and G2, 30%. The TPC were in the TR in 67% of G1 and 51% of G2 (p=0.02). There was a significant difference between the TPC in G1 and G2 (p=0.02).There was no significative difference in the TPC in function of the economic levels. There was no correlation between TPC and the administered doses. The epileptic seizures were more spaced in children with therapeutic TPC than those with TPC in the TR (p=0.002) and in G1 than in G2 (p=0.03). Compliance should be appropriate in order to optimize the TDM rule. A good compliance and a therapeutic TPC allow a better control of epileptic seizures. © 2015 Société Française de Pharmacologie et de Thérapeutique.

Structural characterization of NRAS isoform 5

PubMed Central

Mal, Tapas K.; Yuan, Chunhua; Courtney, Nicholas B.; Patel, Mitra; Stiff, Andrew R.; Blachly, James; Walker, Christopher; Eisfeld, Ann‐Kathrin; de la Chapelle, Albert

2016-01-01

Abstract It was recently discovered that the NRAS isoform 5 (20 amino acids) is expressed in melanoma and results in a more aggressive cell phenotype. This novel isoform is responsible for increased phosphorylation of downstream targets such as AKT, MEK, and ERK as well as increased cellular proliferation. This structure report describes the NMR solution structure of NRAS isoform 5 to be used as a starting point to understand its biophysical interactions. The isoform is highly flexible in aqueous solution, but forms a helix‐turn‐coil structure in the presence of trifluoroethanol as determined by NMR and CD spectroscopy. PMID:26947772

Regulated Expression of a Calmodulin Isoform Alters Growth and Development in Potato

NASA Technical Reports Server (NTRS)

Poovaiah, B. W.; Takezawa, D.; An, G.; Han, T.-J.

1996-01-01

A transgene approach was taken to study the consequences of altered expression of a calmodutin iso-form on plant growth and development. Eight genomic clones of potato calmodulin (PCM 1 to 8) have been isolated and characterized. Among the potato calmodulin isoforms studied, PCM 1 differs from the other isoforms because of its unique amino acid substitutions. Transgenic potato plants were produced carrying sense construct of PCM 1 fused to the CAMV 35S promoter. Transgenic plants showing a moderate increase in PCM 1 MRNA exhibited strong apical dominance, produced elongated tubers, and were taller than the controls. Interestingly, the plants expressing the highest level of PCM 1 MRNA did not form underground tubers. Instead, these transgenic plants produced aerial tubers when allowed to grow for longer periods. The expression of different calmodulin isoforms (PCM 1, 5, 6, and 8) was studied in transgenic plants. Among the four potato calmodulin isoforms, only the expression of PCM 1 MRNA was altered in transgenic plants, while the expression of other isoforms was not significantly altered. Western analysis revealed increased PCM 1 protein in transgenic plants, indicating that the expression of both MRNA and protein are altered in transgenic plants. These results suggest that increasing the expression of PCM 1 alters growth and development in potato plants.

Lysosomal Two-pore Channel Subtype 2 (TPC2) Regulates Skeletal Muscle Autophagic Signaling*

PubMed Central

Lin, Pei-Hui; Duann, Pu; Komazaki, Shinji; Park, Ki Ho; Li, Haichang; Sun, Mingzhai; Sermersheim, Mathew; Gumpper, Kristyn; Parrington, John; Galione, Antony; Evans, A. Mark; Zhu, Michael X.; Ma, Jianjie

2015-01-01

Postnatal skeletal muscle mass is regulated by the balance between anabolic protein synthesis and catabolic protein degradation, and muscle atrophy occurs when protein homeostasis is disrupted. Autophagy has emerged as critical in clearing dysfunctional organelles and thus in regulating protein turnover. Here we show that endolysosomal two-pore channel subtype 2 (TPC2) contributes to autophagy signaling and protein homeostasis in skeletal muscle. Muscles derived from Tpcn2−/− mice exhibit an atrophic phenotype with exacerbated autophagy under starvation. Compared with wild types, animals lacking TPC2 demonstrated an enhanced autophagy flux characterized by increased accumulation of autophagosomes upon combined stress induction by starvation and colchicine treatment. In addition, deletion of TPC2 in muscle caused aberrant lysosomal pH homeostasis and reduced lysosomal protease activity. Association between mammalian target of rapamycin and TPC2 was detected in skeletal muscle, allowing for appropriate adjustments to cellular metabolic states and subsequent execution of autophagy. TPC2 therefore impacts mammalian target of rapamycin reactivation during the process of autophagy and contributes to maintenance of muscle homeostasis. PMID:25480788

Front-end electronics development for TPC detector in the MPD/NICA project

NASA Astrophysics Data System (ADS)

Cheremukhina, G.; Movchan, S.; Vereschagin, S.; Zaporozhets, S.

2017-06-01

The article is aimed at describing the development status, measuring results and design changes of the TPC front-end electronics. The TPC is placed in the middle of Multi-Purpose Detector (MPD) and provides tracing and identifying of charged particles in the pseudorapidity range |η| < 1.2. The readout system is one of the most complex parts of the TPC. The electronics of each readout chamber is an independent system. The whole system contains 95232 channels, 1488 64-channel—front-end cards (FEC), 24 readout control units (RCU). The front-end electronics (FEE) is based on ASICs, FPGAs and high-speed serial links. The concept of the TPC front-end electronics has been motivated from one side—by the requirements concerning the NICA accelerator complex which will operate at the luminosity up to 1027 cm-2 s-1 for Au79+ ions over the energy range of 4 < √SNN < 11 GeV with the trigger rate up to 7 kHz and from the other side—by the requirements of the 4-π geometry to minimize the substance on the end-caps of the TPC.

Loss of an Androgen-Inactivating and Isoform-Specific HSD17B4 Splice Form Enables Emergence of Castration-Resistant Prostate Cancer.

PubMed

Ko, Hyun-Kyung; Berk, Michael; Chung, Yoon-Mi; Willard, Belinda; Bareja, Rohan; Rubin, Mark; Sboner, Andrea; Sharifi, Nima

2018-01-16

Castration-resistant prostate cancer (CRPC) requires tumors to engage metabolic mechanisms that allow sustained testosterone and/or dihydrotestosterone to stimulate progression. 17β-Hydroxysteroid dehydrogenase type 4 (17βHSD4), encoded by HSD17B4, is thought to inactivate testosterone and dihydrotestosterone by converting them to their respective inert 17-keto steroids. Counterintuitively, HSD17B4 expression increases in CRPC and predicts poor prognosis. Here, we show that, of five alternative splice forms, only isoform 2 encodes an enzyme capable of testosterone and dihydrotestosterone inactivation. In contrast with other transcripts, functional expression of isoform 2 is specifically suppressed in development of CRPC in patients. Genetically silencing isoform 2 shifts the metabolic balance toward 17β-OH androgens (testosterone and dihydrotestosterone), stimulating androgen receptor (AR) and CRPC development. Our studies specifically implicate HSD17B4 isoform 2 loss in lethal prostate cancer. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Fabrication of Mesoporous Silica/Alumina Hybrid Membrane Film Nanocomposites using Template Sol-Gel Synthesis of Amphiphilic Triphenylene

NASA Astrophysics Data System (ADS)

Lintang, H. O.; Jalani, M. A.; Yuliati, L.; Salleh, M. M.

2017-05-01

Herein we reported that by introducing a one-dimensional (1D) substrate with a porous structure such as anodic aluminum oxide (AAO) membrane, mesoporous silica/alumina hybrid nanocomposites were successfully fabricated by using amphiphilic triphenylene (TPC10TEG) as a template in sol-gel synthesis (TPC10TEG/silicahex). For the optical study of the nanocomposites, TPC10TEG/silicahex showed absorption peak at 264 nm due to the ordered and long-range π-π stacking of the disc-like aromatic triphenylene core. Moreover, the hexagonal arrangement of TPC10TEG/silicahex was proven based on their diffraction peaks of d 100 and d 200 at 2θ = 2.52° and 5.04° and images of transmission electron microscopy (TEM), respectively. For fabrication of mesoporous silica/alumina hybrid membrane, TPC10TEG/silicahex was drop-casted onto AAO membrane for penetration into the porous structure via gravity. X-ray diffraction (XRD) analysis on the resulted hybrid nanocomposites showed that the diffraction peaks of d 100 and d 200 of TPC10TEG/silicahex were still preserved, indicating that the hexagonal arrangements of mesoporous silica were maintained even on AAO substrate. The morphology study on the hybrid nanocomposites using TEM, scanning electron microscope (SEM) and field emission scanning electron microscope (FE-SEM) showed the successful filling of most AAO channels with the TPC10TEG/silicahex nanocomposites.

The influence of ripening stage and cultivation system on the total antioxidant activity and total phenolic compounds of yellow passion fruit pulp.

PubMed

Macoris, Mariana S; De Marchi, Renata; Janzantti, Natália S; Monteiro, Magali

2012-07-01

This work aimed to investigate the influence of both ripening stage and cultivation system on the total phenolic compounds (TPC) and total antioxidant activity (TAA) of passion fruit pulp. TPC extraction was optimized using a 2³ central composed design. The variables were fruit pulp volume, methanol volume and extraction solution volume. TPC was determined using the Folin-Ciocalteu reaction, and TAA using the ABTS radical reaction. The conditions to extract TPC were 2 mL passion fruit pulp and 9 mL extraction solution containing 40% methanol:water (v/v). TPC values increased in the passion fruit pulp during ripening for both cultivation systems, ranging from 281.8 to 361.9 mg gallic acid L⁻¹ (P ≤ 0.05) for the organic pulp and from 291.0 to 338.6 mg gallic acid L⁻¹ (P ≤ 0.05) for the conventional pulp. TPC values increased during ripening for both organic and conventional passion fruit. The same was true for TAA values for conventional passion fruit. For organic passion fruit, however, TAA values were highest at the initial ripening stages. These results suggest that antioxidant compounds exert strong influence on the initial ripening stages for organic passion fruit, when TPC still did not reach its maximum level. Copyright © 2012 Society of Chemical Industry.

Antioxidant and anti-glycation activities correlates with phenolic composition of tropical medicinal herbs.

PubMed

Ramkissoon, J S; Mahomoodally, M F; Ahmed, N; Subratty, A H

2013-07-01

To determine the contribution of total phenolic content (TPC) in glycation inhibitory activity of common tropical medicinal food and spices with potential antioxidative properties. In vitro glucose-bovine serum albumin (BSA) assay was used. Ethanolic extracts of ten common household condiments/herbs (Allium sativum, Zingiber officinale, Thymus vulgaris, Petroselinum crispum, Murraya koenigii Spreng, Mentha piperita L., Curcuma longa L., Allium cepa L., Allium fistulosum and Coriandrum sativum L.) were evaluated for antioxidative activity by 2,2-diphenyl-2-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) and the TPC, flavonoid and tannins content were determined. Findings showed good correlation between TPC/DPPH (r = 0.8), TPC/FRAP (r = 0.8), TPC/anti-glycation (r = 0.9), DPPH/anti-glycation (r = 0.6), FRAP/anti-glycation (r = 0.9), Flavonoid/anti-glycation (r = 0.7) and Tannins/anti-glycation (r = 0.8) and relatively fair correlation for TPC/Flavonoids (r = 0.5) and TPC/Tannins (r = 0.5). Results imply that these plants are potential sources of natural antioxidants which have free radical scavenging activity and might be used for reducing oxidative stress. The positive glycation inhibitory and antioxidative activities of these tropical herbs suggest a possible role in targeting ageing, diabetic complications and oxidative stress related diseases. Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

N Termini of apPDE4 Isoforms Are Responsible for Targeting the Isoforms to Different Cellular Membranes

ERIC Educational Resources Information Center

Jang, Deok-Jin; Park, Soo-Won; Lee, Jin-A; Lee, Changhoon; Chae, Yeon-Su; Park, Hyungju; Kim, Min-Jeong; Choi, Sun-Lim; Lee, Nuribalhae; Kim, Hyoung; Kaang, Bong-Kiun

2010-01-01

Phosphodiesterases (PDEs) are known to play a key role in the compartmentalization of cAMP signaling; however, the molecular mechanisms underlying intracellular localization of different PDE isoforms are not understood. In this study, we have found that each of the supershort, short, and long forms of apPDE4 showed distinct localization in the…

The Capabilities of the upgraded MIPP experiment with respect to Hypernuclear physics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Raja, Rajendran

2012-01-01

We describe the state of analysis of the MIPP experiment, its plans to upgrade the experiment and the impact such an upgraded experiment will have on hypernuclear physics. The upgraded MIPP experiment is designed to measure the properties of strong interaction spectra form beams {pi}{sup {+-}}, K{sup {+-}}, and p{sup {+-}}, for momenta ranging from 1 GeV/c to 120 GeV/c. The layout of the apparatus in the data taken so far can be seen in Figure 1. The centerpiece of the experiment is the time projection chamber, which is followed by the time of flight counter, a multi-cell Cerenkov detectormore » and the RICH detector. The TPC can identify charged particles with momenta less than 1 GeV/c using dE/dx, the time of flight will identify particles below approximately 2 GeV/c, the multi-cell Cerenkov detector is operational from 2.5 GeV/c to 14 GeV/c and the RICH detector can identify particles up to 120 GeVc. Following this is an EM and hadronic calorimeter capable of detecting forward going neutrons and photons. The experiment has been busy analyzing its data taken on various nuclei and beam conditions. The table 2 shows the data taken by MIPP I to date. We have almost complete acceptance in the forward hemisphere in the lab using the TPC. The reconstruction capabilities of the TPC can be seen in Figure 3. The particle identification capabilities of the TPC can be seen in Figure 4. The time of flight system provides further measurement of the particles with momenta less than 2 GeV/c. Figure 5 shows the time of flight data where a kaon peak is clearly visible.« less

Shikonin Inhibites Migration and Invasion of Thyroid Cancer Cells by Downregulating DNMT1

PubMed Central

Zhang, Yue; Sun, Bin; Huang, Zhi

2018-01-01

Background Shikonin is a component of Chinese herbal medicine. The aim of this study was to investigate the effects of shikonin on cell migration of papillary thyroid cancer cells of the TPC-1 cell line in vitro and expression levels of the phosphate and tensin homolog deleted on chromosome 10 (PTEN) and DNA methyltransferase 1 (DNMT1) genes. Material/Methods The Cell Counting Kit-8 (CCK-8) assay was performed to evaluate the proliferation of TPC-1 papillary thyroid cancer cells, and the normal thyroid cells, HTori-3, in vitro. A transwell motility assay was used to analyze the migration of TPC-1 cells. Western blot was performed to determine the expression levels of PTEN and DNMT1 genes. A methylation-specific polymerase chain reaction (PCR) (MSP) assay was used to evaluate the methylation of PTEN. Results Following treatment with shikonin, the cell survival rate of TPC-1 cells decreased in a dose-dependent manner; the inhibitory effects on HTori-3 cells were less marked. Shikonin inhibited TPC-1 cell migration and invasion in a dose-dependent manner. The methylation of PTEN was suppressed by shikonin, which also reduced the expression of DNMT1 in a dose-dependent manner, and increased the expression of PTEN. Overexpression of DNMT1 promoted the migration of TPC-1 cells and the methylation of PTEN. Levels of protein expression of PTEN in TPC-1 cells treated with shikonin decreased, and were increased by DNMT1 knockdown. Conclusions Shikonin suppressed the expression of DNMT1, reduced PTEN gene methylation, and increased PTEN protein expression, leading to the inhibition of TPC-1 cell migration. PMID:29389913

A streamlined Python framework for AT-TPC data analysis

NASA Astrophysics Data System (ADS)

Taylor, J. Z.; Bradt, J.; Bazin, D.; Kuchera, M. P.

2017-09-01

User-friendly data analysis software has been developed for the Active-Target Time Projection Chamber (AT-TPC) experiment at the National Superconducting Cyclotron Laboratory at Michigan State University. The AT-TPC, commissioned in 2014, is a gas-filled detector that acts as both the detector and target for high-efficiency detection of low-intensity, exotic nuclear reactions. The pytpc framework is a Python package for analyzing AT-TPC data. The package was developed for the analysis of 46Ar(p, p) data. The existing software was used to analyze data produced by the 40Ar(p, p) experiment that ran in August, 2015. Usage of the package was documented in an analysis manual both to improve analysis steps and aid in the work of future AT-TPC users. Software features and analysis methods in the pytpc framework will be presented along with the 40Ar results.

TPC status for MPD experiment of NICA project

NASA Astrophysics Data System (ADS)

Averyanov, A.; Bazhazhin, A.; Chepurnov, V. F.; Chepurnov, V. V.; Cheremukhina, G.; Chernenko, S.; Fateev, O.; Kiriushin, Yu.; Kolesnikov, A.; Korotkova, A.; Levchanovsky, F.; Lukstins, J.; Movchan, S.; Pilyar, A.; Razin, S.; Ribakov, A.; Samsonov, V.; Vereschagin, S.; Zanevsky, Yu.; Zaporozhets, S.; Zruev, V.

2017-06-01

In a frame of the JINR scientific program on study of hot and dense baryonic matter a new accelerator complex Ion Collider fAcility (NICA) based on the Nuclotron-M is under realization. It will operate at luminosity up to 1027 cm-2s-1 for Au79+ ions. Two interaction points are foreseen at NICA for two detectors which will operate simultaneously. One of these detectors, the Multi-Purpose Detector (MPD), is optimized for investigations of heavy-ion collisions. The Time-Projection Chamber (TPC) is the main tracking detector of the MPD central barrel. It is a well-known detector for 3-dimensional tracking and particle identification for high multiplicity events. The conceptual layout of MPD and detailed description of the design and main working parameters of TPC, the readout system based on MWPC and readout electronics as well as the TPC subsystems and tooling for assembling and integration TPC into MPD are presented.

Visualizing Time Projection Chamber Data for Education and Outreach

NASA Astrophysics Data System (ADS)

Crosby, Jacob

2017-09-01

The widespread availability of portable computers in the form of smartphones provides a unique opportunity to introduce scientific concepts to a broad audience, for the purpose of education, or for the purpose of sharing exciting developments and research. Unity, a free game development platform, has been used to develop a program to visualize 3-D events from a Time Projection Chamber (TPC). The program can be presented as a Virtual Reality (VR) application on a smartphone, which can serve as a standalone demonstration for interested individuals, or as a resource for educators. An interactive experience to watch nuclear events unfold demonstrates the principles of particle detection with a TPC, as well as providing information about the particles present. Different kinds of reactions can be showcased. The current state of tools within this program for outreach and educational purposes will be highlighted and presented in this poster, along with key design concerns and optimizations necessary for running an interactive VR app. The events highlighted in this program are from the S πRIT TPC, but the program can be applied to other 3-D detectors. This work is supported by the U.S. Department of Energy under Grant Nos. DE-SC0014530, DE-NA0002923 and US NSF under Grant No. PHY-1565546.

A Venom Serpin Splicing Isoform of the Endoparasitoid Wasp Pteromalus puparum Suppresses Host Prophenoloxidase Cascade by Forming Complexes with Host Hemolymph Proteinases*

PubMed Central

Yan, Zhichao; Fang, Qi; Liu, Yang; Xiao, Shan; Yang, Lei; Wang, Fei; An, Chunju; Werren, John H.; Ye, Gongyin

2017-01-01

To ensure successful parasitism, parasitoid wasps inject venom along with their eggs into their hosts. The venom serves to suppress host immune responses, including melanization. Venom from Pteromalus puparum, a pupal endoparasitoid, inhibits melanization of host hemolymph in vitro in a dose-dependent manner. Using assay-guided fractionation, a serpin splicing isoform with phenoloxidase inhibitory activity was identified as P. puparum serpin-1, venom isoform (PpS1V). This serpin gene has 16 predicted splicing isoforms that differ only in the C-terminal region. RT-PCR results show that the specific serpin isoform is differentially expressed in the venom gland. Recombinant PpS1V (rPpS1V) suppresses host prophenoloxidase (PPO) activation rather than inhibiting the phenoloxidase directly. Pulldown assays show that PpS1V forms complexes with two host hemolymph proteins, here named Pieris rapae hemolymph proteinase 8 (PrHP8) and P. rapae prophenoloxidase-activating proteinase 1 (PrPAP1), based on gene sequence blasting and phylogenetic analysis. The role of rPrPAP1 in the PPO activation cascade and its interaction with rPpS1V were confirmed. The stoichiometry of inhibition of PrPAP1 by PpS1V is 2.3. PpS1V also inhibits PPO activation in a non-natural host, Ostrinia furnacalis, through forming a complex with O. furnacalis serine protease 13 (OfSP13), an ortholog to PrPAP1. Our results identify a venom-enriched serpin isoform in P. puparum that inhibits host PPO activation, probably by forming a complex with host hemolymph proteinase PrPAP1. PMID:27913622

Selective Suppression of Endothelial Cell Apoptosis by the High Molecular Weight Form of Adiponectin

PubMed Central

Kobayashi, Hideki; Ouchi, Noriyuki; Kihara, Shinji; Walsh, Kenneth; Kumada, Masahiro; Abe, Yuki; Funahashi, Tohru; Matsuzawa, Yuji

2015-01-01

Adiponectin is an adipocyte-derived, antiatherogenic protein that is present in serum as three isoforms. Total adiponectin levels are decreased in obese or diabetic humans or animal models. This study was designed to elucidate the relative isoform distribution of adiponectin in human disease states and identify the active form of adiponectin toward vascular endothelial cells. The percentage of high molecular weight form (HMW) per total adiponectin was significantly lower in patients with coronary artery disease than control subjects, whereas the hexamer form was similar and the trimer form was significantly higher. During weight reduction in obese subjects, the HMW form increased and the trimer and hexamer forms decreased. Recombinant adiponectin dose-dependently suppressed apoptosis and caspase-3 activity in human umbilical vein endothelial cells (HUVECs). Transduction with dominant-negative AMP-activated protein kinase (AMPK) abolished the suppressive effect of adiponectin on HUVECs. Gel filtration chromatography was used to separate the adiponectin isoforms, and the antiapoptotic effect toward HUVECs was only observed with the HMW form. These data suggest that HMW adiponectin specifically confers the vascular-protective activities of this adipocytokine. PMID:14752031

Enhanced trigger for the NIFFTE fissionTPC in presence of high-rate alpha backgrounds

NASA Astrophysics Data System (ADS)

Bundgaard, Jeremy; Niffte Collaboration

2015-10-01

Nuclear physics and nuclear energy communities call for new, high precision measurements to improve existing fission models and design next generation reactors. The Neutron Induced Fission Fragment Tracking experiment (NIFFTE) has developed the fission Time Projection Chamber (fissionTPC) to measure neutron induced fission with unrivaled precision. The fissionTPC is annually deployed to the Weapons Neutron Research facility at Los Alamos Neutron Science Center where it operates with a neutron beam passing axially through the drift volume, irradiating heavy actinide targets to induce fission. The fissionTPC was developed at the Lawrence Livermore National Laboratory's TPC lab, where it measures spontaneous fission from radioactive sources to characterize detector response, improve performance, and evolve the design. To measure 244Cm, we've developed a fission trigger to reduce the data rate from alpha tracks while maintaining a high fission detection efficiency. In beam, alphas from 239Pu are a large background when detecting fission fragments; implementing the fission trigger will greatly reduce this background. The implementation of the cathode fission trigger in the fissionTPC will be presented along with a detailed study of its efficiency.

Noise Characterization and Filtering in the MicroBooNE Liquid Argon TPC

NASA Astrophysics Data System (ADS)

Acciarri, R.; Adams, C.; An, R.; Anthony, J.; Asaadi, J.; Auger, M.; Bagby, L.; Balasubramanian, S.; Baller, B.; Barnes, C.; Barr, G.; Bass, M.; Bay, F.; Bishai, M.; Blake, A.; Bolton, T.; Bullard, B.; Camilleri, L.; Caratelli, D.; Carls, B.; Castillo Fernandez, R.; Cavanna, F.; Chen, H.; Church, E.; Cianci, D.; Cohen, E.; Collin, G. H.; Conrad, J. M.; Convery, M.; Crespo-Anadón, J. I.; De Geronimo, G.; Del Tutto, M.; Devitt, D.; Dytman, S.; Eberly, B.; Ereditato, A.; Escudero Sanchez, L.; Esquivel, J.; Fadeeva, A. A.; Fleming, B. T.; Foreman, W.; Furmanski, A. P.; Garcia-Gamez, D.; Garvey, G. T.; Genty, V.; Goeldi, D.; Gollapinni, S.; Graf, N.; Gramellini, E.; Greenlee, H.; Grosso, R.; Guenette, R.; Hackenburg, A.; Hamilton, P.; Hen, O.; Hewes, J.; Hill, C.; Ho, J.; Horton-Smith, G.; Hourlier, A.; Huang, E.-C.; James, C.; de Vries, J. Jan; Jen, C.-M.; Jiang, L.; Johnson, R. A.; Joshi, J.; Jostlein, H.; Kaleko, D.; Karagiorgi, G.; Ketchum, W.; Kirby, B.; Kirby, M.; Kobilarcik, T.; Kreslo, I.; Laube, A.; Li, S.; Li, Y.; Lister, A.; Littlejohn, B. R.; Lockwitz, S.; Lorca, D.; Louis, W. C.; Luethi, M.; Lundberg, B.; Luo, X.; Marchionni, A.; Mariani, C.; Marshall, J.; Martinez Caicedo, D. A.; Meddage, V.; Miceli, T.; Mills, G. B.; Moon, J.; Mooney, M.; Moore, C. D.; Mousseau, J.; Murrells, R.; Naples, D.; Nienaber, P.; Nowak, J.; Palamara, O.; Paolone, V.; Papavassiliou, V.; Pate, S. F.; Pavlovic, Z.; Piasetzky, E.; Porzio, D.; Pulliam, G.; Qian, X.; Raaf, J. L.; Radeka, V.; Rafique, A.; Rescia, S.; Rochester, L.; von Rohr, C. Rudolf; Russell, B.; Schmitz, D. W.; Schukraft, A.; Seligman, W.; Shaevitz, M. H.; Sinclair, J.; Smith, A.; Snider, E. L.; Soderberg, M.; Söldner-Rembold, S.; Soleti, S. R.; Spentzouris, P.; Spitz, J.; St. John, J.; Strauss, T.; Szelc, A. M.; Tagg, N.; Terao, K.; Thomson, M.; Thorn, C.; Toups, M.; Tsai, Y.-T.; Tufanli, S.; Usher, T.; Van De Pontseele, W.; Van de Water, R. G.; Viren, B.; Weber, M.; Wickremasinghe, D. A.; Wolbers, S.; Wongjirad, T.; Woodruff, K.; Yang, T.; Yates, L.; Yu, B.; Zeller, G. P.; Zennamo, J.; Zhang, C.

2017-08-01

The low-noise operation of readout electronics in a liquid argon time projection chamber (LArTPC) is critical to properly extract the distribution of ionization charge deposited on the wire planes of the TPC, especially for the induction planes. This paper describes the characteristics and mitigation of the observed noise in the MicroBooNE detector. The MicroBooNE's single-phase LArTPC comprises two induction planes and one collection sense wire plane with a total of 8256 wires. Current induced on each TPC wire is amplified and shaped by custom low-power, low-noise ASICs immersed in the liquid argon. The digitization of the signal waveform occurs outside the cryostat. Using data from the first year of MicroBooNE operations, several excess noise sources in the TPC were identified and mitigated. The residual equivalent noise charge (ENC) after noise filtering varies with wire length and is found to be below 400 electrons for the longest wires (4.7 m). The response is consistent with the cold electronics design expectations and is found to be stable with time and uniform over the functioning channels. This noise level is significantly lower than previous experiments utilizing warm front-end electronics.

Subcellular distribution of serine acetyltransferase from Pisum sativum and characterization of an Arabidopsis thaliana putative cytosolic isoform.

PubMed

Ruffet, M L; Lebrun, M; Droux, M; Douce, R

1995-01-15

The intracellular compartmentation of serine acetyltransferase, a key enzyme in the L-cysteine biosynthesis pathway, has been investigated in pea (Pisum sativum) leaves, by isolation of organelles and fractionation of protoplasts. Enzyme activity was mainly located in mitochondria (approximately 76% of total cellular activity). Significant activity was also identified in both the cytosol (14% of total activity) and chloroplasts (10% of total activity). Three enzyme forms were separated by anion-exchange chromatography, and each form was found to be specific for a given intracellular compartment. To obtain cDNA encoding the isoforms, functional complementation experiments were performed using an Arabidopsis thaliana expression library and an Escherichia coli mutant devoid of serine acetyltransferase activity. This strategy allowed isolation of three distinct cDNAs encoding serine acetyltransferase isoforms, as confirmed by enzyme activity measurements, genomic hybridizations, and nucleotide sequencing. The cDNA and related gene for one of the three isoforms have been characterized. The predicted amino acid sequence shows that it encodes a polypeptide of M(r) 34,330 exhibiting 41% amino acid identity with the E. coli serine acetyltransferase. Since none of the general features of transit peptides could be observed in the N-terminal region of this isoform, we assume that it is a cytosolic form.

Role of acyl carrier protein isoforms in plant lipid metabolism: Progress report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ohlrogge, J.B.

1989-01-01

Previous research from my lab has revealed that several higher plant species have multiple isoforms of acyl carrier protein (ACP) and therefore this trait appears highly conserved among higher plants. This level of conservation suggests that the existence of ACP isoforms is not merely the results of neutral gene duplications. We have developed techniques to examine a wider range of species. Acyl carrier proteins can be labelled very specifically and to high specific activity using H-palmitate and the E. coli enzyme acyl-ACP synthetase. Isoforms were then resolved by western blotting and native PAGE of H-palmitate labelled ACP's. Multiple isoforms ofmore » ACP were observed the leaf tissue of the monocots Avena sativa and Hordeum vulgare and dicots including Arabidopsis thallina, Cuphea wrightii, and Brassica napus. Lower vascular plants including the cycad, Dioon edule, Ginkgo biloba, the gymnosperm Pinus, the fern Anernia phyllitidis and Psilotum nudum, the most primitive known extant vascular plant, were also found to have multiple ACP isoforms as were the nonvascular liverwort, Marchantia and moss, Polytrichum. Therefore, the development of ACP isoforms occurred early in evolution. However, the uniellular alge Chlamydomonas and Dunaliella and the photosynthetic cyanobacteria Synechocystis and Agmnellum have only a single elecrophotetic form of ACP. Thus, multiple forms of ACP do not occur in all photosynthetic organisms but may be associated with multicellular plants.« less

Purification and Characterization of Two Voltage-Dependent Anion Channel Isoforms from Plant Seeds1

PubMed Central

Abrecht, Helge; Wattiez, Ruddy; Ruysschaert, Jean-Marie; Homblé, Fabrice

2000-01-01

Mitochondria were isolated from imbibed seeds of lentil (Lens culinaris) and Phaseolus vulgaris. We copurified two voltage-dependent anion channel from detergent solubilized mitochondria in a single purification step using hydroxyapatite. The two isoforms from P. vulgaris were separated by chromatofocusing chromatography in 4 m urea without any loss of channel activity. Channel activity of each isoform was characterized upon reconstitution into diphytanoyl phosphatidylcholine planar lipid bilayers. Both isoforms form large conductance channels that are slightly anion selective and display cation selective substates. PMID:11080295

The human two-pore channel 1 is modulated by cytosolic and luminal calcium

PubMed Central

Lagostena, Laura; Festa, Margherita; Pusch, Michael; Carpaneto, Armando

2017-01-01

Two-pore channels (TPC) are intracellular endo-lysosomal proteins with only recently emerging roles in organellar signalling and involvement in severe human diseases. Here, we investigated the functional properties of human TPC1 expressed in TPC-free vacuoles from Arabidopsis thaliana cells. Large (20 pA/pF) TPC1 currents were elicited by cytosolic addition of the phosphoinositide phosphatidylinositol-(3,5)-bisphosphate (PI(3,5)P2) with an apparent binding constant of ~15 nM. The channel is voltage-dependent, activating at positive potentials with single exponential kinetics and currents are Na+ selective, with measurable but low permeability to Ca2+. Cytosolic Ca2+ modulated hTPC1 in dual way: low μM cytosolic Ca2+ increased activity by shifting the open probability towards negative voltages and by accelerating the time course of activation. This mechanism was well-described by an allosteric model. Higher levels of cytosolic Ca2+ induced a voltage-dependent decrease of the currents compatible with Ca2+ binding in the permeation pore. Conversely, an increase in luminal Ca2+ decreased hTPC1 activity. Our data point to a process in which Ca2+ permeation in hTPC1 has a positive feedback on channel activity while Na+ acts as a negative regulator. We speculate that the peculiar Ca2+ and Na+ dependence are key for the physiological roles of the channel in organellar homeostasis and signalling. PMID:28252105

Adaptive molecular evolution of the two-pore channel 1 gene TPC1 in the karst-adapted genus Primulina (Gesneriaceae)

PubMed Central

Tao, Junjie; Feng, Chao; Ai, Bin; Kang, Ming

2016-01-01

Background and Aims Limestone karst areas possess high floral diversity and endemism. The genus Primulina, which contributes to the unique calcicole flora, has high species richness and exhibit specific soil-based habitat associations that are mainly distributed on calcareous karst soils. The adaptive molecular evolutionary mechanism of the genus to karst calcium-rich environments is still not well understood. The Ca2+-permeable channel TPC1 was used in this study to test whether its gene is involved in the local adaptation of Primulina to karst high-calcium soil environments. Methods Specific amplification and sequencing primers were designed and used to amplify the full-length coding sequences of TPC1 from cDNA of 76 Primulina species. The sequence alignment without recombination and the corresponding reconstructed phylogeny tree were used in molecular evolutionary analyses at the nucleic acid level and amino acid level, respectively. Finally, the identified sites under positive selection were labelled on the predicted secondary structure of TPC1. Key Results Seventy-six full-length coding sequences of Primulina TPC1 were obtained. The length of the sequences varied between 2220 and 2286 bp and the insertion/deletion was located at the 5′ end of the sequences. No signal of substitution saturation was detected in the sequences, while significant recombination breakpoints were detected. The molecular evolutionary analyses showed that TPC1 was dominated by purifying selection and the selective pressures were not significantly different among species lineages. However, significant signals of positive selection were detected at both TPC1 codon level and amino acid level, and five sites under positive selective pressure were identified by at least three different methods. Conclusions The Ca2+-permeable channel TPC1 may be involved in the local adaptation of Primulina to karst Ca2+-rich environments. Different species lineages suffered similar selective pressure associated with calcium in karst environments, and episodic diversifying selection at a few sites may play a major role in the molecular evolution of Primulina TPC1. PMID:27582362

Impregnation quality of shredded semipreg after compression moulding

NASA Astrophysics Data System (ADS)

Vincent, G.; Balakrishnan, V.; de Bruijn, T. A.; Wijskamp, S.; Abdul Rasheed, M. I.

2017-10-01

Manufacturing of thermoplastic composites (TPC) inherently generates scrap, mainly in the form of offcuts or rejected parts. The growth of TPC over recent decades has now reached a point where developing specific recycling solutions for TPC waste has become crucial. While looking at the various steps during which scrap is produced, the nesting of semipreg or prepreg appears to be critical. This work aims to develop a route for recycling semipreg offcuts, comprising shredding and compression moulding. This article focuses on an experimental study of the compression moulding step of carbon fibres reinforced PPS (C/PPS) to investigate the uniformity and impregnation quality of plates. These plates were realised in a picture frame while varying both the fibre volume content between 30% and 50% and the processing parameters. Visual inspection and cross-sectional microscopy were performed to assess the quality of each plate. As a first step, the influence of the type of added matrix (film, powder, pellets) and the type of pre-impregnation (film, powder) was studied. Stacking of polymer powder with shredded powder-coated semipreg gave the best impregnation quality. It was also shown that longer dwell time at melt leads to better consolidation quality. However, the difficulty in obtaining good impregnation comes from the disentangled shredded material, which is composed of three forms: semipreg flakes, dry bundles and pieces of matrix. When dry bundles reach the mould surfaces during the filling of the mould or when they are packed together, the consolidation cycle hardly impregnates them and they remain dry afterwards. Furthermore, large local variations of fibre fraction were noticed, resulting from a random mould filling. Therefore, the recycling solution for shredded semipreg is feasible when the fibre fraction is reduced but improvements on the part variability still have to be made.

Comparative Study of Antioxidant Power, Polyphenols, Flavonoids and Betacyanins of the Peel and Pulp of Three Tunisian Opuntia Forms

PubMed Central

Yeddes, Nizar; Chérif, Jamila K.; Guyot, Sylvain; Sotin, Hélène; Ayadi, Malika T.

2013-01-01

The antioxidant activity and the chemical composition of methanol extracts from peel and pulp belonging to two species of Tunisian prickly pears Opuntia ficus indica (spiny and thornless forms) and Opuntia stricta have been studied. The antioxidant capacity was measured by DPPH radical scavenging activity. The total phenolic compound (TPC) and the total flavonoid content were determined by the Folin–Ciocalteu method and colorimetric method, respectively. The phenolic compounds were identified and quantified by high-performance liquid chromatography (HPLC) coupled with an electrospray ionization mass spectrometry (ESI-MS). The results showed that O. stricta fruits present the best antioxidant activities than the two forms of O. ficus indica, while the TPC was more important in O. ficus indica than in the O. stricta fruits. The peels have higher flavonoids than pulp, and the thornless variety has more flavonoid than the spiny. The RP-HPLC and ESI-MS analysis detected two classes of phenolic compounds and betalain pigments. Isorhamnetin derivatives are the dominant flavonol glycoside identified in O. ficus indica (spiny: 65.25 μg·g−1; thornless: 77.03 μg·g−1) and O. stricta peels (19.22 μg·g−1). PMID:26787622

High susceptibility to fatty liver disease in two-pore channel 2-deficient mice.

PubMed

Grimm, Christian; Holdt, Lesca M; Chen, Cheng-Chang; Hassan, Sami; Müller, Christoph; Jörs, Simone; Cuny, Hartmut; Kissing, Sandra; Schröder, Bernd; Butz, Elisabeth; Northoff, Bernd; Castonguay, Jan; Luber, Christian A; Moser, Markus; Spahn, Saskia; Lüllmann-Rauch, Renate; Fendel, Christina; Klugbauer, Norbert; Griesbeck, Oliver; Haas, Albert; Mann, Matthias; Bracher, Franz; Teupser, Daniel; Saftig, Paul; Biel, Martin; Wahl-Schott, Christian

2014-08-21

Endolysosomal organelles play a key role in trafficking, breakdown and receptor-mediated recycling of different macromolecules such as low-density lipoprotein (LDL)-cholesterol, epithelial growth factor (EGF) or transferrin. Here we examine the role of two-pore channel (TPC) 2, an endolysosomal cation channel, in these processes. Embryonic mouse fibroblasts and hepatocytes lacking TPC2 display a profound impairment of LDL-cholesterol and EGF/EGF-receptor trafficking. Mechanistically, both defects can be attributed to a dysfunction of the endolysosomal degradation pathway most likely on the level of late endosome to lysosome fusion. Importantly, endolysosomal acidification or lysosomal enzyme function are normal in TPC2-deficient cells. TPC2-deficient mice are highly susceptible to hepatic cholesterol overload and liver damage consistent with non-alcoholic fatty liver hepatitis. These findings indicate reduced metabolic reserve of hepatic cholesterol handling. Our results suggest that TPC2 plays a crucial role in trafficking in the endolysosomal degradation pathway and, thus, is potentially involved in the homoeostatic control of many macromolecules and cell metabolites.

Performance study of the neutron-TPC

NASA Astrophysics Data System (ADS)

Huang, Meng; Li, Yulan; Niu, Libo; Deng, Zhi; Cheng, Xiaolei; He, Li; Zhang, Hongyan; Fu, Jianqiang; Yan, Yangyang; Cai, Yiming; Li, Yuanjing

2017-02-01

Fast neutron spectrometers will play an important role in the future of the nuclear industry and nuclear physics experiments, in tasks such as fast neutron reactor monitoring, thermo-nuclear fusion plasma diagnostics, nuclear reaction cross-section measurement, and special nuclear material detection. Recently, a new fast neutron spectrometer based on a GEM (Gas Electron Multiplier amplification)-TPC (Time Projection Chamber), named the neutron-TPC, has been under development at Tsinghua University. It is designed to have a high energy resolution, high detection efficiency, easy access to the medium material, an outstanding n/γ suppression ratio, and a wide range of applications. This paper presents the design, test, and experimental study of the neutron-TPC. Based on the experimental results, the energy resolution (FWHM) of the neutron-TPC can reach 15.7%, 10.3% and 7.0% with detection efficiency higher than 10-5 for 1.2 MeV, 1.81 MeV and 2.5 MeV neutrons respectively. Supported by National Natural Science Foundation of China (11275109)

Noise Characterization and Filtering in the MicroBooNE Liquid Argon TPC

DOE Office of Scientific and Technical Information (OSTI.GOV)

Acciarri, R.; et al.

The low-noise operation of readout electronics in a liquid argon time projection chamber (LArTPC) is critical to properly extract the distribution of ionization charge deposited on the wire planes of the TPC, especially for the induction planes. This paper describes the characteristics and mitigation of the observed noise in the MicroBooNE detector. The MicroBooNE's single-phase LArTPC comprises two induction planes and one collection sense wire plane with a total of 8256 wires. Current induced on each TPC wire is amplified and shaped by custom low-power, low-noise ASICs immersed in the liquid argon. The digitization of the signal waveform occurs outsidemore » the cryostat. Using data from the first year of MicroBooNE operations, several excess noise sources in the TPC were identified and mitigated. The residual equivalent noise charge (ENC) after noise filtering varies with wire length and is found to be below 400 electrons for the longest wires (4.7 m). The response is consistent with the cold electronics design expectations and is found to be stable with time and uniform over the functioning channels. This noise level is significantly lower than previous experiments utilizing warm front-end electronics.« less

Noise Characterization and Filtering in the MicroBooNE Liquid Argon TPC

DOE Office of Scientific and Technical Information (OSTI.GOV)

Acciarri, R.; Adams, C.; An, R.

The low-noise operation of readout electronics in a liquid argon time projection chamber (LArTPC) is critical to properly extract the distribution of ionization charge deposited on the wire planes of the TPC, especially for the induction planes. This paper describes the characteristics and mitigation of the observed noise in the MicroBooNE detector. The MicroBooNE's single-phase LArTPC comprises two induction planes and one collection sense wire plane with a total of 8256 wires. Current induced on each TPC wire is amplified and shaped by custom low-power, low-noise ASICs immersed in the liquid argon. The digitization of the signal waveform occurs outsidemore » the cryostat. Using data from the first year of MicroBooNE operations, several excess noise sources in the TPC were identified and mitigated. The residual equivalent noise charge (ENC) after noise filtering varies with wire length and is found to be below 400 electrons for the longest wires (4.7 m). The response is consistent with the cold electronics design expectations and is found to be stable with time and uniform over the functioning channels. In conclusion, this noise level is significantly lower than previous experiments utilizing warm front-end electronics.« less

Naringenin Impairs Two-Pore Channel 2 Activity And Inhibits VEGF-Induced Angiogenesis.

PubMed

Pafumi, Irene; Festa, Margherita; Papacci, Francesca; Lagostena, Laura; Giunta, Cristina; Gutla, Vijay; Cornara, Laura; Favia, Annarita; Palombi, Fioretta; Gambale, Franco; Filippini, Antonio; Carpaneto, Armando

2017-07-11

Our research introduces the natural flavonoid naringenin as a novel inhibitor of an emerging class of intracellular channels, Two-Pore Channel 2 (TPC2), as shown by electrophysiological evidence in a heterologous system, i.e. Arabidopsis vacuoles lacking endogenous TPCs. In view of the control exerted by TPC2 on intracellular calcium signaling, we demonstrated that naringenin dampens intracellular calcium responses of human endothelial cells stimulated with VEGF, histamine or NAADP-AM, but not with ATP or Angiopoietin-1 (negative controls). The ability of naringenin to impair TPC2-dependent biological activities was further explored in an established in vivo model, in which VEGF-containing matrigel plugs implanted in mice failed to be vascularized in the presence of naringenin. Overall, the present data suggest that naringenin inhibition of TPC2 activity and the observed inhibition of angiogenic response to VEGF are linked by impaired intracellular calcium signaling. TPC2 inhibition is emerging as a key therapeutic step in a range of important pathological conditions including the progression and metastatic potential of melanoma, Parkinson's disease, and Ebola virus infection. The identification of naringenin as an inhibitor of TPC2-mediated signaling provides a novel and potentially relevant tool for the advancement of this field of research.

Noise Characterization and Filtering in the MicroBooNE Liquid Argon TPC

DOE PAGES

Acciarri, R.; Adams, C.; An, R.; ...

2017-08-04

The low-noise operation of readout electronics in a liquid argon time projection chamber (LArTPC) is critical to properly extract the distribution of ionization charge deposited on the wire planes of the TPC, especially for the induction planes. This paper describes the characteristics and mitigation of the observed noise in the MicroBooNE detector. The MicroBooNE's single-phase LArTPC comprises two induction planes and one collection sense wire plane with a total of 8256 wires. Current induced on each TPC wire is amplified and shaped by custom low-power, low-noise ASICs immersed in the liquid argon. The digitization of the signal waveform occurs outsidemore » the cryostat. Using data from the first year of MicroBooNE operations, several excess noise sources in the TPC were identified and mitigated. The residual equivalent noise charge (ENC) after noise filtering varies with wire length and is found to be below 400 electrons for the longest wires (4.7 m). The response is consistent with the cold electronics design expectations and is found to be stable with time and uniform over the functioning channels. In conclusion, this noise level is significantly lower than previous experiments utilizing warm front-end electronics.« less

Gating of the two-pore cation channel AtTPC1 in the plant vacuole is based on a single voltage-sensing domain.

PubMed

Jaślan, D; Mueller, T D; Becker, D; Schultz, J; Cuin, T A; Marten, I; Dreyer, I; Schönknecht, G; Hedrich, R

2016-09-01

The two-pore cation channel TPC1 operates as a dimeric channel in animal and plant endomembranes. Each subunit consists of two homologous Shaker-like halves, with 12 transmembrane domains in total (S1-S6, S7-S12). In plants, TPC1 channels reside in the vacuolar membrane, and upon voltage stimulation, give rise to the well-known slow-activating SV currents. Here, we combined bioinformatics, structure modelling, site-directed mutagenesis, and in planta patch clamp studies to elucidate the molecular mechanisms of voltage-dependent channel gating in TPC1 in its native plant background. Structure-function analysis of the Arabidopsis TPC1 channel in planta confirmed that helix S10 operates as the major voltage-sensing site, with Glu450 and Glu478 identified as possible ion-pair partners for voltage-sensing Arg537. The contribution of helix S4 to voltage sensing was found to be negligible. Several conserved negative residues on the luminal site contribute to calcium binding, stabilizing the closed channel. During evolution of plant TPC1s from two separate Shaker-like domains, the voltage-sensing function in the N-terminal Shaker-unit (S1-S4) vanished. © 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

Production of raw-starch-digesting α-amylase isoform from Bacillus sp. under solid-state fermentation and biochemical characterization.

PubMed

Božić, Nataša; Slavić, Marinela Šokarda; Gavrilović, Anja; Vujčić, Zoran

2014-07-01

α-Amylase production by solid-state fermentation of different Bacillus sp. was studied previously on different fermentation media. However, no study has been reported on the influence of selected media on expression of desired amylase isoforms such as raw-starch-digesting amylase (RSDA). In this paper, the influence of different inexpensive and available agro-resources as solid media (corn, wheat and triticale) on α-amylase isoform induction from three wild-type Bacillus sp., selected among one hundred strains tested, namely 9B, 12B and 24A was investigated. For all three strains, tested amylases were detected in the multiple forms; however, number and intensity of each form differed depending on the solid media used for growth. To determine which isoform from Bacillus sp. 12B was RSDA, the suspected isoform was purified. The optimum pH for the purified α-amylase isoform was 6.0-8.0, while the optimum temperature was 60-90 °C. Isoform was considerably thermostable and Ca(2+)-independent, and actually the only α-amylase active towards raw starch. Purification and characterization of RSDA showed that not all of the solid media tested induced RSDA. From an economic point of view, it might be significant to obtain pure isoenzyme for potential use in the raw-starch hydrolysis, since it was 5 times more efficient in raw corn starch hydrolysis than the crude amylase preparation.

Identification of a Novel Splice Variant Isoform of TREM-1 in Human Neutrophil Granules.

PubMed

Baruah, Sankar; Keck, Kathy; Vrenios, Michelle; Pope, Marshall R; Pearl, Merideth; Doerschug, Kevin; Klesney-Tait, Julia

2015-12-15

Triggering receptor expressed on myeloid cells-1 (TREM-1) is critical for inflammatory signal amplification. Humans have two forms of TREM-1: a membrane receptor, associated with the adaptor DAP12, and a soluble receptor detected at times of infection. The membrane receptor isoform acts synergistically with the TLR pathway to promote cytokine secretion and neutrophil migration, whereas the soluble receptor functions as a counterregulatory molecule. In multiple models of sepsis, exogenous administration of soluble forms of TREM-1 attenuates inflammation and markedly improves survival. Despite intense interest in soluble TREM-1, both as a clinical predictor of survival and as a therapeutic tool, the origin of native soluble TREM-1 remains controversial. Using human neutrophils, we identified a 15-kDa TREM-1 isoform in primary (azurophilic) and secondary (specific) granules. Mass spectrometric analysis, ELISA, and immunoblot confirm that the 15-kDa protein is a novel splice variant form of TREM-1 (TREM-1sv). Neutrophil stimulation with Pseudomonas aeruginosa, LPS, or PAM(3)Cys4 resulted in degranulation and release of TREM-1sv. The addition of exogenous TREM-1sv inhibited TREM-1 receptor-mediated proinflammatory cytokine production. Thus, these data reveal that TREM-1 isoforms simultaneously activate and inhibit inflammation via the canonical membrane TREM-1 molecule and this newly discovered granular isoform, TREM-1sv. Copyright © 2015 by The American Association of Immunologists, Inc.

ISOFORMS OF VITAMIN E DIFFERENTIALLY REGULATE INFLAMMATION

PubMed Central

Cook-Mills, Joan M.; McCary, Christine A.

2011-01-01

Vitamin E regulation of disease has been extensively studied in humans, animal models and cell systems. Most of these studies focus on the α-tocopherol isoform of vitamin E. These reports indicate contradictory outcomes for anti-inflammatory functions of the α-tocopherol isoform of vitamin E, especially with regards to clinical studies of asthma and atherosclerosis. These seemingly disparate clinical results are consistent with recently reported unrecognized properties of isoforms of vitamin E. Recently, it has been reported that physiological levels of purified natural forms of vitamin E have opposing regulatory functions during inflammation. These opposing regulatory functions by physiological levels of vitamin E isoforms impact interpretations of previous studies on vitamin E. Moreover, additional recent studies also indicate that the effects of vitamin E isoforms on inflammation are only partially reversible using physiological levels of a vitamin E isoform with opposing immunoregulatory function. Thus, this further influences interpretations of previous studies with vitamin E in which there was inflammation and substantial vitamin E isoforms present before the initiation of the study. In summary, this review will discuss regulation of inflammation by vitamin E, including alternative interpretations of previous studies in the literature with regards to vitamin E isoforms. PMID:20923401

Sealed operation, and circulation and purification of gas in the HARPO TPC

NASA Astrophysics Data System (ADS)

Frotin, M.; Gros, P.; Attié, D.; Bernard, D.; Dauvois, V.; Delbart, A.; Durand, D.; Geerebaert, Y.; Legand, S.; Magnier, P.; Poilleux, P.; Semeniouk, I.

2018-02-01

HARPO is a time projection chamber (TPC) demonstrator of a gamma-ray telescope and polarimeter in the MeV-GeV range, for a future space mission. We present the evolution of the TPC performance over a five month sealed-mode operation, by the analysis of cosmic-ray data, followed by the fast and complete recovery of the initial gas properties using a lightweight gas circulation and purification system.

Dysregulation of lysosomal morphology by pathogenic LRRK2 is corrected by TPC2 inhibition

PubMed Central

Hockey, Leanne N.; Kilpatrick, Bethan S.; Eden, Emily R.; Lin-Moshier, Yaping; Brailoiu, G. Cristina; Brailoiu, Eugen; Futter, Clare E.; Schapira, Anthony H.; Marchant, Jonathan S.; Patel, Sandip

2015-01-01

ABSTRACT Two-pore channels (TPCs) are endolysosomal ion channels implicated in Ca2+ signalling from acidic organelles. The relevance of these ubiquitous proteins for human disease, however, is unclear. Here, we report that lysosomes are enlarged and aggregated in fibroblasts from Parkinson disease patients with the common G2019S mutation in LRRK2. Defects were corrected by molecular silencing of TPC2, pharmacological inhibition of TPC regulators [Rab7, NAADP and PtdIns(3,5)P2] and buffering local Ca2+ increases. NAADP-evoked Ca2+ signals were exaggerated in diseased cells. TPC2 is thus a potential drug target within a pathogenic LRRK2 cascade that disrupts Ca2+-dependent trafficking in Parkinson disease. PMID:25416817

Fibronectins containing extradomain A or B enhance osteoblast differentiation via distinct integrins

PubMed Central

Sens, Carla; Huck, Katrin; Pettera, Stefan; Uebel, Stephan; Wabnitz, Guido; Moser, Markus; Nakchbandi, Inaam A.

2017-01-01

Fibronectin is a multidomain protein secreted by various cell types. It forms a network of fibers within the extracellular matrix and impacts intracellular processes by binding to various molecules, primarily integrin receptors on the cells. Both the presence of several isoforms and the ability of the various domains and isoforms to bind to a variety of integrins result in a wide range of effects. In vivo findings suggest that fibronectin isoforms produced by the osteoblasts enhance their differentiation. Here we report that the isoform characterized by the presence of extradomain A activates α4β1 integrin and augments osteoblast differentiation. In addition, the isoform containing extradomain B enhances the binding of fibronectin through the RGD sequence to β3-containing integrin, resulting in increased mineralization by and differentiation of osteoblasts. Our study thus reveals novel functions for two fibronectin isoforms and the mediating receptors in osteoblast differentiation. PMID:28325836

Channel formation by serum amyloid A: a potential mechanism for amyloid pathogenesis and host defense.

PubMed

Hirakura, Yutaka; Carreras, Isabel; Sipe, Jean D; Kagan, Bruce L

2002-03-01

Serum amyloid A (SAA) is a family of closely related apolipoproteins associated with high density lipoprotein (HDL). Subclasses of SAA isoforms are differentially expressed constitutively and during inflammation. During states of infection or inflammation, levels of HDL bound, acute phase isoforms of SAA rise as much as 1000-fold in the serum, suggesting that it might play a role in host defense. Following recurrent or chronic inflammation, an N-terminal peptide fragment of SAA known as amyloid A (AA) assembles into fibrils causing extensive damage to spleen, liver, and kidney, and rapidly progressing to death. In the present paper, we report the novel finding that a recombinant acute phase isoform variant of human SAA 1.1 (SAAp) readily forms ion-channels in planar lipid bilayer membranes at physiologic concentrations. These channels are voltage-independent, poorly selective, and are relatively long-lived This type of channel would place a severe metabolic strain on various kinds of cells. Expression of human SAA 1.1 in bacteria induces lysis of bacterial cells, while expression of the constitutive isoform (human SAA4) does not. Secondary structural analysis of the SAA isoforms in dicates a strong hydrophobicity of the N-terminal of the acute phase isoform relative to the constitutive SAA4 isoform, which may be responsible for the bactericidal activity of the former, in keeping with the notion that SAA 1 targets cell membranes and forms channels in them. Channel formation may thus be related to a host defense role of acute phase SAA isoforms and may also be the mechanism of end organ damage in AA and other amyloidoses.

Comparison of active cooling devices to passive cooling for rehabilitation of firefighters performing exercise in thermal protective clothing: A report from the Fireground Rehab Evaluation (FIRE) trial

PubMed Central

Hostler, David; Reis, Steven E; Bednez, James C; Kerin, Sarah; Suyama, Joe

2010-01-01

Background Thermal protective clothing (TPC) worn by firefighters provides considerable protection from the external environment during structural fire suppression. However, TPC is associated with physiological derangements that may have adverse cardiovascular consequences. These derangements should be treated during on-scene rehabilitation periods. Objective The present study examined heart rate and core temperature responses during the application of four active cooling devices, currently being marketed to the fire service for on-scene rehab, and compared them to passive cooling in a moderate temperature (approximately 24°C) and to an infusion of cold (4°C) saline. Methods Subjects exercised in TPC in a heated room. Following an initial exercise period (BOUT 1) the subjects exited the room, removed TPC, and for 20 minutes cooled passively at room temperature, received an infusion of cold normal saline, or were cooled by one of four devices (fan, forearm immersion in water, hand cooling, water perfused cooling vest). After cooling, subjects donned TPC and entered the heated room for another 50-minute exercise period (BOUT 2). Results Subjects were not able to fully recover core temperature during a 20-minute rehab period when provided rehydration and the opportunity to completely remove TPC. Exercise duration was shorter during BOUT 2 when compared to BOUT 1 but did not differ by cooling intervention. The overall magnitude and rate of cooling and heart rate recovery did not differ by intervention. Conclusions No clear advantage was identified when active cooling devices and cold intravenous saline were compared to passive cooling in a moderate temperature after treadmill exercise in TPC. PMID:20397868

Action of aluminum, novel TPC1-type channel inhibitor, against salicylate-induced and cold-shock-induced calcium influx in tobacco BY-2 cells.

PubMed

Lin, Cun; Yu, Yawei; Kadono, Takashi; Iwata, Michiaki; Umemura, Kenji; Furuichi, Takuya; Kuse, Masaki; Isobe, Minoru; Yamamoto, Yoko; Matsumoto, Hideaki; Yoshizuka, Kazuharu; Kawano, Tomonori

2005-07-08

Previously, effect of Al ions on calcium signaling was assessed in tobacco cells expressing a Ca2+-monitoring luminescent protein, aequorin and a newly isolated putative plant Ca2+ channel protein from Arabidopsis thaliana, AtTPC1 (two-pore channel 1). TPC1 channels were shown to be the only channel known to be sensitive to Al and they are responsive to reactive oxygen species and cryptogein, a fungal elicitor protein. Thus, involvement of TPC1 channels in calcium signaling leading to development of plant defense mechanism has been suggested. Then, the use of Al as a specific inhibitor of TPC1-type plant calcium channels has been proposed. Here, using transgenic tobacco BY-2 cells expressing aequorin, we report on the evidence in support of the involvement of Al-sensitive signaling pathway requiring TPC1-type channel-dependent Ca2+ influx in response to salicylic acid, a key plant defense-inducing agent, but not to an elicitor prepared from the cell wall of rice blast disease fungus Magnaporthe grisea. In addition, involvement of Al-sensitive Ca2+ channels in response to cold shock was also tested. The data suggested that the elicitor used here induces the Ca2+ influx via Al-insensitive path, while salicylic acid and cold-shock-stimulate the influx of Ca2+ via Al-sensitive mechanism.

Convergent regulation of the lysosomal two-pore channel-2 by Mg2+, NAADP, PI(3,5)P2 and multiple protein kinases

PubMed Central

Jha, Archana; Ahuja, Malini; Patel, Sandip; Brailoiu, Eugen; Muallem, Shmuel

2014-01-01

Lysosomal Ca2+ homeostasis is implicated in disease and controls many lysosomal functions. A key in understanding lysosomal Ca2+ signaling was the discovery of the two-pore channels (TPCs) and their potential activation by NAADP. Recent work concluded that the TPCs function as a PI(3,5)P2 activated channels regulated by mTORC1, but not by NAADP. Here, we identified Mg2+ and the MAPKs, JNK and P38 as novel regulators of TPC2. Cytoplasmic Mg2+ specifically inhibited TPC2 outward current, whereas lysosomal Mg2+ partially inhibited both outward and inward currents in a lysosomal lumen pH-dependent manner. Under controlled Mg2+, TPC2 is readily activated by NAADP with channel properties identical to those in response to PI(3,5)P2. Moreover, TPC2 is robustly regulated by P38 and JNK. Notably, NAADP-mediated Ca2+ release in intact cells is regulated by Mg2+, PI(3,5)P2, and P38/JNK kinases, thus paralleling regulation of TPC2 currents. Our data affirm a key role for TPC2 in NAADP-mediated Ca2+ signaling and link this pathway to Mg2+ homeostasis and MAP kinases, pointing to roles for lysosomal Ca2+ in cell growth, inflammation and cancer. PMID:24502975

Convergent regulation of the lysosomal two-pore channel-2 by Mg²⁺, NAADP, PI(3,5)P₂ and multiple protein kinases.

PubMed

Jha, Archana; Ahuja, Malini; Patel, Sandip; Brailoiu, Eugen; Muallem, Shmuel

2014-03-03

Lysosomal Ca(2+) homeostasis is implicated in disease and controls many lysosomal functions. A key in understanding lysosomal Ca(2+) signaling was the discovery of the two-pore channels (TPCs) and their potential activation by NAADP. Recent work concluded that the TPCs function as a PI(3,5)P2 activated channels regulated by mTORC1, but not by NAADP. Here, we identified Mg(2+) and the MAPKs, JNK and P38 as novel regulators of TPC2. Cytoplasmic Mg(2+) specifically inhibited TPC2 outward current, whereas lysosomal Mg(2+) partially inhibited both outward and inward currents in a lysosomal lumen pH-dependent manner. Under controlled Mg(2+), TPC2 is readily activated by NAADP with channel properties identical to those in response to PI(3,5)P2. Moreover, TPC2 is robustly regulated by P38 and JNK. Notably, NAADP-mediated Ca(2+) release in intact cells is regulated by Mg(2+), PI(3,5)P2, and P38/JNK kinases, thus paralleling regulation of TPC2 currents. Our data affirm a key role for TPC2 in NAADP-mediated Ca(2+) signaling and link this pathway to Mg(2+) homeostasis and MAP kinases, pointing to roles for lysosomal Ca(2+) in cell growth, inflammation and cancer.

Absence of Intracellular Ion Channels TPC1 and TPC2 Leads to Mature-Onset Obesity in Male Mice, Due to Impaired Lipid Availability for Thermogenesis in Brown Adipose Tissue

PubMed Central

Lear, Pamela V.; González-Touceda, David; Porteiro Couto, Begoña; Viaño, Patricia; Guymer, Vanessa; Remzova, Elena; Tunn, Ruth; Chalasani, Annapurna; García-Caballero, Tomás; Hargreaves, Iain P.; Tynan, Patricia W.; Christian, Helen C.; Nogueiras, Rubén

2015-01-01

Intracellular calcium-permeable channels have been implicated in thermogenic function of murine brown and brite/beige adipocytes, respectively transient receptor potential melastin-8 and transient receptor potential vanilloid-4. Because the endo-lysosomal two-pore channels (TPCs) have also been ascribed with metabolic functionality, we studied the effect of simultaneously knocking out TPC1 and TPC2 on body composition and energy balance in male mice fed a chow diet. Compared with wild-type mice, TPC1 and TPC2 double knockout (Tpcn1/2−/−) animals had a higher respiratory quotient and became obese between 6 and 9 months of age. Although food intake was unaltered, interscapular brown adipose tissue (BAT) maximal temperature and lean-mass adjusted oxygen consumption were lower in Tpcn1/2−/− than in wild type mice. Phosphorylated hormone-sensitive lipase expression, lipid density and expression of β-adrenergic receptors were also lower in Tpcn1/2−/− BAT, whereas mitochondrial respiratory chain function and uncoupling protein-1 expression remained intact. We conclude that Tpcn1/2−/− mice show mature-onset obesity due to reduced lipid availability and use, and a defect in β-adrenergic receptor signaling, leading to impaired thermogenic activity, in BAT. PMID:25545384

Treatment of table olive processing wastewaters using novel photomodified ultrafiltration membranes as first step for recovering phenolic compounds.

PubMed

Garcia-Ivars, Jorge; Iborra-Clar, Maria-Isabel; Alcaina-Miranda, Maria-Isabel; Mendoza-Roca, José-Antonio; Pastor-Alcañiz, Laura

2015-06-15

Table olive processing wastewaters (TOPW) have high salt concentration and total phenolic content (TPC) causing many environmental problems. To reduce them, ultrafiltration (UF) was applied for treating TOPW. However, NaCl, which is the main responsible of salinity in TOPW, and phenols are small molecules that cannot be separated by conventional UF membranes. They have serious problems caused by fouling, which can be overcome using membrane modification techniques. For these reasons, photomodification may be an effective technique to obtain a stream rich in TPC due to the changes in membrane surface properties. UV-modification in the presence of two hydrophilic compounds (polyethylene glycol and aluminium oxide) was performed to achieve membranes with high reductions of organic matter and to keep the TPC as high as possible. Commercial polyethersulfone (PES) membranes of 30 kDa were used. Surface modification was evaluated using FTIR-ATR spectroscopy and membrane performance was studied by calculating the rejection ratios of colour, chemical oxygen demand (COD) and TPC. Results demonstrated that UF is a useful pre-treatment to reduce organic matter from TOPW, obtaining a permeate rich in TPC. PES/Al2O3 membranes displayed superior antifouling properties and rejection values, keeping high the TPC (>95%). Therefore, UF using modified membranes is an appropriate and sustainable technique for treating TOPW. Copyright © 2015 Elsevier B.V. All rights reserved.

Antioxidant capacity and phenolics content of apricot (Prunus armeniaca L.) kernel as a function of genotype.

PubMed

Korekar, Girish; Stobdan, Tsering; Arora, Richa; Yadav, Ashish; Singh, Shashi Bala

2011-11-01

Fourteen apricot genotypes grown under similar cultural practices in Trans-Himalayan Ladakh region were studied to find out the influence of genotype on antioxidant capacity and total phenolic content (TPC) of apricot kernel. The kernels were found to be rich in TPC ranging from 92.2 to 162.1 mg gallic acid equivalent/100 g. The free radical-scavenging activity in terms of inhibitory concentration (IC(50)) ranged from 43.8 to 123.4 mg/ml and ferric reducing antioxidant potential (FRAP) from 154.1 to 243.6 FeSO(4).7H(2)O μg/ml. A variation of 1-1.7 fold in total phenolic content, 1-2.8 fold in IC(50) by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and 1-1.6 fold in ferric reducing antioxidant potential among the examined kernels underlines the important role played by genetic background for determining the phenolic content and antioxidant potential of apricot kernel. A positive significant correlation between TPC and FRAP (r=0.671) was found. No significant correlation was found between TPC and IC(50); FRAP and IC(50); TPC and physical properties of kernel. Principal component analysis demonstrated that genotypic effect is more pronounced towards TPC and total antioxidant capacity (TAC) content in apricot kernel while the contribution of seed and kernel physical properties are not highly significant.

Thick Filament Length and Isoform Composition Determine Self-Organized Contractile Units in Actomyosin Bundles

PubMed Central

Thoresen, Todd; Lenz, Martin; Gardel, Margaret L.

2013-01-01

Diverse myosin II isoforms regulate contractility of actomyosin bundles in disparate physiological processes by variations in both motor mechanochemistry and the extent to which motors are clustered into thick filaments. Although the role of mechanochemistry is well appreciated, the extent to which thick filament length regulates actomyosin contractility is unknown. Here, we study the contractility of minimal actomyosin bundles formed in vitro by mixtures of F-actin and thick filaments of nonmuscle, smooth, and skeletal muscle myosin isoforms with varied length. Diverse myosin II isoforms guide the self-organization of distinct contractile units within in vitro bundles with shortening rates similar to those of in vivo myofibrils and stress fibers. The tendency to form contractile units increases with the thick filament length, resulting in a bundle shortening rate proportional to the length of constituent myosin thick filament. We develop a model that describes our data, providing a framework in which to understand how diverse myosin II isoforms regulate the contractile behaviors of disordered actomyosin bundles found in muscle and nonmuscle cells. These experiments provide insight into physiological processes that use dynamic regulation of thick filament length, such as smooth muscle contraction. PMID:23442916

Comparative study of the effects of solid-state fermentation with three filamentous fungi on the total phenolics content (TPC), flavonoids, and antioxidant activities of subfractions from oats (Avena sativa L.).

PubMed

Cai, Shengbao; Wang, Ou; Wu, Wei; Zhu, Songjie; Zhou, Feng; Ji, Baoping; Gao, Fengyi; Zhang, Di; Liu, Jia; Cheng, Qian

2012-01-11

The aim of present work was to investigate the effect of solid-state fermentation with filamentous fungi (Aspergillus oryzae var. effuses, Aspergillus oryzae, and Aspergillus niger) on total phenolics content (TPC), flavonoids, and antioxidant activities of four subfractions of oat, namely, n-hexane, ethyl acetate (EA), n-butanol, and water, and compare them to their corresponding subfractions of unfermented oat. The TPC and total flavonoids increased dramatically, especially in EA subfractions (p < 0.05). The levels of antioxidant activity of subfractions were also significantly enhanced (p < 0.05). The highest antioxidant activities were also found in the EA subfractions. The polyphenols in EA were analyzed by high-performance liquid chromatography at 280 nm. Most polyphenols were increased remarkably, especially ferulic and caffeic acids. There was a clear correlation between the TPC and antioxidant activity. In conclusion, fungi fermentation is a potential bioprocess for increasing the TPC, flavonoids, and antioxidant activities of oat-based food.

Precise Nuclear Data Measurements Possible with the NIFFTE fissionTPC for Advanced Reactor Designs

NASA Astrophysics Data System (ADS)

Towell, Rusty; Niffte Collaboration

2015-10-01

The Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) Collaboration has applied the proven technology of Time Projection Chambers (TPC) to the task of precisely measuring fission cross sections. With the NIFFTE fission TPC, precise measurements have been made during the last year at the Los Alamos Neutron Science Center from both U-235 and Pu-239 targets. The exquisite tracking capabilities of this device allow the full reconstruction of charged particles produced by neutron beam induced fissions from a thin central target. The wealth of information gained from this approach will allow systematics to be controlled at the level of 1%. The fissionTPC performance will be presented. These results are critical to the development of advanced uranium-fueled reactors. However, there are clear advantages to developing thorium-fueled reactors such as Liquid Fluoride Thorium Reactors over uranium-fueled reactors. These advantages include improved reactor safety, minimizing radioactive waste, improved reactor efficiency, and enhanced proliferation resistance. The potential for using the fissionTPC to measure needed cross sections important to the development of thorium-fueled reactors will also be discussed.

Dysregulation of lysosomal morphology by pathogenic LRRK2 is corrected by TPC2 inhibition.

PubMed

Hockey, Leanne N; Kilpatrick, Bethan S; Eden, Emily R; Lin-Moshier, Yaping; Brailoiu, G Cristina; Brailoiu, Eugen; Futter, Clare E; Schapira, Anthony H; Marchant, Jonathan S; Patel, Sandip

2015-01-15

Two-pore channels (TPCs) are endolysosomal ion channels implicated in Ca(2+) signalling from acidic organelles. The relevance of these ubiquitous proteins for human disease, however, is unclear. Here, we report that lysosomes are enlarged and aggregated in fibroblasts from Parkinson disease patients with the common G2019S mutation in LRRK2. Defects were corrected by molecular silencing of TPC2, pharmacological inhibition of TPC regulators [Rab7, NAADP and PtdIns(3,5)P2] and buffering local Ca(2+) increases. NAADP-evoked Ca(2+) signals were exaggerated in diseased cells. TPC2 is thus a potential drug target within a pathogenic LRRK2 cascade that disrupts Ca(2+)-dependent trafficking in Parkinson disease. © 2015. Published by The Company of Biologists Ltd.

Third Party Collection Program; Case Study of Naval Hospital Oakland and Community Hospital of Monterey Peninsula

DTIC Science & Technology

1992-12-01

the reader to the hospital TPC program, the concept , the La:., and tbe progran implementation responsibilities. it qives a brief explanation of the DoD...Community Hospital of Monterey Peninsula (CHOMP). This thesis briefly introduces the reader to the hospital TPC program, the concept , the Law, and the...current program. E. THESIS CHAPTER SUMMARY The first chapter briefly introduces the reader to the hospital TPC program, the concept , the law, and the

Usefulness of tenderness to characterise fibromyalgia severity in women.

PubMed

Aparicio, V A; Carbonell-Baeza, A; Ortega, F B; Estevez, F; Ruiz, J R; Delgado-Fernández, M

2011-01-01

To investigate the usefulness of tenderness (tender points count (TPC) and algometer score) to characterise fibromyalgia (FM) severity and symptomatology in women. The study sample comprised 174 women aged 51±7 years. We ossesse tenderness using pressure algometry; quality of life by means of the Short-Form 36 Health Survey (SF-36) and the Hospital Anxiety and Depression Scale (HADS). We used the FM impact questionnaire (FIQ) to assess FM severity and symptomatology. Patients were categorised according to three FIQ-derived categories: FIQ<70 vs. ≥70; FIQ<59 vs. ≥59; and FM-type I and II. TPC was significantly higher in the group of patients with FIQ≥59 (16.9±2 vs. 15.6±4, p=0.02), whereas no differences between groups were observed according to FIQ≥70 (17.0±2 vs. 16.2±3, p=0.12) or FM type (16.8±3 for type II vs. 15.9±4 for type I, p=0.13). We observed a significant association between TPC and FIQ-job difficulty, pain, morning tiredness and stiffness dimensions (all p<0.05), yet it was not correlated with total score of FIQ, FIQ-anxiety, fatigue and depression dimensions (all p>0.05). Algometer score was lower in the FIQ≥70 (45.7±12 vs. 51.1±14, p=0.05) and FIQ≥59 (46.7±13 vs. 52.7±14, p=0.05) groups, and there were no difference between FM types (48.7±13 vs. 49.5±14 for type II and I respectively, p=0.81). Algometer score was not associated with total score of FIQ or FIQ dimensions (all p≥0.1). Widespread pain and pain hypersensitivity, as measured by TPC and algometer score, do not seem to be useful to characterise FM severity and symptomatology (measured by FIQ) in women.

IDENTIFICATION AND EXPRESSION ANALYSIS OF TWO 14-3-3 PROTEINS IN THE MOSQUITO Aedes aegypti, AN IMPORTANT ARBOVIRUSES VECTOR.

PubMed

Trujillo-Ocampo, Abel; Cázares-Raga, Febe Elena; Celestino-Montes, Antonio; Cortés-Martínez, Leticia; Rodríguez, Mario H; Hernández-Hernández, Fidel de la Cruz

2016-11-01

The 14-3-3 proteins are evolutionarily conserved acidic proteins that form a family with several isoforms in many cell types of plants and animals. In invertebrates, including dipteran and lepidopteran insects, only two isoforms have been reported. 14-3-3 proteins are scaffold molecules that form homo- or heterodimeric complexes, acting as molecular adaptors mediating phosphorylation-dependent interactions with signaling molecules involved in immunity, cell differentiation, cell cycle, proliferation, apoptosis, and cancer. Here, we describe the presence of two isoforms of 14-3-3 in the mosquito Aedes aegypti, the main vector of dengue, yellow fever, chikungunya, and zika viruses. Both isoforms have the conserved characteristics of the family: two protein signatures (PS1 and PS2), an annexin domain, three serine residues, targets for phosphorylation (positions 58, 184, and 233), necessary for their function, and nine alpha helix-forming segments. By sequence alignment and phylogenetic analysis, we found that the molecules correspond to Ɛ and ζ isoforms (Aeae14-3-3ε and Aeae14-3-3ζ). The messengers and protein products were present in all stages of the mosquito life cycle and all the tissues analyzed, with a small predominance of Aeae14-3-3ζ except in the midgut and ovaries of adult females. The 14-3-3 proteins in female midgut epithelial cells were located in the cytoplasm. Our results may provide insights to further investigate the functions of these proteins in mosquitoes. © 2016 Wiley Periodicals, Inc.

Theoretical and experimental studies of turbo product code with time diversity in free space optical communication.

PubMed

Han, Yaoqiang; Dang, Anhong; Ren, Yongxiong; Tang, Junxiong; Guo, Hong

2010-12-20

In free space optical communication (FSOC) systems, channel fading caused by atmospheric turbulence degrades the system performance seriously. However, channel coding combined with diversity techniques can be exploited to mitigate channel fading. In this paper, based on the experimental study of the channel fading effects, we propose to use turbo product code (TPC) as the channel coding scheme, which features good resistance to burst errors and no error floor. However, only channel coding cannot cope with burst errors caused by channel fading, interleaving is also used. We investigate the efficiency of interleaving for different interleaving depths, and then the optimum interleaving depth for TPC is also determined. Finally, an experimental study of TPC with interleaving is demonstrated, and we show that TPC with interleaving can significantly mitigate channel fading in FSOC systems.

Feasibility and Acceptability of Nurse-Led Primary Palliative Care for Older Adults with Chronic Conditions: A Pilot Study.

PubMed

Izumi, Shigeko Seiko; Basin, Basilia; Presley, Margo; McCalmont, Jean; Furuno, Jon P; Noble, Brie; Baggs, Judith G; Curtis, J Randall

2018-05-24

Many older adults live with serious illness for years before their death. Nurse-led primary palliative care could improve their quality of life and ability to stay in their community. To assess feasibility and acceptability of a nurse-led Transitional Palliative Care (TPC) program for older adults with serious illness. The study was a pilot trial of the TPC program in which registered nurses assisted patients with symptom management, communication with care providers, and advance care planning. Forty-one older adults with chronic conditions were enrolled in TPC or standard care groups. Feasibility was assessed through enrollment and attrition rates and degree of intervention execution. Acceptability was assessed through surveys and exit interviews with participants and intervention nurses. Enrollment rate for those approached was 68%, and completion rate for those enrolled was 71%. The TPC group found the intervention acceptable and helpful and was more satisfied with care received than the control group. However, one-third of participants perceived that TPC was more than they needed, despite the number of symptoms they experienced and the burdensomeness of their symptoms. More than half of the participants had little to no difficulty participating in daily activities. This study demonstrated that the nurse-led TPC program is feasible, acceptable, and perceived as helpful. However, further refinement is needed in selection criteria to identify the population who would most benefit from primary palliative care before future test of the efficacy of this intervention.

A total pleural covering of absorbable cellulose mesh prevents pneumothorax recurrence in patients with Birt-Hogg-Dubé syndrome.

PubMed

Mizobuchi, Teruaki; Kurihara, Masatoshi; Ebana, Hiroki; Yamanaka, Sumitaka; Kataoka, Hideyuki; Okamoto, Shouichi; Kobayashi, Etsuko; Kumasaka, Toshio; Seyama, Kuniaki

2018-05-15

Birt-Hogg-Dubé syndrome (BHDS) is a recently recognized inherited multiple cystic lung disease causing recurrent pneumothoraces. Similarly to the lesions in patients with lymphangioleiomyomatosis (LAM), the pulmonary cysts are innumerable and widely dispersed and cannot all be removed. We recently described a total pleural covering (TPC) that covers the entire visceral pleura with oxidized regenerated cellulose (ORC) mesh. TPC successfully prevented the recurrence of pneumothorax in LAM patients. The purpose of this study was to evaluate the effect of an ORC pleural covering on pneumothorax recurrence in BHDS patients. This retrospective study enrolled a total of 81 pneumothorax patients with the diagnosis of BHDS who underwent 90 covering surgeries from January 2010 to August 2017 at Tamagawa Hospital. During the first half of the study period, a lower pleural covering (LPC) which covered the affected area with ORC mesh was mainly used to treat 38 pneumothoraces. During the second half of the study period, TPC was primarily performed for 52 pneumothoraces. All the thoracoscopic surgeries were successfully performed without serious complications (≥ Clavien-Dindo grade III). The median follow-up periods after LPC/TPC were 66/34 months, respectively. Pneumothorax recurrence rates after LPC at 2.5/5/7.5 years postoperatively were 5.4/12/42%, respectively; none of the patients who had underwent TPC developed postoperative pneumothorax recurrence (P = 0.032). TPC might be an effective option for surgical treatment of intractable pneumothorax in patients with BHDS.

TPC2 polymorphisms associated with a hair pigmentation phenotype in humans result in gain of channel function by independent mechanisms.

PubMed

Chao, Yu-Kai; Schludi, Verena; Chen, Cheng-Chang; Butz, Elisabeth; Nguyen, O N Phuong; Müller, Martin; Krüger, Jens; Kammerbauer, Claudia; Ben-Johny, Manu; Vollmar, Angelika M; Berking, Carola; Biel, Martin; Wahl-Schott, Christian A; Grimm, Christian

2017-10-10

Two-pore channels (TPCs) are endolysosomal cation channels. Two members exist in humans, TPC1 and TPC2. Functional roles associated with the ubiquitously expressed TPCs include VEGF-induced neoangiogenesis, LDL-cholesterol trafficking and degradation, physical endurance under fasting conditions, autophagy regulation, the acrosome reaction in sperm, cancer cell migration, and intracellular trafficking of pathogens such as Ebola virus or bacterial toxins (e.g., cholera toxin). In a genome-wide association study for variants associated with human pigmentation characteristics two coding variants of TPC2, rs35264875 (encoding M484L) and rs3829241 (encoding G734E), have been found to be associated with a shift from brown to blond hair color. In two recent follow-up studies a role for TPC2 in pigmentation has been further confirmed. However, these human polymorphic variants have not been functionally characterized until now. The development of endolysosomal patch-clamp techniques has made it possible to investigate directly ion channel activities and characteristics in isolated endolysosomal organelles. We applied this technique here to scrutinize channel characteristics of the polymorphic TPC2 variants in direct comparison with WT. We found that both polymorphisms lead to a gain of channel function by independent mechanisms. We next conducted a clinical study with more than 100 blond- and brown/black-haired individuals. We performed a genotype/phenotype analysis and subsequently isolated fibroblasts from WT and polymorphic variant carriers for endolysosomal patch-clamp experimentation to confirm key in vitro findings.

WhichP450: a multi-class categorical model to predict the major metabolising CYP450 isoform for a compound

NASA Astrophysics Data System (ADS)

Hunt, Peter A.; Segall, Matthew D.; Tyzack, Jonathan D.

2018-02-01

In the development of novel pharmaceuticals, the knowledge of how many, and which, Cytochrome P450 isoforms are involved in the phase I metabolism of a compound is important. Potential problems can arise if a compound is metabolised predominantly by a single isoform in terms of drug-drug interactions or genetic polymorphisms that would lead to variations in exposure in the general population. Combined with models of regioselectivities of metabolism by each isoform, such a model would also aid in the prediction of the metabolites likely to be formed by P450-mediated metabolism. We describe the generation of a multi-class random forest model to predict which, out of a list of the seven leading Cytochrome P450 isoforms, would be the major metabolising isoforms for a novel compound. The model has a 76% success rate with a top-1 criterion and an 88% success rate for a top-2 criterion and shows significant enrichment over randomised models.

A human Polycomb isoform lacking the Pc box does not participate to PRC1 complexes but forms protein assemblies and represses transcription.

PubMed

Völkel, Pamela; Le Faou, Perrine; Vandamme, Julien; Pira, Dorcas; Angrand, Pierre-Olivier

2012-05-01

Polycomb repression controls the expression of hundreds of genes involved in development and is mediated by essentially two classes of chromatin-associated protein complexes. The Polycomb repressive complex 2 (PRC2) trimethylates histone H3 at lysine 27, an epigenetic mark that serves as a docking site for the PRC1 protein complex. Drosophila core PRC1 is composed of four subunits: Polycomb (Pc), Posterior sex combs (Psc), Polyhomeotic (Ph) and Sex combs extra (Sce). Each of these proteins has multiple orthologs in vertebrates, thus generating an enormous scope for potential combinatorial diversity. In particular, mammalian genomes encode five Pc family members: CBX2, CBX4, CBX6, CBX7 and CBX8. To complicate matters further, distinct isoforms might arise from single genes. Here, we address the functional role of the two human CBX2 isoforms. Owing to different polyadenylation sites and alternative splicing events, the human CBX2 locus produces two transcripts: a 5-exon transcript that encodes the 532-amino acid CBX2-1 isoform that contains the conserved chromodomain and Pc box and a 4-exon transcript encoding a shorter isoform, CBX2-2, lacking the Pc box but still possessing a chromodomain. Using biochemical approaches and a novel in vivo imaging assay, we show that the short CBX2-2 isoform lacking the Pc box, does not participate in PRC1 protein complexes, but self-associates in vivo and forms complexes of high molecular weight. Furthermore, the CBX2 short isoform is still able to repress transcription, suggesting that Polycomb repression might occur in the absence of PRC1 formation.

Thick filament length and isoform composition determine self-organized contractile units in actomyosin bundles.

PubMed

Thoresen, Todd; Lenz, Martin; Gardel, Margaret L

2013-02-05

Diverse myosin II isoforms regulate contractility of actomyosin bundles in disparate physiological processes by variations in both motor mechanochemistry and the extent to which motors are clustered into thick filaments. Although the role of mechanochemistry is well appreciated, the extent to which thick filament length regulates actomyosin contractility is unknown. Here, we study the contractility of minimal actomyosin bundles formed in vitro by mixtures of F-actin and thick filaments of nonmuscle, smooth, and skeletal muscle myosin isoforms with varied length. Diverse myosin II isoforms guide the self-organization of distinct contractile units within in vitro bundles with shortening rates similar to those of in vivo myofibrils and stress fibers. The tendency to form contractile units increases with the thick filament length, resulting in a bundle shortening rate proportional to the length of constituent myosin thick filament. We develop a model that describes our data, providing a framework in which to understand how diverse myosin II isoforms regulate the contractile behaviors of disordered actomyosin bundles found in muscle and nonmuscle cells. These experiments provide insight into physiological processes that use dynamic regulation of thick filament length, such as smooth muscle contraction. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Investigating Premature Ignition of Thruster Pressure Cartridges by Vibration-Induced Electrostatic Discharge

NASA Technical Reports Server (NTRS)

Woods, Stephen S.; Saulsberry, Regor

2010-01-01

Pyrotechnic thruster pressure cartridges (TPCs) are used for aeroshell separation on a new NASA crew launch vehicle. Nondestructive evaluation (NDE) during TPC acceptance testing indicated that internal assemblies moved during shock and vibration testing due to an internal bond anomaly. This caused concerns that the launch environment might produce the same movement and release propellant grains that might be prematurely ignited through impact or through electrostatic discharge (ESD) as grains vibrated against internal surfaces. Since a new lot could not be fabricated in time, a determination had to be made as to whether the lot was acceptable to fly. This paper discusses the ESD evaluation and a separate paper addresses the impact problem. A challenge to straight forward assessment existed due to the unavailability of triboelectric data characterizing the static charging characteristics of the propellants within the TPC. The approach examined the physical limitations for charge buildup within the TPC system geometry and evaluated it for discharge under simulated vibrations used to qualify components for launch. A facsimile TPC was fabricated using SS 301 for the case and surrogate worst case materials for the propellants based on triboelectric data. System discharge behavior was evaluated by applying high voltage to the point of discharge in air and by placing worst case charge accumulations within the facsimile TPC and forcing discharge. The facsimile TPC contained simulated propellant grains and lycopodium, a well characterized indicator for static discharge in dust explosions, and was subjected to accelerations equivalent to the maximum accelerations possible during launch. The magnitude of charge generated within the facsimile TPC system was demonstrated to lie in a range of 100 to 10,000 times smaller than the spark energies measured to ignite propellant grains in industry standard discharge tests. The test apparatus, methodology, and results are described in this paper.

The two pore channel TPC2 is dispensable in pancreatic β-cells for normal Ca2+ dynamics and insulin secretion

PubMed Central

Cane, Matthew C.; Parrington, John; Rorsman, Patrik; Galione, Antony; Rutter, Guy A.

2016-01-01

Ca2+ signals are central to the stimulation of insulin secretion from pancreatic β-cells by glucose and other agents, including glucagon-like peptide-1 (GLP-1). Whilst Ca2+ influx through voltage-gated Ca2+ channels on the plasma membrane is a key trigger for glucose-stimulated secretion, mobilisation of Ca2+ from acidic stores has been implicated in the control of more localised Ca2+ changes and membrane potential. Nicotinic acid adenine dinucleotide phosphate (NAADP), generated in β-cells in response to high glucose, is a potent mobiliser of these stores, and has been proposed to act through two pore channels (TPC1 and TPC2, murine gene names Tpcn1 and Tpcn2). Whilst the role of TPC1 in the control of Ca2+ mobilisation and insulin secretion was recently confirmed, conflicting data exist for TPC2. Here, we used the selective and efficient deleter strain, Ins1Cre to achieve β-cell selective deletion of the Tpcn2 gene in mice. βTpcn2 KO mice displayed normal intraperitoneal and oral glucose tolerance, and glucose-stimulated Ca2+ dynamics and insulin secretion from islets were similarly normal. GLP-1-induced Ca2+ increases involved an increase in oscillation frequency from 4.35 to 4.84 per minute (p = 0.04) at 8 mM glucose, and this increase was unaffected by the absence of Tpcn2. The current data thus indicate that TPC2 is not absolutely required for normal glucose- or incretin-stimulated insulin secretion from the β-cell. Our findings suggest that TPC1, whose expression tended to increase in Tpcn2 null islets, might be sufficient to support normal Ca2+ dynamics in response to stimulation by nutrients or incretins. PMID:26769314

High Voltage Tests in the LUX-ZEPLIN System Test

NASA Astrophysics Data System (ADS)

Whitis, Thomas; Lux-Zeplin Collaboration

2016-03-01

The LUX-ZEPLIN (LZ) project is a dark matter direct detection experiment using liquid xenon. The detector is a time projection chamber (TPC) requiring the establishment of a large electric field inside of the detector in order to drift ionization electrons. Historically, many xenon TPC designs have been unable to reach their design fields due to light production and breakdown. The LZ System Test is scaled so that with a cathode voltage of -50 kV, it will have the fields that will be seen in the LZ detector at -100 kV. It will use a fully instrumented but scaled-down version of the LZ TPC design with a vessel set and gas system designed for quick turnaround, allowing for iterative modifications to the TPC prototype and instrumentation. This talk will present results from the high voltage tests performed during the first runs of the LZ System Test.

A thiourea derivative as potential ionophore for copper sensing

NASA Astrophysics Data System (ADS)

Ying, Kook Shih; Heng, Lee Yook; Hassan, Nurul Izzaty; Hasbullah, Siti Aishah

2018-04-01

A new thiourea derivative, N1,N3-bis[[3,5-bis(trifluoromethyl)phenyl]carbamothioyl]isophthalamide (TPC), as a potential copper ionophore was investigated. TPC was immobilized via drop casting method into poly(n-butyl acrylate) pBA membrane and the sensor was characterized by potentiometric method. The sensor fabricated based on TPC showed a Nernstian response towards copper ion with the slope of 27.07±2.84 mV/decade in the range of 1.0×10-6 - 1.0-10-4 M and limit of detection of 6.24 × 10-7 M. In addition, based on the separate solution method (SSM), the logarithm selectivity coefficients were less than -3.00 for monovalent, divalent and trivalent cations that are present in the environmental water samples such as K+, Ca2+, Mg2+ and Fe3+. This confirmed that the sensor fabricated with TPC exhibited good sensitivity and selectivity towards copper ion.

mTor Regulates Lysosomal ATP-sensitive Two-Pore Na+ Channel to Adapt to Metabolic State

PubMed Central

Navarro, Betsy; Seo, Young-jun; Aranda, Kimberly; Shi, Lucy; Battaglia-Hsu, Shyuefang; Nissim, Itzhak; Clapham, David E.; Ren, Dejian

2014-01-01

SUMMARY Survival in the wild requires organismal adaptations to the availability of nutrients. Endosomes and lysosomes are key intracellular organelles that couple nutrition and metabolic status to cellular responses, but how they detect cytosolic ATP levels is not well understood. Here we identify an endolysosomal ATP-sensitive Na+ channel (lysoNaATP). The channel is a complex formed by Two-Pore Channels (TPC1 and TPC2), ion channels previously thought to be gated by nicotinic acid adenine dinucleotide phosphate (NAADP), and the mammalian target of rapamycin (mTOR). The channel complex detects nutrient status, becomes constitutively open upon nutrient removal and mTOR translocation off the lysosomal membrane, and controls the lysosome's membrane potential, pH stability, and the amino acid homeostasis. Mutant mice lacking lysoNaATP have much reduced exercise endurance after fasting. Thus, TPCs are a new ion channel family that couple the cell's metabolic state to endolysosomal function and are crucial for physical endurance during food restriction. PMID:23394946

Effect of incorporating finger millet in wheat flour on mixolab behavior, chapatti quality and starch digestibility.

PubMed

Sharma, Bharati; Gujral, Hardeep Singh; Solah, Vicky

2017-09-15

Wheat and finger millet flour (two cultivars) were blended in the ratio (3:1) to form a composite flour and its dough properties were studied on the mixolab. The chapatti making and digestibility behavior of the composite flour was also investigated. The wheat finger millet (WFM) flour blend displayed up to 30.7% higher total phenolic content (TPC), 38.2% higher total flavonoid content (TFC) and 75.4% higher antioxidant activity (AOA) than the wheat flour. Chapattis prepared from the composite blends exhibited lower retrogradation as evident by the mixolab retrogradation index, higher values of soluble starch and soluble amylose in stored chapatti. The slowly digestible starch (SDS) correlated positively (R=0.816, p<0.05) with TPC and water absorption correlated positively (R=0.995, p<0.05) with damage starch content. The chapattis made from the composite flour had higher SDS and resistant starch (RS) values demonstrating potential as a food with functional characteristics. Copyright © 2017 Elsevier Ltd. All rights reserved.

A truncated, activin-induced Smad3 isoform acts as a transcriptional repressor of FSHβ expression in mouse pituitary

PubMed Central

Kim, So-Youn; Zhu, Jie; Woodruff, Teresa K.

2011-01-01

The receptor-regulated protein Smad3 is key player in the signaling cascade stimulated by the binding of activin to its cell surface receptor. Upon phosphorylation, Smad3 forms a heterocomplex with Smad2 and Smad4, translocates to the nucleus and acts as a transcriptional co-activator. We have identified a unique isoform of Smad3 that is expressed in mature pituitary gonadotropes. 5' RACE revealed that this truncated Smad3 isoform is transcribed from an ATG site within exon 4 and consists of 7 exons encoding half of the linker region and the MH2 region. In pituitary cells, the truncated Smad3 isoform was phosphorylated upon activin treatment, in a manner that was temporally distinct from the phosphorylation of full-length Smad3. Activin-induced phosphorylation of Smad3 and the truncated Smad3 isoform was blocked by both follistatin and siRNA-mediated knockdown of Smad3. The truncated Smad3 isoform antagonized Smad3-mediated, activin-responsive promoter activity. We propose that the pituitary gonadotrope contains an ultra-short, activin-responsive feedback loop utilizing two different isoforms of Smad3, one which acts as an agonist (Smad3) and another that acts as an intracrine antagonist (truncated Smad3 isoform) to regulate FSHβ production. PMID:21664424

Pteridine concentrations differ between insectary-reared and field-collected Anopheles albimanus mosquitoes of the same physiological age.

PubMed

Penilla, R P; Rodríguez, M H; López, A D; Viader-Salvadó, J M; Sánchez, C N

2002-09-01

Biopterin, isoxanthopterin and 6-pterincarboxylic acid were identified in the head of the malaria vector mosquito Anopheles albimanus Weidemann (Diptera: Culicidae) by HPLC. Total pteridine concentrations (TPC) were estimated in heads, body parts (BP: abdomen, legs and wings) and whole bodies of insectary-reared and field-collected females, by spectrofluorometry, to investigate whether they could be used for age determination. Pteridine concentrations diminished with age in both mosquito groups. TPC correlated with chronological age in insectary-reared sugar-fed females (heads: r2 = 0.35, BP: r2 = 0.34, P < 0.001), but lower correlation occurred in blood-fed females (heads: r2 = 0.22, BP: r2 = 0.27). TPC differed among females of the same age fed with blood at different times (P < 0.05), indicating that bloodmeals modify the diminution rate of pteridines with age. Nevertheless, a polynomial significant correlation was documented for TPC and the number of ovipositions (heads: r2 = 0.24, BP: r2 = 0.27, whole body: r2 = 0.52, P < 0.001) in insectary-reared mosquitoes. This correlation was lower in field-collected mosquitoes (heads: r2 = 0.14, BP: r2 = 0.10, P < 0.05), which showed a remarkable pteridine increase in one-parous females. The correlation of TPC in whole body with physiological age was much less (r2 = 0.03). These observations indicate that TPC determination by spectrofluorometry is not a reliable method to estimate the age of An. albimanus females from the feral population.

Comparative biomechanical evaluation of a pin-sleeve transfixation system in cadaveric calf metacarpal bones.

PubMed

Brianza, Stefano; Vogel, Susan; Rothstock, Stephan; Thalhauser, Martin; Desrochers, Andrè; Boure, Ludovic

2013-01-01

To compare proximal fragment displacement and the peri-implant strain using a pin-sleeve cast (PSC) system and a transfixation pin cast (TPC) system on a cadaveric calf metacarpal bone fracture model. Experimental. Cadaveric calf metacarpal bones (n = 6 pairs). Paired samples were instrumented with either the TPC or the PSC systems. Strain gauges were applied proximal to the transfixation implants and the bones encased in cast material. The distal part of the construct was removed to mimic an unstable distal comminuted fracture. Constructs were fixed to the material testing machine and initially loaded in axial compression in their elastic range to determine construct stiffness. Constructs were loaded cyclically with a sinusoidal curve that increased until failure. Variables compared statistically between constructs were the initial construct stiffness and, at given load points, the mean metacarpal axial displacement in loading and unloading condition and mean axial strain. Initial construct mean ± SD axial stiffness was not significantly different between constructs (PSC: 689 ± 258; TPC: 879 ± 306 N/mm). There was no significant difference between either investigated displacements of metacarpal bones transfixed with PSC and those transfixed with TPC at all load points. The PSC constructs had a significant decrease in the recorded mean strain (502 ± 340 μstrain) compared to the TPC construct (1738 ± 2218 μstrain). The PSC significantly reduced peri-implant strain with comparable axial displacement to the TPC in cadaveric calf metacarpal bones. © Copyright 2012 by The American College of Veterinary Surgeons.

In-depth characterization of breast cancer tumor-promoting cell transcriptome by RNA sequencing and microarrays

PubMed Central

Soldà, Giulia; Merlino, Giuseppe; Fina, Emanuela; Brini, Elena; Moles, Anna; Cappelletti, Vera; Daidone, Maria Grazia

2016-01-01

Numerous studies have reported the existence of tumor-promoting cells (TPC) with self-renewal potential and a relevant role in drug resistance. However, pathways and modifications involved in the maintenance of such tumor subpopulations are still only partially understood. Sequencing-based approaches offer the opportunity for a detailed study of TPC including their transcriptome modulation. Using microarrays and RNA sequencing approaches, we compared the transcriptional profiles of parental MCF7 breast cancer cells with MCF7-derived TPC (i.e. MCFS). Data were explored using different bioinformatic approaches, and major findings were experimentally validated. The different analytical pipelines (Lifescope and Cufflinks based) yielded similar although not identical results. RNA sequencing data partially overlapped microarray results and displayed a higher dynamic range, although overall the two approaches concordantly predicted pathway modifications. Several biological functions were altered in TPC, ranging from production of inflammatory cytokines (i.e., IL-8 and MCP-1) to proliferation and response to steroid hormones. More than 300 non-coding RNAs were defined as differentially expressed, and 2,471 potential splicing events were identified. A consensus signature of genes up-regulated in TPC was derived and was found to be significantly associated with insensitivity to fulvestrant in a public breast cancer patient dataset. Overall, we obtained a detailed portrait of the transcriptome of a breast cancer TPC line, highlighted the role of non-coding RNAs and differential splicing, and identified a gene signature with a potential as a context-specific biomarker in patients receiving endocrine treatment. PMID:26556871

Developments toward Understanding and Improving the Low Energy Measurement Capabilities of a Fission Time Projection Chamber

NASA Astrophysics Data System (ADS)

Bundgaard, Jeremy J.

Nuclear physicists have been recently called upon for new, high precision fission measurements to improve existing fission models, ultimately enabling engineers to design next generation reactors as well as guarding the nation's stockpile. In response, a resurgence in fission research is aimed at developing detectors to design and build new experiments to meet these needs. The Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) collaboration has developed the fission Time Projection Chamber (fissionTPC) to measure neutron induced fission with unprecedented precision. The fissionTPC is annually deployed to the Los Alamos Neutron Science Center LANSCE where it operates with a neutron beam passing axially through the drift volume, irradiating heavy actinide targets to induce fission. The fissionTPC was developed at the Lawrence Livermore National Laboratory's (LLNL) TPC lab, where it is tested with spontaneous fission (SF) from radioactive sources, typically 252Cf and 244Cm, to characterize detector response, improve performance, and evolve the design. One of the experiments relevant for both nuclear energy and nonproliferation is to measure the neutron induced fission of 239Pu, which exhibits a high alpha activity, generating a large unwanted background for the fission measurements. The ratio of alpha to fission present in our neutron induced fission measurement of 239Pu is on the same order of magnitude as the 244Cm alpha/SF branching ratio. The high alpha rate required the TPC to be triggering on fission signals during beam time and we set out to build a trigger system, which, using 244Cm to produce a similar alpha to fission ratio as 239Pu in the neutron beam, we successfully demonstrated the viability of this approach. The trigger design has been evolved for use in NIFFTE's current measurements at LANSCE. In addition to several hardware and software contributions in the development and operation of the fissionTPC, a central purpose of this thesis was also to develop analyses to demonstrate the fissionTPC's performance abilities/limitations in measuring the alpha/SF branching ratio of 252Cf and 244Cm. Our method results in benchmarking the fissionTPC's ability to produce a competitive alpha/SF ratio for 252Cf with sub-percent precision.

Changes of the phenolic compounds and antioxidant activities in germinated adlay seeds.

PubMed

Xu, Lei; Wang, Pei; Ali, Barkat; Yang, Na; Chen, Yisheng; Wu, Fengfeng; Xu, Xueming

2017-09-01

Over the years, germinated adlay products have been used as both food source and folk medicine. This study investigated the changes of total phenolic content (TPC), total flavonoid content (TFC), antioxidant activities, and phenolic acid profiles of adlay seed during germination. Results revealed that phenolic compounds and antioxidant activities varied with the germination stages. Germination significantly increased the free form phenolic and flavonoid contents by 112.5% and 168.3%, respectively. However, both of the bound form phenolic and flavonoid contents significantly decreased after germination. Phenolic acid compositions were quantified via HPLC analysis, and the levels of vanillic, p-coumaric, caffeic, hydroxybenzoic and protocatechuic acids in the free phenolic extracts were found to be significantly increased. The improvement of the free and total phenolic and flavonoid contents by the germination process led to a significant enhancement of the antioxidant activities (evaluated by the ABTS, FRAP and ORAC assays). The TPC showed the highest correlation with ORAC values (r = 0.9979). Germinated adlay had higher free and total phenolic and flavonoid contents, and antioxidant activities than ungerminated adlay. This study indicates that germinated adlay could be a promising functional food, more suitable for human consumption. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Increasing the antioxidant activity, total phenolic and flavonoid contents by optimizing the germination conditions of amaranth seeds.

PubMed

Perales-Sánchez, Janitzio X K; Reyes-Moreno, Cuauhtémoc; Gómez-Favela, Mario A; Milán-Carrillo, Jorge; Cuevas-Rodríguez, Edith O; Valdez-Ortiz, Angel; Gutiérrez-Dorado, Roberto

2014-09-01

The aim of this study was to optimize the germination conditions of amaranth seeds that would maximize the antioxidant activity (AoxA), total phenolic (TPC), and flavonoid (TFC) contents. To optimize the germination bioprocess, response surface methodology was applied over three response variables (AoxA, TPC, TFC). A central composite rotable experimental design with two factors [germination temperature (GT), 20-45 ºC; germination time (Gt), 14-120 h] in five levels was used; 13 treatments were generated. The amaranth seeds were soaked in distilled water (25 °C/6 h) before germination. The sprouts from each treatment were dried (50 °C/8 h), cooled, and ground to obtain germinated amaranth flours (GAF). The best combination of germination bioprocess variables for producing optimized GAF with the highest AoxA [21.56 mmol trolox equivalent (TE)/100 g sample, dw], TPC [247.63 mg gallic acid equivalent (GAE)/100 g sample, dw], and TFC [81.39 mg catechin equivalent (CAE)/100 g sample, dw] was GT = 30 ºC/Gt = 78 h. The germination bioprocess increased AoxA, TPC, and TFC in 300-470, 829, and 213%, respectively. The germination is an effective strategy to increase the TPC and TFC of amaranth seeds for enhancing functionality with improved antioxidant activity.

Performance Evaluation of the COBRA GEM for the Application of the TPC

NASA Astrophysics Data System (ADS)

Terasaki, Kohei; Hamagaki, Hideki; Gunji, Taku; Yamaguchi, Yorito

2014-09-01

Suppression of the back-drifting ions from avalanche region to drift space (IBF: Ion Backflow) is the key for a Time Projection Chamber (TPC) since IBF easily distorts the drift field. To suppress IBF, Gating Grid system is widely used for the TPC but this limits the data taking rate. Gas Electron Multiplier (GEM) has advantages in the reduction of IBF and high rate capability. By adopting GEM, it is possible to run a TPC continuously under high rate and high multiplicity conditions. Motivated by the study of IBF reduction for RICH with Thick COBRA, which has been developed by F. A. Amero et al., we developed COBRA GEMs for the application of a TPC. With a stack configuration, IBF reaches about 0.1 ~ 0.5%, which is ×5--10 better IBF than the standard GEMs. However, the measured energy resolution with COBRA is 20% (σ) and this is much worse than the resolution with standard GEMs. Measurement of long-time stability of gain indicates that gain of COBRA varies significantly due to charging up effect. Simulation studies based on Garfield++ are performed for understanding quantitatively the reasons of worse energy resolution and instability of gain. In this presentation, we will report the simulation studies together with the measured performance of the COBRA GEM.

Hsp70 Isoforms Are Essential for the Formation of Kaposi’s Sarcoma-Associated Herpesvirus Replication and Transcription Compartments

PubMed Central

Baquero-Pérez, Belinda; Whitehouse, Adrian

2015-01-01

Kaposi’s sarcoma-associated herpesvirus (KSHV) is an oncogenic herpesvirus associated with various AIDS-related malignancies. Like other herpesviruses, multiple processes required for KSHV lytic replication, including viral transcription, viral DNA synthesis and capsid assembly occur in virus-induced intranuclear structures, termed replication and transcription compartments (RTCs). Here we utilised a novel methodology, combining subcellular fractionation and quantitative proteomics, to identify cellular proteins which are recruited to KSHV-induced RTCs and thus play a key role in KSHV lytic replication. We show that several isoforms of the HSP70 chaperone family, Hsc70 and iHsp70, are redistributed from the cytoplasm into the nucleus coinciding with the initial formation of KSHV-induced RTCs. We demonstrate that nuclear chaperone foci are dynamic, initially forming adjacent to newly formed KSHV RTCs, however during later time points the chaperones move within KSHV RTCs and completely co-localise with actively replicating viral DNA. The functional significance of Hsp70 isoforms recruitment into KSHV RTCs was also examined using the specific Hsp70 isoform small molecule inhibitor, VER-155008. Intriguingly, results highlight an essential role of Hsp70 isoforms in the KSHV replication cycle independent of protein stability and maturation. Notably, inhibition of Hsp70 isoforms precluded KSHV RTC formation and RNA polymerase II (RNAPII) relocalisation to the viral genome leading to the abolishment of global KSHV transcription and subsequent viral protein synthesis and DNA replication. These new findings have revealed novel mechanisms that regulate KSHV lytic replication and highlight the potential of HSP70 inhibitors as novel antiviral agents. PMID:26587836

Proteomic Analysis of Parkin Isoforms Expression in Different Rat Brain Areas.

PubMed

D'Amico, Agata Grazia; Maugeri, Grazia; Reitano, Rita; Cavallaro, Sebastiano; D'Agata, Velia

2016-10-01

PARK2 gene's mutations are related to the familial form of juvenile Parkinsonism, also known as the autosomic recessive juvenile Parkinsonism. This gene encodes for parkin, a 465-amino acid protein. To date, a large number of parkin isoforms, generated by an alternative splicing mechanism, have been described. Currently, Gene Bank lists 27 rat PARK2 transcripts, which matches to 20 exclusive parkin alternative splice variants. Despite the existence of these isoforms, most of the studies carried out so far, have been focused only on the originally cloned parkin. In this work we have analyzed the expression profile of parkin isoforms in some rat brain areas including prefrontal cortex, hippocampus, substantia nigra and cerebellum. To discriminate among these isoforms, we detected their localization through the use of two antibodies that are able to identify different domains of the parkin canonical sequence. Our analysis has revealed that at least fourteen parkin isoforms are expressed in rat brain with a various distribution in the regions analyzed. Our study might help to elucidate the pathophysiological role of these proteins in the central nervous system.

THE PRESENCE OF A B SUBUNIT INCREASES SENSITIVITY OF SODIUM CHANNEL NAV1.3, BUT NOT NAV1.2, TO TYPE II PYRETHROIDS.

EPA Science Inventory

Voltage-sensitive sodium channels (VSSCs) are a primary target of pyrethroid insecticides. VSSCs are comprised of a pore-forming ¿ and auxillary ß subunits, and multiple isoforms of both subunit types exist. The sensitivity of different isoform combinations to pyrethroids has not...

Identification of a novel splice variant isoform of TREM-1 in human neutrophil granules1

PubMed Central

Baruah, Sankar; Keck, Kathy; Vrenios, Michelle; Pope, Marshall; Pearl, Merideth; Doerschug, Kevin; Klesney-Tait, Julia

2015-01-01

Triggering receptor expressed on myeloid cells-1 (TREM-1) is critical for inflammatory signal amplification. Humans have two forms of TREM-1: a membrane receptor (mbTREM-1), associated with the adaptor DAP12, and a soluble receptor detected at times of infection. The membrane receptor isoform acts synergistically with the TLR pathway to promote cytokine secretion and neutrophil migration while the soluble receptor functions as a counter regulatory molecule. In multiple models of sepsis, exogenous administration of soluble forms of TREM-1 attenuates inflammation and markedly improves survival. Despite intense interest in soluble TREM-1 both as a clinical predictor of survival and as a therapeutic tool, the origin of native soluble TREM-1 remains controversial. Utilizing human neutrophils, we identified a 15 kDa TREM-1 isoform in primary (azurophilic) and secondary (specific) granules. Mass spectrometric analysis, ELISA, and immunoblot confirm that the 15 kD protein is a novel splice variant of TREM-1 (TREM-1sv). Neutrophil stimulation with P. aeruginosa, LPS, or PAM(3)Cys4 resulted in degranulation and release of TREM-1sv. The addition of exogenous TREM-1sv inhibited TREM-1 receptor mediated proinflammatory cytokine production. Thus these data reveal that TREM-1 isoforms simultaneously activate and inhibit inflammation via the canonical membrane TREM-1 molecule and this newly discovered granular isoform, TREM-1sv. PMID:26561551

A truncated, activin-induced Smad3 isoform acts as a transcriptional repressor of FSHβ expression in mouse pituitary.

PubMed

Kim, So-Youn; Zhu, Jie; Woodruff, Teresa K

2011-08-06

The receptor-regulated protein Smad3 is key player in the signaling cascade stimulated by the binding of activin to its cell surface receptor. Upon phosphorylation, Smad3 forms a heterocomplex with Smad2 and Smad4, translocates to the nucleus and acts as a transcriptional co-activator. We have identified a unique isoform of Smad3 that is expressed in mature pituitary gonadotropes. 5' RACE revealed that this truncated Smad3 isoform is transcribed from an ATG site within exon 4 and consists of 7 exons encoding half of the linker region and the MH2 region. In pituitary cells, the truncated Smad3 isoform was phosphorylated upon activin treatment, in a manner that was temporally distinct from the phosphorylation of full-length Smad3. Activin-induced phosphorylation of Smad3 and the truncated Smad3 isoform was blocked by both follistatin and siRNA-mediated knockdown of Smad3. The truncated Smad3 isoform antagonized Smad3-mediated, activin-responsive promoter activity. We propose that the pituitary gonadotrope contains an ultra-short, activin-responsive feedback loop utilizing two different isoforms of Smad3, one which acts as an agonist (Smad3) and another that acts as an intracrine antagonist (truncated Smad3 isoform) to regulate FSHβ production. Copyright © 2011 Elsevier Ltd. All rights reserved.

A comparison of basal and eye-flush tears for the analysis of cat tear proteins.

PubMed

Petznick, Andrea; Evans, Margaret D M; Madigan, Michele C; Markoulli, Maria; Garrett, Qian; Sweeney, Deborah F

2011-02-01

To identify a rapid and effective tear collection method providing sufficient tear volume and total protein content (TPC) for analysis of individual proteins in cats. Domestic adult short-haired cats (12-37 months; 2.7-6.6 kg) were used in the study. Basal tears without stimulation and eye-flush tears after instillation of saline (10 μl) were collected using microcapillary tubes from animal eyes either unwounded control or wounded with 9-mm central epithelial debridement giving four groups with n = 3. Tear comparisons were based on total time and rate for tear collection, TPC using micro bicinchoninic acid (BCA), tear immunoglobulin A (IgA), total matrix-metalloproteinase (MMP)-9 concentration using sandwich enzyme-linked immunosorbent assay (ELISA) and MMP-9 activity. Eye-flush tears were collected significantly faster than basal tears in wounded eyes with higher rates for tear collection in unwounded control and wounded eyes. TPC was significantly lower in eye-flush tears compared to basal tears. The relative proportion of tear IgA normalized to TPC (% IgA of TPC) was not significantly different between basal and eye-flush tears. In unwounded control eyes, MMP-9 was slightly higher in eye-flush than in basal tears; activity of MMP-9 in both tear types was similar. In wounded eyes, eye-flush tears showed highest MMP-9 levels and activity on Day 1, which subsequently decreased to Day 7. MMP-9 activity in basal tears from wounded eyes did not display changes in expression. Eye-flush tears can be collected rapidly providing sufficient tear volume and TPC. This study also indicates that eye-flush tears may be more suitable than basal tears for the analysis of MMPs following corneal wounding. © 2011 The Authors. Acta Ophthalmologica © 2011 Acta Ophthalmologica Scandinavica Foundation.

Web-based oil immersion whole slide imaging increases efficiency and clinical team satisfaction in hematopathology tumor board

PubMed Central

Chen, Zhongchuan Will; Kohan, Jessica; Perkins, Sherrie L.; Hussong, Jerry W.; Salama, Mohamed E.

2014-01-01

Background: Whole slide imaging (WSI) is widely used for education and research, but is increasingly being used to streamline clinical workflow. We present our experience with regard to satisfaction and time utilization using oil immersion WSI for presentation of blood/marrow aspirate smears, core biopsies, and tissue sections in hematology/oncology tumor board/treatment planning conferences (TPC). Methods: Lymph nodes and bone marrow core biopsies were scanned at ×20 magnification and blood/marrow smears at 83X under oil immersion and uploaded to an online library with areas of interest to be displayed annotated digitally via web browser. Pathologist time required to prepare slides for scanning was compared to that required to prepare for microscope projection (MP). Time required to present cases during TPC was also compared. A 10-point evaluation survey was used to assess clinician satisfaction with each presentation method. Results: There was no significant difference in hematopathologist preparation time between WSI and MP. However, presentation time was significantly less for WSI compared to MP as selection and annotation of slides was done prior to TPC with WSI, enabling more efficient use of TPC presentation time. Survey results showed a significant increase in satisfaction by clinical attendees with regard to image quality, efficiency of presentation of pertinent findings, aid in clinical decision-making, and overall satisfaction regarding pathology presentation. A majority of respondents also noted decreased motion sickness with WSI. Conclusions: Whole slide imaging, particularly with the ability to use oil scanning, provides higher quality images compared to MP and significantly increases clinician satisfaction. WSI streamlines preparation for TPC by permitting prior slide selection, resulting in greater efficiency during TPC presentation. PMID:25379347

Cloning of Novel Isoforms of the Human Gli2 Oncogene and Their Activities To Enhance Tax-Dependent Transcription of the Human T-Cell Leukemia Virus Type 1 Genome

PubMed Central

Tanimura, Akira; Dan, Shingo; Yoshida, Mitsuaki

1998-01-01

The expression of human T-cell leukemia virus type 1 (HTLV-1) is activated by interaction of a viral transactivator protein, Tax, and cellular transcription factor, CREB (cyclic AMP response element binding protein), which bind to a 21-bp enhancer in the long terminal repeats (LTR). THP (Tax-helping protein) was previously determined to enhance the transactivation by Tax protein. Here we report novel forms of the human homolog of a member of the Gli oncogene family, Gli2 (also termed Gli2/THP), an extended form of a zinc finger protein, THP, which was described previously. Four possible isoforms (hGli2 α, β, γ, and δ) are formed by combinations of two independent alternative splicings, and all the isoforms could bind to a DNA motif, TRE2S, in the LTR. The longer isoforms, α and β, were abundantly expressed in various cell lines including HTLV-1-infected T-cell lines. Fusion proteins of the hGli2 isoforms with the DNA-binding domain of Gal4 activated transcription when the reporter contained a Gal4-binding site and one copy of the 21-bp sequence, to which CREB binds. This activation was observed only in the presence of Tax. The 21-bp sequence in the reporter was also essential for the activation. These results suggest that simultaneous binding of hGli2 and CREB to the respective sites in the reporter seems to be critical for Tax protein to activate transcription. Consequently, it is probable that the LTR can be regulated by two independent signals through hGli2 and CREB, since the LTR contains the 21-bp and TRE2S sequences in the vicinity. PMID:9557682

Two-Pore Channels: Catalyzers of Endolysosomal Transport and Function

PubMed Central

Grimm, Christian; Chen, Cheng-Chang; Wahl-Schott, Christian; Biel, Martin

2017-01-01

Two-pore channels (TPCs) have recently emerged as a novel class of non-selective cation channels in the endolysosomal system. There are two members in the human genome, TPC1 and TPC2. Studies with TPC knockout and knockdown models have revealed that these channels participate in the regulation of multiple endolysosomal trafficking pathways which when dysregulated can lead to or influence the development of a range of different diseases such as lysosomal storage, metabolic, or infectious diseases. TPCs have been demonstrated to be activated by different endogenous stimuli, PI(3,5)P2 and NAADP, and ATP has been found to block TPC activation via mTOR. Loss of TPCs can lead to obesity and hypercholesterolemia, and to a slow-down of intracellular virus and bacterial toxin trafficking, it can affect VEGF-induced neoangiogenesis, autophagy, human hair pigmentation or the acrosome reaction in sperm. Moreover, physiological roles of TPCs in cardiac myocytes and pancreatic β cells have been postulated. PMID:28223936

Slow Control System for the NIFFTE Collaboration TPC

NASA Astrophysics Data System (ADS)

Ringle, Erik; Niffte Collaboration Collaboration

2011-10-01

As world energy concerns continue to dominate public policy in the 21st century, the need for cleaner and more efficient nuclear power is necessary. In order to effectively design and implement plans for generation IV nuclear reactors, more accurate fission cross-section measurements are necessary. The Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) collaboration, in an effort to meet this need, has constructed a Time Projection Chamber (TPC) which aims to reduce the uncertainty of the fission cross-section to less than 1%. Using the Maximum Integration Data Acquisition System (MIDAS) framework, slow control measurements are integrated into a single interface to facilitate off-site monitoring. The Hart Scientific 1560 Black Stack will be used with two 2564 Thermistor Scanner Modules to monitor internal temperature of the TPC. A Prologix GPIB to Ethernet controller will be used to interface the hardware with MIDAS. This presentation will detail the design and implementation of the slow control system for the TPC. This work was supported by the U.S. Department of Energy Division of Energy Research.

DarkSide-50: A WIMP Search with a Two-phase Argon TPC

NASA Astrophysics Data System (ADS)

Meyers, P. D.; Agnes, P.; Alton, D.; Arisaka, K.; Back, H. O.; Baldin, B.; Biery, K.; Bonfini, G.; Bossa, M.; Brigatti, A.; Brodsky, J.; Budano, F.; Cadonati, L.; Calaprice, F.; Canci, N.; Candela, A.; Cao, H.; Cariello, M.; Cavalcante, P.; Chavarria, A.; Chepurnov, A.; Cocco, A. G.; Crippa, L.; DAngelo, D.; D'Incecco, M.; Davini, S.; De Deo, M.; Derbin, A.; Di Eusanio, F.; Di Pietro, G.; Edkins, E.; Empl, A.; Fan, A.; Fiorillo, G.; Fomenko, K.; Forster, G.; Franco, D.; Gabriele, F.; Galbiati, C.; Goretti, A.; Grandi, L.; Gromov, M.; Guan, M.; Guardincerri, Y.; Hackett, B.; Herner, K.; Humble, P.; Hungerford, E. V.; Ianni, Al.; Ianni, An.; Joliet, C.; Keeter, K.; Kendziora, C.; Kidner, S.; Kobychev, V.; Koh, G.; Korablev, D.; Korga, G.; Kurlej, A.; Li, P.; Loer, B.; Lombardi, P.; Love, C.; Ludhova, L.; Luitz, S.; Ma, Y.; Machulin, I.; Mandarano, A.; Mari, S.; Maricic, J.; Martoff, C. J.; Meregaglia, A.; Meroni, E.; Meyers, P. D.; Milincic, R.; Montanari, D.; Montuschi, M.; Monzani, M. E.; Mosteiro, P.; Mount, B.; Muratova, V.; Musico, P.; Nelson, A.; Okounkova, M.; Orsini, M.; Ortica, F.; Pagani, L.; Pallavicini, M.; Pantic, E.; Papp, L.; Parmeggiano, S.; Parsells, R.; Pelczar, K.; Pelliccia, N.; Perasso, S.; Perfetto, F.; Pocar, A.; Pordes, S.; Qian, H.; Randle, K.; Ranucci, G.; Razeto, A.; Reinhold, B.; Romani, A.; Rossi, B.; Rossi, N.; Rountree, S. D.; Sablone, D.; Saggese, P.; Saldanha, R.; Sands, W.; Segreto, E.; Semenov, D.; Shields, E.; Skorokhvatov, M.; Smirnov, O.; Sotnikov, A.; Suvarov, Y.; Tartaglia, R.; Tatarowicz, J.; Testera, G.; Tonazzo, A.; Unzhakov, E.; Vogelaar, R. B.; Wada, M.; Wang, H.; Wang, Y.; Watson, A.; Westerdale, R.; Wojcik, M.; Wright, A.; Xu, J.; Yang, C.; Yoo, J.; Zavatarelli, S.; Zuzel, G.

DarkSide-50 is a two phase argon TPC for direct dark matter detection which is installed at the Gran Sasso underground laboratory, Italy. DarkSide-50 has a 50-kg active volume and will make use of underground argon low in 39Ar. The TPC is installed inside an active neutron veto made with boron-loaded high radiopurity liquid scintillator. The neutron veto is installed inside a 1000 m3 water Cherenkov muon veto. The DarkSide-50 TPC and cryostat are assembled in two radon-free clean rooms to reduce radioactive contaminants. The overall design aims for a background free exposure after selection cuts are applied. The expected sensitivity for WIMP-nucleon cross section is of the order of 10-45 cm2 for WIMP masses around 100 GeV/c2. The commissioning and performance of the detector are described. Details of the low-radioactivity underground argon and other unique features of the projects are reported.

Ultrasound assisted extraction of polyphenols and their distribution in whole mung bean, hull and cotyledon.

PubMed

Singh, Barinderjit; Singh, Narpinder; Thakur, Sheetal; Kaur, Amritpal

2017-03-01

In this study, extraction of polyphenols using different solvents (acetone, ethanol, methanol and water) with ultrasound and conventional method from whole mung bean (WMB), hull and cotyledon was conducted. Total phenolic content (TPC), total flavonoids content (TFC), total antioxidant activities (TAA), ferric reducing power (FRP) and DPPH radical scavenging activity were determined. Ultrasound treated extracts exhibited higher TPC, TFC, TAA, FRP and DPPH in different mung bean fractions than CSE. Among the solvents, acetone showed better TPC, TFC, TAA, FRP and DPPH. Hull had significantly higher TPC, TFC, TAA, FRP and DPPH than WMB and cotyledon. Sinapic acid (SA) was the major polyphenol in different fractions. Acetone extract of hull showed high polyphenol content. SA, ferulic acid, catechin, p-coumaric acid, resveratrol, quercetin and luteolin were the major contributors to antioxidant activity of acetone extract. Mung bean hull contained the maximum polyphenols and acetone was observed to be the best extraction medium for polyphenols in combination with ultrasound.

Departure gate of acidic Ca2+ confirmed

PubMed Central

Jentsch, Thomas J; Hoegg-Beiler, Maja B; Vogt, Janis

2015-01-01

More potent, but less known than IP3 that liberates Ca2+ from the ER, NAADP releases Ca2+ from acidic stores. The notion that TPC channels mediate this Ca2+ release was questioned recently by studies suggesting that TPCs are rather PI(3,5)P2-activated Na+ channels. Ruas et al (2015) now partially reconcile these views by showing that TPCs significantly conduct both cations and confirm their activation by both NAADP and PI(3,5)P2. They attribute the failure of others to observe TPC-dependent NAADP-induced Ca2+ release in vivo to inadequate mouse models that retain partial TPC function. PMID:26022292

Development of time projection chamber for precise neutron lifetime measurement using pulsed cold neutron beams

NASA Astrophysics Data System (ADS)

Arimoto, Y.; Higashi, N.; Igarashi, Y.; Iwashita, Y.; Ino, T.; Katayama, R.; Kitaguchi, M.; Kitahara, R.; Matsumura, H.; Mishima, K.; Nagakura, N.; Oide, H.; Otono, H.; Sakakibara, R.; Shima, T.; Shimizu, H. M.; Sugino, T.; Sumi, N.; Sumino, H.; Taketani, K.; Tanaka, G.; Tanaka, M.; Tauchi, K.; Toyoda, A.; Tomita, T.; Yamada, T.; Yamashita, S.; Yokoyama, H.; Yoshioka, T.

2015-11-01

A new time projection chamber (TPC) was developed for neutron lifetime measurement using a pulsed cold neutron spallation source at the Japan Proton Accelerator Research Complex (J-PARC). Managing considerable background events from natural sources and the beam radioactivity is a challenging aspect of this measurement. To overcome this problem, the developed TPC has unprecedented features such as the use of polyether-ether-ketone plates in the support structure and internal surfaces covered with 6Li-enriched tiles to absorb outlier neutrons. In this paper, the design and performance of the new TPC are reported in detail.

High-pressure xenon time projection Titanium chamber: a methodology for detecting background radiation in neutrinoless double-beta decay experiments

NASA Astrophysics Data System (ADS)

Bachri, A.; Elmhamdi, A.; Hawron, M.; Grant, P.; Zazoum, B.; Martin, C.

2017-10-01

The xenon time projection chamber (TPC) promises a novel detection method for neutrinoless double-beta decay (0ν β β ) experiments. The TPC is capable of discovering the rare 0ν β β ionization signal of a distinct topological signature, with a decay energy Qββ = 2.458 MeV . However, more frequent internal (within TPC) and external events are also capable of depositing energy in the range of the Qβ β -value inside the chamber, thus mimicking 0ν β β or interfering with its direct observation. In the following paper, we illustrate a methodology for background radiation evaluation, assuming a basic cylindrical design for a toy titanium TPC that is capable of containing 100 kg of xenon gas at 20 atm pressure; we estimate the background budget and analyze the most prominent problematic events via theoretical calculation. Gamma rays emitted from nuclei of 214Bi and 208Tl present in the outer-shell titanium housing of the TPC are an example of such events for which we calculate probabilities of occurrences. We also study the effect of alpha-neutron (α-n)-induced neutrons and calculate their rate. Alpha particles which are created by the decay of naturally occurring uranium and thorium present in most materials, can react with the nucleus of low Z elements, prompting the release of neutrons and leading to thermal neutron capture. Our calculations suggest that the typical polytetrafluoroethylene (PTFE) inner coating of the chamber would constitute the primary material for neutron production, specifically; we find that the fluorine component of Teflon is much more likely to undergo an (α-n) reaction. From known contamination, we calculate an alpha production rate to be 5.5 × 107 alpha/year for the highest-purity titanium vessel with a Teflon lining. Lastly, using measurements of neutron flux from alpha bombardment, we estimate the expected neutron flux from the materials of the proposed toy TPC and identify all gamma rays (prompt or delayed, of energies comparable to the Qβ β -value) originating from thermal neutron capture with all stable elemental isotopes present in the TPC. We show that to limit the most probable reactions to a rate of one event per year or less, the neutron flux would have to be reduced to (3-6) × 10-10 cm-2ṡs-1. The predictions of our crude theoretical calculation are in good agreement with full simulation of TPC radiation background by existing experimental collaboration using xenon for 0ν β β experiment.

Reactivity of thyroid papillary carcinoma cells to thyroid stimulating hormone-dominated endocrine therapy

PubMed Central

Ma, Yuqin; Zhang, Xia; Wang, Yutao

2017-01-01

This study investigated the effect of thyroid stimulating hormone (TSH) on the proliferation of papillary thyroid carcinoma (PTC) cells and the therapeutic effect of levothyroxine sodium (TH). PTC cells (TPC-1) were cultured using 0.1, 1.0 and 10 U/l TSH and 10−2, 10−4 and 10−6 mol/l TH. After the appropriate concentration was screened, TPC-1 cells were further divided into control group, TSH group, TH group and TSH+TH group. The cell proliferation was detected via methyl thiazolyl tetrazolium (MTT) method, TPC-1 cell cycle was detected via flow cytometer, and the mRNA and protein expression of cyclin D1 were detected via real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Compared with control group, TSH significantly promoted the proliferation of TPC-1 cells (P<0.05 or P<0.01), obviously promoted the transition of TPC-1 cells from G1 phase to S phase (P<0.01) and remarkably increased the mRNA and protein expression of cyclin D1 (P<0.01); but TH had a significant inhibitory effect on these results of TSH (P<0.05 or P<0.01). TSH can promote the proliferation of PTC cells, and the appropriate complement of TH can inhibit its proliferation. PMID:29250166

Efficient ECH-assisted plasma start-up using trapped particle configuration in the versatile experiment spherical torus

NASA Astrophysics Data System (ADS)

An, YoungHwa; Lee, Jeongwon; Jo, JongGab; Jung, Bong-Ki; Lee, HyunYeong; Chung, Kyoung-Jae; Na, Yong-Su; Hahm, T. S.; Hwang, Y. S.

2017-01-01

An efficient and robust ECH (electron cyclotron heating)-assisted plasma start-up scheme with a low loop voltage and low volt-second consumption utilizing the trapped particle configuration (TPC) has been developed in the versatile experiment spherical torus (VEST). The TPC is a mirror-like magnetic field configuration providing a vertical magnetic field in the same direction as the equilibrium field. It significantly enhances ECH pre-ionization with enhanced particle confinement due to its mirror effect, and intrinsically provides an equilibrium field with a stable decay index enabling prompt plasma current initiation. Consequently, the formation of TPC before the onset of the loop voltage allows the plasma to start up with a lower loop voltage and lower volt-second consumption as well as a wider operation range in terms of ECH pre-ionization power and H2 filling pressure. The TPC can improve the widely-used field null configuration significantly for more efficient start-up when ECH pre-ionization is used. This can then be utilized in superconducting tokamaks requiring a low loop voltage start-up, such as ITER, or in spherical tori with limited volt-seconds. The TPC can be particularly useful in superconducting tokamaks with a limited current slew-rate of superconducting PF coils, as it can save volt-second consumption before plasma current initiation by providing prompt initiation with an intrinsic stable equilibrium field.

The Na, K-ATPase β-Subunit Isoforms Expression in Glioblastoma Multiforme: Moonlighting Roles

PubMed Central

Rotoli, Deborah; Cejas, Mariana-Mayela; Maeso, María-del-Carmen; Pérez-Rodríguez, Natalia-Dolores; Morales, Manuel; Ávila, Julio

2017-01-01

Glioblastoma multiforme (GBM) is the most common form of malignant glioma. Recent studies point out that gliomas exploit ion channels and transporters, including Na, K-ATPase, to sustain their singular growth and invasion as they invade the brain parenchyma. Moreover, the different isoforms of the β-subunit of Na, K-ATPase have been implicated in regulating cellular dynamics, particularly during cancer progression. The aim of this study was to determine the Na, K-ATPase β subunit isoform subcellular expression patterns in all cell types responsible for microenvironment heterogeneity of GBM using immunohistochemical analysis. All three isoforms, β1, β2/AMOG (Adhesion Molecule On Glia) and β3, were found to be expressed in GBM samples. Generally, β1 isoform was not expressed by astrocytes, in both primary and secondary GBM, although other cell types (endothelial cells, pericytes, telocytes, macrophages) did express this isoform. β2/AMOG and β3 positive expression was observed in the cytoplasm, membrane and nuclear envelope of astrocytes and GFAP (Glial Fibrillary Acidic Protein) negative cells. Interestingly, differences in isoforms expression have been observed between primary and secondary GBM: in secondary GBM, β2 isoform expression in astrocytes was lower than that observed in primary GBM, while the expression of the β3 subunit was more intense. These changes in β subunit isoforms expression in GBM could be related to a different ionic handling, to a different relationship between astrocyte and neuron (β2/AMOG) and to changes in the moonlighting roles of Na, K-ATPase β subunits as adaptor proteins and transcription factors. PMID:29117147

Tocotrienols suppress proinflammatory markers and cyclooxygenase-2 expression in RAW264.7 macrophages.

PubMed

Yam, Mun-Li; Abdul Hafid, Sitti Rahma; Cheng, Hwee-Ming; Nesaretnam, Kalanithi

2009-09-01

Tocotrienols are powerful chain breaking antioxidant. Moreover, they are now known to exhibit various non-antioxidant properties such as anti-cancer, neuroprotective and hypocholesterolemic functions. This study was undertaken to investigate the anti-inflammatory effects of tocotrienol-rich fraction (TRF) and individual tocotrienol isoforms namely delta-, gamma-, and alpha-tocotrienol on lipopolysaccharide-stimulated RAW264.7 macrophages. The widely studied vitamin E form, alpha-tocopherol, was used as comparison. Stimulation of RAW264.7 with lipopolysaccharide induced the release of various inflammatory markers. 10 mcirog/ml of TRF and all tocotrienol isoforms significantly inhibited the production of interleukin-6 and nitric oxide. However, only alpha-tocotrienol demonstrated a significant effect in lowering tumor necrosis factor-alpha production. Besides, TRF and all tocotrienol isoforms except gamma-tocotrienol reduced prostaglandin E(2) release. It was accompanied by the down-regulation of cyclooxygenase-2 gene expression by all vitamin E forms except alpha-tocopherol. Collectively, the data suggested that tocotrienols are better anti-inflammatory agents than alpha-tocopherol and the most effective form is delta-tocotrienol.

Analysis of transcriptional isoforms of collagen types IX, II, and I in the developing avian cornea by competitive polymerase chain reaction.

PubMed

Fitch, J M; Gordon, M K; Gibney, E P; Linsenmayer, T F

1995-01-01

The genes for the alpha 1(IX), alpha 1(II), and alpha 2(I) collagen chains can give rise to different isoforms of mRNA, generated by alternative promotor usage [for alpha 1(IX) and alpha 2(I)] or alternative splicing [for alpha 1(II)]. In this study, we employed competitive reverse transcriptase PCR to quantitate the amounts of transcriptional isoforms for these genes in the embryonic avian cornea from its inception (about 3 1/2 days of development) to 11 days. In order to compare values at different time points, the results were normalized to those obtained for the "housekeeping" enzyme, glycerol-3-phosphate dehydrogenase (G3PDH). These values were compared to those obtained from other tissues (anterior optic cup and cartilage) that synthesize different combinations of the collagen isoforms. We found that, in the cornea, transcripts from the upstream promotor of alpha 1(IX) collagen (termed "long IX") were predominant at stage 18-20 (about 3 1/2 days), but then fell rapidly, and remained at a low level. By 5 days (just before stromal swelling) the major mRNA isoform of alpha 1(IX) was from the downstream promoter (termed "short IX"). The relative amount of transcript for the short form of type IX collagen rose to a peak at about 6 days of development, and then declined. Throughout this period, the predominant transcriptional isoform of the collagen type II gene was IIA (i.e., containing the alternatively spliced exon 2). This indicates that the molecules of type II collagen that are assembled into heterotypic fibrils with type I collagen possess, at least transiently, an amino-terminal globular domain similar to that found in collagen types I, III, and V. For type I, the "bone/tendon" mRNA isoform of the alpha 2(I) collagen gene was predominant; transcripts from the downstream promotor were at basal levels. In other tissues expressing collagen types IX and II, long IX was expressed predominantly with the IIA form in the anterior optic cup at stage 22/23; in 14 1/2 day cartilage, long IX was expressed predominantly along with the IIB form of alpha 1(II). The downstream transcript of the alpha 2(I) gene (Icart) was found at high levels only in cartilage.

Surface expression of heterogeneous nuclear RNA binding protein M4 on Kupffer cell relates to its function as a carcinoembryonic antigen receptor.

PubMed

Bajenova, Olga; Stolper, Eugenia; Gapon, Svetlana; Sundina, Natalia; Zimmer, Regis; Thomas, Peter

2003-11-15

Elevated concentrations of carcinoembryonic antigen (CEA) in the blood are associated with the development of hepatic metastases from colorectal cancers. Clearance of circulating CEA occurs through endocytosis by liver macrophages, Kupffer cells. Previously we identified heterogeneous nuclear ribonucleoproteins M4 (hnRNP M4) as a receptor (CEAR) for CEA. HnRNP M4 has two isoform proteins (p80, p76), the full-length hnRNP M4 (CEARL) and a truncated form (CEARS) with a deletion of 39 amino acids between RNA binding domains 1 and 2, generated by alternative splicing. The present study was undertaken to clarify any isoform-specific differences in terms of their function as CEA receptor and localization. We develop anti-CEAR isoform-specific antibodies and show that both CEAR splicing isoforms are expressed on the surface of Kupffer cells and can function as CEA receptor. Alternatively, in P388D1 macrophages CEARS protein has nuclear and CEARL has cytoplasmic localization. In MIP101 colon cancer and HeLa cells the CEARS protein is localized to the nucleus and CEARL to the cytoplasm. These findings imply that different functions are assigned to CEAR isoforms depending on the cell type. The search of 39 amino acids deleted region against the Prosite data base revealed the presence of N-myristylation signal PGGPGMITIP that may be involved in protein targeting to the plasma membrane. Overall, this report demonstrates that the cellular distribution, level of expression, and relative amount of CEARL and CEARS isoforms determine specificity for CEA binding and the expression of alternative spliced forms of CEAR is regulated in a tissue-specific manner.

The Command and Control Reference Model for Modeling, Simulations, and Technology Applications

DTIC Science & Technology

1994-01-20

NUMBER Basic Research Group (BRG), Technical Panel for C3 (TPC3) Joint Directors of Laboratories (JDL) JDL TPC3 BRG NRaD, San Diego , CA 92152; RADC...wind, ionization, pressure, pollution, ... mesofeature (weather, man-made) cloud, storm, fog, smog, dust, fire, jet stream, smoke, aurora borealis

Bacterial Production, Characterization and Protein Modeling of a Novel Monofuctional Isoform of FAD Synthase in Humans: An Emergency Protein?

PubMed

Leone, Piero; Galluccio, Michele; Barbiroli, Alberto; Eberini, Ivano; Tolomeo, Maria; Vrenna, Flavia; Gianazza, Elisabetta; Iametti, Stefania; Bonomi, Francesco; Indiveri, Cesare; Barile, Maria

2018-01-06

FAD synthase (FADS, EC 2.7.7.2) is the last essential enzyme involved in the pathway of biosynthesis of Flavin cofactors starting from Riboflavin (Rf). Alternative splicing of the human FLAD1 gene generates different isoforms of the enzyme FAD synthase. Besides the well characterized isoform 1 and 2, other FADS isoforms with different catalytic domains have been detected, which are splice variants. We report the characterization of one of these novel isoforms, a 320 amino acid protein, consisting of the sole C-terminal 3'-phosphoadenosine 5'-phosphosulfate (PAPS) reductase domain (named FADS6). This isoform has been previously detected in Riboflavin-Responsive (RR-MADD) and Non-responsive Multiple Acyl-CoA Dehydrogenase Deficiency (MADD) patients with frameshift mutations of FLAD1 gene. To functionally characterize the hFADS6, it has been over-expressed in Escherichia coli and purified with a yield of 25 mg·L -1 of cell culture. The protein has a monomeric form, it binds FAD and is able to catalyze FAD synthesis (k cat about 2.8 min -1 ), as well as FAD pyrophosphorolysis in a strictly Mg 2+ -dependent manner. The synthesis of FAD is inhibited by HgCl₂. The enzyme lacks the ability to hydrolyze FAD. It behaves similarly to PAPS. Combining threading and ab-initio strategy a 3D structural model for such isoform has been built. The relevance to human physio-pathology of this FADS isoform is discussed.

Total pleural covering technique for intractable pneumothorax in patient with Ehlers-Danlos syndrome.

PubMed

Kadota, Yoshihisa; Fukui, Eriko; Kitahara, Naoto; Okura, Eiji; Ohta, Mitsunori

2016-07-01

We report a patient with vascular-type Ehlers-Danlos syndrome (vEDS) who developed pneumothorax and was treated with a total pleural covering technique (TPC). A 24-year-old man developed repeat pneumothorax with intermittent hemo-sputum. Based on unusual radiological manifestations of lung lesions and physical findings, EDS was suspected as an underlying cause of the pneumothorax. Surgical treatment was performed using a mediastinal fat pad and TPC, and no relapse was seen up to 2 years after surgery. TPC is a less invasive surgical approach for selected patients with vEDS. Accurate underlying diagnosis of vEDS and systemic evaluation of vascular complications are necessary before planning surgery.

XAFS of human tyrosine hydroxylase

NASA Astrophysics Data System (ADS)

Meyer, W.; Haavik, J.; Winkler, H.; Trautwein, A. X.; Nolting, H.-F.

1995-02-01

Tyrosine hydroxylase (TH) catalyses the rate-limiting step (hydroxylation of tyrosine to form dihydroxyphenylalanine) in the biosynthetic pathway leading to the catecholamines dopamine, noradrenaline and adrenaline. The human enzyme (hTH) is present in four isoforms, generated by splicing of pre-mRNA. The purified apoenzyme (metal free) binds stoichiometric amounts of iron. The incorporation of Fe(II) results in a rapid and up to 40-fold increase of activity [1]. Besides the coordination of the metal centers in native enzyme we studied the purported inhibition of TH by its immediate products. So we analysed Fe-hTH isoform 1 native as well as oxidized with dopamine and Co-hTH isoform 2.

Effects of deep-fat frying temperature on antioxidant properties of whole wheat doughnuts

USDA-ARS?s Scientific Manuscript database

The total phenolic content (TPC), phenolic acid composition, and in vitro antioxidant capacity of whole wheat donuts fried at 120, 140, 160, or 180'C were determined and compared in two types of wheat to identify the effects of frying temperature. Significant differences (P<0.05) in TPC were observe...

Antioxidant properties of selected fruit cultivars grown in Sri Lanka.

PubMed

Silva, K D R R; Sirasa, M S F

2018-01-01

Extracts of twenty locally available Sri Lankan fruits were analysed for 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, ferrous reducing antioxidant power (FRAP), total phenolic content (TPC), total flavonoid content (TFC) and vitamin C content. The results showed that gooseberry (Phyllanthus emblica 'local') exhibited the highest DPPH scavenging activity (111.25mg ascorbic acid equivalent antioxidant capacity (AEAC)/g), FRAP (1022.05μmol FeSO 4 /g), TPC (915.7mg gallic acid equivalents (GAE)/100g), TFC (873.2mg catechin equivalents (CE)/100g) and vitamin C (136.8mg ascorbic acid equivalents (AAE)/100g), respectively. Sugar apple (Annona squamosa 'local') and star fruit (Averrhoa carambola 'Honey Sweet') obtained the second and third highest antioxidant activities in terms of rankings of FRAP, DPPH activities, TPC, TFC and vitamin C content. Strong correlation between vitamin C, TPC and TFC with FRAP and DPPH showed their contribution to antioxidant capacity. Among the selected fruits, underutilized fruit cultivar gooseberry showed the highest overall antioxidant potential. Copyright © 2016 Elsevier Ltd. All rights reserved.

Fission cross section uncertainties with the NIFFTE TPC

NASA Astrophysics Data System (ADS)

Sangiorgio, Samuele; Niffte Collaboration

2014-09-01

Nuclear data such as neutron-induced fission cross sections play a fundamental role in nuclear energy and defense applications. In recent years, understanding of these systems has become increasingly dependent upon advanced simulation and modeling, where uncertainties in nuclear data propagate in the expected performances of existing and future systems. It is important therefore that uncertainties in nuclear data are minimized and fully understood. For this reason, the Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) uses a Time Projection Chamber (TPC) to measure energy-differential (n,f) cross sections with unprecedented precision. The presentation will discuss how the capabilities of the NIFFTE TPC allow to directly measures systematic uncertainties in fission cross sections, in particular for what concerns fission-fragment identification, and target and beam uniformity. Preliminary results from recent analysis of 238U/235U and 239Pu/235U data collected with the TPC will be presented. This work was performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344.

Optimization of an organic yogurt based on sensorial, nutritional, and functional perspectives.

PubMed

Karnopp, Ariadne Roberto; Oliveira, Katherine Guimarães; de Andrade, Eriel Forville; Postingher, Bruna Mara; Granato, Daniel

2017-10-15

The effects of purple grape juice (PGJ), grape skin flour (GSF), and oligofructose (OLI) on proximate composition, total phenolic content (TPC), antioxidant activity (AA), sensory, physicochemical, and textural properties of yogurts were analyzed using response surface methodology. Multiple regression models were proposed and results showed that PGJ increased the viscosity, AA, and TPC, while GSF increased the ash and total fiber contents of yogurts. GSF and OLI increased the hardness and consistency. A simultaneous optimization was performed to maximize TPC, ash and fibers contents, and sensory acceptance: a yogurt containing 1.7% GSF and 8.0% PGJ had a high fiber (5.60±0.13%) and ash (0.76±0.02%) contents, TPC (28.32±2.10mg GAE/100g), AA toward DPPH (57.85±1.36mg AAE/100g), and total reducing capacity (28.86±5.19mg QE/100g). The optimized yogurt had 79% acceptability index, indicating the use of PGJ and GSF is a feasible alternative to increase the functional properties of yogurts. Copyright © 2017 Elsevier Ltd. All rights reserved.

Temperature Stabilization of the NIFFTE Time Projection Chamber

NASA Astrophysics Data System (ADS)

Hicks, Caleb

2017-09-01

The Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) is a collaboration measuring nuclear fission cross sections for use in advanced nuclear reactors. A neutron beam incident on targets of Uranium-235, Uranium-238, and Plutonium-239 is used to measure the neutron induced fission cross sections for these isotopes. A Time Projection Chamber (TPC) is used to record these reactions. Significant heat is generated by the readout cards mounted on the TPC, which are cooled by fans. One proposed measurement of the experiment is to compare the cross sections of the target to a proton target of gaseous hydrogen. A constant temperature inside the TPC's pressure vessel is desirable to maintain a constant number of hydrogen target atoms. In addition, a constant temperature minimizes the strain and wrinkles on an amplifying mesh inside the TPC. This poster describes the successful work to develop, build, and install a fan controller using a Raspberry Pi, an Arduino, and a custom circuit board to implement an algorithm called Proportional-Integral-Derivative control. This research was supported by US DOE MENP Grant DE-FG02-03ER41243.

DarkSide-20k: A 20 Tonne Two-Phase LAr TPC for Direct Dark Matter Detection at LNGS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Aalseth, C.E.; et al.

Building on the successful experience in operating the DarkSide-50 detector, the DarkSide Collaboration is going to construct DarkSide-20k, a direct WIMP search detector using a two-phase Liquid Argon Time Projection Chamber (LArTPC) with an active (fiducial) mass of 23 t (20 t). The DarkSide-20k LArTPC will be deployed within a shield/veto with a spherical Liquid Scintillator Veto (LSV) inside a cylindrical Water Cherenkov Veto (WCV). Operation of DarkSide-50 demonstrated a major reduction in the dominantmore » $$^{39}$$Ar background when using argon extracted from an underground source, before applying pulse shape analysis. Data from DarkSide-50, in combination with MC simulation and analytical modeling, shows that a rejection factor for discrimination between electron and nuclear recoils of $$\\gt3\\times10^9$$ is achievable. This, along with the use of the veto system, is the key to unlocking the path to large LArTPC detector masses, while maintaining an "instrumental background-free" experiment, an experiment in which less than 0.1 events (other than $$\

Comparison of microwave, ultrasound and accelerated-assisted solvent extraction for recovery of polyphenols from Citrus sinensis peels.

PubMed

Nayak, Balunkeswar; Dahmoune, Farid; Moussi, Kamal; Remini, Hocine; Dairi, Sofiane; Aoun, Omar; Khodir, Madani

2015-11-15

Peel of Citrus sinensis contains significant amounts of bioactive polyphenols that could be used as ingredients for a number of value-added products with health benefits. Extraction of polyphenols from the peels was performed using a microwave-assisted extraction (MAE) technique. The effects of aqueous acetone concentration, microwave power, extraction time and solvent-to-solid ratio on the total phenolic content (TPC), total antioxidant activity (TAA) (using DPPH and ORAC-values) and individual phenolic acids (IPA) were investigated using a response surface method. The TPC, TAA and IPA of peel extracts using MAE was compared with conventional, ultrasound-assisted and accelerated solvent extraction. The maximum predicted TPC under the optimal MAE conditions (51% acetone concentration in water (v/v), 500 W microwave power, 122 s extraction time and 25 mL g(-1) solvent to solid ratio), was 12.20 mg GAE g(-1) DW. The TPC and TAA in MAE extracts were higher than the other three extracts. Copyright © 2015 Elsevier Ltd. All rights reserved.

Regulated expression of a calmodulin isoform alters growth and development in potato.

PubMed

Poovaiah, B W; Takezawa, D; An, G; Han, T J

1996-01-01

A transgene approach was taken to study the consequences of altered expression of a calmodulin isoform on plant growth and development. Eight genomic clones of potato calmodulin (PCM1 to 8) have been isolated and characterized (Takezawa et al., 1995). Among the potato calmodulin isoforms studied, PCM1 differs from the other isoforms because of its unique amino acid substitutions. Transgenic potato plants were produced carrying sense construct of PCM1 fused to the CaMV 35S promoter. Transgenic plants showing a moderate increase in PCM1 mRNA exhibited strong apical dominance, produced elongated tubers, and were taller than the controls. Interestingly, the plants expressing the highest level of PCM1 mRNA did not form underground tubers. Instead, these transgenic plants produced aerial tubers when allowed to grow for longer periods. The expression of different calmodulin isoforms (PCM1, 5, 6, and 8) was studied in transgenic plants. Among the four potato calmodulin isoforms, only the expression of PCM1 mRNA was altered in transgenic plants, while the expression of other isoforms was not significantly altered. Western analysis revealed increased PCM1 protein in transgenic plants, indicating that the expression of both mRNA and protein are altered in transgenic plants. These results suggest that increasing the expression of PCM1 alters growth and development in potato plants.

Differential expression of Na+, K(+)-ATPase α-1 isoforms during seawater acclimation in the amphidromous galaxiid fish Galaxias maculatus.

PubMed

Urbina, Mauricio A; Schulte, Patricia M; Bystriansky, Jason S; Glover, Chris N

2013-04-01

Inanga (Galaxias maculatus) is an amphidromous fish with a well-known capacity to withstand a wide range of environmental salinities. To investigate the molecular mechanisms facilitating acclimation of inanga to seawater, several isoforms of the Na(+), K(+)-ATPase ion transporter were identified. This included three α-1 (a, b and c), an α-2 and two α-3 (a and b) isoforms. Phylogenetic analysis showed that the inanga α-1a and α-1b formed a clade with the α-1a and α-1b isoforms of rainbow trout, while another clade contained the α-1c isoforms of these species. The expression of all the α-1 isoforms was modulated after seawater exposure (28‰). In gills, the expression of the α-1a isoform was progressively down-regulated after seawater exposure, while the expression of the α-1b isoform was up-regulated. The α-1c isoform behaved similarly to the α-1a, although changes were less dramatic. Physiological indicators of salinity acclimation matched the time frame of the changes observed at the molecular level. A 24-h osmotic shock period was highlighted by small increases in plasma osmolality, plasma Na(+) and a decrease in muscle tissue water content. Thereafter, these values returned close to their pre-exposure (freshwater) values. Na(+), K(+)-ATPase activity showed a decreasing trend over the first 72 h following seawater exposure, but activity increased after 240 h. Our results indicate that inanga is an excellent osmoregulator, an ability that is conferred by the rapid activation of physiological and molecular responses to salinity change.

Antimicrobial and antioxidant activities of Cichorium intybus root extract using orthogonal matrix design.

PubMed

Liu, Haitao; Wang, Quanzhen; Liu, Yuyan; Chen, Guo; Cui, Jian

2013-02-01

Solvent, impregnation time, sonication repetitions, and ultrasonic power were important factors in the process of ultrasound-assisted extraction from chicory (Cichorium intybus) root, while there were no studies about optimizing these 4 factors for extract yield, total phenolic content (TPC), antioxidant, antibacterial, and antifungal activity of the extracts using orthogonal matrix design. The present research demonstrated that the solvent composition played a significant role in the improving extract yield, TPC, antioxidant, and antibacterial activities. The other 3 factors had inequable effect on different purposes, ultrasonic power could improve TPC and antioxidant activity, but long time of extraction lowered antioxidant activity. The TPC increased from 22.34 to 27.87 mg GAE (gallic acid equivalents)/100 g (dry extracts) with increasing solvent polarity. The half inhibition concentration (IC(50,) μg/mL) of the radical scavenging activity of the chicory extracts ranged from 281.00 to 983.33 μg/mL. The content of caffeoylquinic acids of root extract, which was extracted by the optimal combination was 0.104%. Several extracts displayed antibacterial activities against Escherichia coli, Staphylococcus aureus, Bacillus thuringiensis, Bacillus subtilis, and Salmonella typhi, while Penicillium sp. and Aspergillus sp. resisted against all the extracts. Combination of 70% ethanol v/v, 24-h impregnation time, 3 sonication rounds, and 300-W ultrasonic input power was found to be the optimal combination for the chicory extract yield, TPC, antioxidant activity, and antibacterial activity. © 2012 Institute of Food Technologists®

Effect of soaking, boiling, and steaming on total phenolic contentand antioxidant activities of cool season food legumes.

PubMed

Xu, Baojun; Chang, Sam K C

2008-09-01

The effects of soaking, boiling and steaming processes on the total phenolic components and antioxidant activity in commonly consumed cool season food legumes (CSFL's), including green pea, yellow pea, chickpea and lentil were investigated. As compared to original unprocessed legumes, all processing steps caused significant (p<0.05) decreases in total phenolic content (TPC), DPPH free radical scavenging activity (DPPH) in all tested CSFL's. All soaking and atmospheric boiling treatments caused significant (p<0.05) decreases in oxygen radical absorbing capacity (ORAC). However, pressure boiling and pressure steaming caused significant (p<0.05) increases in ORAC values. Steaming treatments resulted in a greater retention of TPC, DPPH, and ORAC values in all tested CSFL's as compared to boiling treatments. To obtain cooked legumes with similar palatability and firmness, pressure boiling shortened processing time as compared to atmospheric boiling, resulted in insignificant differences in TPC, DPPH for green and yellow pea. However, TPC and DPPH in cooked lentils differed significantly between atmospheric and pressure boiling. As compared to atmospheric processes, pressure processes significantly increased ORAC values in both boiled and steamed CSFL's. Greater TPC, DPPH and ORAC values were detected in boiling water than that in soaking and steaming water. Boiling also caused more solid loss than steaming. Steam processing exhibited several advantages in retaining the integrity of the legume appearance and texture of the cooked product, shortening process time, and greater retention of antioxidant components and activities. Copyright © 2008 Elsevier Ltd. All rights reserved.

Supramolecular architectures in Co(II) and Cu(II) complexes with thiophene-2-carboxylate and 2-amino-4,6-dimethoxypyrimidine ligands.

PubMed

Karthikeyan, Ammasai; Thomas Muthiah, Packianathan; Perdih, Franc

2016-05-01

The coordination chemistry of mixed-ligand complexes continues to be an active area of research since these compounds have a wide range of applications. Many coordination polymers and metal-organic framworks are emerging as novel functional materials. Aminopyrimidine and its derivatives are flexible ligands with versatile binding and coordination modes which have been proven to be useful in the construction of organic-inorganic hybrid materials and coordination polymers. Thiophenecarboxylic acid, its derivatives and their complexes exhibit pharmacological properties. Cobalt(II) and copper(II) complexes of thiophenecarboxylate have many biological applications, for example, as antifungal and antitumor agents. Two new cobalt(II) and copper(II) complexes incorporating thiophene-2-carboxylate (2-TPC) and 2-amino-4,6-dimethoxypyrimidine (OMP) ligands have been synthesized and characterized by X-ray diffraction studies, namely (2-amino-4,6-dimethoxypyrimidine-κN)aquachlorido(thiophene-2-carboxylato-κO)cobalt(II) monohydrate, [Co(C5H3O2S)Cl(C6H9N3O2)(H2O)]·H2O, (I), and catena-poly[copper(II)-tetrakis(μ-thiophene-2-carboxylato-κ(2)O:O')-copper(II)-(μ-2-amino-4,6-dimethoxypyrimidine-κ(2)N(1):N(3))], [Cu2(C5H3O2S)4(C6H9N3O2)]n, (II). In (I), the Co(II) ion has a distorted tetrahedral coordination environment involving one O atom from a monodentate 2-TPC ligand, one N atom from an OMP ligand, one chloride ligand and one O atom of a water molecule. An additional water molecule is present in the asymmetric unit. The amino group of the coordinated OMP molecule and the coordinated carboxylate O atom of the 2-TPC ligand form an interligand N-H...O hydrogen bond, generating an S(6) ring motif. The pyrimidine molecules also form a base pair [R2(2)(8) motif] via a pair of N-H...N hydrogen bonds. These interactions, together with O-H...O and O-H...Cl hydrogen bonds and π-π stacking interactions, generate a three-dimensional supramolecular architecture. The one-dimensional coordination polymer (II) contains the classical paddle-wheel [Cu2(CH3COO)4(H2O)2] unit, where each carboxylate group of four 2-TPC ligands bridges two square-pyramidally coordinated Cu(II) ions and the apically coordinated OMP ligands bridge the dinuclear copper units. Each dinuclear copper unit has a crystallographic inversion centre, whereas the bridging OMP ligand has crystallographic twofold symmetry. The one-dimensional polymeric chains self-assemble via N-H...O, π-π and C-H...π interactions, generating a three-dimensional supramolecular architecture.

Tree age, fruit size and storage conditions affect levels of ascorbic acid, total phenolic concentrations and total antioxidant activity of 'Kinnow' mandarin juice.

PubMed

Khalid, Samina; Malik, Aman U; Khan, Ahmad S; Shahid, Muhammad; Shafique, Muhammad

2016-03-15

Bioactive compounds (ascorbic acid, total phenolics and total antioxidants) are important constituents of citrus fruit juice; however, information with regard to their concentrations and changes in relation to tree age and storage conditions is limited. 'Kinnow' (Citrus nobilis Lour × Citrus deliciosa Tenora) mandarin juice from fruit of three tree ages (6, 18 and 35 years old) and fruit sizes (large, medium and small) were examined for their bioactive compounds during 7 days under ambient storage conditions (20 ± 2 °C and 60-65% relative humidity (RH)) and during 60 days under cold storage (4 ± 1 °C and 75-80% RH) conditions. Under ambient conditions, a reduction in total phenolic concentrations (TPC) and in total antioxidant activity (TAA) was found for the juice from all tree ages and fruit sizes. Overall, fruit from 18-year-old trees had higher mean TPC (95.86 µg mL(-1) ) and TAA (93.68 mg L(-1) ), as compared to 6 and 35-year-old trees. Likewise, in cold storage, TAA decreased in all fruit size groups from 18 and 35-year-old trees. In all tree age and fruit size groups, TPC decreased initially during 15 days of cold storage and then increased gradually with increase in storage duration. Ascorbic acid concentrations showed an increasing trend in all fruit size groups from 35-year-old trees. Overall, during cold storage, fruit from 18-year-old trees maintained higher mean ascorbic acid (33.05 mg 100 mL(-1) ) concentrations, whereas fruit from 6-year-old trees had higher TAA (153.1 mg L(-1) ) and TPC (115.1 µg mL(-1) ). Large-sized fruit had higher ascorbic acid (32.08 mg 100 mL(-1) ) concentrations and TAA (157.5 mg L(-1) ). Fruit from 18-year-old trees maintained higher TPC and TAA under ambient storage conditions, whereas fruit from 6-year-old trees maintained higher TPC and TAA during cold storage. Small-sized fruit had higher TPC after ambient temperature storage, whereas large fruit size showed higher ascorbic acid concentrations and TAA after cold storage. © 2015 Society of Chemical Industry.

Effect of heat treatment on the antioxidant activity of extracts from citrus peels.

PubMed

Jeong, Seok-Moon; Kim, So-Young; Kim, Dong-Ryul; Jo, Seong-Chun; Nam, K C; Ahn, D U; Lee, Seung-Cheol

2004-06-02

The effect of heat treatment on the antioxidant activity of extracts from Citrus unshiu peels was evaluated. Citrus peels (CP) (5 g) were placed in Pyrex Petri dishes (8.0 cm diameter) and heat-treated at 50, 100, or 150 degrees C for 10, 20, 30, 40, 50, and 60 min in an electric muffle furnace. After heat treatment, 70% ethanol extract (EE) and water extract (WE) (0.1 g/10 mL) of CP were prepared, and total phenol contents (TPC), radical scavenging activity (RSA), and reducing power of the extracts were determined. The antioxidant activities of CP extracts increased as heating temperature increased. For example, heat treatment of CP at 150 degrees C for 60 min increased the TPC, RSA, and reducing power of EE from 71.8 to 171.0 microM, from 29.64 to 64.25%, and from 0.45 to 0.82, respectively, compared to non-heat-treated control. In the case of WE from CP heat-treated at the same conditions (150 degrees C for 60 min), the TPC, RSA, and reducing power also increased from 84.4 to 204.9 microM, from 15.81 to 58.26%, and from 0.27 to 0.96, respectively. Several low molecular weight phenolic compounds such as 2,3-diacetyl-1-phenylnaphthalene, ferulic acid, p-hydroxybenzaldoxime, 5-hydroxyvaleric acid, 2,3-diacetyl-1-phenylnaphthalene, and vanillic acid were newly formed in the CP heated at 150 degrees C for 30 min. These results indicated that the antioxidant activity of CP extracts was significantly affected by heating temperature and duration of treatment on CP and that the heating process can be used as a tool for increasing the antioxidant activity of CP.

Microbial effect of steam vacuum pasteurisation implemented after slaughtering and dressing of sheep and lamb.

PubMed

Hassan, Ammar Ali; Skjerve, Eystein; Bergh, Claus; Nesbakken, Truls

2015-01-01

The main objective of the study was to assess the effect of steam vacuum pasteurisation on carcass contamination with focus on Escherichia coli, Enterobacteriaceae and total plate count (TPC). Additionally, the effect of an additional tryptone soy agar (TSA) step for resuscitation of Enterobacteriaceae after steam vacuum pasteurisation was investigated. Steam vacuum pasteurisation was applied at a temperature of >82°C for a duration of 10s on sheep and lamb carcasses (n=120). Samples were taken immediately: i) after trimming just before the use of steam vacuum and ii) after use of steam vacuum. Nordic Committee on Food Analysis methods were used in microbial analyses. The differences in log reduction were found significant for all of the three microorganisms (p<0.05). For TPC, the general reduction was a 0.65 log10 in the number of colony forming units (CFU) per cm(2). For E. coli, the median reduction effect on carcasses positive before decontamination was 1.1 log10 CFU/cm(2). A large variability of the effect was however found, with 50% of the figures ranging from a 0.24 to 1.62 log10 CFU/cm(2) reduction and a 10-90% range of 0-2.1. The number of positive carcasses with Enterobacteriaceae after steam vacuum pasteurisation was higher in samples where TSA+violet red bile glucose agar (VRBGA) was used compared to samples where only VRBGA was used (p<0.01). Steam vacuum pasteurisation was found efficient in reducing the total count, read as TPC, as well as the level of E. coli and Enterobacteriaceae. Copyright © 2014 Elsevier Ltd. All rights reserved.

Multiple Isoforms of ANRIL in Melanoma Cells: Structural Complexity Suggests Variations in Processing.

PubMed

Sarkar, Debina; Oghabian, Ali; Bodiyabadu, Pasani K; Joseph, Wayne R; Leung, Euphemia Y; Finlay, Graeme J; Baguley, Bruce C; Askarian-Amiri, Marjan E

2017-06-27

The long non-coding RNA ANRIL , antisense to the CDKN2B locus, is transcribed from a gene that encompasses multiple disease-associated polymorphisms. Despite the identification of multiple isoforms of ANRIL , expression of certain transcripts has been found to be tissue-specific and the characterisation of ANRIL transcripts remains incomplete. Several functions have been associated with ANRIL . In our judgement, studies on ANRIL functionality are premature pending a more complete appreciation of the profusion of isoforms. We found differential expression of ANRIL exons, which indicates that multiple isoforms exist in melanoma cells. In addition to linear isoforms, we identified circular forms of ANRIL ( circANRIL ). Further characterisation of circANR IL in two patient-derived metastatic melanoma cell lines (NZM7 and NZM37) revealed the existence of a rich assortment of circular isoforms. Moreover, in the two melanoma cell lines investigated, the complements of circANRIL isoforms were almost completely different. Novel exons were also discovered. We also found the family of linear ANRIL was enriched in the nucleus, whilst the circular isoforms were enriched in the cytoplasm and they differed markedly in stability. With respect to the variable processing of circANRIL species, bioinformatic analysis indicated that intronic Arthrobacter luteus (Alu) restriction endonuclease inverted repeats and exon skipping were not involved in selection of back-spliced exon junctions. Based on our findings, we hypothesise that " ANRIL " has wholly distinct dual sets of functions in melanoma. This reveals the dynamic nature of the locus and constitutes a basis for investigating the functions of ANRIL in melanoma.

Post-translational modification and stability of low molecular weight cyclin E.

PubMed

Mull, B B; Cox, J; Bui, T; Keyomarsi, K

2009-09-03

Our laboratory has previously described the presence of five tumor-specific low molecular weight isoforms of cyclin E in both tumor cell lines and breast cancer patient biopsies. We have also shown that one of these low forms arises from an alternate start site, whereas the other four appear as two sets of doublets following cleavage through an elastase-like enzyme. However, the origin of both sets of doublets was unknown. Here, we demonstrate that the larger isoform of each doublet is the result of phosphorylation at a key degradation site. Through site-directed mutagenesis of different phosphorylation sites within the cyclin E protein, we discovered that phosphorylation of threonine 395 is responsible for generating the larger isoform of each doublet. Because phosphorylation of threonine 395 has been linked to the proteasome-mediated degradation of full length cyclin E, we examined the stability of T395A phospho-mutants in both non-tumorigenic mammary epithelial cells and tumor cells. The results revealed that the low molecular weight isoforms appear to be stable in both a tumor cell line and a non-tumor forming cell line regardless of the presence of this critical phosphorylation site. The stability of low molecular weight cyclin E may have implications for both tumorigenesis and treatment of tumors expressing them.

Characterization of HSP90 isoforms in transformed bovine leukocytes infected with Theileria annulata

PubMed Central

Kinnaird, Jane H.; Singh, Meetali; Gillan, Victoria; Weir, William; Calder, Ewen D. D.; Hostettler, Isabel; Shiels, Brian R.

2016-01-01

Summary HSP90 chaperones are essential regulators of cellular function, as they ensure the appropriate conformation of multiple key client proteins. Four HSP90 isoforms were identified in the protozoan parasite Theileria annulata. Partial characterization was undertaken for three and localization confirmed for cytoplasmic (TA12105), endoplasmic reticulum (TA06470), and apicoplast (TA10720) forms. ATPase activity and binding to the HSP90 inhibitor geldanamycin were demonstrated for recombinant TA12105, and all three native forms could be isolated to varying extents by binding to geldanamycin beads. Because it is essential, HSP90 is considered a potential therapeutic drug target. Resistance to the only specific Theileriacidal drug is increasing, and one challenge for design of drugs that target the parasite is to limit the effect on the host. An in vitro cell culture system that allows comparison between uninfected bovine cells and the T. annulata‐infected counterpart was utilized to test the effects of geldanamycin and the derivative 17‐AAG. T. annulata‐infected cells had greater tolerance to geldanamycin than uninfected cells yet exhibited significantly more sensitivity to 17‐AAG. These findings suggest that parasite HSP90 isoform(s) can alter the drug sensitivity of infected host cells and that members of the Theileria HSP90 family are potential targets worthy of further investigation. PMID:27649068

Structure-activity relationships in defensin dimers: a novel link between beta-defensin tertiary structure and antimicrobial activity.

PubMed

Campopiano, Dominic J; Clarke, David J; Polfer, Nick C; Barran, Perdita E; Langley, Ross J; Govan, John R W; Maxwell, Alison; Dorin, Julia R

2004-11-19

Defensins are cationic antimicrobial peptides that have a characteristic six-cysteine motif and are important components of the innate immune system. We recently described a beta-defensin-related peptide (Defr1) that had potent antimicrobial activity despite having only five cysteines. Here we report a relationship between the structure and activity of Defr1 through a comparative study with its six cysteine-containing analogue (Defr1 Y5C). Against a panel of pathogens, we found that oxidized Defr1 had significantly higher activity than its reduced form and the oxidized and reduced forms of Defr1 Y5C. Furthermore, Defr1 displayed activity against Pseudomonas aeruginosa in the presence of 150 mm NaCl, whereas Defr1 Y5C was inactive. By using nondenaturing gel electrophoresis and Fourier transform ion cyclotron resonance mass spectrometry, we observed Defr1 and Defr1 Y5C dimers. Two complementary fragmentation techniques (collision-induced dissociation and electron capture dissociation) revealed that Defr1 Y5C dimers form by noncovalent, weak association of monomers that contain three intramolecular disulfide bonds. In contrast, Defr1 dimers are resistant to collision-induced dissociation and are only dissociated into monomers by reduction using electron capture. This is indicative of Defr1 dimerization being mediated by an intermolecular disulfide bond. Proteolysis and peptide mass mapping revealed that Defr1 Y5C monomers have beta-defensin disulfide bond connectivity, whereas oxidized Defr1 is a complex mixture of dimeric isoforms with as yet unknown inter- and intramolecular connectivities. Each isoform contains one intermolecular and four intramolecular disulfide bonds, but because we were unable to resolve the isoforms by reverse phase chromatography, we could not assign each isoform with a specific antimicrobial activity. We conclude that the enhanced activity and stability of this mixture of Defr1 dimeric isoforms are due to the presence of an intermolecular disulfide bond. This first description of a covalently cross-linked member of the defensin family provides further evidence that the antimicrobial activity of a defensin is linked to its ability to form stable higher order structures.

The Technical Communicator as Advocate: Integrating a Social Justice Approach in Technical Communication

ERIC Educational Resources Information Center

Jones, Natasha N.

2016-01-01

This article argues for the need for a social justice approach to technical communication research and pedagogy. Given previous calls by scholars in technical and professional communication (TPC) for an attention to diversity, inclusion, and equality, the author examines the place and purpose of social justice in TPC and provides useful approaches…

Phrenic Long-Term Facilitation Requires PKCθ Activity within Phrenic Motor Neurons

PubMed Central

Devinney, Michael J.; Fields, Daryl P.; Huxtable, Adrianne G.; Peterson, Timothy J.; Dale, Erica A.

2015-01-01

Acute intermittent hypoxia (AIH) induces a form of spinal motor plasticity known as phrenic long-term facilitation (pLTF); pLTF is a prolonged increase in phrenic motor output after AIH has ended. In anesthetized rats, we demonstrate that pLTF requires activity of the novel PKC isoform, PKCθ, and that the relevant PKCθ is within phrenic motor neurons. Whereas spinal PKCθ inhibitors block pLTF, inhibitors targeting other PKC isoforms do not. PKCθ is highly expressed in phrenic motor neurons, and PKCθ knockdown with intrapleural siRNAs abolishes pLTF. Intrapleural siRNAs targeting PKCζ, an atypical PKC isoform expressed in phrenic motor neurons that underlies a distinct form of phrenic motor plasticity, does not affect pLTF. Thus, PKCθ plays a critical role in spinal AIH-induced respiratory motor plasticity, and the relevant PKCθ is localized within phrenic motor neurons. Intrapleural siRNA delivery has considerable potential as a therapeutic tool to selectively manipulate plasticity in vital respiratory motor neurons. PMID:26019328

Isoform-Specific Upregulation of Palladin in Human and Murine Pancreas Tumors

PubMed Central

Goicoechea, Silvia M.; Bednarski, Brian; Stack, Christianna; Cowan, David W.; Volmar, Keith; Thorne, Leigh; Cukierman, Edna; Rustgi, Anil K.; Brentnall, Teresa; Hwang, Rosa F.; McCulloch, Christopher A. G.; Yeh, Jen Jen; Bentrem, David J.; Hochwald, Steven N.; Hingorani, Sunil R.

2010-01-01

Pancreatic ductal adenocarcinoma (PDA) is a lethal disease with a characteristic pattern of early metastasis, which is driving a search for biomarkers that can be used to detect the cancer at an early stage. Recently, the actin-associated protein palladin was identified as a candidate biomarker when it was shown that palladin is mutated in a rare inherited form of PDA, and overexpressed in many sporadic pancreas tumors and premalignant precursors. In this study, we analyzed the expression of palladin isoforms in murine and human PDA and explored palladin's potential use in diagnosing PDA. We performed immunohistochemistry and immunoblot analyses on patient samples and tumor-derived cells using an isoform-selective monoclonal antibody and a pan-palladin polyclonal antibody. Immunoblot and real-time quantitative reverse transcription-PCR were used to quantify palladin mRNA levels in human samples. We show that there are two major palladin isoforms expressed in pancreas: 65 and 85–90 kDa. The 65 kDa isoform is expressed in both normal and neoplastic ductal epithelial cells. The 85–90 kDa palladin isoform is highly overexpressed in tumor-associated fibroblasts (TAFs) in both primary and metastatic tumors compared to normal pancreas, in samples obtained from either human patients or genetically engineered mice. In tumor-derived cultured cells, expression of palladin isoforms follows cell-type specific patterns, with the 85–90 kDa isoform in TAFs, and the 65 kDa isoform predominating in normal and neoplastic epithelial cells. These results suggest that upregulation of 85–90 kDa palladin isoform may play a role in the establishment of the TAF phenotype, and thus in the formation of a desmoplastic tumor microenvironment. Thus, palladin may have a potential use in the early diagnosis of PDA and may have much broader significance in understanding metastatic behavior. PMID:20436683

Isolation of complementary DNA clones encoding pathogenesis-related proteins P and Q, two acidic chitinases from tobacco.

PubMed Central

Payne, G; Ahl, P; Moyer, M; Harper, A; Beck, J; Meins, F; Ryals, J

1990-01-01

Complementary DNA clones encoding two isoforms of the acidic endochitinase (chitinase, EC 3.2.1.14) from tobacco were isolated. Comparison of amino acid sequences deduced from the cDNA clones and the sequence of peptides derived from purified proteins show that these clones encode the pathogenesis-related proteins PR-P and PR-Q. The cDNA inserts were not homologous to either the bacterial form of chitinase or the form from cucumber but shared significant homology to the basic form of chitinase from tobacco and bean. The acidic isoforms of tobacco chitinase did not contain the amino-terminal, cysteine-rich "hevein" domain found in the basic isoforms, indicating that this domain, which binds chitin, is not essential for chitinolytic activity. The accumulation of mRNA for the pathogenesis-related proteins PR-1, PR-R, PR-P, and PR-Q in Xanthi.nc tobacco leaves following infection with tobacco mosaic virus was measured by primer extension. The results indicate that the induction of these proteins during the local necrotic lesion response to the virus is coordinated at the mRNA level. Images PMID:2296608

Preliminary X-ray crystallographic studies of a tetrameric phospholipase A{sub 2} formed by two isoforms of crotoxin B from Crotalus durissus terrificus venom

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marchi-Salvador, D. P.; Corrêa, L. C.; Salvador, G. H. M.

2007-12-01

Crotoxin B is a basic phospholipase A{sub 2} found in the venom of C. durissus terrificus and is one of the subunits that constitute crotoxin. Here, the crystallization, X-ray diffraction data collection and molecular-replacement solution of a novel tetrameric complex formed by two dimers of crotoxin B isoforms are presented. Crotoxin B is a basic phospholipase A{sub 2} found in the venom of Crotalus durissus terrificus and is one of the subunits that constitute crotoxin. This heterodimeric toxin, which is the main component of C. d. terrificus venom, is completed by an acidic, nontoxic and non-enzymatic component (crotoxin A) andmore » is involved in important envenomation effects, such as neurological disorders, myotoxicity and renal failure. Although crotoxin was first crystallized in 1938, no crystal structure is currently available for crotoxin, crotoxin A or crotoxin B. In this work, the crystallization, X-ray diffraction data collection to 2.28 Å resolution and molecular-replacement solution of a novel tetrameric complex formed by two dimers of crotoxin B isoforms (CB1 and CB2) is presented.« less

Effects of different isoforms of apoE on aggregation of the α-synuclein protein implicated in Parkinson's disease.

PubMed

Emamzadeh, Fatemeh Nouri; Aojula, Harmesh; McHugh, Patrick C; Allsop, David

2016-04-08

Parkinson's disease is a progressive brain disorder due to the degeneration of dopaminergic neurons in the substantia nigra. The accumulation of aggregated forms of α-synuclein protein into Lewy bodies is one of the characteristic features of this disease although the pathological role of any such protein deposits in causing neurodegeneration remains elusive. Here, the effects of different apolipoprotein E isoforms (apoE2, apoE3, apoE4) on the aggregation of α-synuclein in vitro were examined using thioflavin T assays and also an immunoassay to detect the formation of multimeric forms. Our results revealed that the aggregation of α-synuclein is influenced by apoE concentration. At low concentrations of apoE (<15nM), all of the isoforms were able to increase the aggregation of α-synuclein (50μM), with apoE4 showing the greatest stimulatory effect. This is in contrast to a higher concentration (>15nM) of these isoforms, where a decrease in the aggregation of α-synuclein was noted. The data show that exceptionally low levels of apoE may seed α-syn aggregation, which could potentially lead to the pathogenesis of α-synuclein-induced neurodegeneration. On the other hand, higher levels of apoE could potentially lower the degree of α-synuclein aggregation and confer protection. The differential effects noted with apoE4 could explain why this particular isoform results in an earlier age of onset for Parkinson's disease. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Physicochemical, nutritional, and sensory qualities of wine grape pomace fortified baked goods.

PubMed

Walker, Rebecca; Tseng, Angela; Cavender, George; Ross, Andrew; Zhao, Yanyun

2014-09-01

Wine grape pomace (WGP) as a source of antioxidant dietary fiber (DF) was used to fortify baked goods, including breads, muffins, and brownies. Pinot Noir WGP (RWGP) and Pinot Grigio WGP (WWGP) substituted wheat flour at concentration of 5%, 10%, and 15% for bread, 10%, 15%, 20%, and 25% RWGP for brownies, and 5%, 10%, and 15% RWGP or 10%, 15%, and 20% WWGP for muffins. The finished products were evaluated for total phenolic content (TPC), radical scavenging activity (RSA), and total DF, as well as physicochemical and sensory properties. WGP flour blends were also tested for solvent retention capacity (SRC). The highest TPC and RSA values for bread and muffins were achieved in 15% RWGP fortified samples with TPC and RSA values of 68.32 mg gallic acid equivalent (GAE)/serving and 80.70 AAE mg/serving, respectively for bread, and 2164 mg GAE/serving and 1526 mg AAE/serving, respectively for muffins. Brownies fortified with 10% RWGP had the highest RSA value (115.52 mg AAE/serving) while the control had the highest TPC value (1152 mg GAE/serving). Breads and muffins with 15% RWGP and brownies with 25% RWGP had the highest amount of DF (6.33, 12.32, and 7.73 g/serving, respectively). Sensory evaluation concluded that there is no difference in overall liking of 5% and 10% RWGP breads and muffins or 15% and 20% WGP brownies compared to the controls. This study demonstrated that WGP is a viable functional ingredient in bakery goods to increase TPC, RSA, and DF in consumer's diets. © 2014 Institute of Food Technologists®

Phenolic Content and Antioxidant Activity during the Development of 'Brookfield' and 'Mishima' Apples.

PubMed

Stanger, Mayara C; Steffens, Cristiano A; Soethe, Cristina; Moreira, Marcelo A; do Amarante, Cassandro V T

2017-05-03

The aim of this study was to characterize the changes in the contents of total (TPC) and individual (IPC) phenolic compounds, the total antioxidant activity (TAA) in the peel and pulp, and total anthocyanins (TAN) in the peel during the development of the fruits of 'Brookfield' and 'Mishima' apple trees. 'Brookfield' apples were harvested from the 49th to the 138th days after full bloom (DAFB) and 'Mishima' apples from the 45th to the 172th DAFB. In the pulp, the IPC, TPC, and TAA rapidly reduced at 75 and 79 DAFB for the 'Brookfield' and 'Mishima' apples, respectively, and then remained constant until commercial maturity. In the peel of 'Brookfield' apples there was a reduction in the TPC and TAA at 79 DAFB. The quercetin 3-galactoside, epicatechin, and procyanidin B2 contents reduced up to 107 DAFB with a subsequent increase in the values at commercial maturity. In the peel of 'Mishima' apples there was a reduction in the TPC, TAA, epicatechin, and procyanidin B1 and B2 contents at 130 DAFB, with a subsequent increase until commercial maturity. The TAN content in the peel increased during the 2 and 4 weeks prior to commercial maturity for 'Brookfield' and 'Mishima' apples, respectively. In the pulp and peel of both cultivars there was a reduction in the IPC, TPC, and TAA as the development proceeded. On nearing commercial maturity, there was an increase in the contents of quercetin 3-galactoside, epicatechin, procyanidin B2, and TAN in the peel for both cultivars.

Structure of Voltage-gated Two-pore Channel TPC1 from Arabidopsis thaliana

PubMed Central

Guo, Jiangtao; Zeng, Weizhong; Chen, Qingfeng; Lee, Changkeun; Chen, Liping; Yang, Yi; Cang, Chunlei; Ren, Dejian; Jiang, Youxing

2015-01-01

Two-pore channels (TPCs) contain two copies of a Shaker-like six-transmembrane (6-TM) domain in each subunit and are ubiquitously expressed in both animals and plants as organellar cation channels. Here, we present the first crystal structure of a vacuolar two-pore channel from Arabidopsis thaliana, AtTPC1, which functions as a homodimer. AtTPC1 activation requires both voltage and cytosolic Ca2+. Ca2+ binding to the cytosolic EF-hand domain triggers conformational changes coupled to the pair of pore-lining inner helices (IS6 helices) from the first 6-TM domains, whereas membrane potential only activates the second voltage-sensing domain (VSD2) whose conformational changes are coupled to the pair of inner helices (IIS6 helices) from the second 6-TM domains. Luminal Ca2+ or Ba2+ can modulate voltage activation by stabilizing VSD2 in the resting state and shifts voltage activation towards more positive potentials. Our Ba2+ bound AtTPC1 structure reveals a voltage sensor in the resting state, providing hitherto unseen structural insight into the general voltage-gating mechanism among voltage-gated channels. PMID:26689363

Solvent-Free Microwave-Assisted Extraction of Polyphenols from Olive Tree Leaves: Antioxidant and Antimicrobial Properties.

PubMed

Şahin, Selin; Samli, Ruya; Tan, Ayşe Seher Birteksöz; Barba, Francisco J; Chemat, Farid; Cravotto, Giancarlo; Lorenzo, José M

2017-06-24

Response surface methodology (RSM) and artificial neural networks (ANN) were evaluated and compared in order to decide which method was the most appropriate to predict and optimize total phenolic content (TPC) and oleuropein yields in olive tree leaf ( Olea europaea ) extracts, obtained after solvent-free microwave-assisted extraction (SFMAE). The SFMAE processing conditions were: microwave irradiation power 250-350 W, extraction time 2-3 min, and the amount of sample 5-10 g. Furthermore, the antioxidant and antimicrobial activities of the olive leaf extracts, obtained under optimal extraction conditions, were assessed by several in vitro assays. ANN had better prediction performance for TPC and oleuropein yields compared to RSM. The optimum extraction conditions to recover both TPC and oleuropein were: irradiation power 250 W, extraction time 2 min, and amount of sample 5 g, independent of the method used for prediction. Under these conditions, the maximal yield of oleuropein (0.060 ± 0.012 ppm) was obtained and the amount of TPC was 2.480 ± 0.060 ppm. Moreover, olive leaf extracts obtained under optimum SFMAE conditions showed antibacterial activity against S. aureus and S. epidermidis , with a minimum inhibitory concentration (MIC) value of 1.25 mg/mL.

Multi-elemental composition and antioxidant properties of strawberry tree (Arbutus unedo L.) honey from the coastal region of Croatia: Risk-benefit analysis.

PubMed

Tariba Lovaković, Blanka; Lazarus, Maja; Brčić Karačonji, Irena; Jurica, Karlo; Živković Semren, Tanja; Lušić, Dražen; Brajenović, Nataša; Pelaić, Zdenka; Pizent, Alica

2018-01-01

The concentration of 23 major and trace elements, total phenolic content (TPC) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were determined in nine samples of strawberry tree honey and compared to other types of unifloral honeys. The most abundant elements in strawberry tree honey were potassium, calcium, magnesium and sodium, ranging between 1276 and 2367, 95.2-154, 14.4-74.4 and 13.4-64.3mg/kg, respectively. Strawberry tree honey had generally higher TPC (range: 0.314-0.522g GA/kg) and DPPH (1.94-4.45mM TE/kg) compared to other analysed unifloral honeys. A strong positive relationship was found between TPC and DPPH, TPC and concentration of homogentisic acid (HGA), chemical marker of strawberry tree honey, and between DPPH and HGA. Regarding daily intake of essential elements, strawberry tree honey can be considered nutritionally richer than the majority of unifloral honeys available in Croatia, while contribution to tolerable intake set for potentially toxic elements was very low, corresponding to pristine areas. Copyright © 2017 Elsevier GmbH. All rights reserved.

An image-guided radiotherapy decision support framework incorporating a Bayesian network and visualization tool.

PubMed

Hargrave, Catriona; Deegan, Timothy; Bednarz, Tomasz; Poulsen, Michael; Harden, Fiona; Mengersen, Kerrie

2018-05-17

To describe a Bayesian network (BN) and complementary visualization tool that aim to support decision-making during online cone-beam computed tomography (CBCT)-based image-guided radiotherapy (IGRT) for prostate cancer patients. The BN was created to represent relationships between observed prostate, proximal seminal vesicle (PSV), bladder and rectum volume variations, an image feature alignment score (FAS TV _ OAR ), delivered dose, and treatment plan compliance (TPC). Variables influencing tumor volume (TV) targeting accuracy such as intrafraction motion, and contouring and couch shift errors were also represented. A score of overall TPC (FAS global ) and factors such as image quality were used to inform the BN output node providing advice about proceeding with treatment. The BN was quantified using conditional probabilities generated from published studies, FAS TV _ OAR /global modeling, and a survey of IGRT decision-making practices. A new IGRT visualization tool (IGRT REV ), in the form of Mollweide projection plots, was developed to provide a global summary of residual errors after online CBCT-planning CT registration. Sensitivity and scenario analyses were undertaken to evaluate the performance of the BN and the relative influence of the network variables on TPC and the decision to proceed with treatment. The IGRT REV plots were evaluated in conjunction with the BN scenario testing, using additional test data generated from retrospective CBCT-planning CT soft-tissue registrations for 13/36 patients whose data were used in the FAS TV _ OAR /global modeling. Modeling of the TV targeting errors resulted in a very low probability of corrected distances between the CBCT and planning CT prostate or PSV volumes being within their thresholds. Strength of influence evaluation with and without the BN TV targeting error nodes indicated that rectum- and bladder-related network variables had the highest relative importance. When the TV targeting error nodes were excluded from the BN, TPC was sensitive to observed PSV and rectum variations while the decision to treat was sensitive to observed prostate and PSV variations. When root nodes were set so the PSV and rectum variations exceeded thresholds, the probability of low TPC increased to 40%. Prostate and PSV variations exceeding thresholds increased the likelihood of repositioning or repeating patient preparation to 43%. Scenario testing using the test data from 13 patients, demonstrated two cases where the BN provided increased high TPC probabilities, despite some of the prostate and PSV volume variation metrics not being within tolerance. The IGRT REV tool was effective in highlighting and quantifying where TV and OAR variations occurred, supporting the BN recommendation to reposition the patient or repeat their bladder and bowel preparation. In another case, the IGRT REV tool was also effective in highlighting where PSV volume variation significantly exceeded tolerance when the BN had indicated to proceed with treatment. This study has demonstrated that both the BN and IGRT REV plots are effective tools for inclusion in a decision support system for online CBCT-based IGRT for prostate cancer patients. Alternate approaches to modeling TV targeting errors need to be explored as well as extension of the BN to support offline IGRT decisions related to adaptive radiotherapy. © 2018 American Association of Physicists in Medicine.

Development of intestinal ion-transporting mechanisms during smoltification and seawater acclimation in Atlantic salmon Salmo salar

USGS Publications Warehouse

Sundh, Henrik; Nilsen, Tom O.; Lindström, Jenny; Hasselberg-Frank, Linda; Stefansson, Sigurd O.; McCormick, Stephen D.; Sundell, K.

2014-01-01

This study investigated the expression of ion transporters involved in intestinal fluid absorption and presents evidence for developmental changes in abundance and tissue distribution of these transporters during smoltification and seawater (SW) acclimation of Atlantic salmonSalmo salar. Emphasis was placed on Na+, K+-ATPase (NKA) and Na+, K+, Cl− co-transporter (NKCC) isoforms, at both transcriptional and protein levels, together with transcription of chloride channel genes. The nka α1c was the dominant isoform at the transcript level in both proximal and distal intestines; also, it was the most abundant isoform expressed in the basolateral membrane of enterocytes in the proximal intestine. This isoform was also abundantly expressed in the distal intestine in the lower part of the mucosal folds. The protein expression of intestinal Nkaα1c increased during smoltification. Immunostaining was localized to the basal membrane of the enterocytes in freshwater (FW) fish, and re-distributed to a lateral position after SW entry. Two other Nka isoforms, α1a and α1b, were expressed in the intestine but were not regulated to the same extent during smoltification and subsequent SW transfer. Their localization in the intestinal wall indicates a house-keeping function in excitatory tissues. The absorptive form of the NKCC-like isoform (sub-apically located NKCC2 and/or Na+, Cl−co-transporter) increased during smoltification and further after SW transfer. The cellular distribution changed from a diffuse expression in the sub-apical regions during smoltification to clustering of the transporters closer to the apical membrane after entry to SW. Furthermore, transcript abundance indicates that the mechanisms necessary for exit of chloride ions across the basolateral membrane and into the lateral intercellular space are present in the form of one or more of three different chloride channels: cystic fibrosis transmembrane conductance regulator I and II and chloride channel 3.

Phenolic acids, anthocyanins, and antioxidant capacity in rice (Oryza sativa L.) grains at four stages of development after flowering.

PubMed

Shao, Yafang; Xu, Feifei; Sun, Xiao; Bao, Jinsong; Beta, Trust

2014-01-15

This study investigated differences in total phenolic content (TPC), antioxidant capacity, and phenolic acids in free, conjugated and bound fractions of white (unpolished), red and black rice at 1-, 2-, and 3-weeks of grain development after flowering and at maturity. Unlike the TPC (mg/100g) of white rice (14.6-33.4) and red rice (66.8-422.2) which was significantly higher at 1-week than at later stages, the TPC of black rice (56.5-82.0) was highest at maturity. The antioxidant capacity measured by DPPH radical scavenging and ORAC methods generally followed a similar trend as TPC. Only black rice had detectable anthocyanins (26.5-174.7mg/100g). Cyanidin-3-glucoside (C3G) and peonidin-3-glucoside (P3G) were the main anthocyanins in black rice showing significantly higher levels at 2- and 3-weeks than at 1-week development and at maturity. At all stages, the phenolic acids existed mainly in the bound form as detected by HPLC and confirmed by LC-MS/MS. Black rice (20.1-31.7mg/100g) had higher total bound phenolic acids than white rice and red rice (7.0-11.8mg/100g). Protocatechuic acid was detected in red rice and black rice with relatively high levels at 1-week development (1.41mg/100g) and at maturity (4.48mg/100g), respectively. Vanillic acid (2.4-5.4mg/100g) was detected only in black rice where it peaked at maturity. p-Coumaric acid (<3.5mg/100g) did not differ significantly at most stages with somewhat high levels at 1-week for red and black rice. Ferulic acid (4.0-17.9mg/100g), the most abundant bound phenolic acid, had an inconsistent trend with higher levels being observed in black rice where it peaked at maturity. Isoferulic acid levels (0.8-1.6mg/100g) were generally low with slightly elevated values being observed at maturity. Overall black rice had higher total bound phenolic acids than white and red rice while white rice at all stages of development after flowering. Copyright © 2013 Elsevier Ltd. All rights reserved.

TPC Proteins Are Phosphoinositide-activated Sodium-selective Ion Channels in Endosomes and Lysosomes

PubMed Central

Wang, Xiang; Zhang, Xiaoli; Dong, Xian-ping; Samie, Mohammad; Li, Xinran; Cheng, Xiping; Goschka, Andrew; Shen, Dongbiao; Zhou, Yandong; Harlow, Janice; Zhu, Michael X.; Clapham, David E.; Ren, Dejian; Xu, Haoxing

2012-01-01

Summary Mammalian Two-Pore Channels (TPC1, 2; TPCN1, TPCN2) encode ion channels in intracellular endosomes and lysosomes and were proposed to mediate endolysosomal calcium release triggered by the second messenger, nicotinic acid adenine dinucleotide phosphate (NAADP). By directly recording TPCs in endolysosomes from wild-type and TPC double knockout mice, here we show that, in contrast to previous conclusions, TPCs are in fact sodium-selective channels activated by PI(3,5)P2, and are not activated by NAADP. Moreover, the primary endolysosomal ion is Na+, not K+, as had been previously assumed. These findings suggest that the organellar membrane potential may undergo large regulatory changes, and may explain the specificity of PI(3,5)P2 in regulating the fusogenic potential of intracellular organelles. PMID:23063126

Development of the GEM-TPC X-ray Polarimeter with the Scalable Readout System

NASA Astrophysics Data System (ADS)

Kitaguchi, Takao; Hayato, Asami; Iwakiri, Wataru; Takeuchi, Yoko; Kubota, Megu; Nishida, Kazuki; Enoto, Teruaki; Tamagawa, Toru

2018-02-01

We have developed a gaseous Time Projection Chamber (TPC) containing a single-layered foil of a gas electron multiplier (GEM) to open up a new window on cosmic X-ray polarimetry in the 2-10 keV band. The micro-pattern TPC polarimeter in combination with the Scalable Readout System produced by the RD51 collaboration has been built as an engineering model to optimize detector parameters and improve polarimeter sensitivity. The polarimeter was characterized with unpolarized X-rays from an X-ray generator in a laboratory and polarized X-rays on the BL32B2 beamline at the SPring-8 synchrotron radiation facility. Preliminary results show that the polarimeter has a comparable modulation factor to a prototype of the flight one.

Performance of 20:1 multiplexer for large area charge readouts in directional dark matter TPC detectors

NASA Astrophysics Data System (ADS)

Ezeribe, A. C.; Robinson, M.; Robinson, N.; Scarff, A.; Spooner, N. J. C.; Yuriev, L.

2018-02-01

More target mass is required in current TPC based directional dark matter detectors for improved detector sensitivity. This can be achieved by scaling up the detector volumes, but this results in the need for more analogue signal channels. A possible solution to reducing the overall cost of the charge readout electronics is to multiplex the signal readout channels. Here, we present a multiplexer system in expanded mode based on LMH6574 chips produced by Texas Instruments, originally designed for video processing. The setup has a capability of reducing the number of readouts in such TPC detectors by a factor of 20. Results indicate that the important charge distribution asymmetry along an ionization track is retained after multiplexed signals are demultiplexed.

Study of Electromagnetic Interactions with the MicroBooNE Detector

NASA Astrophysics Data System (ADS)

Caratelli, David; MicroBooNE Collaboration

2017-01-01

MicroBooNE is an experiment which employs the Liquid Argon Time Projection Chamber (LArTPC) detector technology to study neutrinos produced with the Fermilab Booster Neutrino Beam. As for any accelerator-based detector interested in studying neutrino oscillations, it is essential to be able to identify and reconstruct the kinematic properties of electrons and photons produced in μν and νe interactions. We report current progress in reconstructing electron and photon electromagnetic (EM) showers using data from the MicroBooNE LArTPC. These studies cover EM showers in the tens to hundreds of MeV energy range; they lay the foundation for MicroBooNE's investigation of the excess of low-energy EM events reported by MiniBooNE, and are of interest to the wider LArTPC neutrino community.

Poring over two-pore channel pore mutants

PubMed Central

Penny, Christopher J.; Patel, Sandip

2016-01-01

Two-pore channels are members of the voltage-gated ion channel superfamily. They localise to the endolysosomal system and are likely targets for the Ca2+ mobilising messenger NAADP. In this brief review, we relate mutagenesis of the TPC pore to a recently published homology model and discuss how pore mutants are informing us of TPC function. Molecular physiology of these ubiquitous proteins is thus emerging. PMID:27226934

The LArIAT Experiment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nutini, Irene

2017-09-20

A short overview of the Liquid Argon In A Testbeam (LArIAT) experiment hosted at Fermilab is reported. This program supports the Liquid Argon Time Projection Chamber (LArTPC) Neutrino Experiments at Fermilab. The LArIAT program consists of a calibration of a LArTPC in a dedicated charged particle beamline. The first total pion interaction cross section measurement ever made on argon is presented here (preliminary result).

Research and Development Toward Massive Liquid Argon Time Projection Chambers for Neutrino Detection

DOE Office of Scientific and Technical Information (OSTI.GOV)

Thiesse, Matthew

Liquid argon (LAr) time projection chambers (TPC) have rapidly increased in importance as particle detectors throughout the past four decades. While much research has been completed, there are still many areas which require further development to build and operate the next generation LAr TPC experiment, such as the Deep Underground Neutrino Experiment (DUNE). These include high voltage breakdown, argon purification and purity monitoring, and vacuum ultraviolet (VUV) scintillation light measurement. Visual monitoring of high voltage breakdown is helpful in allowing assessment of the performance of high voltage component design. Thus, a system of cryogenic cameras, the first of its kind,more » was developed for use in a large LAr cryostat, without the need for additional electronics heating. The system functioned without problem for 50 days at cryogenic temperature, with some degradation of image quality, and provided a useful monitor for the DUNE 35-ton cryogenics systems. The system did not observe any high voltage breakdowns during the run. Further development of the concept is ongoing for future installation in other experiments. The monitoring of LAr purity using TPC data is a fundamental study for LAr TPC experiments. However, the study has not been performed for a large LAr TPC in the presence of high electronic noise. Custom software was developed and validated for the accurate reconstruction of signals in noisy TPC data. The results of the reconstruction were used to successfully measure the LAr electron lifetime with an uncertainty comparable to alternate methods of measurement. The electron lifetime of the 35-ton Phase II run is determined to be 4.12 ± 0.17 (stat.) ±0.40 (syst.) ms. For general purpose research and development of high purity LAr as a particle detection medium, a dedicated test stand was designed, constructed, and commissioned. The system is used to test the gaseous photomultiplier (GPM) performance at cryogenic temperatures. The GPM functions with photoelectron multiplication at 77 K, at a reduced gain. Further study is required to show the detector’s direct sensitivity to LAr VUV scintillation light.« less

A large ungated TPC with GEM amplification

NASA Astrophysics Data System (ADS)

Berger, M.; Ball, M.; Fabbietti, L.; Ketzer, B.; Arora, R.; Beck, R.; Böhmer, F. V.; Chen, J.-C.; Cusanno, F.; Dørheim, S.; García, F.; Hehner, J.; Herrmann, N.; Höppner, C.; Kaiser, D.; Kis̆, M.; Kleipa, V.; Konorov, I.; Kunkel, J.; Kurz, N.; Leifels, Y.; Müllner, P.; Münzer, R.; Neubert, S.; Rauch, J.; Schmidt, C. J.; Schmitz, R.; Soyk, D.; Vandenbroucke, M.; Voss, B.; Walther, D.; Zmeskal, J.

2017-10-01

A Time Projection Chamber (TPC) is an ideal device for the detection of charged particle tracks in a large volume covering a solid angle of almost 4 π. The high density of hits on a given particle track facilitates the task of pattern recognition in a high-occupancy environment and in addition provides particle identification by measuring the specific energy loss for each track. For these reasons, TPCs with Multiwire Proportional Chamber (MWPC) amplification have been and are widely used in experiments recording heavy-ion collisions. A significant drawback, however, is the large dead time of the order of 1 ms per event generated by the use of a gating grid, which is mandatory to prevent ions created in the amplification region from drifting back into the drift volume, where they would severely distort the drift path of subsequent tracks. For experiments with higher event rates this concept of a conventional TPC operating with a triggered gating grid can therefore not be applied without a significant loss of data. A continuous readout of the signals is the more appropriate way of operation. This, however, constitutes a change of paradigm with considerable challenges to be met concerning the amplification region, the design and bandwidth of the readout electronics, and the data handling. A mandatory prerequisite for such an operation is a sufficiently good suppression of the ion backflow from the avalanche region, which otherwise limits the tracking and particle identification capabilities of such a detector. Gas Electron Multipliers (GEM) are a promising candidate to combine excellent spatial resolution with an intrinsic suppression of ions. In this paper we describe the design, construction and the commissioning of a large TPC with GEM amplification and without gating grid (GEM-TPC). The design requirements have driven innovations in the construction of a light-weight field-cage, a supporting media flange, the GEM amplification and the readout system, which are presented in this paper. We further describe the support infrastructure such as gas, cooling and slow control. Finally, we report on the operation of the GEM-TPC in the FOPI experiment, and describe the calibration procedures which are applied to achieve the design performance of the device.

Characterization of bioactive compounds of Annona cherimola L. leaves using a combined approach based on HPLC-ESI-TOF-MS and NMR.

PubMed

Díaz-de-Cerio, Elixabet; Aguilera-Saez, Luis Manuel; Gómez-Caravaca, Ana María; Verardo, Vito; Fernández-Gutiérrez, Alberto; Fernández, Ignacio; Arráez-Román, David

2018-06-01

Annona cherimola Mill. (cherimoya) has widely been used as food crop. The leaves of this tree possess several health benefits, which are, in general, attributed mainly to its bioactive composition. However, literature concerning a comprehensive characterization based on a combined approach, which consists of nuclear magnetic resonance (NMR) and high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF-MS), from these leaves is scarce. Thus, the aim of this work was to study the polar profile of full extracts of cherimoya leaves by using these tools. Thus, a total of 77 compounds have been characterized, 12 of which were identified by both techniques. Briefly, 23 compounds were classified as amino acids, organic acids, carbohydrates, cholines, phenolic acid derivatives, and flavonoids by NMR, while 66 metabolites were divided into sugars, amino acids, phenolic acids and derivatives, flavonoids, phenylpropanoids, and other polar compounds by HPLC-TOF-MS. It is worth mentioning that different solvent mixtures were tested and the total phenolic content in the extracts quantified (TPC via HPLC-TOF-MS). The tendency observed was EtOH/water 80/20 (v/v) (17.0 ± 0.2 mg TPC/g leaf dry weight (d.w.)) ≥ acetone/water 70/30 (v/v) (16.1 ± 0.7 mg TPC/g leaf d.w.) > EtOH/water 70/30 (v/v) (14.0 ± 0.3 mg TPC/g leaf d.w.) > acetone/water 80/20 (v/v) (13.5 ± 0.4 mg TPC/g leaf d.w.). Importantly, flavonoids derivatives were between 63 and 76% of the TPC in those extracts. Major compounds were sucrose, glucose (α and β), and proline, and chlorogenic acid and rutin for NMR and HPLC-TOF-MS, respectively. Graphical abstract The combined use of LC-HRMS and NMR is a potential synergic combination for a comprehensive metabolite composition of cherimoya leaves.

C-terminal motifs in promyelocytic leukemia protein isoforms critically regulate PML nuclear body formation.

PubMed

Li, Chuang; Peng, Qiongfang; Wan, Xiao; Sun, Haili; Tang, Jun

2017-10-15

Promyelocytic leukemia protein (PML) nuclear bodies (NBs), which are sub-nuclear protein structures, are involved in a variety of important cellular functions. PML-NBs are assembled by PML isoforms, and contact between small ubiquitin-like modifiers (SUMOs) with the SUMO interaction motif (SIM) are critically involved in this process. PML isoforms contain a common N-terminal region and a variable C-terminus. However, the contribution of the C-terminal regions to PML-NB formation remains poorly defined. Here, using high-resolution microscopy, we show that mutation of the SIM distinctively influences the structure of NBs formed by each individual PML isoform, with that of PML-III and PML-V minimally changed, and PML-I and PML-IV dramatically impaired. We further identify several C-terminal elements that are important in regulating NB structure and provide strong evidence to suggest that the 8b element in PML-IV possesses a strong ability to interact with SUMO-1 and SUMO-2, and critically participates in NB formation. Our findings highlight the importance of PML C-termini in NB assembly and function, and provide molecular insight into the PML-NB assembly of each distinctive isoform. © 2017. Published by The Company of Biologists Ltd.

Plectin isoforms as organizers of intermediate filament cytoarchitecture

PubMed Central

Winter, Lilli

2011-01-01

Intermediate filaments (IFs) form cytoplamic and nuclear networks that provide cells with mechanical strength. Perturbation of this structural support causes cell and tissue fragility and accounts for a number of human genetic diseases. In recent years, important additional roles, nonmechanical in nature, were ascribed to IFs, including regulation of signaling pathways that control survival and growth of the cells, and vectorial processes such as protein targeting in polarized cellular settings. The cytolinker protein plectin anchors IF networks to junctional complexes, the nuclear envelope and cytoplasmic organelles and it mediates their cross talk with the actin and tubulin cytoskeleton. These functions empower plectin to wield significant influence over IF network cytoarchitecture. Moreover, the unusual diversity of plectin isoforms with different N termini and a common IF-binding (C-terminal) domain enables these isoforms to specifically associate with and thereby bridge IF networks to distinct cellular structures. Here we review the evidence for IF cytoarchitecture being controlled by specific plectin isoforms in different cell systems, including fibroblasts, endothelial cells, lens fibers, lymphocytes, myocytes, keratinocytes, neurons and astrocytes, and discuss what impact the absence of these isoforms has on IF cytoarchitecture-dependent cellular functions. PMID:21866256

Linear Streptomyces plasmids form superhelical circles through interactions between their terminal proteins

PubMed Central

Tsai, Hsiu-Hui; Huang, Chih-Hung; Tessmer, Ingrid; Erie, Dorothy A.; Chen, Carton W.

2011-01-01

Linear chromosomes and linear plasmids of Streptomyces possess covalently bound terminal proteins (TPs) at the 5′ ends of their telomeres. These TPs are proposed to act as primers for DNA synthesis that patches the single-stranded gaps at the 3′ ends during replication. Most (‘archetypal’) Streptomyces TPs (designated Tpg) are highly conserved in size and sequence. In addition, there are a number of atypical TPs with heterologous sequences and sizes, one of which is Tpc that caps SCP1 plasmid of Streptomyces coelicolor. Interactions between the TPs on the linear Streptomyces replicons have been suggested by electrophoretic behaviors of TP-capped DNA and circular genetic maps of Streptomyces chromosomes. Using chemical cross-linking, we demonstrated intramolecular and intermolecular interactions in vivo between Tpgs, between Tpcs and between Tpg and Tpc. Interactions between the chromosomal and plasmid telomeres were also detected in vivo. The intramolecular telomere interactions produced negative superhelicity in the linear DNA, which was relaxed by topoisomerase I. Such intramolecular association between the TPs poses a post-replicational complication in the formation of a pseudo-dimeric structure that requires resolution by exchanging TPs or DNA. PMID:21109537

Feasibility study and sensory test of turmeric tamarind traditional drink with various processing

NASA Astrophysics Data System (ADS)

Rudyatmi, E.; Bintari, S. H.; Iswari, R. S.

2018-03-01

Turmeric tamarind traditional drink as anti-oxidant, good for health, and has a potency to developed. The objectives of this study are to determine the feasibility and the favorite level of consumer towards two turmerics tamarind traditional drink which processed differently. Feasibility depended on sensory test and microbe's contamination according to a quality standard of BPOM and complemented with an anti-bacterial test. The anti-bacterial test is determined by a diameter of inhibition zone towards E coli and B subtilist. Consumer’s favorites level is determined by organoleptic test to 40 testers. Sensory test results to form, odor, taste, and color are normal. TPC first traditional drink 6,9 x 102 col/gr and TKK ≤1,0x10 col/gr, TPC second traditional drink 2,0 x 101 col/gr and TKK ≤1,0x10 col/gr; all tests toward E coli, Salmonella SP, Staphylococcus aureus, Pseudomonas aeruginosa, shigella sp negative/gr; inhibiting capabilities towards B.subtilis and E coli of first traditional drink > second traditional drink. Inhibiting capabilities towards B.subtilis > E coli. All parameters comply with BPOM standard. Most of the tester love the first method turmeric tamarind traditional drink.

Expression of two isoforms of CD44 in human endometrium.

PubMed

Behzad, F; Seif, M W; Campbell, S; Aplin, J D

1994-10-01

The distribution of the cell-surface adhesion glycoprotein CD44 in human endometrium was examined by immunofluorescence using six monoclonal antibodies to epitopes common to all forms of the molecule, and by reverse transcription-polymerase chain reaction (RT-PCR). Immunoreactivity was observed throughout the menstrual cycle in stroma, vessels, glandular, and luminal epithelium. Variations in staining intensity were observed, especially in the epithelial compartment. CD44 was also expressed strongly by decidualized stromal cells of first-trimester pregnancy. No systematic variation of immunoreactivity was observed with stages of the normal cycle, but a fraction (25%) of the specimens lacked reactivity in the epithelium. To determine the molecular size of the epithelial isoform, an immunoprecipitation technique was developed using surface-radioiodinated, detergent-extracted glands. This indicated the presence at the cell surface of a single dominant CD44E species with an approximate molecular mass of 130 kDa. RT-PCR was used to investigate the isoforms present in whole endometrial tissue, isolated gland fragments, and Ishikawa endometrial carcinoma cells. Complementary DNA produced from total endometrial mRNA was PCR-amplified across the splice junction between exons 5 and 15. Transcripts corresponding to the hyaluronate receptor CD44H as well as a larger isoform were identified. CD44H was absent, or very scarce, in cDNA from purified gland epithelium. In contrast, Ishikawa cells expressed this form abundantly. The glands and Ishikawa cells also expressed CD44E containing sequences encoded by exons 12, 13, and 14. These data demonstrate the presence of CD44 in human endometrium and decidua, and show that different isoforms of CD44 are associated with tissue compartments in which different functional roles can be anticipated.

Sex change strategy and the aromatase genes.

PubMed

Gardner, L; Anderson, T; Place, A R; Dixon, B; Elizur, A

2005-04-01

Sequential hermaphroditism is a common reproductive strategy in many teleosts. Steroid production is known to mediate both the natural and induced sex change, yet beyond this the physiology directing this process has received little attention. Cytochrome P450 aromatase is a key enzyme in the hormonal pathway catalysing the conversion of sex steroids, androgens to oestrogens, and thus is highly relevant to the process of sex change. This study reports the isolation of cDNA sequences for aromatase isoforms CYP19A1 and CYP19A2 from teleost species representing three forms of sexual hermaphroditism: Lates calcarifer (protandry), Cromileptes altivelis (protogyny), and Gobiodon histrio (bi-directional). Deduced amino acid analysis of these isoforms with other reported isoforms from gonochoristic (single sex) teleosts revealed 56-95% identity within the same isoform while only 48-65% identity between isoforms irrespective of species and sexual strategy. Phylogenetic analysis supported this result separating sequences into isoform exclusive clades in spite of species apparent evolutionary distance. Furthermore, this study isolates 5' flanking regions of all above genes and describes putative cis-acting elements therein. Elements identified include steroidogenic factor 1 binding site (SF-1), oestrogen response element (ERE), progesterone response element (PRE), androgen response element (ARE), glucocorticoid response elements (GRE), peroxisome proliferator-activated receptor alpha/retinoid X receptor alpha heterodimer responsive element (PPARalpha/RXRalpha), nuclear factor kappabeta (NF-kappabeta), SOX 5, SOX 9, and Wilms tumor suppressor (WTI). A hypothetical in vivo model was constructed for both isoforms highlighting potential roles of these putative cis-acting elements with reference to normal function and sexual hermaphroditism.

Liquid argon scintillation detection utilizing wavelength-shifting plates and light guides

NASA Astrophysics Data System (ADS)

Howard, B.

2018-02-01

In DUNE, the event timing provided by the detection of the relatively prompt scintillation photons will improve spatial resolution in the drift direction of the time-projection chamber (TPC) and is especially useful for non-beam physics topics such as supernova neutrinos and nucleon decay. The baseline design for the first 10kt single phase TPC fits the photon detector system in the natural gap between the wire planes of adjacent TPC volumes. A prototype photon detector design utilizes wavelength-shifter coated plates to convert the vacuum ultraviolet scintillation light to the optical and commercially-produced wavelength-shifting light guides to trap some of this light and transport it to an array of silicon photomultipliers at the end. This system and the testing performed to characterize the system and determine the efficiency are discussed.

Liquid Argon Scintillation Detection Utilizing Wavelength-Shifting Plates and Light Guides

DOE Office of Scientific and Technical Information (OSTI.GOV)

Howard, B.

In DUNE, the event timing provided by the detection of the relatively prompt scintillation photons will improve spatial resolution in the drift direction of the time-projection chamber (TPC) and is especially useful for non-beam physics topics such as supernova neutrinos and nucleon decay. The baseline design for the first 10kt single phase TPC fits the photon detector system in the natural gap between the wire planes of adjacent TPC volumes. A prototype photon detector design utilizes wavelength-shifter coated plates to convert the vacuum ultraviolet scintillation light to the optical and commercially-produced wavelength-shifting light guides to trap some of this lightmore » and transport it to an array of silicon photomultipliers at the end. This system and the testing performed to characterize the system and determine the efficiency are discussed.« less

Commissioning of the Active-Target Time Projection Chamber

NASA Astrophysics Data System (ADS)

Bradt, J.; Bazin, D.; Abu-Nimeh, F.; Ahn, T.; Ayyad, Y.; Beceiro Novo, S.; Carpenter, L.; Cortesi, M.; Kuchera, M. P.; Lynch, W. G.; Mittig, W.; Rost, S.; Watwood, N.; Yurkon, J.

2017-12-01

The Active-Target Time Projection Chamber (AT-TPC) was recently built and commissioned at the National Superconducting Cyclotron Laboratory at Michigan State University. This gas-filled detector uses an active-target design where the gas acts as both the tracking medium and the reaction target. Operating inside a 2T solenoidal magnetic field, the AT-TPC records charged particle tracks that can be reconstructed to very good energy and angular resolutions. The near- 4 π solid angle coverage and thick target of the detector are well-suited to experiments with low secondary beam intensities. In this paper, the design and instrumentation of theAT-TPC are described along with the methods used to analyze the data it produces. A simulation of the detector's performance and some results from its commissioning with a radioactive 46Ar beam are also presented.

Combined Effect of Random Transmit Power Control and Inter-Path Interference Cancellation on DS-CDMA Packet Mobile Communications

NASA Astrophysics Data System (ADS)

Kudoh, Eisuke; Ito, Haruki; Wang, Zhisen; Adachi, Fumiyuki

In mobile communication systems, high speed packet data services are demanded. In the high speed data transmission, throughput degrades severely due to severe inter-path interference (IPI). Recently, we proposed a random transmit power control (TPC) to increase the uplink throughput of DS-CDMA packet mobile communications. In this paper, we apply IPI cancellation in addition to the random TPC. We derive the numerical expression of the received signal-to-interference plus noise power ratio (SINR) and introduce IPI cancellation factor. We also derive the numerical expression of system throughput when IPI is cancelled ideally to compare with the Monte Carlo numerically evaluated system throughput. Then we evaluate, by Monte-Carlo numerical computation method, the combined effect of random TPC and IPI cancellation on the uplink throughput of DS-CDMA packet mobile communications.

MICROMEGAS calibration for ACTAR TPC

NASA Astrophysics Data System (ADS)

Mauss, B.; Roger, T.; Pancin, J.; Damoy, S.; Grinyer, G. F.

2018-02-01

Active targets, such as the ACtive TARget and Time Projection Chamber (ACTAR TPC) being developed at GANIL, are detection systems that operate on the basis of a time projection chamber but where the filling gas also serves as a thick target for nuclear reactions. In nuclear physics experiments, the energy resolution is of primary importance to identify the reaction products and to precisely reconstruct level schemes of nuclei. These measurements are based on the energy deposited on a pixelated pad plane. A MICROMEGAS detector is used in ACTAR TPC for the ionization electron collection and amplification, and it is a major contributor to the energy dispersion through, for example, inhomogeneities of the amplification gap. A variation of one percent in the gap can lead to an amplitude variation of more than two percent which is of the same order as the resolution obtained with an energy deposition of 5 MeV. One way to calibrate the pad plane is through the use of a two dimensional source scanning table. It is used to calibrate the gain inhomogeneities and, using MAGBOLTZ calculations, deduce the corresponding gap variations. The inverse of this method would allow the relative gain variations to be calculated for the different gas mixtures and pressures used in experiments with ACTAR TPC.

Exclusive Muon-Neutrino Charged Current Muon Plus Any Number of Protons Topologies In ArgoNeuT

DOE Office of Scientific and Technical Information (OSTI.GOV)

Partyka, Kinga Anna

2013-01-01

Neutrinos remain among the least understood fundamental particles even after decades of study. As we enter the precision era o f neutrino measurements bigger and more sophisticated detectors have emerged. The leading candidate among them is a Liquid Argon Time Projection Chamber (LArTPC ) detector technology due to its bubble-like chamber imaging, superb background rejection and scalability. I t is a perfect candidate that w ill aim to answer the remaining questions of the nature o f neutrino and perhaps our existence. Studying neutrinos with a detector that employs detection via beautiful images o f neutrino interactions can be bothmore » illuminating and surprising. The analysis presented here takes the full advantage of the LArTPC power by exploiting the first topological analysis of charged current muon neutrino p + N p , muon and any number of protons, interactions with the ArgoNeuT LArTPC experiment on an argon target. The results presented here are the first that address the proton multiplicity at the vertex and the proton kinematics. This study also addresses the importance o f nuclear effects in neutrino interactions. Furthermore, the developed here reconstruction techniques present a significant step forward for this technology and can be employed in the future LArTPC detectors.« less

15-Deoxy-Δ12,14-prostaglandin J2 Induces Apoptosis and Upregulates SOCS3 in Human Thyroid Cancer Cells

PubMed Central

Trindade-da-Silva, Carlos Antônio; Reis, Carolina Fernandes; Vecchi, Lara; Napimoga, Marcelo Henrique; Sperandio, Marcelo; Matias Colombo, Bruna França; Alves, Patrícia Terra; Ward, Laura Sterian; Ueira-Vieira, Carlos; Goulart, Luiz Ricardo

2016-01-01

The cyclopentenone prostaglandin 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) is a natural ligand of peroxisome proliferator-activated receptor gamma (PPAR-γ) and a potential mediator of apoptosis in cancer cells. In the present study, we evaluated the effect of 15d-PGJ2 in human thyroid papillary carcinoma cells (TPC-1) using different doses of 15d-PGJ2 (0.6 to 20 μM) to determine IC50 (9.3 μM) via the MTT assay. The supernatant culture medium of the TPC-1 cells that was treated either with 15d-PGJ2 or with vehicle (control) for 24 hours was assessed for IL-6 secretion via CBA assay. RT-qPCR was used to evaluate mRNA expression of IL-6, SOCS1, SOCS3, and STAT3. TPC-1 cells treated with 15d-PGJ2 decreased the secretion and expression of IL-6 and STAT3, while it increased SOCS1 and SOCS3. Overall, we demonstrated that 15d-PGJ2 downregulated IL-6 signaling pathway and led TPC-1 cells into apoptosis. In conclusion, 15d-PGJ2 shows the potential to become a new therapeutic approach for thyroid tumors. PMID:27190500

Simulations of Charged-Current Supernova νe Events in a Liquid Argon Time Projection Chamber

NASA Astrophysics Data System (ADS)

Gardiner, Steven; Grant, Christopher; Pantic, Emilija; Svoboda, Robert

2016-03-01

Although it is still in its infancy, the study of supernova neutrinos has proven to be a fertile topic for fundamental science. A mere two dozen events recorded from supernova 1987A, the only supernova neutrino source observed so far, have led to numerous publications on a wide variety of topics. This bountiful scientific harvest has prompted the neutrino physics community to prepare to make more detailed observations of the neutrinos that will be produced in the next nearby supernova. Because of their unique νe sensitivity, liquid argon time projection chamber (LArTPC) experiments such as DUNE (Deep Underground Neutrino Experiment) have the potential to make valuable contributions to this detection effort. To better understand the expected SN νe signal in a LArTPC, we have developed a Monte Carlo event generator called MARLEY (Model of Argon Reaction Low-Energy Yields) for charged-current νe reactions on argon. By combining MARLEY with LArSoft, a LArTPC simulation package, we have obtained the most detailed predictions currently available for the response of a LArTPC to supernova νe. We will discuss the implications of these results for the design and operation of LArTPCs sensitive to SN neutrinos.

High-performance size-exclusion chromatography studies on the formation and distribution of polar compounds in camellia seed oil during heating.

PubMed

Feng, Hong-Xia; Sam, Rokayya; Jiang, Lian-Zhou; Li, Yang; Cao, Wen-Ming

Camellia seed oil (CSO) is rich in oleic acid and has a high number of active components, which give the oil high nutritional value and a variety of biological activity. The aim of the present study was to determine the changes in the content and distribution of total polar compounds (TPC) in CSO during heating. TPC were isolated by means of preparative flash chromatography and further analyzed by high-performance size-exclusion chromatography (HPSEC). The TPC content of CSO increased from 4.74% to 25.29%, showing a significantly lower formation rate as compared to that of extra virgin olive oil (EVOO) and soybean oil (SBO) during heating. Furthermore, heating also resulted in significant differences (P<0.05) in the distribution of TPC among these oils. Though the content of oxidized triacylglycerol dimers, oxidized triacylglycerol oligomers, and oxidized triacylglycerol monomers significantly increased in all these oils, their increased percentages were much less in CSO than those in EVOO, indicating that CSO has a greater ability to resist oxidation. This work may be useful for the food oil industry and consumers in helping to choose the correct oil and to decide on the useful lifetime of the oil.

MICROWAVE-ASSISTED EXTRACTION OF PHENOLIC COMPOUNDS FROM POLYGONUM MULTIFLORUM THUNB. ROOTS.

PubMed

Quoc, Le Pham Tan; Muoi, Nguyen Van

2016-01-01

The aim of this study was to determine the best extraction conditions for total phenolic content (TPC) and antioxidant capacity (AC) of Polygonum multiflorum Thunb. root using microwave-assisted extraction (MAE). The raw material used was Polygonum multiflorum Thunb. root powder. Five factors such as solvent type, solvent concentrations, solvent/material ratio, extraction time and microwave power were studied; TPC and AC values were determined by the Folin-Ciocalteu method and DPPH free radical scavenging activity measurement, respectively. In addition, studies involved assaying the HPLC test of extracts and SEM of samples. Optimal results pointed to acetone as the solvent, acetone concentration of 60%, solvent/material ratio of 40/1 (v/w), extraction time of 5 mins and microwave power of 127 W. TPC and AC obtained were approximates 44.3 ±0.13 mg GAE/g DW and 341.26 ±1.54 μmol TE/g DW, respectively. The effect of microwaving on the cell destruction of Polygonum multiflorum Thunb. root was observed by scanning electron microscopy (SEM). Some phenolic compounds were determined by the HPLC method, for instance, gallic acid, catechin and resveratrol. These factors significantly affected TPC and AC. We can use acetone as a solvent with microwave-assisted extraction to achieve the best result.

Protein 4.1 and its interaction with other cytoskeletal proteins in Xenopus laevis oogenesis.

PubMed

Carotenuto, Rosa; Petrucci, Tamara C; Correas, Isabel; Vaccaro, Maria C; De Marco, Nadia; Dale, Brian; Wilding, Martin

2009-06-01

In human red blood cells, protein 4.1 (4.1R) is an 80-kDa polypeptide that stabilizes the spectrin-actin network and anchors it to the plasma membrane. In non-erythroid cells there is a great variety of 4.1R isoforms, mainly generated by alternative pre-mRNA splicing, which localize at various intracellular sites, including the nucleus. We studied protein 4.1R distribution in relation to beta-spectrin, actin and cytokeratin during Xenopus oogenesis. Immunoprecipitation experiments indicate that at least two isoforms of protein 4.1R are present in Xenopus laevis oocytes: a 56-kDa form in the cytoplasm and a 37-kDa form in the germinal vesicle (GV). Antibodies to beta-spectrin reveal two bands of 239 and 100 kDa in the cytoplasm. Coimmunoprecipitation experiments indicate that both the 37- and 56-kDa isoforms of protein 4.1R associate with the 100-kDa isoform of beta-spectrin. Moreover, the 56-kDa form coimmunoprecipitates with a cytokeratin of the same molecular weight. Confocal immunolocalization shows that protein 4.1R distribution is in the peripheral cytoplasm, in the mitochondrial cloud (MC) and in the GV of previtellogenic oocytes. In the cytoplasm of vitellogenic oocytes, a loose network of fibers stained by the anti-protein 4.1R antibody spreads across the cytoplasm. beta-Spectrin has a similar distribution. Protein 4.1R was found to colocalize with actin in the cortex of oocytes in the form of fluorescent dots. Double immunolocalization of protein 4.1R and cytokeratin depicts two separate networks that overlap throughout the whole cytoplasm. Protein 4.1R filaments partially colocalize with cytokeratin in both the animal and vegetal hemispheres. We hypothesize that protein 4.1R could function as a linker protein between cytokeratin and the actin-based cytoskeleton.

An Integrated Specification and Verification Environment for Component-Based Architectures of Large-Scale Distributed Systems

DTIC Science & Technology

2009-05-26

Interrupt HW Interrupt DFS Dynamic Frequency Selection TPC Transmit Power Control r- MPX Hub ! i j I Power Supply Init/Reset A/D...values of several variables: from IN_0_DAT when the Mailbox forwards data supplied by Client 0, from OUT1DAT when the conditions on the ready flags are...logically implies ip y 0, and also <j> logically implies ip y <p; (b) if for all i we have that cj>t+1 is of the form (p% y ip, then the chain

Properties and function of nephrocalcin: mechanism of kidney stone inhibition or promotion.

PubMed

Nakagawa, Y

1997-03-01

Nephrocalcin (NC), an acidic glycoprotein with molecular weight 14,000, is present in urine and prevents kidney stone formation. Histoimmunochemical staining shows that NC is localized in the proximal tubles in kidneys. Isolated NC from mammalian urine, revealed at least 4 isoforms of NC (we call these isoforms NC-A, NC-B, NC-C, and NC-D in the order of elution) during DEAE cellulose column chromatography with a linear gradient of NaCl elution step. Non-stone forming people excrete more NC-A and NC-B isoforms in urine; however, more NC-C and NC-D isoforms were found in stone formers' urine. When the organic matrix was extracted from surgically removed calcium oxalate kidney stones, we found greater quantities of NC-C and NC-D isoforms than those of NC-A and NC-B isoforms. Amino acid compositions and carbohydrate contents of these 4 isoforms were similar with the exception of the gamma-carboxyglutamic acid (GLA) residues. Only the NC-A and NC-B isoforms contained residues of GLA. There were more phosphate residues present in NC-C and NC-D than in NC-A and NC-B. Upon removal of phosphate residues by alkaline phosphatase, NC-C eluted at the same salt concentrations as NC-A eluted. This indicates that the backbone protein could be similar, but the NC-C isoform is modified by excess phosphate residues. Surface tension measurements using a Lauda film balance indicated that NC-A and -B were strongly amphiphilic while NC-C and -D were less amphiphilic. NC-A has an elongated shape, and occupies a smaller area per molecule; whereas NC-C is a bulky molecule. Using NC-A as a model of a "good" inhibitor and NC-C as a model of a "poor" inhibitor, both bound with 4 atoms of Ca2+ per molecule as investigated by equilibrium dialysis method, 31P-NMR, and electron spin resonance spectrometry. Isoforms A and B changed their conformation upon Ca2+ binding, but C and D did not change their conformation. All these observations suggest that isoforms A and B are strong inhibitors of calcium oxalate monohydrate (COM) crystal growth and aggregation. However, isoforms C and D act as promotors for COM crystal growth-kidney stone formation. Measuring the amount of NC in urine from renal cell carcinoma patients and from NC isolated from a supernatant of a primary renal cell carcinoma cells demonstrated the amount of NC increased with disease progression.

Simulation of double beta decay in the ''SeXe'' TPC

NASA Astrophysics Data System (ADS)

Mauger, F.

2007-04-01

In 2004, the NEMO collaboration has started some preliminary studies for a next-generation double beta decay experiment: SuperNEMO. The possibility to use a large gaseous TPC has been investigated using simulation and extrapolation of former experiments. In this talk, I report on the reasons why such techniques have not been selected in 2004 and led the NEMO collaboration to reuse the techniques implemented within the NEMO3 detector.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Averyanov, A. V.; Bajajin, A. G.; Chepurnov, V. F.

The time-projection chamber (TPC) is the main tracking detector in the MPD/NICA. The information on charge-particle tracks in the TPC is registered by the MWPG with cathode pad readout. The frontend electronics (FEE) are developed with use of modern technologies such as application specific integrated circuits (ASIC), field-programmable gate arrays (FPGA), and data transfer to a concentrator via a fast optical interface. The main parameters of the FEE are as follows: total number of channels, ∼95 000; data stream from the whole TPC, 5 GB/s; low power consumption, less than 100 mW/ch; signal to noise ratio (S/N), 30; equivalent noisemore » charge (ENC), <1000e{sup –} (C{sub in} = 10–20 pF); and zero suppression (pad signal rejection ∼90%). The article presents the status of the readout chamber construction and the data acquisition system. The results of testing FEE prototypes are presented.« less

A melanosomal two-pore sodium channel regulates pigmentation

PubMed Central

Bellono, Nicholas W.; Escobar, Iliana E.; Oancea, Elena

2016-01-01

Intracellular organelles mediate complex cellular functions that often require ion transport across their membranes. Melanosomes are organelles responsible for the synthesis of the major mammalian pigment melanin. Defects in melanin synthesis result in pigmentation defects, visual deficits, and increased susceptibility to skin and eye cancers. Although genes encoding putative melanosomal ion transporters have been identified as key regulators of melanin synthesis, melanosome ion transport and its contribution to pigmentation remain poorly understood. Here we identify two-pore channel 2 (TPC2) as the first reported melanosomal cation conductance by directly patch-clamping skin and eye melanosomes. TPC2 has been implicated in human pigmentation and melanoma, but the molecular mechanism mediating this function was entirely unknown. We demonstrate that the vesicular signaling lipid phosphatidylinositol bisphosphate PI(3,5)P2 modulates TPC2 activity to control melanosomal membrane potential, pH, and regulate pigmentation. PMID:27231233

Design and implementation of wire tension measurement system for MWPCs used in the STAR iTPC upgrade

DOE PAGES

Wang, Xu; Shen, Fuwang; Wang, Shuai; ...

2017-04-06

The STAR experiment at RHIC is planning to upgrade the Time Projection Chamber which lies at the heart of the detector. We have designed an instrument to measure the tension of the wires in the multi-wire proportional chambers (MWPCs) which will be used in the TPC upgrade. The wire tension measurement system causes the wires to vibrate and then it measures the fundamental frequency of the oscillation via a laser based optical platform. The platform can scan the entire wire plane, automatically, in a single run and obtain the wire tension on each wire with high precision. In this paper,more » the details about the measurement method and the system setup will be described. In addition, the test results for a prototype MWPC to be used in the STAR-iTPC upgrade will be presented.« less

Time Projection Chamber Polarimeters for X-ray Astrophysics

NASA Astrophysics Data System (ADS)

Hill, Joanne; Black, Kevin; Jahoda, Keith

2015-04-01

Time Projection Chamber (TPC) based X-ray polarimeters achieve the sensitivity required for practical and scientifically significant astronomical observations, both galactic and extragalactic, with a combination of high analyzing power and good quantum efficiency. TPC polarimeters at the focus of an X-ray telescope have low background and large collecting areas providing the ability to measure the polarization properties of faint persistent sources. TPCs based on drifting negative ions rather than electrons permit large detector collecting areas with minimal readout electronics enabling wide field of view polarimeters for observing unpredictable, bright transient sources such as gamma-ray bursts. We described here the design and expected performance of two different TPC polarimeters proposed for small explorer missions: The PRAXyS (Polarimetry of Relativistic X-ray Sources) X-ray Polarimeter Instrument, optimized for observations of faint persistent sources and the POET (Polarimetry of Energetic Transients) Low Energy Polarimeter, designed to detect and measure bright transients. also NASA/GSFC.

Comparison of different strategies for soybean antioxidant extraction.

PubMed

Chung, Hyun; Ji, Xiangming; Canning, Corene; Sun, Shi; Zhou, Kequan

2010-04-14

Three extraction strategies including Soxhlet extraction, conventional solid-liquid extraction, and ultrasonic-assisted extraction (UAE) were compared for their efficiency to extract phenolic antioxidants from Virginia-grown soybean seeds. Five extraction solvents were evaluated in UAE and the conventional extraction. The soybean extracts were compared for their total phenolic contents (TPC), oxygen radical absorbance capacity (ORAC), and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH(*)) scavenging activities. The results showed that UAE improved the extraction of soybean phenolic compounds by >54% compared to the conventional and Soxhlet extractions. Among the tested solvents, 50% acetone was the most efficient for extracting soybean phenolic compounds. There was no significant correlation between the TPC and antioxidant activities of the soybean extracts. The extracts prepared by 70% ethanol had the highest ORAC values. Overall, UAE with 50% acetone or 70% ethanol is recommended for extracting soybean antioxidants on the basis of the TPC and ORAC results.

Design and implementation of wire tension measurement system for MWPCs used in the STAR iTPC upgrade

NASA Astrophysics Data System (ADS)

Wang, Xu; Shen, Fuwang; Wang, Shuai; Feng, Cunfeng; Li, Changyu; Lu, Peng; Thomas, Jim; Xu, Qinghua; Zhu, Chengguang

2017-07-01

The STAR experiment at RHIC is planning to upgrade the Time Projection Chamber which lies at the heart of the detector. We have designed an instrument to measure the tension of the wires in the multi-wire proportional chambers (MWPCs) which will be used in the TPC upgrade. The wire tension measurement system causes the wires to vibrate and then it measures the fundamental frequency of the oscillation via a laser based optical platform. The platform can scan the entire wire plane, automatically, in a single run and obtain the wire tension on each wire with high precision. In this paper, the details about the measurement method and the system setup will be described. In addition, the test results for a prototype MWPC to be used in the STAR-iTPC upgrade will be presented.

Benchmarking Using Basic DBMS Operations

NASA Astrophysics Data System (ADS)

Crolotte, Alain; Ghazal, Ahmad

The TPC-H benchmark proved to be successful in the decision support area. Many commercial database vendors and their related hardware vendors used these benchmarks to show the superiority and competitive edge of their products. However, over time, the TPC-H became less representative of industry trends as vendors keep tuning their database to this benchmark-specific workload. In this paper, we present XMarq, a simple benchmark framework that can be used to compare various software/hardware combinations. Our benchmark model is currently composed of 25 queries that measure the performance of basic operations such as scans, aggregations, joins and index access. This benchmark model is based on the TPC-H data model due to its maturity and well-understood data generation capability. We also propose metrics to evaluate single-system performance and compare two systems. Finally we illustrate the effectiveness of this model by showing experimental results comparing two systems under different conditions.

HPLC-UV-ESI-MS analysis of phenolic compounds and antioxidant properties of Hypericum undulatum shoot cultures and wild-growing plants.

PubMed

Rainha, Nuno; Koci, Kamila; Coelho, Ana Varela; Lima, Elisabete; Baptista, José; Fernandes-Ferreira, Manuel

2013-02-01

LC-UV and LC-MS analysis were used to study the phenolic composition of water extracts of Hypericum undulatum (HU) shoot cultures and wild-growing (WG) plants. Total phenolic content (TPC), determined using the Folin-Ciocalteu assay, and the antioxidant activity measured by two complementary methods were also performed for each sample. Mass spectrometry revealed several phenolics acids with quinic acid moieties, flavonols, mostly quercetin, luteolin and apigenin glycosides, flavan-3-ols (catechin and epicatechin) and the xanthonoid mangiferin. Differences in phenolic composition profile and TPC were found between the samples. The major phenolic in HU culture-growing (CG) samples is chlorogenic acid, followed by epicatechin, quercitrin and isoquercitrin. The WG plants presents hyperoside as the main phenolic, followed by isoquercitrin, chlorogenic acid and quercetin. The TPC and antioxidant activity were higher in samples from WG plants. Copyright © 2012 Elsevier Ltd. All rights reserved.

The darkside multiton detector for the direct dark matter search

DOE PAGES

Aalseth, C. E.; Agnes, P.; Alton, A.; ...

2015-01-01

Although the existence of dark matter is supported by many evidences, based on astrophysical measurements, its nature is still completely unknown. One major candidate is represented by weakly interacting massive particles (WIMPs), which could in principle be detected through their collisions with ordinary nuclei in a sensitive target, producing observable low-energy (<100 keV) nuclear recoils. The DarkSide program aims at the WIPMs detection using a liquid argon time projection chamber (LAr-TPC). In this paper we quickly review the DarkSide program focusing in particular on the next generation experiment DarkSide-G2, a 3.6-ton LAr-TPC. The different detector components are described as wellmore » as the improvements needed to scale the detector from DarkSide-50 (50 kg LAr-TPC) up to DarkSide-G2. Finally, the preliminary results on background suppression and expected sensitivity are presented.« less

Effect of Microwave Vacuum Drying on the Drying Characteristics, Color, Microstructure, and Antioxidant Activity of Green Coffee Beans.

PubMed

Dong, Wenjiang; Cheng, Ke; Hu, Rongsuo; Chu, Zhong; Zhao, Jianping; Long, Yuzhou

2018-05-11

The aim of this study is to investigate the effect of microwave vacuum drying (MVD) on the drying characteristics and quality attributes of green coffee beans. We specifically focused on the effective moisture diffusion coefficient ( D eff ), surface temperature, glass transition temperature ( T g ), water state, and microstructure. The kinetics of color changes during drying, total phenolic content (TPC), and antioxidant activity (DPPH, FRAP, and ABTS) were also characterized. Microwave power during MVD affected the porosity of coffee beans, their color, TPC, and antioxidant activity. The Allometric 1 model was the most suitable for simulating surface temperature rise kinetics. Thermal processing of green coffee beans resulted in increased b* , L* , Δ E , and TPC values, and greater antioxidant capacity. These findings may provide a theoretical reference for the technical improvement, mechanisms of flavor compound formation, and quality control of dried green coffee beans.

MAISTAS: a tool for automatic structural evaluation of alternative splicing products.

PubMed

Floris, Matteo; Raimondo, Domenico; Leoni, Guido; Orsini, Massimiliano; Marcatili, Paolo; Tramontano, Anna

2011-06-15

Analysis of the human genome revealed that the amount of transcribed sequence is an order of magnitude greater than the number of predicted and well-characterized genes. A sizeable fraction of these transcripts is related to alternatively spliced forms of known protein coding genes. Inspection of the alternatively spliced transcripts identified in the pilot phase of the ENCODE project has clearly shown that often their structure might substantially differ from that of other isoforms of the same gene, and therefore that they might perform unrelated functions, or that they might even not correspond to a functional protein. Identifying these cases is obviously relevant for the functional assignment of gene products and for the interpretation of the effect of variations in the corresponding proteins. Here we describe a publicly available tool that, given a gene or a protein, retrieves and analyses all its annotated isoforms, provides users with three-dimensional models of the isoform(s) of his/her interest whenever possible and automatically assesses whether homology derived structural models correspond to plausible structures. This information is clearly relevant. When the homology model of some isoforms of a gene does not seem structurally plausible, the implications are that either they assume a structure unrelated to that of the other isoforms of the same gene with presumably significant functional differences, or do not correspond to functional products. We provide indications that the second hypothesis is likely to be true for a substantial fraction of the cases. http://maistas.bioinformatica.crs4.it/.

Myelin management by the 18.5–kDa and 21.5–kDa classic myelin basic protein isoforms

PubMed Central

Harauz, George; Boggs, Joan M.

2013-01-01

The classic myelin basic protein (MBP) splice isoforms range in nominal molecular mass from 14 to 21.5 kDa, and arise from the gene in the oligodendrocyte lineage (Golli) in maturing oligodendrocytes. The 18.5-kDa isoform that predominates in adult myelin adheres the cytosolic surfaces of oligodendrocyte membranes together, and forms a two-dimensional molecular sieve restricting protein diffusion into compact myelin. However, this protein has additional roles including cytoskeletal assembly and membrane extension, binding to SH3-domains, participation in Fyn-mediated signaling pathways, sequestration of phosphoinositides, and maintenance of calcium homeostasis. Of the diverse post-translational modifications of this isoform, phosphorylation is the most dynamic, and modulates 18.5-kDa MBP’s protein-membrane and protein-protein interactions, indicative of a rich repertoire of functions. In developing and mature myelin, phosphorylation can result in microdomain or even nuclear targeting of the protein, supporting the conclusion that 18.5-kDa MBP has significant roles beyond membrane adhesion. The full-length, early-developmental 21.5-kDa splice isoform is predominantly karyophilic due to a non-traditional P-Y nuclear localization signal, with effects such as promotion of oligodendrocyte proliferation. We discuss in vitro and recent in vivo evidence for multifunctionality of these classic basic proteins of myelin, and argue for a systematic evaluation of the temporal and spatial distributions of these protein isoforms, and their modified variants, during oligodendrocyte differentiation. PMID:23398367

A Short Isoform of Human Cytomegalovirus US3 Functions as a Dominant Negative Inhibitor of the Full-Length Form

PubMed Central

Shin, Jinwook; Park, Boyoun; Lee, Sungwook; Kim, Youngkyun; Biegalke, Bonita J.; Kang, Seongman; Ahn, Kwangseog

2006-01-01

Human cytomegalovirus encodes four unique short (US) region proteins, each of which is independently sufficient for causing the down-regulation of major histocompatibility complex (MHC) class I molecules on the cell surface. This down-regulation enables infected cells to evade recognition by cytotoxic T lymphocytes (CTLs) but makes them vulnerable to lysis by natural killer (NK) cells, which lyse those cells that lack MHC class I molecules. The 22-kDa US3 glycoprotein is able to down-regulate the surface expression of MHC class I molecules by dual mechanisms: direct endoplasmic reticulum retention by physical association and/or tapasin inhibition. The alternative splicing of the US3 gene generates two additional products, including 17-kDa and 3.5-kDa truncated isoforms; however, the functional significance of these isoforms during viral infection is unknown. Here, we describe a novel mode of self-regulation of US3 function that uses the endogenously produced truncated isoform. The truncated isoform itself neither binds to MHC class I molecules nor prevents the full-length US3 from interacting with MHC class I molecules. Instead, the truncated isoform associates with tapasin and competes with full-length US3 for binding to tapasin; thus, it suppresses the action of US3 that causes the disruption of the function of tapasin. Our results indicate that the truncated isoform of the US3 locus acts as a dominant negative regulator of full-length US3 activity. These data reflect the manner in which the virus has developed temporal survival strategies during viral infection against immune surveillance involving both CTLs and NK cells. PMID:16699020

Molecular Analysis of Collagen XVIII Reveals Novel Mutations, Presence of a Third Isoform, and Possible Genetic Heterogeneity in Knobloch Syndrome

PubMed Central

Suzuki, O. T.; Sertié, A. L.; Der Kaloustian, V. M.; Kok, F.; Carpenter, M.; Murray, J.; Czeizel, A. E.; Kliemann, S. E.; Rosemberg, S.; Monteiro, M.; Olsen, B. R.; Passos-Bueno, M. R.

2002-01-01

Knobloch syndrome (KS) is a rare disease characterized by severe ocular alterations, including vitreoretinal degeneration associated with retinal detachment and occipital scalp defect. The responsible gene, COL18A1, has been mapped to 21q22.3, and, on the basis of the analysis of one family, we have demonstrated that a mutation affecting only one of the three COL18A1 isoforms causes this phenotype. We report here the results of the screening of both the entire coding region and the exon-intron boundaries of the COL18A1 gene (which includes 43 exons), in eight unrelated patients with KS. Besides 20 polymorphic changes, we identified 6 different pathogenic changes in both alleles of five unrelated patients with KS (three compound heterozygotes and two homozygotes). All are truncating mutations leading to deficiency of one or all collagen XVIII isoforms and endostatin. We have verified that, in exon 41, the deletion c3514-3515delCT, found in three unrelated alleles, is embedded in different haplotypes, suggesting that this mutation has occurred more than once. In addition, our results provide evidence of nonallelic genetic heterogeneity in KS. We also show that the longest human isoform (NC11-728) is expressed in several tissues (including the human eye) and that lack of either the short variant or all of the collagen XVIII isoforms causes similar phenotypes but that those patients who lack all forms present more-severe ocular alterations. Despite the small sample size, we found low endostatin plasma levels in those patients with mutations leading to deficiency of all isoforms; in addition, it seems that absence of all collagen XVIII isoforms causes predisposition to epilepsy. PMID:12415512

The novel protein kinase C epsilon isoform at the adult neuromuscular synapse: location, regulation by synaptic activity-dependent muscle contraction through TrkB signaling and coupling to ACh release.

PubMed

Obis, Teresa; Besalduch, Núria; Hurtado, Erica; Nadal, Laura; Santafe, Manel M; Garcia, Neus; Tomàs, Marta; Priego, Mercedes; Lanuza, Maria A; Tomàs, Josep

2015-02-10

Protein kinase C (PKC) regulates a variety of neural functions, including neurotransmitter release. Although various PKC isoforms can be expressed at the synaptic sites and specific cell distribution may contribute to their functional diversity, little is known about the isoform-specific functions of PKCs in neuromuscular synapse. The present study is designed to examine the location of the novel isoform nPKCε at the neuromuscular junction (NMJ), their synaptic activity-related expression changes, its regulation by muscle contraction, and their possible involvement in acetylcholine release. We use immunohistochemistry and confocal microscopy to demonstrate that the novel isoform nPKCε is exclusively located in the motor nerve terminals of the adult rat NMJ. We also report that electrical stimulation of synaptic inputs to the skeletal muscle significantly increased the amount of nPKCε isoform as well as its phosphorylated form in the synaptic membrane, and muscle contraction is necessary for these nPKCε expression changes. The results also demonstrate that synaptic activity-induced muscle contraction promotes changes in presynaptic nPKCε through the brain-derived neurotrophic factor (BDNF)-mediated tyrosine kinase receptor B (TrkB) signaling. Moreover, nPKCε activity results in phosphorylation of the substrate MARCKS involved in actin cytoskeleton remodeling and related with neurotransmission. Finally, blocking nPKCε with a nPKCε-specific translocation inhibitor peptide (εV1-2) strongly reduces phorbol ester-induced ACh release potentiation, which further indicates that nPKCε is involved in neurotransmission. Together, these results provide a mechanistic insight into how synaptic activity-induced muscle contraction could regulate the presynaptic action of the nPKCε isoform and suggest that muscle contraction is an important regulatory step in TrkB signaling at the NMJ.

A short isoform of human cytomegalovirus US3 functions as a dominant negative inhibitor of the full-length form.

PubMed

Shin, Jinwook; Park, Boyoun; Lee, Sungwook; Kim, Youngkyun; Biegalke, Bonita J; Kang, Seongman; Ahn, Kwangseog

2006-06-01

Human cytomegalovirus encodes four unique short (US) region proteins, each of which is independently sufficient for causing the down-regulation of major histocompatibility complex (MHC) class I molecules on the cell surface. This down-regulation enables infected cells to evade recognition by cytotoxic T lymphocytes (CTLs) but makes them vulnerable to lysis by natural killer (NK) cells, which lyse those cells that lack MHC class I molecules. The 22-kDa US3 glycoprotein is able to down-regulate the surface expression of MHC class I molecules by dual mechanisms: direct endoplasmic reticulum retention by physical association and/or tapasin inhibition. The alternative splicing of the US3 gene generates two additional products, including 17-kDa and 3.5-kDa truncated isoforms; however, the functional significance of these isoforms during viral infection is unknown. Here, we describe a novel mode of self-regulation of US3 function that uses the endogenously produced truncated isoform. The truncated isoform itself neither binds to MHC class I molecules nor prevents the full-length US3 from interacting with MHC class I molecules. Instead, the truncated isoform associates with tapasin and competes with full-length US3 for binding to tapasin; thus, it suppresses the action of US3 that causes the disruption of the function of tapasin. Our results indicate that the truncated isoform of the US3 locus acts as a dominant negative regulator of full-length US3 activity. These data reflect the manner in which the virus has developed temporal survival strategies during viral infection against immune surveillance involving both CTLs and NK cells.

Differential tissue distribution of tryptophan hydroxylase isoforms 1 and 2 as revealed with monospecific antibodies.

PubMed

Sakowski, Stacey A; Geddes, Timothy J; Thomas, David M; Levi, Edi; Hatfield, James S; Kuhn, Donald M

2006-04-26

Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in the synthesis of the neurotransmitter serotonin. Once thought to be a single-gene product, TPH is now known to exist in two isoforms-TPH1 is found in the pineal and gut, and TPH2 is selectively expressed in brain. Heretofore, probes used for localization of TPH protein or mRNA could not distinguish between the TPH isoforms because of extensive homology shared by them at the nucleotide and amino acid level. We have produced monospecific polyclonal antibodies against TPH1 and TPH2 using peptide antigens from nonoverlapping sequences in the respective proteins. These antibodies allow the differentiation of TPH1 and TPH2 upon immunoblotting, immunoprecipitation, and immunocytochemical staining of tissue sections from brain and gut. TPH1 and TPH2 antibodies do not cross-react with either tyrosine hydroxylase or phenylalanine hydroxylase. Analysis of mouse tissues confirms that TPH1 is the predominant form expressed in pineal gland and in P815 mastocytoma cells with a molecular weight of 51 kDa. TPH2 is the predominant enzyme form expressed in brain extracts from mesencephalic tegmentum, striatum, and hippocampus with a molecular weight of 56 kDa. Antibody specificity against TPH1 and TPH2 is retained across mouse, rat, rabbit, primate, and human tissues. Antibodies that distinguish between the isoforms of TPH will allow studies of the differential regulation of their expression in brain and periphery.

DNA polymerase ι: The long and the short of it!

PubMed

Frank, Ekaterina G; McLenigan, Mary P; McDonald, John P; Huston, Donald; Mead, Samantha; Woodgate, Roger

2017-10-01

The cDNA encoding human DNA polymerase ι (POLI) was cloned in 1999. At that time, it was believed that the POLI gene encoded a protein of 715 amino acids. Advances in DNA sequencing technologies led to the realization that there is an upstream, in-frame initiation codon that would encode a DNA polymerase ι (polι) protein of 740 amino acids. The extra 25 amino acid region is rich in acidic residues (11/25) and is reasonably conserved in eukaryotes ranging from fish to humans. As a consequence, the curated Reference Sequence (RefSeq) database identified polι as a 740 amino acid protein. However, the existence of the 740 amino acid polι has never been shown experimentally. Using highly specific antibodies to the 25 N-terminal amino acids of polι, we were unable to detect the longer 740 amino acid (ι-long) isoform in western blots. However, trace amounts of the ι-long isoform were detected after enrichment by immunoprecipitation. One might argue that the longer isoform may have a distinct biological function, if it exhibits significant differences in its enzymatic properties from the shorter, well-characterized 715 amino acid polι. We therefore purified and characterized recombinant full-length (740 amino acid) polι-long and compared it to full-length (715 amino acid) polι-short in vitro. The metal ion requirements for optimal catalytic activity differ slightly between ι-long and ι-short, but under optimal conditions, both isoforms exhibit indistinguishable enzymatic properties in vitro. We also report that like ι-short, the ι-long isoform can be monoubiquitinated and polyubiuquitinated in vivo, as well as form damage induced foci in vivo. We conclude that the predominant isoform of DNA polι in human cells is the shorter 715 amino acid protein and that if, or when, expressed, the longer 740 amino acid isoform has identical properties to the considerably more abundant shorter isoform. Published by Elsevier B.V.

Metal-core pad-plane development for ACTAR TPC

NASA Astrophysics Data System (ADS)

Giovinazzo, J.; Pibernat, J.; Goigoux, T.; de Oliveira, R.; Grinyer, G. F.; Huss, C.; Mauss, B.; Pancin, J.; Pedroza, J. L.; Rebii, A.; Roger, T.; Rosier, P.; Saillant, F.; Wittwer, G.

2018-06-01

With the recent development of active targets and time projection chambers (ACTAR TPC) as detectors for fundamental nuclear physics experiments, the need arose for charge collection planes with a high density of readout channels. In order to fulfill the mechanical constraints for the ACTAR TPC device, we designed a pad-plane based on a metal-core circuit with an conceptually simple design and routing for signal readout, named FAKIR (in reference to a fakir bed of nails). A test circuit has been equipped with a micro mesh gaseous structure (micromegas) for signal amplification and a dedicated readout electronics. Test measurements have been performed with an 55Fe X-ray source giving an intrinsic energy resolution (FWHM) of 22 ± 1% at 5 . 9 keV, and with a 3-alpha source for which a resolution of about 130 ± 20 keV at 4 . 8 MeV has been estimated. The pad-plane has been mounted into a reduced size demonstrator version of the ACTAR TPC detector, in order to illustrate charged particle track reconstruction. The tests preformed with the X-ray and the 3-alpha sources shows that results obtained from pads signals are comparable to the intrinsic result from the micro-mesh signal. In addition, a simple alpha particle tracks analysis is performed to demonstrate that the pad plane allows a precise reconstruction of the direction and length of the trajectories.

Optical readout of a two phase liquid argon TPC using CCD camera and THGEMs

NASA Astrophysics Data System (ADS)

Mavrokoridis, K.; Ball, F.; Carroll, J.; Lazos, M.; McCormick, K. J.; Smith, N. A.; Touramanis, C.; Walker, J.

2014-02-01

This paper presents a preliminary study into the use of CCDs to image secondary scintillation light generated by THick Gas Electron Multipliers (THGEMs) in a two phase LAr TPC. A Sony ICX285AL CCD chip was mounted above a double THGEM in the gas phase of a 40 litre two-phase LAr TPC with the majority of the camera electronics positioned externally via a feedthrough. An Am-241 source was mounted on a rotatable motion feedthrough allowing the positioning of the alpha source either inside or outside of the field cage. Developed for and incorporated into the TPC design was a novel high voltage feedthrough featuring LAr insulation. Furthermore, a range of webcams were tested for operation in cryogenics as an internal detector monitoring tool. Of the range of webcams tested the Microsoft HD-3000 (model no:1456) webcam was found to be superior in terms of noise and lowest operating temperature. In ambient temperature and atmospheric pressure 1 ppm pure argon gas, the THGEM gain was ≈ 1000 and using a 1 msec exposure the CCD captured single alpha tracks. Successful operation of the CCD camera in two-phase cryogenic mode was also achieved. Using a 10 sec exposure a photograph of secondary scintillation light induced by the Am-241 source in LAr has been captured for the first time.

Intra-laboratory validation of microplate methods for total phenolic content and antioxidant activity on polyphenolic extracts, and comparison with conventional spectrophotometric methods.

PubMed

Bobo-García, Gloria; Davidov-Pardo, Gabriel; Arroqui, Cristina; Vírseda, Paloma; Marín-Arroyo, María R; Navarro, Montserrat

2015-01-01

Total phenolic content (TPC) and antioxidant activity (AA) assays in microplates save resources and time, therefore they can be useful to overcome the fact that the conventional methods are time-consuming, labour intensive and use large amounts of reagents. An intra-laboratory validation of the Folin-Ciocalteu microplate method to measure TPC and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) microplate method to measure AA was performed and compared with conventional spectrophotometric methods. To compare the TPC methods, the confidence intervals of a linear regression were used. In the range of 10-70 mg L(-1) of gallic acid equivalents (GAE), both methods were equivalent. To compare the AA methodologies, the F-test and t-test were used in a range from 220 to 320 µmol L(-1) of Trolox equivalents. Both methods had homogeneous variances, and the means were not significantively different. The limits of detection and quantification for the TPC microplate method were 0.74 and 2.24 mg L(-1) GAE and for the DPPH 12.07 and 36.58 µmol L(-1) of Trolox equivalents. The relative standard deviation of the repeatability and reproducibility for both microplate methods were ≤ 6.1%. The accuracy ranged from 88% to 100%. The microplate and the conventional methods are equals in a 95% confidence level. © 2014 Society of Chemical Industry.

Triptycene-Based Microporous Cyanate Resins for Adsorption/Separations of Benzene/Cyclohexane and Carbon Dioxide Gas.

PubMed

Deng, Gaoyang; Wang, Zhonggang

2017-11-29

Triptycene-based cyanate monomers 2,6,14-tricyanatotriptycene (TPC) and 2,6,14-tris(4-cyanatophenyl)triptycene (TPPC) that contain different numbers of benzene rings per molecule were synthesized, from which two microporous cyanate resins PCN-TPC and PCN-TPPC were prepared. Of interest is the observation that the two polymers have very similar porosity parameters, but PCN-TPPC uptakes considerably higher benzene (77.8 wt %) than PCN-TPC (17.6 wt %) at room temperature since the higher concentration of phenyl groups in PCN-TPPC enhances the π-π interaction with benzene molecules. Besides, the adsorption capacity of benzene in PCN-TPPC is dramatically 7 times as high as that of cyclohexane. Contrary to the adsorption of organic vapors, at 273 K and 1.0 bar, PCN-TPC with more heteroatoms in the network skeleton displays larger uptake of CO 2 and higher CO 2 /N 2 selectivity (16.4 wt %, 60) than those of PCN-TPPC (14.0 wt %, 39). The excellent and unique adsorption properties exhibit potential applications in the purification of small molecular organic hydrocarbons, e.g., separation of benzene from benzene/cyclohexane mixture as well as CO 2 capture from flue gas. Moreover, the results are helpful for deeply understanding the effect of porous and chemical structures on the adsorption properties of organic hydrocarbons and CO 2 gas.

Integration of the ATLAS FE-I4 Pixel Chip in the Mini Time Projection Chamber

NASA Astrophysics Data System (ADS)

Lopez-Thibodeaux, Mayra; Garcia-Sciveres, Maurice; Kadyk, John; Oliver-Mallory, Kelsey

2013-04-01

This project deals with development of readout for a Time Projection Chamber (TPC) prototype. This is a type of detector proposed for direct detection of dark matter (WIMPS) with direction information. The TPC is a gaseous charged particle tracking detector composed of a field cage and a gas avalanche detector. The latter is made of two Gas Electron Multipliers in series, illuminating a pixel readout integrated circuit, which measures the distribution in position and time of the output charge. We are testing the TPC prototype, filled with ArCO2 gas, using a Fe-55 x-ray source and cosmic rays. The present prototype uses an FE-I3 chip for readout. This chip was developed about 10 years ago and is presently in use within the ATLAS pixel detector at the LHC. The aim of this work is to upgrade the TPC prototype to use an FE-I4 chip. The FE-I4 has an active area of 336 mm^2 and 26880 pixels, over nine times the number of pixels in the FE-I3 chip, and an active area about six times as much. The FE-I4 chip represents the state of the art of pixel detector readout, and is presently being used to build an upgrade of the ATLAS pixel detector.

Tropomodulins are negative regulators of neurite outgrowth

PubMed Central

Fath, Thomas; Fischer, Robert S.; Dehmelt, Leif; Halpain, Shelley; Fowler, Velia M.

2010-01-01

Regulation of the actin cytoskeleton is critical for neurite formation. Tropomodulins (Tmods) regulate polymerization at actin filament pointed ends. Previous experiments using a mouse model deficient for the neuron specific isoform Tmod2 suggested a role for Tmods in neuronal function by impacting processes underlying learning and memory. However, the role of Tmods in neuronal function on the cellular level remains unknown. Immunofluorescence localization of the neuronal isoforms Tmod1 and Tmod2 in cultured rat primary hippocampal neurons revealed that Tmod1 is enriched along the proximal part of F-actin bundles in lamellipodia of spreading cells and in growth cones of extending neurites, while Tmod2 appears largely cytoplasmic. Functional analysis of these Tmod isoforms in a mouse neuroblastoma N2a cell line showed that knockdown of Tmod2 resulted in a significant increase in number of neurite-forming cells and in neurite length. While N2a cells compensated for Tmod2 knockdown by increasing Tmod1 levels, over-expression of exogenous Tmod1 had no effect on neurite outgrowth. Moreover, knockdown of Tmod1 increased the number of neurites formed per cell, without effect on number of neurite-forming cells or neurite length. Taken together, these results indicate that Tmod1 and Tmod2 have mechanistically distinct inhibitory roles in neurite formation, likely mediated via different effects on F-actin dynamics and via differential localizations during early neuritogenesis. PMID:21146252

Basal activity of GIRK5 isoforms.

PubMed

Salvador, Carolina; Mora, Silvia I; Ordaz, Benito; Antaramian, Anaid; Vaca, Luis; Escobar, Laura I

2003-02-14

G protein-coupled inwardly rectifying K(+) channels (GIRK or Kir3) form functional heterotetramers gated by Gbetagamma subunits. GIRK channels are critical for functions as diverse as heart rate modulation and neuronal post-synaptic inhibition. GIRK5 (Kir3.5) is the oocyte homologue of the mammalian GIRK subunits that conform the K(ACh) channel. It has been claimed that even when the oocytes express GIRK5 proteins they do not form functional channels. However, the GIRK5 gene shows three initiation sites that suggest the existence of three isoforms. In a previous work we demonstrated the functionality of homomultimers of the shortest isoform overexpressed in the own oocytes. Remarkably, the basal GIRK5-Delta25 inward currents were not coupled to the activation of a G-protein receptor in the oocytes. These results encouraged us to study this channel in another expression system. In this work we show that Sf21 insect cells can be successfully transfected with this channel. GIRK5-Delta25 homomultimers produce time-dependent inward currents only with GTPgammaS in the recording pipette. Therefore, alternative modes of stimulus input to heterotrimeric G-proteins should be present in the oocytes to account for these results.

Isolated pores dissected from human two-pore channel 2 are functional

PubMed Central

Penny, Christopher J.; Rahman, Taufiq; Sula, Altin; Miles, Andrew J.; Wallace, B. A.; Patel, Sandip

2016-01-01

Multi-domain voltage-gated ion channels appear to have evolved through sequential rounds of intragenic duplication from a primordial one-domain precursor. Whereas modularity within one-domain symmetrical channels is established, little is known about the roles of individual regions within more complex asymmetrical channels where the domains have undergone substantial divergence. Here we isolated and characterised both of the divergent pore regions from human TPC2, a two-domain channel that holds a key intermediate position in the evolution of voltage-gated ion channels. In HeLa cells, each pore localised to the ER and caused Ca2+ depletion, whereas an ER-targeted pore mutated at a residue that inactivates full-length TPC2 did not. Additionally, one of the pores expressed at high levels in E. coli. When purified, it formed a stable, folded tetramer. Liposomes reconstituted with the pore supported Ca2+ and Na+ uptake that was inhibited by known blockers of full-length channels. Computational modelling of the pore corroborated cationic permeability and drug interaction. Therefore, despite divergence, both pores are constitutively active in the absence of their partners and retain several properties of the wild-type pore. Such symmetrical ‘pore-only’ proteins derived from divergent channel domains may therefore provide tractable tools for probing the functional architecture of complex ion channels. PMID:27941820

Time- and isomer-resolved measurements of sequential addition of acetylene to the propargyl radical

DOE PAGES

Savee, John D.; Selby, Talitha M.; Welz, Oliver; ...

2015-10-06

Soot formation in combustion is a complex process in which polycyclic aromatic hydrocarbons (PAHs) are believed to play a critical role. Recent works concluded that three consecutive additions of acetylene (C 2H 2) to propargyl (C 3H 3) create a facile route to the PAH indene (C 9H 8). However, the isomeric forms of C 5H 5 and C 7H 7 intermediates in this reaction sequence are not known. We directly investigate these intermediates using time- and isomer-resolved experiments. Both the resonance stabilized vinylpropargyl ( vp-C 5H 5) and 2,4-cyclopentadienyl ( c-C 5H 5) radical isomers of C 5H 5more » are produced, with substantially different intensities at 800 K vs 1000 K. In agreement with literature master equation calculations, we find that c-C 5H 5 + C 2H 2 produces only the tropyl isomer of C 7H 7 ( tp-C 7H 7) below 1000 K, and that tp-C 7H 7 + C 2H 2 terminates the reaction sequence yielding C 9H 8 (indene) + H. Lastly, this work demonstrates a pathway for PAH formation that does not proceed through benzene.« less

Isolated pores dissected from human two-pore channel 2 are functional.

PubMed

Penny, Christopher J; Rahman, Taufiq; Sula, Altin; Miles, Andrew J; Wallace, B A; Patel, Sandip

2016-12-12

Multi-domain voltage-gated ion channels appear to have evolved through sequential rounds of intragenic duplication from a primordial one-domain precursor. Whereas modularity within one-domain symmetrical channels is established, little is known about the roles of individual regions within more complex asymmetrical channels where the domains have undergone substantial divergence. Here we isolated and characterised both of the divergent pore regions from human TPC2, a two-domain channel that holds a key intermediate position in the evolution of voltage-gated ion channels. In HeLa cells, each pore localised to the ER and caused Ca 2+ depletion, whereas an ER-targeted pore mutated at a residue that inactivates full-length TPC2 did not. Additionally, one of the pores expressed at high levels in E. coli. When purified, it formed a stable, folded tetramer. Liposomes reconstituted with the pore supported Ca 2+ and Na + uptake that was inhibited by known blockers of full-length channels. Computational modelling of the pore corroborated cationic permeability and drug interaction. Therefore, despite divergence, both pores are constitutively active in the absence of their partners and retain several properties of the wild-type pore. Such symmetrical 'pore-only' proteins derived from divergent channel domains may therefore provide tractable tools for probing the functional architecture of complex ion channels.

Bioactive compounds and antioxidant activity of wolfberry infusion

PubMed Central

Sun, Yujing; Rukeya, Japaer; Tao, Wenyang; Sun, Peilong; Ye, Xingqian

2017-01-01

An infusion of the wolfberry (Lycium barbarum L.) is a traditional Asian herbal tea. This is the most commonly consumed form of dried wolfberry worldwide, yet little scientific information on wolfberry infusions is available. We investigated the effects of making infusions with hot water on the color, the content of bioactive compounds (polysaccharides, polyphenols, flavonoids and carotenoids) and the antioxidant ability of wolfberry infusions. The contents of bioactive compounds and the antioxidant activity of a wolfberry infusion increased with increased infusion temperature and time. Total polysaccharides content (TPOC), total polyphenols (TPC), total flavonoids (TFC) and total carotenoids contents (TCC) were important for determining the antioxidant capacity of wolfberry infusions with the contribution to antioxidant activity in the order TPC > TFC > TCC > TPOC. Hierarchical cluster analysis indicated preparation conditions of 100 °C for 1~3 h, 90 °C for 2~3 h and 80 °C for 2.5~3 h were equivalent as regards the value of TPC, TPOC, TFC, TCC, FRAP, DPPH and ABTS. The results of this study suggest the length of time of making a wolfberry infusion in actual real life practice is too short and different dietary habits associated with the intake of wolfberry infusion might provide the same bioactive nutrients. PMID:28102295

Antioxidant and antibacterial activity of Rhoeo spathacea (Swartz) Stearn leaves.

PubMed

Tan, Joash Ban Lee; Lim, Yau Yan; Lee, Sui Mae

2015-04-01

The decoction and infusion of Rhoeo spathacea (Swartz) Stearn leaves have been recognized as a functional food particularly in South America, but has not yet gained international popularity as a beverage. The primary aim of this study was to establish the viability of R. spathacea aqueous leaf extracts as a beverage, in terms of its antioxidant activity and antibacterial activity. The antioxidant contents of aqueous and methanol leaf extracts were evaluated by the total phenolic content (TPC) and total flavonoid content (TFC) assays. The antioxidant activities measured were DPPH radical scavenging activity (FRS), ferric reducing power (FRP) and ferrous ion chelating (FIC) activity. The aqueous leaf extracts in the forms of decoction and infusion, were found to have comparable TPC and antioxidant activity with other herbal teas previously reported by our research group. Both decoction and infusion also exhibited antibacterial activity against six species of Gram positive and four species of Gram negative bacteria, notably methicillin-resistant Staphylococcus aureus and Neisseria gonorrhoeae. A total of four different known phenolic compounds were identified by HPLC and MS, three of which have not been previously reported to be found in this plant. Both the decoction and infusion of the leaves R. spathacea have potential to be popularized into a common beverage.

Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) Activates Global and Heterogeneous Local Ca2+ Signals from NAADP- and Ryanodine Receptor-gated Ca2+ Stores in Pulmonary Arterial Myocytes*

PubMed Central

Jiang, Yong-Liang; Lin, Amanda H. Y.; Xia, Yang; Lee, Suengwon; Paudel, Omkar; Sun, Hui; Yang, Xiao-Ru; Ran, Pixin; Sham, James S. K.

2013-01-01

Nicotinic acid adenine dinucleotide phosphate (NAADP) is the most potent Ca2+-mobilizing messenger that releases Ca2+ from endolysosomal organelles. Recent studies showed that NAADP-induced Ca2+ release is mediated by the two-pore channels (TPCs) TPC1 and TPC2. However, the expression of TPCs and the NAADP-induced local Ca2+ signals have not been examined in vascular smooth muscle. Here, we found that both TPC1 and TPC2 are expressed in rat pulmonary arterial smooth muscle cells (PASMCs), with TPC1 being the major subtype. Application of membrane-permeant NAADP acetoxymethyl ester to PASMCs elicited a biphasic increase in global [Ca2+]i, which was independent of extracellular Ca2+ and blocked by the NAADP antagonist Ned-19 or the vacuolar H+-ATPase inhibitor bafilomycin A1, indicating Ca2+ release from acidic endolysosomal Ca2+ stores. The Ca2+ response was unaffected by xestospongin C but was partially blocked by ryanodine or thapsigargin. NAADP triggered heterogeneous local Ca2+ signals, including a diffuse increase in cytosolic [Ca2+], Ca2+ sparks, Ca2+ bursts, and regenerative Ca2+ release. The diffuse Ca2+ increase and Ca2+ bursts were ryanodine-insensitive, presumably arising from different endolysosomal sources. Ca2+ sparks and regenerative Ca2+ release were inhibited by ryanodine, consistent with cross-activation of loosely coupled ryanodine receptors. Moreover, Ca2+ release stimulated by endothelin-1 was inhibited by Ned-19, ryanodine, or xestospongin C, suggesting that NAADP-mediated Ca2+ signals interact with both ryanodine and inositol 1,4,5-trisphosphate receptors during agonist stimulation. Our results show that NAADP mediates complex global and local Ca2+ signals. Depending on the physiological stimuli, these diverse Ca2+ signals may serve to regulate different cellular functions in PASMCs. PMID:23443655

Intra-molecular Charge Transfer and Electron Delocalization in Non-fullerene Organic Solar Cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu, Qinghe; Zhao, Donglin; Goldey, Matthew B.

Two types of electron acceptors were synthesized by coupling two kinds of electron-rich cores with four equivalent perylene diimides (PDIs) at the a position. With fully aromatic cores, TPB and TPSe have pi-orbitals spread continuously over the whole aromatic conjugated backbone, unlike TPC and TPSi, which contain isolated PDI units due to the use of a tetrahedron carbon or silicon linker. Density functional theory calculations of the projected density of states showed that the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) for TPB are localized in separate regions of space. Further, the LUMO of TPB showsmore » a greater contribution from the orbitals belonging to the connective core of the molecules than that of TPC. Overall, the properties of the HOMO and LUMO point at increased intra-molecular delocalization of negative charge carriers for TPB and TPSe than for TPC and TPSi and hence at a more facile intra-molecular charge transfer for the former. The film absorption and emission spectra showed evidences for the inter -molecular electron delocalization in TPB and TPSe, which is consistent with the network structure revealed by X-ray diffraction studies on single crystals of TPB. These features benefit the formation of charge transfer states and/or facilitate charge transport. Thus, higher electron mobility and higher charge dissociation probabilities under J(sc) condition were observed in blend films of TPB:PTB7-Th and TPSe:PTB7-Th than those in TPC:PTB7Th and TPSi:PTB7-Th blend films. As a result, the J(sc) and fill factor values of 15.02 mA/cm(2), 0.58 and 14.36 mA/cm(2), 0.55 for TPB- and TPSe-based solar cell are observed, whereas those for TPC and TPSi are 11.55 mA/cm2, 0.47 and 10.35 mA/cm(2), 0.42, respectively.« less

A ROS-Assisted Calcium Wave Dependent on the AtRBOHD NADPH Oxidase and TPC1 Cation Channel Propagates the Systemic Response to Salt Stress.

PubMed

Evans, Matthew J; Choi, Won-Gyu; Gilroy, Simon; Morris, Richard J

2016-07-01

Plants exhibit rapid, systemic signaling systems that allow them to coordinate physiological and developmental responses throughout the plant body, even to highly localized and quickly changing environmental stresses. The propagation of these signals is thought to include processes ranging from electrical and hydraulic networks to waves of reactive oxygen species (ROS) and cytoplasmic Ca(2+) traveling throughout the plant. For the Ca(2+) wave system, the involvement of the vacuolar ion channel TWO PORE CHANNEL1 (TPC1) has been reported. However, the precise role of this channel and the mechanism of cell-to-cell propagation of the wave have remained largely undefined. Here, we use the fire-diffuse-fire model to analyze the behavior of a Ca(2+) wave originating from Ca(2+) release involving the TPC1 channel in Arabidopsis (Arabidopsis thaliana). We conclude that a Ca(2+) diffusion-dominated calcium-induced calcium-release mechanism is insufficient to explain the observed wave transmission speeds. The addition of a ROS-triggered element, however, is able to quantitatively reproduce the observed transmission characteristics. The treatment of roots with the ROS scavenger ascorbate and the NADPH oxidase inhibitor diphenyliodonium and analysis of Ca(2+) wave propagation in the Arabidopsis respiratory burst oxidase homolog D (AtrbohD) knockout background all led to reductions in Ca(2+) wave transmission speeds consistent with this model. Furthermore, imaging of extracellular ROS production revealed a systemic spread of ROS release that is dependent on both AtRBOHD and TPC1 These results suggest that, in the root, plant systemic signaling is supported by a ROS-assisted calcium-induced calcium-release mechanism intimately involving ROS production by AtRBOHD and Ca(2+) release dependent on the vacuolar channel TPC1. © 2016 American Society of Plant Biologists. All Rights Reserved.

A New Acoustic Portal into the Odontocete Ear and Vibrational Analysis of the Tympanoperiotic Complex

PubMed Central

Cranford, Ted W.; Krysl, Petr; Amundin, Mats

2010-01-01

Global concern over the possible deleterious effects of noise on marine organisms was catalyzed when toothed whales stranded and died in the presence of high intensity sound. The lack of knowledge about mechanisms of hearing in toothed whales prompted our group to study the anatomy and build a finite element model to simulate sound reception in odontocetes. The primary auditory pathway in toothed whales is an evolutionary novelty, compensating for the impedance mismatch experienced by whale ancestors as they moved from hearing in air to hearing in water. The mechanism by which high-frequency vibrations pass from the low density fats of the lower jaw into the dense bones of the auditory apparatus is a key to understanding odontocete hearing. Here we identify a new acoustic portal into the ear complex, the tympanoperiotic complex (TPC) and a plausible mechanism by which sound is transduced into the bony components. We reveal the intact anatomic geometry using CT scanning, and test functional preconceptions using finite element modeling and vibrational analysis. We show that the mandibular fat bodies bifurcate posteriorly, attaching to the TPC in two distinct locations. The smaller branch is an inconspicuous, previously undescribed channel, a cone-shaped fat body that fits into a thin-walled bony funnel just anterior to the sigmoid process of the TPC. The TPC also contains regions of thin translucent bone that define zones of differential flexibility, enabling the TPC to bend in response to sound pressure, thus providing a mechanism for vibrations to pass through the ossicular chain. The techniques used to discover the new acoustic portal in toothed whales, provide a means to decipher auditory filtering, beam formation, impedance matching, and transduction. These tools can also be used to address concerns about the potential deleterious effects of high-intensity sound in a broad spectrum of marine organisms, from whales to fish. PMID:20694149

A new acoustic portal into the odontocete ear and vibrational analysis of the tympanoperiotic complex.

PubMed

Cranford, Ted W; Krysl, Petr; Amundin, Mats

2010-08-04

Global concern over the possible deleterious effects of noise on marine organisms was catalyzed when toothed whales stranded and died in the presence of high intensity sound. The lack of knowledge about mechanisms of hearing in toothed whales prompted our group to study the anatomy and build a finite element model to simulate sound reception in odontocetes. The primary auditory pathway in toothed whales is an evolutionary novelty, compensating for the impedance mismatch experienced by whale ancestors as they moved from hearing in air to hearing in water. The mechanism by which high-frequency vibrations pass from the low density fats of the lower jaw into the dense bones of the auditory apparatus is a key to understanding odontocete hearing. Here we identify a new acoustic portal into the ear complex, the tympanoperiotic complex (TPC) and a plausible mechanism by which sound is transduced into the bony components. We reveal the intact anatomic geometry using CT scanning, and test functional preconceptions using finite element modeling and vibrational analysis. We show that the mandibular fat bodies bifurcate posteriorly, attaching to the TPC in two distinct locations. The smaller branch is an inconspicuous, previously undescribed channel, a cone-shaped fat body that fits into a thin-walled bony funnel just anterior to the sigmoid process of the TPC. The TPC also contains regions of thin translucent bone that define zones of differential flexibility, enabling the TPC to bend in response to sound pressure, thus providing a mechanism for vibrations to pass through the ossicular chain. The techniques used to discover the new acoustic portal in toothed whales, provide a means to decipher auditory filtering, beam formation, impedance matching, and transduction. These tools can also be used to address concerns about the potential deleterious effects of high-intensity sound in a broad spectrum of marine organisms, from whales to fish.

NEXT, a HPXe TPC for neutrinoless double beta decay searches

NASA Astrophysics Data System (ADS)

Gómez-Cadenas, J. J.; Martín-Albo, J.

2008-11-01

The next-generation experiments for neutrinoless double beta decay searches are a major challenge, since the detectors have to fulfill conflicting requirements: they have to be very massive and scalable; very sensitive to the signature of the ββ processes, and simultaneously impose extremely stringent limits on the copious backgrounds. A high-pressure gaseous Xenon TPC may be the ideal detector for this purpose: the isotope 136Xe is almost 9% of natural Xenon and enrichment by centrifugation is a relatively easy technology as demonstrated by the EXO collaboration; the detector can be extrapolated to large masses; energy resolution in gas is expected to be at the level of 1% FWHM at Qββ this fact, combined with the very long life of the ββ2ν mode accounts for negligible backgrounds of intrinsic origin up to masses of the order of 1 ton. The detector is fully active and the gas can be continuously re-circulated and purified, which, together with the existence of a kinematical signature gives an extra handle against backgrounds. NEXT (Neutrino Experiment with a Xenon TPC) is an on-going project that aims to build a 100 kg HPXe TPC at a pressure of ~10 bar. The experiment has already been aproved by the scientific committee of the Canfranc Underground Laboratory (LSC), in Spain, for the initial R&D phase. The NEXT Collaboration has defined an ambitious roadmap for the next five years: an initial phase of R&D during 2 years will be used to decide the technology (MicroMegas, GEMs. APDs. PMTs, etc.); in 3 years a first prototype (NEXT-10) will be constructed; followed by NEXT-100 in 5 years. A special grant (Consolider-Ingenio 2010) from the Ministry of Science of Spain will fund the experiment during this period. The Collaboration is composed by the leading Spanish groups in experimental neutrino and underground physics. The general ideas behind neutrinoless double beta decay searches with a HPXe TPC are discussed in this note.

DarkSide-20k: A 20 tonne two-phase LAr TPC for direct dark matter detection at LNGS

NASA Astrophysics Data System (ADS)

Aalseth, C. E.; Acerbi, F.; Agnes, P.; Albuquerque, I. F. M.; Alexander, T.; Alici, A.; Alton, A. K.; Antonioli, P.; Arcelli, S.; Ardito, R.; Arnquist, I. J.; Asner, D. M.; Ave, M.; Back, H. O.; Barrado Olmedo, A. I.; Batignani, G.; Bertoldo, E.; Bettarini, S.; Bisogni, M. G.; Bocci, V.; Bondar, A.; Bonfini, G.; Bonivento, W.; Bossa, M.; Bottino, B.; Boulay, M.; Bunker, R.; Bussino, S.; Buzulutskov, A.; Cadeddu, M.; Cadoni, M.; Caminata, A.; Canci, N.; Candela, A.; Cantini, C.; Caravati, M.; Cariello, M.; Carlini, M.; Carpinelli, M.; Castellani, A.; Catalanotti, S.; Cataudella, V.; Cavalcante, P.; Cavuoti, S.; Cereseto, R.; Chepurnov, A.; Cicalò, C.; Cifarelli, L.; Citterio, M.; Cocco, A. G.; Colocci, M.; Corgiolu, S.; Covone, G.; Crivelli, P.; D'Antone, I.; D'Incecco, M.; D'Urso, D.; Da Rocha Rolo, M. D.; Daniel, M.; Davini, S.; de Candia, A.; De Cecco, S.; De Deo, M.; De Filippis, G.; De Guido, G.; De Rosa, G.; Dellacasa, G.; Della Valle, M.; Demontis, P.; Derbin, A.; Devoto, A.; Di Eusanio, F.; Di Pietro, G.; Dionisi, C.; Dolgov, A.; Dormia, I.; Dussoni, S.; Empl, A.; Fernandez Diaz, M.; Ferri, A.; Filip, C.; Fiorillo, G.; Fomenko, K.; Franco, D.; Froudakis, G. E.; Gabriele, F.; Gabrieli, A.; Galbiati, C.; Garcia Abia, P.; Gendotti, A.; Ghisi, A.; Giagu, S.; Giampa, P.; Gibertoni, G.; Giganti, C.; Giorgi, M. A.; Giovanetti, G. K.; Gligan, M. L.; Gola, A.; Gorchakov, O.; Goretti, A. M.; Granato, F.; Grassi, M.; Grate, J. W.; Grigoriev, G. Y.; Gromov, M.; Guan, M.; Guerra, M. B. B.; Guerzoni, M.; Gulino, M.; Haaland, R. K.; Hallin, A.; Harrop, B.; Hoppe, E. W.; Horikawa, S.; Hosseini, B.; Hughes, D.; Humble, P.; Hungerford, E. V.; Ianni, An.; Jillings, C.; Johnson, T. N.; Keeter, K.; Kendziora, C. L.; Kim, S.; Koh, G.; Korablev, D.; Korga, G.; Kubankin, A.; Kuss, M.; Kuźniak, M.; La Commara, M.; Lehnert, B.; Li, X.; Lissia, M.; Lodi, G. U.; Loer, B.; Longo, G.; Loverre, P.; Lussana, R.; Luzzi, L.; Ma, Y.; Machado, A. A.; Machulin, I. N.; Mandarano, A.; Mapelli, L.; Marcante, M.; Margotti, A.; Mari, S. M.; Mariani, M.; Maricic, J.; Martoff, C. J.; Mascia, M.; Mayer, M.; McDonald, A. B.; Messina, A.; Meyers, P. D.; Milincic, R.; Moggi, A.; Moioli, S.; Monroe, J.; Monte, A.; Morrocchi, M.; Mount, B. J.; Mu, W.; Muratova, V. N.; Murphy, S.; Musico, P.; Nania, R.; Navrer Agasson, A.; Nikulin, I.; Nosov, V.; Nozdrina, A. O.; Nurakhov, N. N.; Oleinik, A.; Oleynikov, V.; Orsini, M.; Ortica, F.; Pagani, L.; Pallavicini, M.; Palmas, S.; Pandola, L.; Pantic, E.; Paoloni, E.; Paternoster, G.; Pavletcov, V.; Pazzona, F.; Peeters, S.; Pelczar, K.; Pellegrini, L. A.; Pelliccia, N.; Perotti, F.; Perruzza, R.; Pesudo, V.; Piemonte, C.; Pilo, F.; Pocar, A.; Pollmann, T.; Portaluppi, D.; Pugachev, D. A.; Qian, H.; Radics, B.; Raffaelli, F.; Ragusa, F.; Razeti, M.; Razeto, A.; Regazzoni, V.; Regenfus, C.; Reinhold, B.; Renshaw, A. L.; Rescigno, M.; Retière, F.; Riffard, Q.; Rivetti, A.; Rizzardini, S.; Romani, A.; Romero, L.; Rossi, B.; Rossi, N.; Rubbia, A.; Sablone, D.; Salatino, P.; Samoylov, O.; Sánchez García, E.; Sands, W.; Sanfilippo, S.; Sant, M.; Santorelli, R.; Savarese, C.; Scapparone, E.; Schlitzer, B.; Scioli, G.; Segreto, E.; Seifert, A.; Semenov, D. A.; Shchagin, A.; Shekhtman, L.; Shemyakina, E.; Sheshukov, A.; Simeone, M.; Singh, P. N.; Skensved, P.; Skorokhvatov, M. D.; Smirnov, O.; Sobrero, G.; Sokolov, A.; Sotnikov, A.; Speziale, F.; Stainforth, R.; Stanford, C.; Suffritti, G. B.; Suvorov, Y.; Tartaglia, R.; Testera, G.; Tonazzo, A.; Tosi, A.; Trinchese, P.; Unzhakov, E. V.; Vacca, A.; Vázquez-Jáuregui, E.; Verducci, M.; Viant, T.; Villa, F.; Vishneva, A.; Vogelaar, B.; Wada, M.; Wahl, J.; Walding, J.; Wang, H.; Wang, Y.; Watson, A. W.; Westerdale, S.; Williams, R.; Wojcik, M. M.; Wu, S.; Xiang, X.; Xiao, X.; Yang, C.; Ye, Z.; Yllera de Llano, A.; Zappa, F.; Zappalà, G.; Zhu, C.; Zichichi, A.; Zullo, M.; Zullo, A.; Zuzel, G.

2018-03-01

Building on the successful experience in operating the DarkSide-50 detector, the DarkSide Collaboration is going to construct DarkSide-20k, a direct WIMP search detector using a two-phase Liquid Argon Time Projection Chamber (LAr TPC) with an active (fiducial) mass of 23 t (20 t). This paper describes a preliminary design for the experiment, in which the DarkSide-20k LAr TPC is deployed within a shield/veto with a spherical Liquid Scintillator Veto (LSV) inside a cylindrical Water Cherenkov Veto (WCV). This preliminary design provides a baseline for the experiment to achieve its physics goals, while further development work will lead to the final optimization of the detector parameters and an eventual technical design. Operation of DarkSide-50 demonstrated a major reduction in the dominant 39Ar background when using argon extracted from an underground source, before applying pulse shape analysis. Data from DarkSide-50, in combination with MC simulation and analytical modeling, shows that a rejection factor for discrimination between electron and nuclear recoils of >3 × 109 is achievable. This, along with the use of the veto system and utilizing silicon photomultipliers in the LAr TPC, are the keys to unlocking the path to large LAr TPC detector masses, while maintaining an experiment in which less than < 0.1 events (other than ν-induced nuclear recoils) is expected to occur within the WIMP search region during the planned exposure. DarkSide-20k will have ultra-low backgrounds than can be measured in situ, giving sensitivity to WIMP-nucleon cross sections of 1.2 × 10^{-47} cm2 (1.1 × 10^{-46} cm2) for WIMPs of 1 TeV/c 2 (10 TeV/c 2) mass, to be achieved during a 5 yr run producing an exposure of 100 t yr free from any instrumental background.

Differential expression of two scribble isoforms during Drosophila embryogenesis.

PubMed

Li, M; Marhold, J; Gatos, A; Török, I; Mechler, B M

2001-10-01

The tumour suppressor gene scribble (scrib) is required for epithelial polarity and growth control in Drosophila. Here, we report the identification and embryonic expression pattern of two Scrib protein isoforms resulting from alternative splicing during scrib transcription. Both proteins are first ubiquitously expressed during early embryogenesis. Then, during morphogenesis each Scrib protein displays a specific pattern of expression in the central and peripheral nervous systems, CNS and PNS, respectively. During germ band extension, the expression of the longer form Scrib1 occurs predominantly in the neuroblasts derived from the neuro-ectoderm and becomes later restricted to CNS neurones as well as to the pole cells in the gonads. By contrast, the shorter form Scrib2 is strongly expressed in the PNS and a subset of CNS neurones.

Recent Results from ArgoNeuT and Status of MicroBooNE

DOE Office of Scientific and Technical Information (OSTI.GOV)

Szelc, Andrzej

2015-07-10

Liquid Argon Time Projection Chamber (LArTPC) detectors hold the key to answering the outstanding questions about the role of neutrinos in the Standard Model of Particle physics and beyond. Their fine granularity combined with calorimetric capabilities allows for precision measurements that answering these questions will require. Here, we discuss the recent results from the ArgoNeuT experiment as well as the status and prospects for MicroBooNE, both a part of the US-based LArTPC neutrino program.

Multiple isoforms for the catalytic subunit of PKA in the basal fungal lineage Mucor circinelloides.

PubMed

Fernández Núñez, Lucas; Ocampo, Josefina; Gottlieb, Alexandra M; Rossi, Silvia; Moreno, Silvia

2016-12-01

Protein kinase A (PKA) activity is involved in dimorphism of the basal fungal lineage Mucor. From the recently sequenced genome of Mucor circinelloides we could predict ten catalytic subunits of PKA. From sequence alignment and structural prediction we conclude that the catalytic core of the isoforms is conserved, and the difference between them resides in their amino termini. This high number of isoforms is maintained in the subdivision Mucoromycotina. Each paralogue, when compared to the ones form other fungi is more homologous to one of its orthologs than to its paralogs. All of these fungal isoforms cannot be included in the class I or II in which fungal protein kinases have been classified. mRNA levels for each isoform were measured during aerobic and anaerobic growth. The expression of each isoform is differential and associated to a particular growth stage. We reanalyzed the sequence of PKAC (GI 20218944), the only cloned sequence available until now for a catalytic subunit of M. circinelloides. PKAC cannot be classified as a PKA because of its difference in the conserved C-tail; it shares with PKB a conserved C2 domain in the N-terminus. No catalytic activity could be measured for this protein nor predicted bioinformatically. It can thus be classified as a pseudokinase. Its importance can not be underestimated since it is expressed at the mRNA level in different stages of growth, and its deletion is lethal. Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Myosin isoform switching during assembly of the Drosophila flight muscle thick filament lattice.

PubMed

Orfanos, Zacharias; Sparrow, John C

2013-01-01

During muscle development myosin molecules form symmetrical thick filaments, which integrate with the thin filaments to produce the regular sarcomeric lattice. In Drosophila indirect flight muscles (IFMs) the details of this process can be studied using genetic approaches. The weeP26 transgenic line has a GFP-encoding exon inserted into the single Drosophila muscle myosin heavy chain gene, Mhc. The weeP26 IFM sarcomeres have a unique MHC-GFP-labelling pattern restricted to the sarcomere core, explained by non-translation of the GFP exon following alternative splicing. Characterisation of wild-type IFM MHC mRNA confirmed the presence of an alternately spliced isoform, expressed earlier than the major IFM-specific isoform. The two wild-type IFM-specific MHC isoforms differ by the presence of a C-terminal 'tailpiece' in the minor isoform. The sequential expression and assembly of these two MHCs into developing thick filaments suggest a role for the tailpiece in initiating A-band formation. The restriction of the MHC-GFP sarcomeric pattern in weeP26 is lifted when the IFM lack the IFM-specific myosin binding protein flightin, suggesting that it limits myosin dissociation from thick filaments. Studies of flightin binding to developing thick filaments reveal a progressive binding at the growing thick filament tips and in a retrograde direction to earlier assembled, proximal filament regions. We propose that this flightin binding restricts myosin molecule incorporation/dissociation during thick filament assembly and explains the location of the early MHC isoform pattern in the IFM A-band.

Discrete forms of amylose are synthesized by isoforms of GBSSI in pea.

PubMed

Edwards, Anne; Vincken, Jean-Paul; Suurs, Luc C J M; Visser, Richard G F; Zeeman, Sam; Smith, Alison; Martin, Cathie

2002-08-01

Amyloses with distinct molecular masses are found in the starch of pea embryos compared with the starch of pea leaves. In pea embryos, a granule-bound starch synthase protein (GBSSIa) is required for the synthesis of a significant portion of the amylose. However, this protein seems to be insignificant in the synthesis of amylose in pea leaves. cDNA clones encoding a second isoform of GBSSI, GBSSIb, have been isolated from pea leaves. Comparison of GBSSIa and GBSSIb activities shows them to have distinct properties. These differences have been confirmed by the expression of GBSSIa and GBSSIb in the amylose-free mutant of potato. GBSSIa and GBSSIb make distinct forms of amylose that differ in their molecular mass. These differences in product specificity, coupled with differences in the tissues in which GBSSIa and GBSSIb are most active, explain the distinct forms of amylose found in different tissues of pea. The shorter form of amylose formed by GBSSIa confers less susceptibility to the retrogradation of starch pastes than the amylose formed by GBSSIb. The product specificity of GBSSIa could provide beneficial attributes to starches for food and nonfood uses.

Discrete Forms of Amylose Are Synthesized by Isoforms of GBSSI in PeaW⃞

PubMed Central

Edwards, Anne; Vincken, Jean-Paul; Suurs, Luc C. J. M.; Visser, Richard G. F.; Zeeman, Sam; Smith, Alison; Martin, Cathie

2002-01-01

Amyloses with distinct molecular masses are found in the starch of pea embryos compared with the starch of pea leaves. In pea embryos, a granule-bound starch synthase protein (GBSSIa) is required for the synthesis of a significant portion of the amylose. However, this protein seems to be insignificant in the synthesis of amylose in pea leaves. cDNA clones encoding a second isoform of GBSSI, GBSSIb, have been isolated from pea leaves. Comparison of GBSSIa and GBSSIb activities shows them to have distinct properties. These differences have been confirmed by the expression of GBSSIa and GBSSIb in the amylose-free mutant of potato. GBSSIa and GBSSIb make distinct forms of amylose that differ in their molecular mass. These differences in product specificity, coupled with differences in the tissues in which GBSSIa and GBSSIb are most active, explain the distinct forms of amylose found in different tissues of pea. The shorter form of amylose formed by GBSSIa confers less susceptibility to the retrogradation of starch pastes than the amylose formed by GBSSIb. The product specificity of GBSSIa could provide beneficial attributes to starches for food and nonfood uses. PMID:12172021

Immunologic differentiation of two high-affinity neurotensin receptor isoforms in the developing rat brain.

PubMed

Boudin, H; Lazaroff, B; Bachelet, C M; Pélaprat, D; Rostène, W; Beaudet, A

2000-09-11

Earlier studies have demonstrated overexpression of NT1 neurotensin receptors in rat brain during the first 2 weeks of life. To gain insight into this phenomenon, we investigated the identity and distribution of NT1 receptor proteins in the brain of 10-day-old rats by using two different NT1 antibodies: one (Abi3) directed against the third intracellular loop and the other (Abi4) against the C-terminus of the receptor. Immunoblot experiments that used Abi3 revealed the presence of two differentially glycosylated forms of the NT1 receptor in developing rat brain: one migrating at 54 and the other at 52 kDa. Whereas the 54-kDa form was expressed from birth to adulthood, the 52-kDa form was detected only at 10 and 15 days postnatal. Only the 52-kDa isoform was recognized by Abi4. By immunohistochemistry, both forms of the receptor were found to be predominantly expressed in cerebral cortex and dorsal hippocampus, in keeping with earlier radioligand binding and in situ hybridization data. However, whereas Abi4 immunoreactivity was mainly concentrated within nerve cell bodies and extensively colocalized with the Golgi marker alpha-mannosidase II, Abi3 immunoreactivity was predominantly located along neuronal processes. These results suggest that the transitorily expressed 52-kDa protein corresponds to an immature, incompletely glycosylated and largely intracellular form of the NT1 receptor and that the 54-kDa protein corresponds to a mature, fully glycosylated, and largely membrane-associated form. They also indicate that antibodies directed against different sequences of G-protein-coupled receptors may yield isoform-specific immunohistochemical labeling patterns in mammalian brain. Finally, the selective expression of the short form of the NT1 receptor early in development suggests that it may play a specific role in the establishment of neuronal circuitry. Copyright 2000 Wiley-Liss, Inc.

Effects of ocean acidification on pelagic carbon fluxes in a mesocosm experiment

NASA Astrophysics Data System (ADS)

Spilling, Kristian; Schulz, Kai G.; Paul, Allanah J.; Boxhammer, Tim; Achterberg, Eric P.; Hornick, Thomas; Lischka, Silke; Stuhr, Annegret; Bermúdez, Rafael; Czerny, Jan; Crawfurd, Kate; Brussaard, Corina P. D.; Grossart, Hans-Peter; Riebesell, Ulf

2016-11-01

About a quarter of anthropogenic CO2 emissions are currently taken up by the oceans, decreasing seawater pH. We performed a mesocosm experiment in the Baltic Sea in order to investigate the consequences of increasing CO2 levels on pelagic carbon fluxes. A gradient of different CO2 scenarios, ranging from ambient ( ˜ 370 µatm) to high ( ˜ 1200 µatm), were set up in mesocosm bags ( ˜ 55 m3). We determined standing stocks and temporal changes of total particulate carbon (TPC), dissolved organic carbon (DOC), dissolved inorganic carbon (DIC), and particulate organic carbon (POC) of specific plankton groups. We also measured carbon flux via CO2 exchange with the atmosphere and sedimentation (export), and biological rate measurements of primary production, bacterial production, and total respiration. The experiment lasted for 44 days and was divided into three different phases (I: t0-t16; II: t17-t30; III: t31-t43). Pools of TPC, DOC, and DIC were approximately 420, 7200, and 25 200 mmol C m-2 at the start of the experiment, and the initial CO2 additions increased the DIC pool by ˜ 7 % in the highest CO2 treatment. Overall, there was a decrease in TPC and increase of DOC over the course of the experiment. The decrease in TPC was lower, and increase in DOC higher, in treatments with added CO2. During phase I the estimated gross primary production (GPP) was ˜ 100 mmol C m-2 day-1, from which 75-95 % was respired, ˜ 1 % ended up in the TPC (including export), and 5-25 % was added to the DOC pool. During phase II, the respiration loss increased to ˜ 100 % of GPP at the ambient CO2 concentration, whereas respiration was lower (85-95 % of GPP) in the highest CO2 treatment. Bacterial production was ˜ 30 % lower, on average, at the highest CO2 concentration than in the controls during phases II and III. This resulted in a higher accumulation of DOC and lower reduction in the TPC pool in the elevated CO2 treatments at the end of phase II extending throughout phase III. The "extra" organic carbon at high CO2 remained fixed in an increasing biomass of small-sized plankton and in the DOC pool, and did not transfer into large, sinking aggregates. Our results revealed a clear effect of increasing CO2 on the carbon budget and mineralization, in particular under nutrient limited conditions. Lower carbon loss processes (respiration and bacterial remineralization) at elevated CO2 levels resulted in higher TPC and DOC pools than ambient CO2 concentration. These results highlight the importance of addressing not only net changes in carbon standing stocks but also carbon fluxes and budgets to better disentangle the effects of ocean acidification.

Liquid Argon TPC Signal Formation, Signal Processing and Hit Reconstruction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baller, Bruce

2017-03-11

This document describes the early stage of the reconstruction chain that was developed for the ArgoNeuT and MicroBooNE experiments at Fermilab. These experiments study accelerator neutrino interactions that occur in a Liquid Argon Time Projection Chamber. Reconstructing the properties of particles produced in these interactions requires knowledge of the micro-physics processes that affect the creation and transport of ionization electrons to the readout system. A wire signal deconvolution technique was developed to convert wire signals to a standard form for hit reconstruction, to remove artifacts in the electronics chain and to remove coherent noise.

Metabolic characterization of a mouse deficient in all known leptin receptor isoforms.

PubMed

Osborn, Olivia; Sanchez-Alavez, Manuel; Brownell, Sara E; Ross, Brendon; Klaus, Joe; Dubins, Jeffrey; Beutler, Bruce; Conti, Bruno; Bartfai, Tamas

2010-01-01

We have characterized a newly generated mouse model of obesity, a mouse strain deficient in all five previously described leptin receptor isoforms. These transgenic mice, named the db (333)/db (333) mice, were identified from an ENU mutagenesis screen and carry a point mutation in the seventh exon of the db gene encoding the leptin receptor, resulting in a premature stop codon (Y(333)Stop) and gene product that lacks STAT signaling domains. db (333)/db (333) mice have a morbidly obese phenotype, with body weights diverging from wild type as early as 4 weeks of age (P < 0.05). To determine the contribution of the short isoforms of the leptin receptor in this metabolic phenotype, we performed an extensive metabolic characterization of the db (333)/db (333) mouse in relation to the well-characterized db/db mouse lacking only the long form of the leptin receptor. db (333)/db (333) mice have similar endocrine and metabolic parameters as previously described in other leptin receptor transgenic mice including db/db mice that lack only the long isoform of the leptin receptor. However, db (333)/db (333) mice show a subtle trend toward higher body weight and insulin levels, lower oxygen, carbon dioxide production, respiratory exchange ratio (RER), and temperature than db/db mice suggesting the short isoforms may play an additional role in energy homeostasis.

Identification of dynamin as a septin-binding protein.

PubMed

Maimaitiyiming, Maowulan; Kobayashi, Yuumi; Kumanogoh, Haruko; Nakamura, Shun; Morita, Mitsuhiro; Maekawa, Shohei

2013-02-08

Lipid rafts (detergent-resistant low-density membrane microdomain: DRM) are signal-transducing membrane platforms. In a previous study, we showed maturation-dependent localization of septin in the DRM fraction of rat brain. Mammalian septin is composed with 13-14 isoforms and these isoforms assemble to form rod-shaped hetero-oligomeric complexes. End-to-end polymerization of these complexes results in the formation of higher order structures such as filamentous sheets or bundles of filaments that restrict the fluid-like diffusion of the membrane proteins and lipids. Considering the function of septin as the membrane scaffold, elucidation of the molecular interaction of septin in DRM could be a breakthrough to understand another role of lipid rafts. In order to identify septin-binding proteins in DRM, solubilization and fractionation of septin from DRM was attempted. Several proteins were co-fractionated with septin and LC-MS/MS analysis identified one of these proteins as dynamin and Western blotting using anti-dynamin confirmed this result. Immunoprecipitation of septin11 in a crude supernatant showed co-precipitation of dynamin and dynamin fraction prepared from brain contained several septin isoforms. Within bacterially expressed septin isoforms, septin5 and septin11 bound dynamin but septin9 did not. These results suggest that some septin isoforms participate in the dynamin-related membrane dynamics. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Characterization of Native Protein Complexes and Protein Isoform Variation Using Size-fractionation-based Quantitative Proteomics*

PubMed Central

Kirkwood, Kathryn J.; Ahmad, Yasmeen; Larance, Mark; Lamond, Angus I.

2013-01-01

Proteins form a diverse array of complexes that mediate cellular function and regulation. A largely unexplored feature of such protein complexes is the selective participation of specific protein isoforms and/or post-translationally modified forms. In this study, we combined native size-exclusion chromatography (SEC) with high-throughput proteomic analysis to characterize soluble protein complexes isolated from human osteosarcoma (U2OS) cells. Using this approach, we have identified over 71,500 peptides and 1,600 phosphosites, corresponding to over 8,000 proteins, distributed across 40 SEC fractions. This represents >50% of the predicted U2OS cell proteome, identified with a mean peptide sequence coverage of 27% per protein. Three biological replicates were performed, allowing statistical evaluation of the data and demonstrating a high degree of reproducibility in the SEC fractionation procedure. Specific proteins were detected interacting with multiple independent complexes, as typified by the separation of distinct complexes for the MRFAP1-MORF4L1-MRGBP interaction network. The data also revealed protein isoforms and post-translational modifications that selectively associated with distinct subsets of protein complexes. Surprisingly, there was clear enrichment for specific Gene Ontology terms associated with differential size classes of protein complexes. This study demonstrates that combined SEC/MS analysis can be used for the system-wide annotation of protein complexes and to predict potential isoform-specific interactions. All of these SEC data on the native separation of protein complexes have been integrated within the Encyclopedia of Proteome Dynamics, an online, multidimensional data-sharing resource available to the community. PMID:24043423

Characterization of native protein complexes and protein isoform variation using size-fractionation-based quantitative proteomics.

PubMed

Kirkwood, Kathryn J; Ahmad, Yasmeen; Larance, Mark; Lamond, Angus I

2013-12-01

Proteins form a diverse array of complexes that mediate cellular function and regulation. A largely unexplored feature of such protein complexes is the selective participation of specific protein isoforms and/or post-translationally modified forms. In this study, we combined native size-exclusion chromatography (SEC) with high-throughput proteomic analysis to characterize soluble protein complexes isolated from human osteosarcoma (U2OS) cells. Using this approach, we have identified over 71,500 peptides and 1,600 phosphosites, corresponding to over 8,000 proteins, distributed across 40 SEC fractions. This represents >50% of the predicted U2OS cell proteome, identified with a mean peptide sequence coverage of 27% per protein. Three biological replicates were performed, allowing statistical evaluation of the data and demonstrating a high degree of reproducibility in the SEC fractionation procedure. Specific proteins were detected interacting with multiple independent complexes, as typified by the separation of distinct complexes for the MRFAP1-MORF4L1-MRGBP interaction network. The data also revealed protein isoforms and post-translational modifications that selectively associated with distinct subsets of protein complexes. Surprisingly, there was clear enrichment for specific Gene Ontology terms associated with differential size classes of protein complexes. This study demonstrates that combined SEC/MS analysis can be used for the system-wide annotation of protein complexes and to predict potential isoform-specific interactions. All of these SEC data on the native separation of protein complexes have been integrated within the Encyclopedia of Proteome Dynamics, an online, multidimensional data-sharing resource available to the community.

DOE Office of Scientific and Technical Information (OSTI.GOV)

De Geronimo, G.; Li, S.; D'Andragora, A.

We present a front-end application-specific integrated circuit (ASIC) for a wire based time-projection-chamber (TPC) operating in liquid Argon (LAr). The LAr TPC will be used for long baseline neutrino oscillation experiments. The ASIC must provide a low-noise readout of the signals induced on the TPC wires, digitization of those signals at 2 MSamples/s, compression, buffering and multiplexing. A resolution of better than 1000 rms electrons at 200 pF input capacitance for an input range of 300 fC is required, along with low power and operation in LAr (at 87 K). We include the characterization of a commercial technology for operationmore » in the cryogenic environment and the first experimental results on the analog front end. The results demonstrate that complementary metal-oxide semiconductor transistors have lower noise and much improved dc characteristics at LAr temperature. Finally, we introduce the concept of '1/f equivalent' to model the low-frequency component of the noise spectral density, for use in the input metal-oxide semiconductor field-effect transistor optimization.« less

Influence of variety and harvest maturity on phytochemical content in corn silk.

PubMed

Sarepoua, Eakrin; Tangwongchai, Ratchada; Suriharn, Bhalang; Lertrat, Kamol

2015-02-15

Corn silk has been used as a traditional herb in Asia. The objective of this study was to evaluate variability in phytochemicals in corn varieties at three maturity stages of corn silk. Ten vegetable corn varieties were evaluated in a completely randomized design with three replications. Data were recorded for total phenolic (TPC), total flavonoids (TFC), total anthocyanin (TAC) and antioxidant activity (AA) by DPPH free-radical-scavenging assays. Differences among corn varieties were observed for all parameters at all maturity stages, and the interactions between maturity stage and corn variety were significant. TPC and TAC were highest at the milky stage, whereas TFC and AA were highest at the silking stage. TPC, TFC and AA were highest in super sweet corn and white corn at the silking stage. PWC5 variety of purple waxy corn at the milky stage had the highest values for all parameters, and it is useful for further development of functional food products. Copyright © 2014 Elsevier Ltd. All rights reserved.

Study of a Large Prototype TPC for the ILC using Micro-Pattern Gas Detectors

NASA Astrophysics Data System (ADS)

Münnich, A.; LCTPC Collaboration

2016-04-01

In the last decade, R&D for detectors for the future International Linear Collider (ILC) has been performed by the community. The International Large Detector (ILD) is one of two detector concepts at the ILC. Its tracking system consists of a Si vertex detector, forward tracking disks and a large volume Time Projection Chamber (TPC). Within the LCTPC collaboration, a Large Prototype (LP) TPC has been built as a demonstrator. Its endplate is able to house up to seven identical modules with Micro-Pattern Gas Detectors (MPGD) amplification. Recently, the LP has been equipped with resistive anode Micromegas (MM) or Gas Electron Multiplier (GEM) modules. Both the MM and GEM technologies have been studied with an electron beam up to 6 GeV in a 1 Tesla solenoid magnet. After introducing the current R&D status, recent results will be presented including field distortions, ion gating and spatial resolution as well as future plans of the LCTPC R&D.

Measuring the Neutron Cross Section and Detector Response from Interactions in Liquid Argon

NASA Astrophysics Data System (ADS)

Kamp, Nicholas; Collaboration, Captain

2017-09-01

The main objective of the CAPTAIN (Cryogenic Apparatus for Precision Tests of Argon Interactions with Neutrinos) program is to measure neutron and neutrino interactions in liquid argon. These results will be essential to the development of both short and long baseline neutrino experiments. The full CAPTAIN experiment involves a 10 ton liquid argon time projection chamber (LArTPC) that will take runs at a low-energy ( 10-50 MeV) stopped pion neutrino source. A two ton LArTPC, MiniCAPTAIN, will serve as a prototype for the full CAPTAIN detector. MiniCAPTAIN has been deployed to take data at the Los Alamos Neutron Science Center in late July. During this run, it will both test new LArTPC technologies and measure the cross section and detector response of neutron interactions in liquid argon. The results will be helpful in characterizing neutral current neutrino interactions and identifying background in future neutrino detection experiments. This poster gives an overview of these results and a status update on the CAPTAIN collaboration.

Live event reconstruction in an optically read out GEM-based TPC

NASA Astrophysics Data System (ADS)

Brunbauer, F. M.; Galgóczi, G.; Gonzalez Diaz, D.; Oliveri, E.; Resnati, F.; Ropelewski, L.; Streli, C.; Thuiner, P.; van Stenis, M.

2018-04-01

Combining strong signal amplification made possible by Gaseous Electron Multipliers (GEMs) with the high spatial resolution provided by optical readout, highly performing radiation detectors can be realized. An optically read out GEM-based Time Projection Chamber (TPC) is presented. The device permits 3D track reconstruction by combining the 2D projections obtained with a CCD camera with timing information from a photomultiplier tube. Owing to the intuitive 2D representation of the tracks in the images and to automated control, data acquisition and event reconstruction algorithms, the optically read out TPC permits live display of reconstructed tracks in three dimensions. An Ar/CF4 (80/20%) gas mixture was used to maximize scintillation yield in the visible wavelength region matching the quantum efficiency of the camera. The device is integrated in a UHV-grade vessel allowing for precise control of the gas composition and purity. Long term studies in sealed mode operation revealed a minor decrease in the scintillation light intensity.

Phenolic composition and antioxidant properties of koose, a deep-fat fried cowpea cake.

PubMed

Apea-Bah, Franklin B; Serem, June C; Bester, Megan J; Duodu, Kwaku G

2017-12-15

Koose, a West African delicacy, is a side dish prepared by deep frying thick cowpea paste. The current research determined the effect of deep-fat frying of cowpea paste on its total phenolic content (TPC), phenolic composition and antioxidant properties. Four cowpea cultivars comprising two reddish-brown, a brownish-cream and cream phenotypes were used. Liquid chromatography-mass spectrometry was used to determine phenolic composition of the samples. TPC was determined using Folin-Ciocalteu method while radical scavenging capacities were by Trolox equivalent antioxidant capacity, oxygen radical absorbance capacity and nitric oxide scavenging assays. The phenolic acids identified included benzoic and cinnamic acid derivatives. The predominant flavonoid classes were flavan-3-ols and flavonols. Deep-fat frying of the cowpea pastes decreased their TPC, radical scavenging capacities and total quantified flavonoids. The koose inhibited radical-induced oxidative cellular and DNA damage. It is concluded that koose is a potential functional food that can contribute to alleviating radical-induced oxidative stress. Copyright © 2017 Elsevier Ltd. All rights reserved.

Over-seasons analysis of quantitative trait loci affecting phenolic content and antioxidant capacity in raspberry.

PubMed

Dobson, Patricia; Graham, Julie; Stewart, D; Brennan, Rex; Hackett, Christine A; McDougall, Gordon J

2012-05-30

This study examined the total phenol content (TPC) and total anthocyanin content (TAC) in ripe fruit of progeny of a mapping population generated from a cross between the European red raspberry cv. Glen Moy ( Rubus ideaus var. idaeus) and the North American red raspberry cv. Latham ( Rubus ideaus var. strigosus) over five seasons in two different growing environments. Measurements of antioxidant capacity (FRAP and TEAC) were also carried out. TPC was highly correlated with TEAC and FRAP across the entire data set. The subset of anthocyanin content was genotype-dependent but also correlated with TPC, although the proportion of anthocyanin compounds varied between progeny. Quantitative trait locus (QTL) analysis was carried out, and key markers were tested for consistency of effects over sites and years. Four regions, on linkage groups 2, 3, 5, and 6, were identified. These agree with QTLs from a previous study over a single season and indicate that QTL effects were robust over seasons.

Physics with gamma-beams and charged particle detectors: I) Nuclear structure II) Nuclear astrophysics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gai, Moshe

The Charged Particle Working Group (CPWG) is proposing to construct large area Silicon Strip Detector (SSD), a gas Time Projection Chamber detector read by an electronic readout system (eTPC) and a Bubble Chamber (BC) containing superheated high purity water to be used in measurements utilizing intense gamma-ray beams from the newly constructed ELI-NP facility at Magurele, Bucharest in Romania. We intend to use the SSD and eTPC detectors to address essential problems in nuclear structure physics, such as clustering and the many alpha-decay of light nuclei such as {sup 12}C and {sup 16}O. All three detectors (SSD, eTPC and BC)more » will be used to address central problems in nuclear astrophysics such as the astrophysical cross section factor of the {sup 12}C(α,γ) reaction and other processes central to stellar evolution. The CPWG intends to submit to the ELI-NP facility a Technical Design Report (TDR) for the proposed detectors.« less

Ionization Electron Signal Processing in Single Phase LArTPCs I. Algorithm Description and Quantitative Evaluation with MicroBooNE Simulation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Adams, C.; et al.

We describe the concept and procedure of drifted-charge extraction developed in the MicroBooNE experiment, a single-phase liquid argon time projection chamber (LArTPC). This technique converts the raw digitized TPC waveform to the number of ionization electrons passing through a wire plane at a given time. A robust recovery of the number of ionization electrons from both induction and collection anode wire planes will augment the 3D reconstruction, and is particularly important for tomographic reconstruction algorithms. A number of building blocks of the overall procedure are described. The performance of the signal processing is quantitatively evaluated by comparing extracted charge withmore » the true charge through a detailed TPC detector simulation taking into account position-dependent induced current inside a single wire region and across multiple wires. Some areas for further improvement of the performance of the charge extraction procedure are also discussed.« less

Development of pressurised hot water extraction (PHWE) for essential compounds from Moringa oleifera leaf extracts.

PubMed

Matshediso, Phatsimo G; Cukrowska, Ewa; Chimuka, Luke

2015-04-01

Pressurised hot water extraction (PHWE) is a "green" technology which can be used for the extraction of essential components in Moringa oleifera leaf extracts. The behaviour of three flavonols (myricetin, quercetin and kaempferol) and total phenolic content (TPC) in Moringa leaf powder were investigated at various temperatures using PHWE. The TPC of extracts from PHWE were investigated using two indicators. These are reducing activity and the radical scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH). Flavonols content in the PHWE extracts were analysed on high performance liquid chromatography with ultra violet (HPLC-UV) detection. The concentration of kaempferol and myricetin started decreasing at 150 °C while that of quercetin remained steady with extraction temperature. Optimum extraction temperature for flavonols and DPPH radical scavenging activity was found to be 100 °C. The TPC increased with temperature until 150 °C and then decreased while the reducing activity increased. Copyright © 2014 Elsevier Ltd. All rights reserved.

Antioxidant capacity of cornelian cherry (Cornus mas L.) - comparison between permanganate reducing antioxidant capacity and other antioxidant methods.

PubMed

Popović, Boris M; Stajner, Dubravka; Slavko, Kevrešan; Sandra, Bijelić

2012-09-15

Ethanol extracts (80% in water) of 10 cornelian cherry (Cornus mas L.) genotypes were studied for antioxidant properties, using methods including DPPH(), ()NO, O(2)(-) and ()OH antiradical powers, FRAP, total phenolic and anthocyanin content (TPC and ACC) and also one relatively new, permanganate method (permanganate reducing antioxidant capacity-PRAC). Lipid peroxidation (LP) was also determined as an indicator of oxidative stress. The data from different procedures were compared and analysed by multivariate techniques (correlation matrix calculation and principal component analysis (PCA)). Significant positive correlations were obtained between TPC, ACC and DPPH(), ()NO, O(2)(-), and ()OH antiradical powers, and also between PRAC and TPC, ACC and FRAP. PCA found two major clusters of cornelian cherry, based on antiradical power, FRAP and PRAC and also on chemical composition. Chemometric evaluation showed close interdependence between PRAC method and FRAP and ACC. There was a huge variation between C. mas genotypes in terms of antioxidant activity. Copyright © 2012 Elsevier Ltd. All rights reserved.

Comparative antioxidant activity of edible Japanese brown seaweeds.

PubMed

Airanthi, M K Widjaja-Adhi; Hosokawa, Masashi; Miyashita, Kazuo

2011-01-01

Japanese edible brown seaweeds, Eisenia bicyclis (Arame), Kjellmaniella crassifolia (Gagome), Alaria crassifolia (Chigaiso), Sargassum horneri (Akamoku), and Cystoseira hakodatensis (Uganomoku) were assayed for total phenolic content (TPC), fucoxanthin content, radical scavenging activities (DPPH, peroxyl radical, ABTS, and nitric oxide), and antioxidant activity in a liposome system. Among the solvents used for extraction, methanol was the most effective to extract total phenolics (TPC) from brown seaweeds. Among 5 kinds of brown seaweeds analyzed, methanol extract from C. hakodatensis was the best source for antioxidants. The high antioxidant activity of the extract was based not only on the high content of phenolics, but on the presence of fucoxanthin. No significant correlation (P > 0.05) was observed between TPC per gram extract with DPPH radical scavenging activity of the methanol extracts. These observed discrepancy would be due to structural variations in the phenolic compounds, and different levels of fucoxanthin in the extracts. The present study also demonstrated the synergy in the antioxidant activity of the combination of brown seaweed phenolics and fucoxanthin.

Analytical optimization of a phenolic-rich herbal extract and supplementation in fermented milk containing sweet potato pulp.

PubMed

Ramos, Lorena Rodrigues; Santos, Jânio Sousa; Daguer, Heitor; Valese, Andressa Camargo; Cruz, Adriano Gomes; Granato, Daniel

2017-04-15

The aims of the present study were to optimize and characterize the phenolic composition of a herbal extract composed of green mate (Ilex paraguariensis), clove (Syzygium aromaticum), and lemongrass (Cymbopogon citratus) and to propose the addition of this polyphenol-rich extract to fermented milks (FM) with/without sweet potato pulp (Ipomoea batatas). Proximate composition, pH, acidity, instrumental texture profile, total phenolic content (TPC), antioxidant activity (AA) of all formulations were measured, and sensory attributes were also investigated. The addition of a lyophilized extract (1g 100g -1 ) containing 87.5% clove and 12.5% green mate increased the AA and TPC, while FM with added sweet potato pulp had the best sensory acceptance. The TPC and total reducing capacity had a slight change during 21days of storage. The data showed that herbal extracts and sweet potato pulp may be used to develop new dairy foods with potential functional properties. Copyright © 2016 Elsevier Ltd. All rights reserved.

Profiling of Phenolic Compounds and Antioxidant Activity of 12 Cruciferous Vegetables.

PubMed

Li, Zhifeng; Lee, Hui Wen; Liang, Xu; Liang, Dong; Wang, Qi; Huang, Dejian; Ong, Choon Nam

2018-05-10

The phenolic profiles of 12 cruciferous vegetables (pakchoi, choysum, Chinese cabbage, kailan, Brussels sprout, cabbage, cauliflower, broccoli, rocket salad, red cherry radish, daikon radish, and watercress) were studied with UHPLC-MS/MS. Antioxidant activity and total phenolic content (TPC) were also evaluated. A total of 74 phenolic compounds were identified, including 16 hydroxycinnamic acids and derivatives, and 58 flavonoids and derivatives. The main flavonoids identified were glycosylated quercetin, kaempferol and isorhamnetin, and the main hydroxycinnamic acids were ferulic, sinapic, caffeic and p -coumaric acids. Principal component analysis (PCA) revealed that the distribution of phenolic compounds in different genera of cruciferous vegetables was in accordance with their conventional taxonomy. The DPPH, ORAC and TPC values ranged from 1.11 to 9.54 µmoles Trolox equivalent/g FW, 5.34 to 32.92 µmoles Trolox equivalent/g FW, and 0.16 to 1.93 mg gallic acid equivalent/g FW respectively. Spearman’s correlation showed significant ( p < 0.05) positive correlations between TPC, flavonoids and antioxidant activity.

Exclusive muon neutrino charged current pion-less topologies. ArgoNeuT results and future prospects in LAr TPC detectors

DOE PAGES

Palamara, Ornella

2016-12-29

Results from the analysis of charged current pion-less (CC 0-pion) muon neutrino events in argon collected by the ArgoNeuT experiment on the NuMI beam at Fermilab are presented and compared with predictions from Monte Carlo simulations. A novel analysis method, based on the reconstruction of exclusive topologies, fully exploiting the Liquid argon Time Projection Chamber (LAr TPC) technique capabilities, is used to analyze the events, characterized by the presence at the vertex of a leading muon track eventually accompanied by one or more highly ionizing tracks, and study nuclear effects in neutrino interactions on argon nuclei. Multiple protons accompanying themore » leading muon are visible in the ArgoNeuT events, and measured with a proton reconstruction threshold of 21 MeV kinetic energy. As a result, measurements of (anti-)neutrino CC 0-pion inclusive and exclusive cross sections on argon nuclei are reported. Prospects for future, larger mass LAr TPC detectors are discussed.« less

Bound phenolic compounds and antioxidant properties of whole grain and bran of white, red and black rice.

PubMed

Pang, Yuehan; Ahmed, Sulaiman; Xu, Yanjie; Beta, Trust; Zhu, Zhiwei; Shao, Yafang; Bao, Jinsong

2018-02-01

Total phenolic content (TPC), individual phenolic acid and antioxidant capacity of whole grain and bran fraction 18 rices with different bran color were investigated. The levels of TPC in bound fractions were significantly higher than those in the free fractions either in the whole grains or brans. The main bound phenolic acids in white rice samples were ferulic acid, p-coumaric acid, and isoferulic acid, and in pigmented rice samples were ferulic acid, p-coumaric acid, and vanillic acid. The protocatechuic acid and 2,5-dihydroxybenzoic acid were not detected in white samples. The content of gallic acid, protocatechuic acid, 2,5-dihydroxybenzoic acid, ferulic acid, sinapic acid had significantly positive correlations with TPC and antioxidant capacity. This study found much wider diversity in the phenolics and antioxidant capacity in the whole grain and brans of rice, and will provide new opportunities to further improvement of rice with enhanced levels of the phytochemicals. Copyright © 2017 Elsevier Ltd. All rights reserved.

The effect of ultraviolet treatment on enzymatic activity and total phenolic content of minimally processed potato slices.

PubMed

Teoh, Li Shing; Lasekan, Ola; Adzahan, Noranizan Mohd; Hashim, Norhashila

2016-07-01

In this work, potato slices were exposed to different doses of UV-C irradiation (i.e. 2.28, 6.84, 11.41, and 13.68 kJ m -2 ) with or without pretreatment [i.e. ascorbic acid and calcium chloride (AACCl) dip] and stored at 4 ± 1 °C. Changes in enzymatic activities of polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase (PAL), as well as total phenolic content (TPC) were investigated after 0, 3, 7 and 10 days of storage. Results showed that untreated and UV-C treated potato slices at 13.68 kJ m -2 dosage level showed significantly higher PPO, POD and PAL activities. Conversely, untreated potato slices showed the lowest TPC during storage period. Potato slices subjected to AACCl dip plus UV-C at 6.84 kJ m -2 produced lower PPO, POD and PAL activities, as well as maintained a high TPC during storage.

Numerical Study of Electrostatic Field Distortion on LPTPC End-Plates based on Bulk Micromegas Modules

NASA Astrophysics Data System (ADS)

Bhattacharya, Purba; Bhattacharya, Deb Sankar; Mukhopadhyay, Supratik; Majumdar, Nayana; Bhattacharya, Sudeb; Colas, Paul; Attié, David

2018-02-01

The R&D activities for the linear collider TPC (LC-TPC) are currently working on the adoption of the micro pattern devices for the gaseous amplification stage. Several beam tests have been carried out at DESY with a 5 GeV electron beam in a 1 T superconducting magnet. We worked on a large prototype TPC with an end-plate that was built, for the first time, using seven resistive bulk Micromegas modules. During experiments, reduced signal sensitivity was observed at the boundary of these modules. Electrostatic field distortion near the module boundaries was considered to be the possible major reason behind these observations. In the present work, we will explore this hypothesis through numerical simulation. Our aim has been to understand the origin of distortions observed close to the edges of the test beam modules and to explore the possibility of using the Garfield simulation framework for investigating a phenomenon as complex as distortion.

Drying effects on the antioxidant properties of tomatoes and ginger.

PubMed

Gümüşay, Özlem Aktürk; Borazan, Alev Akpınar; Ercal, Nuran; Demirkol, Omca

2015-04-15

In this study, the effects of four different drying processes, sun drying (SD), oven drying (OD), vacuum oven drying (VOD) and freeze drying (FD) for tomatoes (Solanum lycopersicum) and ginger (Zingiber officinale) in terms of thiolic and phenolic contents have been studied. Thiol content, total phenolic content (TPC), ascorbic acid (AA) content, and cupric ion reducing antioxidant capacity (CUPRAC) were determined in fresh and dried samples. Glutathione (GSH) and cysteine (Cys) were determined as the thiol contents of tomatoes and ginger. Significant losses were observed in the contents of TPC, AA, GSH and Cys and CUPRAC values in all samples that were dried using the thermal method. There was a statistically significant difference in the losses of the TPC, AA, and thiol contents between the use of thermal drying and freeze drying (except Cys in tomatoes) methods. Freeze dried tomato and ginger samples have been found to have better antioxidant properties. Copyright © 2014 Elsevier Ltd. All rights reserved.

Antioxidant and DNA damage protective properties of anthocyanin-rich extracts from Hibiscus and Ocimum: a comparative study.

PubMed

Sarkar, Biswatrish; Kumar, Dhananjay; Sasmal, Dinakar; Mukhopadhyay, Kunal

2014-01-01

Anthocyanin extracts (AEs) from Ocimum tenuiflorum (leaf), Hibiscus rosa-sinensis (petal) and Hibiscus sabdariffa (calyx) were investigated and compared for in vitro antioxidant activity and DNA damage protective property. Total phenolic content (TPC) and total anthocyanin content (TAC) of the AEs were determined and the major anthocyanins were characterised. In vitro antioxidant activities were assessed by ferric-reducing antioxidant power (FRAP) assay, 2,2-diphenyl-1-picryl hydrazyl (DPPH) radical-scavenging activity, 2-deoxy-D-ribose degradation assay and lipid peroxidation assay. The protective property of the AEs was also examined against oxidative DNA damage by H2O2 and UV using pUC19 plasmid. All the AEs particularly those from O. tenuiflorum demonstrated efficient antioxidant activity and protected DNA from damage. Strong correlation between antioxidant capacity and TPC and TAC was observed. Significant correlation between antioxidant capacity and TPC and TAC ascertained that phenolics and anthocyanins were the major contributors of antioxidant activity.

Accelerated Stability Studies on Dried Extracts of Centella asiatica Through Chemical, HPLC, HPTLC, and Biological Activity Analyses.

PubMed

Kaur, Ishtdeep; Suthar, Nancy; Kaur, Jasmeen; Bansal, Yogita; Bansal, Gulshan

2016-10-01

Regulatory guidelines recommend systematic stability studies on a herbal product to establish its shelf life. In the present study, commercial extracts (Types I and II) and freshly prepared extract (Type III) of Centella asiatica were subjected to accelerated stability testing for 6 months. Control and stability samples were evaluated for organoleptics, pH, moisture, total phenolic content (TPC), asiatic acid, kaempherol, and high-performance thin layer chromatography fingerprints, and for antioxidant and acetylcholinesterase inhibitory activities. Markers and TPC and both the activities of each extract decreased in stability samples with respect to control. These losses were maximum in Type I extract and minimum in Type III extract. Higher stability of Type III extract than others might be attributed to the additional phytoconstituents and/or preservatives in it. Pearson correlation analysis of the results suggested that TPC, asiatic acid, and kaempferol can be taken as chemical markers to assess chemical and therapeutic shelf lives of herbal products containing Centella asiatica. © The Author(s) 2016.

Activity of selected hydrolytic enzymes in Allium sativum L. anthers.

PubMed

Winiarczyk, Krystyna; Gębura, Joanna

2016-05-01

The aim of the study was to determine enzymatic activity in sterile Allium sativum anthers in the final stages of male gametophyte development (the stages of tetrads and free microspores). The analysed enzymes were shown to occur in the form of numerous isoforms. In the tetrad stage, esterase activity was predominant, which was manifested by the greater number of isoforms of the enzyme. In turn, in the microspore stage, higher numbers of isoforms of acid phosphatases and proteases were detected. The development of sterile pollen grains in garlic is associated with a high level of protease and acid phosphatase activity and lower level of esterase activities in the anther locule. Probably this is the first description of the enzymes activity (ACPH, EST, PRO) in the consecutives stages of cell wall formation which is considered to be one of the causes of male sterility in flowering plant. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Neuronal glucose transporter isoform 3 deficient mice demonstrate features of autism spectrum disorders.

PubMed

Zhao, Y; Fung, C; Shin, D; Shin, B-C; Thamotharan, S; Sankar, R; Ehninger, D; Silva, A; Devaskar, S U

2010-03-01

Neuronal glucose transporter (GLUT) isoform 3 deficiency in null heterozygous mice led to abnormal spatial learning and working memory but normal acquisition and retrieval during contextual conditioning, abnormal cognitive flexibility with intact gross motor ability, electroencephalographic seizures, perturbed social behavior with reduced vocalization and stereotypies at low frequency. This phenotypic expression is unique as it combines the neurobehavioral with the epileptiform characteristics of autism spectrum disorders. This clinical presentation occurred despite metabolic adaptations consisting of an increase in microvascular/glial GLUT1, neuronal GLUT8 and monocarboxylate transporter isoform 2 concentrations, with minimal to no change in brain glucose uptake but an increase in lactate uptake. Neuron-specific glucose deficiency has a negative impact on neurodevelopment interfering with functional competence. This is the first description of GLUT3 deficiency that forms a possible novel genetic mechanism for pervasive developmental disorders, such as the neuropsychiatric autism spectrum disorders, requiring further investigation in humans.

Michel electron reconstruction using cosmic-ray data from the MicroBooNE LArTPC

NASA Astrophysics Data System (ADS)

Acciarri, R.; Adams, C.; An, R.; Anthony, J.; Asaadi, J.; Auger, M.; Bagby, L.; Balasubramanian, S.; Baller, B.; Barnes, C.; Barr, G.; Bass, M.; Bay, F.; Bishai, M.; Blake, A.; Bolton, T.; Bugel, L.; Camilleri, L.; Caratelli, D.; Carls, B.; Castillo Fernandez, R.; Cavanna, F.; Chen, H.; Church, E.; Cianci, D.; Cohen, E.; Collin, G. H.; Conrad, J. M.; Convery, M.; Crespo-Anadón, J. I.; Del Tutto, M.; Devitt, D.; Dytman, S.; Eberly, B.; Ereditato, A.; Escudero Sanchez, L.; Esquivel, J.; Fleming, B. T.; Foreman, W.; Furmanski, A. P.; Garcia-Gamez, D.; Garvey, G. T.; Genty, V.; Goeldi, D.; Gollapinni, S.; Graf, N.; Gramellini, E.; Greenlee, H.; Grosso, R.; Guenette, R.; Hackenburg, A.; Hamilton, P.; Hen, O.; Hewes, J.; Hill, C.; Ho, J.; Horton-Smith, G.; Huang, E.-C.; James, C.; de Vries, J. Jan; Jen, C.-M.; Jiang, L.; Johnson, R. A.; Joshi, J.; Jostlein, H.; Kaleko, D.; Karagiorgi, G.; Ketchum, W.; Kirby, B.; Kirby, M.; Kobilarcik, T.; Kreslo, I.; Laube, A.; Li, Y.; Lister, A.; Littlejohn, B. R.; Lockwitz, S.; Lorca, D.; Louis, W. C.; Luethi, M.; Lundberg, B.; Luo, X.; Marchionni, A.; Mariani, C.; Marshall, J.; Martinez Caicedo, D. A.; Meddage, V.; Miceli, T.; Mills, G. B.; Moon, J.; Mooney, M.; Moore, C. D.; Mousseau, J.; Murrells, R.; Naples, D.; Nienaber, P.; Nowak, J.; Palamara, O.; Paolone, V.; Papavassiliou, V.; Pate, S. F.; Pavlovic, Z.; Piasetzky, E.; Porzio, D.; Pulliam, G.; Qian, X.; Raaf, J. L.; Rafique, A.; Rochester, L.; von Rohr, C. Rudolf; Russell, B.; Schmitz, D. W.; Schukraft, A.; Seligman, W.; Shaevitz, M. H.; Sinclair, J.; Snider, E. L.; Soderberg, M.; Söldner-Rembold, S.; Soleti, S. R.; Spentzouris, P.; Spitz, J.; St. John, J.; Strauss, T.; Sutton, K. A.; Szelc, A. M.; Tagg, N.; Terao, K.; Thomson, M.; Toups, M.; Tsai, Y.-T.; Tufanli, S.; Usher, T.; Van de Water, R. G.; Viren, B.; Weber, M.; Wickremasinghe, D. A.; Wolbers, S.; Wongjirad, T.; Woodruff, K.; Yang, T.; Yates, L.; Zeller, G. P.; Zennamo, J.; Zhang, C.

2017-09-01

The MicroBooNE liquid argon time projection chamber (LArTPC) has been taking data at Fermilab since 2015 collecting, in addition to neutrino beam, cosmic-ray muons. Results are presented on the reconstruction of Michel electrons produced by the decay at rest of cosmic-ray muons. Michel electrons are abundantly produced in the TPC, and given their well known energy spectrum can be used to study MicroBooNE's detector response to low-energy electrons (electrons with energies up to ~ 50 MeV). We describe the fully-automated algorithm developed to reconstruct Michel electrons, with which a sample of ~ 14,000 Michel electron candidates is obtained. Most of this article is dedicated to studying the impact of radiative photons produced by Michel electrons on the accuracy and resolution of their energy measurement. In this energy range, ionization and bremsstrahlung photon production contribute similarly to electron energy loss in argon, leading to a complex electron topology in the TPC. By profiling the performance of the reconstruction algorithm on simulation we show that the ability to identify and include energy deposited by radiative photons leads to a significant improvement in the energy measurement of low-energy electrons. The fractional energy resolution we measure improves from over 30% to ~ 20% when we attempt to include radiative photons in the reconstruction. These studies are relevant to a large number of analyses which aim to study neutrinos by measuring electrons produced by νe interactions over a broad energy range.

Adrenaline Stimulates Glucagon Secretion by Tpc2-Dependent Ca2+ Mobilization From Acidic Stores in Pancreatic α-Cells.

PubMed

Hamilton, Alexander; Zhang, Quan; Salehi, Albert; Willems, Mara; Knudsen, Jakob G; Ringgaard, Anna K; Chapman, Caroline E; Gonzalez-Alvarez, Alejandro; Surdo, Nicoletta C; Zaccolo, Manuela; Basco, Davide; Johnson, Paul R V; Ramracheya, Reshma; Rutter, Guy A; Galione, Antony; Rorsman, Patrik; Tarasov, Andrei I

2018-06-01

Adrenaline is a powerful stimulus of glucagon secretion. It acts by activation of β-adrenergic receptors, but the downstream mechanisms have only been partially elucidated. Here, we have examined the effects of adrenaline in mouse and human α-cells by a combination of electrophysiology, imaging of Ca 2+ and PKA activity, and hormone release measurements. We found that stimulation of glucagon secretion correlated with a PKA- and EPAC2-dependent (inhibited by PKI and ESI-05, respectively) elevation of [Ca 2+ ] i in α-cells, which occurred without stimulation of electrical activity and persisted in the absence of extracellular Ca 2+ but was sensitive to ryanodine, bafilomycin, and thapsigargin. Adrenaline also increased [Ca 2+ ] i in α-cells in human islets. Genetic or pharmacological inhibition of the Tpc2 channel (that mediates Ca 2+ release from acidic intracellular stores) abolished the stimulatory effect of adrenaline on glucagon secretion and reduced the elevation of [Ca 2+ ] i Furthermore, in Tpc2-deficient islets, ryanodine exerted no additive inhibitory effect. These data suggest that β-adrenergic stimulation of glucagon secretion is controlled by a hierarchy of [Ca 2+ ] i signaling in the α-cell that is initiated by cAMP-induced Tpc2-dependent Ca 2+ release from the acidic stores and further amplified by Ca 2+ -induced Ca 2+ release from the sarco/endoplasmic reticulum. © 2018 by the American Diabetes Association.

Antioxidant activity of commonly consumed cereals, millets, pulses and legumes in India.

PubMed

Sreeramulu, D; Reddy, C Vijaya Kumar; Raghunath, M

2009-02-01

Plant foods are important due to their antioxidant activity (AOA) attributed to the phenolics which are known to protect organisms against harmful effects of oxygen radicals. However, information on antioxidant activity of Indian plant foods is scanty. Therefore, the present study evaluated the AOA of cereals, millets, pulses and legumes, commonly consumed in India and assessed the relationship with their total phenolic content (TPC). AOA was assessed by DPPH (2,2-Diphenyl-1-picryl hydrazyl) radical scavenging assay, ferric reducing antioxidant power (FRAP) assay and reducing power. DPPH scavenging activity ranged from 0.24 and 1.73 mg/g, whereas FRAP ranged from 16.21 to 471.71 micromoles/g. Finger millet (Eleusine cora cana) and Rajmah (Phaseolus vulgaris) had the highest FRAP 471.71, 372.76 and DPPH scavenging activity 1.73, 1.07. Similar trends were observed with reducing power. Among cereals and legumes, Finger millet (Ragi) and black gram dhal (Phaseolus mungo Roxb) had the highest TPC, the values being 373 and 418 mg/100 g respectively, while rice (Oryza sativa) and green gram dhal (Phaseolus aureus Roxb) showed the least (47.6 and 62.4 mg/100 g). In the present study, FRAP (r = 0.91) and reducing power (r = 0.90) showed significant correlation with TPC in cereals and millets, but not in pulses and legumes. The results suggest that TPC contributes significantly to the AOA of Indian cereals and millets.

Michel Electron Reconstruction Using Cosmic-Ray Data from the MicroBooNE LArTPC

DOE PAGES

Acciarri, R.

2017-09-14

The MicroBooNE liquid argon time projection chamber (LArTPC) has been taking data at Fermilab since 2015 collecting, in addition to neutrino beam, cosmic-ray muons. Results are presented on the reconstruction of Michel electrons produced by the decay at rest of cosmic-ray muons. Michel electrons are abundantly produced in the TPC, and given their well known energy spectrum can be used to study MicroBooNE's detector response to low-energy electrons (electrons with energies up to ~50 MeV). We describe the fully-automated algorithm developed to reconstruct Michel electrons, with which a sample of ~14,000 Michel electron candidates is obtained. Most of this article is dedicated to studying the impact of radiative photons produced by Michel electrons on the accuracy and resolution of their energy measurement. In this energy range, ionization and bremsstrahlung photon production contribute similarly to electron energy loss in argon, leading to a complex electron topology in the TPC. By profiling the performance of the reconstruction algorithm on simulation we show that the ability to identify and include energy deposited by radiative photons leads to a significant improvement in the energy measurement of low-energy electrons. The fractional energy resolution we measure improves from over 30% to ~20% when we attempt to include radiative photons in the reconstruction. These studies are relevant to a large number of analyses which aim to study neutrinos by measuring electrons produced bymore » $$\

Role of the Prospect Rock Fault in the Exhumation of High Pressure Rocks in North-Central Vermont

NASA Astrophysics Data System (ADS)

Tam, E.; Webb, L. E.; Aiken, C. L.

2017-12-01

The Prospect Rock Fault (PRF) is key to our interpretation of regional deformation and exhumation of blueschist and eclogite-facies rocks in the Tillotson Peak Complex (TPC) during the Taconic Orogeny. The TPC is in the footwall of the PRF in the eastern limb of the Green Mountain Anticlinorium. In the TPC, the dominant foliation is S2 and E-W trending F2 folds parallel L2 lineations, which run orthogonal to regional N-S trending folds associated with the Taconic Orogeny. This structural trend has possible analogies with HP-UHP terranes in Papua New Guinea and China. The PRF itself is folded by F2 folds. Presently, there is a lack of consensus about the structural evolution of the PRF and its role in the exhumation of the TPC, and studies have not reconciled the formation of the E-W folds and lineations to a regional model. Oriented samples and structural data were collected from the footwall of the PRF over several transects. Samples were processed into orthogonal thin sections for microstructural analyses and for 40Ar/39Ar step-heating of white mica. Preliminary results show a range of ages from 458.6 ± 2.0 Ma to 420.0 ± 2.7 Ma. The oldest ages are just slightly younger, yet concordant, with published and new 40Ar/39Ar ages from the TPC. The dominant foliation in the PRF samples, S2, is defined in thin section by mica and quartz microlithons, and oriented mica grains. S1 is only locally preserved in some mica domains and albite/garnet inclusion trails. S3 appears as crenulations of S2, with no significant new mineral crystallization. In the field, L2 lineations are defined by mineral and quartz rods, and L3 lineations are defined as intersection lineations on S2 surfaces. The relationships between ages and microstructures are consistent younger ages being associated with increased presence of S3 crenulation foliations, which appeared in structurally lower areas. Samples with older ages display dominant S2 foliations and lack S3 crenulations, and were sourced from structurally higher areas. Our results suggest the PRF played a role in exhumation of the TPC and ages obtained are closely aligned with deformation ages constrained from 40Ar/39Ar dating in southern Quebec for the Taconic D2 and Salinian D3 deformation. These dates may aid to further correlation tectonostratographic models between southern Quebec and northern Vermont.

Bonner Prize Talk -- First Laboratory Observation of Double Beta Decay

NASA Astrophysics Data System (ADS)

Moe, Michael

2013-04-01

Although we are awash in neutrinos, we remain ignorant of some of their fundamental properties. We don't know their masses. We don't know whether ``anti-neutrinos'' are really distinct particles. Double beta (ββ) decay offers a handle on these questions if we can observe the energy spectrum of the two emitted electrons, and determine whether or not they share their energy with two neutrinos. Seeing neutrinoless (0ν) decay would solve some enduring puzzles. The power of the process to elucidate the neutrino was recognized in the 1930's, but ββ decay would be exceedingly rare and difficult to detect. Unsuccessful laboratory searches had been going on for 25 years when the UC Irvine group began its first experiment with a cloud chamber in 1972. After some background for the non-expert, and a snapshot of the theoretical and experimental milieu at the time, the talk will begin with the reasons for choosing a cloud chamber, and the taming of its balky and idiosyncratic behavior. The talk will end with the first definitive observation of two-neutrino (2ν)ββ decay of ^82Se in the vastly superior time projection chamber (TPC) in 1987. Discouragement through the tortuous 15-year interval was relieved by occasional victories. Some I will illustrate with revealing cloud-chamber photographs. We learned many things from this primitive device, and after seven years we isolated an apparent ββ decay signal. But the efficiency of the trigger was small, and difficult to pin down. Estimating 2.2%, we were way low. The resulting ``short'' ^82Se half-life of 1 x 10^19 years was suspect. New technology came to the rescue with the invention of the TPC. Experience with the cloud chamber guided our design of a TPC specifically for ββ decay. The TPC was built from scratch. Its long, steep learning curve was also punctuated with little triumphs. A memorable moment was the first turn-on of a portion of the chamber. So long ago, this all seems rather quaint, but through ample use of photographs and anecdotes it makes and interesting story. As a digital device, the TPC made data acquisition and analysis orders of magnitude simpler and faster. After seven years of massage, the TPC yielded good evidence for 2ν decay of ^82Se with a half-life near 10^20 years. While the 0ν mode was not in evidence, finally seeing ββ decay in the laboratory created optimism about an eventual 0ν discovery.

Effect of roasting degree on the antioxidant activity of different Arabica coffee quality classes.

PubMed

Odžaković, Božana; Džinić, Natalija; Kukrić, Zoran; Grujić, Slavica

2016-01-01

Coffee is one of the most widely consumed beverages in the world, because of its unique sensory properties and physiological properties. Coffee beverages represent a significant source of antioxidants in the consumers' diet and contribute significantly to their daily intake. The aim of this research was to investigate the effect of different roasting degrees on the content of biologically active compounds and antioxidant activity in different quality classes of Arabica coffee. Samples of green Arabica coffee (Rio Minas) of two quality classes from two production batches were used for the research. Roasting was carried out at temperatures of 167, 175 and 171°C. The total phenol content (TPC), total flavonoid content (TFC), flavonol content (FC) and antioxidant activity (DPPH, ABTS) in the coffee extracts was determined. This research shows that TPC was significantly higher (P < 0.05) in green coffee compared to TPC in roasted coffee, and TPC decreases as the roasting temperature increases. TFC and FC were significantly lower (P < 0.05) in green coffee than in roasted coffee. Differences in TPC between the 1st and 2nd classes of Arabica coffee were not significant (P > 0.05), while differences in TFC were significant (P < 0.05) only for green coffee from the second production batch and differences in FC were significant (P < 0.05) for green coffee and for coffee roasted at 175°C. Roasting temperatures have different influences the antioxidant activity (DPPH, ABTS) of coffee and the highest antioxidant activity was determined in coffee roasted at 171°C. An exception was 1st class Arabica coffee roasted at 167°C (ABTS). All samples of 1st class Arabica coffee had higher antioxidant activity (DPPH, ABTS) compared to 2nd class Arabica. This research shows that the bioactive compounds content and antioxidant activity of different quality classes of Arabica coffee are dependent on the degree of roasting. TPC decreases when the roasting temperature increases, while TFC and FC also increase. These results indicate that the antioxidant activity of coffee depends on a variety of bioactive components in coffee beans. Antioxidant activity largely depends on the class of coffee. The coffee samples of 1stclass quality (maximum 8 black beans/300 g from the sample and large bean size) had higher antioxidant activity compared to samples of 2nd quality class (maximum 19 black beans/300 g in the sample and medium-sized beans).

Novel mesenchymal and haematopoietic cell isoforms of the SHP-2 docking receptor, PZR: identification, molecular cloning and effects on cell migration.

PubMed Central

Zannettino, Andrew C W; Roubelakis, Maria; Welldon, Katie J; Jackson, Denise E; Simmons, Paul J; Bendall, Linda J; Henniker, Anthony; Harrison, Kate L; Niutta, Silvana; Bradstock, Kenneth F; Watt, Suzanne M

2003-01-01

SHP-2 (Src homology phosphatase type-2) is essential for haematopoietic skeletal and vascular development. Thus the identification of its binding partners is critically important. In the present study, we describe a unique monoclonal antibody, WM78, which interacts with PZR, a SHP-2 binding partner. Furthermore, we identify two novel isoforms of PZR, PZRa and PZRb, derived by differential splicing from a single gene transcription unit on human chromosome 1q24. All are type 1 transmembrane glycoproteins with identical extracellular and transmembrane domains, but differ in their cytoplasmic tails. The PZR intracellular domain contains two SHP-2 binding immunoreceptor tyrosine-based inhibitory motifs (VIY(246)AQL and VVY(263)ADI) which are not present in PZRa and PZRb. Using the WM78 monoclonal antibody, which recognizes the common extracellular domain of the PZR isoforms, we demonstrate that the PZR molecules are expressed on mesenchymal and haematopoietic cells, being present on the majority of CD34(+)CD38(+) and early clonogenic progenitors, and at lower levels on CD34(+)CD38(-) cells and the hierarchically more primitive pre-colony forming units. Interestingly, we show by reverse transcriptase-PCR that the PZR isoforms are differentially expressed in haematopoietic, endothelial and mesenchymal cells. Both PZR and PZRb are present in CD133(+) precursors and endothelial cells, PZRb predominates in mesenchymal and committed myelomonocytic progenitor cells, and all three isoforms occur in erythroid precursor cell lines. Importantly, using SHP-2 mutant (Delta 46-110) and SHP-2 rescue of embryonic fibroblasts stably expressing the PZR isoforms, we demonstrate for the first time that PZR, but not PZRa or PZRb, facilitates fibronectin- dependent migration of cells expressing a competent SHP-2 molecule. These observations will be instrumental in determining the mechanisms whereby PZR isoforms regulate cell motility. PMID:12410637

Human PDE4D isoform composition is deregulated in primary prostate cancer and indicative for disease progression and development of distant metastases

PubMed Central

Böttcher, René; Dulla, Kalyan; van Strijp, Dianne; Dits, Natasja; Verhoef, Esther I.; Baillie, George S.; van Leenders, Geert J.L.H.; Houslay, Miles D.; Jenster, Guido; Hoffmann, Ralf

2016-01-01

Phosphodiesterase 4D7 was recently shown to be specifically over-expressed in localized prostate cancer, raising the question as to which regulatory mechanisms are involved and whether other isoforms of this gene family (PDE4D) are affected under the same conditions. We investigated PDE4D isoform composition in prostatic tissues using a total of seven independent expression datasets and also included data on DNA methylation, copy number and AR and ERG binding in PDE4D promoters to gain insight into their effect on PDE4D transcription. We show that expression of PDE4D isoforms is consistently altered in primary human prostate cancer compared to benign tissue, with PDE4D7 being up-regulated while PDE4D5 and PDE4D9 are down-regulated. Disease progression is marked by an overall down-regulation of long PDE4D isoforms, while short isoforms (PDE4D1/2) appear to be relatively unaffected. While these alterations seem to be independent of copy number alterations in the PDE4D locus and driven by AR and ERG binding, we also observed increased DNA methylation in the promoter region of PDE4D5, indicating a long lasting alteration of the isoform composition in prostate cancer tissues. We propose two independent metrics that may serve as diagnostic and prognostic markers for prostate disease: (PDE4D7 - PDE4D5) provides an effective means for distinguishing PCa from normal adjacent prostate, whereas PDE4D1/2 - (PDE4D5 + PDE4D7 + PDE4D9) offers strong prognostic potential to detect aggressive forms of PCa and is associated with metastasis free survival. Overall, our findings highlight the relevance of PDE4D as prostate cancer biomarker and potential drug target. PMID:27683107

A comparison of the enzymatic properties of three recombinant isoforms of thrombolytic and antibacterial protein--Destabilase-Lysozyme from medicinal leech.

PubMed

Kurdyumov, Alexey S; Manuvera, Valentin A; Baskova, Isolda P; Lazarev, Vassili N

2015-11-21

Destabilase-Lysozyme (mlDL) is a multifunctional i-type enzyme that has been found in the secretions from the salivary glands of medicinal leeches. mlDL has been shown to exhibit isopeptidase, muramidase and antibacterial activity. This enzyme attracts interest because it expresses thrombolytic activity through isopeptidolysis of the ε-(γ-Glu)-Lys bonds that cross-link polypeptide chains in stabilised fibrin. To date, three isoforms of mlDL have been identified. The enzymatic properties of pure mlDL isoforms have not yet been described because only destabilase complexes containing other proteins could be isolated from the salivary gland secretion and because low product yield from the generation of recombinant proteins has made comprehensive testing difficult. In the present study, we optimised the procedures related to the expression, isolation and purification of active mlDL isoforms (mlDL-Ds1, mlDL-Ds2, mlDL-Ds3) using an Escherichia coli expression system, and we detected and compared their muramidase, lytic, isopeptidase and antimicrobial activities. After optimisation, the product yield was 30 mg per litre of culture. The data obtained in our study led to the suggestion that the recombinant mlDL isoforms isolated from inclusion bodies form stable oligomeric complexes. Analyses of the tested activities revealed that all isoforms exhibited almost identical patterns of pH and ionic strength effects on the activities. We determined that mlDL-Ds1, 2, 3 possessed non-enzymatic antibacterial activity independent of their muramidase activity. For the first time, we demonstrated the fibrinolytic activity of the recombinant mlDL and showed that only intact proteins possessed this activity, suggesting their enzymatic nature. The recombinant Destabilase-Lysozyme isoforms obtained in our study may be considered potential thrombolytic agents that act through a mechanism different from that of common thrombolytics.

GSK3β isoform-selective regulation of depression, memory and hippocampal cell proliferation

PubMed Central

Pardo, Marta; Abrial, Erika; Jope, Richard S.; Beurel, Eleonore

2016-01-01

Abnormally active glycogen synthase kinase-3 (GSK3) contributes to pathological processes in multiple psychiatric and neurological disorders. Modeled in mice, this includes increasing susceptibility to dysregulation of mood-relevant behaviors, impairing performance in several cognitive tasks, and impairing adult hippocampal neural precursor cell (NPC) proliferation. These deficits are all evident in GSK3α/β knockin mice, in which serine-to-alanine mutations block the inhibitory serine phosphorylation regulation of both GSK3 isoforms, leaving GSK3 hyperactive. It was unknown if both GSK3 isoforms perform redundant actions in these processes, or if hyperactivity of one GSK3 isoform has a predominant effect. To test this, we examined GSK3α or GSK3β knockin mice in which only one isoform was mutated to a hyperactive form. Only GSK3β, not GSK3α, knockin mice displayed heightened vulnerability to the learned helplessness model of depression-like behavior. Three cognitive measures impaired in GSK3α/β knockin mice demonstrated differential regulation by GSK3 isoforms. Novel object recognition was impaired in GSK3β, not GSK3α, knockin mice, whereas temporal order memory was not impaired in GSK3α or GSK3β knockin mice, and coordinate spatial processing was impaired in both GSK3α and GSK3β knockin mice. Adult hippocampal NPC proliferation was severely impaired in GSK3β knockin mice, but not impaired in GSK3α knockin mice. Increased activity of GSK3β, in the absence of over-expression or disease pathology, is sufficient to impair mood regulation, novel object recognition, and hippocampal NPC proliferation, whereas hyperactive GSK3α individually does not impair these processes. These results demonstrate that hyperactivity of the two GSK3 isoforms execute non-redundant effects on these processes. PMID:26749572

GSK3β isoform-selective regulation of depression, memory and hippocampal cell proliferation.

PubMed

Pardo, M; Abrial, E; Jope, R S; Beurel, E

2016-03-01

Abnormally active glycogen synthase kinase-3 (GSK3) contributes to pathological processes in multiple psychiatric and neurological disorders. Modeled in mice, this includes increasing susceptibility to dysregulation of mood-relevant behaviors, impairing performance in several cognitive tasks and impairing adult hippocampal neural precursor cell (NPC) proliferation. These deficits are all evident in GSK3α/β knockin mice, in which serine-to-alanine mutations block the inhibitory serine phosphorylation regulation of both GSK3 isoforms, leaving GSK3 hyperactive. It was unknown if both GSK3 isoforms perform redundant actions in these processes, or if hyperactivity of one GSK3 isoform has a predominant effect. To test this, we examined GSK3α or GSK3β knockin mice in which only one isoform was mutated to a hyperactive form. Only GSK3β, not GSK3α, knockin mice displayed heightened vulnerability to the learned helplessness model of depression-like behavior. Three cognitive measures impaired in GSK3α/β knockin mice showed differential regulation by GSK3 isoforms. Novel object recognition was impaired in GSK3β, not in GSK3α, knockin mice, whereas temporal order memory was not impaired in GSK3α or GSK3β knockin mice, and co-ordinate spatial processing was impaired in both GSK3α and GSK3β knockin mice. Adult hippocampal NPC proliferation was severely impaired in GSK3β knockin mice, but not impaired in GSK3α knockin mice. Increased activity of GSK3β, in the absence of overexpression or disease pathology, is sufficient to impair mood regulation, novel object recognition and hippocampal NPC proliferation, whereas hyperactive GSK3α individually does not impair these processes. These results show that hyperactivity of the two GSK3 isoforms execute non-redundant effects on these processes. © 2016 John Wiley & Sons Ltd and International Behavioural and Neural Genetics Society.

Bidirectional regulation of neurite elaboration by alternatively spliced metabotropic glutamate receptor 5 (mGluR5) isoforms.

PubMed

Mion, S; Corti, C; Neki, A; Shigemoto, R; Corsi, M; Fumagalli, G; Ferraguti, F

2001-06-01

Alternative splicing in the mGluR5 gene generates two different receptor isoforms, of which expression is developmentally regulated. However, little is known about the functional significance of mGluR5 splice variants. We have examined the functional coupling, subcellular targeting, and effect on neuronal differentiation of epitope-tagged mGluR5 isoforms by expression in neuroblastoma NG108-15 cells. We found that both mGluR5 splice variants give rise to comparable [Ca2+]i transients and have similar pharmacological profile. Tagged receptors were shown by immunofluorescence to be inserted in the plasma membrane. In undifferentiated cells the subcellular localization of the two mGluR5 isoforms was partially segregated, whereas in differentiated cells the labeling largely redistributed to the newly formed neurites. Interestingly, we demonstrate that mGluR5 splice variants dramatically influence the formation and maturation of neurites; mGluR5a hinders the acquisition of mature neuronal traits and mGluR5b fosters the elaboration and extension of neurites. These effects are partly inhibited by MPEP. Copyright 2001 Academic Press.

The long and the short of SAD-1 kinase.

PubMed

Kim, Joanne S M; Hung, Wesley; Zhen, Mei

2010-05-01

The Ser/Thr SAD kinases are evolutionarily conserved, critical regulators of neural development. Exciting findings in recent years have significantly advanced our understanding of the mechanism through which SAD kinases regulate neural development. Mammalian SAD-A and SAD-B, activated by a master kinase LKB1, regulate microtubule dynamics and polarize neurons. In C. elegans, the sad-1 gene encodes two isoforms, namely the long and the short, which exhibit overlapping and yet distinct functions in neuronal polarity and synaptic organization. Surprisingly, our most recent findings in C. elegans revealed a SAD-1-independent LKB1 activity in neuronal polarity. We also found that the long SAD-1 isoform directly interacts with a STRADalpha pseudokinase, STRD-1, to regulate neuronal polarity and synaptic organization. We elaborate here a working model of SAD-1 in which the two isoforms dimer/oligomerize to form a functional complex, and STRD-1 clusters and localizes the SAD-1 complex to synapses. While the mechanistic difference between the vertebrate and invertebrate SAD kinases may be puzzling, a recent discovery of the functionally distinct SAD-B isoforms predicts that the difference likely arises from our incomplete understanding of the SAD kinase mechanism and may eventually be reconciled as the revelation continues.

Identification of a new B4GalNAcT1 (GM2/GD2/GA2 synthase) isoform, and regulation of enzyme stability and intracellular transport by arginine-based motif.

PubMed

Shishido, Fumi; Uemura, Satoshi; Kashimura, Madoka; Inokuchi, Jin-Ichi

2017-10-01

Glycosphingolipids (GSLs) are abundant in plasma membranes of mammalian cells, and their synthesis is strictly regulated in the Golgi apparatus. Disruption of GSL homeostasis is the cause of numerous diseases. Hundreds of molecular species of GSLs exist, and the detailed mechanisms underlying their homeostasis remain unclear. We investigated the physiological significance of isoform production for β1,4-N-acetyl-galactosaminyl transferase 1/B4GALNT1 (B4GN1), an enzyme involved in synthesis of ganglio-series GSLs GM2/GD2/GA2. We discovered a new mRNA variant (termed variant 2) of B4GN1 through EST clone search. A new isoform, M1-B4GN1, which has an NH 2 -terminal cytoplasmic tail longer than that of previously-known isoform M2-B4GN1, is translated from variant 2. M1-B4GN1 has R-based motif (a retrograde transport signal) in the cytoplasmic tail. M1-B4GN1 is partially localized in the endoplasmic reticulum (ER) depending on the R-based motif, whereas M2-B4GN1 is localized in the Golgi. Stability of M1-B4GN1 is higher than that of M2-B4GN1 because of the R-based motif. M2-B4GN1 forms a homodimer via disulfide bonding. When M1-B4GN1 and M2-B4GN1 were co-expressed in CHO-K1 cells, the two isoforms formed a heterodimer. The M1/M2-B4GN1 heterodimer was more stable than the M2-B4GN1 homodimer, but the heterodimer was not transported from the Golgi to the ER. Our findings indicate that stabilization of M1-B4GN1 homodimer and M1/M2-B4GN1 heterodimer by R-based motif is related to prolongation of Golgi retention, but not to retrograde transport from the Golgi to the ER. Coexistence of several B4GN1 isoforms having distinctive characteristics presumably helps maintain overall enzyme stability and GSL homeostasis. Copyright © 2017 Elsevier B.V. All rights reserved.

Alternative splicing of TIA-1 in human colon cancer regulates VEGF isoform expression, angiogenesis, tumour growth and bevacizumab resistance.

PubMed

Hamdollah Zadeh, Maryam A; Amin, Elianna M; Hoareau-Aveilla, Coralie; Domingo, Enric; Symonds, Kirsty E; Ye, Xi; Heesom, Katherine J; Salmon, Andrew; D'Silva, Olivia; Betteridge, Kai B; Williams, Ann C; Kerr, David J; Salmon, Andrew H J; Oltean, Sebastian; Midgley, Rachel S; Ladomery, Michael R; Harper, Steven J; Varey, Alexander H R; Bates, David O

2015-01-01

The angiogenic capability of colorectal carcinomas (CRC), and their susceptibility to anti-angiogenic therapy, is determined by expression of vascular endothelial growth factor (VEGF) isoforms. The intracellular protein T-cell Intracellular Antigen (TIA-1) alters post-transcriptional RNA processing and binds VEGF-A mRNA. We therefore tested the hypothesis that TIA-1 could regulate VEGF-A isoform expression in colorectal cancers. TIA-1 and VEGF-A isoform expression was measured in colorectal cancers and cell lines. We discovered that an endogenous splice variant of TIA-1 encoding a truncated protein, short TIA-1 (sTIA-1) was expressed in CRC tissues and invasive K-Ras mutant colon cancer cells and tissues but not in adenoma cell lines. sTIA-1 was more highly expressed in CRC than in normal tissues and increased with tumour stage. Knockdown of sTIA-1 or over-expression of full length TIA-1 (flTIA-1) induced expression of the anti-angiogenic VEGF isoform VEGF-A165b. Whereas flTIA-1 selectively bound VEGF-A165 mRNA and increased translation of VEGF-A165b, sTIA-1 prevented this binding. In nude mice, xenografted colon cancer cells over-expressing flTIA-1 formed smaller, less vascular tumours than those expressing sTIA-1, but flTIA-1 expression inhibited the effect of anti-VEGF antibodies. These results indicate that alternative splicing of an RNA binding protein can regulate isoform specific expression of VEGF providing an added layer of complexity to the angiogenic profile of colorectal cancer and their resistance to anti-angiogenic therapy. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

Recent developments in track reconstruction and hadron identification at MPD

NASA Astrophysics Data System (ADS)

Mudrokh, A.; Zinchenko, A.

2017-03-01

A Monte Carlo simulation of real detector effects with as many details as possible has been carried out instead of a simplified Geant point smearing approach during the study of the detector performance. Some results of realistic simulation of the MPD TPC (Time Projection Chamber) including digitization in central Au+Au collisions have been obtained. Particle identification (PID) has been tuned to account for modifications in the track reconstruction. Some results on hadron identification in the TPC and TOF (Time Of Flight) detectors with realistically simulated response have been also obtained.

Danon disease – dysregulation of autophagy in a multisystem disorder with cardiomyopathy

PubMed Central

Rowland, Teisha J.; Sweet, Mary E.; Mestroni, Luisa; Taylor, Matthew R. G.

2016-01-01

ABSTRACT Danon disease is a rare, severe X-linked form of cardiomyopathy caused by deficiency of lysosome-associated membrane protein 2 (LAMP-2). Other clinical manifestations include skeletal myopathy, cognitive defects and visual problems. Although individuals with Danon disease have been clinically described since the early 1980s, the underlying molecular mechanisms involved in pathological progression remain poorly understood. LAMP-2 is known to be involved in autophagy, and a characteristic accumulation of autophagic vacuoles in the affected tissues further supports the idea that autophagy is disrupted in this disease. The LAMP2 gene is alternatively spliced to form three splice isoforms, which are thought to play different autophagy-related cellular roles. This Commentary explores findings from genetic, histological, functional and tissue expression studies that suggest that the specific loss of the LAMP-2B isoform, which is likely to be involved in macroautophagy, plays a crucial role in causing the Danon phenotype. We also compare findings from mouse and cellular models, which have allowed for further molecular characterization but have also shown phenotypic differences that warrant attention. Overall, there is a need to better functionally characterize the LAMP-2B isoform in order to rationally explore more effective therapeutic options for individuals with Danon disease. PMID:27165304

Cyt c6-3: A New Isoform of Photosynthetic Cyt c6 Exclusive to Heterocyst-Forming Cyanobacteria.

PubMed

Torrado, Alejandro; Valladares, Ana; Puerto-Galán, Leonor; Hervás, Manuel; Navarro, José A; Molina-Heredia, Fernando P

2017-02-01

All known cyanobacteria contain Cyt c6, a small soluble electron carrier protein whose main function is to transfer electrons from the Cyt b6f complex to PSI, although it is also involved in respiration. We have previously described a second isoform of this protein, the Cyt c6-like, whose function remains unknown. Here we describe a third isoform of Cyt c6 (here called Cytc6-3), which is only found in heterocyst-forming filamentous cyanobacteria. Cyt c6-3 is expressed in vegetative cells but is specifically repressed in heterocysts cells under diazotrophic growth conditions. Although there is a close structural similarity between Cyt c6-3 and Cyt c6 related to the general protein folding, Cyt c6-3 presents differential electrostatic surface features as compared with Cyt c6, its expression is not copper dependent and has a low reactivity towards PSI. According to the different expression pattern, functional reactivity and structural properties, Cyt c6-3 has to play an as yet to be defined regulatory role related to heterocyst differentiation. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

[Neuromuscular system and aging: involutions and implications].

PubMed

Paillard, Thierry

2013-12-01

In aged human, the number of muscle fibers and motor units decreases. The remaining motor units lose their functionality (decrease of the discharge frequency, greater fluctuation of the discharge) particularly those which contain type II fibers. The renewal of intracellular proteins declines which creates a negative balance between the daily protein losses and the capacities to renew them. The activity of the protein kinase (Akt) that stimulates the synthesis of regulation proteins (mTOR, p70S6, IGFBP-5) declines whereas the factors of degradation of proteins (NF-kappa B) are activated. Besides, the process of activation and proliferation of satellite cells is affected and the production of anabolic hormones and local factors is decreased. After a strength training program, muscle hypertrophy is linked to the protein synthesis at the level of myosin heavy chain (MHC) isoforms in older subjects. However, the transcription of the genes that code the MHC-I (slow form) increases and the transcription of the genes that code the MHC-II (fast form) decreases. Thus, the transition of the phenotype towards a slower form cannot be inverted by strength training during the advanced in age. Moreover, strength training enables to decrease the proportion of fibers containing MHC of hybrid form in the process of evolution. Hence, strength training can engender a stabilization of the muscular phenotype i.e. different isoforms of MHC. In addition, strength training counteracts the noxious effects mentioned above by generating muscular hypertrophy thanks to a reactive increase in the production of anabolic hormones. A program of aerobic training can induce an increase in the synthesis of ARN messengers coding isoforms related to the oxidative metabolism (MHC-I and to a lesser extent MHC-IIa) while the transcribed for the type MHC-IIx decrease.

Alternative Polyadenylation in Glioblastoma Multiforme and Changes in Predicted RNA Binding Protein Profiles

PubMed Central

Shao, Jiaofang; Zhang, Jing; Zhang, Zengming; Jiang, Huawei; Lou, Xiaoyan; Foltz, Gregory; Lan, Qing; Huang, Qiang

2013-01-01

Abstract Alternative polyadenylation (APA) is widely present in the human genome and plays a key role in carcinogenesis. We conducted a comprehensive analysis of the APA products in glioblastoma multiforme (GBM, one of the most lethal brain tumors) and normal brain tissues and further developed a computational pipeline, RNAelements (http://sysbio.zju.edu.cn/RNAelements/), using covariance model from known RNA binding protein (RBP) targets acquired by RNA Immunoprecipitation (RIP) analysis. We identified 4530 APA isoforms for 2733 genes in GBM, and found that 182 APA isoforms from 148 genes showed significant differential expression between normal and GBM brain tissues. We then focused on three genes with long and short APA isoforms that show inconsistent expression changes between normal and GBM brain tissues. These were myocyte enhancer factor 2D, heat shock factor binding protein 1, and polyhomeotic homolog 1 (Drosophila). Using the RNAelements program, we found that RBP binding sites were enriched in the alternative regions between the first and the last polyadenylation sites, which would result in the short APA forms escaping regulation from those RNA binding proteins. To the best of our knowledge, this report is the first comprehensive APA isoform dataset for GBM and normal brain tissues. Additionally, we demonstrated a putative novel APA-mediated mechanism for controlling RNA stability and translation for APA isoforms. These observations collectively lay a foundation for novel diagnostics and molecular mechanisms that can inform future therapeutic interventions for GBM. PMID:23421905

RAGE splicing variants in mammals.

PubMed

Sterenczak, Katharina Anna; Nolte, Ingo; Murua Escobar, Hugo

2013-01-01

The receptor for advanced glycation end products (RAGE) is a multiligand receptor of environmental stressors which plays key roles in pathophysiological processes, including immune/inflammatory disorders, Alzheimer's disease, diabetic arteriosclerosis, tumorigenesis, and metastasis. Besides the full-length RAGE protein in humans nearly 20 natural occurring RAGE splicing variants were described on mRNA and protein level. These naturally occurring isoforms are characterized by either N-terminally or C-terminally truncations and are discussed as possible regulators of the full-length RAGE receptor either by competitive ligand binding or by displacing the full-length protein in the membrane. Accordingly, expression deregulations of the naturally occurring isoforms were supposed to have significant effect on RAGE-mediated disorders. Thereby the soluble C-truncated RAGE isoforms present in plasma and tissues are the mostly focused isoforms in research and clinics. Deregulations of the circulating levels of soluble RAGE forms were reported in several RAGE-associated pathological disorders including for example atherosclerosis, diabetes, renal failure, Alzheimer's disease, and several cancer types. Regarding other mammalian species, the canine RAGE gene showed high similarities to the corresponding human structures indicating RAGE to be evolutionary highly conserved between both species. Similar to humans the canine RAGE showed a complex and extensive splicing activity leading to a manifold pattern of RAGE isoforms. Due to the similarities seen in several canine and human diseases-including cancer-comparative structural and functional analyses allow the development of RAGE and ligand-specific therapeutic approaches beneficial for human and veterinary medicine.

A computational analysis of the three isoforms of glutamate dehydrogenase reveals structural features of the isoform EC 1.4.1.4 supporting a key role in ammonium assimilation by plants

PubMed Central

Jaspard, Emmanuel

2006-01-01

Background There are three isoforms of glutamate dehydrogenase. The isoform EC 1.4.1.4 (GDH4) catalyses glutamate synthesis from 2-oxoglutarate and ammonium, using NAD(P)H. Ammonium assimilation is critical for plant growth. Although GDH4 from animals and prokaryotes are well characterized, there are few data concerning plant GDH4, even from those whose genomes are well annotated. Results A large set of the three GDH isoforms was built resulting in 116 non-redundant full polypeptide sequences. A computational analysis was made to gain more information concerning the structure – function relationship of GDH4 from plants (Eukaryota, Viridiplantae). The tested plant GDH4 sequences were the two ones known to date, those of Chlorella sorokiniana. This analysis revealed several structural features specific of plant GDH4: (i) the lack of a structure called "antenna"; (ii) the NAD(P)-binding motif GAGNVA; and (iii) a second putative coenzyme-binding motif GVLTGKG together with four residues involved in the binding of the reduced form of NADP. Conclusion A number of structural features specific of plant GDH4 have been found. The results reinforce the probable key role of GDH4 in ammonium assimilation by plants. Reviewers This article was reviewed by Tina Bakolitsa (nominated by Eugene Koonin), Martin Jambon (nominated by Laura Landweber), Sandor Pangor and Franck Eisenhaber. PMID:17173671

Synthesis and Biological Evaluation of 4-Sulfamoylphenyl/Sulfocoumarin Carboxamides as Selective Inhibitors of Carbonic Anhydrase Isoforms hCA II, IX, and XII.

PubMed

Angapelly, Srinivas; Angeli, Andrea; Khan, Arbaj Jabbar; Sri Ramya, P V; Supuran, Claudiu T; Arifuddin, Mohammed

2018-04-19

With the aim to develop potent and selective human carbonic anhydrase inhibitors (hCAIs), we synthesized 4-sulfamoylphenyl/sulfocoumarin benzamides (series 5 a-r and series 7 a-q) and evaluated their inhibition profiles against five isoforms of the zinc-containing human carbonic anhydrase (hCA, EC 4.2.1.1): cytosolic hCA I and II, and the transmembrane isozymes hCA IV, IX, and XII. Compounds 5 a-r were found to selectively inhibit hCA II in the nanomolar range, while being less effective against the other hCA isoforms. As noted from the literature, sulfocoumarin (1,2-benzoxathiine 2,2-dioxide) acts as a "prodrug" inhibitor and is hydrolyzed by the esterase activity of hCA to form 2-hydroxyphenylvinylsulfonic acid, which thereafter binds to the enzyme in a manner similar to that of coumarins and sulfoxocoumarins. All these sulfocoumarins (compounds 7 a-q) were found to be very weak or ineffective as inhibitors of the housekeeping off-target hCA isoforms I and II, and effectively inhibited the transmembrane tumor-associated isoforms IX and XII in the high nanomolar to micromolar ranges. Further structural modifications of these molecules could be useful for the development of effective hCA inhibitors used for the treatment of glaucoma, epilepsy, and cancer. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Effect of Roasting Process on Total Phenolic Compounds and γ-tocopherol Contents of Iranian Sesame Seeds (Sesamum indicum)

PubMed Central

Jannat, Behrooz; Oveisi, Mohammad Reza; Sadeghi, Naficeh; Hajimahmoodi, Mannan; Behzad, Masoomeh; Nahavandi, Bahman; Tehrani, Shirin; Sadeghi, Fatemeh; Oveisi, Morvarid

2013-01-01

Sesame (Sesamum indicum L.) seed and oil have long been used widely as healthy foods to supply energy and prevent aging. Some of the main active anti-oxidative constituents in sesame seeds are γ-tocopherol and phenols. The purpose of this study was to investigate the relationship between roasting temperature and time with γ-tocopherol and total phenolic compounds (TPC) of sesame seeds when roasted in a domestic electric oven. Eight cultivars of sesame seeds in this study were Darab, Dezful, Karaj, Moghan, Naz- Branching, Naz-NonBranching, Siah and Varamin. Each cultivar was divided into ten group based on the roasting time (10, 15 and 20 min) and temperatures (180, 200 and 220 °C)andunroasted one. The high-performance liquid chromatography (HPLC) and spectrophotometeric methods were used for γ-tocopherol (n = 80) and TPC (n = 80) analysis, respectively. The γ-tocopherol content ranged from 329 ± 5 mg/L in Naz-Branching sesame oil to 1114±7 mg/L in Siah sesame oil and 169±6 to 577±1 mg/kg in sesame seed respectively. γ-tocopherol content of six cultivars increased significantly (p < 0.05) as the roasting temperature and time; until 200 °C for 10 min, but they were decreased by roasting at 220 °C in longer time. Also TPC increased significantly as the roasting temperature. The amount of TPC varied in different sesame cultivars from 20.109 ± 3.967 μM to 129.300±3.493 in Varamin and Naz- Branching sesame seed cultivars, respectively, also TPC increased from 70.953 ± 5.863 μM in unroasted Naz-Branching sesame seed to 129.300 ± 3.493 μM after roasting in 200 °C for 20 min. The present study showed that Iranian sesame seed can be considered as a good source of natural antioxidant specially after roasting. The optimum temperature and time roasting to obtain the most γ-tocopherol and total phenolic content was 200 °C for 10 and 20 min, respectively. PMID:24523755

Effect of Roasting Process on Total Phenolic Compounds and γ-tocopherol Contents of Iranian Sesame Seeds (Sesamum indicum).

PubMed

Jannat, Behrooz; Oveisi, Mohammad Reza; Sadeghi, Naficeh; Hajimahmoodi, Mannan; Behzad, Masoomeh; Nahavandi, Bahman; Tehrani, Shirin; Sadeghi, Fatemeh; Oveisi, Morvarid

2013-01-01

Sesame (Sesamum indicum L.) seed and oil have long been used widely as healthy foods to supply energy and prevent aging. Some of the main active anti-oxidative constituents in sesame seeds are γ-tocopherol and phenols. The purpose of this study was to investigate the relationship between roasting temperature and time with γ-tocopherol and total phenolic compounds (TPC) of sesame seeds when roasted in a domestic electric oven. Eight cultivars of sesame seeds in this study were Darab, Dezful, Karaj, Moghan, Naz- Branching, Naz-NonBranching, Siah and Varamin. Each cultivar was divided into ten group based on the roasting time (10, 15 and 20 min) and temperatures (180, 200 and 220 °C)andunroasted one. The high-performance liquid chromatography (HPLC) and spectrophotometeric methods were used for γ-tocopherol (n = 80) and TPC (n = 80) analysis, respectively. The γ-tocopherol content ranged from 329 ± 5 mg/L in Naz-Branching sesame oil to 1114±7 mg/L in Siah sesame oil and 169±6 to 577±1 mg/kg in sesame seed respectively. γ-tocopherol content of six cultivars increased significantly (p < 0.05) as the roasting temperature and time; until 200 °C for 10 min, but they were decreased by roasting at 220 °C in longer time. Also TPC increased significantly as the roasting temperature. The amount of TPC varied in different sesame cultivars from 20.109 ± 3.967 μM to 129.300±3.493 in Varamin and Naz- Branching sesame seed cultivars, respectively, also TPC increased from 70.953 ± 5.863 μM in unroasted Naz-Branching sesame seed to 129.300 ± 3.493 μM after roasting in 200 °C for 20 min. The present study showed that Iranian sesame seed can be considered as a good source of natural antioxidant specially after roasting. The optimum temperature and time roasting to obtain the most γ-tocopherol and total phenolic content was 200 °C for 10 and 20 min, respectively.

Status of the R&D activities for the upgrade of the ALICE TPC

NASA Astrophysics Data System (ADS)

Deisting, Alexander

2018-02-01

After the Long Shutdown 2 (LS2) the LHC will provide lead-lead collisions at interaction rates as high as 50 kHz. In order to cope with such conditions the ALICE Time Projection Chamber (TPC) needs to be upgraded. After the upgrade the TPC will run in a continuous mode, without any degradation of the momentum and dE/dx resolution compared to the performance of the present TPC. Since readout by multi-wire proportional chambers is no longer feasible with these requirements, new technologies have to be employed. In the new readout chambers the electron amplification is provided by a stack of four Gas ElectronMultiplier (GEM) foils. Here foils with a standard hole pitch of 140 μm as well as large pitch foils (280 μm) are used. Their high voltage settings and orientation have been optimised to provide an energy resolution of σE/E ≤ 12% at the photopeak of 55Fe. At the same settings the Ion BackFlow into the drift volume is less than 1% of the effective number of ions produced during gas amplification and the primary ionisations. This is necessary to prevent the accumulation of space charge, which eventually will distort the field in the drift volume. To ensure stable operation at the high loads during LHC run 3 the chambers have to be robust against discharges, too. With the selected configuration in a quadruple GEMstack the discharge probability is kept at the level of 10-12 discharges per incoming hadron. An overview of the ALICE TPC upgrade activities will be given in these proceedings and the optimised settings foreseen for the GEM stacks of the future readout chambers are introduced. Furthermore the outcome of two beam time campaigns at SPS and PS (at CERN) in the end of 2014 is shown. At this campaigns the stability against discharges and the dE/dx performance of a full size readout chamber prototype was tested. In addition it is reported on charging-up studies of 4GEM stacks and on tests of electromagnetic sagging of large GEM foils.

Molecular analysis of alternative transcripts of equine AXL receptor tyrosine kinase gene.

PubMed

Park, Jeong-Woong; Song, Ki-Duk; Kim, Nam Young; Choi, Jae-Young; Hong, Seul A; Oh, Jin Hyeog; Kim, Si Won; Lee, Jeong Hyo; Park, Tae Sub; Kim, Jin-Kyoo; Kim, Jong Geun; Cho, Byung-Wook

2017-10-01

Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified AXL receptor tyrosine kinase ( AXL ) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. We validated two isoforms of AXL transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Both of AXLa and AXLb transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between AXLa and AXLb transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. It is assumed that the expression patterns of AXLa and AXLb transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an NF-κB signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.

FPGA based data processing in the ALICE High Level Trigger in LHC Run 2

NASA Astrophysics Data System (ADS)

Engel, Heiko; Alt, Torsten; Kebschull, Udo; ALICE Collaboration

2017-10-01

The ALICE High Level Trigger (HLT) is a computing cluster dedicated to the online compression, reconstruction and calibration of experimental data. The HLT receives detector data via serial optical links into FPGA based readout boards that process the data on a per-link level already inside the FPGA and provide it to the host machines connected with a data transport framework. FPGA based data pre-processing is enabled for the biggest detector of ALICE, the Time Projection Chamber (TPC), with a hardware cluster finding algorithm. This algorithm was ported to the Common Read-Out Receiver Card (C-RORC) as used in the HLT for RUN 2. It was improved to handle double the input bandwidth and adjusted to the upgraded TPC Readout Control Unit (RCU2). A flexible firmware implementation in the HLT handles both the old and the new TPC data format and link rates transparently. Extended protocol and data error detection, error handling and the enhanced RCU2 data ordering scheme provide an improved physics performance of the cluster finder. The performance of the cluster finder was verified against large sets of reference data both in terms of throughput and algorithmic correctness. Comparisons with a software reference implementation confirm significant savings on CPU processing power using the hardware implementation. The C-RORC hardware with the cluster finder for RCU1 data is in use in the HLT since the start of RUN 2. The extended hardware cluster finder implementation for the RCU2 with doubled throughput is active since the upgrade of the TPC readout electronics in early 2016.

Sequential high pressure extractions applied to recover piceatannol and scirpusin B from passion fruit bagasse.

PubMed

Viganó, Juliane; Aguiar, Ana C; Moraes, Damila R; Jara, José L P; Eberlin, Marcos N; Cazarin, Cinthia B B; Maróstica, Mário R; Martínez, Julian

2016-07-01

Passion fruit seeds are currently discarded on the pulp processing but are known for their high piceatannol and scirpusin B contents. Using pressurized liquid extraction (PLE), these highly valuable phenolic compounds were efficiently extracted from defatted passion fruit bagasse (DPFB). PLE was performed using mixtures of ethanol and water (50 to 100% ethanol, w/w) as solvent, temperatures from 50 to 70°C and pressure at 10MPa. The extraction methods were compared in terms of the global yield, total phenolic content (TPC), piceatannol content and the antioxidant capacity of the extracts. The DPFB extracts were also compared with those from non-defatted passion fruit bagasse (nDPFB). Identification and quantification of piceatannol were performed using UHPLC-MS/MS. The results showed that high TPC and piceatannol content were achieved for the extracts obtained from DPFB through PLE at 70°C and using 50 and 75% ethanol as the solvent. The best PLE conditions for TPC (70°C, 75% ethanol) resulted in 55.237mgGAE/g dried and defatted bagasse, whereas PLE at 70°C and 50% ethanol achieved 18.590mg of piceatannol/g dried and defatted bagasse, and such yields were significantly higher than those obtained using conventional extraction techniques. The antioxidant capacity assays showed high correlation with the TPC (r>0.886) and piceatannol (r>0.772). The passion fruit bagasse has therefore proved to be a rich source of piceatannol and PLE showed high efficiency to recover phenolic compounds from defatted passion fruit bagasse. Copyright © 2016 Elsevier Ltd. All rights reserved.

Prevalence of fibromyalgia in Turkish geriatric population and its impact on quality of life.

PubMed

Garip, Yeşim; Öztaş, Dilek; Güler, Tuba

2016-10-01

The aim of the present study was to examine the presence of fibromyalgia (FM) in elderly adults and to evaluate the impact of the severity of FM on quality of life. A total of 100 patients between 65 and 80 years of age were included. The main admission diagnosis of the patients was recorded. Presence of FM was evaluated based on 1990 American College of Rheumatology (ACR) diagnostic criteria. The FM group was comprised of 31 patients fulfilling these criteria, and the remaining 69 patients composed the non-FM group. Tender point count (TPC) and common symptoms were recorded. FM disease severity was assessed using Fibromyalgia Impact Questionnaire (FIQ). Nottingham Health Profile (NHP) was used to evaluate quality of life. Pain severity was measured using Visual Analog Scale (VAS). Rate of FM was found to be 31%. FM patients scored significantly higher on pain, sleep, social isolation, and emotional reactions subgroups of NHP when compared to controls (p<0.05). TPC and FIQ were not affected by gender difference (p>0.05), but reduced with increasing age (p<0.01). FIQ and TPC were found to be correlated with only the pain and emotional reactions subgroups of NHP (p<0.01). There was no statistically significant correlation between FIQ and TPC and the physical mobility, sleep, energy, and social isolation subgroups of NHP (p>0.05). Although FM is known as a disease of young and middle-aged women, our study indicates that its prevalence increases with age. FM is associated with poor quality of life in terms of pain, sleep, social, and emotional functions.

Total phenolics content, anthocyanins, and dietary fiber content of apple pomace powders produced by vacuum-belt drying.

PubMed

Yan, Huitong; Kerr, William L

2013-04-01

Apple pomace is a waste material from apple juice processing, and contains significant amounts of dietary fiber and phytochemicals. Many of these compounds may be degraded post-pressing and during drying operations. Continuous vacuum-belt drying (VBD) was studied as a means of drying and maintaining quality of apple pomace. The color and chemical properties of samples dried by vacuum-belt drying at different temperatures were evaluated including total phenolics content (TPC), monomeric anthocyanins (TMA) and dietary fiber content (TDF). VBD powders were pale golden yellow, and those dried at 80°C did not differ in L*, a* and b* values from freeze-dried powders. VBD pomace had 44.9 to 51.9 g gallic acid equivalents kg(-1) TPC, with greatest retention for pomace dried at 80 and 95°C. TPC for pomace dried at 80 or 95°C was not significantly different from that for freeze-dried pomace. TMA levels (74.0 mg C3G kg(-1), where C3G is cyanidine 3-O-glucoside equivalents) were highest in pomace vacuum dried at 80°C. TDF ranged from 442 to 495 g kg(-1) in vacuum-dried pomace and was not significantly different from TDF of freeze-dried poamce (480 g kg(-1)). In all cases, TPC, TMA and TDF were higher in VBD pomace than in freeze-dried whole apple, while VBD pomace prepared at 80 or 95°C had fiber and phytochemical levels similar to freeze-dried powders. © 2012 Society of Chemical Industry.

Total Probability of Collision as a Metric for Finite Conjunction Assessment and Collision Risk Management

NASA Astrophysics Data System (ADS)

Frigm, R.; Johnson, L.

The Probability of Collision (Pc) has become a universal metric and statement of on-orbit collision risk. Although several flavors of the computation exist and are well-documented in the literature, the basic calculation requires the same input: estimates for the position, position uncertainty, and sizes of the two objects involved. The Pc is used operationally to make decisions on whether a given conjunction poses significant collision risk to the primary object (or space asset of concern). It is also used to determine necessity and degree of mitigative action (typically in the form of an orbital maneuver) to be performed. The predicted post-maneuver Pc also informs the maneuver planning process into regarding the timing, direction, and magnitude of the maneuver needed to mitigate the collision risk. Although the data sources, techniques, decision calculus, and workflows vary for different agencies and organizations, they all have a common thread. The standard conjunction assessment and collision risk concept of operations (CONOPS) predicts conjunctions, assesses the collision risk (typically, via the Pc), and plans and executes avoidance activities for conjunctions as a discrete events. As the space debris environment continues to increase and improvements are made to remote sensing capabilities and sensitivities to detect, track, and predict smaller debris objects, the number of conjunctions will in turn continue to increase. The expected order-of-magnitude increase in the number of predicted conjunctions will challenge the paradigm of treating each conjunction as a discrete event. The challenge will not be limited to workload issues, such as manpower and computing performance, but also the ability for satellite owner/operators to successfully execute their mission while also managing on-orbit collision risk. Executing a propulsive maneuver occasionally can easily be absorbed into the mission planning and operations tempo; whereas, continuously planning evasive maneuvers for multiple conjunction events is time-consuming and would disrupt mission and science operations beyond what is tolerable. At the point when the number of conjunctions is so large that it is no longer possible to consider each individually, some sort of an amalgamation of events and risk must be considered. This shift is to one where each conjunction cannot be treated individually and the effects of all conjunctions within a given period of time must be considered together. This new paradigm is called finite Conjunction Assessment (CA) risk management. This paper considers the use of the Total Probability of Collision (TPc) as an analogous collision risk metric in the finite CA paradigm. TPc is expressed by the equation below and provides an aggregate probability of colliding with any one of the predicted conjunctions under consideration. TPc=1-?(1-Pc,i) While the TPc computation is straightforward and its physical meaning is understandable, the implications of its usage operationally requires a change in mindset and approach to collision risk management. This paper explores the necessary changes to evolve the basic CA and collision risk management CONOPS from discrete to finite CA, including aspects of collision risk assessment and collision risk mitigation. It proposes numerical and graphical decision aids to understand both the “risk outlook” for a given primary as well as mitigation options for the total collision risk. Both concepts make use of the TPc as a metric for finite collision risk management. Several operational scenarios are used to demonstrate the proposed concepts in practice.

A change in fault-plane orientation between foreshocks and aftershocks of the Galway Lake earthquake, ML = 5.2, 1975, Mojave desert, California

USGS Publications Warehouse

Fuis, G.S.; Lindh, A.G.

1979-01-01

A marked change is observed in P/SV amplitude ratios, measured at station TPC, from foreshocks to aftershocks of the Galway Lake earthquake. This change is interpreted to be the result of a change in fault-plane orientation occurring between foreshocks and aftershocks. The Galway Lake earthquake, ML= 5.2, occurred on June 1, 1975. The first-motion fault-plane solutions for the main shock and most foreshocks and aftershocks indicate chiefly right-lateral strike-slip on NNW-striking planes that dip steeply, 70-90??, to the WSW. The main event was preceded by nine located foreshocks, ranging in magnitude from 1.9 to 3.4, over a period of 12 weeks, starting on March 9, 1975. All of the foreshocks form a tight cluster approximately 1 km in diameter. This cluster includes the main shock. Aftershocks are distributed over a 6-km-long fault zone, but only those that occurred inside the foreshock cluster are used in this study. Seismograms recorded at TPC (?? = 61 km), PEC (?? = 93 km), and CSP (?? = 83 km) are the data used here. The seismograms recorded at TPC show very consistent P/SV amplitude ratios for foreshocks. For aftershocks the P/SV ratios are scattered, but generally quite different from foreshock ratios. Most of the scatter for the aftershocks is confined to the two days following the main shock. Thereafter, however, the P/SV ratios are consistently half as large as for foreshocks. More subtle (and questionable) changes in the P/SV ratios are observed at PEC and CSP. Using theoretical P/SV amplitude ratios, one can reproduce the observations at TPC, PEC and CSP by invoking a 5-12?? counterclockwise change in fault strike between foreshocks and aftershocks. This interpretation is not unique, but it fits the data better than invoking, for example, changes in dip or slip angle. First-motion data cannot resolve this small change, but they permit it. Attenuation changes would appear to be ruled out by the fact that changes in the amplitude ratios, PTPC/PPEC and ptpc/pcsp, are observed, and these changes accompany the changes in P/SV. Observations for the Galway Lake earthquake are similar to observations for the Oroville, California, earthquake (ML = 5.7) of August 1, 1975, and the Brianes Hills, California, earthquake (ML = 4.3) of January 8, 1977 (Lindh et al., Science Vol. 201, pp. 56-59). A change in fault-plane orientation between foreshocks and aftershocks may be understandable in terms of early en-echelon cracking (foreshocks) giving way to shear on the main fault plane (main shock plus aftershocks). Recent laboratory data (Byerlee et al., Tectonophysics, Vol. 44, pp. 161-171) tend to support this view. ?? 1979.

Validation of an Italian version of the Fibromyalgia Impact Questionnaire (FIQ-I).

PubMed

Sarzi-Puttini, P; Atzeni, F; Fiorini, T; Panni, B; Randisi, G; Turiel, M; Carrabba, M

2003-01-01

To validate a translated Italian version of the Fibromyalgia Impact Questionnaire (FIQ). The Italian version of the FIQ was administered to 50 patients affected by fibromyalgia (FM) (48 patients filled out the questionnaire again 10 days later) together with the Italian version of the Stanford Health Assessment Questionnaire (HAQ), the Medical Outcomes Survey Short Form-36 (SF-36), and a tender point count (TPC) obtained by summing the score (0-3) of each tender point tested by thumb palpation. All patients were asked about the severity of pain today (10 cm visual analog scale) and the duration of symptoms. Test-retest reliability was assessed using Spearman correlations. Internal consistency was evaluated with Cronbach's alpha of reliability. Construct validity of the FIQ was evaluated by correlations between the HAQ and subscales of the SF-36 as well as the TPC. The mean duration of symptoms was 6.5 years and the mean age of the participants was 57.4 years. Test-retest reliability was between 0.74 and 0.95 for physical functioning as well as for the total FIQ and other components. Internal consistency was 0.90 for the overall FIQ. Significant correlations were obtained between the FIQ items, the HAQ and the SF-36. The Italian FIQ is a reliable and valid instrument for detecting and measuring functional disability and health status in Italian patients with FM.

Phytosynthesis of Iron Nanoparticle from Averrhoa Bilimbi Linn.

NASA Astrophysics Data System (ADS)

Rosli, I. R.; Zulhaimi, H. I.; Ibrahim, S. K. M.; Gopinath, S. C. B.; Kasim, K. F.; Akmal, H. M.; Nuradibah, M. A.; Sam, T. S.

2018-03-01

This paper demonstrates iron nanoparticles (FeNP) was synthesized from natural sources of Averrhoa bilimbi Linn. The plant extracts act as natural reducing agent in producing FeNP. There is no addition of any surfactants during the nanoparticles formation. Gravimetric analysis is used to calculate the percentage yield of plant extracts. TPC and DPPH assay method were used to evaluate antioxidant activity in different A. bilimbi extracts and synthesized FeNP. Based on the analyses, it showed that fruit has the highest percentage yield and antioxidant activity followed by leaf, twig and bark. Analysis from TPC, fruit contains 27.26 mg GAE/g and 39.46 mg GAE/g for FeNP. DPPH assay showed fruit extract has the highest free radical antioxidant activity with 61.93% in A. bilimbi and 80.00% in FeNP. Phytosynthesis of FeNP were examine by using UV-Vis spectrophotometer. Based on the spectra, it showed that FeNP recorded peak absorbance at 465 nm, 450 nm, 460 nm and 440 nm for UAE-F, UAE-L, UAE-T and UAE-B, respectively. FTIR analysis shows the presence of strong alcoholic bond, aldehyde, stretch amine and alkene that was responsible in reduction process to form FeNP. The result of UV-Vis and FTIR showed that the existance of FeNP and involvement of functional group that were responsible on the formation of nanoparticles.

Measuring Fission Fragment Mass Distributions as a Function of Incident Neutron Energy Using the fissionTPC

NASA Astrophysics Data System (ADS)

Gearhart, Joshua; Niffte Collaboration

2017-09-01

Fission fragment mass distributions are important observables for developing next generation dynamical models of fission. Many previous measurements have utilized ionization chambers to measure fission fragment energies and emission angles which are then used for mass calculations. The Neutron Induced Fission Fragment Tracking Experiment (NIFFTE) collaboration has built a time projection chamber (fissionTPC) that is capable of measuring additional quantities such as the ionization profiles of detected particles, allowing for the association of an individual fragment's ionization profile with its mass. The fragment masses are measured using the previously established 2E method. The fissionTPC takes its data using a continuous incident neutron energy spectrum provided by the Los Alamos Neutron Science CEnter (LANSCE). Mass distribution measurements across a continuous range of neutron energies put stronger constraints on fission models than similar measurements conducted at a handful of discrete neutron energies. This material is based upon work supported by the Department of Energy National Nuclear Security Administration under Award Numbers DE-NA0003180 and DE-NA0002921.

Convective drying kinetics of strawberry (Fragaria ananassa): Effects on antioxidant activity, anthocyanins and total phenolic content.

PubMed

Méndez-Lagunas, Lilia; Rodríguez-Ramírez, Juan; Cruz-Gracida, Marlene; Sandoval-Torres, Sadoth; Barriada-Bernal, Gerardo

2017-09-01

The thermal drying effects on strawberries were investigated in terms of the kinetics of antioxidant activity (AA), anthocyanins (A) and total phenolic compound content (TPC), as well as the final colour. The evaluated drying temperatures were 50 and 60°C with an air rate of 1.5m/s. The 2,2-diphenyl-2-picryl-hydrazyl, pH differential and Folin-Ciocalteu methods were used to assess the antioxidant properties. The kinetics of TPC and AA showed an initial and final period of degradation attributed to inhibition of enzymes. A plateau between these two periods suggests that under certain conditions of temperature and water content, no degradation reactions occurred. Final losses of up to 74, 45 and 78% were found for AA, A and TPC, respectively. The total colour change (ΔE) was lesser degree at 60 than 50°C. Thermal degradation of the antioxidant compounds followed a first-order reaction kinetics and the degradation rate constants (k) were calculated. Copyright © 2017 Elsevier Ltd. All rights reserved.

Response surface methodology for evaluation and optimization of process parameter and antioxidant capacity of rice flour modified by enzymatic extrusion.

PubMed

Xu, Enbo; Pan, Xiaowei; Wu, Zhengzong; Long, Jie; Li, Jingpeng; Xu, Xueming; Jin, Zhengyu; Jiao, Aiquan

2016-12-01

For the purpose of investigating the effect of enzyme concentration (EC), barrel temperature (BT), moisture content (MC), and screw speed (SS) on processing parameters (product temperature, die pressure and special mechanical energy (SME)) and product responses (extent of gelatinization (GE), retention rate of total phenolic content (TPC-RR)), rice flour extruded with thermostable α-amylase was analyzed by response surface methodology. Stepwise regression models were computed to generate response surface and contour plots, revealing that both TPC-RR and GE increased as increasing MC while expressed different sensitivities to BT during enzymatic extrusion. Phenolics preservation was benefited from low SME. According to multiple-factor optimization, the conditions required to obtain the target SME (10kJ/kg), GE (100%) and TPC-RR (85%) were: EC=1.37‰, BT=93.01°C, MC=44.30%, and SS=171.66rpm, with the actual values (9.49kJ/kg, 99.96% and 87.10%, respectively) showing a good fit to the predicted values. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

3D simulation of electron and ion transmission of GEM-based detectors

NASA Astrophysics Data System (ADS)

Bhattacharya, Purba; Mohanty, Bedangadas; Mukhopadhyay, Supratik; Majumdar, Nayana; da Luz, Hugo Natal

2017-10-01

Time Projection Chamber (TPC) has been chosen as the main tracking system in several high-flux and high repetition rate experiments. These include on-going experiments such as ALICE and future experiments such as PANDA at FAIR and ILC. Different R&D activities were carried out on the adoption of Gas Electron Multiplier (GEM) as the gas amplification stage of the ALICE-TPC upgrade version. The requirement of low ion feedback has been established through these activities. Low ion feedback minimizes distortions due to space charge and maintains the necessary values of detector gain and energy resolution. In the present work, Garfield simulation framework has been used to study the related physical processes occurring within single, triple and quadruple GEM detectors. Ion backflow and electron transmission of quadruple GEMs, made up of foils with different hole pitch under different electromagnetic field configurations (the projected solutions for the ALICE TPC) have been studied. Finally a new triple GEM detector configuration with low ion backflow fraction and good electron transmission properties has been proposed as a simpler GEM-based alternative suitable for TPCs for future collider experiments.

Development of a 3-D Nuclear Event Visualization Program Using Unity

NASA Astrophysics Data System (ADS)

Kuhn, Victoria

2017-09-01

Simulations have become increasingly important for science and there is an increasing emphasis on the visualization of simulations within a Virtual Reality (VR) environment. Our group is exploring this capability as a visualization tool not just for those curious about science, but also for educational purposes for K-12 students. Using data collected in 3-D by a Time Projection Chamber (TPC), we are able to visualize nuclear and cosmic events. The Unity game engine was used to recreate the TPC to visualize these events and construct a VR application. The methods used to create these simulations will be presented along with an example of a simulation. I will also present on the development and testing of this program, which I carried out this past summer at MSU as part of an REU program. We used data from the S πRIT TPC, but the software can be applied to other 3-D detectors. This work is supported by the U.S. Department of Energy under Grant Nos. DE-SC0014530, DE-NA0002923 and US NSF under Grant No. PHY-1565546.

The Active Target Time Projection Chamber at NSCL

NASA Astrophysics Data System (ADS)

Bazin, D.; Bradt, J.; Ayyad, Y.; Mittig, W.; Ahn, T.; Beceiro-Novo, S.; Carpenter, L.; Cortesi, M.; Fritsch, A.; Kolata, J. J.; Lynch, W.; Watwood, N.

2017-11-01

Reactions in inverse kinematics close to the Coulomb barrier offer unique opportunities to study exotic nuclei, but they are plagued by the difficulty to efficiently and precisely measure the characteristics of the emerging particles. The Active Target Time Projection Chamber (AT-TPC) offers an elegant solution to this dilemma. In this device, the detector gas of the time projection chamber is at the same time the target in which nuclear reactions take place. The use of this new paradigm offers several advantages over conventional inert target methods, the most significant being the ability to increase the luminosity of experiments without loss of resolution. The AT-TPC and some results obtained on resonant α scattering to explore the clustering properties of neutron-rich nuclei are presented, as well as fusion cross section results using a 10Be radioactive beam. In addition, the first re-accelerated radioactive beam experiment using the fully commissioned ReA3 linac was conducted recently at the NSCL with the AT-TPC, where proton resonant scattering of a 4.6 MeV/u 46Ar beam was used to measure the neutron single-particle strength in 47Ar.

Exploring 0.1-10 eV axions with a new helioscope concept

NASA Astrophysics Data System (ADS)

Galán, J.; Dafni, T.; Ferrer-Ribas, E.; Giomataris, I.; Iguaz, F. J.; Irastorza, I. G.; García, J. A.; Garza, J. G.; Luzon, G.; Papaevangelou, T.; Redondo, J.; Tomás, A.

2015-12-01

We explore the possibility to develop a new axion helioscope type, sensitive to the higher axion mass region favored by axion models. We propose to use a low background large volume TPC immersed in an intense magnetic field. Contrary to traditional tracking helioscopes, this detection technique takes advantage of the capability to directly detect the photons converted on the buffer gas which defines the axion mass sensitivity region, and does not require pointing the magnet to the Sun. The operation flexibility of a TPC to be used with different gas mixtures (He, Ne, Xe, etc.) and pressures (from 10 mbar to 10 bar) will allow to enhance sensitivity for axion masses from few meV to several eV. We present different helioscope data taking scenarios, considering detection efficiency and axion absorption probability, and show the sensitivities reachable with this technique to be few × 10-11 GeV-1 for a 5 T, m3 scale TPC. We show that a few years program taking data with such setup would allow to probe the KSVZ axion model for axion masses above 0gtrsim 10 meV.

Effects of co-fermentation by Saccharomyces cerevisiae and Issatchenkia orientalis on sea buckthorn juice.

PubMed

Negi, Bharti; Dey, Gargi

2013-06-01

This work relates to the development of a co-fermented product of sea buckthorn (Hippophae rhamnoides L.) with Saccharomyces cerevisiae and Issatchenkia orientalis. Besides malic acid degradation, the parameters of present production technology were also standardized with emphasis on the retainability of total phenolic content (TPC) of sea buckthorn juice. The effect of co-fermentation on physico-chemical characteristics, organic acids, flavonoids, TPC and antioxidant activities was studied. The high-performance liquid chromatography (HPLC) analysis showed 55% reduction in malic acid content after the co-fermentation of sea buckthorn juice. The TPC of sea buckthorn product was 2.18 g gallic acid equivalent (GAE)/l. The estimated scavenging effect on 2,2-diphenyl-1-picrylhydrazyl free radicals was 2.63 Trolox equivalent (TE) mmol/l. Ferric-reducing antioxidant power and 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulphonic acid) assays also showed that sea buckthorn product was on a par with commercial wines (Cabernet Shiraz and Beaujolais). We conclude that the process of co-fermentation resulted in a significant antioxidant potential of sea buckthorn product.

Extraction kinetic modelling of total polyphenols and total anthocyanins from saffron floral bio-residues: Comparison of extraction methods.

PubMed

Da Porto, Carla; Natolino, Andrea

2018-08-30

Analysis of the extraction kinetic modelling for natural compounds is essential for industrial application. The second order rate model was applied to estimate the extraction kinetics of conventional solid-liquid extraction (CSLE), ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) of total polyphenols (TPC) from saffron floral bio-residues at different solid-to-liquid ratios (R S/L )(1:10, 1:20, 1:30, 1:50 g ml -1 ), ethanol 59% as solvent and 66 °C temperature. The optimum solid-to-liquid ratios for TPC kinetics were 1:20 for CLSE, 1:30 for UAE and 1:50 for MAE. The kinetics of total anthocyanins (TA) and antioxidant activity (AA) were investigated for the optimum R S/L for each method. The results showed a good prediction of the model for extraction kinetics in all experiments (R 2  > 0.99; NRMS 0.65-3.35%). The kinetic parameters were calculated and discussed. UAE, compared with the other methods, had the greater efficiency for TPC, TA and AA. Copyright © 2018 Elsevier Ltd. All rights reserved.

Measurement of the normalized 238U(n ,f )/235U(n ,f ) cross section ratio from threshold to 30 MeV with the NIFFTE fission Time Projection Chamber

NASA Astrophysics Data System (ADS)

Casperson, R. J.; Asner, D. M.; Baker, J.; Baker, R. G.; Barrett, J. S.; Bowden, N. S.; Brune, C.; Bundgaard, J.; Burgett, E.; Cebra, D. A.; Classen, T.; Cunningham, M.; Deaven, J.; Duke, D. L.; Ferguson, I.; Gearhart, J.; Geppert-Kleinrath, V.; Greife, U.; Grimes, S.; Guardincerri, E.; Hager, U.; Hagmann, C.; Heffner, M.; Hensle, D.; Hertel, N.; Higgins, D.; Hill, T.; Isenhower, L. D.; King, J.; Klay, J. L.; Kornilov, N.; Kudo, R.; Laptev, A. B.; Loveland, W.; Lynch, M.; Lynn, W. S.; Magee, J. A.; Manning, B.; Massey, T. N.; McGrath, C.; Meharchand, R.; Mendenhall, M. P.; Montoya, L.; Pickle, N. T.; Qu, H.; Ruz, J.; Sangiorgio, S.; Schmitt, K. T.; Seilhan, B.; Sharma, S.; Snyder, L.; Stave, S.; Tate, A. C.; Tatishvili, G.; Thornton, R. T.; Tovesson, F.; Towell, D. E.; Towell, R. S.; Walsh, N.; Watson, S.; Wendt, B.; Wood, L.; Yao, L.; Younes, W.; Niffte Collaboration

2018-03-01

The normalized 238U(n ,f )/235U(n ,f ) cross section ratio has been measured using the NIFFTE fission Time Projection Chamber (fissionTPC) from the reaction threshold to 30 MeV . The fissionTPC is a two-volume MICROMEGAS time projection chamber that allows for full three-dimensional reconstruction of fission-fragment ionization profiles from neutron-induced fission. The measurement was performed at the Los Alamos Neutron Science Center, where the neutron energy is determined from neutron time of-flight. The 238U(n ,f )/235U(n ,f ) ratio reported here is the first cross section measurement made with the fissionTPC, and will provide new experimental data for evaluation of the 238U(n ,f ) cross section, an important standard used in neutron-flux measurements. Use of a development target in this work prevented the determination of an absolute normalization, to be addressed in future measurements. Instead, the measured cross section ratio has been normalized to ENDF/B-VIII.β 5 at 14.5 MeV.

Characterization of free, conjugated and bound phenolics and lipophilic antioxidants in regular- and non-darkening cranberry beans (Phaseolus vulgaris L.).

PubMed

Chen, Peter X; Tang, Yao; Marcone, Massimo F; Pauls, Peter K; Zhang, Bing; Liu, Ronghua; Tsao, Rong

2015-10-15

Cranberry beans (Phaseolus vulgaris L.) from 7 different cultivars were characterized for phytochemicals and assessed for antioxidant activities. In vitro colorimetric methods were used to measure total phenolic (TPC) and total proanthocyanidin (PAC) contents. Free, conjugated and bound phenolic acids and flavonoids were also identified and quantified using HPLC-DAD/ESI-MS(n). Regular-darkening (RD) seeds contained higher TPC, PAC and flavonoids which were absent in the non-darkening (ND) seeds. Bound and conjugated phenolics in RD and ND mainly included cinnamic and benzoic acids. DPPH, FRAP and ORAC showed strong positive correlation with TPC, PAC, and with specific phenolics such as free catechin and bound p-hydroxybenzoic acid. Lipophilic extracts were rich in polyunsaturated fatty acids (69.20-76.89%). Carotenoid and tocopherol were limited to γ-tocopherol and β-carotene. Results from this study can contribute to the development of cranberry bean cultivars with increased health benefits and addresses specific phenolic contributors to antioxidant activity. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

Change of Bioactive Constituent in Clinacanthus nutans Leaves under Sun Drying

NASA Astrophysics Data System (ADS)

Abdullah, Sriyana; Aziz, Muhamad Faris Abdul

2018-03-01

Clinacanthus nutans (C. nutans) or locally known as belalai gajah is a folk medicine since ancient time. This research project was established to investigate the effects of under sun drying on the C. nutans bioactive constituent. The drying experiments were conducted using different drying surfaces i.e. perforated, black polythene and white polythene. The fresh and dried leaves were then extracted using a sonicator to evaluate its bioactive constituent. The total phenolic content (TPC) in the C. nutans extracts were determined using Follin Ciocalteu reagent method to represent the bioactive constituent. Drying over the white polythene surface showed the slowest reduction of moisture content as compared to the perforated polythene and black surfaces. Results also showed no significant effect of the drying surfaces on the TPC. However, the TPC in the dried leaves was significantly higher than in the fresh leaves. This may be due to the plant cells response to abiotic stress and the inhibition of oxidation enzymes. Therefore, drying C. nutanc leaves under sun light could be considered in order to preserve the concentration of phenolic compounds and for minimizing energy consumption.

Tau Kinetics in Neurons and the Human Central Nervous System.

PubMed

Sato, Chihiro; Barthélemy, Nicolas R; Mawuenyega, Kwasi G; Patterson, Bruce W; Gordon, Brian A; Jockel-Balsarotti, Jennifer; Sullivan, Melissa; Crisp, Matthew J; Kasten, Tom; Kirmess, Kristopher M; Kanaan, Nicholas M; Yarasheski, Kevin E; Baker-Nigh, Alaina; Benzinger, Tammie L S; Miller, Timothy M; Karch, Celeste M; Bateman, Randall J

2018-03-21

We developed stable isotope labeling and mass spectrometry approaches to measure the kinetics of multiple isoforms and fragments of tau in the human central nervous system (CNS) and in human induced pluripotent stem cell (iPSC)-derived neurons. Newly synthesized tau is truncated and released from human neurons in 3 days. Although most tau proteins have similar turnover, 4R tau isoforms and phosphorylated forms of tau exhibit faster turnover rates, suggesting unique processing of these forms that may have independent biological activities. The half-life of tau in control human iPSC-derived neurons is 6.74 ± 0.45 days and in human CNS is 23 ± 6.4 days. In cognitively normal and Alzheimer's disease participants, the production rate of tau positively correlates with the amount of amyloid plaques, indicating a biological link between amyloid plaques and tau physiology. Copyright © 2018 Elsevier Inc. All rights reserved.

Compartmentalized PDE4A5 Signaling Impairs Hippocampal Synaptic Plasticity and Long-Term Memory.

PubMed

Havekes, Robbert; Park, Alan J; Tolentino, Rosa E; Bruinenberg, Vibeke M; Tudor, Jennifer C; Lee, Yool; Hansen, Rolf T; Guercio, Leonardo A; Linton, Edward; Neves-Zaph, Susana R; Meerlo, Peter; Baillie, George S; Houslay, Miles D; Abel, Ted

2016-08-24

Alterations in cAMP signaling are thought to contribute to neurocognitive and neuropsychiatric disorders. Members of the cAMP-specific phosphodiesterase 4 (PDE4) family, which contains >25 different isoforms, play a key role in determining spatial cAMP degradation so as to orchestrate compartmentalized cAMP signaling in cells. Each isoform binds to a different set of protein complexes through its unique N-terminal domain, thereby leading to targeted degradation of cAMP in specific intracellular compartments. However, the functional role of specific compartmentalized PDE4 isoforms has not been examined in vivo Here, we show that increasing protein levels of the PDE4A5 isoform in mouse hippocampal excitatory neurons impairs a long-lasting form of hippocampal synaptic plasticity and attenuates hippocampus-dependent long-term memories without affecting anxiety. In contrast, viral expression of a truncated version of PDE4A5, which lacks the unique N-terminal targeting domain, does not affect long-term memory. Further, overexpression of the PDE4A1 isoform, which targets a different subset of signalosomes, leaves memory undisturbed. Fluorescence resonance energy transfer sensor-based cAMP measurements reveal that the full-length PDE4A5, in contrast to the truncated form, hampers forskolin-mediated increases in neuronal cAMP levels. Our study indicates that the unique N-terminal localization domain of PDE4A5 is essential for the targeting of specific cAMP-dependent signaling underlying synaptic plasticity and memory. The development of compounds to disrupt the compartmentalization of individual PDE4 isoforms by targeting their unique N-terminal domains may provide a fruitful approach to prevent cognitive deficits in neuropsychiatric and neurocognitive disorders that are associated with alterations in cAMP signaling. Neurons exhibit localized signaling processes that enable biochemical cascades to be activated selectively in specific subcellular compartments. The phosphodiesterase 4 (PDE4) family coordinates the degradation of cAMP, leading to the local attenuation of cAMP-dependent signaling pathways. Sleep deprivation leads to increased hippocampal expression of the PDE4A5 isoform. Here, we explored whether PDE4A5 overexpression mimics behavioral and synaptic plasticity phenotypes associated with sleep deprivation. Viral expression of PDE4A5 in hippocampal neurons impairs long-term potentiation and attenuates the formation of hippocampus-dependent long-term memories. Our findings suggest that PDE4A5 is a molecular constraint on cognitive processes and may contribute to the development of novel therapeutic approaches to prevent cognitive deficits in neuropsychiatric and neurocognitive disorders that are associated with alterations in cAMP signaling. Copyright © 2016 Havekes et al.

The bioartificial thyroid: a biotechnological perspective in endocrine organ engineering for transplantation replacement.

PubMed

Toni, Roberto; Casa, Claudia Della; Spaletta, Giulia; Marchetti, Giacomo; Mazzoni, Perseo; Bodria, Monica; Ravera, Simone; Dallatana, Davide; Castorina, Sergio; Riccioli, Vincenzo; Castorina, Emilio Giovanni; Antoci, Salvatore; Campanile, Enrico; Raise, Gabriella; Scalise, Gabriella; Rossi, Raffaella; Rossio, Raffaella; Ugolotti, Giorgio; Ugolottio, Giorgio; Martorella, Andrew; Roti, Elio; Rot, Elio; Sgallari, Fiorella; Pinchera, Aldo

2007-01-01

A new concept for ex situ endocrine organ bioengineering is presented, focused on the realization of a human bioartificial thyroid gland. It is based on the theoretical assumption and experimental evidence that symmetries in geometrical coordinates of the thyroid tissue remain invariant with respect to developmental, physiological or pathophysiological transformations occuring in the gland architecture. This topological arrangement is dependent upon physical connections established between cells, cell adhesion molecules and extracellular matrix, leading to the view that the thyroid parenchyma behaves like a deformable "putty", moulded onto an elastic stromal/vascular scaffold (SVS) dictating the final morphology of the gland. In particular, we have raised the idea that the geometry of the SVS per se provides pivotal epigenetic information to address the genetically-programmed, thyrocyte and endothelial/vascular proliferation and differentiation towards a functionally mature gland, making organ form a pre-requirementfor organ function. A number of experimental approaches are explored to obtain a reliable replica of a human thyroid SVS, and an informatic simulation is designed based on fractal growth of the thyroid intraparenchymal arterial tree. Various tissue-compatible and degradable synthetic or biomimetic polymers are discussed to act as a template of the thyroid SVS, onto which to co-seed autologous human thyrocyte (TPC) and endothelial/vascular (EVPC) progenitor cells. Harvest and expansion of both TPC and EVPC in primary culture are considered, with specific attention to the selection of normal thyrocytes growing at a satisfactory rate to colonize the synthetic matrix. In addition, both in vitro and in vivo techniques to authenticate TPC and EVPC lineage differentiation are reviewed, including immunocytochemistry, reverse trascriptase-polymerase chain reaction, flow cytomery and proteomics. Finally, analysis of viability of the thyroid construct following implantation in animal hosts is proposed, with the intent to obtain a bioartificial thyroid gland morphologically and functionally adequate for transplantation. We believe that the biotechnological scenario proposed herein may provide a template to construct other, more complex and clinically-relevant bioartificial endocrine organs ex situ, such as human pancreatic islets and the liver, and perhaps a new approach to brain bioengineering.

Different KChIPs compete for heteromultimeric assembly with pore-forming Kv4 subunits.

PubMed

Zhou, Jingheng; Tang, Yiquan; Zheng, Qin; Li, Meng; Yuan, Tianyi; Chen, Liangyi; Huang, Zhuo; Wang, KeWei

2015-06-02

Auxiliary Kv channel-interacting proteins 1-4 (KChIPs1-4) coassemble with pore-forming Kv4 α-subunits to form channel complexes underlying somatodendritic subthreshold A-type current that regulates neuronal excitability. It has been hypothesized that different KChIPs can competitively bind to Kv4 α-subunit to form variable channel complexes that can exhibit distinct biophysical properties for modulation of neural function. In this study, we use single-molecule subunit counting by total internal reflection fluorescence microscopy in combinations with electrophysiology and biochemistry to investigate whether different isoforms of auxiliary KChIPs, KChIP4a, and KChIP4bl, can compete for binding of Kv4.3 to coassemble heteromultimeric channel complexes for modulation of channel function. To count the number of photobleaching steps solely from cell membrane, we take advantage of a membrane tethered k-ras-CAAX peptide that anchors cytosolic KChIP4 proteins to the surface for reduction of background noise. Single-molecule subunit counting reveals that the number of KChIP4 isoforms in Kv4.3-KChIP4 complexes can vary depending on the KChIP4 expression level. Increasing the amount of KChIP4bl gradually reduces bleaching steps of KChIP4a isoform proteins, and vice versa. Further analysis of channel gating kinetics from different Kv4-KChIP4 subunit compositions confirms that both KChIP4a and KChIP4bl can modulate the channel complex function upon coassembly. Taken together, our findings show that auxiliary KChIPs can heteroassemble with Kv4 in a competitive manner to form heteromultimeric Kv4-KChIP4 channel complexes that are biophysically distinct and regulated under physiological or pathological conditions. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Antioxidant compounds of kiwifruit during post-ripening process at ambient temperature

NASA Astrophysics Data System (ADS)

Liang, D.; Lv, X. L.; Wang, J.; Xia, H.; Xie, Y.; Li, M. Z.; Wang, Y. Z.

2017-09-01

Kiwifruit is well-known for an excellent source of antioxidants. In this study, contents of total phenolics (TPC), total flavonoids (TFC), total flavanols (TFAC) and vitamin C were investigated in different fruit tissues during post-ripening process at ambient temperature. The results explored that TPC and TFC showed declining trend with the increase in storage interval in different tissues. TFAC raised with the increase in storage interval in different fruit tissues, while was followed a decrease in later process. Vitamin C content was stable in outer and inner pericarp in prometaphase of post-ripening.

Modeling Electronegative Impurity Concentrations in Liquid Argon Detectors

NASA Astrophysics Data System (ADS)

Tang, Wei; Li, Yichen; Thorn, Craig; Qian, Xin

2017-01-01

Achieving long electron lifetime is crucial to reach the high performance of large Liquid Argon Time Projection Chamber (LArTPC) envisioned for next generation neutrino experiments. We have built up a quantitative model to describe the impurity distribution and transportation in a cryostat. Henrys constants of Oxygen and water, which describe the partition of impurities between gas argon and liquid argon, have been deduced through this model with the measurements in BNL 20-L LAr test stand. These results indicate the importance of the gas purification system and prospects on large LArTPC detectors will be discussed.

Front-end electronics for the LZ experiment

NASA Astrophysics Data System (ADS)

Morad, James; LZ Collaboration

2016-03-01

LZ is a second generation direct dark matter detection experiment with 5.6 tonnes of liquid xenon active target, which will be instrumented as a two-phase time projection chamber (TPC). The peripheral xenon outside the active TPC (``skin'') will also be instrumented. In addition, there will be a liquid scintillator based outer veto surrounding the main cryostat. All of these systems will be read out using photomultiplier tubes. I will present the designs for front-end electronics for all these systems, which have been optimized for shaping times, gains, and low noise. Preliminary results from prototype boards will also be presented.

The Origins and Evolution of the Time Projection Chamber (TPC) Idea

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nygren, David

In February 1974, I conceived an idea for a tracking detector with only one spatial projection, thereby eliminating ambiguities that occur in conventional detector systems based on wires. I called it the “Time Projection Chamber”, or TPC, a name that has stuck even though the concept has evolved considerably over the following decades. I will recount the history leading to its conception and development in that now distant epoch, and will attempt to show why this is an interesting and instructive story and how the idea may continue to extend scientific reach in the coming era.

Characterisation of phenol oxidase and peroxidase from maize silk.

PubMed

Sukalović, V Hadzi-Tasković; Veljović-Jovanović, S; Maksimović, J Dragisić; Maksimović, V; Pajić, Z

2010-05-01

Silk of some maize genotypes contains a high level of phenolics that undergo enzymatic oxidation to form quinones, which condense among themselves or with proteins to form brown pigments. Two phenolic oxidizing enzymes, peroxidase (POD; EC 1.11.1.7) and polyphenol oxidase (PPO; EC 1.10.3.1), from maize (Zea mays L.) silk were characterised with respect to their preferred substrate, different isoforms and specific effectors. One browning silk sample with high, and two non-browning samples with low phenolic content were investigated. Although POD oxidizes a wide range of phenolic substrates in vitro, its activity rate was independent of silk phenolic content. PPO activity, detected with o-diphenolic substrates, was abundant only in browning silk, and low or absent in non-browning silk. Pollination increased POD but not PPO activity. Isoelectric-focusing (IEF) and specific staining for POD and PPO showed a high degree of polymorphism that varied with silk origin. The IEF pattern of POD revealed a number of anionic and several cationic isoenzymes, with the most pronounced having neutral pI 7 and a basic isoform with pI 10. Detected isoforms of PPO were anionic, except for one neutral form found only in browning silk, and occupied positions different from those of POD. Different inhibitory effects of NaN(3), EDTA, KCN, and L-cysteine, as well as different impacts of a variety of cations on the oxidation of chlorogenic acid, mediated by PPO or POD, were detected. The findings are discussed in terms of a possible roles of these enzymes in defence and pollination.

Beta 1D integrin displaces the beta 1A isoform in striated muscles: localization at junctional structures and signaling potential in nonmuscle cells.

PubMed

Belkin, A M; Zhidkova, N I; Balzac, F; Altruda, F; Tomatis, D; Maier, A; Tarone, G; Koteliansky, V E; Burridge, K

1996-01-01

The cytoplasmic domains of integrins provide attachment of these extracellular matrix receptors to the cytoskeleton and play a critical role in integrin-mediated signal transduction. In this report we describe the identification, expression, localization, and initial functional characterization of a novel form of beta 1 integrin, termed beta 1D. This isoform contains a unique alternatively spliced cytoplasmic domain of 50 amino acids, with the last 24 amino acids encoded by an additional exon. Of these 24 amino acids, 11 are conserved when compared to the beta 1A isoform, but 13 are unique (Zhidkova, N. I., A. M. Belkin, and R. Mayne. 1995. Biochem. Biophys. Res. Commun. 214:279-285; van der Flier, A., I. Kuikman, C. Baudoin, R, van der Neuf, and A. Sonnenberg. 1995. FEBS Lett. 369:340-344). Using an anti-peptide antibody against the beta 1D integrin subunit, we demonstrated that the beta 1D isoform is synthesized only in skeletal and cardiac muscles, while very low amounts of beta 1A were detected by immunoblot in striated muscles. Whereas beta 1A could not be detected in adult skeletal muscle fibers and cardiomyocytes by immunofluorescence, beta 1D was localized to the sarcolemma of both cell types. In skeletal muscle, beta 1D was concentrated in costameres, myotendinous, and neuromuscular junctions. In cardiac muscle this beta 1 isoform was found in costamers and intercalated discs. beta 1D was associated with alpha 7A and alpha 7B in adult skeletal muscle. In cardiomyocytes of adult heart, alpha 7B was the major partner for the beta 1D isoform. beta 1D could not be detected in proliferating C2C12 myoblasts, but it appeared immediately after myoblast fusion and its amount continued to rise during myotube growth and maturation. In contrast, expression of the beta 1A isoform was downregulated during myodifferentiation in culture and it was completely displaced by beta 1D in mature differentiated myotubes. We also analyzed some functional properties of the beta 1D integrin subunit. Expression of human beta 1D in CHO cells led to its localization at focal adhesions. Clustering of this integrin isoform on the cell surface stimulated tyrosine phosphorylation of pp125FAK (focal adhesion kinase) and caused transient activation of mitogen-activated protein (MAP) kinases. These data indicate that beta 1D and beta 1A integrin isoforms are functionally similar with regard to integrin-mediated signaling.

Isoforms of Vitamin E Differentially Regulate PKC α and Inflammation: A Review

PubMed Central

Cook-Mills, Joan M.

2013-01-01

Vitamin E regulation of disease has been extensively studied but most studies focus on the α-tocopherol isoform of vitamin E. These reports indicate contradictory outcomes for anti-inflammatory functions of the α-tocopherol isoform of vitamin E with regards to animal and clinical studies. These seemingly disparate results are consistent with our recent studies demonstrating that purified natural forms of vitamin E have opposing regulatory functions during inflammation. In this review, we discuss that α-tocopherol inhibits whereas γ-tocopherol elevates allergic inflammation, airway hyperresponsiveness, leukocyte transendothelial migration, and endothelial cell adhesion molecule signaling through protein kinase Cα. Moreover, we have demonstrated that α-tocopherol is an antagonist and γ-tocopherol is an agonist of PKCα through direct binding to a regulatory domain of PKCα. In summary, we have determined mechanisms for opposing regulatory functions of α-tocopherol and γ-tocopherol on inflammation. Information from our studies will have significant impact on the design of clinical studies and on vitamin E consumption. PMID:23977443

Identification of two frataxin isoforms in Zea mays: Structural and functional studies.

PubMed

Buchensky, Celeste; Sánchez, Manuel; Carrillo, Martin; Palacios, Oscar; Capdevila, Mercè; Domínguez-Vera, Jose M; Busi, Maria V; Atrian, Sílvia; Pagani, Maria A; Gomez-Casati, Diego F

2017-09-01

Frataxin is a ubiquitous protein that plays a role in Fe-S cluster biosynthesis and iron and heme metabolism, although its molecular functions are not entirely clear. In non-photosynthetic eukaryotes, frataxin is encoded by a single gene, and the protein localizes to mitochondria. Here we report the presence of two functional frataxin isoforms in Zea mays, ZmFH-1 and ZmFH-2. We confirmed our previous findings regarding plant frataxins: both proteins have dual localization in mitochondria and chloroplasts. Physiological, biochemical and biophysical studies show some differences in the expression pattern, protection against oxidants and in the aggregation state of both isoforms, suggesting that the two frataxin homologs would play similar but not identical roles in plant cell metabolism. In addition, two specific features of plant frataxins were evidenced: their ability to form dimers and their tendency to undergo conformational change under oxygen exposure. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

A Novel Isoform of Sucrose Synthase Is Targeted to the Cell Wall during Secondary Cell Wall Synthesis in Cotton Fiber[C][W][OA

PubMed Central

Brill, Elizabeth; van Thournout, Michel; White, Rosemary G.; Llewellyn, Danny; Campbell, Peter M.; Engelen, Steven; Ruan, Yong-Ling; Arioli, Tony; Furbank, Robert T.

2011-01-01

Sucrose (Suc) synthase (Sus) is the major enzyme of Suc breakdown for cellulose biosynthesis in cotton (Gossypium hirsutum) fiber, an important source of fiber for the textile industry. This study examines the tissue-specific expression, relative abundance, and temporal expression of various Sus transcripts and proteins present in cotton. A novel isoform of Sus (SusC) is identified that is expressed at high levels during secondary cell wall synthesis in fiber and is present in the cell wall fraction. The phylogenetic relationships of the deduced amino acid sequences indicate two ancestral groups of Sus proteins predating the divergence of monocots and dicots and that SusC sequences form a distinct branch in the phylogeny within the dicot-specific clade. The subcellular location of the Sus isoforms is determined, and it is proposed that cell wall-localized SusC may provide UDP-glucose for cellulose and callose synthesis from extracellular sugars. PMID:21757635

Analysis of human articular chondrocyte CD44 isoform expression and function in health and disease.

PubMed

Salter, D M; Godolphin, J L; Gourlay, M S; Lawson, M F; Hughes, D E; Dunne, E

1996-08-01

Interactions between articular chondrocytes and components of the extracellular matrix are of potential importance in the normal function of cartilage and in the pathophysiology of arthritis. Little is known of the basis of these interactions, but cell adhesive molecules such as CD44 are likely to be involved. Immunohistology using six well-characterized anti-CD44 monoclonal antibodies demonstrated standard CD44 isoform (CD44H) expression by all chondrocytes in normal and osteoarthrotic (OA) cartilage but absence of the CD44E variant. Polymerase chain reaction (PCR) of reverse transcribed mRNA from monolayer cultures of normal and OA chondrocytes using primer sequences which span the region containing variably spliced exons produced a predominant band representing the standard form of CD44, which lacks the variable exons 6-15 (v1-v10). No product was seen at the expected size of the epithelial variant of CD44 (CD44v8-10). Use of exon-specific primers, however, showed expression of variant exons resulting in multiple minor isoforms. Standard CD44 was also shown to be the predominantly expressed isoform identified by immunoprecipitation, but human articular chondrocytes did not adhere to hyaluronan in vitro. Chondrocyte CD44 may function as an adhesion receptor for other matrix molecules such as fibronectin or collagen.

Kinetics and structural features of dimeric glutamine-dependent bacterial NAD+ synthetases suggest evolutionary adaptation to available metabolites.

PubMed

Santos, Adrian Richard Schenberger; Gerhardt, Edileusa Cristina Marques; Moure, Vivian Rotuno; Pedrosa, Fábio Oliveira; Souza, Emanuel Maltempi; Diamanti, Riccardo; Högbom, Martin; Huergo, Luciano Fernandes

2018-05-11

NADH (NAD + ) and its reduced form NADH serve as cofactors for a variety of oxidoreductases that participate in many metabolic pathways. NAD + also is used as substrate by ADP-ribosyl transferases and by sirtuins. NAD + biosynthesis is one of the most fundamental biochemical pathways in nature, and the ubiquitous NAD + synthetase (NadE) catalyzes the final step in this biosynthetic route. Two different classes of NadE have been described to date: dimeric single-domain ammonium-dependent NadE NH3 and octameric glutamine-dependent NadE Gln , and the presence of multiple NadE isoforms is relatively common in prokaryotes. Here, we identified a novel dimeric group of NadE Gln in bacteria. Substrate preferences and structural analyses suggested that dimeric NadE Gln enzymes may constitute evolutionary intermediates between dimeric NadE NH3 and octameric NadE Gln The characterization of additional NadE isoforms in the diazotrophic bacterium Azospirillum brasilense along with the determination of intracellular glutamine levels in response to an ammonium shock led us to propose a model in which these different NadE isoforms became active accordingly to the availability of nitrogen. These data may explain the selective pressures that support the coexistence of multiple isoforms of NadE in some prokaryotes. © 2018 Santos et al.

Crystallization and preliminary X-ray crystallographic analysis of latent isoform PPO4 mushroom (Agaricus bisporus) tyrosinase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mauracher, Stephan Gerhard; Molitor, Christian; Al-Oweini, Rami

Polyphenol oxidase 4 (PPO4) from the natural source A. bisporus was crystallized in its latent precursor form (pro-tyrosinase; Ser2–Thr565) using the 6-tungstotellurate(VI) salt Na{sub 6}[TeW{sub 6}O{sub 24}]·22H{sub 2}O as a crystallization additive. Tyrosinase exhibits catalytic activity for the ortho-hydroxylation of monophenols to diphenols as well as their subsequent oxidation to quinones. Owing to polymerization of these quinones, brown-coloured high-molecular-weight compounds called melanins are generated. The latent precursor form of polyphenol oxidase 4, one of the six tyrosinase isoforms from Agaricus bisporus, was purified to homogeneity and crystallized. The obtained crystals belonged to space group C121 (two molecules per asymmetric unit)more » and diffracted to 2.78 Å resolution. The protein only formed crystals under low-salt conditions using the 6-tungstotellurate(VI) salt Na{sub 6}[TeW{sub 6}O{sub 24}]·22H{sub 2}O as a co-crystallization agent.« less

Reduced peripheral expression of the glucocorticoid receptor α isoform in individuals with posttraumatic stress disorder: a cumulative effect of trauma burden.

PubMed

Gola, Hannah; Engler, Andrea; Morath, Julia; Adenauer, Hannah; Elbert, Thomas; Kolassa, Iris-Tatjana; Engler, Harald

2014-01-01

Posttraumatic stress disorder (PTSD) is a serious psychiatric condition that was found to be associated with altered functioning of the hypothalamic-pituitary-adrenal (HPA) axis and changes in glucocorticoid (GC) responsiveness. The physiological actions of GCs are primarily mediated through GC receptors (GR) of which isoforms with different biological activities exist. This study aimed to investigate whether trauma-experience and/or PTSD are associated with altered expression of GR splice variants. GRα and GRβ mRNA expression levels were determined by real-time quantitative PCR in whole blood samples of individuals with chronic and severe forms of PTSD (n = 42) as well as in ethnically matched reference subjects (non-PTSD, n = 35). Individuals suffering from PTSD exhibited significantly lower expression of the predominant and functionally active GRα isoform compared to non-PTSD subjects. This effect remained significant when accounting for gender, smoking, psychotropic medication or comorbid depression. Moreover, the GRα expression level was significantly negatively correlated with the number of traumatic event types experienced, both in the whole sample and within the PTSD patient group. Expression of the less abundant and non-ligand binding GRβ isoform was comparable between patient and reference groups. Reduced expression of the functionally active GRα isoform in peripheral blood cells of individuals with PTSD seems to be a cumulative effect of trauma burden rather than a specific feature of PTSD since non-PTSD subjects with high trauma load showed an intermediate phenotype between PTSD patients and individuals with no or few traumatic experiences.

Temporal expression and mitochondrial localization of a Foxp2 isoform lacking the forkhead domain in developing Purkinje cells.

PubMed

Tanabe, Yuko; Fujiwara, Yuji; Matsuzaki, Ayumi; Fujita, Eriko; Kasahara, Tadashi; Yuasa, Shigeki; Momoi, Takashi

2012-07-01

FOXP2, a forkhead box-containing transcription factor, forms homo- or hetero-dimers with FOXP family members and localizes to the nucleus, while FOXP2(R553H), which contains a mutation related to speech/language disorders, features reduced DNA binding activity and both cytoplasmic and nuclear localization. In addition to being a loss-of-function mutation, it is possible that FOXP2(R553H) also may act as a gain-of-function mutation to inhibit the functions of FOXP2 isoforms including FOXP2Ex10+ lacking forkhead domain. Foxp2(R552H) knock-in mouse pups exhibit impaired ultrasonic vocalization and poor dendritic development in Purkinje cells. However, expressions of Foxp2 isoforms in the developing Purkinje are unclear. The appearance of 'apical cytoplasmic swelling' (mitochondria-rich regions that are the source of budding processes) correlates with dendritic development of Purkinje cells. In the present study, we focused on Foxp2 isoforms localizing to the apical cytoplasmic swelling and identified two isoforms lacking forkhead domain: Foxp2Ex12+ and Foxp2Ex15. They partly localized to the membrane fraction that includes mitochondria. Foxp2Ex12+ mainly localized to the apical cytoplasmic swelling in early developing Purkinje cells at the stellate stage (P2-P4). Mitochondrial localization of Foxp2Ex12+ in Purkinje cells was confirmed by immune-electron microscopic analysis. Foxp2Ex12+ may play a role in dendritic development in Purkinje cells. © 2011 The Authors. Journal of Neurochemistry © 2011 International Society for Neurochemistry.

Differential localization of cytoplasmic myosin II isoforms A and B in avian interphase and dividing embryonic and immortalized cardiomyocytes and other cell types in vitro

NASA Technical Reports Server (NTRS)

Conrad, A. H.; Jaffredo, T.; Conrad, G. W.; Spooner, B. S. (Principal Investigator)

1995-01-01

Two principal isoforms of cytoplasmic myosin II, A and B (CMIIA and CMIIB), are present in different proportions in different tissues. Isoform-specific monoclonal and polyclonal antibodies to avian CMIIA and CMIIB reveal the cellular distributions of these isoforms in interphase and dividing embryonic avian cardiac, intestinal epithelial, spleen, and dorsal root ganglia cells in primary cell culture. Embryonic cardiomyocytes react with antibodies to CMIIB but not to CMIIA, localize CMIIB in stress-fiber-like-structures during interphase, and markedly concentrate CMIIB in networks in the cleavage furrow during cytokinesis. In contrast, cardiac fibroblasts localize both CMIIA and CMIIB in stress fibers and networks during interphase, and demonstrate slight and independently regulated concentration of CMIIA and CMIIB in networks in their cleavage furrows. V-myc-immortalized cardiomyocytes, an established cell line, have regained the ability to express CMIIA, as well as CMIIB, and localize both CMIIA and CMIIB in stress fibers and networks in interphase cells and in cleavage furrows in dividing cells. Conversely, some intestinal epithelial, spleen, and dorsal root ganglia interphase cells express only CMIIA, organized primarily in networks. Of these, intestinal epithelial cells express both CMIIA and CMIIB when they divide, whereas some dividing cells from both spleen and dorsal root ganglia express only CMIIA and concentrate it in their cleavage furrows. These results suggest that within a given tissue, different cell types express different isoforms of CMII, and that cells expressing either CMIIA or CMIIB alone, or simultaneously, can form a cleavage furrow and divide.

The Effects of CO2 Injection and Barrel Temperatures on the Physiochemical and Antioxidant Properties of Extruded Cereals

PubMed Central

Thin, Thazin; Myat, Lin; Ryu, Gi-Hyung

2016-01-01

The effects of CO2 injection and barrel temperatures on the physiochemical and antioxidant properties of extruded cereals (sorghum, barley, oats, and millet) were studied. Extrusion was carried out using a twin-screw extruder at different barrel temperatures (80, 110, and 140°C), CO2 injection (0 and 500 mL/min), screw speed of 200 rpm, and moisture content of 25%. Extrusion significantly increased the total flavonoid content (TFC) of extruded oats, and β-glucan and protein digestibility (PD) of extruded barley and oats. In contrast, there were significant reductions in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, PD of extruded sorghum and millet, as well as resistant starch (RS) of extruded sorghum and barley, and total phenolic content (TPC) of all extrudates, except extruded millet. At a barrel temperature of 140°C, TPC in extruded barley was significantly increased, and there was also an increase in DPPH and PD in extruded millet with or without CO2 injection. In contrast, at a barrel temperature of 140°C, the TPC of extruded sorghum decreased, TFC of extruded oats decreased, and at a barrel temperature of 110°C, PD of extruded sorghum without CO2 decreased. Some physical properties [expansion ratio (ER), specific length, piece density, color, and water absorption index] of the extrudates were significantly affected by the increase in barrel temperature. The CO2 injection significantly affected some physical properties (ER, specific length, piece density, water solubility index, and water absorption index), TPC, DPPH, β-glucan, and PD. In conclusion, extruded barley and millet had higher potential for making value added cereal-based foods than the other cereals. PMID:27752504

Changes in total phenol, flavonoid contents and anti-Lactobacillus activity of Callisia fragrans due to extraction solvent

NASA Astrophysics Data System (ADS)

Le, Thom; Cao, Diem Kieu; Pham, Thanh Vy; Huynh, Tan Dat; Ta, Nhat Thuy Anh; Nguyen, Ngoc Thao Linh; Nguyen, Huu Thanh; Le, Hue Huong; Bui, Anh Vo; Truong, Dieu-Hien

2018-04-01

Callisia fragrans is a wonder herb with many medicinal properties such as burn, dental diseases, cancer diseases and arthritis in folk medicine. It is noted that the phytochemical constituents and antimicrobial activity of traditional plants depend on not only the extracting method but also the solvent used for extraction. In this study, the effect of five extraction solvents (i.e., distilled water, 80% methanol, 80% ethanol, 80% ethyl acetate, and 80% chloroform) on yield, total phenolic content (TPC) and total flavonoid content (TFC) of Callisia leaves was determined. Besides, changes in anti-Lactobacillus fermentum activity of C. fragrans freeze-dried extract was also evaluated using disk-diffusion method. The recovery percentage of extractable yield of fresh leaves are ranged from 11.93% w/w for distilled water extract to 16.60% w/w for aqueous ethanol extracts. The yield of 80% aqueous methanol extract (16.27% w/w) is only slightly less than that of the ethanol extract. Significant differences were observed among TPC and TFC obtaining by 80% methanol (0.0522% and 0.0335% w/w, respectively) compared to other solvents (p < 0.05). TPC and TFC of C. fragrans extracts increase in the following order: distilled water < 80% chloroform < 80% ethyl acetate < 80% ethanol < 80% methanol. The results revealed that 80% aqueous methanol Calissia extracts has moderate inhibition (9.0 mm of inhibition zone for 1.5 mg/mL of extracts) of L. fermentum compared to standard antibacterial agent. Based on the study results, it can be concluded that the yield, TPC and TFC of C. frgrans extract varied with the extracting solvent. It also showed that Callisia extracts can prevent dental caries by inhibiting the growth of L. fermentum, towards new insights for treatment of dental caries.

GEM detector performance with innovative micro-TPC readout in high magnetic field

NASA Astrophysics Data System (ADS)

Garzia, I.; Alexeev, M.; Amoroso, A.; Baldini Ferroli, R.; Bertani, M.; Bettoni, D.; Bianchi, F.; Calcaterra, A.; Canale, N.; Capodiferro, M.; Cassariti, V.; Cerioni, S.; Chai, J. Y.; Chiozzi, S.; Cibinetto, G.; Cossio, F.; Cotta Ramusino, A.; De Mori, F.; Destefanis, M.; Dong, J.; Evangelisti, F.; Evangelisti, F.; Farinelli, R.; Fava, L.; Felici, G.; Fioravanti, E.; Gatta, M.; Greco, M.; Lavezzi, L.; Leng, C. Y.; Li, H.; Maggiora, M.; Malaguti, R.; Marcello, S.; Melchiorri, M.; Mezzadri, G.; Mignone, M.; Morello, G.; Pacetti, S.; Patteri, P.; Pellegrino, J.; Pelosi, A.; Rivetti, A.; Rolo, M. D.; Savrié, M.; Scodeggio, M.; Soldani, E.; Sosio, S.; Spataro, S.; Tskhadadze, E.; Verma, S.; Wheadon, R.; Yan, L.

2018-01-01

Gas detector development is one of the pillars of the research in fundamental physics. Since several years, a new concept of detectors, called Micro Pattern Gas Detector (MPGD), allowed to overcome several problems related to other types of commonly used detectors, like drift chamber and micro strips detectors, reducing the rate of discharges and providing better radiation tolerance. Among the most used MPGDs are the Gas Electron Multipliers (GEMs). Invented by Sauli in 1997, nowadays GEMs have become an important reality for particle detectors in high energy physics. Commonly deployed as fast timing detectors and triggers, their fast response, high rate capability and high radiation hardness make them also suitable as tracking detectors. The readout scheme is one of the most important features in tracking technology. Analog readout based on the calculation of the center of gravity technique allows to overcome the limit imposed by digital pads, whose spatial resolution is limited by the pitch dimensions. However, the presence of high external magnetic fields can distort the electronic cloud and affect the performance. The development of the micro-TPC reconstruction method brings GEM detectors into a new prospective, improving significantly the spatial resolutionin presence of high magnetic fields. This innovative technique allows to reconstruct the 3-dimensional particle position, as Time Projection Chamber, but within a drift gap of a few millimeters. In these report, the charge centroid and micro-TPC methods are described in details. We discuss the results of several test beams performed with planar chambers in magnetic field. These results are one of the first developments of micro-TPC technique for GEM detectors, which allows to reach unprecedented performance in a high magnetic field of 1 T.

Prolonged survival in patients with breast cancer and a history of brain metastases: results of a preplanned subgroup analysis from the randomized phase III BEACON trial.

PubMed

Cortés, Javier; Rugo, Hope S; Awada, Ahmad; Twelves, Chris; Perez, Edith A; Im, Seock-Ah; Gómez-Pardo, Patricia; Schwartzberg, Lee S; Diéras, Veronique; Yardley, Denise A; Potter, David A; Mailliez, Audrey; Moreno-Aspitia, Alvaro; Ahn, Jin-Seok; Zhao, Carol; Hoch, Ute; Tagliaferri, Mary; Hannah, Alison L; O'Shaughnessy, Joyce

2017-09-01

Conventional chemotherapy has limited activity in patients with breast cancer and brain metastases (BCBM). Etirinotecan pegol (EP), a novel long-acting topoisomerase-1 inhibitor, was designed using advanced polymer technology to preferentially accumulate in tumor tissue including brain metastases, providing sustained cytotoxic SN38 levels. The phase 3 BEACON trial enrolled 852 women with heavily pretreated locally recurrent or metastatic breast cancer between 2011 and 2013. BEACON compared EP with treatment of physician's choice (TPC; eribulin, vinorelbine, gemcitabine, nab-paclitaxel, paclitaxel, ixabepilone, or docetaxel) in patients previously treated with anthracycline, taxane, and capecitabine, including those with treated, stable brain metastases. The primary endpoint, overall survival (OS), was assessed in a pre-defined subgroup of BCBM patients; an exploratory post hoc analysis adjusting for the diagnosis-specific graded prognostic assessment (GPA) index was also conducted. In the trial, 67 BCBM patients were randomized (EP, n = 36; TPC, n = 31). Treatment subgroups were balanced for baseline characteristics and GPA indices. EP was associated with a significant reduction in the risk of death (HR 0.51; P < 0.01) versus TPC; median OS was 10.0 and 4.8 months, respectively. Improvement in OS was observed in both poorer and better GPA prognostic groups. Survival rates at 12 months were 44.4% for EP versus 19.4% for TPC. Consistent with the overall BEACON population, fewer patients on EP experienced grade ≥3 toxicity (50 vs. 70%). The significant improvement in survival in BCBM patients provides encouraging data for EP in this difficult-to-treat subgroup of patients. A phase three trial of EP in BCBM patients is underway (ClinicalTrials.gov NCT02915744).

Early postoperative and long-term oncological outcomes of laparoscopic treatment for patients with familial adenomatous polyposis

PubMed Central

Kim, Hye Jin; Park, Jun Seok; Park, Soo Yeun; Choi, Wohn Ho; Ryuk, Jong Pil

2012-01-01

Purpose We evaluated the short- and long-term outcomes of laparoscopic total proctocolectomy with ileal pouch-anal anastomosis (TPC/IPAA) for treatment of familial adenomatous polyposis (FAP). Also, we assessed the oncologic outcomes in FAP patients with coexisting malignancy. Methods From August 1999 to September 2010, 43 FAP patients with or without coexisting malignancy underwent TPC/IPAA by a laparoscopic-assisted or hand-assisted laparoscopic surgery. Results The median age was 33 years (range, 18 to 58 years) at the time of operation. IPAA was performed by a hand-sewn method in 21 patients (48.8%). The median operative time was 300 minutes (range, 135 to 610 minutes), which reached a plateau after 22 operations. Early postoperative complications within 30 days occurred in 7 patients (16.3%) and long-term morbidity occurred in 15 patients (34.9%) including 6 (14.0%) with desmoid tumors and 3 (7.0%) who required operative treatment. Twenty-two patients (51.2%) were diagnosed with coexisting colorectal malignancy. The median follow-up was 58.5 months (range, 7.9 to 97.8 months). There was only 1 case of local recurrence in the pelvic cavity. No cases of adenocarcinoma at the residual rectal mucosa developed. 5-year disease-free survival rate for 22 patients who had coexisting malignancy was 86.5% and 5-year overall survival rate was 92.6%. Three patients died from pulmonary or hepatic metastasis. Conclusion Laparoscopic TPC/IPAA in patients with FAP is feasible and offers favorable postoperative outcomes. It also delivered acceptable oncological outcomes in patients with coexisting malignancy. Therefore, laparoscopic TPC/IPAA may be a favorable treatment option for FAP. PMID:23166888

The evolution of aryl hydrocarbon signaling proteins: diversity of ARNT isoforms among fish species.

PubMed

Powell, W H; Hahn, M E

2000-01-01

The aryl hydrocarbon receptor nuclear translocator (ARNT) mediates aryl hydrocarbon signaling and toxicity by dimerizing with the ligand-activated aryl hydrocarbon receptor (AHR), forming a complex that binds specific DNA elements and alters transcription of target genes. Two genes encode different forms of ARNT in rodents: ARNT1, which is widely expressed, and ARNT2, which exhibits a very restricted expression pattern. In an effort to characterize aryl hydrocarbon signaling mechanisms in fishes, we previously isolated an ARNT cDNA from Fundulus heteroclitus and discovered that this species expresses ARNT2 ubiquitously. This situation differs not only from mammals, but also from rainbow trout, which expresses a divergent ARNT gene that we hypothesized was peculiar to salmonids (rtARNTa/b). In this communication, we examine the ARNT sequences of multiple fish species, including a newly isolated cDNA from scup (Stenotomus chrysops). Our phylogenetic analysis demonstrates that zebrafish ARNT, like the Fundulus protein, is an ARNT2. Contrary to expectations, the scup ARNT is closely related to the rainbow trout protein, demonstrating that the existence of this ARNT isoform predates the divergence of salmonids from the other teleosts. Thus, different species of fish express distinct and highly conserved isoforms of ARNT. The number, type, and expression pattern of ARNT proteins may contribute to interspecies differences in aryl hydrocarbon toxicity, possibly through distinct interactions with additional PAS-family proteins.

Biochemical features of the adhesion G protein-coupled receptor CD97 related to its auto-proteolysis and HeLa cell attachment activities

PubMed Central

Yang, Li-yun; Liu, Xiao-fang; Yang, Yang; Yang, Lin-lin; Liu, Kai-wen; Tang, Yu-bo; Zhang, Min; Tan, Min-jia; Cheng, Shan-mei; Xu, Ye-chun; Yang, Huai-yu; Liu, Zhi-jie; Song, Gao-jie; Huang, Wei

2017-01-01

CD97 belongs to the adhesion GPCR family characterized by a long ECD linked to the 7TM via a GPCR proteolytic site (GPS) and plays important roles in modulating cell migration and invasion. CD97 (EGF1-5) is a splicing variant of CD97 that recognizes a specific ligand chondroitin sulfate on cell membranes and the extracellular matrix. The aim of this study was to elucidate the extracellular molecular basis of the CD97 EGF1-5 isoform in protein expression, auto-proteolysis and cell adhesion, including epidermal growth factor (EGF)-like domain, GPCR autoproteolysis-inducing (GAIN) domain, as well as GPS mutagenesis and N-glycosylation. Both wild-type (WT) CD97-ECD and its truncated, GPS mutated, PNGase F-deglycosylated, and N-glycosylation site mutated forms were expressed and purified. The auto-proteolysis of the proteins was analyzed with Western blotting and SDS-PAGE. Small angle X-ray scattering (SAXS) and molecular modeling were used to determine a structural profile of the properly expressed receptor. Potential N-glycosylation sites were identified using MS and were modulated with PNGase F digestion and glyco-site mutations. A flow cytometry-based HeLa cell attachment assay was used for all aforementioned CD97 variants to elucidate the molecular basis of CD97-HeLa interactions. A unique concentration-dependent GPS auto-proteolysis was observed in CD97 EGF1-5 isoform with the highest concentration (4 mg/mL) per sample was self-cleaved much faster than the lower concentration (0.1 mg/mL), supporting an intermolecular mechanism of auto-proteolysis that is distinct to the reported intramolecular mechanism for other CD97 isoforms. N-glycosylation affected the auto-proteolysis of CD97 EGF1-5 isoform in a similar way as the other previously reported CD97 isoforms. SAXS data for WT and deglycosylated CD97ECD revealed a spatula-like shape with GAIN and EGF domains constituting the body and handle, respectively. Structural modeling indicated a potential interaction between the GAIN and EGF5 domains accounting for the absence of expression of the GAIN domain itself, although EGF5-GAIN was expressed similarly in the wild-type protein. For HeLa cell adhesion, the GAIN-truncated forms showed dramatically reduced binding affinity. The PNGase F-deglycosylated and GPS mutated forms also exhibited reduced HeLa attachment compared with WT CD97. However, neither N-glycosylation mutagenesis nor auto-proteolysis inhibition caused by N-glycosylation mutagenesis affected CD97-HeLa cell interactions. A comparison of the HeLa binding affinities of PNGase F-digested, GPS-mutated and N-glycosylation-mutated CD97 samples revealed diverse findings, suggesting that the functions of CD97 ECD were complex, and various technologies for function validation should be utilized to avoid single-approach bias when investigating N-glycosylation and auto-proteolysis of CD97. A unique mechanism of concentration-dependent auto-proteolysis of the CD97 EGF1-5 isoform was characterized, suggesting an intermolecular mechanism that is distinct from that of other previously reported CD97 isoforms. The EGF5 and GAIN domains are likely associated with each other as CD97 expression and SAXS data revealed a potential interaction between the two domains. Finally, the GAIN and EGF domains are also important for CD97-HeLa adhesion, whereas N-glycosylation of the CD97 GAIN domain and GPS auto-proteolysis are not required for HeLa cell attachment. PMID:27641734

Biochemical features of the adhesion G protein-coupled receptor CD97 related to its auto-proteolysis and HeLa cell attachment activities.

PubMed

Yang, Li-Yun; Liu, Xiao-Fang; Yang, Yang; Yang, Lin-Lin; Liu, Kai-Wen; Tang, Yu-Bo; Zhang, Min; Tan, Min-Jia; Cheng, Shan-Mei; Xu, Ye-Chun; Yang, Huai-Yu; Liu, Zhi-Jie; Song, Gao-Jie; Huang, Wei

2017-01-01

CD97 belongs to the adhesion GPCR family characterized by a long ECD linked to the 7TM via a GPCR proteolytic site (GPS) and plays important roles in modulating cell migration and invasion. CD97 (EGF1-5) is a splicing variant of CD97 that recognizes a specific ligand chondroitin sulfate on cell membranes and the extracellular matrix. The aim of this study was to elucidate the extracellular molecular basis of the CD97 EGF1-5 isoform in protein expression, auto-proteolysis and cell adhesion, including epidermal growth factor (EGF)-like domain, GPCR autoproteolysis-inducing (GAIN) domain, as well as GPS mutagenesis and N-glycosylation. Both wild-type (WT) CD97-ECD and its truncated, GPS mutated, PNGase F-deglycosylated, and N-glycosylation site mutated forms were expressed and purified. The auto-proteolysis of the proteins was analyzed with Western blotting and SDS-PAGE. Small angle X-ray scattering (SAXS) and molecular modeling were used to determine a structural profile of the properly expressed receptor. Potential N-glycosylation sites were identified using MS and were modulated with PNGase F digestion and glyco-site mutations. A flow cytometry-based HeLa cell attachment assay was used for all aforementioned CD97 variants to elucidate the molecular basis of CD97-HeLa interactions. A unique concentration-dependent GPS auto-proteolysis was observed in CD97 EGF1-5 isoform with the highest concentration (4 mg/mL) per sample was self-cleaved much faster than the lower concentration (0.1 mg/mL), supporting an intermolecular mechanism of auto-proteolysis that is distinct to the reported intramolecular mechanism for other CD97 isoforms. N-glycosylation affected the auto-proteolysis of CD97 EGF1-5 isoform in a similar way as the other previously reported CD97 isoforms. SAXS data for WT and deglycosylated CD97ECD revealed a spatula-like shape with GAIN and EGF domains constituting the body and handle, respectively. Structural modeling indicated a potential interaction between the GAIN and EGF5 domains accounting for the absence of expression of the GAIN domain itself, although EGF5-GAIN was expressed similarly in the wild-type protein. For HeLa cell adhesion, the GAIN-truncated forms showed dramatically reduced binding affinity. The PNGase F-deglycosylated and GPS mutated forms also exhibited reduced HeLa attachment compared with WT CD97. However, neither N-glycosylation mutagenesis nor auto-proteolysis inhibition caused by N-glycosylation mutagenesis affected CD97-HeLa cell interactions. A comparison of the HeLa binding affinities of PNGase F-digested, GPS-mutated and N-glycosylation-mutated CD97 samples revealed diverse findings, suggesting that the functions of CD97 ECD were complex, and various technologies for function validation should be utilized to avoid single-approach bias when investigating N-glycosylation and auto-proteolysis of CD97. A unique mechanism of concentration-dependent auto-proteolysis of the CD97 EGF1-5 isoform was characterized, suggesting an intermolecular mechanism that is distinct from that of other previously reported CD97 isoforms. The EGF5 and GAIN domains are likely associated with each other as CD97 expression and SAXS data revealed a potential interaction between the two domains. Finally, the GAIN and EGF domains are also important for CD97-HeLa adhesion, whereas N-glycosylation of the CD97 GAIN domain and GPS auto-proteolysis are not required for HeLa cell attachment.

Comparative genomic analysis reveals a novel mitochondrial isoform of human rTS protein and unusual phylogenetic distribution of the rTS gene

PubMed Central

Liang, Ping; Nair, Jayakumar R; Song, Lei; McGuire, John J; Dolnick, Bruce J

2005-01-01

Background The rTS gene (ENOSF1), first identified in Homo sapiens as a gene complementary to the thymidylate synthase (TYMS) mRNA, is known to encode two protein isoforms, rTSα and rTSβ. The rTSβ isoform appears to be an enzyme responsible for the synthesis of signaling molecules involved in the down-regulation of thymidylate synthase, but the exact cellular functions of rTS genes are largely unknown. Results Through comparative genomic sequence analysis, we predicted the existence of a novel protein isoform, rTS, which has a 27 residue longer N-terminus by virtue of utilizing an alternative start codon located upstream of the start codon in rTSβ. We observed that a similar extended N-terminus could be predicted in all rTS genes for which genomic sequences are available and the extended regions are conserved from bacteria to human. Therefore, we reasoned that the protein with the extended N-terminus might represent an ancestral form of the rTS protein. Sequence analysis strongly predicts a mitochondrial signal sequence in the extended N-terminal of human rTSγ, which is absent in rTSβ. We confirmed the existence of rTS in human mitochondria experimentally by demonstrating the presence of both rTSγ and rTSβ proteins in mitochondria isolated by subcellular fractionation. In addition, our comprehensive analysis of rTS orthologous sequences reveals an unusual phylogenetic distribution of this gene, which suggests the occurrence of one or more horizontal gene transfer events. Conclusion The presence of two rTS isoforms in mitochondria suggests that the rTS signaling pathway may be active within mitochondria. Our report also presents an example of identifying novel protein isoforms and for improving gene annotation through comparative genomic analysis. PMID:16162288

Cellular localization and changes in expression of prolactin receptor isoforms in sheep ovary throughout the estrous cycle.

PubMed

Picazo, R A; García Ruiz, J P; Santiago Moreno, J; González de Bulnes, A; Muñoz, J; Silván, G; Lorenzo, P L; Illera, J C

2004-11-01

The actions of prolactin (PRL) on target cells depend on the type of prolactin receptor (PRLr) predominantly expressed, particularly whether the long PRLr isoform is expressed. The aims of this study were to determine the cellular localization and the changes in expression of long and short PRLr isoforms in sheep ovary throughout the estrous cycle. Long and short PRLrs were localized mostly in the same ovarian cells. Maximum signal intensity, particularly for long PRLrs, was found in stromal cells surrounding primordial and primary follicles, and, for both PRLrs, in granulosa cells of preantral follicles and in luteal cells. Moderate signal intensity for PRLrs was found in theca cells of preantral to ovulatory follicles, and in granulosa cells of antral follicles up to the gonadotropin-dependent stage. Decreasing immunoreactivity to PRLrs was found in granulosa cells of gonadotropin-dependent to ovulatory follicles. For long PRLrs in particular, no signal was found in mural granulosa cells of gonadotropin-dependent follicles; for both isoforms, no signal was found in most granulosa cells of ovulatory follicles. In primordial to gonadotropin-dependent follicles, cellular localization of PRLr was similar on days 0, 10 and 15 of the cycle. Oocytes consistently showed positive immunostaining for PRLrs. Comparative RT-PCR analysis of long and short PRLr expression showed that the short isoform is evenly expressed throughout the estrous cycle, whereas the expression of the long form increases at the time of estrus and decreases at mid-luteal phase and at the onset of the follicular phase. Expression of long PRLrs was greater than that of short PRLrs on day 0 of cycle; expression of both isoforms was similar on day 10 and on day 15, long PRLrs expression was lower than that of short PRLrs. Our results indicate that in sheep ovary, the maximum responsiveness to PRL might occur during the preovulatory phase of the estrous cycle.

Charged residues distribution modulates selectivity of the open state of human isoforms of the voltage dependent anion-selective channel.

PubMed

Amodeo, Giuseppe Federico; Scorciapino, Mariano Andrea; Messina, Angela; De Pinto, Vito; Ceccarelli, Matteo

2014-01-01

Voltage Dependent Anion-selective Channels (VDACs) are pore-forming proteins located in the outer mitochondrial membrane. They are responsible for the access of ions and energetic metabolites into the inner membrane transport systems. Three VDAC isoforms exist in mammalian, but their specific role is unknown. In this work we have performed extensive (overall ∼5 µs) Molecular Dynamics (MD) simulations of the human VDAC isoforms to detect structural and conformational variations among them, possibly related to specific functional roles of these proteins. Secondary structure analysis of the N-terminal domain shows a high similarity among the three human isoforms of VDAC but with a different plasticity. In particular, the N-terminal domain of the hVDAC1 is characterized by a higher plasticity, with a ∼20% occurrence for the 'unstructured' conformation throughout the folded segment, while hVDAC2, containing a peculiar extension of 11 amino acids at the N-terminal end, presents an additional 310-helical folded portion comprising residues 10' to 3, adhering to the barrel wall. The N-terminal sequences of hVDAC isoforms are predicted to have a low flexibility, with possible consequences in the dynamics of the human VDACs. Clear differences were found between hVDAC1 and hVDAC3 against hVDAC2: a significantly modified dynamics with possible important consequence on the voltage-gating mechanism. Charge distribution inside and at the mouth of the pore is responsible for a different preferential localization of ions with opposite charge and provide a valuable rationale for hVDAC1 and hVDAC3 having a Cl-/K+ selectivity ratio of 1.8, whereas hVDAC2 of 1.4. Our conclusion is that hVDAC isoforms, despite sharing a similar scaffold, have modified working features and a biological work is now requested to give evidence to the described dissimilarities.

Na/K-ATPase regulates bovine sperm capacitation through raft- and non-raft-mediated signaling mechanisms.

PubMed

Rajamanickam, Gayathri D; Kastelic, John P; Thundathil, Jacob C

2017-11-01

Highly dynamic lipid microdomains (rafts) in the sperm plasma membrane contain several signaling proteins that regulate sperm capacitation. Na/K-ATPase isoforms (testis-specific isoform ATP1A4 and ubiquitous isoform ATP1A1) are abundant in bovine sperm plasma membrane. We previously reported that incubation of bovine sperm with ouabain, a specific Na/K-ATPase ligand, induced tyrosine phosphorylation of several sperm proteins during capacitation. The objective of this study was to investigate the roles of lipid rafts and non-rafts in Na/K-ATPase enzyme activity and signaling during bovine sperm capacitation. Content of ATP1A4 and, to a lesser extent, ATP1A1 was increased in raft and non-raft fractions of capacitated sperm, although non-raft enzyme activities of both isoforms were higher than the corresponding activities in rafts from capacitated sperm. Yet, ATP1A4 was the predominant isoform responsible for total Na/K-ATPase activity in both rafts and non-rafts. A comparative increase in phosphorylation of signaling molecules was observed in both raft (CAV1) and non-raft (EGFR and ERK1/2) membrane fractions during capacitation. Although SRC was phosphorylated in both membrane fractions, the non-raft fraction possessed more of this activated form. We also inferred, by immunoprecipitation, that ATP1A4 interacted with CAV1 and EGFR in the raft fraction, whereas interactions of ATP1A4 with SRC, EGFR, and ERK1/2 occurred in the non-raft fraction of ouabain-capacitated sperm; conversely, ATP1A1 interacted only with CAV1 in both fractions of uncapacitated and capacitated sperm. In conclusion, both raft and non-raft cohorts of Na/K-ATPase isoforms contributed to phosphorylation of signaling molecules during bovine sperm capacitation. © 2017 Wiley Periodicals, Inc.

Precise 3D Track Reconstruction Algorithm for the ICARUS T600 Liquid Argon Time Projection Chamber Detector

DOE PAGES

Antonello, M.; Baibussinov, B.; Benetti, P.; ...

2013-01-15

Liquid Argon Time Projection Chamber (LAr TPC) detectors offer charged particle imaging capability with remarkable spatial resolution. Precise event reconstruction procedures are critical in order to fully exploit the potential of this technology. In this paper we present a new, general approach to 3D reconstruction for the LAr TPC with a practical application to the track reconstruction. The efficiency of the method is evaluated on a sample of simulated tracks. We present also the application of the method to the analysis of stopping particle tracks collected during the ICARUS T600 detector operation with the CNGS neutrino beam.

A Combinatorial Geometry Computer Description of the M578 Light Recovery Vehicle

DTIC Science & Technology

1984-05-01

cannot overlap. 10 TABLE 1. GEOMETRIC SOLIDS USED IN COM-GEOM DESCRIPTIONS Symbol Solid Name RPP Rectangular Parallelepiped BOX Box RAW Right Angle...20R «OX 209 PCC 210 RCC 211 TRC 212 RHX "»13 RCC 214 RCC 2T5 TRC 216 BOX ?17 PrC ?"»R R^C SOLID PARAMETERS REMARKS 74.0303 3694.444...821720 «OX 221 RCC 22’ PC* 223 TPC 224 30V 225 "CC 2?6 PCC 227 TRC 22* BOX 220 RCC 230 »CC 231 TRC ?3’ TPC 233 TRC 234 RCC SOLID

DOE Office of Scientific and Technical Information (OSTI.GOV)

Acciarri, R.; Adams, C.; An, R.

We present several studies of convolutional neural networks applied to data coming from the MicroBooNE detector, a liquid argon time projection chamber (LArTPC). The algorithms studied include the classification of single particle images, the localization of single particle and neutrino interactions in an image, and the detection of a simulated neutrino event overlaid with cosmic ray backgrounds taken from real detector data. These studies demonstrate the potential of convolutional neural networks for particle identification or event detection on simulated neutrino interactions. We also address technical issues that arise when applying this technique to data from a large LArTPC at ormore » near ground level.« less

The CERES/NA45 radial drift Time Projection Chamber

NASA Astrophysics Data System (ADS)

Adamová, D.; Agakichiev, G.; Antończyk, D.; Appelshäuser, H.; Belaga, V.; Bielčíková, J.; Braun-Munzinger, P.; Campagnolo, R.; Cherlin, A.; Damjanović, S.; Dietel, T.; Dietrich, L.; Drees, A.; Dubitzky, W.; Esumi, S. I.; Filimonov, K.; Fraenkel, Z.; Garabatos, C.; Glässel, P.; Hering, G.; Holeczek, J.; Kushpil, V.; Marín, A.; Milošević, J.; Milov, A.; Miśkowiec, D.; Musa, L.; Panebrattsev, Y.; Pechenova, O.; Petráček, V.; Pfeiffer, A.; Rak, J.; Ravinovich, I.; Richter, M.; Sako, H.; Schäfer, E.; Schmitz, W.; Schukraft, J.; Seipp, W.; Sharma, A.; Shimansky, S.; Stachel, J.; Šumbera, M.; Tilsner, H.; Tserruya, I.; Wessels, J. P.; Wienold, T.; Windelband, B.; Wurm, J. P.; Xie, W.; Yurevich, S.; Yurevich, V.

2008-08-01

The design, calibration, and performance of the first radial drift Time Projection Chamber (TPC) are presented. The TPC was built and installed at the CERES/NA45 experiment at the CERN SPS in the late nineties, with the objective to improve the momentum resolution of the spectrometer. The upgraded experiment took data twice, in 1999 and in 2000. After a detailed study of residual distortions a spatial resolution of 340 μm in the azimuthal and 640 μm in the radial direction was achieved, corresponding to a momentum resolution of Δp/p=√{(1%·p/GeV)2+(2%)2}.

Design of the cryogenic systems for the Near and Far LAr-TPC detectors of the Short-Baseline Neutrino program (SBN) at Fermilab

DOE Office of Scientific and Technical Information (OSTI.GOV)

Geynisman, M.; Bremer, J.; Chalifour, M.

The Short-Baseline Neutrino (SBN) physics program at Fermilab and Neutrino Platform (NP) at CERN are part of the international Neutrino Program leading to the development of Long-Baseline Neutrino Facility/Deep Underground Neutrino Experiment (LBNF/DUNE) science project. The SBN program consisting of three Liquid Argon Time Projection Chamber (LAr-TPC) detectors positioned along the Booster Neutrino Beam (BNB) at Fermilab includes an existing detector known as MicroBooNE (170-ton LAr-TPC) plus two new experiments known as SBN’s Near Detector (SBND, ~260 tons) and SBN’s Far Detector (SBN-FD, ~760 tons). All three detectors have distinctly different design of their cryostats thus defining specific requirements formore » the cryogenic systems. Fermilab has already built two new facilities to house SBND and SBN-FD detectors. The cryogenic systems for these detectors are in various stages of design and construction with CERN and Fermilab being responsible for delivery of specific sub-systems. This contribution presents specific design requirements and typical implementation solutions for each sub-system of the SBND and SBN-FD cryogenic systems.« less

Exploring 0.1–10 eV axions with a new helioscope concept

DOE Office of Scientific and Technical Information (OSTI.GOV)

Galán, J.; Dafni, T.; Iguaz, F.J., E-mail: javier.galan.lacarra@cern.ch, E-mail: Theopisti.Dafni@cern.ch, E-mail: iguaz@unizar.es

2015-12-01

We explore the possibility to develop a new axion helioscope type, sensitive to the higher axion mass region favored by axion models. We propose to use a low background large volume TPC immersed in an intense magnetic field. Contrary to traditional tracking helioscopes, this detection technique takes advantage of the capability to directly detect the photons converted on the buffer gas which defines the axion mass sensitivity region, and does not require pointing the magnet to the Sun. The operation flexibility of a TPC to be used with different gas mixtures (He, Ne, Xe, etc.) and pressures (from 10 mbarmore » to 10 bar) will allow to enhance sensitivity for axion masses from few meV to several eV. We present different helioscope data taking scenarios, considering detection efficiency and axion absorption probability, and show the sensitivities reachable with this technique to be few × 10{sup −11} GeV{sup −1} for a 5 T, m{sup 3} scale TPC. We show that a few years program taking data with such setup would allow to probe the KSVZ axion model for axion masses above 0∼> 10 meV.« less

Microbiological evaluation of South Australian rock lobster meat.

PubMed

Yap, A S

1977-12-01

Samples of frozen precooked rock lobster meat from five South Australian fish-processing plants situated in the West Coast and south-east regions were tested over a period of six months during the 1974/5 lobster fishing season. The most probable number (MPN) of E. coli and coliforms, Staphylococcus aureus and Salmonella, as well as total plate count (TPC) were determined in 480 samples. Monthly geometric mean TPC ranged from 1600/g to 25,000/g. The highest geometric mean of the MPN of coliforms and E. coli were 4.9/g and 1.8/g respectively. The highest geometric mean number of staphylococci was 18.6/g. Salmonella was not detected in the 480 units tested. Only 0.4% of the samples had TPC exceeding 100,000/g. Coliforms and E. coli were not present in 76.1% and 92.7% respectively of the samples tested. Staphylococcus aureus was not detected in 67.7% of the samples. The numbers of organisms in 82% of the samples fall within the microbiological standards proposed by the National Health and Medical Research Council of Australia for frozen precooked foods. The results of this study demonstrate the microbial quality of precooked lobster meat attainable when good manufacturing practices are used.

Research on the effect of culture time on the kombucha tea beverage's antiradical capacity and sensory value.

PubMed

Gramza-Michałowska, Anna; Kulczyński, Bartosz; Xindi, Yuan; Gumienna, Małgorzata

2016-01-01

Recent consumption trends shows high consumer acceptability and growing medicinal interest in the biological value of kombucha tea. This tea is a sweetened tea leaf brew fermented with a layer containing mainly acetic acid bacteria, yeast and lactic acid bacteria. The main antioxidants in tea leaves are polyphenols, the consumption of which is proven to be beneficial for human health, e.g. protecting from reactive oxygen species (ROS). The aim of the present research was to evaluate antiradical activity, total polyphenol content (TPC) and sensory value of kombucha tea brews. In the present study, Kombucha tea beverages were analyzed for TPC content, DPPH radical scavenging method and sensory value. The highest TPC content and DPPH radical scavenging capacity values were evaluated in yellow tea samples, both unfermented and kombucha, which did not differ within the storage time. The results of sensory evaluations of kombucha tea brews depend on the tea leaf variety used for preparing the drink. Research indicates that the fermentation process of tea brews with kombucha microbiota does not affect significantly its polyphenol content and antiradical capacity, and retains its components' biological activity.

Measurement of the normalized U 238 ( n , f ) / U 235 ( n , f ) cross section ratio from threshold to 30 MeV with the NIFFTE fission Time Projection Chamber

DOE PAGES

Casperson, R. J.; Asner, D. M.; Baker, J.; ...

2018-03-23

We present that the normalized 238U(n,f)/ 235U(n,f) cross section ratio has been measured using the NIFFTE fission Time Projection Chamber (fissionTPC) from the reaction threshold to 30 MeV . The fissionTPC is a two-volume MICROMEGAS time projection chamber that allows for full three-dimensional reconstruction of fission-fragment ionization profiles from neutron-induced fission. The measurement was performed at the Los Alamos Neutron Science Center, where the neutron energy is determined from neutron time of-flight. The 238U(n,f)/ 235U(n,f) ratio reported here is the first cross section measurement made with the fissionTPC, and will provide new experimental data for evaluation of the 238U(n,f) crossmore » section, an important standard used in neutron-flux measurements. Use of a development target in this work prevented the determination of an absolute normalization, to be addressed in future measurements. Instead, the measured cross section ratio has been normalized to ENDF/B-VIII.β5 at 14.5 MeV.« less

Catechin Composition and Antioxidant Activity of Black Teas in Relation to Brewing Time.

PubMed

Koch, Wojciech; Kukula-Koch, Wirginia; Głowniak, Kazimierz

2017-11-01

Black tea infusions are one of the most popular beverages across the world. Their extract composition depends on several factors, brewing time being one of the most important determinants. The aim of the present study was to determine the catechin composition of different black tea infusions using a validated LC electrospray ionization time-of-flight MS method. Additionally, total phenolic content (TPC) and antioxidant activity of infusions were evaluated using Folin-Ciocalteu reagent and stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). An optimized LC-MS method enabled the precise identification of the studied catechins [epicatechin (EC), EC gallate (ECG), epigallocatechin (EGC), and epigallocatechin-3-gallate (EGCG)] and gallic acid (GA). The major catechin in all investigated teas was EGC (25.6 mg/100 cm3 after 4 min of brewing). EC was present at the lowest concentration in all extracts. TPC and antiradical scavenging activity were in a good agreement with catechins and GA content. In general, the longer the brewing time, the higher the concentration of catechin, TPC, and antioxidant activity values. However, it should be noted that after 2 min brewing, most phenolics had already been extracted, and extract composition did not significantly change at a prolonged extraction time.

Josephson Photodetectors via Temperature-to-Phase Conversion

NASA Astrophysics Data System (ADS)

Virtanen, P.; Ronzani, A.; Giazotto, F.

2018-05-01

We theoretically investigate the temperature-to-phase conversion (TPC) process occurring in dc superconducting quantum interferometers based on superconductor-normal-metal-superconductor (S -N -S ) mesoscopic Josephson junctions. In particular, we predict the temperature-driven rearrangement of the phase gradients in the interferometer under the fixed constraints of fluxoid quantization and supercurrent conservation. This mechanism allows sizeable phase variations across the junctions for suitable structure parameters and temperatures. We show that the TPC can be a basis for sensitive single-photon sensors or bolometers. We propose a radiation detector realizable with conventional materials and state-of-the-art nanofabrication techniques. Integrated with a superconducting quantum-interference proximity transistor as a readout setup, an aluminum-based TPC calorimeter can provide a large signal-to-noise ratio >100 in the 10-GHz-10-THz frequency range and a resolving power larger than 1 02 below 50 mK for terahertz photons. In the bolometric operation, electrical noise equivalent power of approximately 10-22 W /√{Hz } is predicted at 50 mK. This device can be attractive as a cryogenic single-photon sensor operating in the giga- and terahertz regime with applications in dark-matter searches.

Design of the cryogenic systems for the Near and Far LAr-TPC detectors of the Short-Baseline Neutrino program (SBN) at Fermilab

NASA Astrophysics Data System (ADS)

Geynisman, M.; Bremer, J.; Chalifour, M.; Delaney, M.; Dinnon, M.; Doubnik, R.; Hentschel, S.; Kim, M. J.; Montanari, C.; Montanari, D.; Nichols, T.; Norris, B.; Sarychev, M.; Schwartz, F.; Tillman, J.; Zuckerbrot, M.

2017-12-01

The Short-Baseline Neutrino (SBN) physics program at Fermilab and Neutrino Platform (NP) at CERN are part of the international Neutrino Program leading to the development of Long-Baseline Neutrino Facility/Deep Underground Neutrino Experiment (LBNF/DUNE) science project. The SBN program consisting of three Liquid Argon Time Projection Chamber (LAr-TPC) detectors positioned along the Booster Neutrino Beam (BNB) at Fermilab includes an existing detector known as MicroBooNE (170-ton LAr-TPC) plus two new experiments known as SBN’s Near Detector (SBND, ∼260 tons) and SBN’s Far Detector (SBN-FD, ∼760 tons). All three detectors have distinctly different design of their cryostats thus defining specific requirements for the cryogenic systems. Fermilab has already built two new facilities to house SBND and SBN-FD detectors. The cryogenic systems for these detectors are in various stages of design and construction with CERN and Fermilab being responsible for delivery of specific sub-systems. This contribution presents specific design requirements and typical implementation solutions for each sub-system of the SBND and SBN-FD cryogenic systems.

Effect of drying methods on total antioxidant capacity of bitter gourd (momordica charantia) fruit

NASA Astrophysics Data System (ADS)

Tan, Ee Shian; Abdullah, Aminah; Maskat, Mohammad Yusof

2013-11-01

The effect of thermal and non-thermal drying methods on hydrophilic and lipophilic antioxidant capacities of bitter gourd fruit was investigated in this study. The bitter gourd fruits were dried by following methods: (i) oven drying 40°C, (ii) oven drying 50°C, (iii) oven drying 60°C, (iv) microwave drying (medium low power), (v) microwave drying (medium power) and (vi) freeze drying. Pure acetone and hexane were used to extract the hydrophilic and lipophilic antioxidant compounds from dried bitter gourd fruits. Freeze dried extracts reported to have highest values in DPPH scavenging activity (hydrophilic and lipophilic fractions), FRAP (lipophilic fraction) and TPC (hydrophilic and lipophilic fraction). Thermal drying slightly increased the values of DPPH scavenging activity, FRAP and TPC assays for hydrophilic extracts. Results concluded bitter gourd fruit is a good source of natural antioxidants and its total antioxidant quality was most preserved by freeze drying. Additionally, the higher value reported in DPPH scavenging activity, FRAP and TPC assays for lipophilic extracts than the hydrophilic extracts suggested that the lipophilic antioxidant compounds of bitter gourd fruit might possess stronger antioxidant power than its counterpart.

Measurement of the normalized U 238 ( n , f ) / U 235 ( n , f ) cross section ratio from threshold to 30 MeV with the NIFFTE fission Time Projection Chamber

DOE Office of Scientific and Technical Information (OSTI.GOV)

Casperson, R. J.; Asner, D. M.; Baker, J.

We present that the normalized 238U(n,f)/ 235U(n,f) cross section ratio has been measured using the NIFFTE fission Time Projection Chamber (fissionTPC) from the reaction threshold to 30 MeV . The fissionTPC is a two-volume MICROMEGAS time projection chamber that allows for full three-dimensional reconstruction of fission-fragment ionization profiles from neutron-induced fission. The measurement was performed at the Los Alamos Neutron Science Center, where the neutron energy is determined from neutron time of-flight. The 238U(n,f)/ 235U(n,f) ratio reported here is the first cross section measurement made with the fissionTPC, and will provide new experimental data for evaluation of the 238U(n,f) crossmore » section, an important standard used in neutron-flux measurements. Use of a development target in this work prevented the determination of an absolute normalization, to be addressed in future measurements. Instead, the measured cross section ratio has been normalized to ENDF/B-VIII.β5 at 14.5 MeV.« less

Antioxidant and cytoprotective activities of Piper betle, Areca catechu, Uncaria gambir and betel quid with and without calcium hydroxide.

PubMed

Sazwi, Nordin Nur; Nalina, Thurairajah; Abdul Rahim, Zubaidah Haji

2013-12-11

Betel quid chewing is a popular habit in Southeast Asia. It is believed that chewing betel quid could reduce stress, strengthen teeth and maintain oral hygiene. The aim of this study was to investigate the antioxidant and cytoprotective activities of each of the ingredients of betel quid and compared with betel quid itself (with and without calcium hydroxide). The correlation of their cytoprotective and antioxidant activities with phenolic content was also determined. Five samples (betel leaf, areca nut, gambir, betel quid and betel quid containing calcium hydroxide) were extracted in deionized distilled water for 12 hours at 37°C. Antioxidant activities were evaluated for radical scavenging activity using DPPH assay, ferric reducing activity using FRAP assay and lipid peroxidation inhibition activity using FTC assay. Total phenolic content (TPC) was determined using Folin-Ciocalteu procedure. Phenolic composition was analyzed using LC-MS/MS. Cytoprotective activity towards human gingival fibroblast cells was examined using MTT assay. Among the ingredients of betel quid, gambir demonstrated the highest antioxidant (DPPH - IC50 = 6.4 ± 0.8 μg/mL, FRAP - 5717.8 ± 537.6 μmol Fe(II)/mg), total phenolic content (TPC - 1142.5 ± 106.8 μg TAE/mg) and cytoprotective (100.1 ± 4.6%) activities. Betel quid when compared with betel quid containing calcium hydroxide has higher antioxidant (DPPH - IC50 =59.4 ± 4.4 μg/mL, FRAP - 1022.2 ± 235.7 μmol Fe(II)/mg), total phenolic content (TPC - 140.0 ± 22.3 μg TAE/mg), and cytoprotective (113.5 ± 15.9%) activities. However, all of the five samples showed good lipid peroxidation inhibition compared to vitamin E. LC-MS/MS analysis revealed the presence of quinic acid as the major compound of gambir and betel quid. A positive correlation was observed between TPC and radical scavenging (r = 0.972), reducing power (r = 0.981) and cytoprotective activity (r = 0.682). The betel quid has higher TPC, and antioxidant and cytoprotective activities than betel quid with calcium hydroxide. The quinic acid in betel quid may play an important role in the oral health protection.

Evaluation of phenolic composition, antioxidant, anti-inflammatory and anticancer activities of Polygonatum verticillatum (L.).

PubMed

Kumar Singh, Sandeep; Patra, Arjun

2018-04-18

Polygonatum verticillatum (L.) All. (Ruscaceae), one of the Ashtawarga plants, is widely used for treatment of various ailments. The present study was undertaken to determine the phenolic composition, antioxidant, anti-inflammatory and anticancer activities of several extracts (petroleum ether, dichloromethane, chloroform, ethanol, and aqueous) from the rhizomes of the plant. Coarsely powdered dry rhizome was successively extracted with different solvents of increasing polarity (petroleum ether, dichloromethane, chloroform, ethanol and water). The phenolic compositions, in terms of total phenolic content (TPC), total flavonoid content (TFC) and total condensed tannin content (TTC), were evaluated with the Folin-Ciocalteu assay, aluminum chloride colorimetric assay and vanillin spectrophotometric assay, respectively. Total antioxidant capacity, 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assays were used to assess the antioxidant potential of each extract. A protein denaturation model and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were used to evaluate in vitro anti-inflammatory and anticancer activities, respectively. Gas chromatography-mass spectrometry (GC/MS) analysis was carried out to demonstrate various phytoconstituents in each extract. Correlation studies were also performed between phenolic composition (TPC, TFC and TTC) and different biological activities. Ethanol extract showed maximum TPC (0.126 mg/g, gallic acid equivalent in dry sample), TFC (0.094 mg/g, rutin equivalent in dry sample) and TTC (29.32 mg/g, catechin equivalent in dry sample), as well as antioxidant and anti-inflammatory properties. Chloroform extract exhibited the strongest cytotoxicity against the human breast cancer cell line, MCF-7. GC/MS analysis revealed the presence of 90 different phytoconstituents among the extracts. Antioxidant and anti-inflammatory activities had a positive correlation with TPC, TFC and TTC. However, the anticancer activity showed a negative correlation with TPC, TFC and TTC. From the present study, it can be concluded that P. verticillatum possessed remarkable antioxidant, anti-inflammatory, and anticancer activities, which could be due to different secondary metabolites of the plant. Phenolic compounds are likely responsible for antioxidant and anti-inflammatory activities. However, flavonoids and other compounds might contribute to the anticancer potential of the plant. Copyright © 2018 Shanghai Changhai Hospital. Published by Elsevier B.V. All rights reserved.

Neutron Fluence and Energy Reconstruction with the LNE-IRSN/MIMAC Recoil Detector MicroTPC at 27 keV

DOE Office of Scientific and Technical Information (OSTI.GOV)

Maire, D.; Lebreton, L.; Querre, Ph.

2015-07-01

The French Institute for Radiation protection and Nuclear Safety (IRSN), designated by the French Metrology Institute (LNE) for neutron metrology, is developing a time projection chamber using a Micromegas anode: microTPC. This work is carried out in collaboration with the Laboratory of Subatomic Physics and Cosmology (LPSC). The aim is to characterize the energy distribution of neutron fluence in the energy range 8 keV - 5 MeV with a primary procedure. The time projection chambers are gaseous detectors able to measure charged particles energy and to reconstruct their track if a pixelated anode is used. In our case, the gasmore » is used as a (n, p) converter in order to detect neutrons down to few keV. Coming from elastic collisions with neutrons, recoil protons lose a part of their kinetic energy by ionizing the gas. The ionization electrons are drifted toward a pixelated anode (2D projection), read at 50 MHz by a self-triggered electronic system to obtain the third track dimension. The neutron energy is reconstructed event by event thanks to proton scattering angle and proton energy measurements. The scattering angle is deduced from the 3D track. The proton energy is obtained by charge collection measurements, knowing the ionization quenching factor (i.e. the part of proton kinetic energy lost by ionizing the gas). The fluence is calculated thanks to the detected events number and the simulation of the detector response. The μTPC is a new reliable detector able to measure energy distribution of the neutron fluence without unfolding procedure or prior neutron calibration contrary to usual gaseous counters. The microTPC is still being developed and measurements have been carried out at the AMANDE facility, with neutrons energies going from 8 keV to 565 keV. After the context and the μ-TPC working principle presentation, measurements of the neutron energy and fluence at 27 keV and 144 keV are shown and compared to the complete detector response simulation. This work shows the first direct reconstruction of neutron energy and fluence, simultaneously, at 27.2 keV in a continuous irradiation mode. (authors)« less

Neutron fluence and energy reconstruction with the IRSN recoil detector μ-TPC at 27 keV, 144 keV and 565 keV

DOE Office of Scientific and Technical Information (OSTI.GOV)

Maire, D.; Lebreton, L.; Richer, J.P.

2015-07-01

The French Institute for Radioprotection and Nuclear Safety (IRSN), associated to the French Metrology Institute (LNE), is developing a time projection chamber using a Micromegas anode: μ-TPC. This work is carried out in collaboration with the Laboratory of Subatomic Physics and Cosmology (LPSC). The aim is to characterize with a primary procedure the energy distribution of neutron fluence in the energy range 8 keV - 1 MeV. The time projection chambers are gaseous detectors, which are able to measure charged particles energy and to reconstruct their track if a pixelated anode is used. In our case, the gas is usedmore » as a (n, p) converter in order to detect neutrons down to few keV. Coming from elastic collisions with neutrons, recoil protons lose a part of their kinetic energy by ionizing the gas. The ionization electrons are drifted toward a pixelated anode (2D projection), read at 50 MHz by a self-triggered electronic system to obtain the third track dimension. The neutron energy is reconstructed event by event thanks to proton scattering angle and proton energy measurements. The scattering angle is deduced from the 3D track. The proton energy is obtained by charge collection measurements, knowing the ionization quenching factor (i.e. the part of proton kinetic energy lost by ionizing the gas). The fluence is calculated thanks to the detected events number and the simulated detector response. The μ-TPC is a new reliable detector which enables to measure energy distribution of the neutron fluence without deconvolution or neutron calibration contrary to usual gaseous counters. The μ-TPC is still being developed and measurements have been carried out at the AMANDE facility, with neutrons energies going from 8 keV to 565 keV. After the context and the μ-TPC working principle presentation, measurements of the neutron energy and fluence at 27.2 keV, 144 keV and 565 keV are shown and compared to the complete detector simulation. This work shows the first direct reconstruction of neutron energy and fluence, simultaneously, at 27.2 keV in a continuous irradiation mode. (authors)« less

NAADP Activates Two-Pore Channels on T Cell Cytolytic Granules to Stimulate Exocytosis and Killing

PubMed Central

Davis, Lianne C.; Morgan, Anthony J.; Chen, Ji-Li; Snead, Charlotte M.; Bloor-Young, Duncan; Shenderov, Eugene; Stanton-Humphreys, Megan N.; Conway, Stuart J.; Churchill, Grant C.; Parrington, John; Cerundolo, Vincenzo; Galione, Antony

2012-01-01

Summary A cytotoxic T lymphocyte (CTL) kills an infected or tumorigenic cell by Ca2+-dependent exocytosis of cytolytic granules at the immunological synapse formed between the two cells. Although inositol 1,4,5-trisphosphate (IP3)-mediated Ca2+ release from the endoplasmic reticulum activates the store-operated Ca2+-influx pathway that is necessary for exocytosis, it is not a sufficient stimulus [1–4]. Here we identify the Ca2+-mobilizing messenger nicotinic acid adenine dinucleotide phosphate (NAADP) and its recently identified molecular target, two-pore channels (TPCs) [5–7], as being important for T cell receptor signaling in CTLs. We demonstrate that cytolytic granules are not only reservoirs of cytolytic proteins but are also the acidic Ca2+ stores mobilized by NAADP via TPC channels on the granules themselves, so that TPCs migrate to the immunological synapse upon CTL activation. Moreover, NAADP activates TPCs to drive exocytosis in a way that is not mimicked by global Ca2+ signals induced by IP3 or ionomycin, suggesting that critical, local Ca2+ nanodomains around TPCs stimulate granule exocytosis. Hence, by virtue of the NAADP/TPC pathway, cytolytic granules generate Ca2+ signals that lead to their own exocytosis and to cell killing. This study highlights a selective role for NAADP in stimulating exocytosis crucial for immune cell function and may impact on stimulus-secretion coupling in wider cellular contexts. PMID:23177477

Changes in chemical composition and antioxidative properties of rye ginger cakes during their shelf-life.

PubMed

Zieliński, Henryk; del Castillo, Maria Dolores; Przygodzka, Małgorzata; Ciesarova, Zuzana; Kukurova, Kristina; Zielińska, Danuta

2012-12-15

Changes in chemical composition and antioxidative properties of rye ginger cakes during their shelf-life were investigated in this study. In particular, the changes in antioxidants content, antioxidative and reducing capacity, and Maillard reaction development in rye ginger cakes after long-term storage were addressed. Ginger cakes produced according to the traditional and current recipe were stored for 5 years at room temperature in a dark place. The total phenolic compounds (TPC), inositol hexaphosphate (IP6), reduced (GSH) and oxidised glutathione (GSSG) contents, antioxidant and reducing capacity and Maillard reaction products (MRPs) were determined in ginger cakes after storage and then compared to those measured after baking. After long-term storage a decrease in TPC and IP6 contents in cakes was noted. In contrast, an increase in antioxidative and reducing capacity of stored cakes was observed. Long-term storage induced formation of furosine, advanced and final Maillard reaction products and caused changes in both reduced and oxidised forms of glutathione. After long-term storage the modest changes in furosine, FAST index and browning in ginger cake formulated with dark rye flour may suggest that this product is the healthiest among others. Therefore, traditional rye ginger cakes can be considered as an example of a healthy food that is also relatively stable during long term storage as noted by the small chemical changes observed in its composition. Copyright © 2012 Elsevier Ltd. All rights reserved.

Online Calibration of the TPC Drift Time in the ALICE High Level Trigger

NASA Astrophysics Data System (ADS)

Rohr, David; Krzewicki, Mikolaj; Zampolli, Chiara; Wiechula, Jens; Gorbunov, Sergey; Chauvin, Alex; Vorobyev, Ivan; Weber, Steffen; Schweda, Kai; Lindenstruth, Volker

2017-06-01

A Large Ion Collider Experiment (ALICE) is one of the four major experiments at the Large Hadron Collider (LHC) at CERN. The high level trigger (HLT) is a compute cluster, which reconstructs collisions as recorded by the ALICE detector in real-time. It employs a custom online data-transport framework to distribute data and workload among the compute nodes. ALICE employs subdetectors that are sensitive to environmental conditions such as pressure and temperature, e.g., the time projection chamber (TPC). A precise reconstruction of particle trajectories requires calibration of these detectors. Performing calibration in real time in the HLT improves the online reconstructions and renders certain offline calibration steps obsolete speeding up offline physics analysis. For LHC Run 3, starting in 2020 when data reduction will rely on reconstructed data, online calibration becomes a necessity. Reconstructed particle trajectories build the basis for the calibration making a fast online-tracking mandatory. The main detectors used for this purpose are the TPC and Inner Tracking System. Reconstructing the trajectories in the TPC is the most compute-intense step. We present several improvements to the ALICE HLT developed to facilitate online calibration. The main new development for online calibration is a wrapper that can run ALICE offline analysis and calibration tasks inside the HLT. In addition, we have added asynchronous processing capabilities to support long-running calibration tasks in the HLT framework, which runs event-synchronously otherwise. In order to improve the resiliency, an isolated process performs the asynchronous operations such that even a fatal error does not disturb data taking. We have complemented the original loop-free HLT chain with ZeroMQ data-transfer components. The ZeroMQ components facilitate a feedback loop that inserts the calibration result created at the end of the chain back into tracking components at the beginning of the chain, after a short delay. All these new features are implemented in a general way, such that they have use-cases aside from online calibration. In order to gather sufficient statistics for the calibration, the asynchronous calibration component must process enough events per time interval. Since the calibration is valid only for a certain time period, the delay until the feedback loop provides updated calibration data must not be too long. A first full-scale test of the online calibration functionality was performed during 2015 heavy-ion run under real conditions. Since then, online calibration is enabled and benchmarked in 2016 proton-proton data taking. We present a timing analysis of this first online-calibration test, which concludes that the HLT is capable of online TPC drift time calibration fast enough to calibrate the tracking via the feedback loop. We compare the calibration results with the offline calibration and present a comparison of the residuals of the TPC cluster coordinates with respect to offline reconstruction.

IsoPlot: a database for comparison of mRNA isoforms in fruit fly and mosquitoes

PubMed Central

Ng, I-Man; Tsai, Shang-Chi

2017-01-01

Abstract Alternative splicing (AS), a mechanism by which different forms of mature messenger RNAs (mRNAs) are generated from the same gene, widely occurs in the metazoan genomes. Knowledge about isoform variants and abundance is crucial for understanding the functional context in the molecular diversity of the species. With increasing transcriptome data of model and non-model species, a database for visualization and comparison of AS events with up-to-date information is needed for further research. IsoPlot is a publicly available database with visualization tools for exploration of AS events, including three major species of mosquitoes, Aedes aegypti, Anopheles gambiae, and Culex quinquefasciatus, and fruit fly Drosophila melanogaster, the model insect species. IsoPlot includes not only 88,663 annotated transcripts but also 17,037 newly predicted transcripts from massive transcriptome data at different developmental stages of mosquitoes. The web interface enables users to explore the patterns and abundance of isoforms in different experimental conditions as well as cross-species sequence comparison of orthologous transcripts. IsoPlot provides a platform for researchers to access comprehensive information about AS events in mosquitoes and fruit fly. Our database is available on the web via an interactive user interface with an intuitive graphical design, which is applicable for the comparison of complex isoforms within or between species. Database URL: http://isoplot.iis.sinica.edu.tw/ PMID:29220459

The non-canonical functions of the heme oxygenases

PubMed Central

Tibullo, Daniele; Forte, Stefano; Zappalà, Agata; Volti, Giovanni Li

2016-01-01

Heme oxygenase (HO) isoforms catalyze the conversion of heme to carbon monoxide (CO) and biliverdin with a concurrent release of iron, which can drive the synthesis of ferritin for iron sequestration. Most of the studies so far were directed at evaluating the protective effect of these enzymes because of their ability to generate antioxidant and antiapoptotic molecules such as CO and bilirubin. Recent evidences are suggesting that HO may possess other important physiological functions, which are not related to its enzymatic activity and for which we would like to introduce for the first time the term “non canonical functions”. Recent evidence suggest that both HO isoforms may form protein-protein interactions (i.e. cytochrome P450, adiponectin, CD91) thus serving as chaperone-like protein. In addition, truncated HO-1 isoform was localized in the nuclear compartment under certain experimental conditions (i.e. excitotoxicity, hypoxia) regulating the activity of important nuclear transcription factors (i.e. Nrf2) and DNA repair. In the present review, we discuss three potential signaling mechanisms that we refer to as the non-canonical functions of the HO isoforms: protein-protein interaction, intracellular compartmentalization, and extracellular secretion. The aim of the present review is to describe each of this mechanism and all the aspects warranting additional studies in order to unravel all the functions of the HO system. PMID:27626166

Role of Leptin Deficiency, Inefficiency, and Leptin Receptors in Obesity.

PubMed

Wasim, Muhammad; Awan, Fazli Rabbi; Najam, Syeda Sadia; Khan, Abdul Rehman; Khan, Haq Nawaz

2016-10-01

Leptin protein consists of 167 amino acids, which is mainly secreted from the white adipose tissue. This protein acts on the hypothalamic regions of the brain which control eating behavior, thus playing a significant role in maintaining body's metabolism. Leptin receptors belong to glycoprotein 130 (gp130) family of cytokine receptors and exist in six isoforms (LEPR a-f), and all the isoforms are encoded by LEPR gene; out of these isoforms, the LEPR-b receptor is the 'longest form,' and in most of the cases, mutations in this isoform cause severe obesity. Also, mutations in the leptin gene (LEP) or its receptors gene can lead to obesity. Some biochemical pathways affect the bioactivity of leptin and/or its receptors. To date, eleven pathogenic mutations have been reported in the LEP which are p.L72S, p.N103K, p.R105W, p.H118L, p.S141C, p.W121X c.104_106delTCA, c.135del3bp, c.398delG, c.481_482delCT, and c.163C>T. Different mutations in the LEPR have also been reported as c.2396-1 G>T, c.1675 G>A, p.P316T, etc. In some studies, where leptin was deficient, leptin replacement therapy has shown positive impact by preventing weight gain and obesity.

Splice form variant and amino acid changes in MDR49 confers DDT resistance in transgenic Drosophila

PubMed Central

Seong, Keon Mook; Sun, Weilin; Clark, John M.; Pittendrigh, Barry R.

2016-01-01

The ATP-binding cassette (ABC) transporters represent a superfamily of proteins that have important physiological roles in both prokaryotes and eukaryotes. In insects, ABC transporters have previously been implicated in insecticide resistance. The 91-R strain of Drosophila melanogaster has been intensely selected with DDT over six decades. A recent selective sweeps analysis of 91-R implicated the potential role of MDR49, an ABC transporter, in DDT resistance, however, to date the details of how MDR49 may play a role in resistance have not been elucidated. In this study, we investigated the impact of structural changes and an alternative splicing event in MDR49 on DDT-resistance in 91-R, as compared to the DDT susceptible strain 91-C. We observed three amino acid differences in MDR49 when 91-R was compared with 91-C, and only one isoform (MDR49B) was implicated in DDT resistance. A transgenic Drosophila strain containing the 91-R-MDR49B isoform had a significantly higher LD50 value as compared to the 91-C-MDR49B isoform at the early time points (6 h to 12 h) during DDT exposure. Our data support the hypothesis that the MDR49B isoform, with three amino acid mutations, plays a role in the early aspects of DDT resistance in 91-R. PMID:27003579

Splice form variant and amino acid changes in MDR49 confers DDT resistance in transgenic Drosophila.

PubMed

Seong, Keon Mook; Sun, Weilin; Clark, John M; Pittendrigh, Barry R

2016-03-22

The ATP-binding cassette (ABC) transporters represent a superfamily of proteins that have important physiological roles in both prokaryotes and eukaryotes. In insects, ABC transporters have previously been implicated in insecticide resistance. The 91-R strain of Drosophila melanogaster has been intensely selected with DDT over six decades. A recent selective sweeps analysis of 91-R implicated the potential role of MDR49, an ABC transporter, in DDT resistance, however, to date the details of how MDR49 may play a role in resistance have not been elucidated. In this study, we investigated the impact of structural changes and an alternative splicing event in MDR49 on DDT-resistance in 91-R, as compared to the DDT susceptible strain 91-C. We observed three amino acid differences in MDR49 when 91-R was compared with 91-C, and only one isoform (MDR49B) was implicated in DDT resistance. A transgenic Drosophila strain containing the 91-R-MDR49B isoform had a significantly higher LD50 value as compared to the 91-C-MDR49B isoform at the early time points (6 h to 12 h) during DDT exposure. Our data support the hypothesis that the MDR49B isoform, with three amino acid mutations, plays a role in the early aspects of DDT resistance in 91-R.

Method for the Simultaneous Quantitation of Apolipoprotein E Isoforms using Tandem Mass Spectrometry

PubMed Central

Wildsmith, Kristin R.; Han, Bomie; Bateman, Randall J.

2009-01-01

Using Apolipoprotein E (ApoE) as a model protein, we developed a protein isoform analysis method utilizing Stable Isotope Labeling Tandem Mass Spectrometry (SILT MS). ApoE isoforms are quantitated using the intensities of the b and y ions of the 13C-labeled tryptic isoform-specific peptides versus unlabeled tryptic isoform-specific peptides. The ApoE protein isoform analysis using SILT allows for the simultaneous detection and relative quantitation of different ApoE isoforms from the same sample. This method provides a less biased assessment of ApoE isoforms compared to antibody-dependent methods, and may lead to a better understanding of the biological differences between isoforms. PMID:19653990

Altered STAT4 Isoform Expression in Patients with Inflammatory Bowel Disease.

PubMed

Jabeen, Rukhsana; Miller, Lucy; Yao, Weiguo; Gupta, Sandeep; Steiner, Steven; Kaplan, Mark H

2015-10-01

Crohn's disease (CD) and ulcerative colitis (UC) are the major forms of inflammatory bowel disease, and pathogenesis involves a complex interplay among genetic, environmental, and immunological factors. We evaluated isoform expression of the IL-12-activated transcription factor STAT4 in children with CD and UC. We collected biopsy samples from both patients newly diagnosed with CD and with UC. We further collected blood samples from patients newly diagnosed with CD and with UC as well as from patients who had a flare-up after being in clinical remission, and we examined the ratios of STAT4β/STAT4α mRNA. In addition to STAT4 isoforms, we measured the expression of the cytokines TNFα, IFNγ, granulocyte macrophage-colony stimulating factor, and IL-17 using polymerase chain reaction of biopsy samples and multiplex analysis of patient serum samples. Ratios of STAT4β/STAT4α were increased in specific gastrointestinal tract segments in both patients with CD and those with UC that correlate with the location and severity of inflammation. In contrast, we did not observe changes in STAT4β/STAT4α ratios in biopsy specimens from patients with eosinophilic esophagitis. We also observed increased STAT4β/STAT4α ratios in the peripheral blood mononuclear cells of patients with UC and those with CD, compared with healthy controls. Ratios were normalized after patients were treated with steroids. Collectively, these data indicate that STAT4 isoforms could be an important noninvasive biomarker in the diagnosis and treatment of inflammatory bowel disease and that expression of these isoforms might provide further insight into the pathogenesis of IBD.

Transgenic mouse α- and β-cardiac myosins containing the R403Q mutation show isoform-dependent transient kinetic differences.

PubMed

Lowey, Susan; Bretton, Vera; Gulick, James; Robbins, Jeffrey; Trybus, Kathleen M

2013-05-24

Familial hypertrophic cardiomyopathy (FHC) is a major cause of sudden cardiac death in young athletes. The discovery in 1990 that a point mutation at residue 403 (R403Q) in the β-myosin heavy chain (MHC) caused a severe form of FHC was the first of many demonstrations linking FHC to mutations in muscle proteins. A mouse model for FHC has been widely used to study the mechanochemical properties of mutated cardiac myosin, but mouse hearts express α-MHC, whereas the ventricles of larger mammals express predominantly β-MHC. To address the role of the isoform backbone on function, we generated a transgenic mouse in which the endogenous α-MHC was partially replaced with transgenically encoded β-MHC or α-MHC. A His6 tag was cloned at the N terminus, along with R403Q, to facilitate isolation of myosin subfragment 1 (S1). Stopped flow kinetics were used to measure the equilibrium constants and rates of nucleotide binding and release for the mouse S1 isoforms bound to actin. For the wild-type isoforms, we found that the affinity of MgADP for α-S1 (100 μM) is ~ 4-fold weaker than for β-S1 (25 μM). Correspondingly, the MgADP release rate for α-S1 (350 s(-1)) is ~3-fold greater than for β-S1 (120 s(-1)). Introducing the R403Q mutation caused only a minor reduction in kinetics for β-S1, but R403Q in α-S1 caused the ADP release rate to increase by 20% (430 s(-1)). These transient kinetic studies on mouse cardiac myosins provide strong evidence that the functional impact of an FHC mutation on myosin depends on the isoform backbone.

Cell Elasticity Is Regulated by the Tropomyosin Isoform Composition of the Actin Cytoskeleton

PubMed Central

Jalilian, Iman; Heu, Celine; Cheng, Hong; Freittag, Hannah; Desouza, Melissa; Stehn, Justine R.; Bryce, Nicole S.; Whan, Renee M.; Hardeman, Edna C.

2015-01-01

The actin cytoskeleton is the primary polymer system within cells responsible for regulating cellular stiffness. While various actin binding proteins regulate the organization and dynamics of the actin cytoskeleton, the proteins responsible for regulating the mechanical properties of cells are still not fully understood. In the present study, we have addressed the significance of the actin associated protein, tropomyosin (Tpm), in influencing the mechanical properties of cells. Tpms belong to a multi-gene family that form a co-polymer with actin filaments and differentially regulate actin filament stability, function and organization. Tpm isoform expression is highly regulated and together with the ability to sort to specific intracellular sites, result in the generation of distinct Tpm isoform-containing actin filament populations. Nanomechanical measurements conducted with an Atomic Force Microscope using indentation in Peak Force Tapping in indentation/ramping mode, demonstrated that Tpm impacts on cell stiffness and the observed effect occurred in a Tpm isoform-specific manner. Quantitative analysis of the cellular filamentous actin (F-actin) pool conducted both biochemically and with the use of a linear detection algorithm to evaluate actin structures revealed that an altered F-actin pool does not absolutely predict changes in cell stiffness. Inhibition of non-muscle myosin II revealed that intracellular tension generated by myosin II is required for the observed increase in cell stiffness. Lastly, we show that the observed increase in cell stiffness is partially recapitulated in vivo as detected in epididymal fat pads isolated from a Tpm3.1 transgenic mouse line. Together these data are consistent with a role for Tpm in regulating cell stiffness via the generation of specific populations of Tpm isoform-containing actin filaments. PMID:25978408

Molecular modelling indicates that the pathological conformations of prion proteins might be beta-helical.

PubMed Central

Downing, D T; Lazo, N D

1999-01-01

Creutzfeldt-Jakob disease, kuru, scrapie and bovine spongiform encephalopathy are diseases of the mammalian central nervous system that involve the conversion of a cellular protein into an insoluble extracellular isoform. Spectroscopic studies have shown that the precursor protein contains mainly alpha-helical and random-coil conformations, whereas the prion isoform is largely in the beta conformation. The pathogenic prion is resistant to denaturation and protease digestion and can promote the conversion of the precursor protein to the pathogenic form. These properties have yet to be explained in terms of the structural conformations of the proteins. In the present study, molecular modelling showed that prion proteins could adopt the beta-helical conformation, which has been established for a number of fibrous proteins and has been suggested previously as the basis of amyloid fibrils. The beta-helical conformation provides explanations for the biophysical and biochemical stability of prions, their ability to form templates for the transmission of pathological conformation, and the existence of phenotypical strains of the prion diseases. PMID:10510313

aPKCλ/ι and aPKCζ Contribute to Podocyte Differentiation and Glomerular Maturation

PubMed Central

Hartleben, Björn; Widmeier, Eugen; Suhm, Martina; Worthmann, Kirstin; Schell, Christoph; Helmstädter, Martin; Wiech, Thorsten; Walz, Gerd; Leitges, Michael; Schiffer, Mario

2013-01-01

Precise positioning of the highly complex interdigitating podocyte foot processes is critical to form the normal glomerular filtration barrier, but the molecular programs driving this process are unknown. The protein atypical protein kinase C (aPKC)—a component of the Par complex, which localizes to tight junctions and interacts with slit diaphragm proteins—may play a role. Here, we found that the combined deletion of the aPKCλ/ι and aPKCζ isoforms in podocytes associated with incorrectly positioned centrosomes and Golgi apparatus and mislocalized molecules of the slit diaphragm. Furthermore, aPKC-deficient podocytes failed to form the normal network of foot processes, leading to defective glomerular maturation with incomplete capillary formation and mesangiolysis. Our results suggest that aPKC isoforms orchestrate the formation of the podocyte processes essential for normal glomerular development and kidney function. Defective aPKC signaling results in a dramatically simplified glomerular architecture, causing severe proteinuria and perinatal death. PMID:23334392

Isoform-specific antibodies reveal distinct subcellular localizations of C9orf72 in amyotrophic lateral sclerosis.

PubMed

Xiao, Shangxi; MacNair, Laura; McGoldrick, Philip; McKeever, Paul M; McLean, Jesse R; Zhang, Ming; Keith, Julia; Zinman, Lorne; Rogaeva, Ekaterina; Robertson, Janice

2015-10-01

A noncoding hexanucleotide repeat expansion in C9orf72 is the most common cause of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). It has been reported that the repeat expansion causes a downregulation of C9orf72 transcripts, suggesting that haploinsufficiency may contribute to disease pathogenesis. Two protein isoforms are generated from three alternatively spliced transcripts of C9orf72; a long form (C9-L) and a short form (C9-S), and their function(s) are largely unknown owing to lack of specific antibodies. To investigate C9orf72 protein properties, we developed novel antibodies that recognize either C9-L or C9-S. Multiple techniques, including Western blot, immunohistochemistry, and coimmunoprecipitation, were used to determine the expression levels and subcellular localizations of C9-L and C9-S. Investigation of expression of C9-L and C9-S demonstrated distinct biochemical profiles, region-specific changes, and distinct subcellular localizations in ALS tissues. In particular, C9-L antibody exhibited a diffuse cytoplasmic staining in neurons and labeled large speckles in cerebellar Purkinje cells. In contrast, C9-S antibody gave very specific labeling of the nuclear membrane in healthy neurons, with apparent relocalization to the plasma membrane of diseased motor neurons in ALS. Coimmunoprecipitation experiments revealed an interaction of the C9-isoforms with both Importin β1 and Ran-GTPase, components of the nuclear pore complex. Using these antibodies, we have shown that C9orf72 may be involved in nucleocytoplasmic shuttling and this may have relevance to pathophysiology of ALS/FTLD. Our antibodies have provided improved detection of C9orf72 protein isoforms, which will help elucidate its physiological function and role in ALS/FTLD. © 2015 The Authors Annals of Neurology published by Wiley Periodicals, Inc. on behalf of American Neurological Association.

Isoform switching facilitates period control in the Neurospora crassa circadian clock.

PubMed

Akman, Ozgur E; Locke, James C W; Tang, Sanyi; Carré, Isabelle; Millar, Andrew J; Rand, David A

2008-01-01

A striking and defining feature of circadian clocks is the small variation in period over a physiological range of temperatures. This is referred to as temperature compensation, although recent work has suggested that the variation observed is a specific, adaptive control of period. Moreover, given that many biological rate constants have a Q(10) of around 2, it is remarkable that such clocks remain rhythmic under significant temperature changes. We introduce a new mathematical model for the Neurospora crassa circadian network incorporating experimental work showing that temperature alters the balance of translation between a short and long form of the FREQUENCY (FRQ) protein. This is used to discuss period control and functionality for the Neurospora system. The model reproduces a broad range of key experimental data on temperature dependence and rhythmicity, both in wild-type and mutant strains. We present a simple mechanism utilising the presence of the FRQ isoforms (isoform switching) by which period control could have evolved, and argue that this regulatory structure may also increase the temperature range where the clock is robustly rhythmic.

Neutron diffraction of acetazolamide-bound human carbonic anhydrase II reveals atomic details of drug binding

PubMed Central

Fisher, S. Zoë; Aggarwal, Mayank; Kovalevsky, Andrey Y.; Silverman, David N.; McKenna, Robert

2012-01-01

Carbonic anhydrases (CAs) catalyze the hydration of CO2 forming HCO3− and a proton, an important reaction for many physiological processes including respiration, fluid secretion, and pH regulation. As such, CA isoforms are prominent clinical targets for treating various diseases. The clinically used acetazolamide (AZM) is a sulfonamide that binds with high affinity to human CA isoform II (HCA II). There are several X-ray structures available of AZM bound to various CA isoforms, but these complexes do not show the charged state of AZM, or hydrogen (H) atom positions of the protein and solvent. Neutron diffraction is a useful technique for directly observing H atoms and the mapping of H-bonding networks that can greatly contribute to rational drug design. To this end the neutron structure of H/D exchanged HCA II crystals in complex with AZM was determined. The structure reveals the molecular details of AZM binding and the charged state of the bound drug. This represents the first determined neutron structure of a clinically used drug bound to its target. PMID:22928733

Alternative Splice Forms Influence Functions of Whirlin in Mechanosensory Hair Cell Stereocilia.

PubMed

Ebrahim, Seham; Ingham, Neil J; Lewis, Morag A; Rogers, Michael J C; Cui, Runjia; Kachar, Bechara; Pass, Johanna C; Steel, Karen P

2016-05-03

WHRN (DFNB31) mutations cause diverse hearing disorders: profound deafness (DFNB31) or variable hearing loss in Usher syndrome type II. The known role of WHRN in stereocilia elongation does not explain these different pathophysiologies. Using spontaneous and targeted Whrn mutants, we show that the major long (WHRN-L) and short (WHRN-S) isoforms of WHRN have distinct localizations within stereocilia and also across hair cell types. Lack of both isoforms causes abnormally short stereocilia and profound deafness and vestibular dysfunction. WHRN-S expression, however, is sufficient to maintain stereocilia bundle morphology and function in a subset of hair cells, resulting in some auditory response and no overt vestibular dysfunction. WHRN-S interacts with EPS8, and both are required at stereocilia tips for normal length regulation. WHRN-L localizes midway along the shorter stereocilia, at the level of inter-stereociliary links. We propose that differential isoform expression underlies the variable auditory and vestibular phenotypes associated with WHRN mutations. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Platelets lacking PIP5KIγ have normal integrin activation but impaired cytoskeletal-membrane integrity and adhesion

PubMed Central

Wang, Yanfeng; Zhao, Liang; Suzuki, Aae; Lian, Lurong; Min, Sang H.; Wang, Ziqian; Litvinov, Rustem I.; Stalker, Timothy J.; Yago, Tadayuki; Klopocki, Arkadiusz G.; Schmidtke, David W.; Yin, Helen; Choi, John K.; McEver, Rodger P.; Weisel, John W.; Hartwig, John H.; Abrams, Charles S.

2013-01-01

Three isoforms of phosphatidylinositol-4-phosphate 5-kinase (PIP5KIα, PIP5KIβ, and PIP5KIγ) can each catalyze the final step in the synthesis of phosphatidylinositol-4,5-bisphosphate (PIP2), which in turn can be either converted to second messengers or bind directly to and thereby regulate proteins such as talin. A widely quoted model speculates that only p90, a longer splice form of platelet-specific PIP5KIγ, but not the shorter p87 PIP5KIγ, regulates the ligand-binding activity of integrins via talin. However, when we used mice genetically engineered to lack only p90 PIP5KIγ, we found that p90 PIP5KIγ is not critical for integrin activation or platelet adhesion on collagen. However, p90 PIP5KIγ-null platelets do have impaired anchoring of their integrins to the underlying cytoskeleton. Platelets lacking both the p90 and p87 PIP5KIγ isoforms had normal integrin activation and actin dynamics, but impaired anchoring of their integrins to the cytoskeleton. Most importantly, they formed weak shear-resistant adhesions ex vivo and unstable vascular occlusions in vivo. Together, our studies demonstrate that, although PIP5KIγ is essential for normal platelet function, individual isoforms of PIP5KIγ fulfill unique roles for the integrin-dependent integrity of the membrane cytoskeleton and for the stabilization of platelet adhesion. PMID:23372168

Convolutional neural networks applied to neutrino events in a liquid argon time projection chamber

NASA Astrophysics Data System (ADS)

Acciarri, R.; Adams, C.; An, R.; Asaadi, J.; Auger, M.; Bagby, L.; Baller, B.; Barr, G.; Bass, M.; Bay, F.; Bishai, M.; Blake, A.; Bolton, T.; Bugel, L.; Camilleri, L.; Caratelli, D.; Carls, B.; Castillo Fernandez, R.; Cavanna, F.; Chen, H.; Church, E.; Cianci, D.; Collin, G. H.; Conrad, J. M.; Convery, M.; Crespo-Anadón, J. I.; Del Tutto, M.; Devitt, D.; Dytman, S.; Eberly, B.; Ereditato, A.; Escudero Sanchez, L.; Esquivel, J.; Fleming, B. T.; Foreman, W.; Furmanski, A. P.; Garvey, G. T.; Genty, V.; Goeldi, D.; Gollapinni, S.; Graf, N.; Gramellini, E.; Greenlee, H.; Grosso, R.; Guenette, R.; Hackenburg, A.; Hamilton, P.; Hen, O.; Hewes, J.; Hill, C.; Ho, J.; Horton-Smith, G.; James, C.; de Vries, J. Jan; Jen, C.-M.; Jiang, L.; Johnson, R. A.; Jones, B. J. P.; Joshi, J.; Jostlein, H.; Kaleko, D.; Karagiorgi, G.; Ketchum, W.; Kirby, B.; Kirby, M.; Kobilarcik, T.; Kreslo, I.; Laube, A.; Li, Y.; Lister, A.; Littlejohn, B. R.; Lockwitz, S.; Lorca, D.; Louis, W. C.; Luethi, M.; Lundberg, B.; Luo, X.; Marchionni, A.; Mariani, C.; Marshall, J.; Martinez Caicedo, D. A.; Meddage, V.; Miceli, T.; Mills, G. B.; Moon, J.; Mooney, M.; Moore, C. D.; Mousseau, J.; Murrells, R.; Naples, D.; Nienaber, P.; Nowak, J.; Palamara, O.; Paolone, V.; Papavassiliou, V.; Pate, S. F.; Pavlovic, Z.; Porzio, D.; Pulliam, G.; Qian, X.; Raaf, J. L.; Rafique, A.; Rochester, L.; von Rohr, C. Rudolf; Russell, B.; Schmitz, D. W.; Schukraft, A.; Seligman, W.; Shaevitz, M. H.; Sinclair, J.; Snider, E. L.; Soderberg, M.; Söldner-Rembold, S.; Soleti, S. R.; Spentzouris, P.; Spitz, J.; St. John, J.; Strauss, T.; Szelc, A. M.; Tagg, N.; Terao, K.; Thomson, M.; Toups, M.; Tsai, Y.-T.; Tufanli, S.; Usher, T.; Van de Water, R. G.; Viren, B.; Weber, M.; Weston, J.; Wickremasinghe, D. A.; Wolbers, S.; Wongjirad, T.; Woodruff, K.; Yang, T.; Zeller, G. P.; Zennamo, J.; Zhang, C.

2017-03-01

We present several studies of convolutional neural networks applied to data coming from the MicroBooNE detector, a liquid argon time projection chamber (LArTPC). The algorithms studied include the classification of single particle images, the localization of single particle and neutrino interactions in an image, and the detection of a simulated neutrino event overlaid with cosmic ray backgrounds taken from real detector data. These studies demonstrate the potential of convolutional neural networks for particle identification or event detection on simulated neutrino interactions. We also address technical issues that arise when applying this technique to data from a large LArTPC at or near ground level.

Novel particle tracking algorithm based on the Random Sample Consensus Model for the Active Target Time Projection Chamber (AT-TPC)

NASA Astrophysics Data System (ADS)

Ayyad, Yassid; Mittig, Wolfgang; Bazin, Daniel; Beceiro-Novo, Saul; Cortesi, Marco

2018-02-01

The three-dimensional reconstruction of particle tracks in a time projection chamber is a challenging task that requires advanced classification and fitting algorithms. In this work, we have developed and implemented a novel algorithm based on the Random Sample Consensus Model (RANSAC). The RANSAC is used to classify tracks including pile-up, to remove uncorrelated noise hits, as well as to reconstruct the vertex of the reaction. The algorithm, developed within the Active Target Time Projection Chamber (AT-TPC) framework, was tested and validated by analyzing the 4He+4He reaction. Results, performance and quality of the proposed algorithm are presented and discussed in detail.

Large Area Coverage of a TPC Endcap with GridPix Detectors

NASA Astrophysics Data System (ADS)

Kaminski, Jochen

2018-02-01

The Large Prototype TPC at DESY, Hamburg, was built by the LCTPC collaboration as a testbed for new readout technologies of Time Projection Chambers. Up to seven modules of about 400 cm2 each can be placed in the endcap. Three of these modules were equipped with a total of 160 GridPix detectors. This is a combination of a highly pixelated readout ASIC and a Micromegas built on top. GridPix detectors have a very high efficiency of detecting primary electrons, which leads to excellent spatial and energy resolutions. For the first time a large number of GridPix detectors has been operated and long segments of tracks have been recorded with excellent precision.

Operation of a LAr-TPC equipped with a multilayer LEM charge readout

NASA Astrophysics Data System (ADS)

Baibussinov, B.; Centro, S.; Farnese, C.; Fava, A.; Gibin, D.; Guglielmi, A.; Meng, G.; Pietropaolo, F.; Varanini, F.; Ventura, S.; Zatrimaylov, K.

2018-03-01

A novel detector for ionization signals in a single phase LAr-TPC has been experimented in the ICARINO test facility at the INFN Laboratories in Legnaro. It is based on the adoption of a multilayer Large Electron Multiplier (LEM) replacing the traditional anodic wire arrays. Cosmic muon tracks were detected allowing the measurement of energy deposition and a first determination of the signal to noise ratio. The analysis of the recorded events demonstrated the 3D reconstruction capability of this device for ionizing events in liquid Argon. The collected fraction of ionization charge is close to about 90%, with signal to noise ratio similar to that measured with more traditional wire chambers.

MicroBooNE and its Cross Section Measurement

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tsai, Yun-Tse

2017-05-22

MicroBooNE (the Micro Booster Neutrino Experiment) is a short-baseline neutrino experiment based on the technology of a liquid-argon time-projection chamber (LArTPC), and has recently completed its first year of data-taking in the Fermilab Booster Neutrino Beam. It aims to address the anomalous excess of events with an electromagnetic final state in MiniBooNE, to measure neutrino-argon interaction cross sections, and to provide relevant R\\&D for the future LArTPC experiments, such as DUNE. In these proceedings, we present the first reconstructed energy spectrum of Michel electrons from cosmic muon decays, the first kinematic distributions of the candidate muon tracks frommore » $$\

Systematically Differentiating Functions for Alternatively Spliced Isoforms through Integrating RNA-seq Data

PubMed Central

Menon, Rajasree; Wen, Yuchen; Omenn, Gilbert S.; Kretzler, Matthias; Guan, Yuanfang

2013-01-01

Integrating large-scale functional genomic data has significantly accelerated our understanding of gene functions. However, no algorithm has been developed to differentiate functions for isoforms of the same gene using high-throughput genomic data. This is because standard supervised learning requires ‘ground-truth’ functional annotations, which are lacking at the isoform level. To address this challenge, we developed a generic framework that interrogates public RNA-seq data at the transcript level to differentiate functions for alternatively spliced isoforms. For a specific function, our algorithm identifies the ‘responsible’ isoform(s) of a gene and generates classifying models at the isoform level instead of at the gene level. Through cross-validation, we demonstrated that our algorithm is effective in assigning functions to genes, especially the ones with multiple isoforms, and robust to gene expression levels and removal of homologous gene pairs. We identified genes in the mouse whose isoforms are predicted to have disparate functionalities and experimentally validated the ‘responsible’ isoforms using data from mammary tissue. With protein structure modeling and experimental evidence, we further validated the predicted isoform functional differences for the genes Cdkn2a and Anxa6. Our generic framework is the first to predict and differentiate functions for alternatively spliced isoforms, instead of genes, using genomic data. It is extendable to any base machine learner and other species with alternatively spliced isoforms, and shifts the current gene-centered function prediction to isoform-level predictions. PMID:24244129

Two-pore channels function in calcium regulation in sea star oocytes and embryos

PubMed Central

Ramos, Isabela; Reich, Adrian; Wessel, Gary M.

2014-01-01

Egg activation at fertilization is an excellent process for studying calcium regulation. Nicotinic acid adenine dinucleotide-phosphate (NAADP), a potent calcium messenger, is able to trigger calcium release, likely through two-pore channels (TPCs). Concomitantly, a family of ectocellular enzymes, the ADP-ribosyl cyclases (ARCs), has emerged as being able to change their enzymatic mode from one of nucleotide cyclization in formation of cADPR to a base-exchange reaction in the generation of NAADP. Using sea star oocytes we gain insights into the functions of endogenously expressed TPCs and ARCs in the context of the global calcium signals at fertilization. Three TPCs and one ARC were found in the sea star (Patiria miniata) that were localized in the cortex of the oocytes and eggs. PmTPCs were localized in specialized secretory organelles called cortical granules, and PmARCs accumulated in a different, unknown, set of vesicles, closely apposed to the cortical granules in the egg cortex. Using morpholino knockdown of PmTPCs and PmARC in the oocytes, we found that both calcium regulators are essential for early embryo development, and that knockdown of PmTPCs leads to aberrant construction of the fertilization envelope at fertilization and changes in cortical granule pH. The calcium signals at fertilization are not significantly altered when individual PmTPCs are silenced, but the timing and shape of the cortical flash and calcium wave are slightly changed when the expression of all three PmTPCs is perturbed concomitantly, suggesting a cooperative activity among TPC isoforms in eliciting calcium signals that may influence localized physiological activities. PMID:25377554

Phospholipase C-β in immune cells.

PubMed

Kawakami, Toshiaki; Xiao, Wenbin

2013-09-01

Great progress has recently been made in structural and functional research of phospholipase C (PLC)-β. We now understand how PLC-β isoforms (β1-β4) are activated by GTP-bound Gαq downstream of G protein-coupled receptors. Numerous studies indicate that PLC-βs participate in the differentiation and activation of immune cells that control both the innate and adaptive immune systems. The PLC-β3 isoform also interplays with tyrosine kinase-based signaling pathways, to inhibit Stat5 activation by recruiting the protein-tyrosine phosphatase SHP-1, with which PLC-β3 and Stat5 form a multi-molecular signaling platform, named SPS complex. The SPS complex has important regulatory roles in tumorigenesis and immune cell activation. Copyright © 2013 Elsevier Ltd. All rights reserved.

Phospholipase C-β in Immune Cells

PubMed Central

Kawakami, Toshiaki; Xiao, Wenbin

2013-01-01

Great progress has recently been made in structural and functional research of phospholipase C (PLC)-β. We now understand how PLC-β isoforms (β1-β4) are activated by GTP-bound Gαq downstream of G protein-coupled receptors. Numerous studies indicate that PLC-βs participate in the differentiation and activation of immune cells that control both the innate and adaptive immune systems. The PLC-β3 isoform also interplays with tyrosine kinase-based signaling pathways, to inhibit Stat5 activation by recruiting the protein-tyrosine phosphatase SHP-1, with which PLC-β3 and Stat5 form a multi-molecular signaling platform, named SPS complex. The SPS complex has important regulatory roles in tumorigenesis and immune cell activation. PMID:23981313

Merlin Isoforms 1 and 2 Both Act as Tumour Suppressors and Are Required for Optimal Sperm Maturation

PubMed Central

Zoch, Ansgar; Mayerl, Steffen; Schulz, Alexander; Greither, Thomas; Frappart, Lucien; Rübsam, Juliane; Heuer, Heike; Giovannini, Marco; Morrison, Helen

2015-01-01

The tumour suppressor Merlin, encoded by the gene NF2, is frequently mutated in the autosomal dominant disorder neurofibromatosis type II, characterised primarily by the development of schwannoma and other glial cell tumours. However, NF2 is expressed in virtually all analysed human and rodent organs, and its deletion in mice causes early embryonic lethality. Additionally, NF2 encodes for two major isoforms of Merlin of unknown functionality. Specifically, the tumour suppressor potential of isoform 2 remains controversial. In this study, we used Nf2 isoform-specific knockout mouse models to analyse the function of each isoform during development and organ homeostasis. We found that both isoforms carry full tumour suppressor functionality and can completely compensate the loss of the other isoform during development and in most adult organs. Surprisingly, we discovered that spermatogenesis is strictly dependent on the presence of both isoforms. While the testis primarily expresses isoform 1, we noticed an enrichment of isoform 2 in spermatogonial stem cells. Deletion of either isoform was found to cause decreased sperm quality as observed by maturation defects and head/midpiece abnormalities. These defects led to impaired sperm functionality as assessed by decreased sperm capacitation. Thus, we describe spermatogenesis as a new Nf2-dependent process. Additionally, we provide for the first time in vivo evidence for equal tumour suppressor potentials of Merlin isoform 1 and isoform 2. PMID:26258444

NAADP and the two-pore channel protein 1 participate in the acrosome reaction in mammalian spermatozoa

PubMed Central

Arndt, Lilli; Castonguay, Jan; Arlt, Elisabeth; Meyer, Dorke; Hassan, Sami; Borth, Heike; Zierler, Susanna; Wennemuth, Gunther; Breit, Andreas; Biel, Martin; Wahl-Schott, Christian; Gudermann, Thomas; Klugbauer, Norbert; Boekhoff, Ingrid

2014-01-01

The functional relationship between the formation of hundreds of fusion pores during the acrosome reaction in spermatozoa and the mobilization of calcium from the acrosome has been determined only partially. Hence, the second messenger NAADP, promoting efflux of calcium from lysosome-like compartments and one of its potential molecular targets, the two-pore channel 1 (TPC1), were analyzed for its involvement in triggering the acrosome reaction using a TPCN1 gene–deficient mouse strain. The present study documents that TPC1 and NAADP-binding sites showed a colocalization at the acrosomal region and that treatment of spermatozoa with NAADP resulted in a loss of the acrosomal vesicle that showed typical properties described for TPCs: Registered responses were not detectable for its chemical analogue NADP and were blocked by the NAADP antagonist trans-Ned-19. In addition, two narrow bell-shaped dose-response curves were identified with maxima in either the nanomolar or low micromolar NAADP concentration range, where TPC1 was found to be responsible for activating the low affinity pathway. Our finding that two convergent NAADP-dependent pathways are operative in driving acrosomal exocytosis supports the concept that both NAADP-gated cascades match local NAADP concentrations with the efflux of acrosomal calcium, thereby ensuring complete fusion of the large acrosomal vesicle. PMID:24451262

Principal Component Analysis of Stimulatory Effect of Synbiotic Combination of Indigenous Probiotic and Inulin on Antioxidant Activity of Soymilk.

PubMed

Choudhary, Shagun; Singh, Manisha; Sharma, Deepak; Attri, Sampan; Sharma, Kavita; Goel, Gunjan

2018-06-02

The present study aimed to screen the indigenous probiotic cultures for their effect on total phenolic contents (TPC) and associated antioxidant activities in synbiotic fermented soymilk during storage. Among 16 cultures, subtractive screening was conducted based on different tests such as acidification rate and proliferation of lactic acid bacteria (LAB) on supplementation of inulin (0-20 mM) and fructooligosaccharide (0-0.45 mM). Lactobacillus paracasei CD4 was selected as potential strain after principal component analysis (PCA) of different strains with prebiotic substrates at different concentrations. The strain was used for production of synbiotic soymilk product containing 10 mM inulin. The storage study was conducted at 4 °C for 21 days. During storage, the pH, titratable acidity, TPC, antioxidant activities, and viable cell counts (VCC) were determined. The fermentation of soymilk supplemented with 10 mM inulin did not alter the VCC; however, a decrease in pH and TPC and an increase in acidity and antioxidant activity were observed (p < 0.05). In conclusion, the synbiotic supplementation of inulin in soymilk enhanced the viability of Lactobacillus paracasei CD4 and antioxidant activity during storage under refrigeration conditions.

Influence of conventional and ultrasonic-assisted extraction on phenolic contents, betacyanin contents, and antioxidant capacity of red dragon fruit (Hylocereus polyrhizus).

PubMed

Ramli, Nurul Shazini; Ismail, Patimah; Rahmat, Asmah

2014-01-01

The aim of this study was to examine the effects of extraction methods on antioxidant capacities of red dragon fruit peel and flesh. Antioxidant capacities were measured using ethylenebenzothiozoline-6-sulfonic acid (ABTS) radical cation assay and ferric reducing antioxidant power assay (FRAP). Total phenolic content (TPC) was determined using Folin-Ciocalteu reagent while quantitative determination of total flavonoid content (TFC) was conducted using aluminium trichloride colorimetric method. Betacyanin content (BC) was measured by spectrophotometer. Red dragon fruit was extracted using conventional (CV) and ultrasonic-assisted extraction (UE) technique to determine the most efficient way of extracting its antioxidant components. Results indicated that UE increased TFC, reduced the extraction yield, BC, and TPC, but exhibited the strongest scavenging activity for the peel of red dragon fruit. In contrast, UE reduced BC, TFC, and scavenging activity but increased the yield for the flesh. Nonetheless, UE slightly increases TPC in flesh. Scavenging activity and reducing power were highly correlated with phenolic and flavonoid compounds. Conversely, the scavenging activity and reducing power were weakly correlated with betacyanin content. This work gives scientific evidences for the consideration of the type of extraction techniques for the peel and flesh of red dragon fruit in applied research and food industry.

Synthetic hardware performance analysis in virtualized cloud environment for healthcare organization.

PubMed

Tan, Chee-Heng; Teh, Ying-Wah

2013-08-01

The main obstacles in mass adoption of cloud computing for database operations in healthcare organization are the data security and privacy issues. In this paper, it is shown that IT services particularly in hardware performance evaluation in virtual machine can be accomplished effectively without IT personnel gaining access to actual data for diagnostic and remediation purposes. The proposed mechanisms utilized the hypothetical data from TPC-H benchmark, to achieve 2 objectives. First, the underlying hardware performance and consistency is monitored via a control system, which is constructed using TPC-H queries. Second, the mechanism to construct stress-testing scenario is envisaged in the host, using a single or combination of TPC-H queries, so that the resource threshold point can be verified, if the virtual machine is still capable of serving critical transactions at this constraining juncture. This threshold point uses server run queue size as input parameter, and it serves 2 purposes: It provides the boundary threshold to the control system, so that periodic learning of the synthetic data sets for performance evaluation does not reach the host's constraint level. Secondly, when the host undergoes hardware change, stress-testing scenarios are simulated in the host by loading up to this resource threshold level, for subsequent response time verification from real and critical transactions.

Influence of Conventional and Ultrasonic-Assisted Extraction on Phenolic Contents, Betacyanin Contents, and Antioxidant Capacity of Red Dragon Fruit (Hylocereus polyrhizus)

PubMed Central

Ramli, Nurul Shazini; Ismail, Patimah; Rahmat, Asmah

2014-01-01

The aim of this study was to examine the effects of extraction methods on antioxidant capacities of red dragon fruit peel and flesh. Antioxidant capacities were measured using ethylenebenzothiozoline-6-sulfonic acid (ABTS) radical cation assay and ferric reducing antioxidant power assay (FRAP). Total phenolic content (TPC) was determined using Folin-Ciocalteu reagent while quantitative determination of total flavonoid content (TFC) was conducted using aluminium trichloride colorimetric method. Betacyanin content (BC) was measured by spectrophotometer. Red dragon fruit was extracted using conventional (CV) and ultrasonic-assisted extraction (UE) technique to determine the most efficient way of extracting its antioxidant components. Results indicated that UE increased TFC, reduced the extraction yield, BC, and TPC, but exhibited the strongest scavenging activity for the peel of red dragon fruit. In contrast, UE reduced BC, TFC, and scavenging activity but increased the yield for the flesh. Nonetheless, UE slightly increases TPC in flesh. Scavenging activity and reducing power were highly correlated with phenolic and flavonoid compounds. Conversely, the scavenging activity and reducing power were weakly correlated with betacyanin content. This work gives scientific evidences for the consideration of the type of extraction techniques for the peel and flesh of red dragon fruit in applied research and food industry. PMID:25379555

Application of response surface methodology to optimise microbial inactivation of shrimp and conch by supercritical carbon dioxide.

PubMed

Chen, Manhua; Sui, Xiao; Ma, Xixiu; Feng, Xiaomei; Han, Yuqian

2015-03-30

Supercritical carbon dioxide (SC-CO2 ) has been shown to have a good pasteurising effect on food. However, very few research papers have investigated the possibility to exploit this treatment for solid foods, particularly for seafood. Considering the microbial safety of raw seafood consumption, the study aimed to explore the feasibility of microbial inactivation of shrimp (Metapenaeus ensis) and conch (Rapana venosa) by SC-CO2 treatment. Response surface methodology (RSM) models were established to predict and analyse the SC-CO2 process. A 3.69-log reduction in the total aerobic plate count (TPC) of shrimp was observed by SC-CO2 treatment at 53°C, 15 MPa for 40 min, and the logarithmic reduction in TPC of conch was 3.31 at 55°C, 14 MPa for 42 min. Sensory scores of the products achieved approximately 8 (desirable). The optimal parameters for microbial inactivation of shrimp and conch by SC-CO2 might be 55°C, 15 MPa and 40 min. SC-CO2 exerted a strong bactericidal effect on the TPC of shrimp and conch, and the products maintained good organoleptic properties. This study verified the feasibility of microbial inactivation of shrimp and conch by SC-CO2 treatment. © 2014 Society of Chemical Industry.

Antioxidant properties of chemical extracts and bioaccessible fractions obtained from six Spanish monovarietal extra virgin olive oils: assays in Caco-2 cells.

PubMed

Borges, Thays H; Cabrera-Vique, Carmen; Seiquer, Isabel

2015-07-01

The antioxidant activity and the total phenolic content (TPC) of six Spanish commercial monovarietal extra virgin olive oils (Arbequina, Cornicabra, Hojiblanca, Manzanilla, Picual and Picudo) were evaluated in chemical extracts and in bioaccessible fractions (BF) obtained after in vitro digestion. Moreover, the effects of the BF on cell viability and the generation of reactive oxygen species (ROS) were investigated in Caco-2 cell cultures. The in vitro digestion process increased the TPC and antioxidant activity evaluated by different methods (ABTS, DPPH and FRAP) compared with chemical extracts. After digestion, the Picual variety showed better beneficial effects in preserving cell integrity than the other varieties studied. Significant reductions of ROS production were observed after incubation of Caco-2 cells with the BF of all the varieties and, moreover, a protective effect against the oxidative stress induced by t-BOOH was shown for Arbequina, Cornicabra, Hojiblanca, Manzanilla and Picual. These findings seem to be an additional reason supporting the health benefits of Spanish extra virgin olive oil varieties. Multivariate factor analysis and principal component analysis were applied to assess the contribution of antioxidant activity and TPC, before and after digestion, to the characterization of the different varieties.

Comparison of total phenolic content and antioxidant activity of Kappaphycus alvarezii from Langkawi and Semporna

NASA Astrophysics Data System (ADS)

Mohamed, Norhidayu; Abdullah, Aminah

2016-11-01

The total phenolic content and antioxidant activity of Kappaphycus alvarezii obtained from Langkawi, Kedah and Semporna, Sabah were evaluated. The total phenolic content (TPC) of the extracts were determined according to the Folin Ciocalteau method and results were expressed as gallic acid equivalents. The antioxidant activities of the extracts were determined by three methods namely Free Radical Scavenging Activity (DPPH), Ferric Reducing Antioxidant Power (FRAP) and Trolox Equivalent Antioxidant Capacity (TEAC). Both of the TPC and FRAP assays showed that seaweed from Semporna, Sabah significantly (p<0.05) had higher antioxidant activities compared to Langkawi, Kedah one (Semporna's seaweed: 73.25 mg GAE/100g and 16.94 µmol TE/100g, Langkawi's seaweed: 54.35 mg GAE/100g and 10.01 µmol TE/100 g). However, the seaweed from Langkawi (60.93 µmol TE/100g) show higher TEAC value compared to seaweed from Semporna (36.36 µmol TE/100g) but for DPPH assay there was no significant difference (p>0.05) between the samples. Pearson coefficient correlation test, showed that there was a positive correlation (p<0.01) between TPC and antioxidant activity (FRAP assay) (r=0.980) and thus it can be concluded that the phenolic compounds was a contributor of the antioxidant activity in Kappaphycus alvarezii.

Effect of climate change on phytochemical diversity, total phenolic content and in vitro antioxidant activity of Aloe vera (L.) Burm.f.

PubMed

Kumar, Sandeep; Yadav, Amita; Yadav, Manila; Yadav, Jaya Parkash

2017-01-25

The aim of the present study was to analyse the effect of climate change on phytochemicals, total phenolic content (TPC) and antioxidant potential of methanolic extracts of Aloe vera collected from different climatic zones of the India. Crude methanolic extracts of A. vera from the different states of India were screened for presence of various phytochemicals, total phenolic content and in vitro antioxidant activity. Total phenolic content was tested by Folin-Ciocalteau reagent based assay whilst DPPH free radical scavenging assay, metal chelating assay, hydrogen peroxide scavenging assay, reducing power assay and β carotene-linoleic assay were used to assess the antioxidant potential of A. vera methanolic leaf extracts. Alkaloids, phenols, flavonoids, saponins, and terpenes were the main phytochemicals presents in all accessions. A significant positive correlation was found between TPC and antioxidant activity of different accessions. Extracts of highland and semi-arid zones possessed maximum antioxidant potential. Accessions from tropical zones showed the least antioxidant activity in all assays. It could be concluded that different agro-climatic conditions have effects on the phytochemicals, total phenolic content (TPC) and antioxidant potential of the A. vera plant. The results reveal that A. vera can be a potential source of novel natural antioxidant compounds.

Design of co-crystals/salts of some Nitrogenous bases and some derivatives of thiophene carboxylic acids through a combination of hydrogen and halogen bonds.

PubMed

Jennifer, Samson Jegan; Muthiah, Packianathan Thomas

2014-01-01

The utility of N-heterocyclic bases to obtain molecular complexes with carboxylic acids is well studied. Depending on the solid state interaction between the N-heterocyclic base and a carboxylic acid a variety of neutral or ionic synthons are observed. Meanwhile, pyridines and pyrimidines have been frequently chosen in the area of crystal engineering for their multipurpose functionality. HT (hetero trimers) and LHT (linear heterotetramers) are the well known synthons that are formed in the presence of pyrimidines and carboxylic acids. Fourteen crystals involving various substituted thiophene carboxylic acid derivatives and nitrogenous bases were prepared and characterized by using single crystal X-ray diffraction. The 14 crystals can further be divided into two groups [1a-7a], [8b-14b] based on the nature of the nitrogenous base. Carboxylic acid to pyridine proton transfer has occurred in 3 compounds of each group. In addition to the commonly occurring hydrogen bond based pyridine/carboxylic acid and pyrimidine/carboxylic acid synthons which is the reason for assembly of primary motifs, various other interactions like Cl…Cl, Cl…O, C-H…Cl, C-H…S add additional support in organizing these supermolecules into extended architectures. It is also interesting to note that in all the compounds π-π stacking occurs between the pyrimidine-pyrimidine or pyridine-pyridine or acid-acid moieties rather than acid-pyrimidine/pyridine. In all the compounds (1a-14b) either neutral O-H…Npyridyl/pyrimidine or charge-assisted Npyridinium-H…Ocarboxylate hydrogen bonds are present. The HT (hetero trimers) and LHT (linear heterotetramers) are dominant in the crystal structures of the adducts containing N-heterocyclic bases with two proton acceptors (1a-7a). Similar type supramolecular ladders are observed in 5TPC44BIPY (8b), TPC44BIPY (9b), TPC44TMBP (11b). Among the seven compounds [8b-14b] the extended ligands are linear in all except for the TMBP (10b, 11b, 12b). The structure of each compound depends on the dihedral angle between the carboxyl group and the nitrogenous base. All these compounds indicate three main synthons that regularly occur, namely linear heterodimer (HD), heterotrimer (HT) and heterotetramer (LHT).

Identification of a Novel C-Terminal Truncated WT1 Isoform with Antagonistic Effects against Major WT1 Isoforms

PubMed Central

Tatsumi, Naoya; Hojo, Nozomi; Sakamoto, Hiroyuki; Inaba, Rena; Moriguchi, Nahoko; Matsuno, Keiko; Fukuda, Mari; Matsumura, Akihide; Hayashi, Seiji; Morimoto, Soyoko; Nakata, Jun; Fujiki, Fumihiro; Nishida, Sumiyuki; Nakajima, Hiroko; Tsuboi, Akihiro; Oka, Yoshihiro; Hosen, Naoki; Sugiyama, Haruo; Oji, Yusuke

2015-01-01

The Wilms’ tumor gene WT1 consists of 10 exons and encodes a zinc finger transcription factor. There are four major WT1 isoforms resulting from alternative splicing at two sites, exon 5 (17AA) and exon 9 (KTS). All major WT1 isoforms are overexpressed in leukemia and solid tumors and play oncogenic roles such as inhibition of apoptosis, and promotion of cell proliferation, migration and invasion. In the present study, a novel alternatively spliced WT1 isoform that had an extended exon 4 (designated as exon 4a) with an additional 153 bp (designated as 4a sequence) at the 3’ end was identified and designated as an Ex4a(+)WT1 isoform. The insertion of exon 4a resulted in the introduction of premature translational stop codons in the reading frame in exon 4a and production of C-terminal truncated WT1 proteins lacking zinc finger DNA-binding domain. Overexpression of the truncated Ex4a(+)WT1 isoform inhibited the major WT1-mediated transcriptional activation of anti-apoptotic Bcl-xL gene promoter and induced mitochondrial damage and apoptosis. Conversely, suppression of the Ex4a(+)WT1 isoform by Ex4a-specific siRNA attenuated apoptosis. These results indicated that the Ex4a(+)WT1 isoform exerted dominant negative effects on anti-apoptotic function of major WT1 isoforms. Ex4a(+)WT1 isoform was endogenously expressed as a minor isoform in myeloid leukemia and solid tumor cells and increased regardless of decrease in major WT1 isoforms during apoptosis, suggesting the dominant negative effects on anti-apoptotic function of major WT1 isoforms. These results indicated that Ex4a(+)WT1 isoform had an important physiological function that regulated oncogenic function of major WT1 isoforms. PMID:26090994

Anti-oxidant activity and major chemical component analyses of twenty-six commercially available essential oils.

PubMed

Wang, Hsiao-Fen; Yih, Kuang-Hway; Yang, Chao-Hsun; Huang, Keh-Feng

2017-10-01

This study analyzed 26 commercially available essential oils and their major chemical components to determine their antioxidant activity levels by measuring their total phenolic content (TPC), reducing power (RP), β-carotene bleaching (BCB) activity, trolox equivalent antioxidant capacity (TEAC), and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging (DFRS) ability. The clove bud and thyme borneol essential oils had the highest RP, BCB activity levels, and TPC values among the 26 commercial essential oils. Furthermore, of the 26 essential oils, the clove bud and ylang ylang complete essential oils had the highest TEAC values, and the clove bud and jasmine absolute essential oils had the highest DFRS ability. At a concentration of 2.5 mg/mL, the clove bud and thyme borneol essential oils had RP and BCB activity levels of 94.56% ± 0.06% and 24.64% ± 0.03% and 94.58% ± 0.01% and 89.33% ± 0.09%, respectively. At a concentration of 1 mg/mL, the clove bud and thyme borneol essential oils showed TPC values of 220.00 ± 0.01 and 69.05 ± 0.01 mg/g relative to gallic acid equivalents, respectively, and the clove bud and ylang ylang complete essential oils had TEAC values of 809.00 ± 0.01 and 432.33 ± 0.01 μM, respectively. The clove bud and jasmine absolute essential oils showed DFRS abilities of 94.13% ± 0.01% and 78.62% ± 0.01%, respectively. Phenolic compounds of the clove bud, thyme borneol and jasmine absolute essential oils were eugenol (76.08%), thymol (14.36%) and carvacrol (12.33%), and eugenol (0.87%), respectively. The phenolic compounds in essential oils were positively correlated with the RP, BCB activity, TPC, TEAC, and DFRS ability. Copyright © 2017. Published by Elsevier B.V.

Proximate composition, phenolic content and in vitro antioxidant activity of aqueous extracts of the seaweeds Ascophyllum nodosum, Bifurcaria bifurcata and Fucus vesiculosus. Effect of addition of the extracts on the oxidative stability of canola oil under accelerated storage conditions.

PubMed

Agregán, Rubén; Munekata, Paulo E; Domínguez, Ruben; Carballo, Javier; Franco, Daniel; Lorenzo, José M

2017-09-01

Extracts from three macroalgae species (Ascophyllum nodosum (ANE), Bifurcaria bifurcata (BBE) and Fucus vesiculosus (FVE)) were tested for proximate composition (total solid, protein and total carbohydrate contents), total phenols content (TPC), and for their antioxidant activities in vitro in comparison to that of BHT compound by using four different assays (ABTS radical cation decolouration, DPPH free radical scavenging activity, ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC)). The inclusion of the extracts as oil stabilizers in canola oil in substitution of the synthetic antioxidant (BHT) was also evaluated by assessing lipid oxidation parameters (peroxide value (PV), p-anisidine value (AV), TBARS value, conjugated dienes (CD) and TOTOX index) under accelerated storage conditions (16days, 60°C). There was an inverse relationship between total solid content and total polyphenols content in the seaweed extracts. FVE showed an intermediate TPC (1.15g PGE/100g extract), but it presented the highest in vitro antioxidant activity when measured using the ABTS, DPPH and FRAP tests. BBE, that displayed the highest TPC (1.99g PGE/100g extract), only showed the highest in vitro antioxidant activity when measured using the ORAC test. ANE showed the lowest TPC and the lowest antioxidant activity in all the tests performed. The seaweed extracts added in a 500ppm concentration significantly reduced the oxidation during canola oil storage at 60°C, being this antioxidant effect significantly higher than that of BHT added at 50ppm. Results indicate that seaweed extracts can effectively inhibit the oxidation of canola oil and they can be a healthier alternative to the synthetic antioxidants in the oil industry. Copyright © 2016 Elsevier Ltd. All rights reserved.

In vitro anticancer activity of ethanolic extracts of Piper nigrum against colorectal carcinoma cell lines.

PubMed

Prashant, Akila; Rangaswamy, Chandini; Yadav, Anshu Kumar; Reddy, Varun; Sowmya, M N; Madhunapantula, Subbarao

2017-01-01

Piper nigrum (PN) is well known for its cytotoxic and pharmacological benefits. However, there is minimal documented evidence about its cytotoxic efficacy against colorectal carcinoma. We therefore sought to procure a precisely quantitative and qualitative result, pertaining the efficacy of an ethanolic extract of PN (EEPN) against colorectal carcinoma. EEPN was prepared by subjecting dried PN seeds to gradient ethanol fractionation. The total phenol content (TPC), antioxidant activity (AOA), and anti-inflammatory activity (AIA) were determined using Folin-Ciocalteu assay, ferric reducing ability of plasma and 2, 2-diphenyl-1-picrylhydrazyl methods, and human red blood cells membrane stabilizing assay, respectively. Colorectal carcinoma cell lines (HCT-116, HCT-15, and HT-29) were procured from National Centre for Cell Science, Pune, and were cultured in Dulbecco's modified eagle media supplemented with 10% fetal bovine serum and 1 mM L-glutamine. Cells were seeded into a 96-well plate, followed by treatment with increasing concentrations of EEPN. The cytotoxic efficacy was evaluated based on percentage inhibition of cells, using sulforhodamine-B assay. The IC-50 values were calculated using Prism software (Prism from GraphPad software, Inc. CA, USA). Biochemical analysis revealed that 50% EEPN exhibited higher TPC, AOA, and AIA when compared to 70% and 100% EEPN at any given concentration ( P = 0.041). Cytotoxic analysis revealed a dose-dependent response with maximum cellular inhibition at TPC of 6 and 3 μg/ml, using 50% EEPN. However, 50% inhibition of cellular growth using 50% EEPN was seen with TPC of 3.2, 2.9, and 1.9 μg/ml at 24, 48, and 72 h, respectively, in HCT-15 cells. Hence, time- and dose-dependent increase in the cytotoxic efficacy of 50% EEPN against colorectal carcinoma cell lines were noted ( P < 0.001). Given the significantly positive correlations exhibited between the biochemical and the cytotoxic properties evaluated in our study, we hereby conclude PN as a novel therapeutic spice for the treatment of colorectal carcinoma.

ANALYSIS OF PHENOLIC AND FLAVONOIDS OF WILD EPHEDRA ALATA PLANT EXTRACTS BY LC/PDA AND LC/MS AND THEIR ANTIOXIDANT ACTIVITY

PubMed Central

Al-Rimawi, Fuad; Abu-Lafi, Saleh; Abbadi, Jehad; Alamarneh, Ayman A.A.; Sawahreh, Raneen A.; Odeh, Imad

2017-01-01

Background: Ephedra is among Palestinian medicinal plants that are traditionally used in folkloric medicine for treating many diseases. Ephedra is known to have antibacterial and antioxidant effects. The goal of this study is to evaluate the antioxidant activity of different extracts from the Ephedra alata plant growing wild in Palestine, and to analyze their phenolic and flavonoid constituents by HPLC/PDA and HPLC/MS. Materials and Methods: Samples of the Ephedra alata plant grown wild in Palestine were extracted with three different solvents namely, 100% water, 80% ethanol, and 100% ethanol. The extracts were analyzed for their total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity (AA), as well as phenolic and flavonoids content by HPLC/PDA/MS. Results: The results revealed that the polarity of the extraction solvent affects the TPC, TFC, and AA of extracts. It was found that both TPC and AA are highest for plant extracted with 80% ethanol, followed by 100% ethanol, and finally with 100% water. TFC however was highest in the following order: 100% ethanol > 80% ethanol > water. Pearson correlation indicated that there is a significant correlation between AA and TPC, but there is no correlation between AA and TFC. Simultaneous HPLC-PDA and UHPLC-MS analysis of the ethanolic plant extracts revealed the presence of Luteolin-7-O-glucuronide flavone, Myricetin 3-rhamnoside and some other major polyphenolic compounds that share myricetin skeleton. Conclusion: Ephedra alata extract is rich in potent falvonoid glycosidic compounds as revealed by their similar overlaid UV-Vis spectra and UHPLC-MS results. On the basis of these findings, it is concluded that Ephedra alata constitutes a natural source of potent antioxidants that may prevent many diseases and could be potentially used in food, cosmetics, and pharmaceutical products. PMID:28573229

Improvement of Chia Seeds with Antioxidant Activity, GABA, Essential Amino Acids, and Dietary Fiber by Controlled Germination Bioprocess.

PubMed

Gómez-Favela, Mario Armando; Gutiérrez-Dorado, Roberto; Cuevas-Rodríguez, Edith Oliva; Canizalez-Román, Vicente Adrián; Del Rosario León-Sicairos, Claudia; Milán-Carrillo, Jorge; Reyes-Moreno, Cuauhtémoc

2017-12-01

Chia (Salvia hispanica L.) plant is native from southern Mexico and northern Guatemala. Their seeds are a rich source of bioactive compounds which protect consumers against chronic diseases. Germination improves functionality of the seeds due to the increase in the bioactive compounds and associated antioxidant activity. The purpose of this study was to obtain functional flour from germinated chia seeds under optimized conditions with increased antioxidant activity, phenolic compounds, GABA, essential amino acids, and dietary fiber with respect to un-germinated chia seeds. The effect of germination temperature and time (GT = 20-35 °C, Gt = 10-300 h) on protein, lipid, and total phenolic contents (PC, LC, TPC, respectively), and antioxidant activity (AoxA) was analyzed by response surface methodology as optimization tool. Chia seeds were germinated inside plastic trays with absorbent paper moisturized with 50 mL of 100 ppm sodium hypochlorite dissolution. The sprouts were dried (50 °C/8 h) and ground to obtain germinated chia flours (GCF). The prediction models developed for PC, LC, TPC, and AoxA showed high coefficients of determination, demonstrating their adequacy to explain the variations in experimental data. The highest values of PC, LC, TPC, and AoxA were obtained at two different optimal conditions (GT = 21 °C/Gt = 157 h; GT = 33 °C/Gt = 126 h). Optimized germinated chia flours (OGCF) had higher PC, TPC, AoxA, GABA, essential amino acids, calculated protein efficiency ratio (C-PER), and total dietary fiber (TDF) than un-germinated chia seed flour. The OGCF could be utilized as a natural source of proteins, dietary fiber, GABA, and antioxidants in the development of new functional beverages and foods.

Human visual function in the North Carolina clinical study on possible estuary-associated syndrome.

PubMed

Hudnell, H K; House, D; Schmid, J; Koltai, D; Stopford, W; Wilkins, J; Savitz, D A; Swinker, M; Music, S

2001-04-20

The U.S. Environmental Protection Agency assisted the North Carolina Department of Health and Human Services in conducting a study to investigate the potential for an association between fish kills in the North Carolina estuary system and the risk for persistent health effects. Impetus for the study was recent evidence suggesting that estuarine dinoflagellates, including members of the toxic Pfiesteria complex (TPC), P. piscicida and P. schumwayae, may release a toxin(s) that kills fish and adversely affects human health. This report describes one component of the study in which visual system function was assessed. Participants working primarily in estuaries inhabited by TPC or in off-shore waters thought not to contain TPC were studied. The potentially exposed estuary (n = 22) and unexposed offshore (n = 20) workers were matched for age, gender, and education. Visual acuity did not differ significantly between the cohorts, but visual contrast sensitivity (VCS), an indicator of visual pattern-detection ability for stimuli of various sizes, was significantly reduced by about 30% in the estuary relative to the offshore cohort. A further analysis that excluded participants having a history possibly predictive of neuropsychological impairment showed a similar VCS reduction. Additional analyses indicated that differences between the cohorts in age, education, smoking, alcohol consumption, and total time spent on any water did not account for the difference in VCS. Exploratory analyses suggested a possible association between the magnitude of VCS reduction and hours spent in contact with a fish kill. The profile of VCS deficit across stimulus sizes resembled that seen in organic solvent-exposed workers, but an assessment of occupational solvent, and other neurotoxicant, exposures did not indicate differences between the cohorts. These results suggest that factor(s) associated with the North Carolina estuaries, including the possibility of exposure to TPC toxin(s), may impair visual system function.

Efficacy of low-pressure foam cleaning compared to conventional cleaning methods in the removal of bacteria from surfaces associated with convenience food.

PubMed

Lambrechts, A A; Human, I S; Doughari, J H; Lues, J F R

2014-09-01

Food borne illnesses and food poisoning are cause for concern globally. The diseases are often caused by food contamination with pathogenic bacteria due largely to poor sanitary habits or storage conditions. Prevalence of some bacteria on cleaned and sanitised food contact surfaces from eight convenience food plants in Gauteng (South Africa) was investigated with the view to evaluate the efficacy of the cleaning methods used with such food contact surfaces. The microbial load of eight convenience food manufacturing plants was determined by sampling stainless steel food contact surfaces after they had been cleaned and sanitised at the end of a day's shift. Samples were analysed for Total Plate Count (TPC), Escherichia coli, Salmonella species, Staphylococcus aureus and Listeria species. Results showed that 59 % of the total areas sampled for TPC failed to comply with the legal requirements for surfaces, according to the Foodstuffs, Cosmetics and Disinfectants Act (< 100 cfu.cm(-2)). S. aureus and Salmonella were not detected, but Listeria was detected in 23 % and E. coli in 1.3 % of the samples. Fifty percent (50 %) of the plants applied conventional cleaning methods for cleaning and sanitation and 50 % used the low-pressure foam (LPF) method. There was significant difference (P ≤ 0.05) between the mean TPC values of the conventional cleaning method (14 358.82) compared to that of LPF method (2 386.51) but no significant difference (P > 0.05) in terms of Listeria species isolates obtained from both cleaning methods. The LPF method proved to be the superior cleaning option for lowering TPC counts. Regardless of cleaning method used, pathogens continued to flourish on various surfaces, including dry stainless steel, posing a contamination hazard for a considerable period depending on the contamination level and type of pathogen. Intensive training for proper chemical usage and strict procedural compliance among workers for efficient cleaning procedures is recommended.

Stability of total phenolic concentration and antioxidant capacity of extracts from pomegranate co-products subjected to in vitro digestion.

PubMed

Fawole, Olaniyi Amos; Opara, Umezuruike Linus

2016-09-13

Co-products obtained from pomegranate juice processing contain high levels of polyphenols with potential high added values. From value-addition viewpoint, the aim of this study was to evaluate the stability of polyphenolic concentrations in pomegranate fruit co-products in different solvent extracts and assess the effect on the total antioxidant capacity using the FRAP, DPPH˙ and ABTS(+) assays during simulated in vitro digestion. Pomegranate juice, marc and peel were extracted in water, 50 % ethanol (50%EtOH) and absolute ethanol (100%EtOH) and analysed for total phenolic concentration (TPC), total flavonoids concentration (TFC) and total antioxidant capacity in DPPH˙, ABTS(+) and FRAP assays before and after in vitro digestion. Total phenolic concentration (TPC) and total flavonoid concentration (TFC) were in the order of peel > marc > juice throughout the in vitro digestion irrespective of the extraction solvents used. However, 50 % ethanol extracted 1.1 to 12-fold more polyphenols than water and ethanol solvents depending on co-products. TPC and TFC increased significantly in gastric digests. In contrast, after the duodenal phase of in vitro digestion, polyphenolic concentrations decreased significantly (p < 0.05) compared to those obtained in gastric digests. Undigested samples and gastric digests showed strong and positive relationships between polyphenols and the antioxidant activities measured in DPPH, ABTS(+) and FRAP assays, with correlation coefficients (r (2)) ranging between 0.930-0.990. In addition, the relationships between polyphenols (TPC and TFC) and radical cation scavenging activity in ABTS(+) were moderately positive in duodenal digests. Findings from this study showed that concentration of pomegranate polyphenols and the antioxidant capacity during in vitro gastro-intestinal digestion may not reflect the pre-digested phenolic concentration. Thus, this study highlights the need to provide biologically relevant information on antioxidants by providing data reflecting their stability and activity after in vitro digestion.

Antioxidant and cytoprotective activities of Piper betle, Areca catechu, Uncaria gambir and betel quid with and without calcium hydroxide

PubMed Central

2013-01-01

Background Betel quid chewing is a popular habit in Southeast Asia. It is believed that chewing betel quid could reduce stress, strengthen teeth and maintain oral hygiene. The aim of this study was to investigate the antioxidant and cytoprotective activities of each of the ingredients of betel quid and compared with betel quid itself (with and without calcium hydroxide). The correlation of their cytoprotective and antioxidant activities with phenolic content was also determined. Methods Five samples (betel leaf, areca nut, gambir, betel quid and betel quid containing calcium hydroxide) were extracted in deionized distilled water for 12 hours at 37°C. Antioxidant activities were evaluated for radical scavenging activity using DPPH assay, ferric reducing activity using FRAP assay and lipid peroxidation inhibition activity using FTC assay. Total phenolic content (TPC) was determined using Folin-Ciocalteu procedure. Phenolic composition was analyzed using LC-MS/MS. Cytoprotective activity towards human gingival fibroblast cells was examined using MTT assay. Results Among the ingredients of betel quid, gambir demonstrated the highest antioxidant (DPPH - IC50 = 6.4 ± 0.8 μg/mL, FRAP - 5717.8 ± 537.6 μmol Fe(II)/mg), total phenolic content (TPC - 1142.5 ± 106.8 μg TAE/mg) and cytoprotective (100.1 ± 4.6%) activities. Betel quid when compared with betel quid containing calcium hydroxide has higher antioxidant (DPPH - IC50 =59.4 ± 4.4 μg/mL, FRAP - 1022.2 ± 235.7 μmol Fe(II)/mg), total phenolic content (TPC - 140.0 ± 22.3 μg TAE/mg), and cytoprotective (113.5 ± 15.9%) activities. However, all of the five samples showed good lipid peroxidation inhibition compared to vitamin E. LC-MS/MS analysis revealed the presence of quinic acid as the major compound of gambir and betel quid. A positive correlation was observed between TPC and radical scavenging (r = 0.972), reducing power (r = 0.981) and cytoprotective activity (r = 0.682). Conclusions The betel quid has higher TPC, and antioxidant and cytoprotective activities than betel quid with calcium hydroxide. The quinic acid in betel quid may play an important role in the oral health protection. PMID:24330738

Modeling Phase-Aligned Gamma-Ray and Radio Millisecond Pulsar Light Curves

NASA Technical Reports Server (NTRS)

Venter, C.; Johnson, T.; Harding, A.

2012-01-01

Since the discovery of the first eight gamma-ray millisecond pulsars (MSPs) by the Fermi Large Area Telescope, this population has been steadily expanding. Four of the more recent detections, PSR J00340534, PSR J1939+2134 (B1937+21; the first MSP ever discovered), PSR J1959+2048 (B1957+20; the first discovery of a black widow system), and PSR J2214+3000, exhibit a phenomenon not present in the original discoveries: nearly phase-aligned radio and gamma-ray light curves (LCs). To account for the phase alignment, we explore models where both the radio and gamma-ray emission originate either in the outer magnetosphere near the light cylinder or near the polar caps. Using a Markov Chain Monte Carlo technique to search for best-fit model parameters, we obtain reasonable LC fits for the first three of these MSPs in the context of altitude-limited outer gap (alOG) and two-pole caustic (alTPC) geometries (for both gamma-ray and radio emission). These models differ from the standard outer gap (OG)/two-pole caustic (TPC) models in two respects: the radio emission originates in caustics at relatively high altitudes compared to the usual conal radio beams, and we allow both the minimum and maximum altitudes of the gamma-ray and radio emission regions to vary within a limited range (excluding the minimum gamma-ray altitude of the alTPC model, which is kept constant at the stellar radius, and that of the alOG model, which is set to the position-dependent null charge surface altitude). Alternatively, phase-aligned solutions also exist for emission originating near the stellar surface in a slot gap scenario (low-altitude slot gap (laSG) models). We find that the alTPC models provide slightly better LC fits than the alOG models, and both of these give better fits than the laSG models (for the limited range of parameters considered in the case of the laSG models). Thus, our fits imply that the phase-aligned LCs are likely of caustic origin, produced in the outer magnetosphere, and that the radio emission for these pulsars may come from close to the light cylinder. In addition, we were able to constrain the minimum and maximum emission altitudes with typical uncertainties of 30% of the light cylinder radius. Our results therefore describe a third gamma-ray MSP subclass, in addition to the two previously found by Venter et al.: those with LCs fit by standard OG/TPC models and those with LCs fit by pair-starved polar cap models.

The translation initiation factor 3 subunit eIF3K interacts with PML and associates with PML nuclear bodies

DOE Office of Scientific and Technical Information (OSTI.GOV)

Salsman, Jayme; Pinder, Jordan; Tse, Brenda

2013-10-15

The promyelocytic leukemia protein (PML) is a tumor suppressor protein that regulates a variety of important cellular processes, including gene expression, DNA repair and cell fate decisions. Integral to its function is the ability of PML to form nuclear bodies (NBs) that serve as hubs for the interaction and modification of over 90 cellular proteins. There are seven canonical isoforms of PML, which encode diverse C-termini generated by alternative pre-mRNA splicing. Recruitment of specific cellular proteins to PML NBs is mediated by protein–protein interactions with individual PML isoforms. Using a yeast two-hybrid screen employing peptide sequences unique to PML isoformmore » I (PML-I), we identified an interaction with the eukaryotic initiation factor 3 subunit K (eIF3K), and in the process identified a novel eIF3K isoform, which we term eIF3K-2. We further demonstrate that eIF3K and PML interact both in vitro via pull-down assays, as well as in vivo within human cells by co-immunoprecipitation and co-immunofluorescence. In addition, eIF3K isoform 2 (eIF3K-2) colocalizes to PML bodies, particularly those enriched in PML-I, while eIF3K isoform 1 associates poorly with PML NBs. Thus, we report eIF3K as the first known subunit of the eIF3 translation pre-initiation complex to interact directly with the PML protein, and provide data implicating alternative splicing of both PML and eIF3K as a possible regulatory mechanism for eIF3K localization at PML NBs. - Highlights: • The PML-I C-terminus, encoded by exon 9, interacts with translation factor eIF3K. • We identify a novel eIF3K isoform that excludes exon 2 (eIF3K-2). • eIF3K-2 preferentially associates with PML bodies enriched in PML-I vs. PML-IV. • Alternative splicing of eIF3K regulates association with PML bodies.« less

pPKCδ activates SC35 splicing factor during H9c2 myoblastic differentiation.

PubMed

Zara, Susi; Falconi, Mirella; Rapino, Monica; Zago, Michela; Orsini, Giovanna; Mazzotti, Giovanni; Cataldi, Amelia; Teti, Gabriella

2011-01-01

Although Protein Kinase C (PKC) isoforms' role in the neonatal and adult cardiac tissue development and ageing has been widely described "in vivo", the interaction of such enzymes with specific nuclear substrates needs to be investigated. The aim of our research has been the study of the expression, localization and interaction with the splicing factor SC35 of PKC isoforms (α, δ, ε, ζ) and their potential role in modulating the transcription machinery. H9c2 cells induced to myoblast differentiation in the presence of 1% Horse Serum (HS) have represented our experimental model. The expression of PKC isoforms, their distribution and interaction with SC35 have been evaluated by western blotting, co-immunoprecipitation and double gold immunolabeling for transmission and scanning electron microscopy. Our results show PKCδ as the most expressed isoform in differentiated cells. Surprisingly, the distribution of PKCδ and SC35 does not show any significant modification between 10%FBS and 1%HS treated samples and no co-localization is observed. Moreover the interaction between the phosphorylated form of PKCδ (pPKCδ) and SC35 increases, is distributed and co-localizes within the nucleus of differentiated H9c2. These data represent reasonable evidence of pPKCδ mediated SC35 splicing factor activation, suggesting its direct effect on transcription via interaction with the transcription machinery. Furthermore, this co-localization represents a crucial event resulting in downstream changes in transcription of components which determine the morphological modifications related to cardiomyoblast differentiated phenotype.

Characterization of Basigin Isoforms and the Inhibitory Function of Basigin-3 in Human Hepatocellular Carcinoma Proliferation and Invasion ▿

PubMed Central

Liao, Cheng-Gong; Kong, Ling-Min; Song, Fei; Xing, Jin-Liang; Wang, Long-Xin; Sun, Zhi-Jian; Tang, Hao; Yao, Hui; Zhang, Yang; Wang, Li; Wang, Yu; Yang, Xiang-Min; Li, Yu; Chen, Zhi-Nan

2011-01-01

Basigin, which has four isoforms, plays an important role in invasion of hepatocellular carcinoma (HCC). Detailed transcriptional regulation and functions of the basigin isoforms have not been reported except in the case of the predominant isoform basigin-2, which act as inducer of matrix metalloproteinases (MMPs). Here we determined that basigin-2, basigin-3, and basigin-4 were the most abundant transcript variants in human cell lines. GeneRacer PCR and luciferase reporter assays showed that basigin-3 and basigin-4 were initiated from an alternative promoter. Basigin-3 and basigin-4 were widely expressed in various normal human tissues at the mRNA level and were upregulated in HCC tissues compared to in normal tissues. Western blotting and confocal imaging showed that glycosylated basigin-3 and basigin-4 were expressed and localized to the plasma membrane. However, in cultured cell lines, only native basigin-3, and not basigin-4, was detected at protein level. Overexpression of basigin-3 inhibited HCC cell proliferation, MMP induction, and cell invasion in vitro and in vivo. Bimolecular fluorescence complementation assays and nuclear magnetic resonance (NMR) analysis indicated that basigin-3 interacted with basigin-2 to form hetero-oligomers. In conclusion, we systematically investigated the alternative splicing of basigin and found that basigin-3 could inhibit HCC proliferation and invasion, probably through interaction with basigin-2 as an endogenous inhibitor via hetero-oligomerization. PMID:21536654

Heterologous expression, refolding and functional characterization of two antifreeze proteins from Fragilariopsis cylindrus (Bacillariophyceae).

PubMed

Uhlig, Christiane; Kabisch, Johannes; Palm, Gottfried J; Valentin, Klaus; Schweder, Thomas; Krell, Andreas

2011-12-01

Antifreeze proteins (AFPs) provide protection for organisms subjected to the presence of ice crystals. The psychrophilic diatom Fragilariopsis cylindrus which is frequently found in polar sea ice carries a multitude of AFP isoforms. In this study we report the heterologous expression of two antifreeze protein isoforms from F. cylindrus in Escherichia coli. Refolding from inclusion bodies produced proteins functionally active with respect to crystal deformation, recrystallization inhibition and thermal hysteresis. We observed a reduction of activity in the presence of the pelB leader peptide in comparison with the GS-linked SUMO-tag. Activity was positively correlated to protein concentration and buffer salinity. Thermal hysteresis and crystal deformation habit suggest the affiliation of the proteins to the hyperactive group of AFPs. One isoform, carrying a signal peptide for secretion, produced a thermal hysteresis up to 1.53°C±0.53°C and ice crystals of hexagonal bipyramidal shape. The second isoform, which has a long preceding N-terminal sequence of unknown function, produced thermal hysteresis of up to 2.34°C±0.25°C. Ice crystals grew in form of a hexagonal column in presence of this protein. The different sequences preceding the ice binding domain point to distinct localizations of the proteins inside or outside the cell. We thus propose that AFPs have different functions in vivo, also reflected in their specific TH capability. Copyright © 2011 Elsevier Inc. All rights reserved.

Dependence of cross-bridge kinetics on myosin light chain isoforms in rabbit and rat skeletal muscle fibres.

PubMed

Andruchov, Oleg; Andruchova, Olena; Wang, Yishu; Galler, Stefan

2006-02-15

Cross-bridge kinetics underlying stretch-induced force transients was studied in fibres with different myosin light chain (MLC) isoforms from skeletal muscles of rabbit and rat. The force transients were induced by stepwise stretches (< 0.3% of fibre length) applied on maximally Ca2+-activated skinned fibres. Fast fibre types IIB, IID (or IIX) and IIA and the slow fibre type I containing the myosin heavy chain isoforms MHC-IIb, MHC-IId (or MHC-IIx), MHC-IIa and MHC-I, respectively, were investigated. The MLC isoform content varied within fibre types. Fast fibre types contained the fast regulatory MLC isoform MLC2f and different proportions of the fast alkali MLC isoforms MLC1f and MLC3f. Type I fibres contained the slow regulatory MLC isoform MLC2s and the slow alkali MLC isoform MLC1s. Slow MLC isoforms were also present in several type IIA fibres. The kinetics of force transients differed by a factor of about 30 between fibre types (order from fastest to slowest kinetics: IIB > IID > IIA > I). The kinetics of the force transients was not dependent on the relative content of MLC1f and MLC3f. Type IIA fibres containing fast and slow MLC isoforms were about 1.2 times slower than type IIA fibres containing only fast MLC isoforms. We conclude that while the cross-bridge kinetics is mainly determined by the MHC isoforms present, it is affected by fast and slow MLC isoforms but not by the relative content of MLC1f and MLC3f. Thus, the physiological role of fast and slow MLC isoforms in type IIA fibres is a fine-tuning of the cross-bridge kinetics.

Analysis Strategy and Selection Procedure for νμ Charged Current Inclusive Interactions using the ^0 Detector (P0D) and the Time Projection Chambers of the T2K Experiment

NASA Astrophysics Data System (ADS)

Reinherz-Aronis, Erez; Clifton, Alex; Das, Raj; Toki, Walter; Johnson, Robert; Marino, Alysia; Yuan, Tianlu

2013-04-01

νμ Charge-Current events are produced and collected by the Near Detectors (ND280) in the Tokai to Kamioka (T2K) experiment. This talk focuses on those interactions that are created in the Pi-Zero detector (PøD) and whose momentum is measured by the Time Projection Chambers (TPC). The description of the analysis event selection is presented which includes Data-Quality cuts, Beam Quality parameters, and Fiducial Volume boundaries which are applied on the beginning of the PøD track. In addition the matching procedure of a TPC track to a PøD track and the optimization of this procedure in presented.

Operation of a LAr-TPC equipped with a multilayer LEM charge readout

DOE PAGES

Baibussinov, B.; Centro, S.; Farnese, C.; ...

2018-03-01

A novel detector for ionization signals in a single phase LAr-TPC has been experimented in the ICARINO test facility at the INFN Laboratories in Legnaro. It is based on the adoption of a multilayer Large Electron Multiplier (LEM) replacing the traditional anodic wire arrays. Cosmic muon tracks were detected allowing the measurement of energy deposition and a first determination of the signal to noise ratio. The analysis of the recorded events thus demonstrated the 3D reconstruction capability of this device for ionizing events in liquid Argon. The collected fraction of ionization charge is close to about 90%, with signal tomore » noise ratio similar to that measured with more traditional wire chambers.« less

Studying Silicon Photomultipliers and Light Signal Acquisition for the SBND Experiment

NASA Astrophysics Data System (ADS)

Savard, Claire; SBND Collaboration

2017-01-01

The Short-Baseline Near Detector (SBND) is one of three Liquid Argon Time Projection Chamber (LArTPC) based detectors that will be used in the Short-Baseline Neutrino (SBN) program at Fermilab. SBN will study the neutrino-argon interaction and search for oscillations at short baseline. Light produced in neutrino interactions inside a LArTPC provides a precision measurement of the initial interaction time of the event, essential for differentiating non-beam-background from beam-based signal. I will discuss the light guide system for SBND, with an emphasis on the Silicon Photomultiplier (SiPM) readout and data acquisition. In particular, I will show results from testing and characterizing a candidate electronics board for reading out the SiPM signals.

Topological signature in the NEXT high pressure xenon TPC

NASA Astrophysics Data System (ADS)

Ferrario, Paola; NEXT Collaboration

2017-09-01

The NEXT experiment aims to observe the neutrinoless double beta decay of 136Xe in a high-pressure xenon gas TPC using electroluminescence to amplify the signal from ionization. One of the main advantages of this technology is the possibility to use the topology of events with energies close to Qββ as an extra tool to reject background. In these proceedings we show with data from prototypes that an extra background rejection factor of 24.3 ± 1.4 (stat.)% can be achieved, while maintaining an efficiency of 66.7 ± 1.% for signal events. The performance expected in NEW, the next stage of the experiment, is to improve to 12.9% ± 0.6% background acceptance for 66.9% ± 0.6% signal efficiency.

Study of the dE/dx resolution of a GEM Readout Chamber prototype for the upgrade of the ALICE TPC

NASA Astrophysics Data System (ADS)

Mathis, Andreas

2018-02-01

The ALICE Collaboration is planning a major upgrade of its central barrel detectors to be able to cope with the increased LHC luminosity beyond 2020. For the TPC, this implies a replacement of the currently used gated MWPCs (Multi-Wire Proportional Chamber) by GEM (Gas Electron Multiplier) based readout chambers. In order to prove, that the present particle identification capabilities via measurement of the specific energy loss are retained after the upgrade, a prototype of the ALICE IROC (Inner Readout Chamber) has been evaluated in a test beam campaign at the CERN PS. The dE/dx resolution of the prototype has been proven to be fully compatible with the current MWPCs.

Microwave-induced three-photon coherence of Rydberg atomic states

NASA Astrophysics Data System (ADS)

Kwak, Hyo Min; Jeong, Taek; Lee, Yoon-Seok; Moon, Han Seb

2016-12-01

We investigate the three-photon coherence (TPC) effects of the Rydberg state in a Doppler-broadened four-level ladder-type atomic system for the 5S1/2(F=3)-5P3/2(F‧=4)-50D5/2-51P3/2 transition of 85Rb atoms. Upon interaction of the Rydberg Rb atom of the ladder-type electromagnetically induced transparency (EIT) scheme with a resonant microwave (MW) field, we numerically analyze the spectral features of the Rydberg TPC from two viewpoints, Autler-Townes splitting (AT-splitting) of the Rydberg EIT and three-photon electromagnetically induced absorption (TPEIA). We determine the criterion to differentiate between AT-splitting of the Rydberg EIT and TPEIA in the Doppler-broadened ladder-type atomic system.

Operation of a LAr-TPC equipped with a multilayer LEM charge readout

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baibussinov, B.; Centro, S.; Farnese, C.

A novel detector for ionization signals in a single phase LAr-TPC has been experimented in the ICARINO test facility at the INFN Laboratories in Legnaro. It is based on the adoption of a multilayer Large Electron Multiplier (LEM) replacing the traditional anodic wire arrays. Cosmic muon tracks were detected allowing the measurement of energy deposition and a first determination of the signal to noise ratio. The analysis of the recorded events thus demonstrated the 3D reconstruction capability of this device for ionizing events in liquid Argon. The collected fraction of ionization charge is close to about 90%, with signal tomore » noise ratio similar to that measured with more traditional wire chambers.« less

Necessity and Effect of Combating Legionella pneumophila in Municipal Shower Systems

PubMed Central

Wiik, Ragnhild; Krøvel, Anne Vatland

2014-01-01

The objective was to obtain research-based, holistic knowledge about necessity and effect of practiced measures against L. pneumophila in municipal shower systems in Stavanger, Norway. The effects of hot water treatment and membrane-filtering were investigated and compared to no intervention at all. The studies were done under real-world conditions. Additionally, a surveillance pilot study of municipal showers in Stavanger was performed. The validity of high total plate count (TPC) as an indication of L. pneumophila was evaluated. A simplified method, named “dripping method”, for detection and quantification of L. pneumophila was developed. The sensitivity of the dripping method is 5 colony-forming units of L. pneumophila/ml. The transference of L. pneumophila from shower water to aerosols was studied. Interviews and observational studies among the stakeholders were done in order to identify patterns of communication and behavior in a Legionella risk perspective. No substantial effects of the measures against L. pneumophila were demonstrated, except for a distally placed membrane filter. No significant positive correlation between TPC and L. pneumophila concentrations were found. L. pneumophila serogroup 2–14 was demonstrated in 21% of the 29 buildings tested in the surveillance pilot. Relatively few cells of L. pneumophila were transferred from shower water to aerosols. Anxiety appeared as the major driving force in the risk governance of Legionella. In conclusion, the risk of acquiring Legionnaires' disease from municipal shower systems is evaluated as low and uncertain. By eliminating ineffective approaches, targeted Legionella risk governance can be practiced. Risk management by surveillance is evaluated as appropriate. PMID:25490721

Cellulose microfibril structure: inspirations from plant diversity

NASA Astrophysics Data System (ADS)

Roberts, A. W.

2018-03-01

Cellulose microfibrils are synthesized at the plasma membrane by cellulose synthase catalytic subunits that associate to form cellulose synthesis complexes. Variation in the organization of these complexes underlies the variation in cellulose microfibril structure among diverse organisms. However, little is known about how the catalytic subunits interact to form complexes with different morphologies. We are using an evolutionary approach to investigate the roles of different catalytic subunit isoforms in organisms that have rosette-type cellulose synthesis complexes.

Selective Erasure of Distinct Forms of Long-Term Synaptic Plasticity Underlying Different Forms of Memory in the Same Postsynaptic Neuron.

PubMed

Hu, Jiangyuan; Ferguson, Larissa; Adler, Kerry; Farah, Carole A; Hastings, Margaret H; Sossin, Wayne S; Schacher, Samuel

2017-07-10

Generalization of fear responses to non-threatening stimuli is a feature of anxiety disorders. It has been challenging to target maladaptive generalized memories without affecting adaptive memories. Synapse-specific long-term plasticity underlying memory involves the targeting of plasticity-related proteins (PRPs) to activated synapses. If distinct tags and PRPs are used for different forms of plasticity, one could selectively remove distinct forms of memory. Using a stimulation paradigm in which associative long-term facilitation (LTF) occurs at one input and non-associative LTF at another input to the same postsynaptic neuron in an Aplysia sensorimotor preparation, we found that each form of LTF is reversed by inhibiting distinct isoforms of protein kinase M (PKM), putative PRPs, in the postsynaptic neuron. A dominant-negative (dn) atypical PKM selectively reversed associative LTF, while a dn classical PKM selectively reversed non-associative LTF. Although both PKMs are formed from calpain-mediated cleavage of protein kinase C (PKC) isoforms, each form of LTF is sensitive to a distinct dn calpain expressed in the postsynaptic neuron. Associative LTF is blocked by dn classical calpain, whereas non-associative LTF is blocked by dn small optic lobe (SOL) calpain. Interfering with a putative synaptic tag, the adaptor protein KIBRA, which protects the atypical PKM from degradation, selectively erases associative LTF. Thus, the activity of distinct PRPs and tags in a postsynaptic neuron contribute to the maintenance of different forms of synaptic plasticity at separate inputs, allowing for selective reversal of synaptic plasticity and providing a cellular basis for developing therapeutic strategies for selectively reversing maladaptive memories. Copyright © 2017 Elsevier Ltd. All rights reserved.

A new haemocyanin in cuttlefish (Sepia officinalis) eggs: sequence analysis and relevance during ontogeny

PubMed Central

2014-01-01

Background Haemocyanin is the respiratory protein of most of the Mollusca. In cephalopods and gastropods at least two distinct isoforms are differentially expressed. However, their physiological purpose is unknown. For the common cuttlefish Sepia officinalis, three isoforms are known so far, whereas for only two of them the complete mRNA sequences are available. In this study, we sequenced the complete mRNA of the third haemocyanin isoform and measured the relative expression of all three isoforms during embryogenesis to reveal a potential ontogenetic relevance. Results The cDNA of isoform 3 clearly correlates to the known Sepia officinalis haemocyanin subunits consisting of eight functional units and an internal duplicated functional unit d. Our molecular phylogenetic analyses reveal the third isoform representing a potentially ancestral haemocyanin isoform, and the analyses of the expression of haemocyanin type 3 reveal that haemocyanin type 3 only can be observed within eggs and during early development. Isoforms 1 and 2 are absent at these stages. After hatching, isoform 3 is downregulated, and isoform 1 and 2 are upregulated. Conclusions Our study clearly shows an embryonic relevance of the third isoform, which will be further discussed in the light of the changes in the physiological function of haemocyanin during ontogeny. Taken together with the fact that it could also be the isoform closest related to the common ancestor of cuttlefish haemocyanin, the phylogeny of cuttlefish haemocyanin may be recapitulated during its ontogeny. PMID:24499521

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schwab, Ryan S.; Ihnatovych, Ivanna; Yunus, Sharifah Z.S.A.

Myosin IC is a single headed member of the myosin superfamily that localizes to the cytoplasm and the nucleus, where it is involved in transcription by RNA polymerases I and II, intranuclear transport, and nuclear export. In mammalian cells, three isoforms of myosin IC are expressed that differ only in the addition of short isoform-specific N-terminal peptides. Despite the high sequence homology, the isoforms show differences in cellular distribution, in localization to nuclear substructures, and in their interaction with nuclear proteins through yet unknown mechanisms. In this study, we used EGFP-fusion constructs that express truncated or mutated versions of myosinmore » IC isoforms to detect regions that are involved in isoform-specific localization. We identified two nucleolar localization signals (NoLS). One NoLS is located in the myosin IC isoform B specific N-terminal peptide, the second NoLS is located upstream of the neck region within the head domain. We demonstrate that both NoLS are functional and necessary for nucleolar localization of specifically myosin IC isoform B. Our data provide a first mechanistic explanation for the observed functional differences between the myosin IC isoforms and are an important step toward our understanding of the underlying mechanisms that regulate the various and distinct functions of myosin IC isoforms. - Highlights: ► Two NoLS have been identified in the myosin IC isoform B sequence. ► Both NoLS are necessary for myosin IC isoform B specific nucleolar localization. ► First mechanistic explanation of functional differences between the isoforms.« less

Neuroblastoma differentiation involves the expression of two isoforms of the alpha-subunit of Go.

PubMed

Brabet, P; Pantaloni, C; Rodriguez, M; Martinez, J; Bockaert, J; Homburger, V

1990-04-01

The regulation of GTP-binding proteins (G proteins) was examined during the course of differentiation of neuroblastoma N1E-115 cells. N1E-115 cell membranes possess three Bordetella pertussis toxin (PTX) substrates assigned to alpha-subunits (G alpha) of Go (a G protein of unknown function) and "Gi (a G protein inhibitory to adenylate cyclase)-like" proteins and one substrate of Vibrio cholerae toxin corresponding to an alpha-subunit of Gs (a G protein stimulatory to adenylate cyclase). In undifferentiated cells, only one form of Go alpha was found, having a pI of 5.8 Go alpha content increased by approximately twofold from the undifferentiated state to 96 h of cell differentiation. This is mainly due to the appearance of another Go alpha form having a pI of 5.55. Both Go alpha isoforms have similar sizes on sodium dodecyl sulfate-polyacrylamide gels, are recognized by polyclonal antibodies to bovine brain Go alpha, are ADP-ribosylated by PTX, and are covalently myristylated in whole N1E-115 cells. In addition, immunofluorescent staining of N1E-115 cells with Go alpha antibodies revealed that association of Go alpha with the plasma membrane appears to coincide with the expression of the most acidic isoform and morphological cell differentiation. In contrast, the levels of both Gi alpha and Gs alpha did not significantly change, whereas that of the common beta-subunit increased by approximately 30% over the same period. These results demonstrate specific regulation of the expression of Go alpha during neuronal differentiation.

Regulation of human cardiac potassium channels by full-length KCNE3 and KCNE4.

PubMed

Abbott, Geoffrey W

2016-12-06

Voltage-gated potassium (Kv) channels comprise pore-forming α subunits and a multiplicity of regulatory proteins, including the cardiac-expressed and cardiac arrhythmia-linked transmembrane KCNE subunits. After recently uncovering novel, N-terminally extended (L) KCNE3 and KCNE4 isoforms and detecting their transcripts in human atrium, reported here are their functional effects on human cardiac Kv channel α subunits expressed in Xenopus laevis oocytes. As previously reported for short isoforms KCNE3S and KCNE4S, KCNE3L inhibited hERG; KCNE4L inhibited Kv1.1; neither form regulated the HCN1 pacemaker channel. Unlike KCNE4S, KCNE4L was a potent inhibitor of Kv4.2 and Kv4.3; co-expression of cytosolic β subunit KChIP2, which regulates Kv4 channels in cardiac myocytes, partially relieved Kv4.3 but not Kv4.2 inhibition. Inhibition of Kv4.2 and Kv4.3 by KCNE3L was weaker, and its inhibition of Kv4.2 abolished by KChIP2. KCNE3L and KCNE4L also exhibited subunit-specific effects on Kv4 channel complex inactivation kinetics, voltage dependence and recovery. Further supporting the potential physiological significance of the robust functional effects of KCNE4L on Kv4 channels, KCNE4L protein was detected in human atrium, where it co-localized with Kv4.3. The findings establish functional effects of novel human cardiac-expressed KCNE isoforms and further contribute to our understanding of the potential mechanisms influencing cardiomyocyte repolarization.

Genetic variation of the prion protein gene (PRNP) in alpaca (Vicugna pacos)

USDA-ARS?s Scientific Manuscript database

Transmissible spongiform encephalopathies (TSE) are caused by accumulation of a misfolded form of the prion protein (PrP). The normal cellular isoform of PrP is produced by the prion gene (PRNP) and is highly expressed in the central nervous system. Currently, there is an absence of information rega...

A transfectant RK13 cell line permissive to classical caprine scrapie prion propagation

USDA-ARS?s Scientific Manuscript database

Classical scrapie is a form of transmissible spongiform encephalopathies (TSE) affecting domestic goats and sheep and disease is characterized by the accumulation of abnormal conformational isoform (PrP-Sc) of normal cellular prion protein (PrP-C) in the central nervous system and, in most cases, ly...

The characterization and molecular structure of hepatoproliferin: a liver regeneration factor from rat hepatocytes.

PubMed

Oosthuizen, Mathys M J; Lambrechts, Hugo

2007-01-01

Hepatoproliferin (HPF) was purified from regenerating rat livers as an oligomeric entity (big-HPF) from which the monomeric form (small-HPF) could be obtained using disaggregating conditions. By using a solid-phase ion-exchange method, small-HPF was forced to dissociate into two charged ionic species, namely norepinephrine (NE) and a sulfonated disaccharide with a molecular structure consisting of D-glucuronic acid bound to glucosamine 2,6-disulfate by a beta-glycosidic linkage having a beta, 1 --> 4 configuration. Monomeric HPF stemmed from the formation of three electrostatic bonds between the protonated amine groups of three norepinephrines, of which two bind to the deprotonated sulfonic groups of glucosamine 2,6-disulfate and one to the deprotonated carboxylic group of glucuronic acid, to constitute a tightly associated complex with a molecular mass of 1046 Da. This represents one of the two purified isoforms of small-HPF. The other isoform, which has a lower molecular mass of 877 Da, lack one NE, leaving the weaker carboxylic group of glucuronic acid unoccupied, to constitute a more acidic form of HPF.

Novel function of STAT1beta in B cells: induction of cell death by a mechanism different from that of STAT1alpha.

PubMed

Najjar, Imen; Schischmanoff, Pierre Olivier; Baran-Marszak, Fanny; Deglesne, Pierre-Antoine; Youlyouz-Marfak, Ibtissam; Pampin, Mathieu; Feuillard, Jean; Bornkamm, Georg W; Chelbi-Alix, Mounira K; Fagard, Remi

2008-12-01

Alternate splicing of STAT1 produces two isoforms: alpha, known as the active form, and beta, previously shown to act as a dominant-negative factor. Most studies have dealt with STAT1alpha, showing its involvement in cell growth control and cell death. To examine the specific function of either isoform in cell death, a naturally STAT1-deficient human B cell line was transfected to express STAT1alpha or STAT1beta. STAT1alpha, expressed alone, enhanced cell death, potentiated the fludarabine-induced apoptosis, and enhanced the nuclear location, the phosphorylation, and the transcriptional activity of p53. Unexpectedly, STAT1beta, expressed alone, induced cell death through a mechanism that was independent of the nuclear function of p53. Indeed, in STAT1beta-expressing B cells, p53 was strictly cytoplasmic where it formed clusters, and there was no induction of the transcriptional activity of p53. These data reveal a novel role of STAT1beta in programmed cell death, which is independent of p53.

Tissue Factor Pathway Inhibitor: Multiple Anticoagulant Activities for a Single Protein.

PubMed

Mast, Alan E

2016-01-01

Tissue factor (TF) pathway inhibitor (TFPI) is an anticoagulant protein that inhibits early phases of the procoagulant response. Alternatively spliced isoforms of TFPI are differentially expressed by endothelial cells and human platelets and plasma. The TFPIβ isoform localizes to the endothelium surface where it is a potent inhibitor of TF-factor VIIa complexes that initiate blood coagulation. The TFPIα isoform is present in platelets. TFPIα contains a stretch of 9 amino acids nearly identical to those found in the B-domain of factor V that are well conserved in mammals. These amino acids provide exosite binding to activated factor V, which allows for TFPIα to inhibit prothrombinase during the initiation phase of blood coagulation. Endogenous inhibition at this point in the coagulation cascade was only recently recognized and has provided a biochemical rationale to explain the pathophysiological mechanisms underlying several clinical disorders. These include the east Texas bleeding disorder that is caused by production of an altered form of factor V with high affinity for TFPI and a paradoxical procoagulant effect of heparins. In addition, these findings have led to ideas for pharmacological targeting of TFPI that may reduce bleeding in hemophilia patients. © 2015 American Heart Association, Inc.

Identification of a novel higher molecular weight isoform of USP7/HAUSP that interacts with the Herpes simplex virus type-1 immediate early protein ICP0.

PubMed

Antrobus, Robin; Boutell, Chris

2008-10-01

The Herpes simplex virus type-1 (HSV-1) regulatory protein ICP0, a RING-finger E3 ubiquitin ligase, stimulates the onset of viral lytic replication and the reactivation of quiescent viral genomes from latency. Like many ubiquitin ligases ICP0 induces its own ubiquitination, a process that can lead to its proteasome-dependent degradation. ICP0 counteracts this activity by recruiting the cellular ubiquitin-specific protease USP7/HAUSP. Here we show that ICP0 can also interact with a previously unidentified isoform of USP7 (termed here USP7(beta)). This isoform is not a predominantly ubiquitinated, SUMO-modified, or phosphorylated species of USP7 but is constitutively expressed in a number of different cell types. Like USP7, USP7(beta) binds specifically to an electrophilic ubiquitin probe, indicating that it contains an accessible catalytic core with potential ubiquitin-protease activity. The interaction formed between ICP0 and USP7(beta) requires ICP0 to have an intact USP7-binding domain and results in its susceptibility to ICP0-mediated degradation during HSV-1 infection.

Construction of a laccase chimerical gene: recombinant protein characterization and gene expression via yeast surface display.

PubMed

Bleve, G; Lezzi, C; Spagnolo, S; Rampino, P; Perrotta, C; Mita, G; Grieco, Francesco

2014-03-01

The ERY4 laccase gene from Pleurotus eryngii was expressed in Saccharomyces cerevisiae and the recombinant laccase resulted to be not biologically active. This gene was thus modified to obtain chimerical enzymes derived from the substitution of N-, C- and both N- and C-terminal regions with the corresponding regions of Ery3 laccase, another laccase isoform of P. eryngii. The chimerical isoform named 4NC3, derived from the substitution of both N- and C-terminal regions, showed the best performances in terms of enzymatic activities, affinities for different substrates and stability at a broad range of temperatures and pHs. The chimerical 4NC3 laccase isoform was displayed on the cell surface of S. cerevisiae using the N-terminal fusion with either the Pir2 or the Flo1 S. cerevisiae proteins as anchor attachment sequence. Immunofluorescence microscopy and Western blot analyses confirmed the localization of 4NC3 on the yeast cell surface. The enzyme activity on specific laccase substrates revealed that 4NC3 laccase was immobilized in active form on the cell surface. To our knowledge, this is the first example of expression of a chimerical fungal laccase by yeast cell display.

Overexpression of high molecular weight FGF-2 forms inhibits glioma growth by acting on cell-cycle progression and protein translation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lemiere, Sylvie; University Bordeaux1, Talence, F-33405; Azar, Rania

2008-12-10

In order to clarify the role of HMW FGF-2 in glioma development and angiogenesis, we over-expressed different human FGF-2 isoforms in C6 rat glioma cell line using a tetracycline-regulated expression system. Phenotypic modifications were analyzed in vitro and compared to untransfected cells or to cells over-expressing 18 kDa FGF-2 or all FGF-2 isoforms. In particular, we demonstrate that HMW FGF-2 has unique features in inhibiting glioma cell proliferation. HMW FGF-2 expressing cells showed a cell-cycle arrest at the G2M, demonstrating a role of HMW FGF-2 in controlling the entry in mitosis. Moreover, hydroxyurea was ineffective in blocking cells at themore » G1S boundary when HMW FGF-2 was expressed. We also show that the HMW FGF-2 isoforms inhibit 4E-BP1 phosphorylation at critical sites restoring the translation inhibitory activity of 4E-BP1. In vivo, inhibition of tumor growth was observed when cells expressed HMW FGF-2. This indicates that HMW FGF-2 inhibits tumor growth in glioma cells by acting on cell-cycle progression and protein translation.« less

Presence of angiotensin converting enzyme isoforms in larval lepidoptera (Spodoptera littoralis).

PubMed

Lemeire, E; Van Camp, J; Smagghe, G

2007-01-01

In this research the presence of angiotensin converting enzyme (ACE) in larvae of the lepidopteran Spodoptera littoralis was evaluated. Making use of the substrate Abz-FRK-(Dnp)P-OH and the specific inhibitor captopril at 10 microM, ACE activity was determined in a fluorescence assay for intact larvae, hemolymph, head, midgut and dorsal tissue. In dorsal tissue and hemolymph, ACE activity was highest. These data are consistent with a possible role for ACE in contractions of the dorsal vessel and metabolism of circulating peptide hormones in the hemolymph. After the presence of ACE was confirmed, a sequential procedure of anion exchange and size exclusion chromatography was applied to purify ACE from whole wandering larvae (last stage). With this procedure, three different ACE pools were collected that cleaved the fluorogenic substrate Abz-FRK-(Dnp)P-OH. Activity could be inhibited by a final concentration of 2.5 microM captopril. In addition, two out of three samples eluted at different salt concentration and thus ACE 1, 2 and 3 represent at least two different ACE isoforms. These data reveal that ACE is present in S. littoralis and that at least two out of three isolated ACE forms are truly isoforms.

The β subunit of yeast AMP-activated protein kinase directs substrate specificity in response to alkaline stress.

PubMed

Chandrashekarappa, Dakshayini G; McCartney, Rhonda R; O'Donnell, Allyson F; Schmidt, Martin C

2016-12-01

Saccharomyces cerevisiae express three isoforms of Snf1 kinase that differ by which β subunit is present, Gal83, Sip1 or Sip2. Here we investigate the abundance, activation, localization and signaling specificity of the three Snf1 isoforms. The relative abundance of these isoforms was assessed by quantitative immunoblotting using two different protein extraction methods and by fluorescence microscopy. The Gal83 containing isoform is the most abundant in all assays while the abundance of the Sip1 and Sip2 isoforms is typically underestimated especially in glass-bead extractions. Earlier studies to assess Snf1 isoform function utilized gene deletions as a means to inactivate specific isoforms. Here we use point mutations in Gal83 and Sip2 and a 17 amino acid C-terminal truncation of Sip1 to inactivate specific isoforms without affecting their abundance or association with the other subunits. The effect of low glucose and alkaline stresses was examined for two Snf1 phosphorylation substrates, the Mig1 and Mig2 proteins. Any of the three isoforms was capable of phosphorylating Mig1 in response to glucose stress. In contrast, the Gal83 isoform of Snf1 was both necessary and sufficient for the phosphorylation of the Mig2 protein in response to alkaline stress. Alkaline stress led to the activation of all three isoforms yet only the Gal83 isoform translocates to the nucleus and phosphorylates Mig2. Deletion of the SAK1 gene blocked nuclear translocation of Gal83 and signaling to Mig2. These data strongly support the idea that Snf1 signaling specificity is mediated by localization of the different Snf1 isoforms. Copyright © 2016 Elsevier Inc. All rights reserved.

The β subunit of yeast AMP-activated protein kinase directs substrate specificity in response to alkaline stress

PubMed Central

Chandrashekarappa, Dakshayini G.; McCartney, Rhonda R.; O’Donnell, Allyson F.; Schmidt, Martin C.

2016-01-01

Saccharomyces cerevisiae express three isoforms of Snf1 kinase that differ by which β subunit is present, Gal83, Sip1 or Sip2. Here we investigate the abundance, activation, localization and signaling specificity of the three Snf1 isoforms. The relative abundance of these isoforms was assessed by quantitative immunoblotting using two different protein extraction methods and by fluorescence microscopy. The Gal83 containing isoform is the most abundant in all assays while the abundance of the Sip1 and Sip2 isoforms is typically underestimated especially in glass-bead extractions. Earlier studies to assess Snf1 isoform function utilized gene deletions as a means to inactivate specific isoforms. Here we use point mutations in Gal83 and Sip2 and a 17 amino acid C-terminal truncation of Sip1 to inactivate specific isoforms without affecting their abundance or association with the other subunits. The effect of low glucose and alkaline stresses was examined for two Snf1 phosphorylation substrates, the Mig1 and Mig2 proteins. Any of the three isoforms was capable of phosphorylating Mig1 in response to glucose stress. In contrast, the Gal83 isoform of Snf 1 was both necessary and sufficient for the phosphorylation of the Mig2 protein in response to alkaline stress. Alkaline stress led to the activation of all three isoforms yet only the Gal83 isoform translocates to the nucleus and phosphorylates Mig2. Deletion of the SAK1 gene blocked nuclear translocation of Gal83 and signaling to Mig2. These data strongly support the idea that Snf1 signaling specificity is mediated by localization of the different Snf1 isoforms. PMID:27592031

Differential toxicity of TDP-43 isoforms depends on their sub-mitochondrial localization in neuronal cells.

PubMed

Salvatori, Illari; Ferri, Alberto; Scaricamazza, Silvia; Giovannelli, Ilaria; Serrano, Alessia; Rossi, Simona; D'Ambrosi, Nadia; Cozzolino, Mauro; Di Giulio, Andrea; Moreno, Sandra; Valle, Cristiana; Carrì, Maria Teresa

2018-05-20

TAR DNA binding protein 43 (TDP-43) is an RNA binding protein and a major component of protein aggregates found in Amyotrophic Lateral Sclerosis and several other neurodegenerative diseases. TDP-43 exists as a full length protein and as two shorter forms of 25 and 35 kDa. Full length mutant TDP-43s found in ALS patients re-localize from the nucleus to the cytoplasm and in part to mitochondria, where they exert a toxic role associated with neurodegeneration. However, induction of mitochondrial damage by TDP-43 fragments is yet to be clarified. In this work, we show that the mitochondrial 35 kDa truncated form of TDP-43 is restricted to the intermembrane space while the full length forms also localise in the mitochondrial matrix in cultured neuronal NSC-34 cells. Interestingly, the full length forms clearly affect mitochondrial metabolism and morphology, possibly via their ability to inhibit the expression of Complex I subunits encoded by the mitochondrial-transcribed mRNAs, while the 35 kDa form does not. In the light of the known differential contribution of the full length and short isoforms to generate toxic aggregates, we propose that the presence of full length TDP-43s in the matrix is a primary cause of mitochondrial damage. This in turn may cause oxidative stress inducing toxic oligomers formation, in which short TDP-43 forms play a major role. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

One isoform of Arg/Abl2 tyrosine kinase is nuclear and the other seven cytosolic isoforms differently modulate cell morphology, motility and the cytoskeleton

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bianchi, Cristina; Torsello, Barbara; Di Stefano, Vitalba

The non-receptor tyrosine kinase Abelson related gene (Arg/Abl2) regulates cell migration and morphogenesis by modulating the cytoskeleton. Arg promotes actin-based cell protrusions and spreading, and inhibits cell migration by attenuating stress fiber formation and contractility via activation of the RhoA inhibitor, p190RhoGAP, and by regulating focal adhesion dynamics also via CrkII phosphorylation. Eight full-length Arg isoforms with different N- and C-termini are endogenously expressed in human cells. In this paper, the eight Arg isoforms, subcloned in the pFLAG-CMV2 vector, were transfected in COS-7 cells in order to study their subcellular distribution and role in cell morphology, migration and cytoskeletal modulation.more » The transfected 1BSCTS Arg isoform has a nuclear distribution and phosphorylates CrkII in the nucleus, whilst the other isoforms are detected in the cytoplasm. The 1BLCTL, 1BSCTL, 1ASCTS isoforms were able to significantly decrease stress fibers, induce cell shrinkage and filopodia-like protrusions with a significant increase in p190RhoGAP phosphorylation. In contrast, 1ALCTL, 1ALCTS, 1ASCTL and 1BLCTS isoforms do not significantly decrease stress fibers and induce the formation of retraction tail-like protrusions. The 1BLCTL and 1ALCTL isoforms have different effects on cell migration and focal adhesions. All these data may open new perspectives to study the mechanisms of cell invasiveness. -Highlights: • Each of the eight Arg isoforms was transfected in COS-7 cells. • Only the 1BSCTS Arg isoform has a nuclear distribution in transfected cells. • The cytoplasmic isoforms and F-actin colocalize cortically and in cell protrusions. • Arg isoforms differently phosphorylate p190RhoGAP and CrkII. • Arg isoforms differently modulate stress fibers, cell protrusions and motility.« less

Differential Properties of Cytomegalovirus pUL97 Kinase Isoforms Affect Viral Replication and Maribavir Susceptibility

PubMed Central

Webel, Rike; Hakki, Morgan; Prichard, Mark N.; Rawlinson, William D.; Marschall, Manfred

2014-01-01

ABSTRACT The human cytomegalovirus (HCMV)-encoded kinase pUL97 is required for efficient viral replication. Previous studies described two isoforms of pUL97, the full-length isoform (M1) and a smaller isoform likely resulting from translation initiation at codon 74 (M74). Here, we report the detection of a third pUL97 isoform during viral infection resulting from translation initiation at codon 157 (isoform M157). The consistent expression of isoform M157 as a minor component of pUL97 during infection with clinical and laboratory-adapted HCMV strains was suppressed when codon 157 was mutagenized. Viral mutants expressing specific isoforms were generated to compare their growth and drug susceptibility phenotypes, as well as pUL97 intracellular localization patterns and kinase activities. The exclusive expression of isoform M157 resulted in substantially reduced viral growth and resistance to the pUL97 inhibitor maribavir while retaining susceptibility to ganciclovir. Confocal imaging demonstrated reduced nuclear import of amino-terminal deletion isoforms compared to isoform M1. Isoform M157 showed reduced efficiency of various substrate protein interactions and autophosphorylation, whereas Rb phosphorylation was preserved. These results reveal differential properties of pUL97 isoforms that affect viral replication, with implications for the antiviral efficacy of maribavir. IMPORTANCE The HCMV UL97 kinase performs important functions in viral replication that are targeted by the antiviral drug maribavir. Here, we describe a naturally occurring short isoform of the kinase that when expressed by itself in a recombinant virus results in altered intracellular localization, impaired growth, and high-level resistance to maribavir compared to those of the predominant full-length counterpart. This is another factor to consider in explaining why maribavir appears to have variable antiviral activity in cell culture and in vivo. PMID:24522923

Simultaneous Detection of Human C-Terminal p53 Isoforms by Single Template Molecularly Imprinted Polymers (MIPs) Coupled with Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)-Based Targeted Proteomics.

PubMed

Jiang, Wenting; Liu, Liang; Chen, Yun

2018-03-06

Abnormal expression of C-terminal p53 isoforms α, β, and γ can cause the development of cancers including breast cancer. To date, much evidence has demonstrated that these isoforms can differentially regulate target genes and modulate their expression. Thus, quantification of individual isoforms may help to link clinical outcome to p53 status and to improve cancer patient treatment. However, there are few studies on accurate determination of p53 isoforms, probably due to sequence homology of these isoforms and also their low abundance. In this study, a targeted proteomics assay combining molecularly imprinted polymers (MIPs) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for simultaneous quantification of C-terminal p53 isoforms. Isoform-specific surrogate peptides (i.e., KPLDGEYFTLQIR (peptide-α) for isoform α, KPLDGEYFTLQDQTSFQK (peptide-β) for isoform β, and KPLDGEYFTLQMLLDLR (peptide-γ) for isoform γ) were first selected and used in both MIPs enrichment and mass spectrometric detection. The common sequence KPLDGEYFTLQ of these three surrogate peptides was used as single template in MIPs. In addition to optimization of imprinting conditions and characterization of the prepared MIPs, binding affinity and cross-reactivity of the MIPs for each surrogate peptide were also evaluated. As a result, a LOQ of 5 nM was achieved, which was >15-fold more sensitive than that without MIPs. Finally, the assay was validated and applied to simultaneous quantitative analysis of C-terminal p53 isoforms α, β, and γ in several human breast cell lines (i.e., MCF-10A normal cells, MCF-7 and MDA-MB-231 cancer cells, and drug-resistant MCF-7/ADR cancer cells). This study is among the first to employ single template MIPs and cross-reactivity phenomenon to select isoform-specific surrogate peptides and enable simultaneous quantification of protein isoforms in LC-MS/MS-based targeted proteomics.

The type III inositol 1,4,5-trisphosphate receptor preferentially transmits apoptotic Ca2+ signals into mitochondria.

PubMed

Mendes, Carolina C P; Gomes, Dawidson A; Thompson, Mayerson; Souto, Natalia C; Goes, Tercio S; Goes, Alfredo M; Rodrigues, Michele A; Gomez, Marcus V; Nathanson, Michael H; Leite, M Fatima

2005-12-09

There are three isoforms of the inositol 1,4,5- trisphosphate receptor (InsP(3)R), each of which has a distinct effect on Ca(2+) signaling. However, it is not known whether each isoform similarly plays a distinct role in the activation of Ca(2+)-mediated events. To investigate this question, we examined the effects of each InsP(3)R isoform on transmission of Ca(2+) signals to mitochondria and induction of apoptosis. Each isoform was selectively silenced using isoform-specific small interfering RNA in Chinese hamster ovary cells, which express all three InsP(3)R isoforms. ATP-induced cytosolic Ca(2+) signaling patterns were altered, regardless of which isoform was silenced, but in a different fashion depending on the isoform. ATP also induced Ca(2+) signals in mitochondria, which were inhibited more effectively by silencing the type III InsP(3)R than by silencing either the type I or type II isoform. The type III isoform also co-localized most strongly with mitochondria. When apoptosis was induced by activation of either the extrinsic or intrinsic apoptotic pathway, induction was reduced most effectively by silencing the type III InsP(3)R. These findings provide evidence that the type III isoform of the InsP(3)R plays a special role in induction of apoptosis by preferentially transmitting Ca(2+) signals into mitochondria.

Antioxidant Activity, Total Phenolics Content, Anthocyanin, and Color Stability of Isotonic Model Beverages Colored with Andes Berry (Rubus glaucus Benth) Anthocyanin Powder

PubMed Central

Estupiñan, D.C.; Schwartz, S.J.; Garzón, G.A.

2013-01-01

The stability of anthocyanin (ACN) freeze-dried powders from Andes berry (Rubus glaucus Benth) as affected by storage, addition of maltodextrin as a carrier agent, and illumination was evaluated in isotonic model beverages. The ethanolic ACN extract was freeze dried with and without maltodextrin DE 20. Isotonic model beverages were colored with freeze-dried ACN powder (FDA), freeze-dried ACN powder with maltodextrin (MFDA), and red nr 40. Beverages were stored in the dark and under the effect of illumination. Half life of the ACNs, changes in color, total phenolics content (TPC), and antioxidant activity were analyzed for 71 d. Addition of maltodextrin and absence of light stabilized the color of beverages and improved ACN and TPC stability during storage. The antioxidant activity of the beverages was higher when they were colored with MFDA and highly correlated with ACN content. There was no correlation between antioxidant activity and TPC. It is concluded that addition of maltodextrin DE 20 as a carrier agent during freeze-drying improves the color and stability of nutraceutical antioxidants present in Andes berry extract. This suggests a protective enclosing of ACNs within a maltodextrin matrix with a resulting powder that could serve as a supplement or additive to naturally color and to enhance the antioxidant capacity of isotonic beverages. PMID:21535712

Optimizing Extraction Conditions of Free and Bound Phenolic Compounds from Rice By-Products and Their Antioxidant Effects.

PubMed

Irakli, Maria; Kleisiaris, Fotis; Kadoglidou, Kalliopi; Katsantonis, Dimitrios

2018-06-13

Rice by-products are extensively abundant agricultural wastes from the rice industry. This study was designed to optimize experimental conditions for maximum recovery of free and bound phenolic compounds from rice by-products. Optimized conditions were determined using response surface methodology based on total phenolic content (TPC), ABTS radical scavenging activity and ferric reducing power (FRAP). A Box-Behnken design was used to investigate the effects of ethanol concentration, extraction time and temperature, and NaOH concentration, hydrolysis time and temperature for free and bound fractions, respectively. The optimal conditions for the free phenolics were 41⁻56%, 40 °C, 10 min, whereas for bound phenolics were 2.5⁻3.6 M, 80 °C, 120 min. Under these conditions free TPC, ABTS and FRAP values in the bran were approximately 2-times higher than in the husk. However, bound TPC and FRAP values in the husk were 1.9- and 1.2-times higher than those in the bran, respectively, while bran fraction observed the highest ABTS value. Ferulic acid was most evident in the bran, whereas p -coumaric acid was mostly found in the husk. Findings from this study demonstrates that rice by-products could be exploited as valuable sources of bioactive components that could be used as ingredients of functional food and nutraceuticals.

Vaccinium meridionale Swartz extracts and their addition in beef burgers as antioxidant ingredient.

PubMed

López-Padilla, Alexis; Martín, Diana; Villanueva Bermejo, David; Jaime, Laura; Ruiz-Rodriguez, Alejandro; Restrepo Flórez, Claudia Estela; Rivero Barrios, Diana Marsela; Fornari, Tiziana

2018-01-01

Vaccinium meridionale Swartz (mortiño) constitutes a source of bioactive phytochemicals, but reports related to its efficient and green production are scarce. In this study, pressurized liquid extraction (PLE) and ultrasound-assisted extraction of mortiño were compared. Total phenolic content (TPC) and antioxidant capacity (ABTS •+ ) were determined. Beef burgers with 20 g kg -1 of mortiño (MM) or its PLE extract (ME) were manufactured. Lipid oxidation (TBARS) and instrumental color changes were measured after refrigerated storage. High TPC (up to 72 g gallic acid equivalent kg -1 extract) was determined in mortiño extracts, which was positively correlated with antioxidant activity. TBARS values of beef burgers containing either MM or ME did not change after refrigerated storage, whereas lipid oxidation of control burgers increased significantly. The color of burgers with added MM or ME was different (lower b* and a* values) from that of control burgers. However, the evolution of color after storage was similar between control and ME samples. Mortiño extracts with high TPC can be obtained by PLE. Both mortiño and its PLE extract are able to control lipid oxidation of beef burgers, but the extract is preferred from the color quality point of view. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

A Prototype Combination TPC Cherenkov Detector with GEM Readout for Tracking and Particle Identification and its Potential Use at an Electron Ion Collider

NASA Astrophysics Data System (ADS)

Woody, Craig; Azmoun, Babak; Majka, Richard; Phipps, Michael; Purschke, Martin; Smirnov, Nikolai

2018-02-01

A prototype detector is being developed which combines the functions of a Time Projection Chamber for charged particle tracking and a Cherenkov detector for particle identification. The TPC consists of a 10×10×10 cm3 drift volume where the charge is drifted to a 10×10 cm2 triple GEM detector. The charge is measured on a readout plane consisting of 2×10 mm2 chevron pads which provide a spatial resolution ˜ 100 μm per point in the chevron direction along with dE/dx information. The Cherenkov portion of the detector consists of a second 10×10 cm2 triple GEM with a photosensitive CsI photocathode on the top layer. This detector measures Cherenkov light produced in the drift gas of the TPC by high velocity particles which are above threshold. CF4 or CF4 mixtures will be used as the drift gas which are highly transparent to UV light and can provide excellent efficiency for detecting Cherenkov photons. The drift gas is also used as the operating gas for both GEM detectors. The prototype detector has been constructed and is currently being tested in the lab with sources and cosmic rays, and additional tests are planned in the future to study the detector in a test beam.

Free Radical Scavenging Activity and Comparative Metabolic Profiling of In Vitro Cultured and Field Grown Withania somnifera Roots

PubMed Central

Senthil, Kalaiselvi; Thirugnanasambantham, Pankajavalli; Oh, Taek Joo; Kim, So Hyun; Choi, Hyung Kyoon

2015-01-01

Free radical scavenging activity (FRSA), total phenolic content (TPC), and total flavonoid content (TFC) of in vitro cultured and field grown Withania somnifera (Ashwagandha) roots were investigated. Withanolides analysis and comprehensive metabolic profiling between 100% methanol extracts of in vitro and field grown root tissues was performed using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectrometry (GC-MS), respectively. Significantly higher levels of FRSA, TPC, and TFC were observed in in-vitro cultured roots compared with field grown samples. In addition, 30 day-cultured in vitro root samples (1MIR) exhibited a significantly higher FRSA (IC50 81.01 μg/mL), TPC (118.91 mg GAE/g), and TFC (32.68 mg CE/g) compared with those in 45 day-cultured samples (1.5MIR). Total of 29 metabolites were identified in in vitro cultured and field grown roots by GC-MS analysis. The metabolites included alcohols, organic acids, purine, pyrimidine, sugars, and putrescine. Vanillic acid was only observed in the in vitro cultured root samples, and higher level of the vanillic acid was observed in 1MIR when compared to 1.5MIR. Therefore, it is suggested that 1MIR might serve as an alternative to field grown roots for the development of medicinal and functional food products. PMID:25874568

Aqueous extraction kinetics of soluble solids, phenolics and flavonoids from sage (Salvia fruticosa Miller) leaves.

PubMed

Torun, Mehmet; Dincer, Cuneyt; Topuz, Ayhan; Sahin-Nadeem, Hilal; Ozdemir, Feramuz

2015-05-01

In the present study, aqueous extraction kinetics of total soluble solids (TSS), total phenolic content (TPC) and total flavonoid content (TFC) from Salvia fruticosa leaves were investigated throughout 150 min. of extraction period against temperature (60-80 °C), particle size (2-8 mm) and loading percentage (1-4 %). The extract yielded 25 g/100 g TSS which contained 30 g/100 g TPC and 25 g/100 g TFC. The extraction data in time course fit with reversible first order kinetic model. All tested variables showed significant effect on the estimated kinetic parameters except equilibrium concentration. Increasing the extraction temperature resulted high extraction rate constants and equilibrium concentrations of the tested variables notably above 70 °C. By using the Arrhenius relationship, activation energy of the TSS, TPC and TFC were determined as 46.11 ± 5.61, 36.80 ± 3.12 and 33.52 ± 2.23 kj/mol, respectively. By decreasing the particle size, the extraction rate constants and diffusion coefficients exponentially increased whereas equilibrium concentrations did not change significantly. The equilibrium concentrations of the tested parameters showed linear behavior with increasing the loading percentage of the sage, however; the change in extraction rates did not show linear behavior due to submerging effect of 4 % loading.

Antioxidant activity, total phenolics content, anthocyanin, and color stability of isotonic model beverages colored with Andes berry (Rubus glaucus Benth) anthocyanin powder.

PubMed

Estupiñan, D C; Schwartz, S J; Garzón, G A

2011-01-01

The stability of anthocyanin (ACN) freeze-dried powders from Andes berry (Rubus glaucus Benth) as affected by storage, addition of maltodextrin as a carrier agent, and illumination was evaluated in isotonic model beverages. The ethanolic ACN extract was freeze dried with and without maltodextrin DE 20. Isotonic model beverages were colored with freeze-dried ACN powder (FDA), freeze-dried ACN powder with maltodextrin (MFDA), and red nr 40. Beverages were stored in the dark and under the effect of illumination. Half life of the ACNs, changes in color, total phenolics content (TPC), and antioxidant activity were analyzed for 71 d. Addition of maltodextrin and absence of light stabilized the color of beverages and improved ACN and TPC stability during storage. The antioxidant activity of the beverages was higher when they were colored with MFDA and highly correlated with ACN content. There was no correlation between antioxidant activity and TPC. It is concluded that addition of maltodextrin DE 20 as a carrier agent during freeze-drying improves the color and stability of nutraceutical antioxidants present in Andes berry extract. This suggests a protective enclosing of ACNs within a maltodextrin matrix with a resulting powder that could serve as a supplement or additive to naturally color and to enhance the antioxidant capacity of isotonic beverages.

Synthesis and coordination chemistry of TpC*MI complexes where M=Mg, Ca, Sr, Ba and Zn and TpC*=tris[3-(2-methoxy-1,1-dimethyl)pyrazolyl]hydroborate.

PubMed

Chisholm, Malcolm H; Gallucci, Judith C; Yaman, Gulsah

2009-01-14

Reactions involving MI2 where M=Mg, Ca, Sr, Ba or Zn and M'TpC* where M'=Na or Tl and TpC*=tris[3-methoxy-1,1-dimethyl)pyrazolyl]hydroborate in tetrahydrofuran are described leading to the isolation and characterization of the complexes TpC*MgI, , TpC*CaI, , TpC*SrI, , TpC*SrI(THF), , TpC*BaI, , TpC*BaI(pz*H), , where pz*H=3-(2-methoxyl-1,1-dimethyl)pyrazole, TpC*BaI.1/2toluene, and TpC*ZnI, . The compounds , , , , and have been characterized by single-crystal X-ray crystallography. Compounds and are isostructural and are salt-like containing kappa6-TpM+ cations and I- anions. In all other structures, the iodide is bound to the metal and TpC* is kappa6 bonded to the group 2 M(2+) ions. Reactions involving TpC*CaI, , and sodium or lithium alkoxides or amides failed to yield the amide or alkoxide calcium TpC* derivative, though related reactions involving TpC*ZnI, , and KOSiMe3 proceeded quantitatively to yield kappa3TpC*ZnOSiMe3, , which was also structurally characterized and shown to have the kappa3-TpC* bound ligand.

Characteristics of white, yellow, purple corn accessions: phenolic profile, textural, rheological properties and muffin making potential.

PubMed

Trehan, Shalini; Singh, Narpinder; Kaur, Amritpal

2018-06-01

Physicochemical, antioxidant, phenolic profile and muffin-making properties of white, yellow and purple corn accessions were evaluated. HPLC analysis revealed the presence of ferulic acid, quercetin, sinapic acid, gallic acid and protocatechuic acid in different corn accessions. Ferulic acid was the major phenolic acid present in all accessions. Total phenolic content (TPC), DPPH and ABTS inhibition ranged from 903 to 1843 µg GAE/g, 0.73-0.89 and 3.81-4.92 µM trolox/mg, respectively. Purple accessions had higher TPC than yellow and white accessions. Pasting profiles of different accessions revealed high thermal stability indicated by low breakdown viscosity. Muffin-making properties were determined as batter rheology and muffin specific volume, texture and sensory analysis. Storage modulus ( G ') and loss modulus ( G ″) of batters for white colored exhibited the highest while yellow colored had the lowest value. Firmness and TPC of muffins ranged from 3.1 to 5.9 N and 811-1406 µg GAE/g, respectively. Muffin cohesiveness and chewiness were correlated positively, whereas firmness was negatively related to paste viscosities. Antioxidant activity was correlated to phenolic content of the muffins. Sensory analysis revealed that muffins prepared from yellow corn accession (IC447648) were highly acceptable, while those made from purple corn (IC447644) were not liked much.

Bioactive compounds and antioxidant activities of some cereal milling by-products.

PubMed

Smuda, Sayed Saad; Mohsen, Sobhy Mohamed; Olsen, Karsten; Aly, Mohamed Hassan

2018-03-01

The present study was performed to evaluate the phytochemicals profiles of some cereal milling by-products such as wheat (bran, germ and shorts), rice (bran, germ and husk) and corn (bran, germ and germ meal) to assess their potentiality as bioactive compounds sources. Distilled water, ethanol, methanol, and acetone separately were used as solvents for the extraction of phytochemicals compounds. The antioxidant activity (AOA), total phenolics content (TPC), and total flavonoids content (TFC) of the extracts were investigated using various in vitro assays. The results showed that tannins content was ranged from 113.4 to 389.5 (mg/100 g sample).The study revealed that TPC and TFC of cereal by-products extracts were significantly different for various solvents. TPC content varied from 366.1 to 1924.9 mg/100 g and TFC content varied from 139.3 to 681.6 mg/100 g. High carotenoids content was observed for corn germ meal and minimum for wheat bran. Distilled water, ethanol and methanol extracts showed significantly different antioxidant activity. Significant variations were observed with regard to AOA of different cereal by-products by using various solvents. The ethanol and methanol were observed to be the best solvents to extract phenolic compounds and antioxidant activity, while acetone extract showed less efficiency. Also, the cereal milling by-products were rich in bioactive compounds and could be used as a value added products.

Expression and identification of 10 sarcomeric MyHC isoforms in human skeletal muscles of different embryological origin. Diversity and similarity in mammalian species.

PubMed

Mascarello, Francesco; Toniolo, Luana; Cancellara, Pasqua; Reggiani, Carlo; Maccatrozzo, Lisa

2016-09-01

In the mammalian genome, among myosin heavy chain (MyHC) isoforms a family can be identified as sarcomeric based on their molecular structure which allows thick filament formation. In this study we aimed to assess the expression of the 10 sarcomeric isoforms in human skeletal muscles, adopting this species as a reference for comparison with all other mammalian species. To this aim, we set up the condition for quantitative Real Time PCR assay to detect and quantify MyHC mRNA expression in a wide variety of human muscles from somitic, presomitic and preotic origin. Specific patterns of expression of the following genes MYH1, MYH2, MYH3, MYH4, MYH6, MYH7, MYH8, MYH13, MYH14/7b and MYH15 were demonstrated in various muscle samples. On the same muscle samples which were analysed for mRNA expression, the corresponding MyHC proteins were studied with SDS PAGE and Western blot. The mRNA-protein comparison allowed the identification of 10 distinct proteins based on the electrophoretic migration rate. Three groups were formed based on the migration rate: fast migrating comprising beta/slow/1, alpha cardiac and fast 2B, slow migrating comprising fast 2X, fast 2A and two developmental isoforms (NEO and EMB), intermediate migrating comprising EO MyHC, slow B (product of MYH15), slow tonic (product of MYH14/7b). Of special interest was the demonstration of a protein band corresponding to 2B-MyHC in laryngeal muscles and the finding that all 10 isoforms are expressed in extraocular muscles. These latter muscles are the unique localization for extraocular, slow B (product of MYH15) and slow tonic (product of MYH14/7b). Copyright © 2016 Elsevier GmbH. All rights reserved.

Expression of membrane-bound and cytosolic guanylyl cyclases in the rat inner ear.

PubMed

Seebacher, T; Beitz, E; Kumagami, H; Wild, K; Ruppersberg, J P; Schultz, J E

1999-01-01

Membrane-bound guanylyl cyclases (GCs) are peptide hormone receptors whereas the cytosolic isoforms are receptors for nitric oxide. In the inner ear, the membrane-bound GCs may be involved in the regulation of fluid homeostasis and the cytosolic forms possibly play a role in signal processing and regulation of local blood flow. In this comprehensive study, we examined, qualitatively and quantitatively, the transcription pattern of all known GC isoforms in the inner ear from rat by RT-PCR. The tissues used were endolymphatic sac, stria vascularis, organ of Corti, organ of Corti outer hair cells, cochlear nerve, Reissner's membrane, vestibular dark cells, and vestibular sensory cells. We show that multiple particulate (GC-A, GC-B, GC-D, GC-E, GC-F and GC-G) and several subunits of the heterodimeric cytosolic GCs (alpha1, alpha2, beta1 and beta2) are expressed, albeit at highly different levels. GC-C was not found. GC-A and the soluble subunits alpha1 and beta1 were transcribed ubiquitously. GC-B was present in all tissues except stria vascularis, which contained GC-A and traces of GC-E and GC-G. GC-B was by far the predominant membrane-bound isoform in the organ of Corti (86%), Reissner's membrane (75%) and the vestibulum (80%). Surprisingly, GC-E, a retinal isoform, was detected in significant amounts in the cochlear nerve (8%) and in the organ of Corti (4%). Although the cytosolic GC is a heterodimer composed of an alpha and a beta subunit, the mRNA transcription of these subunits was not stoichiometric. Particularly in the vestibulum, the transcription of the beta1 subunits was at least four-fold higher than of the alpha1 subunit. The data are compatible with earlier suggestions that membrane receptor GCs may be involved in the control of inner ear electrolyte and fluid composition whereas NO-stimulated GC isoforms mainly participate in the regulation of blood flow and supporting cell physiology.

Osteoblast gene expression is differentially regulated by TGF-beta isoforms.

PubMed

Fagenholz, P J; Warren, S M; Greenwald, J A; Bouletreau, P J; Spector, J A; Crisera, F E; Longaker, M T

2001-03-01

The transforming growth factor beta (TGF-beta) superfamily encompasses a number of important growth factors including several TGF-beta isoforms, the bone morphogenetic proteins, activins, inhibins, and growth and differentiation factors. TGF-beta 1, -beta 2, and -beta 3 are three closely related isoforms that are widely expressed during skeletal morphogenesis and bone repair. Numerous studies suggest that each isoform has unique in vivo functions; however, the effects of these TGF-beta isoforms on osteoblast gene expression and maturation have never been directly compared. In the current study, we treated undifferentiated neonatal rat calvaria osteoblast-enriched cell cultures with 2.5 ng/ml of each TGF-beta isoform and analyzed gene expression at 0, 3, 6, and 24 hours. We demonstrated unique isoform-specific regulation of endogenous TGF-beta 1 and type I collagen mRNA transcription. To assess the effects of extended TGF-beta treatment on osteoblast maturation, we differentiated osteoblast cultures in the presence of 2.5 ng/ml of each TGF-beta isoform. Analysis of collagen I, alkaline phosphatase, and osteocalcin demonstrated that each TGF-beta isoform uniquely suppressed the transcription of these osteoblast differentiation markers. Interestingly, TGF-beta isoform treatment increased osteopontin expression in primary osteoblasts after 4 and 10 days of differentiation. To our knowledge, these data provide the first direct comparison of the effects of the TGF-beta isoforms on osteoblast gene expression in vitro. Furthermore, these data suggest that TGF-beta isoforms may exert their unique in vivo effects by differentially regulating osteoblast cytokine secretion, extracellular matrix production, and the rate of cellular maturation.

Crystal structures of a halophilic archaeal malate synthase from Haloferax volcanii and comparisons with isoforms A and G

PubMed Central

2011-01-01

Background Malate synthase, one of the two enzymes unique to the glyoxylate cycle, is found in all three domains of life, and is crucial to the utilization of two-carbon compounds for net biosynthetic pathways such as gluconeogenesis. In addition to the main isoforms A and G, so named because of their differential expression in E. coli grown on either acetate or glycolate respectively, a third distinct isoform has been identified. These three isoforms differ considerably in size and sequence conservation. The A isoform (MSA) comprises ~530 residues, the G isoform (MSG) is ~730 residues, and this third isoform (MSH-halophilic) is ~430 residues in length. Both isoforms A and G have been structurally characterized in detail, but no structures have been reported for the H isoform which has been found thus far only in members of the halophilic Archaea. Results We have solved the structure of a malate synthase H (MSH) isoform member from Haloferax volcanii in complex with glyoxylate at 2.51 Å resolution, and also as a ternary complex with acetyl-coenzyme A and pyruvate at 1.95 Å. Like the A and G isoforms, MSH is based on a β8/α8 (TIM) barrel. Unlike previously solved malate synthase structures which are all monomeric, this enzyme is found in the native state as a trimer/hexamer equilibrium. Compared to isoforms A and G, MSH displays deletion of an N-terminal domain and a smaller deletion at the C-terminus. The MSH active site is closely superimposable with those of MSA and MSG, with the ternary complex indicating a nucleophilic attack on pyruvate by the enolate intermediate of acetyl-coenzyme A. Conclusions The reported structures of MSH from Haloferax volcanii allow a detailed analysis and comparison with previously solved structures of isoforms A and G. These structural comparisons provide insight into evolutionary relationships among these isoforms, and also indicate that despite the size and sequence variation, and the truncated C-terminal domain of the H isoform, the catalytic mechanism is conserved. Sequence analysis in light of the structure indicates that additional members of isoform H likely exist in the databases but have been misannotated. PMID:21569248

Hormonal regulation of circulating insulin-like growth factor-binding protein-1 phosphorylation status.

PubMed

Westwood, M; Gibson, J M; Williams, A C; Clayton, P E; Hamberg, O; Flyvbjerg, A; White, A

1995-12-01

Insulin-like growth factor (IGF)-binding protein-1 (IGFBP-1) normally circulates as a single, highly phosphorylated species. However, IGFBP-1 phosphorylation status can be altered, such as in pregnancy where non- and lesser phosphorylated isoforms are also present. We have examined how hormonal regulators of circulating IGFBP-1 influence its phosphorylation status and, hence, its ability to modulate IGF activity. In response to insulin-induced hypoglycemia (0.2 U/kg, iv), an increase in the highly phosphorylated isoform was observed after 5 h [16 (range, 11.5-35.5) to 77 (range, 63-250) microgram/L; 4.8-fold increase; P = 0.009], but no non- or lesser phosphorylated variants could be detected. Glucagon (1 mg, sc), increased IGFBP-1 from 27 (range, 13-36.5) to 112 (range, 100.5-129) micrograms/L (4.1-fold increase; P = 0.009) after 90 min despite preceding insulin concentrations of more than 500 pmol/L, but again the IGFBP-1 remained in the highly phosphorylated form. Regulation of IGFBP-1 phosphorylation by sex steroids was studied by comparing women receiving a combined oral contraceptive with women on no medication. Although plasma IGFBP-1 levels were significantly elevated in the treatment group [120 (range, 97.5-237.5) vs. 52 (range, 38-70) micrograms/L; P < 0.004], there was no difference in the form of IGFBP-1 present. The acute effect of somatostatin (500 micrograms/h) on IGFBP-1 phosphorylation status was also studied. Somatostatin only increased the phosphoform characteristic of normal subjects; the appearance of non- or lesser phosphorylated variants was not induced. The effect of rhIGF-I (80 or 120 micrograms, sc) on plasma IGFBP-1 was studied in three subjects with Laron's syndrome. A transient increase in the highly phosphorylated isoform of IGFBP-1 was noted; there was no rise in the non- and lesser phosphorylated isoforms also found in the plasma of Laron's syndrome subjects. These data suggest that only the highly phosphorylated species of IGFBP-1 is under hormonal control; regulation of the non- and lesser phosphorylated variants remains to be determined.