CFP-10 from Mycobacterium tuberculosis Selectively Activates Human Neutrophils through a Pertussis Toxin-Sensitive Chemotactic Receptor

PubMed Central

Björnsdottir, Halla; Winther, Malene; Christenson, Karin; Oprea, Tudor; Karlsson, Anna; Forsman, Huamei; Dahlgren, Claes; Bylund, Johan

2014-01-01

Upon infection with Mycobacterium tuberculosis, neutrophils are massively recruited to the lungs, but the role of these cells in combating the infection is poorly understood. Through a type VII secretion system, M. tuberculosis releases a heterodimeric protein complex, containing a 6-kDa early secreted antigenic target (ESAT-6) and a 10-kDa culture filtrate protein (CFP-10), that is essential for virulence. Whereas the ESAT-6 component possesses multiple virulence-related activities, no direct biological activity of CFP-10 has been shown, and CFP-10 has been described as a chaperone protein for ESAT-6. We here show that the ESAT-6:CFP-10 complex induces a transient release of Ca2+ from intracellular stores in human neutrophils. Surprisingly, CFP-10 rather than ESAT-6 was responsible for triggering the Ca2+ response, in a pertussis toxin-sensitive manner, suggesting the involvement of a G-protein-coupled receptor. In line with this, the response was accompanied by neutrophil chemotaxis and activation of the superoxide-producing NADPH-oxidase. Neutrophils were unique among leukocytes in responding to CFP-10, as monocytes and lymphocytes failed to produce a Ca2+ signal upon stimulation with the M. tuberculosis protein. Hence, CFP-10 may contribute specifically to neutrophil recruitment and activation during M. tuberculosis infection, representing a novel biological role for CFP-10 in the ESAT-6:CFP-10 complex, beyond the previously described chaperone function. PMID:25332123

Ciguatera fish poisoning and environmental change: a case for strengthening health surveillance in the Pacific?

PubMed

Derne, Bonnie; Fearnley, Emily; Goater, Sarah; Carter, Karen; Weinstein, Philip

2010-09-01

Ciguatera Fish Poisoning (CFP), a significant public health problem in the Pacific, is intrinsically linked to the health of coral reef ecosystems. Incidence data on CFP could therefore be used, in theory, as indicators of disruption to coral reefs. Some disruptions, such as increasing sea surface temperatures, result from global environmental change--therefore suggesting that CFP is likely to become an increasing public health problem in the region. The proactive management of increasing numbers of cases will depend on an understanding of the ecology of the disease, sound health surveillance systems to report cases of CFP including appropriate case definitions, and quantifiable correlations between case numbers and environmental variables. Here, we briefly review the knowledge about these components in Pacific Island Countries and Territories (PICTs), including summarising regional variation in symptoms of CFP cases, investigating media as an enhanced surveillance tool, and summarising regional environmental drivers of CFP cases. We conclude that CFP could be an important indicator of the health of reef ecosystems in the face of global climate change and more novel approaches such as combining environmental and health data, need to be implemented to improve surveillance of CFP.

A review of traditional remedies of ciguatera fish poisoning in the Pacific.

PubMed

Kumar-Roiné, Shilpa; Taiana Darius, H; Matsui, Mariko; Fabre, Nicolas; Haddad, Mohamed; Chinain, Mireille; Pauillac, Serge; Laurent, Dominique

2011-07-01

Ciguatera fish poisoning (CFP) is an illness caused by eating tropical coral fish contaminated with ciguatoxins (CTXs). The clinical management of patients with CFP is generally supportive and symptomatic in nature as no antidote exists. Of the many drugs prescribed, several have been claimed to be efficient in small, uncontrolled studies, but the outcomes of treatments with these medicines are often contradictory. In New Caledonia, traditional remedies are commonly employed in the treatment of CFP and of the 90 plant species catalogued as useful in CFP, the most popular herbal remedy by far is a decoction prepared from the leaves of Heliotropium foertherianum Diane & Hilger (Boraginaceae). Other important plants used in the treatment of CFP include Euphorbia hirta L. (Euphorbiaceae) and Vitex L. sp. (Lamiaceae). This review focuses on the evidence for efficacy of these species and pharmacological studies which support their use. Other plants used in CFP and the conventional treatment of CFP are also discussed briefly. Copyright © 2011 John Wiley & Sons, Ltd.

Roles of NMDA and dopamine D1 and D2 receptors in the acquisition and expression of flavor preferences conditioned by oral glucose in rats.

PubMed

Dela Cruz, J A D; Coke, T; Icaza-Cukali, D; Khalifa, N; Bodnar, R J

2014-10-01

Animals learn to prefer flavors associated with the intake of sugar (sucrose, fructose, glucose) and fat (corn oil: CO) solutions. Conditioned flavor preferences (CFP) have been elicited for sugars based on orosensory (flavor-flavor: e.g., fructose-CFP) and post-ingestive (flavor-nutrient: e.g., intragastric (IG) glucose-CFP) processes. Dopamine (DA) D1, DA D2 and NMDA receptor antagonism differentially eliminate the acquisition and expression of fructose-CFP and IG glucose-CFP. However, pharmacological analysis of fat (CO)-CFP, mediated by both flavor-flavor and flavor-nutrient processes, indicated that acquisition and expression of fat-CFP were minimally affected by systemic DA D1 and D2 antagonists, and were reduced by NMDA antagonism. Therefore, the present study examined whether systemic DA D1 (SCH23390), DA D2 (raclopride) or NMDA (MK-801) receptor antagonists altered acquisition and/or expression of CFP induced by oral glucose that should be mediated by both flavor-flavor and flavor-nutrient processes. Oral glucose-CFP was elicited following by training rats to drink one novel flavor (CS+, e.g., cherry) mixed in 8% glucose and another flavor (CS-, e.g., grape) mixed in 2% glucose. In expression studies, food-restricted rats drank these solutions in one-bottle sessions (2 h) over 10 days. Subsequent two-bottle tests with the CS+ and CS- flavors mixed in 2% glucose occurred 0.5 h after systemic administration of vehicle (VEH), SCH23390 (50-800 nmol/kg), raclopride (50-800 nmol/kg) or MK-801 (50-200 μg/kg). Rats displayed a robust CS+ preference following VEH treatment (94-95%) which was significantly though marginally attenuated by SCH23390 (67-70%), raclopride (77%) or MK-801 (70%) at doses that also markedly reduced overall CS intake. In separate acquisition studies, rats received VEH, SCH23390 (50-400 nmol/kg), raclopride (50-400 nmol/kg) or MK-801 (100 μg/kg) 0.5 h prior to ten 1-bottle training trials with CS+/8%G and CS-/2%G training solutions that was followed by six 2-bottle CS+ vs. CS- tests in 2% glucose conducted without injections. The significant and persistent CS+ preferences observed in the VEH (94-98%) group was significantly reduced by rats receiving SCH23390 at 400 nmol/kg (65-73%), raclopride at 200 or 400 nmol/kg (76-82%) or MK-801 at 100 μg/kg (68-69%). Thus, systemic DA D1 and DA D2 receptor antagonism produced smaller reductions in the expression of oral glucose-CFP relative to fructose-CFP or IG-glucose-CFP. Correspondingly, systemic DA D1, DA D2 and NMDA receptor antagonism also produced smaller reductions in the acquisition of oral glucose-CFP relative to fructose-CFP or IG-glucose-CFP. These data suggest, but do not prove, that the magnitude and persistence of these receptor antagonist effects upon sugar-CFP might depend upon the individual or combined engagement of flavor-flavor and flavor-nutrient processes. Copyright © 2014 Elsevier Inc. All rights reserved.

Vertically aligned carbon nanotubes/carbon fiber paper composite to support Pt nanoparticles for direct methanol fuel cell application

NASA Astrophysics Data System (ADS)

Zhang, Jing; Yi, Xi-bin; Liu, Shuo; Fan, Hui-Li; Ju, Wei; Wang, Qi-Chun; Ma, Jie

2017-03-01

Vertically aligned carbon nanotubes (VACNTs) grown on carbon fiber paper (CFP) by plasma enhanced chemical vapor deposition is introduced as a catalyst support material for direct methanol fuel cells (DMFCs). Well dispersed Pt nanoparticles on VACNTs surface are prepared by impregnation-reduction method. The VACNTs on CFP possess well-maintained alignment, large surface area and good electrical conductivity, which leading to the formation of Pt particles with a smaller size and enhance the Pt utilization rate. The structure and nature of resulting Pt/VACNTs/CFP catalysts for methanol oxidation are investigated by transmission electron microscopy (TEM), X-ray diffraction (XRD) and scanning electron microscope (SEM). With the aid of VACNTs, well-dispersed Pt catalysts enable the reversibly rapid redox kinetic since electron transport efficiently passes through a one-dimensional pathway, which leads to enhance the catalytic activity and Pt utilization rate. Compared with the Pt/XC-72/CFP electrode, the electrochemical measurements results display that the Pt/VACNTs/CFP catalyst shows much higher electrocatalytic activity and better stability for methanol oxidation. In addition, the oxidation current from 200 to 1200 s decayed more slowly for the Pt/VACNTs/CFP than that of the Pt/XC-72/CFP catalysts, indicating less accumulation of adsorbed CO species. All those results imply that the Pt/VACNTs/CFP has a great potential for applications in DMFCs.

Thin layer chitosan-coated cellulose filter paper as substrate for immobilization of catalytic cobalt nanoparticles.

PubMed

Kamal, Tahseen; Khan, Sher Bahadar; Haider, Sajjad; Alghamdi, Yousef Gamaan; Asiri, Abdullah M

2017-11-01

A facile approach utilizing synthesis of cobalt nanoparticles in green polymers of chitosan (CS) coating layer on high surface area cellulose microfibers of filter paper (CFP) is described for the catalytic reduction of nitrophenol and an organic dye using NaBH 4 . Simple steps of CFP coating with 1wt% CS aqueous solution followed by Co 2+ ions adsorption from 0.2M CoCl 2 aqueous solution were carried out to prepare pre-catalytic strips. The Co 2+ loaded pre-catalytic strips of CS-CFP were treated with 0.19M NaBH 4 aqueous solution to convert the ions into nanoparticles. Successful Co nanoparticles formation was assessed by various characterization techniques of FESEM, EDX and XRD analyzes. TGA analyses were carried out on CFP, CS-CFP, and Co-CS-CFP for the determination of the amount of Co particles formed on the CS-FP, and to track their thermal properties. Furthermore, we demonstrated that the Co-CS-CFP showed an excellent catalytic activity and reusability in the reduction reactions a nitroaromatic compound of 2,6-dintirophenol (2,6-DNP) and brilliant cresyl blue (BCB) dye by NaBH 4 . The Co-CS-CFP catalyzed the reduction reactions of 2,6-DNP and BCB by NaBH 4 with psuedo-first order rate constants of 0.0451 and 0.1987min -1 , respectively. Copyright © 2017 Elsevier B.V. All rights reserved.

Comparison of conventional color fundus photography and multicolor imaging in choroidal or retinal lesions.

PubMed

Muftuoglu, Ilkay Kilic; Gaber, Raouf; Bartsch, Dirk-Uwe; Meshi, Amit; Goldbaum, Michael; Freeman, William R

2018-04-01

Our purpose was to compare the characteristics of the retinal and choroidal lesions including choroidal nevus, choroidal melanoma and congenital hypertrophy of the retina pigment epithelium using conventional color fundus photography (CFP) and multicolor imaging (MCI). The paired images of patients with retinal or choroidal lesions were assessed for the visibility of lesion's border, halo and drusen using a grading scale (0-2). The area of the lesion was measured on both imaging modalities. The same grading was also done on the individual color channels of MCI for a further evaluation. Thirty-three eyes of 33 patients were included. There were no significant differences in the mean border, drusen and halo visibility scores between the two imaging modalities (p = 0.12, p = 0.70, p = 0.35). However, the mean area of the lesion was significantly smaller on MCI than that on CFP (14.9±3.3 versus 18.7±3.4 mm 2 , p = 0.01). The appearance of choroidal and/ or retinal lesions on MCI may be different than that on CFP. Though MCI can provide similar information with CFP for the features of retinal and/ or choroidal lesions including border, halo and drusen; the infrared light reflection on MCI underestimates the extent of the choroidal lesion by 33%.

[Expression of the fusion protein CFP10-ESAT6 of Mycobacterium tuberculosis and the study of its immunogenicity].

PubMed

Wang, Xiao-ying; Bao, Lang; Zhao, Ming-cai; Zhang, Hui-dong; Long, Yang

2006-05-01

To express a recombinant fusion protein CFP10-ESAT6 of Mycobacterium tuberculosis, and obtain the polyclonal antibodies of this fusion protein by immune rabbit. The 630 bp cfpl0-esat6 fusion gene fragments were amplified from the genomic DNA of a Mycobacterium tuberculosis reference strain H37Rv and inserted into the expression plasmid pET32a (+) to generate the recombinant plasmid pET-cfp10-esat6. The recombinat expression plasmid was transformed into E. coli BL21 (DE3). The fused protein CFP10-ESAT6 with His-tag was expressed after inducing with IPTG and purified with affinity chromatography. This protein was used to immune the rabbit to obtained the polyclonal antibodies, and been analyzed with Western-blot and ELISA. The recombinant plasmid pET-cfp10-esat6 was success fully constructed, the recombinant fusion protein CFP10-ESAT6 could be expressed at relatively high levels, and the polyclonal antibodies of fusion protein were obtained. The successful construction and expression of the recombinant fusion protein CFP10-ESAT6 and the obtained polyclonal antibodies will be very helpful for the development of new anti-tuberculosis vaccine and the clinical serologic diagnosis.

Quasi-static and dynamic motions of the columellar footplate in ostrich (Struthio camelus) measured ex vivo.

PubMed

Muyshondt, Pieter G G; Claes, Raf; Aerts, Peter; Dirckx, Joris J J

2018-01-01

The nature of the movement of the columellar footplate (CFP) in birds is still a matter of ongoing debate. Some sources claim that rocking motion is dominant, while others propose a largely piston-like motion. In this study, motions of the CFP are experimentally investigated in the ostrich using a post-mortem approach. For quasi-static loads, micro-CT scans of ostrich heads were made under positive and negative middle-ear pressures of 1 kPa. For dynamic loads, laser Doppler vibrometry was used to measure the velocity on multiple locations of the CFP as a function of excitation frequency from 0.125 to 4 kHz, and digital stroboscopic holography was used to assess the 1D full-field out-of-plane displacement of the CFP at different excitation frequencies. To expose the CFP in the experiments, measurements were made from the medial side of the CFP after opening and draining the inner ear. To determine the influence of the inner-ear load on CFP motions, a finite element model was created of the intact ostrich middle ear with inner-ear load included. For quasi-static loads, the CFP performed largely piston-like motions under positive ME pressure, while under negative ME pressure the difference between piston and rocking motion was smaller. For dynamic loads, the CFP motion was almost completely piston-like for frequencies below 1 kHz. For higher frequencies, the motions became more complicated with an increase of the rocking components, although they never exceeded the piston component. When including the inner-ear load to the model, the rocking components started to increase relative to the piston component when compared to the result of the model with unloaded CFP, but only at high frequencies above 1 kHz. In this frequency range, the motion could no longer be identified as purely piston-like or rocking. As a conclusion, the current results suggest that CFP motion is predominantly piston-like below 1 kHz, while at higher frequencies the motion becomes too complicated to be described as purely piston-like or rocking. Copyright © 2017 Elsevier B.V. All rights reserved.

Update on methodologies available for ciguatoxin determination: perspectives to confront the onset of ciguatera fish poisoning in Europe.

PubMed

Caillaud, Amandine; de la Iglesia, Pablo; Darius, H Taiana; Pauillac, Serge; Aligizaki, Katerina; Fraga, Santiago; Chinain, Mireille; Diogène, Jorge

2010-06-14

Ciguatera fish poisoning (CFP) occurs mainly when humans ingest finfish contaminated with ciguatoxins (CTXs). The complexity and variability of such toxins have made it difficult to develop reliable methods to routinely monitor CFP with specificity and sensitivity. This review aims to describe the methodologies available for CTX detection, including those based on the toxicological, biochemical, chemical, and pharmaceutical properties of CTXs. Selecting any of these methodological approaches for routine monitoring of ciguatera may be dependent upon the applicability of the method. However, identifying a reference validation method for CTXs is a critical and urgent issue, and is dependent upon the availability of certified CTX standards and the coordinated action of laboratories. Reports of CFP cases in European hospitals have been described in several countries, and are mostly due to travel to CFP endemic areas. Additionally, the recent detection of the CTX-producing tropical genus Gambierdiscus in the eastern Atlantic Ocean of the northern hemisphere and in the Mediterranean Sea, as well as the confirmation of CFP in the Canary Islands and possibly in Madeira, constitute other reasons to study the onset of CFP in Europe [1]. The question of the possible contribution of climate change to the distribution of toxin-producing microalgae and ciguateric fish is raised. The impact of ciguatera onset on European Union (EU) policies will be discussed with respect to EU regulations on marine toxins in seafood. Critical analysis and availability of methodologies for CTX determination is required for a rapid response to suspected CFP cases and to conduct sound CFP risk analysis.

Update on Methodologies Available for Ciguatoxin Determination: Perspectives to Confront the Onset of Ciguatera Fish Poisoning in Europe [1

PubMed Central

Caillaud, Amandine; de la Iglesia, Pablo; Darius, H. Taiana; Pauillac, Serge; Aligizaki, Katerina; Fraga, Santiago; Chinain, Mireille; Diogène, Jorge

2010-01-01

Ciguatera fish poisoning (CFP) occurs mainly when humans ingest finfish contaminated with ciguatoxins (CTXs). The complexity and variability of such toxins have made it difficult to develop reliable methods to routinely monitor CFP with specificity and sensitivity. This review aims to describe the methodologies available for CTX detection, including those based on the toxicological, biochemical, chemical, and pharmaceutical properties of CTXs. Selecting any of these methodological approaches for routine monitoring of ciguatera may be dependent upon the applicability of the method. However, identifying a reference validation method for CTXs is a critical and urgent issue, and is dependent upon the availability of certified CTX standards and the coordinated action of laboratories. Reports of CFP cases in European hospitals have been described in several countries, and are mostly due to travel to CFP endemic areas. Additionally, the recent detection of the CTX-producing tropical genus Gambierdiscus in the eastern Atlantic Ocean of the northern hemisphere and in the Mediterranean Sea, as well as the confirmation of CFP in the Canary Islands and possibly in Madeira, constitute other reasons to study the onset of CFP in Europe [1]. The question of the possible contribution of climate change to the distribution of toxin-producing microalgae and ciguateric fish is raised. The impact of ciguatera onset on European Union (EU) policies will be discussed with respect to EU regulations on marine toxins in seafood. Critical analysis and availability of methodologies for CTX determination is required for a rapid response to suspected CFP cases and to conduct sound CFP risk analysis. PMID:20631873

Optimization of Computerized Drills: An Instructional Approach.

ERIC Educational Resources Information Center

Siegel, Martin A.; DiBello, Louis V.

This report describes a computer drill called the Corrective Feedback Paradigm (CFP) and summarizes a research study which examines the effects of the CFP on posttest and time to mastery achievement. The CFP, which embodies the use of such principles as mastery learning, increasing ratio review, and discrimination training, is capable of being…

Improving the conversion of biomass in catalytic fast pyrolysis via white-rot fungal pretreatment.

PubMed

Yu, Yanqing; Zeng, Yelin; Zuo, Jiane; Ma, Fuying; Yang, Xuewei; Zhang, Xiaoyu; Wang, Yujue

2013-04-01

This study investigated the effect of white-rot fungal pretreatment on corn stover conversion in catalytic fast pyrolysis (CFP). Corn stover pretreated by white-rot fungus Irpex lacteus CD2 was fast pyrolyzed alone (non-CFP) and with ZSM-5 zeolite (CFP) in a semi-batch pyroprobe reactor. The fungal pretreatment considerably increased the volatile product yields (predominantly oxygenated compounds) in non-CFP, indicating that fungal pretreatment enhances the corn stover conversion in fast pyrolysis. In the presence of ZSM-5 zeolite, these oxygenated volatiles were further catalytically converted to aromatic hydrocarbons, whose yield increased from 10.03 wt.% for the untreated corn stover to 11.49 wt.% for the pretreated sample. In contrast, the coke yield decreased from 14.29 to 11.93 wt.% in CFP following the fungal pretreatment. These results indicate that fungal pretreatment can enhance the production of valuable aromatics and decrease the amount of undesired coke, and thus has a beneficial effect on biomass conversion in CFP. Copyright © 2013 Elsevier Ltd. All rights reserved.

Role of angiotensin converting enzyme in the vascular effects of an endopeptidase 24.15 inhibitor.

PubMed Central

Telford, S E; Smith, A I; Lew, R A; Perich, R B; Madden, A C; Evans, R G

1995-01-01

1. We investigated the role of angiotensin converting enzyme (ACE) in the cardiovascular effects of N-[1-(R,S)-carboxy-3-phenylpropyl]-Ala-Ala-Tyr-p-aminobenzoate (cFP), a peptidase inhibitor selective for metalloendopeptidase (EP) E.C. 3.4.24.15. 2. In conscious rabbits, cFP (5 mg kg-1, i.v.) markedly slowed the degradation of [3H]-bradykinin, potentiated the depressor response to right atrial administration of bradykinin (10-1000 ng kg-1), and inhibited the pressor response to right atrial angiotensin I (10-100 ng kg-1). In each of these respects, the effects of cFP were indistinguishable from those of the ACE inhibitor, captopril (0.5 mg plus 10 mg kg-1h-1 i.v.). Furthermore, the effects of combined administration of cFP and captopril were indistinguishable from those of captopril alone. 3. In experimentally naive anaesthetized rats, cFP administration (9.3 mg kg-1, i.v.) was followed by a moderate but sustained fall in arterial pressure of 13 mmHg. However, in rats pretreated with bradykinin (50 micrograms kg-1) a more pronounced fall of 30 mmHg was observed. Captopril (5 mg kg-1) had similar hypotensive effects to those of cFP, and cFP had no effect when it was administered after captopril. 4. CFP displaced the binding of [125I]-351A (the p-hydroxybenzamidine derivative of lisinopril) from preparations of rat plasma ACE and solubilized lung membrane ACE (KD = 1.2 and 0.14 microM respectively), and inhibited rat plasma ACE activity (KI = 2.4 microM). Addition of phosphoramidon (10 microM), an inhibitor of a range of metalloendopeptidases, including neutral endopeptidase (E.C.3.4.24.11), markedly reduced the potency of cFP in these systems.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7620708

Toward High-Performance and Low-Cost Hydrogen Evolution Reaction Electrocatalysts: Nanostructuring Cobalt Phosphide (CoP) Particles on Carbon Fiber Paper.

PubMed

Yu, Shu Hearn; Chua, Daniel H C

2018-05-02

In this communication, we facily fabricated nanostructured CoP particles (150 to 200 nm) on carbon fiber paper (CFP) for hydrogen evolution reaction (HER) by a simple two-step process via a green route. In the first step, crystalline Co 3 O 4 nanocubes (150-200 nm) were loaded on CFP through a hydrothermal process at low temperature (120 °C). Interestingly, crystalline Co 3 O 4 nanocubes with a size 150-200 nm exhibited different growth mechanisms in contrast to the crystalline Co 3 O 4 nanocubes with a size <100 nm reported earlier. In the second step, these crystalline Co 3 O 4 nanocubes were converted to catalytically active CoP particles through chemical vapor deposition (CVD) phosphorization (denoted as CoP/CFP-H). Remarkably, CoP/CFP-H exhibited a low Tafel slope of 49.7 mV/dec and only required overpotentials of 128.1, 144.4, and 190.8 mV to drive geometric current densities of -10, -20, and -100 mA cm -2 , respectively. Besides, the CoP/CFP-H also demonstrated an excellent durability in an acidic environment under 2000 sweeps at a high scan rate (100 mV s -1 ) and a 24 h chronopotentiometry testing. For comparison, CoP was also fabricated through the electrodeposition method, followed by CVD phosphorization (denoted as CoP/CFP-E). It was found that the latter had exhibited inferior activity compared to CoP/CFP-H. The good performances of CoP/CFP-H are essentially due to the rational designs of electrode: (i) the applications of highly HER active CoP electrocatalyst, (ii) the intimate contact of nanostructured CoP on carbon fibers, and (iii) the large electrochemical surface area at electrocatalyst/electrolyte interface due to the large retaining of particles features after phosphorization. Notably, the intermediate Co 3 O 4 /CFP can serve as a platform to develop other cobalt-based functional materials.

The role of genetics in fisheries management under the E.U. common fisheries policy.

PubMed

Casey, J; Jardim, E; Martinsohn, J Th

2016-12-01

Exploitation of fish and shellfish stocks by the European Union fishing fleet is managed under the Common Fisheries Policy (CFP), which aims to ensure that fishing and aquaculture are environmentally, economically and socially sustainable and that they provide a source of healthy food for E.U. citizens. A notable feature of the CFP is its legally enshrined requirement for sound scientific advice to underpin its objectives. The CFP was first conceived in 1970 when it formed part of the Common Agricultural Policy. Its formal inception as a stand-alone regulation occurred in 1983 and since that time, the CFP has undergone reforms in 1992, 2002 and 2013, each time bringing additional challenges to the scientific advisory process as the scope of the advice increased in response to changing objectives arising from E.U. regulations and commitments to international agreements. This paper reviews the influence that genetics has had on fish stock assessments and the provision of management advice for European fisheries under successive reforms of the CFP. The developments in genetics since the inception of the CFP have given rise to a diverse and versatile set of genetic techniques that have the potential to provide significant added value to fisheries assessments and the scientific advisory process. While in some cases, notably Pacific salmon Oncorhynchus spp., genetics appear to be very well integrated into existing management schemes, it seems that for marine fishes, discussions on the use of genetics and genomics for fisheries management are often driven by the remarkable technological progress in this field, rather than imminent needs emerging from policy frameworks. An example is the recent suggestion to use environmental (e)DNA for monitoring purposes. While there is no denying that state-of-the-art genetic and genomic approaches can and will be of value to address a number of issues relevant for the management and conservation of marine renewable natural resources, a focus on technology rather than policy and management needs is prone to widen the gap between science and policy, governance and management, thereby further impeding the effective integration of genetic and genomic information into the fisheries management decision making process. Hence, rather than focusing on what is technically achievable, this review outlines suggestions as to which modern genetic and genomic approaches are likely to help address some of the most pressing fisheries management challenges under the CFP. © 2016 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of The Fisheries Society of the British Isles.

Unexpected rare complication of the facial paralysis in a patient with an antrochoanal polyp following canine fossa puncture.

PubMed

Kim, Ah-Young; Choi, Myoung Su

2015-05-14

Canine fossa puncture (CFP) combined with endoscopic sinus surgery is a simple and effective method for treating antrochoanal polyps, particularly those that originate in the anterior, inferior or medial aspect of the antrum. Several complications can occur following CFP, including facial paraesthesia and dental numbness. However, facial palsy is extremely rare after CFP. We postulated that a possible mechanism of facial palsy is pressure injury to the soft tissues adjacent to the puncture site, which can damage the buccal branch of the facial nerve during CFP. 2015 BMJ Publishing Group Ltd.

Transgenic mice expressing cyan fluorescent protein as a reporter strain to detect the effects of rotenone toxicity on retinal ganglion cells.

PubMed

Hayworth, C R; Rojas, J C; Gonzalez-Lima, F

2008-01-01

This is the first study using a reporter transgenic model to investigate the effects of an environmental toxin on the retina. Rotenone is a widely used pesticide that inhibits mitochondrial complex I and produces neurotoxicity. Previous studies demonstrated the time course and dose response of rotenone toxicity on retinal ganglion cells (RGC). However, previous analyses of rotenone-induced retinotoxicity provided little detail of the optic nerve axons and cellular pathology. These limitations were successfully surmounted by using a transgenic mouse line shown to express cyan fluorescent protein (CFP) in neurons, including RGC, under regulatory elements of the human the thy1.1 promoter (thy-CFP). Data showed that CFP expression is limited to RGC and their processes in the retina of thy-CFP mice. Eyes exposed to the pesticide rotenone displayed marked alterations in RGC morphology, inner plexiform layer, optic disc, and optic nerves. After 24 h, the number of CFP-labeled RGC was reduced 50%. Correlated with a loss of RGC bodies was an approximate 50% reduction in CFP fluorescence intensity at the optic disc. The findings showed that rotenone-induced degeneration of RGC and their processes can be visualized with exquisite detail in thy-CFP mice, and that this approach may provide a novel and effective way to monitor the association between environmental toxins and neurodegeneration in living animals.

Use of Surveillance Systems in Detection of a Ciguatera Fish Poisoning Outbreak - Orange County, Florida, 2014.

PubMed

Klekamp, Benjamin G; Bodager, Dean; Matthews, Sarah D

2015-10-16

What is already known on this topic? Ciguatera fish poisoning (CFP), caused by the ingestion of predatory reef-dwelling fish harboring ciguatoxins is one of the most commonly reported fish-associated marine intoxications. Ciguatoxin retains toxicity regardless of freezing or cooking. Prompt treatment can reduce debilitating neurologic symptoms that are associated with CFP.What is added by this report? Syndromic surveillance systems in Florida identified six adults with CFP following consumption of black grouper. Five patients sought medical attention; health care providers did not make a diagnosis of CFP or report the cases to public health authorities, and none of the patients received treatment. Close collaboration among several investigating agencies allowed traceback efforts to link black grouper consumed by all patients to a common international distributor.What are the implications for public health practice? Syndromic surveillance systems capable of detecting CFP are essential public health tools to identify outbreaks and enhance investigations. Medical and public health practitioners should be educated to inquire about recent fish consumption when evaluating patients with clinically compatible signs and symptoms to allow for prompt treatment, and report suspected CFP cases to public health authorities to facilitate source-food traceback efforts. Public education on avoidance of consumption of relatively large predatory reef fish species known to be from ciguatoxic-endemic areas might reduce the risk for CFP.

CFA or CFP: A Guide for Professors

ERIC Educational Resources Information Center

Moy, Ronald L.

2011-01-01

The CFA Institute and the CFP Board of Standards provide professional certifications in the field of finance. In this paper, I provide my experience with the CFA and CFP programs in order to give other professors some insight into the process of attaining the designations. I hope to provide answers to some of the questions that other faculty…

Estimation of Cloud Fraction Profile in Shallow Convection Using a Scanning Cloud Radar

DOE PAGES

Oue, Mariko; Kollias, Pavlos; North, Kirk W.; ...

2016-10-18

Large spatial heterogeneities in shallow convection result in uncertainties in estimations of domain-averaged cloud fraction profiles (CFP). This issue is addressed using large eddy simulations of shallow convection over land coupled with a radar simulator. Results indicate that zenith profiling observations are inadequate to provide reliable CFP estimates. Use of Scanning Cloud Radar (SCR), performing a sequence of cross-wind horizon-to-horizon scans, is not straightforward due to the strong dependence of radar sensitivity to target distance. An objective method for estimating domain-averaged CFP is proposed that uses observed statistics of SCR hydrometeor detection with height to estimate optimum sampling regions. Thismore » method shows good agreement with the model CFP. Results indicate that CFP estimates require more than 35 min of SCR scans to converge on the model domain average. Lastly, the proposed technique is expected to improve our ability to compare model output with cloud radar observations in shallow cumulus cloud conditions.« less

Review of life-cycle approaches coupled with data envelopment analysis: launching the CFP + DEA method for energy policy making.

PubMed

Vázquez-Rowe, Ian; Iribarren, Diego

2015-01-01

Life-cycle (LC) approaches play a significant role in energy policy making to determine the environmental impacts associated with the choice of energy source. Data envelopment analysis (DEA) can be combined with LC approaches to provide quantitative benchmarks that orientate the performance of energy systems towards environmental sustainability, with different implications depending on the selected LC + DEA method. The present paper examines currently available LC + DEA methods and develops a novel method combining carbon footprinting (CFP) and DEA. Thus, the CFP + DEA method is proposed, a five-step structure including data collection for multiple homogenous entities, calculation of target operating points, evaluation of current and target carbon footprints, and result interpretation. As the current context for energy policy implies an anthropocentric perspective with focus on the global warming impact of energy systems, the CFP + DEA method is foreseen to be the most consistent LC + DEA approach to provide benchmarks for energy policy making. The fact that this method relies on the definition of operating points with optimised resource intensity helps to moderate the concerns about the omission of other environmental impacts. Moreover, the CFP + DEA method benefits from CFP specifications in terms of flexibility, understanding, and reporting.

Review of Life-Cycle Approaches Coupled with Data Envelopment Analysis: Launching the CFP + DEA Method for Energy Policy Making

PubMed Central

Vázquez-Rowe, Ian

2015-01-01

Life-cycle (LC) approaches play a significant role in energy policy making to determine the environmental impacts associated with the choice of energy source. Data envelopment analysis (DEA) can be combined with LC approaches to provide quantitative benchmarks that orientate the performance of energy systems towards environmental sustainability, with different implications depending on the selected LC + DEA method. The present paper examines currently available LC + DEA methods and develops a novel method combining carbon footprinting (CFP) and DEA. Thus, the CFP + DEA method is proposed, a five-step structure including data collection for multiple homogenous entities, calculation of target operating points, evaluation of current and target carbon footprints, and result interpretation. As the current context for energy policy implies an anthropocentric perspective with focus on the global warming impact of energy systems, the CFP + DEA method is foreseen to be the most consistent LC + DEA approach to provide benchmarks for energy policy making. The fact that this method relies on the definition of operating points with optimised resource intensity helps to moderate the concerns about the omission of other environmental impacts. Moreover, the CFP + DEA method benefits from CFP specifications in terms of flexibility, understanding, and reporting. PMID:25654136

Simulating flow in karst aquifers at laboratory and sub-regional scales using MODFLOW-CFP

NASA Astrophysics Data System (ADS)

Gallegos, Josue Jacob; Hu, Bill X.; Davis, Hal

2013-12-01

Groundwater flow in a well-developed karst aquifer dominantly occurs through bedding planes, fractures, conduits, and caves created by and/or enlarged by dissolution. Conventional groundwater modeling methods assume that groundwater flow is described by Darcian principles where primary porosity (i.e. matrix porosity) and laminar flow are dominant. However, in well-developed karst aquifers, the assumption of Darcian flow can be questionable. While Darcian flow generally occurs in the matrix portion of the karst aquifer, flow through conduits can be non-laminar where the relation between specific discharge and hydraulic gradient is non-linear. MODFLOW-CFP is a relatively new modeling program that accounts for non-laminar and laminar flow in pipes, like karst caves, within an aquifer. In this study, results from MODFLOW-CFP are compared to those from MODFLOW-2000/2005, a numerical code based on Darcy's law, to evaluate the accuracy that CFP can achieve when modeling flows in karst aquifers at laboratory and sub-regional (Woodville Karst Plain, Florida, USA) scales. In comparison with laboratory experiments, simulation results by MODFLOW-CFP are more accurate than MODFLOW 2005. At the sub-regional scale, MODFLOW-CFP was more accurate than MODFLOW-2000 for simulating field measurements of peak flow at one spring and total discharges at two springs for an observed storm event.

Detection of Mitochondrial Caspase Activity in Real Time In Situ in Live Cells

NASA Astrophysics Data System (ADS)

Zhang, Yingpei; Haskins, Catherine; Lopez-Cruzan, Marisa; Zhang, Jianhua; Centonze, Victoria E.; Herman, Brian

2004-08-01

Apoptosis plays an important role in many physiological and pathological processes. The initiation and execution of the cell death program requires activation of multiple caspases in a stringently temporal order. Here we describe a method that allows real-time observation of caspase activation in situ in live cells based on fluorescent resonance energy transfer (FRET) measurement using the prism and reflector imaging spectroscopy system (PARISS). When a fusion protein consisting of CFP connected to YFP via an intervening caspase substrate that has been targeted to a specific subcellular location is excited with a light source whose wavelength matches the cyan fluorescent protein (CFP) excitation peak, the energy absorbed by the CFP fluorophore is not emitted as fluorescence. Instead, the excitation energy is absorbed by the nearby yellow fluorescent protein (YFP) fluorophore that is covalently linked to CFP through a short peptide containing the caspase substrate. Cleavage of the linker peptide by caspases results in loss of FRET due to the separation of CFP and YFP fluorophores. Using a mitochondrially targeted CFP caspase 3 substrate YFP construct (mC3Y), we demonstrate for the first time that there is caspase-3-like activity in the mitochondrial matrix of some cells at very late stage of apoptosis.

Preparation of immunochromatographic strips for rapid detection of early secreted protein ESAT-6 and culture filtrate protein CFP-10 from Mycobacterium tuberculosis

PubMed Central

Wu, Xiaoxin; Wang, Yeping; Weng, Tianhao; Hu, Chenyu; Wang, Frederick X.C.; Wu, Zhigang; Yu, Dongshan; Lu, Huoquan; Yao, Hangping

2017-01-01

Abstract The early secreted protein early secretory antigenic target 6(ESAT-6) and the culture filtrate protein 10 (CFP-10) are 2 antigens that are specific to Mycobacterium tuberculosis. These 2 antigens are good targets for tuberculosis (TB) detection. To rapidly diagnose TB across a variety of samples, we developed colloidal gold immunochromatographic strips (ICSs) based on ESAT-6 and CFP-10. The strips were evaluated using 233 samples, including sputum, plasma, and pleural effusion samples. The positive detection rates for ICSs for ESAT-6 and CFP-10 in sputum (culture-positive for M tuberculosis) were 100% and 91.2%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in plasma were 34.1% and 29.4%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in pleural effusion were 64.7% and 55.9%, respectively. Experimental analysis of culture supernatant showing that the ICS developed for ESAT-6 had a sensitivity of 100% and a specificity of 91.2%. While the ICS developed for CFP-10 had a sensitivity of 91.2% and a specificity of 88.2%. The validity of the test is limited by source of sample. The technique is sensitive and specific for samples in sputum and culture media but not for plasma or pleural effusion samples. Detection of M tuberculosis using ICSs is rapid, simple, and relatively effective; thus, ICSs are a potential screening tool for TB. PMID:29390519

Preparation of immunochromatographic strips for rapid detection of early secreted protein ESAT-6 and culture filtrate protein CFP-10 from Mycobacterium tuberculosis.

PubMed

Wu, Xiaoxin; Wang, Yeping; Weng, Tianhao; Hu, Chenyu; Wang, Frederick X C; Wu, Zhigang; Yu, Dongshan; Lu, Huoquan; Yao, Hangping

2017-12-01

The early secreted protein early secretory antigenic target 6(ESAT-6) and the culture filtrate protein 10 (CFP-10) are 2 antigens that are specific to Mycobacterium tuberculosis. These 2 antigens are good targets for tuberculosis (TB) detection.To rapidly diagnose TB across a variety of samples, we developed colloidal gold immunochromatographic strips (ICSs) based on ESAT-6 and CFP-10.The strips were evaluated using 233 samples, including sputum, plasma, and pleural effusion samples.The positive detection rates for ICSs for ESAT-6 and CFP-10 in sputum (culture-positive for M tuberculosis) were 100% and 91.2%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in plasma were 34.1% and 29.4%, respectively. The positive detection rates for ICSs for ESAT-6 and CFP-10 in pleural effusion were 64.7% and 55.9%, respectively. Experimental analysis of culture supernatant showing that the ICS developed for ESAT-6 had a sensitivity of 100% and a specificity of 91.2%. While the ICS developed for CFP-10 had a sensitivity of 91.2% and a specificity of 88.2%.The validity of the test is limited by source of sample. The technique is sensitive and specific for samples in sputum and culture media but not for plasma or pleural effusion samples. Detection of M tuberculosis using ICSs is rapid, simple, and relatively effective; thus, ICSs are a potential screening tool for TB. Copyright © 2017 The Authors. Published by Wolters Kluwer Health, Inc. All rights reserved.

Assessment of a Protein Cocktail-Based Skin Test for Bovine Tuberculosis in a Double-Blind Field Test in Cattle

PubMed Central

Xin, Ting; Jia, Hong; Ding, Jiabo; Li, Pingjun; Yang, Hongjun; Hou, Shaohua; Yuan, Weifeng; Guo, Xiaoyu; Wang, Haichun; Liang, Qianqian; Li, Ming

2013-01-01

Bovine tuberculosis (bTB) is a worldwide zoonosis caused mainly by Mycobacterium bovis. The traditional diagnostic method used often is the tuberculin skin test, which uses bovine purified protein derivatives (PPD-B). However, it is difficult to maintain uniformity of PPD-B from batch to batch, and it shares common antigens with nonpathogenic environmental mycobacteria. To overcome these problems, M. bovis-specific antigens that showed good T cell stimulation, such as CFP-10, ESAT-6, Rv3615c, etc., have been used in the skin test, but there have been no large-scale clinical studies on these antigens. In this study, two combinations (CFP-10/ESAT-6/TB10.4 protein cocktail and CFP-10/ESAT-6/Rv3872/MPT63 protein cocktail) were developed and used as stimuli in the skin test. Cattle were double-blind tested to assess the efficiency of the protein cocktail-based skin tests. The results showed that the CFP-10/ESAT-6/TB10.4 protein cocktail-based skin test can differentiate TB-infected cattle from Mycobacterium avium-infected ones and that it shows a high degree of agreement with the traditional tuberculin skin test (κ = 0.8536) and gamma interferon (IFN-γ) release assay (κ = 0.8154). Compared to the tuberculin skin test, the relative sensitivity and relative specificity of the CFP-10/ESAT-6/TB10.4-based skin test were 87% and 97%, respectively., The relative sensitivity and relative specificity of the CFP-10/ESAT-6/TB10.4-based skin test were 93% and 92%, respectively, on comparison with the IFN-γ release assay. The correlation between the increases in skin thickness observed after the inoculation of stimuli was high (PPD-B versus CFP-10/ESAT-6/TB10.4, Spearman r of 0.8435). The correlation between the optical density at 450 nm (OD450) obtained after blood stimulation with PPD-B and the increase in skin thickness observed after inoculation of the CFP-10/ESAT-6/TB10.4 protein cocktail was high (Spearman r = 0.7335). Therefore, the CFP-10/ESAT-6/TB10.4-based skin test responses correlate to traditional measures of bovine TB evaluation, including skin test and gamma interferon release assay. PMID:23365203

Fused Mycobacterium tuberculosis multi-stage immunogens with an Fc-delivery system as a promising approach for the development of a tuberculosis vaccine.

PubMed

Mosavat, Arman; Soleimanpour, Saman; Farsiani, Hadi; Sadeghian, Hamid; Ghazvini, Kiarash; Sankian, Mojtaba; Jamehdar, Saeid Amel; Rezaee, Seyed Abdolrahim

2016-04-01

Tuberculosis (TB) remains a major health problem worldwide. Currently, the Bacilli Calmette-Guérin (BCG) is the only available licensed TB vaccine, which has low efficacy in protection against adult pulmonary TB. Therefore, the development of a safe and effective vaccine against TB needs global attention. In the present study, a novel multi-stage subunit vaccine candidate from culture filtrate protein-10 (CFP-10) and heat shock protein X (HspX) of Mycobacterium tuberculosis fused to the Fc domain of mouse IgG2a as a selective delivery system for antigen-presenting cells (APCs) was produced and its immunogenicity assessed. The optimized gene constructs were introduced into pPICZαA expression vectors, and the resultant plasmids (pPICZαA-CFP-10:Hspx:Fcγ2a and pPICZαA-CFP-10:Hspx:His) were transferred into Pichia pastoris by electroporation. The identification of both purified recombinant fusion proteins was evaluated by SDS-PAGE and immunoblotting. Then the immunogenicity of the recombinant proteins with and without BCG was evaluated in BALB/c mice by assessing the level of IFN-γ, IL-12, IL-4, IL-17 and TGF-β cytokines. Both multi-stage vaccines (CFP-10:HspX:Fcγ2a and CFP-10:HspX:His) induced Th1-type cellular responses by producing high level of IFN-γ (272 pg/mL, p<0.001) and IL-12 (191 pg/mL, p<0.001). However, the Fc-tagged recombinant protein induced more effective Th1-type cellular responses with a low level of IL-4 (10 pg/mL) compared to the CFP-10:HspX:His group. The production of IFN-γ to CFP-10:HspX:Fcγ2a was markedly consistent and showed an increasing trend for IL-12 compared with the BCG or CFP-10:HspX:His primed and boosted groups. Findings revealed that CFP-10:Hspx:Fcγ2a fusion protein can elicit strong Th1 antigen-specific immune responses in favor of protective immunity in mice and could provide new insight for introducing an effective multi-stage subunit vaccine against TB. Copyright © 2016 Elsevier B.V. All rights reserved.

Kepler Certified False Positive Table

NASA Technical Reports Server (NTRS)

Bryson, Stephen T.; Batalha, Natalie Marie; Colon, Knicole Dawn; Coughlin, Jeffrey Langer; Haas, Michael R.; Henze, Chris; Huber, Daniel; Morton, Tim; Rowe, Jason Frank; Mullally, Susan Elizabeth;

Facile synthesis of Co(OH)2/Al(OH)3 nanosheets with improved electrochemical properties for asymmetric supercapacitor

NASA Astrophysics Data System (ADS)

Zhao, Cuimei; Ren, Fang; Cao, Yang; Xue, Xiangxin; Duan, Xiaoyue; Wang, Hairui; Chang, Limin

2018-01-01

Sheet-like Co(OH)2/Al(OH)3 or Co(OH)2 nanomaterial has been synthesized on conducting carbon fiber paper (CFP) by a facile one-step electrochemical deposition. The binder-free Co(OH)2/Al(OH)3/CFP displays an improved electrical conductivity, electrochemical activity and material utilization than solitary Co(OH)2, therefore Co(OH)2/Al(OH)3 nanomaterial exhibits improved electrochemical properties (a maximum capacitance of 1006 Fg-1 at 2 Ag-1, with 77% retention even at a high current density of 32 Ag-1, and more than 87% of the capacitance retention after 10000 cycles at 32 Ag-1) in comparison to that of the Co(OH)2/CFP (709 Fg-1, 65%, 79%). In addition, an asymmetric supercapacitor (ASC) fabricated with Co(OH)2/Al(OH)3/CFP positive electrode and AC/CFP negative electrode demonstrates ultrahigh specific capacitance (75.8 Fg-1) and potential window (1.7 V). These encouraging results make these low-cost and eco-friendly materials promising for high-performance energy storage application.

Catalytic fast co-pyrolysis of biomass and food waste to produce aromatics: Analytical Py-GC/MS study.

PubMed

Zhang, Bo; Zhong, Zhaoping; Min, Min; Ding, Kuan; Xie, Qinglong; Ruan, Roger

2015-01-01

In this study, catalytic fast co-pyrolysis (co-CFP) of corn stalk and food waste (FW) was carried out to produce aromatics using quantitative pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), and ZSM-5 zeolite in the hydrogen form was employed as the catalyst. Co-CFP temperature and a parameter called hydrogen to carbon effective ratio (H/C(eff) ratio) were examined for their effects on the relative content of aromatics. Experimental results showed that co-CFP temperature of 600 °C was optimal for the formation of aromatics and other organic pyrolysis products. Besides, H/C(eff) ratio had an important influence on product distribution. The yield of total organic pyrolysis products and relative content of aromatics increased non-linearly with increasing H/C(eff) ratio. There was an apparent synergistic effect between corn stalk and FW during co-CFP process, which promoted the production of aromatics significantly. Co-CFP of biomass and FW was an effective method to produce aromatics and other petrochemicals. Copyright © 2015. Published by Elsevier Ltd.

Significance of immune response to Mycobacterium tuberculosis culture filtrate protein antigens in cerebrospinal fluid of tuberculous meningitis patients: A search for diagnostic marker.

PubMed

Giribhattanavar, Prashant; Kumar, Kavitha; Raajasekar, Siddarth; Chandrashekar, Nagarathna; Patil, Shripad A

2017-01-01

Mycobacterium tuberculosis (H37Ra) culture filtrate proteins (CFP) are explored as a diagnostic marker for tuberculous meningitis (TBM). Cerebrospinal fluid (CSF) samples from patients were categorized as confirmed (n = 47), suspected (n = 20), and non-TBM (n = 25) cases. Immune response by Western blot revealed TBM CSF samples are having heterogeneous response to CFP. CFP ELISA was 92% sensitive and 38.30% specific. ODs of confirmed TBM and non-TBM cases were significantly different (P < 0.0001) and also the suspected TBM and non-TBM cases (P = 0.0001). No significant difference noticed in TBM and suspected TBM (P = 0.90). Thus, CFP can be a better biomarker for the diagnosis of TBM.

Mycobacteria-specific cytokine responses as correlates of treatment response in active and latent tuberculosis.

PubMed

Clifford, Vanessa; Tebruegge, Marc; Zufferey, Christel; Germano, Susie; Forbes, Ben; Cosentino, Lucy; McBryde, Emma; Eisen, Damon; Robins-Browne, Roy; Street, Alan; Denholm, Justin; Curtis, Nigel

2017-08-01

A biomarker indicating successful tuberculosis (TB) therapy would assist in determining appropriate length of treatment. This study aimed to determine changes in mycobacteria-specific antigen-induced cytokine biomarkers in patients receiving therapy for latent or active TB, to identify biomarkers potentially correlating with treatment success. A total of 33 adults with active TB and 36 with latent TB were followed longitudinally over therapy. Whole blood stimulation assays using mycobacteria-specific antigens (CFP-10, ESAT-6, PPD) were done on samples obtained at 0, 1, 3, 6 and 9 months. Cytokine responses (IFN-γ, IL-1ra, IL-2, IL-10, IL-13, IP-10, MIP-1β, and TNF-α) in supernatants were measured by Luminex xMAP immunoassay. In active TB cases, median IL-1ra (with CFP-10 and with PPD stimulation), IP-10 (CFP-10, ESAT-6), MIP-1β (ESAT-6, PPD), and TNF-α (ESAT-6) responses declined significantly over the course of therapy. In latent TB cases, median IL-1ra (CFP-10, ESAT-6, PPD), IL-2 (CFP-10, ESAT-6), and IP-10 (CFP-10, ESAT-6) responses declined significantly. Mycobacteria-specific cytokine responses change significantly over the course of therapy, and their kinetics in active TB differ from those observed in latent TB. In particular, mycobacteria-specific IL-1ra responses are potential correlates of successful therapy in both active and latent TB. Copyright © 2017 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

Prototype ultra wideband-based wireless body area network--consideration of CAP and CFP slot allocation during human walking motion.

PubMed

Takei, Yuichiro; Katsuta, Hiroki; Takizawa, Kenichi; Ikegami, Tetsushi; Hamaguchi, Kiyoshi

2012-01-01

This paper presents an experimental evaluation of communication during human walking motion, using the medium access control (MAC) evaluation system for a prototype ultra-wideband (UWB) based wireless body area network for suitable MAC parameter settings for data transmission. Its physical layer and MAC specifications are based on the draft standard in IEEE802.15.6. This paper studies the effects of the number of retransmissions and the number of commands of GTS (guaranteed time slot) request packets in the CAP (contention access period) during human walking motion by varying the number of sensor nodes or the number of CFP (contention free period) slots in the superframe. The experiments were performed in an anechoic chamber. The number of packets received is decreased by packet loss caused by human walking motion in the case where 2 slots are set for CFP, regardless of the number of nodes, and this materially decreases the total number of packets received. The number of retransmissions and the GTS request commands increase according to increases in the number of nodes, largely reflecting the effects of the number of CFP slots in the case where 4 nodes are attached. In the cases where 2 or 3 nodes are attached and 4 slots are set for CFP, the packet transmission rate is more than 95%. In the case where 4 nodes are attached and 6 slots are set for CFP, the packet transmission rate is reduced to 88% at best.

Visualizing Breast Cancer Cell Interaction with Tumor-Infiltrating Lymphocytes During Immunotherapy

DTIC Science & Technology

2013-04-01

in order to assess the motility cells in this tissue. To do that, CXCR6 GFP/+ mice were injected with 4T1 CFP cells, on day 30, metastasis were...Figure1: Imaging of metastasis A: Image of a metastase at day 30 after tumor implantation. 4T1-CFP tumor cells (blue), CXCR6 GFP/+ infiltrating...recognition or not. Figure 2 : Motility of CXCR6 -GFP+ cells in the periphery and core of the tumor. A-Image of 4T1-CFP tumor cells (blue), CXCR6 GFP

Ternary blends containing demercurated lighting phosphor and MSWI fly ash as high-performance binders for stabilizing and recycling electroplating sludge.

PubMed

Huang, Wu-Jang; Wu, Chia-Teng; Wu, Chang-En; Hsieh, Lin-Huey; Li, Chang-Chien; Lain, Chi-Yuan; Chu, Wei

2008-08-15

This paper describes the solidification and stabilization of electroplating sludge treated with a high-performance binder made from portland type-I cement, municipal solid waste incineration fly ash, and lighting phosphor powder (called as cement-fly ash-phosphor binder, CFP). The highest 28-day unconfined compressive strength of the CFP-treated paste was 816 kg/cm(2) at a ratio of cement to fly ash to lighting phosphor powder of 90:5:5; the strength of this composition also fulfilled the requirement of a high-strength concrete (>460 kg/cm(2) at 28 days). The CFP-stabilized sludge paste samples passed the Taiwanese EPA toxicity characteristic leaching procedure test and, therefore, could be used either as a building material or as a controlled low-strength material, depending on the sludge-to-CFP binder ratio.

Sixth Annual Conference on Computers, Freedom, and Privacy: The RealAudio Proceedings.

ERIC Educational Resources Information Center

Glover, Barbara; Meernik, Mary

1996-01-01

Reviews the sixth Conference on Computers, Freedom, and Privacy (CFP) held in March 1996. Highlights include the Communications Decency Act, part of the 1996 Telecommunications Reform Act; European views; Internet service providers; limiting online speech on campus; cryptography; the global information infrastructure; copyright; and China and the…

Cingulate, Frontal and Parietal Cortical Dysfunction in Attention-Deficit/Hyperactivity Disorder

PubMed Central

Bush, George

2011-01-01

Functional and structural neuroimaging have identified abnormalities of the brain that are likely to contribute to the neuropathophysiology of attention-deficit/hyperactivity disorder (ADHD). In particular, hypofunction of the brain regions comprising the cingulo-frontal-parietal (CFP) cognitive-attention network have been consistently observed across studies. These are major components of neural systems that are relevant to ADHD, including cognitive/attention networks, motor systems and reward/feedback-based processing systems. Moreover, these areas interact with other brain circuits that have been implicated in ADHD, such as the “default mode” resting state network. ADHD imaging data related to CFP network dysfunction will be selectively highlighted here to help facilitate its integration with the other information presented in this special issue. Together, these reviews will help shed light on the neurobiology of ADHD. PMID:21489409

Morphology controlled MnO2 electrodeposited on carbon fiber paper for high-performance supercapacitors

NASA Astrophysics Data System (ADS)

Ye, Zhiguo; Li, Tao; Ma, Guang; Peng, Xinyuan; Zhao, Jun

2017-05-01

Four different morphologies of nanostructured MnO2 (nanospheres, nanosheets, nanoflowers and nanonods) were fabricated on a carbon fiber paper (CFP) substrate using a facile method of anodic electrodeposition by varying the H2SO4 concentration and current density. The fabricated composite electrodes were characterized by field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD) and electrochemical techniques. The composite electrodes with MnO2 nanosphere/CFP, MnO2 nanosheet/CFP, MnO2 nanoflower/CFP and MnO2 nanonod/CFP achieved a relatively high specific capacitance (areal capacitance) of 134.4 F g-1 (0.20 F cm-2), 226.3 F g-1 (0.33 F cm-2), 235.6 F g-1 (0.35 F cm-2) and 362.5 F g-1 (0.54 F cm-2) at 0.5 A g-1, respectively. When the GV charging-discharging rate increased from 0.5 to 5 A g-1, the MnO2 nanorod/CFP composite decreased from 362.5 F g-1 (0.54 F cm-2) to 160.0 F g-1 (0.24 F cm-2), which is a relatively high retention of the original capacitance (i.e., 44.1%). All the composite electrodes with various nanostructured MnO2 morphologies under flat and bent states retained more than 95% and 90% of the initial capacitance after 5000 cycles at 5 A g-1, respectively, which demonstrates outstanding cycling stability. This study provides a novel approach for high-performance, morphology-controllable metal oxide electrodes for supercapacitors.

Antiobesity Effects of Sansa (Crataegi fructus) on 3T3-L1 Cells and on High-Fat-High-Cholesterol Diet-Induced Obese Rats.

PubMed

Lee, Jae-Joon; Lee, Hyun-Joo; Oh, Seon-Woo

2017-01-01

This study was performed to investigate the effects of Crataegi fructus ethanol extracts (CFEEs) on the differentiation of 3T3-L1 cells, and to evaluate the effects of C. fructus powder (CFP) on lipid metabolism and its antiobesity effect in rats fed a high-fat and high-cholesterol (HFC) diet. Both in vitro and in vivo studies were performed for physiological activity and antiobesity effects on the serum, liver, and adipose tissues in obesity-induced rats. CFEEs showed significant inhibitory action on differentiation and triglyceride (TG) accumulation in 3T3-L1 mature cells in a dose-dependent manner. Subcutaneous, mesenteric, epididymal, and total adipose tissue weights of HFC diet group were heavier than those of normal diet (N) group, whereas those of groups fed CFP were significantly decreased. Levels of serum TGs, total cholesterol (TC), and low-density lipoprotein cholesterol were significantly decreased in the CFP groups than in the HFC group, whereas the serum high-density lipoprotein cholesterol level decreased in the HFC group and markedly increased in the CFP groups. TC and TG levels in the liver and adipose tissues were significantly lower in CFP groups than in the HFC groups. In addition, feeding with CFP significantly reduced the occurrence of fatty liver deposits and steatosis, and inhibited an HFC diet-induced increase in adipocyte size. These results suggest that C. fructus may improve lipid metabolism in the serum, liver, and adipose tissue, and may potentially reduce lipid storage.

Numerical simulation of freshwater/seawater interaction in a dual-permeability karst system with conduits: the development of discrete-continuum VDFST-CFP model

NASA Astrophysics Data System (ADS)

Xu, Zexuan; Hu, Bill

2016-04-01

Dual-permeability karst aquifers of porous media and conduit networks with significant different hydrological characteristics are widely distributed in the world. Discrete-continuum numerical models, such as MODFLOW-CFP and CFPv2, have been verified as appropriate approaches to simulate groundwater flow and solute transport in numerical modeling of karst hydrogeology. On the other hand, seawater intrusion associated with fresh groundwater resources contamination has been observed and investigated in numbers of coastal aquifers, especially under conditions of sea level rise. Density-dependent numerical models including SEAWAT are able to quantitatively evaluate the seawater/freshwater interaction processes. A numerical model of variable-density flow and solute transport - conduit flow process (VDFST-CFP) is developed to provide a better description of seawater intrusion and submarine groundwater discharge in a coastal karst aquifer with conduits. The coupling discrete-continuum VDFST-CFP model applies Darcy-Weisbach equation to simulate non-laminar groundwater flow in the conduit system in which is conceptualized and discretized as pipes, while Darcy equation is still used in continuum porous media. Density-dependent groundwater flow and solute transport equations with appropriate density terms in both conduit and porous media systems are derived and numerically solved using standard finite difference method with an implicit iteration procedure. Synthetic horizontal and vertical benchmarks are created to validate the newly developed VDFST-CFP model by comparing with other numerical models such as variable density SEAWAT, couplings of constant density groundwater flow and solute transport MODFLOW/MT3DMS and discrete-continuum CFPv2/UMT3D models. VDFST-CFP model improves the simulation of density dependent seawater/freshwater mixing processes and exchanges between conduit and matrix. Continuum numerical models greatly overestimated the flow rate under turbulent flow condition but discrete-continuum models provide more accurate results. Parameters sensitivities analysis indicates that conduit diameter and friction factor, matrix hydraulic conductivity and porosity are important parameters that significantly affect variable-density flow and solute transport simulation. The pros and cons of model assumptions, conceptual simplifications and numerical techniques in VDFST-CFP are discussed. In general, the development of VDFST-CFP model is an innovation in numerical modeling methodology and could be applied to quantitatively evaluate the seawater/freshwater interaction in coastal karst aquifers. Keywords: Discrete-continuum numerical model; Variable density flow and transport; Coastal karst aquifer; Non-laminar flow

Preparation of wafer-level glass cavities by a low-cost chemical foaming process (CFP).

PubMed

Shang, Jintang; Chen, Boyin; Lin, Wei; Wong, Ching-Ping; Zhang, Di; Xu, Chao; Liu, Junwen; Huang, Qing-An

2011-04-21

A novel foaming process-chemical foaming process (CFP)-using foaming agents to fabricate wafer-level micro glass cavities including channels and bubbles was investigated. The process consists of the following steps sequentially: (1) shallow cavities were fabricated by a wet etching on a silicon wafer; (2) powders of a proper foaming agent were placed in a silicon cavity, named 'mother cavity', on the etched silicon surface; (3) the silicon cavities were sealed with a glass wafer by anodic bonding; (4) the bonded wafers were heated to above the softening point of the glass, and baked for several minutes, when the gas released by the decomposition of the foaming agent in the 'mother cavity' went into the other sealed interconnected silicon cavities to foam the softened glass into cylindrical channels named 'daughter channels', or spherical bubbles named 'son bubbles'. Results showed that wafer-level micro glass cavities with smooth wall surfaces were achieved successfully without contamination by the CFP. A model for the CFP was proposed to predict the final shape of the glass cavity. Experimental results corresponded with model predictions. The CFP provides a low-cost avenue to preparation of micro glass cavities of high quality for applications such as micro-reactors, micro total analysis systems (μTAS), analytical and bio-analytical applications, and MEMS packaging.

Two clusters of ciguatera fish poisoning in Paris, France, related to tropical fish imported from the French Caribbean by travelers.

PubMed

Epelboin, Loïc; Pérignon, Alice; Hossen, Virginie; Vincent, Renaud; Krys, Sophie; Caumes, Eric

2014-01-01

Ciguatera fish poisoning (CFP) is a food-borne illness due to the consumption of reef fish containing pathogenic toxins. CFP is endemic to tropical areas and may be described in travelers in non-endemic areas. We describe two clusters of autochthonous cases of CFP in Paris, France. They were related to two fish caught in Guadeloupe (French West Indies) and consumed in Paris after being air-transported in a cooler. In both cases, fish flesh was analyzed and the presence of ciguatoxins by mouse bioassay (MBA) was confirmed. The first cluster involved eight individuals among whom five presented gastrointestinal symptoms and four presented neurological symptoms after consuming barracuda flesh (Sphyraena barracuda). The second cluster involved a couple who consumed a grey snapper (Lutjanus griseus). Most of them consulted at different emergency departments in the region of Paris. CFP may be seen in non-traveler patients outside endemic countries resulting from imported species of fish. Thus, CFP may be undiagnosed as physicians are not aware of this tropical disease outside endemic countries. The detection of ciguatoxins by MBA in the French National Reference Laboratory is useful in the confirmation of the diagnosis. © 2014 International Society of Travel Medicine.

Ciguatera fish poisoning and sea surface temperatures in the Caribbean Sea and the West Indies.

PubMed

Tester, Patricia A; Feldman, Rebecca L; Nau, Amy W; Kibler, Steven R; Wayne Litaker, R

2010-10-01

Ciguatera fish poisoning (CFP) is a circumtropical disease caused by ingestion of a variety of reef fish that bioaccumulate algal toxins. Distribution and abundance of the organisms that produce these toxins, chiefly dinoflagellates of the genus Gambierdiscus, are reported to correlate positively with water temperature. Consequently, there is growing concern that increasing temperatures associated with climate change could increase the incidence of CFP. This concern prompted experiments on the growth rates of six Gambierdiscus species at temperatures between 18 degrees C and 33 degrees C and the examination of sea surface temperatures in the Caribbean and West Indies for areas that could sustain rapid Gambierdiscus growth rates year-round. The thermal optimum for five of six Gambierdiscus species tested was >/=29 degrees C. Long-term SST data from the southern Gulf of Mexico indicate the number of days with sea surface temperatures >/=29 degrees C has nearly doubled (44 to 86) in the last three decades. To determine how the sea surface temperatures and Gambierdiscus growth data correlate with CFP incidences in the Caribbean, a literature review and a uniform, region-wide survey (1996-2006) of CFP cases were conducted. The highest CFP incidence rates were in the eastern Caribbean where water temperatures are warmest and least variable. Published by Elsevier Ltd.

'Too-much-of-a-good-thing'? The role of advanced eco-learning and contingency factors on the relationship between corporate environmental and financial performance.

PubMed

Latan, Hengky; Chiappetta Jabbour, Charbel Jose; Lopes de Sousa Jabbour, Ana Beatriz; Renwick, Douglas William Scott; Wamba, Samuel Fosso; Shahbaz, Muhammad

2018-08-15

Inspired by the natural-resource-based view (NRBV) theory, we attempt to shed light on a controversy which has been persistent over the last decade, concerning the relationship between corporate environmental performance (CEP) and corporate financial performance (CFP). Using the 'too-much-of-a-good-thing' (TMGT) concept, which suggests that "too much can be worse than too little," we link mixed results and consider the roles of advanced eco-learning and contingency factors in influencing the CEP-CFP relationship. Based on a sample composed of ISO 14001 certified companies in Indonesia, and analyzing the data using consistent Partial Least Squares (PLSc), we found that: the CEP-CFP relationship follows an inverted U-shape; advanced eco-learning is a significant predictor of the CEP-CFP relationship, meaning that organizations able to develop higher eco-learning capability will be better able to identify the ideal boundaries of investment in environmental performance without reducing their financial performance; and that contingency factors such as environmental strategy and firm size have a significant role in influencing the CEP-CFP relationship. The study's limitations, implications for practitioners and a future research agenda are also detailed. Copyright © 2018 Elsevier Ltd. All rights reserved.

Limitations of carbon footprint as indicator of environmental sustainability.

PubMed

Laurent, Alexis; Olsen, Stig I; Hauschild, Michael Z

2012-04-03

Greenhouse gas accountings, commonly referred to with the popular term carbon footprints (CFP), are a widely used metric of climate change impacts and the main focus of many sustainability policies among companies and authorities. However, environmental sustainability concerns not just climate change but also other environmental problems, like chemical pollution or depletion of natural resources, and the focus on CFP brings the risk of problem shifting when reductions in CFP are obtained at the expense of increase in other environmental impacts. But how real is this risk? Here, we model and analyze the life cycle impacts from about 4000 different products, technologies, and services taken from several sectors, including energy generation, transportation, material production, infrastructure, and waste management. By investigating the correlations between the CFP and 13 other impact scores, we show that some environmental impacts, notably those related to emissions of toxic substances, often do not covary with climate change impacts. In such situations, carbon footprint is a poor representative of the environmental burden of products, and environmental management focused exclusively on CFP runs the risk of inadvertently shifting the problem to other environmental impacts when products are optimized to become more "green". These findings call for the use of more broadly encompassing tools to assess and manage environmental sustainability.

Time Lapse to Colorectal Cancer: Telomere Dynamics Define the Malignant Potential of Polyps.

PubMed

Druliner, Brooke R; Ruan, Xiaoyang; Johnson, Ruth; Grill, Diane; O'Brien, Daniel; Lai, Tsung-Po; Rashtak, Shahrooz; Felmlee-Devine, Donna; Washechek-Aletto, Jill; Malykh, Andrei; Smyrk, Thomas; Oberg, Ann; Liu, Hongfang; Shay, Jerry W; Ahlquist, David A; Boardman, Lisa A

2016-09-01

Whereas few adenomas become cancer, most colorectal cancers arise from adenomas. Telomere length is a recognized biomarker in multiple cancers, and telomere maintenance mechanisms (TMM) are exploited by malignant cells. We sought to determine whether telomere length and TMM distinguish cancer-associated adenomas from those that are cancer-free. Tissues were identified as cancer-adjacent polyp (CAP)-residual adenoma contiguous with cancer-and cancer-free polyp (CFP)-adenomas without malignancy. Telomere length, TMM, and expression were measured in 102 tissues including peripheral blood leukocytes (PBLs), normal colon epithelium, adenoma, and cancer (in CAP cases) from 31 patients. Telomere length was measured in a separate cohort of 342 PBL from CAP and CFP patients. The mean differences in telomere length between normal and adenoma were greater in CAP than in CFP cases, P=0.001; telomere length in PBL was 91.7 bp greater in CAP than in CFP, P=0.007. Each 100 bp telomere increase was associated with a 1.14 (1.04-1.26) increased odds of being a CAP, P=0.0063. The polyp tissue from CAP patients had shorter telomeres and higher Telomerase reverse transcriptase (hTERT) expression compared with polyps from CFP patients, P=0.05. There was a greater degree of alternative lengthening of telomere (ALT) level difference in CFP polyps than in CAP polyps. The polyp telomere lengths of aggressive CAPs were significantly different from the polyps of non-aggressive CAPs, P=0.01. Adenomas that progress to cancer exhibit distinct telomere length and TMM profiles. We report for the first time that PBL telomeres differ in patients with polyps that become malignant, and therefore may have clinical value in adenoma risk assessment and management.

Electromagnetic interference shielding and microwave absorption properties of cobalt ferrite CoFe2O4/polyaniline composite

NASA Astrophysics Data System (ADS)

Ismail, Mukhils M.; Rafeeq, Sewench N.; Sulaiman, Jameel M. A.; Mandal, Avinandan

2018-05-01

Improvement of microwave-absorbing materials (MAMs) is the most important research area in various applications, such as in communication, radiation medical exposure, electronic warfare, air defense, and different civilian applications. Conducting polymer, polyaniline doped with para toluene sulphonic acid (PANI-PTSA) as well as cobalt ferrite (CoFe2O4) is synthesized by sol-gel method and intensely blends in different ratios. The characterization of the composite materials, CoFe2O4/PANI-PTSA (CFP1, CFP2, CFP3 and CFP4), was performed by X-ray diffraction (XRD), atomic force microscopy (AFM) and vibrating sample magnetometry (VSM). The microwave-absorbing properties' reflection loss (dB) and important parameters, such as complex relative permittivity ( ɛ r '- jɛ r ″) and complex relative permeability ( µ r '- jµ r ″) were measured in different microwave frequencies in the X-band (8.2-12.4 GHz) region. The composite material CFP3 showed a wider absorption frequency range and maximum reflection loss of - 28.4 dB (99.8% power absorption) at 8.1 GHz and - 9.6 dB (> 90% power absorption) at 11.2 GHz, and so the composite can be used as a microwave absorber; however, it can be more suitable for application in daily life for making cell phones above 9 GHz. Also the results showed that the thicker composites like CFP3 (4 mm) exhibit obviously better EMI SE as compared with the thinner ones (0.19, 0.19, 0.3 mm); this may be related to the low transmission of the EM wave from the composites.

Acute Isolated Central Facial Palsy as Manifestation of Middle Cerebral Artery Ischemia.

PubMed

Sands, Kara A; Shahripour, Reza Bavarsad; Kumar, Gyanendra; Barlinn, Kristian; Lyerly, Michael J; Haršány, Michal; Cure, Joel; Yakov, Yuri L; Alexandrov, Anne W; Alexandrov, Andrei V

2016-09-01

Isolated central facial palsy (I-CFP) is attributed to a lacunar syndrome affecting the corona radiata region or pons. We examined our acute stroke registry for patients presenting with I-CFP and localized their symptoms to a vascular lesion. Our database of consecutive patients with symptoms of acute cerebral ischemia admitted from January 2008 to December 2012 was reviewed for NIH Stroke Scale (NIHSS) scores and subcomponents. All patients with I-CFP ± dysarthria (total NIHSS ≤ 3) had contrast-enhanced MR-angiography and transcranial Doppler as standard of care. All ischemic lesions were localized by MRI within 72 hours from symptom onset. Of 2,202 patients with acute cerebral ischemia, 879 patients (35%) had NIHSS score ≤ 3 points (mean age 63 + 15 years, 46 % women). Nine patients (.4%) presented with I-CFP ± dysarthria. Of these, only 1 had a lesion in the corona radiata and patent MCA, 1 had a pontine lesion without proximal vessel occlusion (2/9, or 22%). Remaining 7 patients (78%) had flow-limiting thromboembolic mid-to-distal M1/proximal M2 MCA disease. Of these, 6 (86%) patients had a prominent early anterior temporal artery on MRA and nonlacunar ischemic lesions on MRI. Contrary to current teaching of lesion localization for an I-CFP, our study revealed the majority of acute patients presenting with this symptom had evidence of flow-limiting thromboembolic MCA disease rather than a lacunar lesion. Our findings underscore the essential role of comprehensive vascular imaging in patients presenting with I-CFP, which is commonly associated with acute flow-limiting thromboembolic MCA disease. Copyright © 2016 by the American Society of Neuroimaging.

The introduction of mentorship to Project 2000 in Wales.

PubMed

Neary, M; Phillips, R; Davies, B

1996-03-13

This study focused upon the introduction of mentors in the Common Foundation Programme (CFP) of Project 2000 (UKCC 1986) in Wales. It was commissioned by the Department of Health Research and Development Division on behalf of the Welsh Office Nursing Division. The study was policy oriented and its purpose was to inform future policy decision making through an analysis of the implementation of current policies for pre-registration education. The full title of our research project, 'The practitioner teacher: a study in the introduction of mentors in the pre-registration nurse education programme', implied that a clinically-based nurse practitioner with a designated teaching remit, fulfills a particular role (that of mentor) in the pre-registration nurse education programme. It was the nature, scope and impact of this mentor role during the initial implementation period of the CFP of Project 2000 in Wales to which this study addressed itself.

Two potential calmodulin-binding sequences in the ryanodine receptor contribute to a mobile, intra-subunit calmodulin-binding domain

PubMed Central

Huang, Xiaojun; Liu, Ying; Wang, Ruiwu; Zhong, Xiaowei; Liu, Yingjie; Koop, Andrea; Chen, S. R. Wayne; Wagenknecht, Terence; Liu, Zheng

2013-01-01

Summary Calmodulin (CaM), a 16 kDa ubiquitous calcium-sensing protein, is known to bind tightly to the calcium release channel/ryanodine receptor (RyR), and modulate RyR function. CaM binding studies using RyR fragments or synthetic peptides have revealed the presence of multiple, potential CaM-binding regions in the primary sequence of RyR. In the present study, we inserted GFP into two of these proposed CaM-binding sequences and mapped them onto the three-dimensional structure of intact cardiac RyR2 by cryo-electron microscopy. Interestingly, we found that the two potential CaM-binding regions encompassing, Arg3595 and Lys4269, respectively, are in close proximity and are adjacent to the previously mapped CaM-binding sites. To monitor the conformational dynamics of these CaM-binding regions, we generated a fluorescence resonance energy transfer (FRET) pair, a dual CFP- and YFP-labeled RyR2 (RyR2R3595-CFP/K4269-YFP) with CFP inserted after Arg3595 and YFP inserted after Lys4269. We transfected HEK293 cells with the RyR2R3595-CFP/K4269-YFP cDNA, and examined their FRET signal in live cells. We detected significant FRET signals in transfected cells that are sensitive to the channel activator caffeine, suggesting that caffeine is able to induce conformational changes in these CaM-binding regions. Importantly, no significant FRET signals were detected in cells co-transfected with cDNAs encoding the single CFP (RyR2R3595-CFP) and single YFP (RyR2K4269-YFP) insertions, indicating that the FRET signal stemmed from the interaction between R3595–CFP and K4269–YFP that are in the same RyR subunit. These observations suggest that multiple regions in the RyR2 sequence may contribute to an intra-subunit CaM-binding pocket that undergoes conformational changes during channel gating. PMID:23868982

NASA-OAI Collaborative Aerospace Research and Fellowship Program

NASA Technical Reports Server (NTRS)

Heyward, Ann O.; Kankam, Mark D.

2003-01-01

During the summer of 2003, a IO-week activity for university faculty entitled the NASA-OAI Collaborative Aerospace Research and Fellowship Program (CFP) was conducted at the NASA Glenn Research Center in collaboration with the Ohio Aerospace Institute (OAI). The objectives of CFP are: (1) to further the professional knowledge of qualified engineering and science faculty, (2) to stimulate an exchange of ideas between teaching participants and employees of NASA, (3) to enrich and refresh the research and teaching activities of participants' institutions, and (4) to contribute to the research objectives of Glenn. This report is intended primarily to summarize the research activities comprising the 2003 CFP Program at Glenn.

Quantitative analysis of recombination between YFP and CFP genes of FRET biosensors introduced by lentiviral or retroviral gene transfer

PubMed Central

Komatsubara, Akira T.; Matsuda, Michiyuki; Aoki, Kazuhiro

2015-01-01

Biosensors based on the principle of Förster (or fluorescence) resonance energy transfer (FRET) have been developed to visualize spatio-temporal dynamics of signalling molecules in living cells. Many of them adopt a backbone of intramolecular FRET biosensor with a cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) as donor and acceptor, respectively. However, there remains the difficulty of establishing cells stably expressing FRET biosensors with a YFP and CFP pair by lentiviral or retroviral gene transfer, due to the high incidence of recombination between YFP and CFP genes. To address this, we examined the effects of codon-diversification of YFP on the recombination of FRET biosensors introduced by lentivirus or retrovirus. The YFP gene that was fully codon-optimized to E.coli evaded the recombination in lentiviral or retroviral gene transfer, but the partially codon-diversified YFP did not. Further, the length of spacer between YFP and CFP genes clearly affected recombination efficiency, suggesting that the intramolecular template switching occurred in the reverse-transcription process. The simple mathematical model reproduced the experimental data sufficiently, yielding a recombination rate of 0.002–0.005 per base. Together, these results show that the codon-diversified YFP is a useful tool for expressing FRET biosensors by lentiviral or retroviral gene transfer. PMID:26290434

A bacterial reporter system for the evaluation of antisense oligodeoxynucleotides directed against human papillomavirus type 16 (HPV-16).

PubMed

Guapillo, Mario R; Márquez, Miguel A; Benítez-Hess, María L; Alvarez-Salas, Luis M

2006-07-01

Antisense oligodeoxynucleotides (AS-ODNs) are a promising alternative for the cure of many diseases because of their in vivo specificity and stability. However, AS-ODNs have a strong dependence on the target mRNA structure making necessary extensive in vivo testing. There is, therefore, a need to develop assays to rapidly evaluate in vivo ODN performance. We report a simple and inexpensive bacterial reporter system for the rapid in vivo evaluation of AS-ODNs directed against human papillomavirus type 16 (HPV-16) based on the destruction of a chimeric CFP mRNA using the reported HPV-16 nt 410-445 target. In vitro RNaseH assays confirmed target RNA accessibility after AS-ODN treatment. Expression of CFP in Escherichia coli BL21(DE3) with pGST-TSd2-CFP plasmid containing HPV-16 nt 410-445 target linked to CFP was blocked by transformed antisense PS-ODNs but not by two different scrambled ODN controls. A correlation was observed between bacterial CFP downregulation with the HPV-16 E6/E7 mRNA downregulation and the inhibition of anchorage-independent growth of HPV-16 containing cells suggesting that inhibition of HPV-16 E6/E7 expression by AS-ODNs directed against 410-445 target in cervical tumor cells can be tested in bacterial models.

Signal Regulatory Protein alpha (SIRPalpha)+ Cells in the Adaptive Response to ESAT-6/CFP-10 Protein of Tuberculous Mycobacteria

USDA-ARS?s Scientific Manuscript database

Early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10(CFP-10) are co-secreted proteins of Mycobacterium tuberculosis complex mycobacteria (includes M. bovis, the zoonotic agent of bovine tuberculosis) involved in phagolysosome escape of the bacillus and, potentially, in the eff...

Quantitative 13C NMR characterization of fast pyrolysis oils

DOE PAGES

Happs, Renee M.; Lisa, Kristina; Ferrell, III, Jack R.

2016-10-20

Quantitative 13C NMR analysis of model catalytic fast pyrolysis (CFP) oils following literature procedures showed poor agreement for aromatic hydrocarbons between NMR measured concentrations and actual composition. Furthermore, modifying integration regions based on DEPT analysis for aromatic carbons resulted in better agreement. Solvent effects were also investigated for hydrotreated CFP oil.

Quantitative 13C NMR characterization of fast pyrolysis oils

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

Happs, Renee M.; Lisa, Kristina; Ferrell, III, Jack R.

Quantitative 13C NMR analysis of model catalytic fast pyrolysis (CFP) oils following literature procedures showed poor agreement for aromatic hydrocarbons between NMR measured concentrations and actual composition. Furthermore, modifying integration regions based on DEPT analysis for aromatic carbons resulted in better agreement. Solvent effects were also investigated for hydrotreated CFP oil.

A Three-Hybrid System to Probe In Vivo Protein-Protein Interactions: Application to the Essential Proteins of the RD1 Complex of M. tuberculosis

PubMed Central

Bhalla, Kuhulika; Ghosh, Anamika; Kumar, Krishan; Kumar, Sushil; Ranganathan, Anand

2011-01-01

Background Protein-protein interactions play a crucial role in enabling a pathogen to survive within a host. In many cases the interactions involve a complex of proteins rather than just two given proteins. This is especially true for pathogens like M. tuberculosis that are able to successfully survive the inhospitable environment of the macrophage. Studying such interactions in detail may help in developing small molecules that either disrupt or augment the interactions. Here, we describe the development of an E. coli based bacterial three-hybrid system that can be used effectively to study ternary protein complexes. Methodology/Principal Findings The protein-protein interactions involved in M. tuberculosis pathogenesis have been used as a model for the validation of the three-hybrid system. Using the M. tuberculosis RD1 encoded proteins CFP10, ESAT6 and Rv3871 for our proof-of-concept studies, we show that the interaction between the proteins CFP10 and Rv3871 is strengthened and stabilized in the presence of ESAT6, the known heterodimeric partner of CFP10. Isolating peptide candidates that can disrupt crucial protein-protein interactions is another application that the system offers. We demonstrate this by using CFP10 protein as a disruptor of a previously established interaction between ESAT6 and a small peptide HCL1; at the same time we also show that CFP10 is not able to disrupt the strong interaction between ESAT6 and another peptide SL3. Conclusions/Significance The validation of the three-hybrid system paves the way for finding new peptides that are stronger binders of ESAT6 compared even to its natural partner CFP10. Additionally, we believe that the system offers an opportunity to study tri-protein complexes and also perform a screening of protein/peptide binders to known interacting proteins so as to elucidate novel tri-protein complexes. PMID:22087330

Some Secrets of Fluorescent Proteins: Distinct Bleaching in Various Mounting Fluids and Photoactivation of Cyan Fluorescent Proteins at YFP-Excitation

PubMed Central

Malkani, Naila; Schmid, Johannes A.

2011-01-01

Background The use of spectrally distinct variants of green fluorescent protein (GFP) such as cyan or yellow mutants (CFP and YFP, respectively) is very common in all different fields of life sciences, e.g. for marking specific proteins or cells or to determine protein interactions. In the latter case, the quantum physical phenomenon of fluorescence resonance energy transfer (FRET) is exploited by specific microscopy techniques to visualize proximity of proteins. Methodology/Principal Findings When we applied a commonly used FRET microscopy technique - the increase in donor (CFP)-fluorescence after bleaching of acceptor fluorophores (YFP), we obtained good signals in live cells, but very weak signals for the same samples after fixation and mounting in commercial microscopy mounting fluids. This observation could be traced back to much faster bleaching of CFP in these mounting media. Strikingly, the opposite effect of the mounting fluid was observed for YFP and also for other proteins such as Cerulean, TFP or Venus. The changes in photostability of CFP and YFP were not caused by the fixation but directly dependent on the mounting fluid. Furthermore we made the interesting observation that the CFP-fluorescence intensity increases by about 10 - 15% after illumination at the YFP-excitation wavelength – a phenomenon, which was also observed for Cerulean. This photoactivation of cyan fluorescent proteins at the YFP-excitation can cause false-positive signals in the FRET-microscopy technique that is based on bleaching of a yellow FRET acceptor. Conclusions/Significance Our results show that photostability of fluorescent proteins differs significantly for various media and that CFP bleaches significantly faster in commercial mounting fluids, while the opposite is observed for YFP and some other proteins. Moreover, we show that the FRET microscopy technique that is based on bleaching of the YFP is prone to artifacts due to photoactivation of cyan fluorescent proteins under these conditions. PMID:21490932

Some secrets of fluorescent proteins: distinct bleaching in various mounting fluids and photoactivation of cyan fluorescent proteins at YFP-excitation.

PubMed

Malkani, Naila; Schmid, Johannes A

2011-04-07

The use of spectrally distinct variants of green fluorescent protein (GFP) such as cyan or yellow mutants (CFP and YFP, respectively) is very common in all different fields of life sciences, e.g. for marking specific proteins or cells or to determine protein interactions. In the latter case, the quantum physical phenomenon of fluorescence resonance energy transfer (FRET) is exploited by specific microscopy techniques to visualize proximity of proteins. When we applied a commonly used FRET microscopy technique--the increase in donor (CFP)-fluorescence after bleaching of acceptor fluorophores (YFP), we obtained good signals in live cells, but very weak signals for the same samples after fixation and mounting in commercial microscopy mounting fluids. This observation could be traced back to much faster bleaching of CFP in these mounting media. Strikingly, the opposite effect of the mounting fluid was observed for YFP and also for other proteins such as Cerulean, TFP or Venus. The changes in photostability of CFP and YFP were not caused by the fixation but directly dependent on the mounting fluid. Furthermore we made the interesting observation that the CFP-fluorescence intensity increases by about 10-15% after illumination at the YFP-excitation wavelength--a phenomenon, which was also observed for Cerulean. This photoactivation of cyan fluorescent proteins at the YFP-excitation can cause false-positive signals in the FRET-microscopy technique that is based on bleaching of a yellow FRET acceptor. Our results show that photostability of fluorescent proteins differs significantly for various media and that CFP bleaches significantly faster in commercial mounting fluids, while the opposite is observed for YFP and some other proteins. Moreover, we show that the FRET microscopy technique that is based on bleaching of the YFP is prone to artifacts due to photoactivation of cyan fluorescent proteins under these conditions.

A Phylogenetic Re-Analysis of Groupers with Applications for Ciguatera Fish Poisoning

PubMed Central

Schoelinck, Charlotte; Hinsinger, Damien D.; Dettaï, Agnès; Cruaud, Corinne; Justine, Jean-Lou

2014-01-01

Background Ciguatera fish poisoning (CFP) is a significant public health problem due to dinoflagellates. It is responsible for one of the highest reported incidence of seafood-borne illness and Groupers are commonly reported as a source of CFP due to their position in the food chain. With the role of recent climate change on harmful algal blooms, CFP cases might become more frequent and more geographically widespread. Since there is no appropriate treatment for CFP, the most efficient solution is to regulate fish consumption. Such a strategy can only work if the fish sold are correctly identified, and it has been repeatedly shown that misidentifications and species substitutions occur in fish markets. Methods We provide here both a DNA-barcoding reference for groupers, and a new phylogenetic reconstruction based on five genes and a comprehensive taxonomical sampling. We analyse the correlation between geographic range of species and their susceptibility to ciguatera accumulation, and the co-occurrence of ciguatoxins in closely related species, using both character mapping and statistical methods. Results Misidentifications were encountered in public databases, precluding accurate species identifications. Epinephelinae now includes only twelve genera (vs. 15 previously). Comparisons with the ciguatera incidences show that in some genera most species are ciguateric, but statistical tests display only a moderate correlation with the phylogeny. Atlantic species were rarely contaminated, with ciguatera occurrences being restricted to the South Pacific. Conclusions The recent changes in classification based on the reanalyses of the relationships within Epinephelidae have an impact on the interpretation of the ciguatera distribution in the genera. In this context and to improve the monitoring of fish trade and safety, we need to obtain extensive data on contamination at the species level. Accurate species identifications through DNA barcoding are thus an essential tool in controlling CFP since meal remnants in CFP cases can be easily identified with molecular tools. PMID:25093850

A phylogenetic re-analysis of groupers with applications for ciguatera fish poisoning.

PubMed

Schoelinck, Charlotte; Hinsinger, Damien D; Dettaï, Agnès; Cruaud, Corinne; Justine, Jean-Lou

2014-01-01

Ciguatera fish poisoning (CFP) is a significant public health problem due to dinoflagellates. It is responsible for one of the highest reported incidence of seafood-borne illness and Groupers are commonly reported as a source of CFP due to their position in the food chain. With the role of recent climate change on harmful algal blooms, CFP cases might become more frequent and more geographically widespread. Since there is no appropriate treatment for CFP, the most efficient solution is to regulate fish consumption. Such a strategy can only work if the fish sold are correctly identified, and it has been repeatedly shown that misidentifications and species substitutions occur in fish markets. We provide here both a DNA-barcoding reference for groupers, and a new phylogenetic reconstruction based on five genes and a comprehensive taxonomical sampling. We analyse the correlation between geographic range of species and their susceptibility to ciguatera accumulation, and the co-occurrence of ciguatoxins in closely related species, using both character mapping and statistical methods. Misidentifications were encountered in public databases, precluding accurate species identifications. Epinephelinae now includes only twelve genera (vs. 15 previously). Comparisons with the ciguatera incidences show that in some genera most species are ciguateric, but statistical tests display only a moderate correlation with the phylogeny. Atlantic species were rarely contaminated, with ciguatera occurrences being restricted to the South Pacific. The recent changes in classification based on the reanalyses of the relationships within Epinephelidae have an impact on the interpretation of the ciguatera distribution in the genera. In this context and to improve the monitoring of fish trade and safety, we need to obtain extensive data on contamination at the species level. Accurate species identifications through DNA barcoding are thus an essential tool in controlling CFP since meal remnants in CFP cases can be easily identified with molecular tools.

Protection against bovine tuberculosis induced by oral vaccination of cattle with Mycobacterium bovis BCG is not enhanced by co-administration of mycobacterial protein vaccines.

PubMed

Wedlock, D Neil; Aldwell, Frank E; Vordermeier, H Martin; Hewinson, R Glyn; Buddle, Bryce M

2011-12-15

Mycobacterium bovis bacille Calmette-Guérin (BCG) delivered to calves by the oral route in a formulated lipid matrix has been previously shown to induce protection against bovine tuberculosis. A study was conducted in cattle to determine if a combination of a low dose of oral BCG and a protein vaccine could induce protective immunity to tuberculosis while not sensitising animals to tuberculin. Groups of calves (10 per group) were vaccinated by administering 2 × 10(7)colony forming units (CFU) of BCG orally or a combination of 2 × 10(7)CFU oral BCG and a protein vaccine comprised of M. bovis culture filtrate proteins (CFP) formulated with the adjuvants Chitin and Gel 01 and delivered by the intranasal route, or CFP formulated with Emulsigen and the TLR2 agonist Pam(3)CSK(4) and administered by the subcutaneous (s.c.) route. Two further groups were vaccinated with the CFP/Chitin/Gel 01 or CFP/Emulsigen/Pam(3)CSK(4) vaccines alone. Positive control groups were given 10(8)CFU oral BCG or 10(6)CFU s.c. BCG while a negative control group was non-vaccinated. All animals were challenged with M. bovis 15 weeks after vaccination and euthanized and necropsied at 16 weeks following challenge. Groups of cattle vaccinated with s.c. BCG, 10(8)CFU or 2 × 10(7)CFU oral BCG showed significant reductions in seven, three and four pathological or microbiological disease parameters, respectively, compared to the results for the non-vaccinated group. There was no evidence of protection in calves vaccinated with the combination of oral BCG and CFP/Emulsigen/Pam(3)CSK(4) or oral BCG and CFP/Chitin/Gel 01 or vaccinated with the protein vaccines alone. Positive responses in the comparative cervical skin test at 12 weeks after vaccination were only observed in animals vaccinated with s.c. BCG, 10(8)CFU oral BCG or a combination of 2 × 10(7)CFU oral BCG and CFP/Chitin/Gel 01. In conclusion, co-administration of a protein vaccine, administered by either systemic or mucosal routes with oral BCG did not enhance the protection conferred by administration of oral BCG alone. Copyright © 2011 Elsevier B.V. All rights reserved.

Long-term follow-up of 1217 consecutive short-stem total hip arthroplasty (THA): a retrospective single-center experience.

PubMed

Wacha, H; Domsel, G; Herrmann, E

2018-06-01

An arthroplasty registry in Germany has been recently established but long-term results for most short-stem innovations are missing. Short-stem hip arthroplasty is usually indicated in young active patients. Our indication was extended to older age groups, femoral neck fractures (FNF), and dysplasia. We evaluated all total hip arthroplasties (THAs) in this population with a collum femoris preserving stem (CFP) performed from 2003 to 2013. A consecutive cohort of 1217 CFP THAs with a mean age of 68.7 years was followed retrospectively for a median of 4.8 years (patient follow-up interquartile range from 3.0 to 6.9 years). A questionnaire, which we used in two previous studies, was answered by 89.15% of patients and included information regarding complaints, grade of satisfaction, re-operations, and dislocation. Of the 1217 patients, 77 had died. Survival of the stem and the cup was assessed using a competing risks approach according to an Aalen-Johanson estimator with revision for septic or aseptic loosening or death as a competing endpoint. Of the patients who answered the questionnaire, 92.5% had no complaints related to the procedures. In all 1217 patients, there were 43 revisions (4.2%) as follows: stem and cup revisions due to aseptic loosening of the stem (n = 10), infections (n = 6), pain (n = 4), or trauma (n = 3); cup revisions due to aseptic loosening (n = 3), dislocation (n = 5), and offset revisions (n = 12). Survivorship was 96% for the stem and 99% for the cup 9 years postoperatively. Statistical analysis confirmed a higher risk for revision in patients with a younger age (p = 0.033), male sex (p = 0.040), dysplasia (p = 0.032), and undersized or extra-large stems for stem revisions (p = 0.001) and female sex (p = 0.036) for cup revisions. FNF (p > 0.20) and age ≥ 80 years (p = 0.114) had no higher risk for loosening of the stem. Our data is also compared with the current literature, especially with the available CFP studies. The survival rate of the CFP stem was as high as 96% after 9 years of followup which compares well-to-previously published long-term survival rates. There is no higher risk for revision in patients 80 years old or older and in cases with femoral neck fractures. The CFP preserves also allowed using standard stems in the rare cases of revision.

Ciguatera Fish Poisoning in Hawai‘i and the Pacific

PubMed Central

Palmer, Wyatt R; Bienfang, Paul K

2014-01-01

Ciguatera fish poisoning (CFP) is a foodborne illness caused by fish containing ciguatoxin (CTX). The toxin is produced by the microalgae Gambierdiscus spp. which are then eaten by reef fish; humans contract the illness when eating either fish that have eaten the algae, or carnivorous fish that have eaten those fish. CTX is an odorless, tasteless, and colorless neurotoxin that blocks voltage-sensitive Na+ channels and accumulates in many tissues of the fish, especially the viscera. The illness is typically mild to moderate in severity with gastrointestinal (diarrhea, cramping, nausea, vomiting) and neurological (paraesthesias, cold allodynia, fatigue, pruritis) manifestations. Rarely, the disease can be more severe with significant neuropathic or cardiac effects such as bradycardia and hypotension. Endemic to Hawai‘i and islands throughout the Caribbean and Pacific, CFP incidence rates range from several to thousands of cases per 100,000 per year. Since fishing is important for local food supply, exportation, and recreation throughout the Pacific, CFP is medically and economically significant in these areas. We present a case of CFP from Hawai‘i to illustrate the disease, demonstrating that the diagnosis is primarily clinical, with confirmatory tests from fish samples available in some cases. Treatment is supportive and symptomatic with no disease specific remedy. The prognosis for most cases is good with a short duration of self-limited symptoms, but for some cases neurological sequelae can become chronic. With no effective treatment, education on which species of reef fish and which body parts to avoid eating is essential in the prevention of CFP. PMID:25478299

Ciguatera fish poisoning in Hawai'i and the Pacific.

PubMed

Copeland, Nathanial K; Palmer, Wyatt R; Bienfang, Paul K

2014-11-01

Ciguatera fish poisoning (CFP) is a foodborne illness caused by fish containing ciguatoxin (CTX). The toxin is produced by the microalgae Gambierdiscus spp. which are then eaten by reef fish; humans contract the illness when eating either fish that have eaten the algae, or carnivorous fish that have eaten those fish. CTX is an odorless, tasteless, and colorless neurotoxin that blocks voltage-sensitive Na(+) channels and accumulates in many tissues of the fish, especially the viscera. The illness is typically mild to moderate in severity with gastrointestinal (diarrhea, cramping, nausea, vomiting) and neurological (paraesthesias, cold allodynia, fatigue, pruritis) manifestations. Rarely, the disease can be more severe with significant neuropathic or cardiac effects such as bradycardia and hypotension. Endemic to Hawai'i and islands throughout the Caribbean and Pacific, CFP incidence rates range from several to thousands of cases per 100,000 per year. Since fishing is important for local food supply, exportation, and recreation throughout the Pacific, CFP is medically and economically significant in these areas. We present a case of CFP from Hawai'i to illustrate the disease, demonstrating that the diagnosis is primarily clinical, with confirmatory tests from fish samples available in some cases. Treatment is supportive and symptomatic with no disease specific remedy. The prognosis for most cases is good with a short duration of self-limited symptoms, but for some cases neurological sequelae can become chronic. With no effective treatment, education on which species of reef fish and which body parts to avoid eating is essential in the prevention of CFP.

VASCULAR ABNORMALITIES IN DIABETIC RETINOPATHY ASSESSED WITH SWEPT-SOURCE OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY WIDEFIELD IMAGING.

PubMed

Schaal, Karen B; Munk, Marion R; Wyssmueller, Iris; Berger, Lieselotte E; Zinkernagel, Martin S; Wolf, Sebastian

2017-11-10

To detect vascular abnormalities in diabetic retinopathy using swept-source optical coherence tomography angiography (SS-OCTA) widefield images, and to compare the findings with color fundus photographs (CFPs) using Early Treatment Diabetic Retinopathy Study severity grading. 3 mm × 3 mm and 12 mm × 12 mm scans were acquired to cover 70° to 80° of the posterior pole using a 100-kHz SS-OCTA instrument. Two masked graders assessed the presence of vascular abnormalities on SS-OCTA and the Early Treatment Diabetic Retinopathy Study level on CFP. The grading results were then compared. A total of 120 diabetic eyes (60 patients) were imaged with the SS-OCTA instrument. Cohort 1 (91 eyes; SS-OCTA grading only) showed microaneurysms in 91% (n = 83), intraretinal microvascular abnormalities in 79% (n = 72), and neovascularization in 21% (n = 19) of cases. Cohort 2 (52 eyes; CFP grading compared with SS-OCTA) showed microaneurysms on CFP in 90% (n = 47) and on SS-OCTA in 96% (n = 50) of cases. Agreement in intraretinal microvascular abnormality detection was fair (k = 0.2). Swept-source optical coherence tomography angiography detected 50% of intraretinal microvascular abnormality cases (n = 26), which were missed on CFP. Agreement in detecting neovascularization was moderate (k = 0.5). Agreement in detection of diabetic retinopathy features on CFP and SS-OCTA varies depending on the vascular changes examined. Swept-source optical coherence tomography angiography shows a higher detection rate of intraretinal microvascular abnormalities (P = 0.039), compared with Early Treatment Diabetic Retinopathy Study grading.

Alkaline resistant phosphate glasses and method of preparation and use thereof

DOEpatents

Brow, Richard K.; Reis, Signo T.; Velez, Mariano; Day, Delbert E.

2010-01-26

A substantially alkaline resistant calcium-iron-phosphate (CFP) glass and methods of making and using thereof. In one application, the CFP glass is drawn into a fiber and dispersed in cement to produce glass fiber reinforced concrete (GFRC) articles having the high compressive strength of concrete with the high impact, flexural and tensile strength associated with glass fibers.

Water Flow in Karst Aquifer Considering Dynamically Variable Saturation Conduit

NASA Astrophysics Data System (ADS)

Tan, Chaoqun; Hu, Bill X.

2017-04-01

The karst system is generally conceptualized as dual-porosity system, which is characterized by low conductivity and high storage continuum matrix and high conductivity and quick flow conduit networks. And so far, a common numerical model for simulating flow in karst aquifer is MODFLOW2005-CFP, which is released by USGS in 2008. However, the steady-state approach for conduit flow in CFP is physically impractical when simulating very dynamic hydraulics with variable saturation conduit. So, we adopt the method proposed by Reimann et al. (2011) to improve current model, in which Saint-Venant equations are used to model the flow in conduit. Considering the actual background that the conduit is very big and varies along flow path and the Dirichlet boundary varies with rainfall in our study area in Southwest China, we further investigate the influence of conduit diameter and outflow boundary on numerical model. And we also analyze the hydraulic process in multi-precipitation events. We find that the numerical model here corresponds well with CFP for saturated conduit, and it could depict the interaction between matrix and conduit during very dynamic hydraulics pretty well compare with CFP.

Development of a high spectral resolution lidar based on confocal Fabry-Perot spectral filters.

PubMed

Hoffman, David S; Repasky, Kevin S; Reagan, John A; Carlsten, John L

2012-09-01

The high spectral resolution lidar (HSRL) instrument described in this paper utilizes the fundamental and second-harmonic output from an injection seeded Nd:YAG laser as the laser transmitter. The light scattered in the atmosphere is collected using a commercial Schmidt-Cassegrain telescope with the optical receiver train first splitting the fundamental and second-harmonic return signal with the fundament light monitored using an avalanche photodiode. The second-harmonic return signal is mode matched into a tunable confocal Fabry-Perot (CFP) interferometer with a free spectral range of 7.5 GHz and a finesse of 50.7 (312) at 532 nm (1064 nm) placed in the optical receiver for spectrally filtering the molecular and aerosol return signals. The light transmitted through the CFP is used to monitor the aerosol return signal while the light reflected from the CFP is used to monitor the molecular return signal. Data collected with the HSRL are presented and inversion results are compared to a co-located solar radiometer, demonstrating the successful operation of the instrument. The CFP-based filtering technique successfully employed by this HSRL instrument is easily portable to other arbitrary wavelengths, thus allowing for the future development of multiwavelength HSRL instruments.

Documentation of a Conduit Flow Process (CFP) for MODFLOW-2005

USGS Publications Warehouse

Shoemaker, W. Barclay; Kuniansky, Eve L.; Birk, Steffen; Bauer, Sebastian; Swain, Eric D.

2007-01-01

This report documents the Conduit Flow Process (CFP) for the modular finite-difference ground-water flow model, MODFLOW-2005. The CFP has the ability to simulate turbulent ground-water flow conditions by: (1) coupling the traditional ground-water flow equation with formulations for a discrete network of cylindrical pipes (Mode 1), (2) inserting a high-conductivity flow layer that can switch between laminar and turbulent flow (Mode 2), or (3) simultaneously coupling a discrete pipe network while inserting a high-conductivity flow layer that can switch between laminar and turbulent flow (Mode 3). Conduit flow pipes (Mode 1) may represent dissolution or biological burrowing features in carbonate aquifers, voids in fractured rock, and (or) lava tubes in basaltic aquifers and can be fully or partially saturated under laminar or turbulent flow conditions. Preferential flow layers (Mode 2) may represent: (1) a porous media where turbulent flow is suspected to occur under the observed hydraulic gradients; (2) a single secondary porosity subsurface feature, such as a well-defined laterally extensive underground cave; or (3) a horizontal preferential flow layer consisting of many interconnected voids. In this second case, the input data are effective parameters, such as a very high hydraulic conductivity, representing multiple features. Data preparation is more complex for CFP Mode 1 (CFPM1) than for CFP Mode 2 (CFPM2). Specifically for CFPM1, conduit pipe locations, lengths, diameters, tortuosity, internal roughness, critical Reynolds numbers (NRe), and exchange conductances are required. CFPM1, however, solves the pipe network equations in a matrix that is independent of the porous media equation matrix, which may mitigate numerical instability associated with solution of dual flow components within the same matrix. CFPM2 requires less hydraulic information and knowledge about the specific location and hydraulic properties of conduits, and turbulent flow is approximated by modifying horizontal conductances assembled by the Block-Centered Flow (BCF), Layer-Property Flow (LPF), or Hydrogeologic-Unit Flow Packages (HUF) of MODFLOW-2005. For both conduit flow pipes (CFPM1) and preferential flow layers (CFPM2), critical Reynolds numbers are used to determine if flow is laminar or turbulent. Due to conservation of momentum, flow in a laminar state tends to remain laminar and flow in a turbulent state tends to remain turbulent. This delayed transition between laminar and turbulent flow is introduced in the CFP, which provides an additional benefit of facilitating convergence of the computer algorithm during iterations of transient simulations. Specifically, the user can specify a higher critical Reynolds number to determine when laminar flow within a pipe converts to turbulent flow, and a lower critical Reynolds number for determining when a pipe with turbulent flow switches to laminar flow. With CFPM1, the Hagen-Poiseuille equation is used for laminar flow conditions and the Darcy-Weisbach equation is applied to turbulent flow conditions. With CFPM2, turbulent flow is approximated by reducing the laminar hydraulic conductivity by a nonlinear function of the Reynolds number, once the critical head difference is exceeded. This adjustment approximates the reductions in mean velocity under turbulent ground-water flow conditions.

Alternative Fuels Data Center: Propane Mowers Help National Park Cut

Science.gov Websites

vehicle fleet will run on alternative fuels by 2014. With the increased number of AFVs, the park is said that the project has had a positive influence on park staff, other NPS units, local municipality national park in Texas to meet the requirements of the Climate Friendly Parks (CFP) program. CFP is one

Color Fundus Photography, Optical Coherence Tomography, and Fluorescein Angiography in Diagnosing Polypoidal Choroidal Vasculopathy.

PubMed

Chaikitmongkol, Voraporn; Khunsongkiet, Preeyanuch; Patikulsila, Direk; Ratanasukon, Mansing; Watanachai, Nawat; Jumroendararasame, Chaisiri; Mayerle, Catherine B; Han, Ian C; Chen, Connie J; Winaikosol, Pawara; Dejkriengkraikul, Chutikarn; Choovuthayakorn, Janejit; Kunavisarut, Paradee; Bressler, Neil M

2018-05-10

To determine sensitivity and specificity of polypoidal choroidal vasculopathy (PCV) diagnosis using color fundus photography (CFP), optical coherence tomography (OCT) and fundus fluorescein angiography (FFA) without indocyanine-green angiography (ICGA). Validity analysis. Treatment-naïve eyes with serous/serosanguinous maculopathy undergoing CFP, OCT, FFA and ICGA imaging before treatment at a university-hospital in Thailand (January 2013 to June 2015) were identified. Images of each subject were categorized into 4 sets (set A: CFP; set B: CFP+OCT; set C: CFP+FFA; set D: CFP+OCT+FFA). Six graders, 3 from Thailand (PCV endemic area) and 3 from U.S. (non-endemic area), individually reviewed each set (without ICG), and determined if the presumed diagnosis was PCV. In parallel, 2 other graders confirmed if each case had PCV or not using EVEREST criteria (including ICGA). Sensitivity and specificity of a PCV diagnosis with each set (without ICGA) were analyzed compared with diagnoses including ICGA. Of 119 study eyes (113 subjects, 57% male, mean age±SD 59.9±13.8), definite PCV diagnosis was 40.3%. Sensitivity of sets A, B, C, D: 0.63 (95%CI: 0.47-0.76), 0.83 (95%CI: 0.69-0.92), 0.54 (95%CI: 0.39-0.68), 0.67 (95%CI: 0.51-0.79). Specificity were 0.93 (95% CI: 0.84-0.97), 0.83 (95%CI: 0.72-0.91), 0.97 (95%CI: 0.89-0.99), 0.92 (95%CI: 0.82-0.97). Accuracies: 0.81 (95%CI: 0.73-0.88), 0.83 (95%CI: 0.76-0.90), 0.79 (95%CI: 0.73-0.87), 0.82 (95%CI: 0.74-0.88). Discrepancies between Thai and US graders existed through sets A, C, and D. These data suggest without ICGA, fundus photography combined with OCT provides high sensitivity and high specificity to diagnosis PCV; adding FFA does not improve accuracy. Copyright © 2018 Elsevier Inc. All rights reserved.

Development of a Nanostructured α-MnO2/Carbon Paper Composite for Removal of Ni2+ / Mn2+ ions by Electrosorption.

PubMed

Li, Pengju; Gui, Yang; Blackwood, Daniel John

2018-05-22

Toxic metal ions, such as Ni2+ and Mn2+, in industrial waste streams are non-biodegradable and can cause damage to the human body. Electrochemical cleaning techniques are attractive as they offer more control and produce less sludge than chemical / biological approaches without the high pressures needed for membranes. Here nanoneedle structured α-MnO2/carbon fiber paper (CFP) composites were synthesized by a hydrothermal approach and used as electrodes for combined electro-adsorption and capacitive deionization removal of nickel and manganese ions from pseudo industrial waste streams. The specific performance of α-MnO2/CFP (16.4 mg Ni2+ per gram of active material) not only shows a great improve in comparison with its original CFP substrate (0.034 Ni2+ mg per gram), but is over six times that of activated carbon (2.5 mg Ni2+ per gram). The high performance of α-MnO2/CFP composite is attributed to its high surface area, desirable mesoporosity and pore size distribution that permits the further access of ions, and the property as a pseudocapacitor, which contributes to a more efficient electron/charge transfer in the faradic process. Unfortunately, it was also found that some Mn2+ ions are released due to partial reduction of the MnO2 when operated as a negative electrode. For the removal of Mn2+ ions an asymmetric arrangement, consisting of a MnO2/CFP positive electrode and an activated carbon negative electrode was employed. This arrangement reduced the Mn2+ concentration from 100 ppm to less than 2 ppm, a vast improvement over a systematical two activated carbon electrodes system that could only reach 42 ppm under the same conditions. It was also observed that as long as the MnO2/CFP composite was maintained as a positive electrode it was completely stable. The technique was able to reduce both Ni2+ and Mn2+ ions to well below the 10 ppm requirement for discharge into public sewers in Singapore.

Pilot study of home-based delivery of HIV testing and counseling and contraceptive services to couples in Malawi.

PubMed

Becker, Stan; Taulo, Frank O; Hindin, Michelle J; Chipeta, Effie K; Loll, Dana; Tsui, Amy

2014-12-20

HIV counseling and testing for couples is an important component of HIV prevention strategies, particularly in Sub Saharan Africa. The purpose of this pilot study is to estimate the uptake of couple HIV counseling and testing (CHCT) and couple family planning (CFP) services in a single home visit in peri-urban Malawi and to assess related factors. This study involved offering CHCT and CFP services to couples in their homes; 180 couples were sampled from households in a peri-urban area of Blantyre. Baseline data were collected from both partners and follow-up data were collected one week later. A pair of male and female counselors approached each partner separately about HIV testing and counseling and contraceptive services and then, if both consented, CHCT and CFP services (pills, condoms and referrals for other methods) were given. Bivariate and multivariate logistic regression analyses were done to examine the relationship between individual partner characteristics and acceptance of the services. Selected behaviors reported pre- and post-intervention, particularly couple reports on contraceptive use and condom use at last sex, were also tested for differences. 89% of couples accepted at least one of the services (58% CHCT-only, 29% CHCT + CFP, 2% CFP-only). Among women, prior testing experience (p < 0.05), parity (p < 0.01), and emotional closeness to partner (p < 0.01) had significant bivariate associations with acceptance of at least one service. Reported condom use at last sex increased from 6% to 25% among couples receiving any intervention. First-ever HIV testing was delivered to 25 women and 69 men, resulting, respectively, in 4 and 11 newly detected infections. Home-based CHCT and CFP were very successful in this pilot study with high proportions of previously untested husbands and wives accepting CHCT and there were virtually no negative outcomes within one week. This study supports the need for further research and testing of home- and couple-based approaches to expand access to HCT and contraceptive services to prevent the undesired consequences of sexually transmitted infection and unintended pregnancy via unprotected sex.

Intrinsically incompatible crystal (ligand) field parameter sets for transition ions at orthorhombic and lower symmetry sites in crystals and their implications

NASA Astrophysics Data System (ADS)

Rudowicz, C.; Gnutek, P.

2010-01-01

Central quantities in spectroscopy and magnetism of transition ions in crystals are crystal (ligand) field parameters (CFPs). For orthorhombic, monoclinic, and triclinic site symmetry CF analysis is prone to misinterpretations due to large number of CFPs and existence of correlated sets of alternative CFPs. In this review, we elucidate the intrinsic features of orthorhombic and lower symmetry CFPs and their implications. The alternative CFP sets, which yield identical energy levels, belong to different regions of CF parameter space and hence are intrinsically incompatible. Only their ‘images’ representing CFP sets expressed in the same region of CF parameter space may be directly compared. Implications of these features for fitting procedures and meaning of fitted CFPs are categorized into negative: pitfalls and positive: blessings. As a case study, the CFP sets for Tm 3+ ions in KLu(WO 4) 2 are analysed and shown to be intrinsically incompatible. Inadvertent, so meaningless, comparisons of incompatible CFP sets result in various pitfalls, e.g., controversial claims about the values of CFPs obtained by other researchers as well as incorrect structural conclusions or faulty systematics of CF parameters across rare-earth ion series based on relative magnitudes of incompatible CFPs. Such pitfalls bear on interpretation of, e.g., optical spectroscopy, inelastic neutron scattering, and magnetic susceptibility data. An extensive survey of pertinent literature was carried out to assess recognition of compatibility problems. Great portion of available orthorhombic and lower symmetry CFP sets are found intrinsically incompatible, yet these problems and their implications appear barely recognized. The considerable extent and consequences of pitfalls revealed by our survey call for concerted remedial actions of researchers. A general approach based on the rhombicity ratio standardization may solve compatibility problems. Wider utilization of alternative CFP sets in the multiple correlated fitting techniques may improve reliability ( blessing) of fitted CFPs. This review may be of interest to a broad range of researchers from condensed matter physicists to physical chemists working on, e.g., high temperature superconductors, luminescent, optoelectronic, laser, and magnetic materials.

Differences in the volume of pharmaceutical advertisements between print general medical journals.

PubMed

Gettings, Jennifer; O'Neill, Braden; Chokshi, Dave A; Colbert, James A; Gill, Peter; Lebovic, Gerald; Lexchin, Joel; Persaud, Navindra

2014-01-01

Pharmaceutical advertisements have been argued to provide revenue that medical journals require but they are intended to alter prescribing behaviour and they are known to include low quality information. We determined whether a difference exists in the current level of pharmaceutical advertising in print general medical journals, and we estimated the revenue generated from print pharmaceutical advertising. Six print general medical journals in Canada, the United States, and the United Kingdom were sampled between 2007 and 2012. The number of advertisements and other journal content in selected issues of the Canadian Medical Association Journal (CMAJ), Canadian Family Physician (CFP), Journal of the American Medical Association (JAMA), New England Journal of Medicine (NEJM), British Medical Journal (BMJ), and Lancet were determined. Revenue gained from pharmaceutical advertising was estimated using each journal's 2013 advertising price list. The two Canadian journals sampled (CMAJ, CFP) contained five times more advertisements than the two American journals (JAMA, NEJM), and two British journals (BMJ, Lancet) (p<0.0001). The estimated annual revenue from pharmaceutical advertisements ranged from £0.025 million (for Lancet) to £3.8 million (for JAMA). The cost savings due to revenue from pharmaceutical advertising to each individual subscriber ranged from £0.02 (for Lancet) to £3.56 (for CFP) per issue. The volume of pharmaceutical advertisements differs between general medical journals, with the two Canadian journals sampled containing the most advertisements. International and temporal variations suggest that there is an opportunity for all general medical journals to reduce the number of pharmaceutical advertisements, explore other sources of revenue, and increase transparency regarding sources of revenue.

A study of IEEE 802.15.4 security framework for wireless body area networks.

PubMed

Saleem, Shahnaz; Ullah, Sana; Kwak, Kyung Sup

2011-01-01

A Wireless Body Area Network (WBAN) is a collection of low-power and lightweight wireless sensor nodes that are used to monitor the human body functions and the surrounding environment. It supports a number of innovative and interesting applications, including ubiquitous healthcare and Consumer Electronics (CE) applications. Since WBAN nodes are used to collect sensitive (life-critical) information and may operate in hostile environments, they require strict security mechanisms to prevent malicious interaction with the system. In this paper, we first highlight major security requirements and Denial of Service (DoS) attacks in WBAN at Physical, Medium Access Control (MAC), Network, and Transport layers. Then we discuss the IEEE 802.15.4 security framework and identify the security vulnerabilities and major attacks in the context of WBAN. Different types of attacks on the Contention Access Period (CAP) and Contention Free Period (CFP) parts of the superframe are analyzed and discussed. It is observed that a smart attacker can successfully corrupt an increasing number of GTS slots in the CFP period and can considerably affect the Quality of Service (QoS) in WBAN (since most of the data is carried in CFP period). As we increase the number of smart attackers the corrupted GTS slots are eventually increased, which prevents the legitimate nodes to utilize the bandwidth efficiently. This means that the direct adaptation of IEEE 802.15.4 security framework for WBAN is not totally secure for certain WBAN applications. New solutions are required to integrate high level security in WBAN.

A Study of IEEE 802.15.4 Security Framework for Wireless Body Area Networks

PubMed Central

Saleem, Shahnaz; Ullah, Sana; Kwak, Kyung Sup

2011-01-01

A Wireless Body Area Network (WBAN) is a collection of low-power and lightweight wireless sensor nodes that are used to monitor the human body functions and the surrounding environment. It supports a number of innovative and interesting applications, including ubiquitous healthcare and Consumer Electronics (CE) applications. Since WBAN nodes are used to collect sensitive (life-critical) information and may operate in hostile environments, they require strict security mechanisms to prevent malicious interaction with the system. In this paper, we first highlight major security requirements and Denial of Service (DoS) attacks in WBAN at Physical, Medium Access Control (MAC), Network, and Transport layers. Then we discuss the IEEE 802.15.4 security framework and identify the security vulnerabilities and major attacks in the context of WBAN. Different types of attacks on the Contention Access Period (CAP) and Contention Free Period (CFP) parts of the superframe are analyzed and discussed. It is observed that a smart attacker can successfully corrupt an increasing number of GTS slots in the CFP period and can considerably affect the Quality of Service (QoS) in WBAN (since most of the data is carried in CFP period). As we increase the number of smart attackers the corrupted GTS slots are eventually increased, which prevents the legitimate nodes to utilize the bandwidth efficiently. This means that the direct adaptation of IEEE 802.15.4 security framework for WBAN is not totally secure for certain WBAN applications. New solutions are required to integrate high level security in WBAN. PMID:22319358

Direct interaction of Plin2 with lipids on the surface of lipid droplets: a live cell FRET analysis

PubMed Central

McIntosh, Avery L.; Senthivinayagam, Subramanian; Moon, Kenneth C.; Gupta, Shipra; Lwande, Joel S.; Murphy, Cameron C.; Storey, Stephen M.

2012-01-01

Despite increasing awareness of the health risks associated with excess lipid storage in cells and tissues, knowledge of events governing lipid exchange at the surface of lipid droplets remains unclear. To address this issue, fluorescence resonance energy transfer (FRET) was performed to examine live cell interactions of Plin2 with lipids involved in maintaining lipid droplet structure and function. FRET efficiencies (E) between CFP-labeled Plin2 and fluorescently labeled phosphatidylcholine, sphingomyelin, stearic acid, and cholesterol were quantitated on a pixel-by-pixel basis to generate FRET image maps that specified areas with high E (>60%) in lipid droplets. The mean E and the distance R between the probes indicated a high yield of energy transfer and demonstrated molecular distances on the order of 44–57 Å, in keeping with direct molecular contact. In contrast, FRET between CFP-Plin2 and Nile red was not detected, indicating that the CFP-Plin2/Nile red interaction was beyond FRET proximity (>100 Å). An examination of the effect of Plin2 on cellular metabolism revealed that triacylglycerol, fatty acid, and cholesteryl ester content increased while diacylglycerol remained constant in CFP-Plin2-overexpressing cells. Total phospholipids also increased, reflecting increased phosphatidylcholine and sphingomyelin. Consistent with these results, expression levels of enzymes involved in triacylglycerol, cholesteryl ester, and phospholipid synthesis were significantly upregulated in CFP-Plin2-expressing cells while those associated with lipolysis either decreased or were unaffected. Taken together, these data show for the first time that Plin2 interacts directly with lipids on the surface of lipid droplets and influences levels of key enzymes and lipids involved in maintaining lipid droplet structure and function. PMID:22744009

Effects of intra-aortic balloon pump versus centrifugal pump on myocardial energetics and systemic circulation in a porcine model of rapidly worsening acute heart failure.

PubMed

Ntalianis, Argyrios S; Drakos, Stavros G; Charitos, Christos; Dolou, Paraskevi; Pierrakos, Charalampos N; Terrovitis, John V; Papaioannou, Theodoros; Charitos, Efstratios; Nanas, John N

2008-01-01

The present experimental study compared the effectiveness of counterpulsation provided by the intra-aortic balloon pump (IABP) versus that of a nonpulsatile, radial-flow centrifugal pump (CFP) in rapidly worsening acute heart failure (HF). Eighteen pigs were included in the study. After the induction of acute moderate HF, circulatory support was randomly provided with either the IABP or CFP. No significant change in cardiac output (CO) and mean aortic pressure (MAP) was observed with either pump. The IABP caused a significantly greater decrease than the CFP in 1) double product (13.138 +/- 2.476 mm Hg/min vs. 14.217 +/- 2.673 mm Hg/min, p = 0.023), 2) left ventricular systolic pressure (LVSP, 100 +/- 8 mm Hg vs. 106 +/- 10 mm Hg, p = 0.046), and 3) end-diastolic aortic pressure (EDAP, 70 +/- 6 mm Hg vs. 86 +/- 6 mm Hg, p = 0.000). The effects of both pumps on total tension time index and LAD flow were similar. After the induction of severe HF, the IABP had its main effects on afterload and decreased LVSP from 88 +/- 6 mm Hg to 78 +/- 9 mm Hg, (p = 0.008), and EDAP from 57 +/- 9 mm Hg to 49 +/- 14 mm Hg, (p = 0.044), whereas the CFP exerted its effects mainly on preload, lowering LV end-diastolic pressure from 19 +/- 5 mm Hg to 11 +/- 4 mm Hg, (p = 0.002). CO and MAP were similarly increased by both assist systems. The IABP (by lowering afterload) and CFP (by lowering preload) both offered significant mechanical support in acute HF. However, afterload reduction offered principally by the IABP seems preferable for the recovery of the acutely failing heart.

NETosing Neutrophils Activate Complement Both on Their Own NETs and Bacteria via Alternative and Non-alternative Pathways

PubMed Central

Yuen, Joshua; Pluthero, Fred G.; Douda, David N.; Riedl, Magdalena; Cherry, Ahmed; Ulanova, Marina; Kahr, Walter H. A.; Palaniyar, Nades; Licht, Christoph

2016-01-01

Neutrophils deposit antimicrobial proteins, such as myeloperoxidase and proteases on chromatin, which they release as neutrophil extracellular traps (NETs). Neutrophils also carry key components of the complement alternative pathway (AP) such as properdin or complement factor P (CFP), complement factor B (CFB), and C3. However, the contribution of these complement components and complement activation during NET formation in the presence and absence of bacteria is poorly understood. We studied complement activation on NETs and a Gram-negative opportunistic bacterial pathogen Pseudomonas aeruginosa (PA01, PAKwt, and PAKgfp). Here, we show that anaphylatoxin C5a, formyl-methionyl-leucyl-phenylalanine (fMLP) and phorbol myristate acetate (PMA), which activates NADPH oxidase, induce the release of CFP, CFB, and C3 from neutrophils. In response to PMA or P. aeruginosa, neutrophils secrete CFP, deposit it on NETs and bacteria, and induce the formation of terminal complement complexes (C5b–9). A blocking anti-CFP antibody inhibited AP-mediated but not non-AP-mediated complement activation on NETs and P. aeruginosa. Therefore, NET-mediated complement activation occurs via both AP- and non AP-based mechanisms, and AP-mediated complement activation during NETosis is dependent on CFP. These findings suggest that neutrophils could use their “AP tool kit” to readily activate complement on NETs and Gram-negative bacteria, such as P. aeruginosa, whereas additional components present in the serum help to fix non-AP-mediated complement both on NETs and bacteria. This unique mechanism may play important roles in host defense and help to explain specific roles of complement activation in NET-related diseases. PMID:27148258

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

Ferrell, Jack R; Ware, Anne E

Two catalytic fast pyrolysis (CFP) oils (bottom/heavy fraction) were analyzed in various solvents that are used in common analytical methods (nuclear magnetic resonance - NMR, gas chromatography - GC, gel permeation chromatography - GPC, thermogravimetric analysis - TGA) for oil characterization and speciation. A more accurate analysis of the CFP oils can be obtained by identification and exploitation of solvent miscibility characteristics. Acetone and tetrahydrofuran can be used to completely solubilize CFP oils for analysis by GC and tetrahydrofuran can be used for traditional organic GPC analysis of the oils. DMSO-d6 can be used to solubilize CFP oils for analysismore » by 13C NMR. The fractionation of oils into solvents that did not completely solubilize the whole oils showed that miscibility can be related to the oil properties. This allows for solvent selection based on physico-chemical properties of the oils. However, based on semi-quantitative comparisons of the GC chromatograms, the organic solvent fractionation schemes did not speciate the oils based on specific analyte type. On the other hand, chlorinated solvents did fractionate the oils based on analyte size to a certain degree. Unfortunately, like raw pyrolysis oil, the matrix of the CFP oils is complicated and is not amenable to simple liquid-liquid extraction (LLE) or solvent fractionation to separate the oils based on the chemical and/or physical properties of individual components. For reliable analyses, for each analytical method used, it is critical that the bio-oil sample is both completely soluble and also not likely to react with the chosen solvent. The adoption of the standardized solvent selection protocols presented here will allow for greater reproducibility of analysis across different users and facilities.« less

Recombinant ESAT-6-CFP10 Fusion Protein Induction of Th1/Th2 Cytokines and FoxP3 Expressing Treg Cells in Pulmonary TB.

PubMed

Jackson-Sillah, Dolly; Cliff, Jacqueline M; Mensah, Gloria Ivy; Dickson, Emmanuel; Sowah, Sandra; Tetteh, John K A; Addo, Kwasi K; Ottenhoff, Tom H M; Bothamley, Graham; Dockrell, Hazel M

2013-01-01

Early secretory antigenic target 6 (ESAT-6) and culture filtrate protein 10 (CFP-10) are Mycobacterium tuberculosis (Mtb)-specific antigens that are secreted by actively metabolising bacteria and contribute to the virulence of the bacteria. Their ability to induce Treg and Th2 responses, particularly during the first two weeks of treatment, has not been comprehensively examined to date. The purpose of this work was to characterise Th1, Th2 and Treg responses to rESAT-6-CFP10 fusion protein in TB patients before and during the intensive phase of treatment and in healthy M.bovis BCG vaccinated donors. Forty-six newly diagnosed, HIV-negative, smear-positive pulmonary TB patients and 20 healthy donors were recruited in the UK and Ghana. Their peripheral blood mononuclear cells (PBMC) were used in ex vivo ELISPOT and in vitro cultures to identify immunological parameters of interest. The study confirmed that protective immune responses to rESAT-6-CFP10 are impaired in active TB but improved during treatment: circulating antigen-specific IL-4-producing T-cells were increased in untreated TB but declined by two weeks of treatment while the circulating antigen-specific IFN-γ producing T cells which showed a transient rise at one week of treatment, persisted at baseline levels at two months of treatment. In vitro T cell proliferation and IFN-γ production were reduced, while IL-4 and CD4(+)FoxP3(+)CD25(hi) cell expression were increased in response to rESAT-6-CFP10 fusion protein in untreated TB. These responses were reversed during early treatment of TB. These observations support further investigations into the possible utility of these parameters as markers of active disease and favourable treatment outcomes.

Surprisingly high specificity of the PPD skin test for M. tuberculosis infection from recent exposure in The Gambia.

PubMed

Hill, Philip C; Brookes, Roger H; Fox, Annette; Jackson-Sillah, Dolly; Lugos, Moses D; Jeffries, David J; Donkor, Simon A; Adegbola, Richard A; McAdam, Keith P W J

2006-12-20

Options for intervention against Mycobacterium tuberculosis infection are limited by the diagnostic tools available. The Purified Protein Derivative (PPD) skin test is thought to be non-specific, especially in tropical settings. We compared the PPD skin test with an ELISPOT test in The Gambia. Household contacts over six months of age of sputum smear positive TB cases and community controls were recruited. They underwent a PPD skin test and an ELISPOT test for the T cell response to PPD and ESAT-6/CFP10 antigens. Responsiveness to M. tuberculosis exposure was analysed according to sleeping proximity to an index case using logistic regression. 615 household contacts and 105 community controls were recruited. All three tests assessed increased significantly in positivity with increasing M. tuberculosis exposure, the PPD skin test most dramatically (OR 15.7; 95% CI 6.6-35.3). While the PPD skin test positivity continued to trend downwards in the community with increasing distance from a known case (61.9% to 14.3%), the PPD and ESAT-6/CFP-10 ELISPOT positivity did not. The PPD skin test was more in agreement with ESAT-6/CFP-10 ELISPOT (75%, p = 0.01) than the PPD ELISPOT (53%, p<0.0001). With increasing M. tuberculosis exposure, the proportion of ESAT-6/CFP-10 positive contacts who were PPD skin test positive increased (p<0.0001), and the proportion of ESAT-6/CFP-10 negative contacts that were PPD skin test negative decreased (p<0.0001); the converse did not occur. The PPD skin test has surprisingly high specificity for M. tuberculosis infection from recent exposure in The Gambia. In this setting, anti-tuberculous prophylaxis in PPD skin test positive individuals should be revisited.

Detection of circulating Mycobacterium tuberculosis-specific DNA by droplet digital PCR for vaccine evaluation in challenged monkeys and TB diagnosis.

PubMed

Song, Neng; Tan, Yang; Zhang, Lingyun; Luo, Wei; Guan, Qing; Yan, Ming-Zhe; Zuo, Ruiqi; Liu, Weixiang; Luo, Feng-Ling; Zhang, Xiao-Lian

2018-04-24

Mycobacterium tuberculosis (M. tb) is emerging as a more serious pathogen due to the increased multidrug-resistant TB and co-infection of human immunodeficiency virus (HIV). The development of an effective and sensitive detection method is urgently needed for bacterial load evaluation in vaccine development, early TB diagnosis, and TB treatment. Droplet digital polymerase chain reaction (ddPCR) is a newly developed sensitive PCR method for the absolute quantification of nucleic acid concentrations. Here, we used ddPCR to quantify the circulating virulent M. tb-specific CFP10 (10-kDa culture filtrate protein, Rv3874) and Rv1768 DNA copy numbers in the blood samples from Bacille Calmette-Guerin (BCG)-vaccinated and/or virulent M. tb H37Rv-challenged rhesus monkeys. We found that ddPCR was more sensitive compared to real-time fluorescence quantitative PCR (qPCR), as the detection limits of CFP10 were 1.2 copies/μl for ddPCR, but 15.8 copies/μl for qPCR. We demonstrated that ddPCR could detect CFP10 and Rv1768 DNA after 3 weeks of infection and at least two weeks earlier than qPCR in M.tb H37Rv-challenged rhesus monkey models. DdPCR could also successfully quantify CFP10 and Rv1768 DNA copy numbers in clinical TB patients' blood samples (active pulmonary TB, extrapulmonary TB (EPTB), and infant TB). To our knowledge, this study is the first to demonstrate that ddPCR is an effective and sensitive method of measuring the circulating CFP10 and Rv1768 DNA for vaccine development, bacterial load evaluation in vivo, and early TB (including EPTB and infant TB) diagnosis as well.

Ciguatera fish poisoning in la Habana, Cuba: a study of local social-ecological resilience.

PubMed

Morrison, Karen; Aguiar Prieto, Pablo; Castro Domínguez, Arnaldo; Waltner-Toews, David; Fitzgibbon, John

2008-09-01

Following the collapse of the Cuban economy in the early 1990s, epidemiologists in the Cuban Ministry of Health noticed dramatic increases in reported outbreaks of ciguatera fish poisoning (CFP) in some coastal communities. This article summarizes the results of a comparative case study which applied an ecosystem approach to human health to investigate this issue. Situated learning and complexity theories were used to interpret the results of the investigation. CFP outbreaks are influenced by a complex set of interactions between ecological and socioeconomic processes. This study found that the level of organization of the local sports fishing community and the degree of degradation of the local nearshore marine ecosystem appear to be key factors influencing the diverging levels of CFP outbreaks recorded in the 1990s in the communities studied.

FRET-based binding assay between a fluorescent cAMP analogue and a cyclic nucleotide-binding domain tagged with a CFP.

PubMed

Romero, Francisco; Santana-Calvo, Carmen; Sánchez-Guevara, Yoloxochitl; Nishigaki, Takuya

2017-09-01

The cyclic nucleotide-binding domain (CNBD) functions as a regulatory domain of many proteins involved in cyclic nucleotide signalling. We developed a straightforward and reliable binding assay based on intermolecular fluorescence resonance energy transfer (FRET) between an adenosine-3', 5'-cyclic monophosphate analogue labelled with fluorescein and a recombinant CNBD of human EPAC1 tagged with a cyan fluorescence protein (CFP). The high FRET efficiency of this method (~ 80%) allowed us to perform several types of binding experiments with nanomolar range of sample using conventional equipment. In addition, the CFP tag on the CNBD enabled us to perform a specific binding experiment using an unpurified protein. Considering these advantages, this technique is useful to study poorly characterized CNBDs. © 2017 Federation of European Biochemical Societies.

Estimating Carbon Flux Phenology with Satellite-Derived Land Surface Phenology and Climate Drivers for Different Biomes: A Synthesis of AmeriFlux Observations

PubMed Central

Zhu, Wenquan; Chen, Guangsheng; Jiang, Nan; Liu, Jianhong; Mou, Minjie

2013-01-01

Carbon Flux Phenology (CFP) can affect the interannual variation in Net Ecosystem Exchange (NEE) of carbon between terrestrial ecosystems and the atmosphere. In this study, we proposed a methodology to estimate CFP metrics with satellite-derived Land Surface Phenology (LSP) metrics and climate drivers for 4 biomes (i.e., deciduous broadleaf forest, evergreen needleleaf forest, grasslands and croplands), using 159 site-years of NEE and climate data from 32 AmeriFlux sites and MODIS vegetation index time-series data. LSP metrics combined with optimal climate drivers can explain the variability in Start of Carbon Uptake (SCU) by more than 70% and End of Carbon Uptake (ECU) by more than 60%. The Root Mean Square Error (RMSE) of the estimations was within 8.5 days for both SCU and ECU. The estimation performance for this methodology was primarily dependent on the optimal combination of the LSP retrieval methods, the explanatory climate drivers, the biome types, and the specific CFP metric. This methodology has a potential for allowing extrapolation of CFP metrics for biomes with a distinct and detectable seasonal cycle over large areas, based on synoptic multi-temporal optical satellite data and climate data. PMID:24386441

Ability of certain plant extracts traditionally used to treat ciguatera fish poisoning to inhibit nitric oxide production in RAW 264.7 macrophages.

PubMed

Kumar-Roiné, Shilpa; Matsui, Mariko; Reybier, Karine; Darius, Hélène Taiana; Chinain, Mireille; Pauillac, Serge; Laurent, Dominique

2009-06-25

Ciguatera fish poisoning (CFP) is an intertropical ichthyosarcotoxism that manifests in complex assortment of symptoms in humans. Ciguatoxins (CTXs), issued from Gambierdicus spp., are causative agents of this intoxication. We have recently demonstrated that a Pacific CTX (P-CTX-1B) strongly modulated iNOS expression, leading to overproduction of nitric oxide (NO) in RAW 264.7 murine macrophage cells. NO produced in large amounts is involved in a wide range of pathophysiological processes. Many traditional remedies are commonly used in the Pacific against CFP. In this context, bioassay-guided screening was carried out to study NO inhibiting capacity of 28 selected plant extracts. We prepared aqueous extracts of plants used in New Caledonia in the treatment of CFP and screened their NO inhibitory activity in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. Among 28 plants tested, Euphorbia hirta (Euphorbiaceae), Syzygium malaccense (Myrtaceae), Schinus terebenthifolius (Anacardiaceae), Punica granatum (Punicaceae), Cerbera manghas (Apocynaceae), Vitex trifolia (Labiateae) and Ximenia americana (Olacaceae) showed inhibitory activity, validating their use as traditional remedies in CFP, and the potential for use in the treatment of conditions accompanied by NO overproduction. These plants are promising candidates for further screening of their active compounds through activity-guided fractionation.

Cluster of ciguatera fish poisoning--North Carolina, 2007.

PubMed

2009-03-27

Ciguatera fish poisoning (CFP) is a distinctive type of foodborne disease that results from eating predatory ocean fish contaminated with ciguatoxins. As many as 50,000 cases are reported worldwide annually, and the condition is endemic in tropical and subtropical regions of the Pacific basin, Indian Ocean, and Caribbean. In the United States, 5--70 cases per 10,000 persons are estimated to occur yearly in ciguatera-endemic states and territories. CFP can cause gastrointestinal symptoms (nausea, vomiting, abdominal cramps, or diarrhea) within a few hours of eating contaminated fish. Neurologic symptoms, with or without gastrointestinal disturbance, can include fatigue, muscle pain, itching, tingling, and (most characteristically) reversal of hot and cold sensation. This report describes a cluster of nine cases of CFP that occurred in North Carolina in June 2007. Among the nine patients, six experienced reversal of hot and cold sensations, five had neurologic symptoms only, and overall symptoms persisted for more than 6 months in three patients. Among seven patients who were sexually active, six patients also complained of painful intercourse. This report highlights the potential risks of eating contaminated ocean fish. Local and state health departments can train emergency and urgent care physicians in the recognition of CFP and make them aware that symptoms can persist for months to years.

Estimating Carbon Flux Phenology with Satellite-Derived Land Surface Phenology and Climate Drivers for Different Biomes: A Synthesis of AmeriFlux Observations

DOE PAGES

Zhu, Wenquan; Chen, Guangsheng; Jiang, Nan; ...

2013-12-27

Carbon Flux Phenology (CFP) can affect the interannual variation in Net Ecosystem Exchange (NEE) of carbon between terrestrial ecosystems and the atmosphere. In this paper, we proposed a methodology to estimate CFP metrics with satellite-derived Land Surface Phenology (LSP) metrics and climate drivers for 4 biomes (i.e., deciduous broadleaf forest, evergreen needleleaf forest, grasslands and croplands), using 159 site-years of NEE and climate data from 32 AmeriFlux sites and MODIS vegetation index time-series data. LSP metrics combined with optimal climate drivers can explain the variability in Start of Carbon Uptake (SCU) by more than 70% and End of Carbon Uptakemore » (ECU) by more than 60%. The Root Mean Square Error (RMSE) of the estimations was within 8.5 days for both SCU and ECU. The estimation performance for this methodology was primarily dependent on the optimal combination of the LSP retrieval methods, the explanatory climate drivers, the biome types, and the specific CFP metric. In conclusion, this methodology has a potential for allowing extrapolation of CFP metrics for biomes with a distinct and detectable seasonal cycle over large areas, based on synoptic multi-temporal optical satellite data and climate data.« less

Microenvironments and Signaling Pathways Regulating Early Dissemination, Dormancy, and Metastasis

DTIC Science & Technology

2015-09-01

late disseminated tumor cells. Progress has also been made in devising a sorting protocol to isolate CFP+/HER2+ DTCs from lungs and CTCs in the MMTV...early dissemination of pre- malignant MECs. We further hypothesize that after extravasation at S-TMEM, in order to exit dormancy a stable S-TMEM...mentioned in section SA1.2, and the optimization of these protocols is underway. Aim 2.1 requires that we validate the components of the TMEM in the

Ciguatera fish poisoning.

PubMed

Armstrong, Patrick; Murray, Peter; Nesdale, Annette; Peckler, Brad

2016-10-28

Ciguatera fish poisoning (CFP) is the most common cause of seafood-toxin poisoning in the world and is most prevalent in tropical and subtropical areas. It causes gastroenteritis but also myriad neurological and cardiovascular symptoms. We present a cluster of CFP that occurred in Wellington Hospital, New Zealand. It resulted in three patients with life threatening cardiotoxicity and a fourth case with severe gastro-intestinal symptoms. The epidemiology, clinical manifestations, diagnosis, treatment and public health issues are discussed.

Enzyme-free ethanol sensor based on electrospun nickel nanoparticle-loaded carbon fiber paste electrode.

PubMed

Liu, Yang; Zhang, Lei; Guo, Qiaohui; Hou, Haoqing; You, Tianyan

2010-03-24

We have developed a novel nickel nanoparticle-loaded carbon fiber paste (NiCFP) electrode for enzyme-free determination of ethanol. An electrospinning technique was used to prepare the NiCF composite with large amounts of spherical nanoparticles firmly embedded in carbon fibers (CF). In application to electroanalysis of ethanol, the NiCFP electrode exhibited high amperometric response and good operational stability. The calibration curve was linear up to 87.5 mM with a detection limit of 0.25 mM, which is superior to that obtained with other transition metal based electrodes. For detection of ethanol present in liquor samples, the values obtained with the NiCFP electrode were in agreement with the ones declared on the label. The attractive analytical performance and simple preparation method make this novel material promising for the development of effective enzyme-free sensors. Copyright 2010 Elsevier B.V. All rights reserved.

Surface-Enhanced Separation of Water from Hydrocarbons: Potential Dewatering Membranes for the Catalytic Fast Pyrolysis of Pine Biomass

DOE PAGES

Engtrakul, Dr. Chaiwat; Hu, Michael Z.; Bischoff, Brian L; ...

2016-01-01

The impact of surface-selective coatings on water permeation through a membrane when exposed to catalytic fast pyrolysis (CFP) vapor products was studied by tailoring the surface properties of the membrane coating from superhydrophilic to superhydrophobic. Our approach utilized high-performance architectured surface-selective (HiPAS) membranes that were inserted after a CFP reactor. At this insertion point, the inner wall surface of a tubular membrane was exposed to a mixture of water and upgraded product vapors, including light gases and deoxygenated hydrocarbons. Under proper membrane operating conditions, a high selectivity for water over 1-ring upgraded biomass pyrolysis hydrocarbons was observed due to amore » surface-enhanced capillary condensation process. Owing to this surface-enhanced effect, HiPAS membranes have the potential to enable high flux separations suggesting that water can be selectively removed from the CFP product vapors.« less

Spectrum of antibody profiles in tuberculous elephants, cervids, and cattle.

PubMed

Lyashchenko, Konstantin P; Gortázar, Christian; Miller, Michele A; Waters, W Ray

2018-02-01

Using multi-antigen print immunoassay and DPP ® VetTB Assay approved in the United States for testing captive cervids and elephants, we analyzed antibody recognition of MPB83 and CFP10/ESAT-6 antigens in Asian elephants (Elephas maximus) infected with Mycobacterium tuberculosis and in white-tailed deer (Odocoileus virginianus), fallow deer (Dama dama), elk (Cervus elaphus), and cattle (Bos taurus) infected with Mycobacterium bovis. Serum IgG reactivity to MPB83 was found in the vast majority of tuberculous cattle and cervid species among which white-tailed deer and elk also showed significant CFP10/ESAT-6 recognition rates with added serodiagnostic value. In contrast, the infected elephants developed antibody responses mainly to CFP10/ESAT-6 with MPB83 reactivity being relatively low. The findings demonstrate distinct patterns of predominant antigen recognition by different animal hosts in tuberculosis. Copyright © 2017 Elsevier B.V. All rights reserved.

Surface-Enhanced Separation of Water from Hydrocarbons: Potential Dewatering Membranes for the Catalytic Fast Pyrolysis of Pine Biomass

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

Engtrakul, Chaiwat; Hu, Michael Z.; Bischoff, Brian L.

2016-10-20

The impact of surface-selective coatings on water permeation through a membrane when exposed to catalytic fast pyrolysis (CFP) vapor products was studied by tailoring the surface properties of the membrane coating from superhydrophilic to superhydrophobic. Our approach used high-performance architectured surface-selective (HiPAS) membranes that were inserted after a CFP reactor. At this insertion point, the inner wall surface of a tubular membrane was exposed to a mixture of water and upgraded product vapors, including light gases and deoxygenated hydrocarbons. Under proper membrane operating conditions, a high selectivity for water over one-ring upgraded biomass pyrolysis hydrocarbons was observed as a resultmore » of a surface-enhanced capillary condensation process. Owing to this surface-enhanced effect, HiPAS membranes have the potential to enable high flux separations, suggesting that water can be selectively removed from the CFP product vapors.« less

Catalyst Residence Time Distributions in Riser Reactors for Catalytic Fast Pyrolysis. Part 2: Pilot-Scale Simulations and Operational Parameter Study

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

Foust, Thomas D.; Ziegler, Jack L.; Pannala, Sreekanth

2017-02-21

Here, wsing the validated simulation model developed in part one of this study for biomass catalytic fast pyrolysis (CFP), we assess the functional utility of using this validated model to assist in the development of CFP processes in fluidized catalytic cracking (FCC) reactors to a commercially viable state. Specifically, we examine the effects of mass flow rates, boundary conditions (BCs), pyrolysis vapor molecular weight variation, and the impact of the chemical cracking kinetics on the catalyst residence times. The factors that had the largest impact on the catalyst residence time included the feed stock molecular weight and the degree ofmore » chemical cracking as controlled by the catalyst activity. Lastly, because FCC reactors have primarily been developed and utilized for petroleum cracking, we perform a comparison analysis of CFP with petroleum and show the operating regimes are fundamentally different.« less

An automated wide-field time-gated optically sectioning fluorescence lifetime imaging multiwell plate reader for high-content analysis of protein-protein interactions

NASA Astrophysics Data System (ADS)

Alibhai, Dominic; Kumar, Sunil; Kelly, Douglas; Warren, Sean; Alexandrov, Yuriy; Munro, Ian; McGinty, James; Talbot, Clifford; Murray, Edward J.; Stuhmeier, Frank; Neil, Mark A. A.; Dunsby, Chris; French, Paul M. W.

2011-03-01

We describe an optically-sectioned FLIM multiwell plate reader that combines Nipkow microscopy with wide-field time-gated FLIM, and its application to high content analysis of FRET. The system acquires sectioned FLIM images in <10 s/well, requiring only ~11 minutes to read a 96 well plate of live cells expressing fluorescent protein. It has been applied to study the formation of immature HIV virus like particles (VLPs) in live cells by monitoring Gag-Gag protein interactions using FLIM FRET of HIV-1 Gag transfected with CFP or YFP. VLP formation results in FRET between closely packed Gag proteins, as confirmed by our FLIM analysis that includes automatic image segmentation.

Differences in the Volume of Pharmaceutical Advertisements between Print General Medical Journals

PubMed Central

Gettings, Jennifer; O'Neill, Braden; Chokshi, Dave A.; Colbert, James A.; Gill, Peter; Lebovic, Gerald; Lexchin, Joel; Persaud, Navindra

2014-01-01

Background Pharmaceutical advertisements have been argued to provide revenue that medical journals require but they are intended to alter prescribing behaviour and they are known to include low quality information. We determined whether a difference exists in the current level of pharmaceutical advertising in print general medical journals, and we estimated the revenue generated from print pharmaceutical advertising. Methods Six print general medical journals in Canada, the United States, and the United Kingdom were sampled between 2007 and 2012. The number of advertisements and other journal content in selected issues of the Canadian Medical Association Journal (CMAJ), Canadian Family Physician (CFP), Journal of the American Medical Association (JAMA), New England Journal of Medicine (NEJM), British Medical Journal (BMJ), and Lancet were determined. Revenue gained from pharmaceutical advertising was estimated using each journal's 2013 advertising price list. Findings The two Canadian journals sampled (CMAJ, CFP) contained five times more advertisements than the two American journals (JAMA, NEJM), and two British journals (BMJ, Lancet) (p<0.0001). The estimated annual revenue from pharmaceutical advertisements ranged from £0.025 million (for Lancet) to £3.8 million (for JAMA). The cost savings due to revenue from pharmaceutical advertising to each individual subscriber ranged from £0.02 (for Lancet) to £3.56 (for CFP) per issue. Conclusion The volume of pharmaceutical advertisements differs between general medical journals, with the two Canadian journals sampled containing the most advertisements. International and temporal variations suggest that there is an opportunity for all general medical journals to reduce the number of pharmaceutical advertisements, explore other sources of revenue, and increase transparency regarding sources of revenue. PMID:24416286

Seasonal distribution of Gambierdiscus spp. in Wakasa Bay, the Sea of Japan, and antagonistic relationships with epiphytic pennate diatoms.

PubMed

Nakada, Mitsutaka; Hatayama, Yuki; Ishikawa, Akira; Ajisaka, Tetsuro; Sawayama, Shigeki; Imai, Ichiro

2018-06-01

The occurrence of the ciguatera fish poisoning (CFP) causative Gambierdiscus spp. was confirmed in the Sea of Japan for the first time in 2009. This paper reports seasonal distribution of Gambierdiscus spp. and epiphytic diatoms in the Sea of Japan. Monitoring results suggested an antagonistic interaction in abundances between epiphytic diatoms and the dinoflagellate Gambierdiscus spp. Allelopathic effects of diatoms were considered to be involved in the competitive phenomenon. Therefore it is hypothesized that cell densities of epiphytic pennate diatoms on macroalgae are a novel determinant affecting the abundance of Gambierdiscus spp. other than sea water temperature, salinity and nutrients. Monitorings of the abundance of epiphytic diatoms would lead us to predict the occurrences of Gambierdiscus spp. blooms in the CFP area, and thereby the CFP risk assessments would be developed. Phylogenetic analyses indicated that Gambierdiscus spp. in the Sea of Japan belonged to Gambierdiscus sp. type 2 which was reported to be non-toxic. Nevertheless, based on morphological characteristics, at least two types of Gambierdiscus spp. were found in the Sea of Japan. It is needed to test the toxicity of the both types of Gambierdiscus recognized in the present study for evaluation of the probability of CFP outbreak risks in the Sea of Japan in the future. Copyright © 2018 Elsevier B.V. All rights reserved.

Upgraded bio-oil production via catalytic fast co-pyrolysis of waste cooking oil and tea residual.

PubMed

Wang, Jia; Zhong, Zhaoping; Zhang, Bo; Ding, Kuan; Xue, Zeyu; Deng, Aidong; Ruan, Roger

2017-02-01

Catalytic fast co-pyrolysis (co-CFP) offers a concise and effective process to achieve an upgraded bio-oil production. In this paper, co-CFP experiments of waste cooking oil (WCO) and tea residual (TR) with HZSM-5 zeolites were carried out. The influences of pyrolysis reaction temperature and H/C ratio on pyrolytic products distribution and selectivities of aromatics were performed. Furthermore, the prevailing synergetic effect of target products during co-CFP process was investigated. Experimental results indicated that H/C ratio played a pivotal role in carbon yields of aromatics and olefins, and with H/C ratio increasing, the synergetic coefficient tended to increase, thus led to a dramatic growth of aromatics and olefins yields. Besides, the pyrolysis temperature made a significant contribution to carbon yields, and the yields of aromatics and olefins increased at first and then decreased at the researched temperature region. Note that 600°C was an optimum temperature as the maximum yields of aromatics and olefins could be achieved. Concerning the transportation fuel dependence and security on fossil fuels, co-CFP of WCO and TR provides a novel way to improve the quality and quantity of pyrolysis bio-oil, and thus contributes bioenergy accepted as a cost-competitive and promising alternative energy. Copyright © 2016 Elsevier Ltd. All rights reserved.

Pacific Ciguatoxin Induces Excitotoxicity and Neurodegeneration in the Motor Cortex Via Caspase 3 Activation: Implication for Irreversible Motor Deficit.

PubMed

Asthana, Pallavi; Zhang, Ni; Kumar, Gajendra; Chine, Virendra Bhagawan; Singh, Kunal Kumar; Mak, Yim Ling; Chan, Leo Lai; Lam, Paul Kwan Sing; Ma, Chi Him Eddie

2018-01-18

Consumption of fish containing ciguatera toxins or ciguatoxins (CTXs) causes ciguatera fish poisoning (CFP). In some patients, CFP recurrence occurs even years after exposure related to CTXs accumulation. Pacific CTX-1 (P-CTX-1) is one of the most potent natural substances known that causes predominantly neurological symptoms in patients; however, the underlying pathogenies of CFP remain unknown. Using clinically relevant neurobehavioral tests and electromyography (EMG) to assess effects of P-CTX-1 during the 4 months after exposure, recurrent motor strength deficit occurred in mice exposed to P-CTX-1. We detected irreversible motor strength deficits accompanied by reduced EMG activity, demyelination, and slowing of motor nerve conduction, whereas control unexposed mice fully recovered in 1 month after peripheral nerve injury. Finally, to uncover the mechanism underlying CFP, we detected reduction of spontaneous firing rate of motor cortical neurons even 6 months after exposure and increased number of glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes. Increased numbers of motor cortical neuron apoptosis were detected by dUTP-digoxigenin nick end labeling assay along with activation of caspase 3. Taken together, our study demonstrates that persistence of P-CTX-1 in the nervous system induces irreversible motor deficit that correlates well with excitotoxicity and neurodegeneration detected in the motor cortical neurons.

Deviation of Chinese Adults' Diet from the Chinese Food Pagoda 2016 and Its Association with Adiposity.

PubMed

Tian, Xu; Huang, Yingying; Wang, Hui

2017-09-08

Changing diet in China contributes to a raising prevalence of overweight and obesity. This study aimed to evaluate the dietary status of Chinese adults (20-59 years old) using the China Food Pagoda (CFP) proposed in the Chinese Dietary Guidelines 2016 (CDG), and investigate the association between adiposity and deviation of real diet from CFP using an ordered logistic regression. Results showed that the consumption of fruits, eggs, meat, and poultry increased significantly during 2004-2011, while the consumption of cereal, potatoes, and beans dropped down significantly during the same period (all p < 0.05). Meanwhile, great disparity was detected between real consumption and recommended intake in CFP. In particular, a deficient intake was found for milk and milk products, eggs, and fruit, while over-consumption was observed for cereal, potatoes and beans, meat and poultry, legumes and nuts, oil, and salt. In addition, over-consumption of cereal, legumes and nuts, and salt, as well as under-consumption of vegetables, and meat and poultry, were associated with a higher risk of having high body mass index (BMI), while lower consumption of cereal, potatoes and beans, eggs, and higher consumption of vegetables contributed to low hazard of overweight/obesity (all p < 0.05). The huge disparity between real consumption and the CFP calls for specific health education campaigns.

Study of an outbreak of ciguatera fish poisoning in Hong Kong.

PubMed

Wong, Chun-Kwan; Hung, Patricia; Lee, Kellie L H; Kam, Kai-Man

2005-10-01

Ciguatera fish poisoning (CFP) has been a significant and increasing public health problem in Hong Kong since 1980s. With growing demand for imported live coral fishes, the number of people who suffered from this disease has also been increasing. An outbreak of CFP in 2004 was the second most prominent in record as compared with the most significant one that occurred in 1998. In 2004, out of a total of 823 reported food poisoning outbreaks involving 3159 persons, 65 incidents (7.9%) affecting 247 people (7.8%) were attributed to CFP. Validated mouse bioassay analysis of surveillance samples revealed that seven samples (13%) were confirmed to be contaminated with ciguatoxins (CTXs). Typical symptoms of CTXs were found in mice injected with 20mg of fish extracts. The causative fishes included Cheilinus undulatus, Epinephelus coioides, Plectropomus areolatus, and Plectropomus leopardus. Most of these CTX-positive samples analyzed had only trace amounts of CTXs in their extract, except a C. undulatus sample which contained a mice lethal dose (2.5MU/20mg ether extract). This fish species was also the major origin of coral fish that caused clusters of CFP in the last quarter of 2004. Cigua-Check analysis of 20 flesh grains from seven CTX-positive fishes, previously confirmed as CTX-positive samples by mouse bioassay, showed that 50% of flesh grains were CTX contaminated.

Effect of torrefaction temperature on lignin macromolecule and product distribution from HZSM-5 catalytic pyrolysis

DOE PAGES

Mahadevan, Ravishankar; Adhikari, Sushil; Shakya, Rajdeep; ...

2016-10-27

Torrefaction is a low-temperature process considered as an effective pretreatment technique to improve the grindability of biomass as well as enhance the production of aromatic hydrocarbons from Catalytic Fast Pyrolysis (CFP). For this paper, this study was performed to understand the effect of torrefaction temperature on structural changes in the lignin macromolecule and its subsequent influence on in-situ CFP process. Lignin extracted from southern pine and switchgrass (via organosolv treatment) was torrefied at four different temperatures (150, 175, 200 and 225 °C) in a tubular reactor. Between the two biomass types studied, lignin from pine appeared to have greater thermalmore » stability during torrefaction when compared with switchgrass lignin. The structural changes in lignin as a result of torrefaction were followed by using FTIR spectroscopy, solid state CP/MAS 13C NMR, 31P NMR spectroscopy and it was found that higher torrefaction temperature (200 and 225 °C) caused polycondensation and de-methoxylation of the aromatic units of lignin. Gel permeation chromatography analysis revealed that polycondensation during torrefaction resulted in an increase in the molecular weight and polydispersity of lignin. The torrefied lignin was subsequently used in CFP experiments using H +ZSM-5 catalyst in a micro-reactor (Py-GC/MS) to understand the effect of torrefaction on the product distribution from pyrolysis. It was observed that although the selectivity of benzene-toluene-xylene compounds from CFP of pine improved from 58.3% (torrefaction temp at 150 °C) to 69.0% (torrefaction temp at 225 °C), the severity of torrefaction resulted in a loss of overall aromatic hydrocarbon yield from 11.6% to 4.9% under same conditions. Torrefaction at higher temperatures also increased the yield of carbonaceous residues from 63.9% to 72.8%. Finally, overall, torrefying lignin caused structural transformations in both type of lignins (switchgrass and pine), which is ultimately detrimental to achieving a higher aromatic hydrocarbon yield from CFP.« less

Prevalence and proposal for cost-effective management of the ciguatera risk in the Noumea fish market, New Caledonia (South Pacific).

PubMed

Clua, Eric; Brena, Pierpaolo F; Lecasble, Côme; Ghnassia, Reine; Chauvet, Claude

2011-11-01

Ciguatera fish poisoning (CFP) is a common intoxication associated with the consumption of reef fish, which constitutes a critical issue for public health in many countries. The complexity of its epidemiology is responsible for the poor management of the risk in tropical fish markets. We used the example of the Noumea fish market in New Caledonia to develop a cost-effective methodology of assessing the CFP risk. We first used published reports and the knowledge of local experts to define a list of potentially poisonous local species, ranked by their ciguatoxic potential. Based on two 1-month surveys in the market, conducted in winters 2008 and 2009, we then calculated the consolidated ratio of biomass of potentially poisonous species vs. total biomass of fish sold on the market. The prevalence of high CFP-risk species in the market was 16.1% and 18.9% in 2008 and 2009, respectively. The most common high CFP risk species were groupers (serranids), king mackerels (scombrids), snappers (lutjanids), barracudas (sphyaraenids), emperors (lethrinids) and wrasses (labrids). The size (age) of the fish also plays a critical role in the potential ciguatoxic risk. According to proposals of average size thresholds provided by experts for high-risk species, we were also able to assess the additional risk induced by the sale of some large fish on the market. The data collected both from experts and from the market allowed us to develop a cost-effective proposal for improving the management of the CFP risk in this market. However, the successful implementation of any regulation aiming to ban some specific species and sizes from the market, with an acceptable economical impact, will require the improvement of the expertise in fish identification by public health officers and, ideally, the commitment of retailers. Copyright © 2011 Elsevier Ltd. All rights reserved.

Recombinant ESAT-6-CFP10 Fusion Protein Induction of Th1/Th2 Cytokines and FoxP3 Expressing Treg Cells in Pulmonary TB

PubMed Central

Jackson-Sillah, Dolly; Cliff, Jacqueline M.; Mensah, Gloria Ivy; Dickson, Emmanuel; Sowah, Sandra; Tetteh, John K A.; Addo, Kwasi K.; Ottenhoff, Tom H. M.; Bothamley, Graham; Dockrell, Hazel M.

2013-01-01

Background Early secretory antigenic target 6 (ESAT-6) and culture filtrate protein 10 (CFP-10) are Mycobacterium tuberculosis (Mtb)–specific antigens that are secreted by actively metabolising bacteria and contribute to the virulence of the bacteria. Their ability to induce Treg and Th2 responses, particularly during the first two weeks of treatment, has not been comprehensively examined to date. The purpose of this work was to characterise Th1, Th2 and Treg responses to rESAT-6-CFP10 fusion protein in TB patients before and during the intensive phase of treatment and in healthy M.bovis BCG vaccinated donors. Methods Forty-six newly diagnosed, HIV-negative, smear-positive pulmonary TB patients and 20 healthy donors were recruited in the UK and Ghana. Their peripheral blood mononuclear cells (PBMC) were used in ex vivo ELISPOT and in vitro cultures to identify immunological parameters of interest. Results The study confirmed that protective immune responses to rESAT-6-CFP10 are impaired in active TB but improved during treatment: circulating antigen-specific IL-4-producing T-cells were increased in untreated TB but declined by two weeks of treatment while the circulating antigen-specific IFN-γ producing T cells which showed a transient rise at one week of treatment, persisted at baseline levels at two months of treatment. In vitro T cell proliferation and IFN-γ production were reduced, while IL-4 and CD4+FoxP3+CD25hi cell expression were increased in response to rESAT-6-CFP10 fusion protein in untreated TB. These responses were reversed during early treatment of TB. Conclusions These observations support further investigations into the possible utility of these parameters as markers of active disease and favourable treatment outcomes. PMID:23826366

Effect of torrefaction temperature on lignin macromolecule and product distribution from HZSM-5 catalytic pyrolysis

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

Mahadevan, Ravishankar; Adhikari, Sushil; Shakya, Rajdeep

Torrefaction is a low-temperature process considered as an effective pretreatment technique to improve the grindability of biomass as well as enhance the production of aromatic hydrocarbons from Catalytic Fast Pyrolysis (CFP). For this paper, this study was performed to understand the effect of torrefaction temperature on structural changes in the lignin macromolecule and its subsequent influence on in-situ CFP process. Lignin extracted from southern pine and switchgrass (via organosolv treatment) was torrefied at four different temperatures (150, 175, 200 and 225 °C) in a tubular reactor. Between the two biomass types studied, lignin from pine appeared to have greater thermalmore » stability during torrefaction when compared with switchgrass lignin. The structural changes in lignin as a result of torrefaction were followed by using FTIR spectroscopy, solid state CP/MAS 13C NMR, 31P NMR spectroscopy and it was found that higher torrefaction temperature (200 and 225 °C) caused polycondensation and de-methoxylation of the aromatic units of lignin. Gel permeation chromatography analysis revealed that polycondensation during torrefaction resulted in an increase in the molecular weight and polydispersity of lignin. The torrefied lignin was subsequently used in CFP experiments using H +ZSM-5 catalyst in a micro-reactor (Py-GC/MS) to understand the effect of torrefaction on the product distribution from pyrolysis. It was observed that although the selectivity of benzene-toluene-xylene compounds from CFP of pine improved from 58.3% (torrefaction temp at 150 °C) to 69.0% (torrefaction temp at 225 °C), the severity of torrefaction resulted in a loss of overall aromatic hydrocarbon yield from 11.6% to 4.9% under same conditions. Torrefaction at higher temperatures also increased the yield of carbonaceous residues from 63.9% to 72.8%. Finally, overall, torrefying lignin caused structural transformations in both type of lignins (switchgrass and pine), which is ultimately detrimental to achieving a higher aromatic hydrocarbon yield from CFP.« less

Vibrio vulnificus quorum-sensing molecule cyclo(Phe-Pro) inhibits RIG-I-mediated antiviral innate immunity.

PubMed

Lee, Wooseong; Lee, Seung-Hoon; Kim, Minwoo; Moon, Jae-Su; Kim, Geon-Woo; Jung, Hae-Gwang; Kim, In Hwang; Oh, Ji Eun; Jung, Hi Eun; Lee, Heung Kyu; Ku, Keun Bon; Ahn, Dae-Gyun; Kim, Seong-Jun; Kim, Kun-Soo; Oh, Jong-Won

2018-04-23

The recognition of pathogen-derived ligands by pattern recognition receptors activates the innate immune response, but the potential interaction of quorum-sensing (QS) signaling molecules with host anti-viral defenses remains largely unknown. Here we show that the Vibrio vulnificus QS molecule cyclo(Phe-Pro) (cFP) inhibits interferon (IFN)-β production by interfering with retinoic-acid-inducible gene-I (RIG-I) activation. Binding of cFP to the RIG-I 2CARD domain induces a conformational change in RIG-I, preventing the TRIM25-mediated ubiquitination to abrogate IFN production. cFP enhances susceptibility to hepatitis C virus (HCV), as well as Sendai and influenza viruses, each known to be sensed by RIG-I but did not affect the melanoma-differentiation-associated gene 5 (MDA5)-recognition of norovirus. Our results reveal an inter-kingdom network between bacteria, viruses and host that dysregulates host innate responses via a microbial quorum-sensing molecule modulating the response to viral infection.

[Toxin profiles in fish implicated in ciguatera fish poisoning in Amami and Kakeroma Islands, Kagoshima Prefecture, Japan].

PubMed

Yogi, Kentaro; Oshiro, Naomasa; Matsuda, Seiko; Sakugawa, Satsuki; Matsuo, Toshiaki; Yasumoto, Takeshi

2013-01-01

Ciguatoxins (CTXs) responsible for ciguatera fish poisoning (CFP) in Amami Islands, Kagoshima, Japan in 2008 were determined by LC-MS/MS analysis. Ciguatoxin-1B (CTX1B), 54-deoxyCTX1B, and 52-epi-54-deoxyCTX1B were detected in Variola louti and Lutjanus monostigma. The toxin profile distinctly differed from that of a CFP-related fish from Miyazaki, which mainly contained ciguatoxin-3C type toxins. Toxin profiles were species-specific, as observed in fish from Okinawa. The LC-MS/MS and mouse bioassay (MBA) methods produced comparable data, though 54-deoxyCTX1B was not taken into consideration owing to the lack of toxicity data. To improve assessment, toxicity data for this compound are needed. A reef fish caught on the same occasion and judged nontoxic by MBA (<0.025 MU/g) was found to contain low levels of CTX, indicating a potential risk for CFP.

Chromophore Isomer Stabilization Is Critical to the Efficient Fluorescence of Cyan Fluorescent Proteins.

PubMed

Gotthard, Guillaume; von Stetten, David; Clavel, Damien; Noirclerc-Savoye, Marjolaine; Royant, Antoine

2017-12-12

ECFP, the first usable cyan fluorescent protein (CFP), was obtained by adapting the tyrosine-based chromophore environment in green fluorescent protein to that of a tryptophan-based one. This first-generation CFP was superseded by the popular Cerulean, CyPet, and SCFP3A that were engineered by rational and random mutagenesis, yet the latter CFPs still exhibit suboptimal properties of pH sensitivity and reversible photobleaching behavior. These flaws were serendipitously corrected in the third-generation CFP mTurquoise and its successors without an obvious rationale. We show here that the evolution process had unexpectedly remodeled the chromophore environment in second-generation CFPs so they would accommodate a different isomer, whose formation is favored by acidic pH or light irradiation and which emits fluorescence much less efficiently. Our results illustrate how fluorescent protein engineering based solely on fluorescence efficiency optimization may affect other photophysical or physicochemical parameters and provide novel insights into the rational evolution of fluorescent proteins with a tryptophan-based chromophore.

Noninvasive Evaluation of Heavy Metal Uptake and Storage in Micoralgae Using a Fluorescence Resonance Energy Transfer-Based Heavy Metal Biosensor1[C][W][OPEN

PubMed Central

Rajamani, Sathish; Torres, Moacir; Falcao, Vanessa; Ewalt Gray, Jaime; Coury, Daniel A.; Colepicolo, Pio; Sayre, Richard

2014-01-01

We have developed a fluorescence resonance energy transfer (FRET)-based heavy metal biosensor for the quantification of bioavailable free heavy metals in the cytoplasm of the microalga Chlamydomonas reinhardtii. The biosensor is composed of an end-to-end fusion of cyan fluorescent protein (CFP), chicken metallothionein II (MT-II), and yellow fluorescent protein (YFP). In vitro measurements of YFP/CFP fluorescence emission ratios indicated that the addition of metals to the purified biosensor enhanced FRET between CFP and YFP, consistent with heavy metal-induced folding of MT-II. A maximum YFP/CFP FRET ratio of 2.8 was observed in the presence of saturating concentrations of heavy metals. The sensitivity of the biosensor was greatest for Hg2+ followed by Cd2+ ≈ Pb2+ > Zn2+ > Cu2+. The heavy metal biosensor was unresponsive to metals that do not bind to MT-II (Na+ and Mg2+). When expressed in C. reinhardtii, we observed a differential metal-dependent response to saturating external concentrations (1.6 mm) of heavy metals (Pb2+ > Cd2+) that was unlike that observed for the isolated biosensor (in vitro). Significantly, analysis of metal uptake kinetics indicated that equilibration of the cytoplasm with externally applied heavy metals occurred within seconds. Our results also indicated that algae have substantial buffering capacity for free heavy metals in their cytosol, even at high external metal concentrations. PMID:24368336

Noninvasive evaluation of heavy metal uptake and storage in micoralgae using a fluorescence resonance energy transfer-based heavy metal biosensor.

PubMed

Rajamani, Sathish; Torres, Moacir; Falcao, Vanessa; Ewalt Gray, Jaime; Coury, Daniel A; Colepicolo, Pio; Sayre, Richard

2014-02-01

We have developed a fluorescence resonance energy transfer (FRET)-based heavy metal biosensor for the quantification of bioavailable free heavy metals in the cytoplasm of the microalga Chlamydomonas reinhardtii. The biosensor is composed of an end-to-end fusion of cyan fluorescent protein (CFP), chicken metallothionein II (MT-II), and yellow fluorescent protein (YFP). In vitro measurements of YFP/CFP fluorescence emission ratios indicated that the addition of metals to the purified biosensor enhanced FRET between CFP and YFP, consistent with heavy metal-induced folding of MT-II. A maximum YFP/CFP FRET ratio of 2.8 was observed in the presence of saturating concentrations of heavy metals. The sensitivity of the biosensor was greatest for Hg2+ followed by Cd2+≈Pb2+>Zn2+>Cu2+. The heavy metal biosensor was unresponsive to metals that do not bind to MT-II (Na+ and Mg2+). When expressed in C. reinhardtii, we observed a differential metal-dependent response to saturating external concentrations (1.6 mm) of heavy metals (Pb2+>Cd2+) that was unlike that observed for the isolated biosensor (in vitro). Significantly, analysis of metal uptake kinetics indicated that equilibration of the cytoplasm with externally applied heavy metals occurred within seconds. Our results also indicated that algae have substantial buffering capacity for free heavy metals in their cytosol, even at high external metal concentrations.

Differences in immune cell function between tuberculosis positive and negative Asian elephants.

PubMed

Landolfi, Jennifer A; Miller, Michele; Maddox, Carol; Zuckermann, Federico; Langan, Jennifer N; Terio, Karen A

2014-07-01

Tuberculosis is an important health concern for Asian elephant (Elephas maximus) populations worldwide, however, mechanisms underlying susceptibility to Mycobacterium tuberculosis are unknown. Proliferative responses assessed via brominated uridine incorporation and cytokine expression measured by real-time RT-PCR were evaluated in peripheral blood mononuclear cell (PBMC) cultures from 8 tuberculosis negative and 8 positive Asian elephants. Cultures were stimulated with Mycobacterium bovis purified protein derivative (PPD-B), M. tuberculosis culture filtrate protein (CFP)-10, and Mycobacterium avium PPD (PPD-A). Following stimulation with PPD-B, proliferation was higher (α = 0.005) in positive samples; no significant differences were detected following CFP-10 or PPD-A stimulation. Tumor necrosis factor (TNF)-α, interleukin (IL)-12, and interferon (IFN)-γ expression was greater in samples from positive elephants following stimulation with PPD-B (α = 0.025) and CFP-10 (α = 0.025 TNF-α and IL-12; α = 0.005 IFN-γ). Stimulation with PPD-A also produced enhanced IL-12 expression in positive samples (α = 0.025). Findings suggested that differences in immune cell function exist between tuberculosis positive and negative elephants. Proliferative responses and expression of TNF-α, IL-12, and IFN-γ in response to stimulation with PPD-B and CFP-10 differ between tuberculosis positive and negative elephants, suggesting these parameters may be important to tuberculosis immunopathogenesis in this species. Copyright © 2014 Elsevier Ltd. All rights reserved.

A FRET-Based Method for Probing the Conformational Behavior of an Intrinsically Disordered Repeat Domain from Bordetella pertussis Adenylate Cyclase

DTIC Science & Technology

2009-10-22

ELEMENT NUMBER 6. AUTHOR( S ) 5d. PROJECT NUMBER 5e. TASK NUMBER 5f. WORK UNIT NUMBER 7. PERFORMING ORGANIZATION NAME( S ) AND ADDRESS(ES...MONITORING AGENCY NAME( S ) AND ADDRESS(ES) 10. SPONSOR/MONITOR’S ACRONYM( S ) 11. SPONSOR/MONITOR’S REPORT NUMBER( S ) 12. DISTRIBUTION/AVAILABILITY STATEMENT...fusion protein C*-BR( S )-Y* expression vector pET/C*-CyaA1488-1680-Y*, nonfluorescent CFP expression vector pET/CFP*, and the maltose binding protein-RTX

IFNγ Response to Mycobacterium tuberculosis, Risk of Infection and Disease in Household Contacts of Tuberculosis Patients in Colombia

PubMed Central

Marín, Nancy D.; Marín, Diana M.; López, Lucelly; Henao, Hanna M.; Martínez, Teresita; Villa, Liliana; Barrera, Luis F.; Ortiz, Blanca L.; Ramírez, María E.; Montes, Carlos J.; Oquendo, María C.; Arango, Lisandra M.; Riaño, Felipe; Aguirre, Carlos; Bustamante, Alberto; Belisle, John T.; Dobos, Karen; Mejía, Gloria I.; Giraldo, Margarita R.; Brennan, Patrick J.; Robledo, Jaime; Arbeláez, María P.; Rojas, Carlos A.; García, Luis F.

2009-01-01

Objectives Household contacts (HHCs) of pulmonary tuberculosis patients are at high risk of Mycobacterium tuberculosis infection and early disease development. Identification of individuals at risk of tuberculosis disease is a desirable goal for tuberculosis control. Interferon-gamma release assays (IGRAs) using specific M. tuberculosis antigens provide an alternative to tuberculin skin testing (TST) for infection detection. Additionally, the levels of IFNγ produced in response to these antigens may have prognostic value. We estimated the prevalence of M. tuberculosis infection by IGRA and TST in HHCs and their source population (SP), and assessed whether IFNγ levels in HHCs correlate with tuberculosis development. Methods A cohort of 2060 HHCs was followed for 2–3 years after exposure to a tuberculosis case. Besides TST, IFNγ responses to mycobacterial antigens: CFP, CFP-10, HspX and Ag85A were assessed in 7-days whole blood cultures and compared to 766 individuals from the SP in Medellín, Colombia. Isoniazid prophylaxis was not offered to child contacts because Colombian tuberculosis regulations consider it only in children under 5 years, TST positive without BCG vaccination. Results Using TST 65.9% of HHCs and 42.7% subjects from the SP were positive (OR 2.60, p<0.0001). IFNγ response to CFP-10, a biomarker of M. tuberculosis infection, tested positive in 66.3% HHCs and 24.3% from the SP (OR = 6.07, p<0.0001). Tuberculosis incidence rate was 7.0/1000 person years. Children <5 years accounted for 21.6% of incident cases. No significant difference was found between positive and negative IFNγ responders to CFP-10 (HR 1.82 95% CI 0.79–4.20 p = 0.16). However, a significant trend for tuberculosis development amongst high HHC IFNγ producers was observed (trend Log rank p = 0.007). Discussion CFP-10-induced IFNγ production is useful to establish tuberculosis infection prevalence amongst HHC and identify those at highest risk of disease. The high tuberculosis incidence amongst children supports administration of chemoprohylaxis to child contacts regardless of BCG vaccination. PMID:20011589

"One-for-All" Strategy in Fast Energy Storage: Production of Pillared MOF Nanorod-Templated Positive/Negative Electrodes for the Application of High-Performance Hybrid Supercapacitor.

PubMed

Qu, Chong; Liang, Zibin; Jiao, Yang; Zhao, Bote; Zhu, Bingjun; Dang, Dai; Dai, Shuge; Chen, Yu; Zou, Ruqiang; Liu, Meilin

2018-06-01

Currently, metal-organic frameworks (MOFs) are intensively studied as active materials for electrochemical energy storage applications due to their tunable structure and exceptional porosities. Among them, water stable pillared MOFs with dual ligands have been reported to exhibit high supercapacitor (SC) performance. Herein, the "One-for-All" strategy is applied to synthesize both positive and negative electrodes of a hybrid SC (HSC) from a single pillared MOF. Specifically, Ni-DMOF-TM ([Ni(TMBDC)(DABCO) 0.5 ], TMBDC: 2,3,5,6-tetramethyl-1,4-benzenedicarboxylic acid, DABCO: 1,4-diazabicyclo[2.2.2]-octane) nanorods are directly grown on carbon fiber paper (CFP) (denoted as CFP@TM-nanorods) with the help of triethylamine and function as the positive electrode of HSC under alkaline electrolyte. Meanwhile, calcinated N-doped hierarchical porous carbon nanorods (CFP@TM-NPCs) are produced and utilized as the negative counter-electrode from a one-step heat treatment of CFP@TM-nanorods. After assembling these two electrodes together to make a hybrid device, the TM-nanorods//TM-NPCs exhibit a wide voltage window of 1.5 V with a high sloping discharge plateau between 1-1.2 V, indicating its great potential for practical applications. This as-described "One-for-All" strategy is widely applicable and highly reproducible in producing MOF-based electrode materials for HSC applications, which shortens the gap between experimental synthesis and practical application of MOFs in fast energy storage. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

In-situ Electrodeposition of Highly Active Silver Catalyst on Carbon Fiber Papers as Binder Free Cathodes for Aluminum-air Battery.

PubMed

Hong, Qingshui; Lu, Huimin

2017-06-13

Carbon fiber papers supported Ag catalysts (Ag/CFP) with different coverage of electro-active site are prepared by electrochemical deposition and used as binder free cathodes in primary aluminum-air (Al-air) battery. Scanning Electron Microscopy and X-ray Diffraction studies are carried out to characterize the as-prepared Ag/CFP air cathodes. Oxygen reduction reaction (ORR) activities on these air cathodes in alkaline solutions are systematic studied. A newly designed aluminum-air cell is used to further determine the cathodes performance under real operation condition and during the test, the Ag/CFP electrodes show outstanding catalytic activity for ORR in concentrated alkaline electrolyte, and no obvious activity degradation is observed after long-time discharge. The electrochemical test results display the dependence of coverage of the electro-active Ag on the catalytic performance of the air cathodes. The resulting primary Al-air battery made from the best-performing cathode shows an impressive discharge peak power density, outperforming that of using commercial nano-manganese catalyst air electrodes.

Evaluating patient acceptability of a culturally focused psychiatric consultation intervention for Latino Americans with depression.

PubMed

Trinh, Nhi-Ha T; Hagan, Patrick N; Flaherty, Katherine; Traeger, Lara N; Inamori, Aya; Brill, Charlotte D; Hails, Katherine; Chang, Trina E; Bedoya, C Andres; Fava, Maurizio; Yeung, Albert

2014-12-01

Significant disparities exist in both access to and quality of mental health care for Latino Americans with depression compared to Caucasians, resulting in a greater burden of disability in this underserved population. Our aim is to evaluate participant acceptability of a Culturally Focused Psychiatric (CFP) consultation program for depressed Latino Americans. Latino American adult primary care patients endorsing depressive symptoms on a screening questionnaire were targeted in their primary care clinic. The intervention addressed participants' depressive symptoms using culturally adapted clinical assessments and toolkits. Acceptability was evaluated using a treatment satisfaction scale and in-depth semi-structured interviews. Overall, 85% of participants responded positively to all questions of the satisfaction scale. In in-depth interviews, the vast majority of participants reported the program met expectations, all stated providers were culturally sensitive, and most stated recommendations were culturally sensitive. The CFP program was found to be acceptable to a group of depressed Latino American primary care patients. Further research is needed to evaluate if the CFP intervention can improve depressive symptoms and outcomes.

The SET1 Complex Selects Actively Transcribed Target Genes via Multivalent Interaction with CpG Island Chromatin.

PubMed

Brown, David A; Di Cerbo, Vincenzo; Feldmann, Angelika; Ahn, Jaewoo; Ito, Shinsuke; Blackledge, Neil P; Nakayama, Manabu; McClellan, Michael; Dimitrova, Emilia; Turberfield, Anne H; Long, Hannah K; King, Hamish W; Kriaucionis, Skirmantas; Schermelleh, Lothar; Kutateladze, Tatiana G; Koseki, Haruhiko; Klose, Robert J

2017-09-05

Chromatin modifications and the promoter-associated epigenome are important for the regulation of gene expression. However, the mechanisms by which chromatin-modifying complexes are targeted to the appropriate gene promoters in vertebrates and how they influence gene expression have remained poorly defined. Here, using a combination of live-cell imaging and functional genomics, we discover that the vertebrate SET1 complex is targeted to actively transcribed gene promoters through CFP1, which engages in a form of multivalent chromatin reading that involves recognition of non-methylated DNA and histone H3 lysine 4 trimethylation (H3K4me3). CFP1 defines SET1 complex occupancy on chromatin, and its multivalent interactions are required for the SET1 complex to place H3K4me3. In the absence of CFP1, gene expression is perturbed, suggesting that normal targeting and function of the SET1 complex are central to creating an appropriately functioning vertebrate promoter-associated epigenome. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Dimerization between aequorea fluorescent proteins does not affect interaction between tagged estrogen receptors in living cells

PubMed Central

Kofoed, Eric M.; Guerbadot, Martin; Schaufele, Fred

2008-01-01

Förster resonance energy transfer (FRET) detection of protein interaction in living cells is commonly measured following the expression of interacting proteins genetically fused to the cyan (CFP) and yellow (YFP) derivatives of the Aequorea victoria fluorescent protein (FP). These FPs can dimerize at mM concentrations, which may introduce artifacts into the measurement of interaction between proteins that are fused with the FPs. Here, FRET analysis of the interaction between estrogen receptors (alpha isoform, ERα) labeled with “wild-type” CFP and YFP is compared with that of ERα labeled with “monomeric” A206K mutants of CFP and YFP. The intracellular equilibrium dissociation constant for the hormone-induced ERα-ERα interaction is similar for ERα labeled with wild-type or monomeric FPs. However, the measurement of energy transfer measured for ERα-ERα interaction in each cell is less consistent with the monomeric FPs. Thus, dimerization of the FPs does not affect the kinetics of ERα-ERα interaction but, when brought close together via ERα-ERα interaction, FP dimerization modestly improves FRET measurement. PMID:18601531

Implementation and assessment of a fast-track programme to improve communication between primary and specialized care in patients with suspected cancer: how to shorten time between initial symptoms of cancer, diagnosis and initiation of treatment.

PubMed

Martínez, M T; González, I; Tarazona, N; Roselló, S; Saiz, R; Sanmartín, A; Martínez-Agulló, Á; Caballero, A; Mas, P; Franco, J; Martínez-Jabaloyas, J; García-Callejo, J; Martín, V; Navarro, J; Teruel, A; Lluch, A; Chirivella, I

2015-02-01

This study aims to asses a cancer fast-track programme (CFP) to shorten the time since a patient with suspicion of cancer is referred by the primary care (PC) physician to the specialized medical team. Guidelines for main suspected tumours were designed to help PC physicians to detect and rapidly refer cases to the CFP oncology coordinator, who sent them to the appropriate department to accelerate diagnosis, staging and therapy. All patients analysed in this report were referred from June 2009 to July 2012. A total of 897 suspected cancer cases were submitted and finally 705 were studied. In 205 (29 %) a cancer diagnosis was confirmed within 23 days (median). Therapy was initiated within 46 days after referral (median). Early diagnoses with a potential curative approach were made in 166 (82 %). This CFP decreased the waiting time for cancer diagnosis, by improving communication between PC physician and specialized care teams. Most patients included in this program could get therapy with curative intent.

Hydrogen peroxide sensor based on carbon nanowalls grown by plasma-enhanced chemical vapor deposition

NASA Astrophysics Data System (ADS)

Tomatsu, Masakazu; Hiramatsu, Mineo; Foord, John S.; Kondo, Hiroki; Ishikawa, Kenji; Sekine, Makoto; Takeda, Keigo; Hori, Masaru

2017-06-01

Fabrication of an electrochemical sensor for hydrogen peroxide (H2O2) detection was demonstrated. H2O2 is a major messenger molecule in various redox-dependent cellular signaling transductions. Therefore, sensitive detection of H2O2 is greatly important in health inspection and environmental protection. Carbon nanowalls (CNWs) are composed of few-layer graphenes standing almost vertically on a substrate forming a three-dimensional structure. In this work, CNWs were used as a platform for H2O2 sensing, which is based on the large surface area of conducting carbon and surface decoration with platinum (Pt) nanoparticles (NPs). CNWs were grown on carbon fiber paper (CFP) by inductively coupled plasma-enhanced chemical vapor deposition to increase the surface area. Then, the CNW surface was decorated with Pt-NPs by the reduction of H2PtCl6. Cyclic voltammetry results indicate that the Pt-decorated CNW/CFP electrode possesses excellent electrocatalytic activity for the reduction of H2O2. Amperometric responses indicate the high-sensitivity detection capability of the Pt-decorated CNW/CFP electrode for H2O2.

Ciguatera fish poisoning on the West Africa Coast: An emerging risk in the Canary Islands (Spain).

PubMed

Boada, Luis D; Zumbado, Manuel; Luzardo, Octavio P; Almeida-González, Maira; Plakas, Steven M; Granade, Hudson R; Abraham, Ann; Jester, Edward L E; Dickey, Robert W

2010-12-01

Ciguatera fish poisoning (CFP) is endemic in certain tropical and subtropical regions of the world. CFP had not been described on the West Africa Coast until a 2004 outbreak in the Canary Islands. In 2008-2009, two additional outbreaks of ciguatera occurred. Individuals afflicted had consumed lesser amberjack (Seriola rivoliana) captured from nearby waters. Caribbean ciguatoxin-1 (C-CTX-1) was confirmed in fish samples by LC-MS/MS. Ciguatoxic fish in this region may pose a new health risk for the seafood consumer. Copyright © 2010 Elsevier Ltd. All rights reserved.

Three-color confocal Förster (or fluorescence) resonance energy transfer microscopy: Quantitative analysis of protein interactions in the nucleation of actin filaments in live cells.

PubMed

Wallrabe, Horst; Sun, Yuansheng; Fang, Xiaolan; Periasamy, Ammasi; Bloom, George S

2015-06-01

Experiments using live cell 3-color Förster (or fluorescence) resonance energy transfer (FRET) microscopy and corresponding in vitro biochemical reconstitution of the same proteins were conducted to evaluate actin filament nucleation. A novel application of 3-color FRET data is demonstrated, extending the analysis beyond the customary energy-transfer efficiency (E%) calculations. MDCK cells were transfected for coexpression of Teal-N-WASP/Venus-IQGAP1/mRFP1-Rac1, Teal-N-WASP/Venus-IQGAP1/mRFP1-Cdc42, CFP-Rac1/Venus-IQGAP1/mCherry-actin, or CFP-Cdc42/Venus-IQGAP1/mCherry-actin, and with single-label equivalents for spectral bleedthrough correction. Using confirmed E% as an entry point, fluorescence levels and related ratios were correlated at discrete accumulating levels at cell peripheries. Rising ratios of CFP-Rac1:Venus-IQGAP1 were correlated with lower overall actin fluorescence, whereas the CFP-Cdc42:Venus-IQGAP1 ratio correlated with increased actin fluorescence at low ratios, but was neutral at higher ratios. The new FRET analyses also indicated that rising levels of mRFP1-Cdc42 or mRFP1-Rac1, respectively, promoted or suppressed the association of Teal-N-WASP with Venus-IQGAP1. These 3-color FRET assays further support our in vitro results about the role of IQGAP1, Rac1, and Cdc42 in actin nucleation, and the differential impact of Rac1 and Cdc42 on the association of N-WASP with IQGAP1. In addition, this study emphasizes the power of 3-color FRET as a systems biology strategy for simultaneous evaluation of multiple interacting proteins in individual live cells. © 2015 International Society for Advancement of Cytometry.

Conditional ablation of osteoblasts in medaka.

PubMed

Willems, Bernd; Büttner, Anita; Huysseune, Ann; Renn, Joerg; Witten, P Eckhard; Winkler, Christoph

2012-04-15

Different from tetrapods, teleost vertebral centra form without prior establishment of a cartilaginous scaffold, in two steps: First, mineralization of the notochord sheath establishes the vertebral centra. Second, sclerotome derived mesenchymal cells migrate around the notochord sheath. These cells differentiate into osteoblasts and deposit bone onto the mineralized notochord sheath in a process of intramembranous bone formation. In contrast, most skeletal elements of the cranial skeleton arise by chondral bone formation, with remarkably similar mechanisms in fish and tetrapods. To further investigate the role of osteoblasts during formation of the cranial and axial skeleton, we generated a transgenic osx:CFP-NTR medaka line which enables conditional ablation of osterix expressing osteoblasts. By expressing a bacterial nitroreductase (NTR) fused to Cyan Fluorescent Protein (CFP) under control of the osterix promoter these cells become sensitive towards Metronidazole (Mtz). Mtz treatment of stable osx:CFP-NTR transgenic medaka for several consecutive days led to significant loss of osteoblasts by apoptosis. Live staining of mineralized bone matrix revealed reduced ossification in head skeletal elements such as cleithrum and operculum, as well as in the vertebral arches. Interestingly in Mtz treated larvae, intervertebral spaces were missing and the notochord sheath was often continuously mineralized resulting in the fusion of centra. We therefore propose a dual role for osx-positive osteoblasts in fish. Besides a role in bone deposition, we suggest an additional border function during mineralization of the chordal centra. After termination of Mtz treatment, osteoblasts gradually reappeared, indicating regenerative properties in this cell lineage. Taken together, the osx:CFP-NTR medaka line represents a valuable tool to study osteoblast function and regeneration at different stages of development in whole vertebrate specimens in vivo. Copyright © 2012 Elsevier Inc. All rights reserved.

Transgenic nude mice ubiquitously expressing fluorescent proteins for color-coded imaging of the tumor microenvironment.

PubMed

Hoffman, Robert M

2014-01-01

We have developed a transgenic green fluorescent protein (GFP) nude mouse with ubiquitous GFP expression. The GFP nude mouse was obtained by crossing nontransgenic nude mice with the transgenic C57/B6 mouse in which the β-actin promoter drives GFP expression in essentially all tissues. In the adult mice, many organs brightly expressed GFP, including the spleen, heart, lungs, spleen, pancreas, esophagus, stomach, and duodenum as well as the circulatory system. The liver expressed GFP at a lesser level. The red fluorescent protein (RFP) transgenic nude mouse was obtained by crossing non-transgenic nude mice with the transgenic C57/B6 mouse in which the beta-actin promoter drives RFP (DsRed2) expression in essentially all tissues. In the RFP nude mouse, the organs all brightly expressed RFP, including the heart, lungs, spleen, pancreas, esophagus, stomach, liver, duodenum, the male and female reproductive systems; brain and spinal cord; and the circulatory system, including the heart, and major arteries and veins. The skinned skeleton highly expressed RFP. The bone marrow and spleen cells were also RFP positive. The cyan fluorescent protein (CFP) nude mouse was developed by crossing nontransgenic nude mice with the transgenic CK/ECFP mouse in which the β-actin promoter drives expression of CFP in almost all tissues. In the CFP nude mice, the pancreas and reproductive organs displayed the strongest fluorescence signals of all internal organs, which vary in intensity. The GFP, RFP, and CFP nude mice when transplanted with cancer cells of another color are powerful models for color-coded imaging of the tumor microenvironment (TME) at the cellular level.

Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein: structural characterization and applications in fluorescence imaging

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

Al, Hui-wang; Henderson, J. Nathan; Remington, S. James

The arsenal of engineered variants of the GFP [green FP (fluorescent protein)] from Aequorea jellyfish provides researchers with a powerful set of tools for use in biochemical and cell biology research. The recent discovery of diverse FPs in Anthozoa coral species has provided protein engineers with an abundance of alternative progenitor FPs from which improved variants that complement or supersede existing Aequorea GFP variants could be derived. Here, we report the engineering of the first monomeric version of the tetrameric CFP (cyan FP) cFP484 from Clavularia coral. Starting from a designed synthetic gene library with mammalian codon preferences, we identifiedmore » dimeric cFP484 variants with fluorescent brightness significantly greater than the wild-type protein. Following incorporation of dimer-breaking mutations and extensive directed evolution with selection for blue-shifted emission, high fluorescent brightness and photostability, we arrived at an optimized variant that we have named mTFP1 [monomeric TFP1 (teal FP 1)]. The new mTFP1 is one of the brightest and most photostable FPs reported to date. In addition, the fluorescence is insensitive to physiologically relevant pH changes and the fluorescence lifetime decay is best fitted as a single exponential. The 1.19 {angstrom} crystal structure (1 {angstrom}=0.1 nm) of mTFP1 confirms the monomeric structure and reveals an unusually distorted chromophore conformation. As we experimentally demonstrate, the high quantum yield of mTFP1 (0.85) makes it particularly suitable as a replacement for ECFP (enhanced CFP) or Cerulean as a FRET (fluorescence resonance energy transfer) donor to either a yellow or orange FP acceptor.« less

Quantitative FRET imaging of leptin receptor oligomerization kinetics in single cells.

PubMed

Biener, Eva; Charlier, Madia; Ramanujan, V Krishnan; Daniel, Nathalie; Eisenberg, Avital; Bjørbaek, Christian; Herman, Brian; Gertler, Arieh; Djiane, Jean

2005-12-01

Leptin, an adipocyte-secreted hormone, signals through activation of its membrane-embedded receptor (LEPR). To study the leptin-induced events occurring in short (LEPRa) and long (LEPRb) LEPRs in the cell membrane, by FRET (fluorescence resonance energy transfer) methodology, the respective receptors, tagged at their C-terminal with CFP (cyan fluorescent protein) or YFP (yellow fluorescent protein), were prepared. The constructs encoding mLEPRa (mouse LEPRa)-YFP and mLEPRa-CFP, mLEPRb-YFP and mLEPRb-CFP were tested for biological activity in transiently transfected CHO cells (Chinese-hamster ovary cells) and HEK-293T cells (human embryonic kidney 293 T cells) for activation of STAT3 (signal transduction and activators of transcription 3)-mediated LUC (luciferase) activity and binding of radiolabelled leptin. All four constructs were biologically active and were as potent as their untagged counterparts. The localization pattern of the fused protein appeared to be confined almost entirely to the cell membrane. The leptin-dependent interaction between various types of receptors in fixed cells were studied by measuring FRET, using fluorescence lifetime imaging microscopy and acceptor photobleaching methods. Both methods yielded similar results, indicating that (1) leptin receptors expressed in the cell membrane exist mostly as preformed LEPRa/LEPRa or LEPRb/LEPRb homo-oligomers but not as LEPRb/LEPRa hetero-oligomers; (2) the appearance of transient leptin-induced FRET in cells transfected with LEPRb/LEPRb reflects both a conformational change that leads to closer interaction in the cytosolic part and a higher FRET signal, as well as de novo homo-oligomerization; (3) in LEPRa/LEPRa, exposure to leptin does not lead to any increase in FRET signalling as the proximity of CFP and YFP fluorophores in space already gives maximal FRET efficiency of the preoligomerized receptors.

Combinatorial usage of fungal polysaccharides from Cordyceps sinensis and Ganoderma atrum ameliorate drug-induced liver injury in mice.

PubMed

Fan, Songtao; Huang, Xiaojun; Wang, Sunan; Li, Chang; Zhang, Zhihong; Xie, Mingyong; Nie, Shaoping

2018-05-15

This study investigated the possible protective effect of combined fungal polysaccharides (CFP), consisting of Cordyceps sinensis polysaccharides (CSP) and Ganoderma atrum polysaccharides (PSG) with well-defined structural characteristics, against cyclophosphamide (CTX)-induced hepatotoxicity in mice. Our results indicated CFP effectively prevented the liver injury by decreasing toxicity markers (aspartate transaminase, alanine aminotransferase and alkaline phosphatase). Further biochemical and molecular analysis indicated CSP particularly inhibited the activation of Toll-like receptor 9 (TLR9) and its related inflammatory signals, including pro-inflammatory cytokines, inducible nitric oxide synthase, and cyclooxygenase-2 to modulate hepatic inflammation response. Relatively, through activation of peroxisome proliferator-activated receptor α (PPARα), PSG increased hepatic glutathione peroxidase and glutathione content depleted by CTX, as well as prevented mitochondria-dependent apoptosis with regulation on Bcl-2 family proteins (Bad, Bax and Bcl-2). In addition, protective effect of CFP was associated with enhanced modulations on cellular oxidant/antioxidant imbalance, mitochondrial apoptotic pathway and pro-inflammatory factors via PPARα upregulation and TLR9 downregulation. Taking together, the combinatorial approach based on CSP and PSG presented a practical option for the management of drug-induced liver injury. Copyright © 2018 Elsevier Ltd. All rights reserved.

Functional Class in Children with Idiopathic Dilated Cardiomyopathy. A pilot Study

PubMed Central

Tavares, Aline Cristina; Bocchi, Edimar Alcides; Guimarães, Guilherme Veiga

2016-01-01

Background Idiopathic dilated cardiomyopathy (IDCM), most common cardiac cause of pediatric deaths, mortality descriptor: a low left ventricular ejection fraction (LVEF) and low functional capacity (FC). FC is never self reported by children. Objective The aims of this study were (i) To evaluate whether functional classifications according to the children, parents and medical staff were associated. (iv) To evaluate whether there was correlation between VO2 max and Weber's classification. Method Prepubertal children with IDCM and HF (by previous IDCM and preserved LVEF) were selected, evaluated and compared. All children were assessed by testing, CPET and functional class classification. Results Chi-square test showed association between a CFm and CFp (1, n = 31) = 20.6; p = 0.002. There was no significant association between CFp and CFc (1, n = 31) = 6.7; p = 0.4. CFm and CFc were not associated as well (1, n = 31) = 1.7; p = 0.8. Weber's classification was associated to CFm (1, n = 19) = 11.8; p = 0.003, to CFp (1, n = 19) = 20.4; p = 0.0001and CFc (1, n = 19) = 6.4; p = 0.04). Conclusion Drawing were helpful for children's self NYHA classification, which were associated to Weber's stratification. PMID:27168472

NASA-OAI Collaborative Aerospace Research and Fellowship Program at NASA Glenn Research Center at Lewis Field

NASA Technical Reports Server (NTRS)

Heyward, Ann O.; Montegani, Francis J.

2003-01-01

During the summer of 2002, a IO-week activity for university faculty entitled the NASA-OAI Collaborative Aerospace Research and Fellowship Program (CFP) was conducted at the NASA Glenn Research Center in collaboration with the Ohio Aerospace Institute (OAI). This is a companion program to the highly successful NASA Faculty Fellowship Program and its predecessor, the NASA- ASEE Summer Faculty Fellowship Program, that operated for 38 years at Glenn. This year s program began officially on June 3, 2002 and continued through August 9, 2002. This report is intended primarily to summarize the research activities comprising the 2002 CFP Program at Glenn. Fifteen research summaries are included.

Effect of Water on Axial Flow Compressors. Part I. Analysis and Predictions.

DTIC Science & Technology

1981-06-01

MAIN 21 REAL NDNUKAM,MMASSMMASSl MAIN 22 REAL MMASSO MAIN 23 COMMON TD(7),IUMIT MAIM 24 COMMON CFL, CFT , CFP, CFD, CFM, CFU, CFA MAIN 25 COMMON JPERFM...MAIN 155 1519 FORMAT(6F5.2) MAIM 156 READ(5,1520) (BET2SS(I)vlIN51) MAIN 157 1520 FORMAT(7F5.2) MAIN 158 CFL=2. 54 MAIM 159 CFT =1.0𔃻.8 MAIM 160 CFP=47...SC( I)*CFL MAIN 183 SRTIP( I )SRTIP( I)*CFL MAIM 184 SAREAS( I )SAREAS( I)*CFA MAIN 185 1570 CONTINUE MAIM 186 T0C=TOG’CFT MAIN 187 TOW=TOW* CFT MAIM

The recombinant fusion protein CFP10-ESAT6-dIFN has protective effect against tuberculosis in guinea pigs.

PubMed

Permyakova, Natalya V; Belavin, Pavel A; Pirozhkova, Dariya S; Ufimtseva, Elena G; Rozov, Sergey M; Mursalimov, Sergey R; Sidorchuk, Yuriy V; Uvarova, Elena A; Zagorskaya, Alla A; Marenkova, Tatiana V; Bannikova, Svetlana V; Demidov, Evgeniy A; Starostin, Konstantin V; Kravchenko, Marionella A; Vakhrusheva, Diana V; Berdnikov, Roman B; Eremeeva, Natalya I; Skornyakov, Sergey N; Peltek, Sergey E; Deineko, Elena V

2018-03-01

Development of effective vaccine candidates against tuberculosis (TB) is currently the most important challenge in the prevention of this disease since the BCG vaccine fails to guarantee a lifelong protection, while any other approved vaccine with better efficiency is still absent. The protective effect of the recombinant fusion protein CFP10-ESAT6-dIFN produced in a prokaryotic expression system (Escherichia coli) has been assessed in a guinea pig model of acute TB. The tested antigen comprises the Mycobacterium tuberculosis (Mtb) proteins ESAT6 and CFP10 as well as modified human γ-interferon (dIFN) for boosting the immune response. Double intradermal immunization of guinea pigs with the tested fusion protein (2 × 0.5 µg) induces a protective effect against subsequent Mtb infection. The immunized guinea pigs do not develop the symptoms of acute TB and their body weight gain was five times more as compared with the non-immunized infected guinea pigs. The animal group immunized with this dose of antigen displays the minimum morphological changes in the internal organs and insignificant inflammatory lesions in the liver tissue, which complies with a decrease in the bacterial load in the spleen and average Mtb counts in macrophages.

Gender differences in body-sway factors of center of foot pressure in a static upright posture and under the influence of alcohol intake.

PubMed

Kitabayashi, Tamotsu; Demura, Shinichi; Noda, Masahiro; Yamada, Takayoshi

2004-07-01

This study aimed to examine gender differences in 4 body-sway factors of the center of foot pressure (CFP) during a static upright posture and the influence of alcohol intake on them. Four body-sway factors were interpreted in previous studies using factor analysis (the principal factor method and oblique solution by promax-rotation) on 220 healthy young males and females as follows; unit time sway, front-back sway, left-right sway and high frequency band power. The CFP measurement for 1 min was carried out twice with 1 min rest. The measurements of blood pressure, heart rate, whole body reaction time, standing on one leg with eyes closed, and CFP were carried out before and after the alcohol intake using 11 healthy young males and females. The measurement device used was an Anima's stabilometer G5500. The data sampling frequency was 20 Hz. Reliability of 4 body-sway factors was very high. Significant gender differences were found in the left-right sway and the high frequency band power factors, but the influence on body-sway is, as a whole, can be disregarded. These four sway factors can determine the influence of alcohol intake as efficient as 32 sway parameters.

Ciguatoxin reduces regenerative capacity of axotomized peripheral neurons and delays functional recovery in pre-exposed mice after peripheral nerve injury.

PubMed

Au, Ngan Pan Bennett; Kumar, Gajendra; Asthana, Pallavi; Tin, Chung; Mak, Yim Ling; Chan, Leo Lai; Lam, Paul Kwan Sing; Ma, Chi Him Eddie

2016-05-27

Ciguatera fish poisoning (CFP) results from consumption of tropical reef fish containing ciguatoxins (CTXs). Pacific (P)-CTX-1 is among the most potent known CTXs and the predominant source of CFP in the endemic region responsible for the majority of neurological symptoms in patients. Chronic and persistent neurological symptoms occur in some CFP patients, which often result in incomplete functional recovery for years. However, the direct effects of exposure to CTXs remain largely unknown. In present study, we exposed mice to CTX purified from ciguatera fish sourced from the Pacific region. P-CTX-1 was detected in peripheral nerves within hours and persisted for two months after exposure. P-CTX-1 inhibited axonal regrowth from axotomized peripheral neurons in culture. P-CTX-1 exposure reduced motor function in mice within the first two weeks of exposure before returning to baseline levels. These pre-exposed animals exhibited delayed sensory and motor functional recovery, and irreversible motor deficits after peripheral nerve injury in which formation of functional synapses was impaired. These findings are consistent with reduced muscle function, as assessed by electromyography recordings. Our study provides strong evidence that the persistence of P-CTX-1 in peripheral nerves reduces the intrinsic growth capacity of peripheral neurons, resulting in delayed functional recovery after injury.

Imaging Erg and Jun transcription factor interaction in living cells using fluorescence resonance energy transfer analyses

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

Camuzeaux, Barbara; Spriet, Corentin; Heliot, Laurent

2005-07-15

Physical interactions between transcription factors play important roles in modulating gene expression. Previous in vitro studies have shown a transcriptional synergy between Erg protein, an Ets family member, and Jun/Fos heterodimer, members of the bZip family, which requires direct Erg-Jun protein interactions. Visualization of protein interactions in living cells is a new challenge in biology. For this purpose, we generated fusion proteins of Erg, Fos, and Jun with yellow and cyan fluorescent proteins, YFP and CFP, respectively. After transient expression in HeLa cells, interactions of the resulting fusion proteins were explored by fluorescence resonance energy transfer microscopy (FRET) in fixedmore » and living cells. FRET between YFP-Erg and CFP-Jun was monitored by using photobleaching FRET and fluorescence lifetime imaging microscopy. Both techniques revealed the occurrence of intermolecular FRET between YFP-Erg and CFP-Jun. This is stressed by loss of FRET with an YFP-Erg version carrying a point mutation in its ETS domain. These results provide evidence for the interaction of Erg and Jun proteins in living cells as a critical prerequisite of their transcriptional synergy, but also for the essential role of the Y371 residue, conserved in most Ets proteins, in this interaction.« less

Are automatic postural responses in patients with Parkinson's disease abnormal due to their stooped posture?

PubMed

Bloem, B R; Beckley, D J; van Dijk, J G

1999-02-01

Abnormal automatic postural responses are thought to contribute to balance impairment in Parkinson's disease. However, because postural responses are modifiable by stance, we have speculated that some postural abnormalities in patients with Parkinson's disease are secondary to their stooped stance. We have studied this assumption by assessing automatic postural responses in 30 healthy subjects who were instructed either to stand upright or to assume a typical parkinsonian posture. During both conditions, subjects received 20 serial 4 degrees 'toe-up' rotational perturbations from a supporting forceplate. We recorded short-latency (SL) and medium-latency (ML) responses from stretched gastrocnemius muscles and long-latency (LL) responses from shortened tibialis anterior muscles. We also assessed changes in the center of foot pressure (CFP) and the center of gravity (COG). The results were qualitatively compared to a previously described group of patients with Parkinson's disease who, under these circumstances, typically have large ML responses, small LL responses and insufficient voluntary postural corrections, accompanied by a slow rate of backward CFP displacement and an increased posterior COG displacement. The stooped posture resulted in unloading of medial gastrocnemius muscles and loading of tibialis anterior muscles. Onset latencies of stretch responses in gastrocnemius muscles were delayed in stooped subjects, but the onset of LL responses was markedly reduced. Amplitudes of both ML and LL responses were reduced in stooped subjects. Prestimulus COG and, to a lesser extent, CFP were shifted forwards in stooped subjects. Posterior COG displacement and the rate of backward CFP displacement were diminished in stooped subjects. Voluntary postural corrections were unchanged while standing stooped. These results indicate that some postural abnormalities of patients with Parkinson's disease (most notably the reduced LL responses) can be reproduced in healthy subjects mimicking a stooped parkinsonian posture. Other postural abnormalities (most notably the increased ML responses and insufficient voluntary responses) did not appear in stooped controls and may contribute to balance impairment in Parkinson's disease.

Phospholamban mutants compete with wild type for SERCA binding in living cells

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

Gruber, Simon J.; Haydon, Suzanne; Thomas, David D., E-mail: ddt@umn.edu

2012-04-06

Highlights: Black-Right-Pointing-Pointer PLB phosphorylation in HEK cells increased FRET between YFP-PLB and CFP-SERCA. Black-Right-Pointing-Pointer Competition: Expressing loss-of-function PLB mutants in the system decreased FRET. Black-Right-Pointing-Pointer The FRET assay could screen potential therapeutic PLB mutants to activate SERCA. -- Abstract: We have used fluorescent fusion proteins stably expressed in HEK cells to detect directly the interaction between the sarcoplasmic reticulum Ca-ATPase (SERCA) and phospholamban (PLB) in living cells, in order to design PLB mutants for gene therapy. Ca{sup 2+} cycling in muscle cells depends strongly on SERCA. Heart failure (HF), which contributes to 12% of US deaths, typically exhibits decreased SERCAmore » activity, and several potential therapies for HF aim to increase SERCA activity. We are investigating the use of LOF-PLB mutants (PLB{sub M}) as gene therapy vectors to increase SERCA activity. Active SERCA1a and WT-PLB, tagged at their N termini with fluorescent proteins (CFP and YFP), were coexpressed in stable HEK cell lines, and fluorescence resonance energy transfer (FRET) was used to detect their interaction directly. Phosphorylation of PLB, induced by forskolin, caused an increase in FRET from CFP-SERCA to YFP-PLB, indicating that SERCA inhibition can be relieved without dissociation of the complex. This suggests that a LOF mutant might bind to SERCA with sufficient affinity to complete effectively with WT-PLB, thus relieving SERCA inhibition. Therefore, we transiently expressed a series of PLB{sub M} in the CFP-SERCA/YFP-PLB cell line, and found decreased FRET, implying competition between PLB{sub M} and WT-PLB for binding to SERCA. These results establish this FRET assay as a rapid and quantitative means of screening PLB{sub M} for optimization of gene therapy to activate SERCA, as needed for gene therapy in HF.« less

CD137 is a Useful Marker for Identifying CD4+ T Cell Responses to Mycobacterium tuberculosis.

PubMed

Yan, Z-H; Zheng, X-F; Yi, L; Wang, J; Wang, X-J; Wei, P-J; Jia, H-Y; Zhou, L-J; Zhao, Y-L; Zhang, H-T

2017-05-01

Upregulation of CD137 on recently activated CD8 + T cells has been used to identify rare viral and tumour antigen-specific T cells from the peripheral blood. We aimed to evaluate the accuracy of CD137 for identifying Mycobacterium tuberculosis (Mtb)-reactive CD4 + T cells in the peripheral blood of infected individuals by flow cytometry and to investigate the characteristics of these CD137 + CD4 + T cells. We initially enrolled 31 active tuberculosis (TB) patients, 31 individuals with latent TB infection (LTBI) and 25 healthy donors. The intracellular CD137 and interferon-γ (IFN-γ) production by CD4 + T cells was simultaneously detected under unstimulated and CFP10-stimulated (culture filtrate protein 10, a Mtb-specific antigen) conditions. In unstimulated CD4 + T cells, we found that the CD137 expression in the TB group was significantly higher than that in the LTBI group. Stimulation with CFP10 largely increased the CD4 + T cell CD137 expression in both the TB and LTBI groups. After CFP10 stimulation, the frequency of CD137 + CD4 + T cells was higher than that of IFN-γ + CD4 + T cells in both the TB and LTBI groups. Most of the CFP10-activated IFN-γ-secreting cells were CD137-positive, but only a small fraction of the CD137-positive cells expressed IFN-γ. An additional 20 patients with TB were enrolled to characterize the CD45RO + CCR7 + , CD45RO + CCR7 - and CD45RO - subsets in the CD137 + CD4 + T cell populations. The Mtb-specific CD137 + CD4 + T cells were mainly identified as having an effector memory phenotype. In conclusion, CD137 is a useful marker that can be used for identifying Mtb-reactive CD4 + T cells by flow cytometry. © 2017 The Foundation for the Scandinavian Journal of Immunology.

Characterization and Catalytic Upgrading of Aqueous Stream Carbon from Catalytic Fast Pyrolysis of Biomass

DOE PAGES

Starace, Anne K.; Black, Brenna A.; Lee, David D.; ...

2017-10-23

Catalytic fast pyrolysis (CFP) of biomass produces a liquid product consisting of organic and aqueous streams. The organic stream is typically slated for hydrotreating to produce hydrocarbon biofuels, while the aqueous stream is considered a waste stream, resulting in the loss of residual biogenic carbon. Here, we report the detailed characterization and catalytic conversion of a CFP wastewater stream with the ultimate aim to improve overall biomass utilization within a thermochemical biorefinery. An aqueous stream derived from CFP of beech wood was comprehensively characterized, quantifying 53 organic compounds to a total of 17% organics. The most abundant classes of compoundsmore » are acids, aldehydes, and alcohols. The most abundant components identified in the aqueous stream were C1-C2 organics, comprising 6.40% acetic acid, 2.16% methanol, and 1.84% formaldehyde on wet basis. The CFP aqueous stream was catalytically upgraded to olefins and aromatic hydrocarbons using a Ga/HZSM-5 catalyst at 500 degrees C. When the conversion yield of the upgraded products was measured with fresh, active catalyst, 33% of the carbon in the aqueous stream was recovered as aromatic hydrocarbons and 29% as olefins. The majority of the experiments were conducted using a molecular beam mass spectrometer and separate GC-MS/FID experiments were used to confirm the assignments and quantification of products with fresh excess catalyst. The recovered 62% carbon in the form of olefins and aromatics can be used to make coproducts and/or fuels potentially improving biorefinery economics and sustainability. Spent catalysts were collected after exposure to varying amounts of the feed, and were characterized using multipoint-Brunauer-Emmett-Teller (BET) adsorption, ammonia temperature programmed desorption (TPD), and thermogravimetric analysis (TGA) to monitor deactivation of Ga/HZSM-5. These characterization data revealed that deactivation was caused by coke deposits, which blocked access to active sites of the catalyst and spent catalysts regained total activity after regeneration.« less

Characterization and Catalytic Upgrading of Aqueous Stream Carbon from Catalytic Fast Pyrolysis of Biomass

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

Starace, Anne K.; Black, Brenna A.; Lee, David D.

Catalytic fast pyrolysis (CFP) of biomass produces a liquid product consisting of organic and aqueous streams. The organic stream is typically slated for hydrotreating to produce hydrocarbon biofuels, while the aqueous stream is considered a waste stream, resulting in the loss of residual biogenic carbon. Here, we report the detailed characterization and catalytic conversion of a CFP wastewater stream with the ultimate aim to improve overall biomass utilization within a thermochemical biorefinery. An aqueous stream derived from CFP of beech wood was comprehensively characterized, quantifying 53 organic compounds to a total of 17% organics. The most abundant classes of compoundsmore » are acids, aldehydes, and alcohols. The most abundant components identified in the aqueous stream were C1-C2 organics, comprising 6.40% acetic acid, 2.16% methanol, and 1.84% formaldehyde on wet basis. The CFP aqueous stream was catalytically upgraded to olefins and aromatic hydrocarbons using a Ga/HZSM-5 catalyst at 500 degrees C. When the conversion yield of the upgraded products was measured with fresh, active catalyst, 33% of the carbon in the aqueous stream was recovered as aromatic hydrocarbons and 29% as olefins. The majority of the experiments were conducted using a molecular beam mass spectrometer and separate GC-MS/FID experiments were used to confirm the assignments and quantification of products with fresh excess catalyst. The recovered 62% carbon in the form of olefins and aromatics can be used to make coproducts and/or fuels potentially improving biorefinery economics and sustainability. Spent catalysts were collected after exposure to varying amounts of the feed, and were characterized using multipoint-Brunauer-Emmett-Teller (BET) adsorption, ammonia temperature programmed desorption (TPD), and thermogravimetric analysis (TGA) to monitor deactivation of Ga/HZSM-5. These characterization data revealed that deactivation was caused by coke deposits, which blocked access to active sites of the catalyst and spent catalysts regained total activity after regeneration.« less

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

Volker, Arno; Hunter, Alan

Anisotropic materials are being used increasingly in high performance industrial applications, particularly in the aeronautical and nuclear industries. Some important examples of these materials are composites, single-crystal and heavy-grained metals. Ultrasonic array imaging in these materials requires exact knowledge of the anisotropic material properties. Without this information, the images can be adversely affected, causing a reduction in defect detection and characterization performance. The imaging operation can be formulated in two consecutive and reciprocal focusing steps, i.e., focusing the sources and then focusing the receivers. Applying just one of these focusing steps yields an interesting intermediate domain. The resulting common focusmore » point gather (CFP-gather) can be interpreted to determine the propagation operator. After focusing the sources, the observed travel-time in the CFP-gather describes the propagation from the focus point to the receivers. If the correct propagation operator is used, the measured travel-times should be the same as the time-reversed focusing operator due to reciprocity. This makes it possible to iteratively update the focusing operator using the data only and allows the material to be imaged without explicit knowledge of the anisotropic material parameters. Furthermore, the determined propagation operator can also be used to invert for the anisotropic medium parameters. This paper details the proposed technique and demonstrates its use on simulated array data from a specimen of Inconel single-crystal alloy commonly used in the aeronautical and nuclear industries.« less

Managing clinical integration in integrated delivery systems: a framework for action.

PubMed

Young, D W; Barrett, D

1997-01-01

An integrated delivery system (IDS) in healthcare must coordinate patient care across multiple functions, activities, and operating units. To achieve this clinical integration, senior management confronts many challenges. This paper uses a cross-functional-process (CFP) framework to discuss these challenges. There are ten CFPs that fall into three categories: planning processes (strategy formulation, program adaptation, budget formulation), organizational processes (authority and influence, client management, conflict resolution, motivation, and cultural maintenance), and measurement and reporting processes (financial and programmatic). Each process typically spans several functional units. Senior management must consider how to improve both the functioning of each CFP, as well as its "fit" with the other nine. The result can be greater clinical integration, improved cost management, and more coordinated care for enrollees.

Real-time detection of caspase-2 activation in a single living HeLa cell during cisplatin-induced apoptosis

NASA Astrophysics Data System (ADS)

Lin, Juqiang; Zhang, Zhihong; Yang, Jie; Zeng, Shaoqun; Liu, Bifeng; Luo, Qingming

2006-03-01

Caspase-2 is important for the mitochondrial apoptotic pathway, however, the mechanism by which caspase-2 executes apoptosis remains obscure. We carry out the first measurements of the dynamics of caspase-2 activation in a single living cell by a FRET (fluorescence resonance energy transfer) probe. Two FRET probes are constructed that each encoded a CRS (caspase-2 or caspase-3 recognition site) fused with a cyan fluorescent protein (CFP) and a red fluorescent protein (DsRed) (CFP-CRS-DsRed). Using these probes, we found that during cisplatin-induced apoptosis, caspase-2 activation occurred more slowly than did activation of caspase-3; additionally, caspase-2 activation was initiated much earlier than that of caspase-3.

Identification and functional analysis of two Golgi-localized UDP-galactofuranose transporters with overlapping functions in Aspergillus niger.

PubMed

Park, Joohae; Tefsen, Boris; Heemskerk, Marc J; Lagendijk, Ellen L; van den Hondel, Cees A M J J; van Die, Irma; Ram, Arthur F J

2015-11-02

Galactofuranose (Galf)-containing glycoconjugates are present in numerous microbes, including filamentous fungi where they are important for morphology, virulence and maintaining cell wall integrity. The incorporation of Galf-residues into galactomannan, galactomannoproteins and glycolipids is carried out by Golgi-localized Galf transferases. The nucleotide sugar donor used by these transferases (UDP-Galf) is produced in the cytoplasm and has to be transported to the lumen of the Golgi by a dedicated nucleotide sugar transporter. Based on homology with recently identified UDP-Galf-transporters in A. fumigatus and A. nidulans, two putative UDP-Galf-transporters in A. niger were found. Their function and localization was determined by gene deletions and GFP-tagging studies, respectively. The two putative UDP-Galf-transporters in A. niger are homologous to each other and are predicted to contain eleven transmembrane domains (UgtA) or ten transmembrane domains (UgtB) due to a reduced length of the C-terminal part of the UgtB protein. The presence of two putative UDP-Galf-transporters in the genome was not unique for A. niger. From the twenty Aspergillus species analysed, nine species contained two additional putative UDP-Galf-transporters. Three of the nine species were outside the Aspergillus section nigri, indication an early duplication of UDP-Galf-transporters and subsequent loss of the UgtB copy in several aspergilli. Deletion analysis of the single and double mutants in A. niger indicated that the two putative UDP-Galf-transporters (named UgtA and UgtB) have a redundant function in UDP-Galf-transport as only the double mutant displayed a Galf-negative phenotype. The Galf-negative phenotype of the double mutant could be complemented by expressing either CFP-UgtA or CFP-UgtB fusion proteins from their endogenous promoters, indicating that both CFP-tagged proteins are functional. Both Ugt proteins co-localize with each other as well as with the GDP-mannose nucleotide transporter, as was demonstrated by fluorescence microscopy, thereby confirming their predicted localization in the Golgi. A. niger contains two genes encoding UDP-Galf-transporters. Deletion and localization studies indicate that UgtA and UgtB have redundant functions in the biosynthesis of Galf-containing glycoconjugates.

Tectus niloticus (Tegulidae, Gastropod) as a Novel Vector of Ciguatera Poisoning: Detection of Pacific Ciguatoxins in Toxic Samples from Nuku Hiva Island (French Polynesia)

PubMed Central

Viallon, Jérôme; Gatti, Clémence Mahana iti; Vandersea, Mark W.; Tester, Patricia A.; Litaker, R. Wayne; Chinain, Mireille

2017-01-01

Ciguatera fish poisoning (CFP) is a foodborne disease caused by the consumption of seafood (fish and marine invertebrates) contaminated with ciguatoxins (CTXs) produced by dinoflagellates in the genus Gambierdiscus. The report of a CFP-like mass-poisoning outbreak following the consumption of Tectus niloticus (Tegulidae, Gastropod) from Anaho Bay on Nuku Hiva Island (Marquesas archipelago, French Polynesia) prompted field investigations to assess the presence of CTXs in T. niloticus. Samples were collected from Anaho Bay, 1, 6 and 28 months after this poisoning outbreak, as well as in Taiohae and Taipivai bays. Toxicity analysis using the neuroblastoma cell-based assay (CBA-N2a) detected the presence of CTXs only in Anaho Bay T. niloticus samples. This is consistent with qPCR results on window screen samples indicating the presence of Gambierdiscus communities dominated by the species G. polynesiensis in Anaho Bay. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses revealed that P-CTX-3B was the major congener, followed by P-CTX-3C, P-CTX-4A and P-CTX-4B in toxic samples. Between July 2014 and November 2016, toxin content in T. niloticus progressively decreased, but was consistently above the safety limit recommended for human consumption. This study confirms for the first time T. niloticus as a novel vector of CFP in French Polynesia. PMID:29267222

Trade-off between carbon emission and effluent quality of activated sludge processes under seasonal variations of wastewater temperature and mean cell retention time.

PubMed

Guo, Jingbo; Fu, Xin; Andrés Baquero, G; Sobhani, Reza; Nolasco, Daniel A; Rosso, Diego

2016-03-15

Over the seasonal cycles, the mean cell retention time (MCRT) of the activated sludge process is varied to compensate the wastewater temperature variations. The effects of these variations on the carbon footprint (CFP) and effluent quality index (EQI) of a conventional activated sludge (CAS) process and a nitrification/denitrification (NDN) process were quantified. The carbon emission included both biogenic and non-biogenic carbon. Carbon emissions of wasted biosolids management were also addressed. Our results confirmed that the effluent quality indicated by EQI was not necessarily improved by increasing MCRT. Higher MCRT increased the carbon emission and reduced excess sludge production, which decreased the potential for biogas energy recovery. The NDN process was preferable to the CAS process from the perspective of effluent quality. This consideration extended to the whole plant CFP if the N2O emitted during NDN was limited ([N2O]<1% [NH4(+)]removed) as the carbon emission per unit effluent quality achieved by NDN process is less than that of the CAS process. By putting forward carbon emission intensity (γ) derived from CFP and EQI, our work provides a quantitative tool for decision makers evaluating process alternatives when there is a trade-off between carbon emission and effluent quality. Copyright © 2015 Elsevier B.V. All rights reserved.

Tectus niloticus (Tegulidae, Gastropod) as a Novel Vector of Ciguatera Poisoning: Detection of Pacific Ciguatoxins in Toxic Samples from Nuku Hiva Island (French Polynesia).

PubMed

Darius, Hélène Taiana; Roué, Mélanie; Sibat, Manoella; Viallon, Jérôme; Gatti, Clémence Mahana Iti; Vandersea, Mark W; Tester, Patricia A; Litaker, R Wayne; Amzil, Zouher; Hess, Philipp; Chinain, Mireille

2017-12-21

Ciguatera fish poisoning (CFP) is a foodborne disease caused by the consumption of seafood (fish and marine invertebrates) contaminated with ciguatoxins (CTXs) produced by dinoflagellates in the genus Gambierdiscus . The report of a CFP-like mass-poisoning outbreak following the consumption of Tectus niloticus (Tegulidae, Gastropod) from Anaho Bay on Nuku Hiva Island (Marquesas archipelago, French Polynesia) prompted field investigations to assess the presence of CTXs in T. niloticus . Samples were collected from Anaho Bay, 1, 6 and 28 months after this poisoning outbreak, as well as in Taiohae and Taipivai bays. Toxicity analysis using the neuroblastoma cell-based assay (CBA-N2a) detected the presence of CTXs only in Anaho Bay T. niloticus samples. This is consistent with qPCR results on window screen samples indicating the presence of Gambierdiscus communities dominated by the species G. polynesiensis in Anaho Bay. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses revealed that P-CTX-3B was the major congener, followed by P-CTX-3C, P-CTX-4A and P-CTX-4B in toxic samples. Between July 2014 and November 2016, toxin content in T. niloticus progressively decreased, but was consistently above the safety limit recommended for human consumption. This study confirms for the first time T. niloticus as a novel vector of CFP in French Polynesia.

Investigation of Coronal Leakage of Root Fillings after Smear Layer Removal with EDTA or Er,Cr:YSGG Laser through Capillary Flow Porometry

PubMed Central

Vergauwen, Tom Edgard Maria; Michiels, Rafaël; Torbeyns, Dries; Meire, Maarten; De Moor, Roeland Jozef Gentil

2014-01-01

No studies have been performed evaluating the marginal seal of root fillings after direct exposure of root canal (RC) walls to Er,Cr:YSGG laser irradiation. Therefore, 75 root filled teeth (5 × 15–cold lateral condensation) were analyzed for through-and-through leakage (TTL) using capillary flow porometry (CFP). The cleaning protocol determined the experimental groups: (1) irrigation with NaOCl 2.5% and EDTA 17% or standard protocol (SP), (2) SP + Er,Cr:YSGG lasing (dried RC), (3) NaOCl 2.5% + Er,Cr:YSGG lasing (dried RC), (4) SP + Er,Cr:YSGG lasing (wet RC), and (5) NaOCl 2.5% + Er,Cr:YSGG lasing (wet RC). Groups 6 to 10 consisted of the same filled teeth with resected apices. Resection was performed after the first CFP measurement. CFP was used to assess minimum, mean flow, and maximum pore diameters after 48 h. Statistics were performed using nonparametric tests (P > 0.05). Additional three roots per group were submitted to SEM of the RC walls. TTL was observed in all groups without statistically significant differences between the different groups for minimum, mean, and maximum pore diameter (P > 0.05). In this study, the use of EDTA and/or Er,Cr:YSGG laser did not reduce through-and-through leakage in nonresected and resected roots. PMID:24696685

Imaging of Ras/Raf activity induced by low energy laser irradiation in living cell using FRET

NASA Astrophysics Data System (ADS)

Wang, Fang; Chen, Tong-Sheng; Xing, Da

2005-01-01

Ras/Raf signaling pathway is an important signaling pathway that governs cell proliferation, differential and apoptosis. Low-energy laser irradiation (LELI) was found to modulate various processes. Generally, cell proliferation is induced by low doses LELI and apoptosis is induced by high doses LELI. Mechanism of biological effect of LELI has not been clear. Recently, activation of MEK (mitogen-activated protein kinase) and ERK (extracellular-signal-regulated kinase), which are downstream protein kinases of Ras/Raf, are observed during LELI-induced cell proliferation by immunoprecipitation and western blot analysis. RaichuRas reporter consisting of fusions of H-ras, the Ras-binding domain of Raf (RafRBD), a cyan fluorescent protein (CFP) and a yellow fluorescent protein (YFP). Therefore, intramolecular binding of GTP-Ras to RafRBD brings CFP close to YFP and increases FRET between CFP and YFP. Human lung adenocarcinoma cell line (ASTC-a-1) was transfected with the plasmid (pRaichuRas) and then treated with LELI at dose of 60J/cm2. Effect of LELI on Ras/Raf in physiological condition of living cells was observed by fluorescence resonance energy transfer (FRET) technique during lung adenocarcinoma cell apoptosis induced by high dose (60J/cm2) LELI. Experimental results showed that after high dose LELI treatment, the binding of Ras and Raf decreases obviously, Ras/Raf signaling pathway deregulates and cell apoptosis occurs.

Reversion and conversion of Mycobacterium tuberculosis IFN-gamma ELISpot results during anti-tuberculous treatment in HIV-infected children.

PubMed

Connell, Tom G; Davies, Mary-Ann; Johannisen, Christine; Wood, Kathryn; Pienaar, Sandy; Wilkinson, Katalin A; Wilkinson, Robert J; Zar, Heather J; Beatty, David; Nicol, Mark P; Curtis, Nigel; Eley, Brian

2010-05-27

Recent interest has focused on the potential use of serial interferon gamma (IFN-gamma) release assay (IGRA) measurements to assess the response to anti-tuberculous (TB) treatment. The kinetics of IFN-gamma responses to Mycobacterium tuberculosis (MTB) antigens in HIV-infected children during treatment have not however been previously investigated. IFN-gamma responses to the MTB antigens, ESAT-6, CFP-10 and PPD were measured by an enzyme-linked immunospot assay (IFN-gamma ELISpot) at presentation and at one, two and six months after starting anti-tuberculous treatment in HIV-infected children with definite or probable TB. Responses at different time points were compared using a Mann-Whitney U test with paired data analysed using the Wilcoxon signed rank test. A Fisher's exact or Chi-squared test was used to compare proportions when test results were analysed as dichotomous outcomes. Of 102 children with suspected TB, 22 (21%) had definite TB and 24 (23%) probable TB. At least one follow up IFN-gamma ELISpot assay result was available for 31 (67%) of the 46 children. In children with definite or probable TB in whom the IFN-gamma ELISpot assay result was positive at presentation, anti-tuberculous treatment was accompanied by a significant decrease in both the magnitude of the IFN-gamma response to individual or combined MTB-specific antigens (ESAT-6 median 110 SFCs/106 PBMC (IQR 65-305) at presentation vs. 15 (10-115) at six months, p = 0.04; CFP-10 177 (48-508) vs. 20 (5-165), p = 0.004, ESAT-6 or CFP-10 median 250 SFCs/106 PBMC (IQR 94-508) vs. 25 (10-165), p = 0.004) and in the proportion of children with a positive IFN-gamma ELISpot assay (Fisher's exact test: ESAT-6 15/0 vs 5/11, p = 0.0002, CFP-10 22/0 vs 8/17, p = 0.0001, ESAT-6 or CFP-10 22/0 vs. 9/17, p= 0.002). However almost half of the children had a positive IFN-gamma ELISpot assay after six months of anti-tuberculous treatment. In addition, there was conversion of the IFN-gamma ELISpot assay result during anti-tuberculous therapy in six of 12 children in whom the initial IFN-gamma ELISpot assay was negative. In HIV-infected children with definite or probable TB, anti-tuberculosis treatment is accompanied by a reduction in the magnitude of the IFN-gamma ELISpot response to MTB-antigens. However, serial IFN-gamma ELISpot measurements appear to have limited clinical utility in assessing a successful response to anti-tuberculous treatment in HIV infected children.

Characterization factors for water footprint considering the scarcity of green and blue water sources

NASA Astrophysics Data System (ADS)

Oki, T.; Kondo, T.; Pokhrel, Y. N.; Hanasaki, N.

2011-12-01

The original concept of virtual water trade was invented to illustrate how much water demand can be reduced by importing food products (Allan 1996), and expanded for meat products and industrial products (Oki and Kanae, 2004). However, there was a confusion between "virtual trade of water" (original) and "trade of virtual water" (misinterpretation but widely accepted), and "virtual water" has been recognized as how much water was used to produce the commodity. Then, the concept has some analogy to carbon footprint (CFP) which is an indicator of total emission of greenhouse gases, and nowadays called water footprint (WFP, Hoekstra, 2004). However, WFP itself is just an inventory of water usages under the framework of life cycle assessment (LCA), and the volume of WFP does not necessary reflect the environmental impacts of water usages because consumptive water use of 100 liter from ground water in arid regions just before rainy season would have more environmental impacts than consumptive water use of 100 liter from rain water in humid regions during snow melt season. In the case of CFP, the emissions of five greenhouse gases except for CO2 were converted into CO2 equivalent volumes by considering the sensitivity for the global warming potential, and summed up into CFP. Here, we propose a new idea objectively determining the weights (characterization factors) for blue water usages, such as from river and ground water, to be converted into green water equivalent in each region and time. The weights are inversely proportional to the area required to obtain the same amount of green water, and water balance model can provide the basic information. The new concept was applied to the WFP of Japan through the imports of major crops. As an inventory, WFP was 15.5 km3/y of rain water, 2.2 km3/y of river water, and 2.0 km3/y of non-renewable and non-local water (NRNL water) for year 2000, however, considering the proposed characterization factors in each region (0.5 x 0.5 degree global grids) based on annual water balances, WFPs become 8.1 km3/y of river water and 22.0 km3/y of NRNL water. This new concept can be expanded into an idea of "global mean green water equivalent" volume of WFP.

Skin test performed with highly purified Mycobacterium tuberculosis recombinant protein triggers tuberculin shock in infected guinea pigs.

PubMed

Reece, Stephen T; Stride, Nicole; Ovendale, Pamela; Reed, Steven G; Campos-Neto, Antonio

2005-06-01

Tuberculin shock due to inoculation of Mycobacterium tuberculosis antigens in patients with tuberculosis is a serious syndrome originally described over 100 years ago by Robert Koch. Here, we present experimental evidence that a single M. tuberculosis recombinant protein, CFP-10, triggers this syndrome. Intradermal inoculation of CFP-10 elicits in M. tuberculosis-infected mice high levels of serum tumor necrosis factor alpha and causes tuberculin shock in infected guinea pigs characterized by hypothermia and death within 6 to 48 h after the antigen inoculation. Autopsies of these animals revealed intense polycythemia and hemorrhagic patches in the lung parenchyma, a pathological observation consistent with tuberculin shock. These results point to the possible occurrence of tuberculin shock in sensitive individuals inoculated with highly purified M. tuberculosis recombinant proteins as vaccine candidates or skin test reagents.

Genomic data reveal ancient microendemism in forest scorpions across the California Floristic Province.

PubMed

Bryson, Robert W; Savary, Warren E; Zellmer, Amanda J; Bury, R Bruce; McCormack, John E

2016-08-01

The California Floristic Province (CFP) in western North America is a globally significant biodiversity hotspot. Elucidating patterns of endemism and the historical drivers of this diversity has been an important challenge of comparative phylogeography for over two decades. We generated phylogenomic data using ddRADseq to examine genetic structure in Uroctonus forest scorpions, an ecologically restricted and dispersal-limited organism widely distributed across the CFP north to the Columbia River. We coupled our genetic data with species distribution models (SDMs) to determine climatically suitable areas for Uroctonus both now and during the Last Glacial Maximum. Based on our analyses, Uroctonus is composed of two major genetic groups that likely diverged over 2 million years ago. Each of these groups itself contains numerous genetic groups that reveal a pattern of vicariance and microendemism across the CFP. Migration rates among these populations are low. SDMs suggest forest scorpion habitat has remained relatively stable over the last 21 000 years, consistent with the genetic data. Our results suggest tectonic plate rafting, mountain uplift, river drainage formation and climate-induced habitat fragmentation have all likely played a role in the diversification of Uroctonus. The intricate pattern of genetic fragmentation revealed across a temporal continuum highlights the potential of low-dispersing species to shed light on small-scale patterns of biodiversity and the underlying processes that have generated this diversity in biodiversity hotspots. © 2016 John Wiley & Sons Ltd.

Carbon Footprint of Biofuel Sugarcane Produced in Mineral and Organic Soils in Florida

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

Izursa, Jose-Luis; Hanlon, Edward; Amponsah, Nana

2013-02-06

Ethanol produced from sugarcane is an existing and accessible form of renewable energy. In this study, we applied the Life Cycle Assessment (LCA) approach to estimate the Carbon Footprint (CFP) of biofuel sugarcane produced on mineral (sandy) and organic (muck) soils in Florida. CFP was estimated from greenhouse gas (GHG) emissions (CO2, CH4, and N2O) during the biofuel sugarcane cultivation. The data for the energy (fossil fuels and electricity), equipment, and chemical fertilizers were taken from enterprise budgets prepared by the University of Florida based on surveys and interviews obtained from local growers during the cropping years 2007/2008 and 2009/2010more » for mineral soils and 2008/2009 for organic soils. Emissions from biomass burning and organic land use were calculated based on the IPCC guidelines. The results show that the CFP for biofuel sugarcane production is 0.04 kg CO2e kg-1y-1 when produced in mineral soils and 0.46 kg CO2e kg-1y-1 when produced in organic soils. Most of the GHG emissions from production of biofuel sugarcane in mineral soils come from equipment (33%), fertilizers (28%), and biomass burning (27%); whereas GHG emissions from production in organic soils come predominantly from the soil (93%). This difference should be considered to adopt new practices for a more sustainable farming system if biofuel feedstocks are to be considered.« less

Identification of Residues Involved in Substrate Specificity and Cytotoxicity of Two Closely Related Cutinases from Mycobacterium tuberculosis

PubMed Central

Dedieu, Luc; Serveau-Avesque, Carole; Canaan, Stéphane

2013-01-01

The enzymes belonging to the cutinase family are serine enzymes active on a large panel of substrates such as cutin, triacylglycerols, and phospholipids. In the M. tuberculosis H37Rv genome, seven genes coding for cutinase-like proteins have been identified with strong immunogenic properties suggesting a potential role as vaccine candidates. Two of these enzymes which are secreted and highly homologous, possess distinct substrates specificities. Cfp21 is a lipase and Cut4 is a phospholipase A2, which has cytotoxic effects on macrophages. Structural overlay of their three-dimensional models allowed us to identify three areas involved in the substrate binding process and to shed light on this substrate specificity. By site-directed mutagenesis, residues present in these Cfp21 areas were replaced by residues occurring in Cut4 at the same location. Three mutants acquired phospholipase A1 and A2 activities and the lipase activities of two mutants were 3 and 15 fold greater than the Cfp21 wild type enzyme. In addition, contrary to mutants with enhanced lipase activity, mutants that acquired phospholipase B activities induced macrophage lysis as efficiently as Cut4 which emphasizes the relationship between apparent phospholipase A2 activity and cytotoxicity. Modification of areas involved in substrate specificity, generate recombinant enzymes with higher activity, which may be more immunogenic than the wild type enzymes and could therefore constitute promising candidates for antituberculous vaccine production. PMID:23843969

Identification of residues involved in substrate specificity and cytotoxicity of two closely related cutinases from Mycobacterium tuberculosis.

PubMed

Dedieu, Luc; Serveau-Avesque, Carole; Canaan, Stéphane

2013-01-01

The enzymes belonging to the cutinase family are serine enzymes active on a large panel of substrates such as cutin, triacylglycerols, and phospholipids. In the M. tuberculosis H37Rv genome, seven genes coding for cutinase-like proteins have been identified with strong immunogenic properties suggesting a potential role as vaccine candidates. Two of these enzymes which are secreted and highly homologous, possess distinct substrates specificities. Cfp21 is a lipase and Cut4 is a phospholipase A2, which has cytotoxic effects on macrophages. Structural overlay of their three-dimensional models allowed us to identify three areas involved in the substrate binding process and to shed light on this substrate specificity. By site-directed mutagenesis, residues present in these Cfp21 areas were replaced by residues occurring in Cut4 at the same location. Three mutants acquired phospholipase A1 and A2 activities and the lipase activities of two mutants were 3 and 15 fold greater than the Cfp21 wild type enzyme. In addition, contrary to mutants with enhanced lipase activity, mutants that acquired phospholipase B activities induced macrophage lysis as efficiently as Cut4 which emphasizes the relationship between apparent phospholipase A2 activity and cytotoxicity. Modification of areas involved in substrate specificity, generate recombinant enzymes with higher activity, which may be more immunogenic than the wild type enzymes and could therefore constitute promising candidates for antituberculous vaccine production.

Highly Sensitive and Practical Fluorescent Sandwich ELISA for Ciguatoxins.

PubMed

Tsumuraya, Takeshi; Sato, Takeshi; Hirama, Masahiro; Fujii, Ikuo

2018-05-29

Ciguatera fish poisoning (CFP) caused by the consumption of fish that have accumulated ciguatoxins (CTXs) affects more than 50000 people annually. The spread of CFP causes enormous damage to public health, fishery resources, and the economies of tropical and subtropical endemic regions. The difficulty in avoiding CFP arises from the lack of sensitive and reliable analytical methods for the detection and quantification of CTXs in contaminated fish, along with the normal appearance, smell, and taste of fish contaminated with the causative toxins. Thus, an accurate, sensitive, routine, and portable detection method for CTXs is urgently required. We have successfully developed a highly sensitive fluorescent sandwich ELISA, which can detect, differentiate, and quantify four major CTX congeners (CTX1B, CTX3C, 51-hydroxyCTX3C, and 54-deoxyCTX1B) with a detection limit of less than 1 pg/mL. The ELISA protocol, using one microtiter plate coated with two mAbs (10C9 and 3G8), and ALP-linked 8H4, can detect any of the four CTX congeners in a single operation. CTX1B spiked into fish at the FDA guidance level of 0.01 ppb CTX1B equivalent toxicity in fish from Pacific regions was also proven to be reliably detected by this ELISA. Furthermore, the efficiency of extraction/purification procedures and the matrix effect of contaminants in fish were evaluated in detail, since pretreatment and matrix effects are critical for ELISA analysis.

Specificity of the Tuberculin Skin Test Is Modified by Use of a Protein Cocktail Containing ESAT-6 and CFP-10 in Cattle Naturally Infected with Mycobacterium bovis

PubMed Central

Flores-Villalva, S.; Suárez-Güemes, F.; Espitia, C.; Whelan, A. O.; Vordermeier, M.

2012-01-01

The mycobacterial immunodominant ESAT-6 and CFP-10 antigens are strongly recognizable in tuberculosis-infected cattle, and they do not elicit a response in cattle without infection. In addition, they are absent in most environmental mycobacterial species, and therefore, their use can be an alternative to purified protein derivative (PPD) tuberculin in the development of a more specific skin diagnostic test in cattle. The aim of the current study was to assess the potential of an ESAT-6 and CFP-10 (E6-C10) protein cocktail in a skin test format in naturally tuberculosis-infected and paratuberculosis-infected cattle. We also included MPB83 as a third component in one of the protein cocktail preparations. The protein cocktail was tested at different dose concentrations (5, 10, and 15 μg per protein). The best skin response to the E6-C10 protein cocktail was obtained with 10 μg. Subsequently, this concentration was tested in 2 herds with high and low bovine tuberculosis prevalence, the latter with paratuberculosis coinfection. Our data show that the E6-C10 cocktail allows identification of an important proportion of animals that PPDB is not able to recognize, especially in low-prevalence herds. The protein cocktail did not induce reactions in tuberculosis-free cattle or in paratuberculosis-infected cattle. Addition of MPB83 to the protein cocktail did not make any difference in the skin reaction. PMID:22419675

Specificity of the tuberculin skin test is modified by use of a protein cocktail containing ESAT-6 and CFP-10 in cattle naturally infected with Mycobacterium bovis.

PubMed

Flores-Villalva, S; Suárez-Güemes, F; Espitia, C; Whelan, A O; Vordermeier, M; Gutiérrez-Pabello, J A

2012-05-01

The mycobacterial immunodominant ESAT-6 and CFP-10 antigens are strongly recognizable in tuberculosis-infected cattle, and they do not elicit a response in cattle without infection. In addition, they are absent in most environmental mycobacterial species, and therefore, their use can be an alternative to purified protein derivative (PPD) tuberculin in the development of a more specific skin diagnostic test in cattle. The aim of the current study was to assess the potential of an ESAT-6 and CFP-10 (E6-C10) protein cocktail in a skin test format in naturally tuberculosis-infected and paratuberculosis-infected cattle. We also included MPB83 as a third component in one of the protein cocktail preparations. The protein cocktail was tested at different dose concentrations (5, 10, and 15 μg per protein). The best skin response to the E6-C10 protein cocktail was obtained with 10 μg. Subsequently, this concentration was tested in 2 herds with high and low bovine tuberculosis prevalence, the latter with paratuberculosis coinfection. Our data show that the E6-C10 cocktail allows identification of an important proportion of animals that PPDB is not able to recognize, especially in low-prevalence herds. The protein cocktail did not induce reactions in tuberculosis-free cattle or in paratuberculosis-infected cattle. Addition of MPB83 to the protein cocktail did not make any difference in the skin reaction.

Integrating gene and protein expression data with genome-scale metabolic networks to infer functional pathways.

PubMed

Pey, Jon; Valgepea, Kaspar; Rubio, Angel; Beasley, John E; Planes, Francisco J

2013-12-08

The study of cellular metabolism in the context of high-throughput -omics data has allowed us to decipher novel mechanisms of importance in biotechnology and health. To continue with this progress, it is essential to efficiently integrate experimental data into metabolic modeling. We present here an in-silico framework to infer relevant metabolic pathways for a particular phenotype under study based on its gene/protein expression data. This framework is based on the Carbon Flux Path (CFP) approach, a mixed-integer linear program that expands classical path finding techniques by considering additional biophysical constraints. In particular, the objective function of the CFP approach is amended to account for gene/protein expression data and influence obtained paths. This approach is termed integrative Carbon Flux Path (iCFP). We show that gene/protein expression data also influences the stoichiometric balancing of CFPs, which provides a more accurate picture of active metabolic pathways. This is illustrated in both a theoretical and real scenario. Finally, we apply this approach to find novel pathways relevant in the regulation of acetate overflow metabolism in Escherichia coli. As a result, several targets which could be relevant for better understanding of the phenomenon leading to impaired acetate overflow are proposed. A novel mathematical framework that determines functional pathways based on gene/protein expression data is presented and validated. We show that our approach is able to provide new insights into complex biological scenarios such as acetate overflow in Escherichia coli.

Subcellular targeting and interactions among the Potato virus X TGB proteins.

PubMed

Samuels, Timmy D; Ju, Ho-Jong; Ye, Chang-Ming; Motes, Christy M; Blancaflor, Elison B; Verchot-Lubicz, Jeanmarie

2007-10-25

Potato virus X (PVX) encodes three proteins named TGBp1, TGBp2, and TGBp3 which are required for virus cell-to-cell movement. To determine whether PVX TGB proteins interact during virus cell-cell movement, GFP was fused to each TGB coding sequence within the viral genome. Confocal microscopy was used to study subcellular accumulation of each protein in virus-infected plants and protoplasts. GFP:TGBp2 and TGBp3:GFP were both seen in the ER, ER-associated granular vesicles, and perinuclear X-bodies suggesting that these proteins interact in the same subdomains of the endomembrane network. When plasmids expressing CFP:TGBp2 and TGBp3:GFP were co-delivered to tobacco leaf epidermal cells, the fluorescent signals overlapped in ER-associated granular vesicles indicating that these proteins colocalize in this subcellular compartment. GFP:TGBp1 was seen in the nucleus, cytoplasm, rod-like inclusion bodies, and in punctate sites embedded in the cell wall. The puncta were reminiscent of previous reports showing viral proteins in plasmodesmata. Experiments using CFP:TGBp1 and YFP:TGBp2 or TGBp3:GFP showed CFP:TGBp1 remained in the cytoplasm surrounding the endomembrane network. There was no evidence that the granular vesicles contained TGBp1. Yeast two hybrid experiments showed TGBp1 self associates but failed to detect interactions between TGBp1 and TGBp2 or TGBp3. These experiments indicate that the PVX TGB proteins have complex subcellular accumulation patterns and likely cooperate across subcellular compartments to promote virus infection.

Mechanism study of low-energy laser irradiation-induced lung adenocarcinoma cell proliferation by FRET in living cell

NASA Astrophysics Data System (ADS)

Wang, Fang; Chen, Xiao-Chuan; Xing, Da

2004-07-01

Low-energy laser irradiation (LELI) has been shown to promote cell proliferation in various cell types, yet the mechanism of which has not been fully clarified. The Ras/Raf/MEK (mitogen-activated protein kinase)ERK kinase)/ERK (extracellular-signal-regulated kinase) signaling pathway is a network that govern proliferation, differentiation and cell survival. Recent studies suggested that Ras/Raf/MEK/ERK pathway is involved in the LELI-induced cell proliferation. Here, we utilized fluorescence resonance energy transfer (FRET) technique to investigate the effect of LELI on Ras/Raf signaling pathway in living cells. Raichu-Ras reporter plasmid was utilized which consisted of fusions of H-ras, the Ras-binding domain of Raf(RafRBD), a cyan fluorescent protein (CFP) and a yellow fluorescent protein (YFP), so that intramolecular binding of GTP-Ras to RafRBD brings CFP close to YFP and increases FRET between CFP and YFP. Human lung adenocarcinoma cell line (ASTC-a-1) were transfected with the plasmid (pRaichu-Ras) and then were treated by LELI. The living cell imaging showed the increase of FRET at different time points after LELI at the dose of 1.8 J/cm2, which corresponds to the Ras/Raf activation assayed by Western Blotting. Furthermore, this dose of LELI enhanced the proliferation of ASTC-a-1 cells. Taken together, these in vivo imaging data provide direct evidences with temporal or spatial resolution that Ras/Raf/MEK/ pathway plays an important role in LELI-promoted cell proliferation.

The Regional Advisory Councils: what is their potential to incorporate stakeholder knowledge into fisheries governance?

PubMed

Linke, Sebastian; Dreyer, Marion; Sellke, Piet

2011-03-01

The protection of the Baltic Sea ecosystem is exacerbated by the social, environmental and economic complexities of governing European fisheries. Increased stakeholder participation and knowledge integration are suggested to improve the EU's Common Fisheries Policy (CFP), suffering from legitimacy, credibility and compliance problems. As a result, the CFP was revised in 2002 to involve fisheries representatives, NGOs and other stakeholders through so called Regional Advisory Councils (RACs) in the policy process. We address the RAC's task to incorporate stakeholder knowledge into the EU's fisheries governance system in empirical and theoretical perspectives. Drawing on a four-stage governance concept we subsequently suggest that a basic problem is a mismatch between participation purpose (knowledge inclusion) and the governance stage at which RACs are formally positioned (evaluation of management proposals). We conclude that, if the aim is to broaden the knowledge base of fisheries management, stakeholders need to be included earlier in the governance process.

Responses to diagnostic tests for bovine tuberculosis in dairy and non-dairy cattle naturally exposed to Mycobacterium bovis in Great Britain.

PubMed

Downs, S H; Broughan, J M; Goodchild, A V; Upton, P A; Durr, P A

2016-10-01

Field surveillance of British cattle using the single intradermal comparative cervical tuberculin (SICCT) test shows a higher incidence rate of bovine tuberculosis (bTB) in dairy compared to beef herds, but a lower probability of post-mortem examination confirmed (PMC) Mycobacterium bovis infection in dairy herds. A cross-sectional study was conducted to compare animal level differences in bTB detection between dairy and non-dairy cattle in Great Britain. During the period from 2002 to 2005, 200 (41% dairy) reactors in the SICCT test (standard interpretation) were randomly selected, and 200 in-contact cattle (43% dairy) were purposively selected from bTB-infected herds. Interferon (IFN)-γ responses in blood to bovine and avian purified protein derivative (PPD), and early secretory antigen target 6 kDa and culture filtrate protein 10 (ESAT-6/CFP10), were measured. The post-mortem examination included gross pathological examination, mycobacterial culture and histopathology. The proportions of cattle positive to ESAT6/CFP10 were 26% (95% confidence interval, CI, 15-39%) in dairy reactors and 62% (95% CI 51-72%) in non-dairy reactors (P <0.001). PMC risk was 34% (95% CI 24-45%) in dairy reactors and 69% (95% CI 60-78%) in non-dairy reactors (P <0.001). The odds ratio for PMC risk in dairy reactors compared to non-dairy reactors, after controlling for bTB prevalence, herd size and SICCT test response, was 0.27 (95% CI 0.14-0.53; P <0.001). In surveillance data, adjusted animal level PMC risks were lower for dairy reactors than for beef reactors aged >2 years (P <0.001). Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

Three-Dimensional Reconstruction of Three-Way FRET Microscopy Improves Imaging of Multiple Protein-Protein Interactions.

PubMed

Scott, Brandon L; Hoppe, Adam D

2016-01-01

Fluorescence resonance energy transfer (FRET) microscopy is a powerful tool for imaging the interactions between fluorescently tagged proteins in two-dimensions. For FRET microscopy to reach its full potential, it must be able to image more than one pair of interacting molecules and image degradation from out-of-focus light must be reduced. Here we extend our previous work on the application of maximum likelihood methods to the 3-dimensional reconstruction of 3-way FRET interactions within cells. We validated the new method (3D-3Way FRET) by simulation and fluorescent protein test constructs expressed in cells. In addition, we improved the computational methods to create a 2-log reduction in computation time over our previous method (3DFSR). We applied 3D-3Way FRET to image the 3D subcellular distributions of HIV Gag assembly. Gag fused to three different FPs (CFP, YFP, and RFP), assembled into viral-like particles and created punctate FRET signals that become visible on the cell surface when 3D-3Way FRET was applied to the data. Control experiments in which YFP-Gag, RFP-Gag and free CFP were expressed, demonstrated localized FRET between YFP and RFP at sites of viral assembly that were not associated with CFP. 3D-3Way FRET provides the first approach for quantifying multiple FRET interactions while improving the 3D resolution of FRET microscopy data without introducing bias into the reconstructed estimates. This method should allow improvement of widefield, confocal and superresolution FRET microscopy data.

A novel assay to assess the effect of pharmaceutical compounds on the differentiation of podocytes.

PubMed

Kindt, Frances; Hammer, Elke; Kemnitz, Stefan; Blumenthal, Antje; Klemm, Paul; Schlüter, Rabea; Quaggin, Susan E; van den Brandt, Jens; Fuellen, Georg; Völker, Uwe; Endlich, Karlhans; Endlich, Nicole

2017-01-01

Therapeutic options for treating glomerulopathies, the main cause of chronic kidney disease, are limited. Podocyte dedifferentiation is a major event in the pathogenesis of glomerulopathies. The goal of the present study was, therefore, to develop an assay to monitor podocyte differentiation suitable for compound screening. We isolated and cultured glomeruli from transgenic mice, expressing cyan fluorescent protein (CFP) under the control of the promoter of nephrin, a marker of podocyte differentiation. Mean CFP fluorescence intensity per glomerulus (MFG) was determined by summation of all glomerular voxels from confocal z-stacks in the absence and presence of pharmaceutical compounds. In untreated cultured glomeruli, MFG remained fairly stable during the first 5 days, when foot processes were already effaced, and the level of many podocyte-specific proteins was only mildly affected, as revealed by proteomics. Between day 6 and 9, MFG decreased to almost zero. The decrease in MFG was paralleled by a decrease in CFP and nephrin expression, as determined by RT-PCR, western blots and proteomics. Puromycin aminonucleoside (PAN), which damages podocytes, concentration-dependently induced a complete loss of MFG. Dexamethasone (25 μM) and pioglitazone (10 μM) markedly attenuated the effect of 0.6 μg·mL -1 PAN on MFG. In summary, we established a novel assay to assess the effect of pharmaceutical compounds on the differentiation of podocytes in situ. Our assay is suitable for compound screening to identify drugs for the treatment of glomerulopathies. © 2016 The British Pharmacological Society.

A novel assay to assess the effect of pharmaceutical compounds on the differentiation of podocytes

PubMed Central

Kindt, Frances; Hammer, Elke; Kemnitz, Stefan; Blumenthal, Antje; Klemm, Paul; Schlüter, Rabea; Quaggin, Susan E; van den Brandt, Jens; Fuellen, Georg; Völker, Uwe; Endlich, Karlhans

2016-01-01

Background and Purpose Therapeutic options for treating glomerulopathies, the main cause of chronic kidney disease, are limited. Podocyte dedifferentiation is a major event in the pathogenesis of glomerulopathies. The goal of the present study was, therefore, to develop an assay to monitor podocyte differentiation suitable for compound screening. Experimental Approach We isolated and cultured glomeruli from transgenic mice, expressing cyan fluorescent protein (CFP) under the control of the promoter of nephrin, a marker of podocyte differentiation. Mean CFP fluorescence intensity per glomerulus (MFG) was determined by summation of all glomerular voxels from confocal z‐stacks in the absence and presence of pharmaceutical compounds. Key Results In untreated cultured glomeruli, MFG remained fairly stable during the first 5 days, when foot processes were already effaced, and the level of many podocyte‐specific proteins was only mildly affected, as revealed by proteomics. Between day 6 and 9, MFG decreased to almost zero. The decrease in MFG was paralleled by a decrease in CFP and nephrin expression, as determined by RT‐PCR, western blots and proteomics. Puromycin aminonucleoside (PAN), which damages podocytes, concentration‐dependently induced a complete loss of MFG. Dexamethasone (25 μM) and pioglitazone (10 μM) markedly attenuated the effect of 0.6 μg·mL−1 PAN on MFG. Conclusion and Implications In summary, we established a novel assay to assess the effect of pharmaceutical compounds on the differentiation of podocytes in situ. Our assay is suitable for compound screening to identify drugs for the treatment of glomerulopathies. PMID:27858997

In-vivo mapping of drusen by fundus autofluorescence and spectral-domain optical coherence tomography imaging.

PubMed

Göbel, Arno P; Fleckenstein, Monika; Heeren, Tjebo F C; Holz, Frank G; Schmitz-Valckenberg, Steffen

2016-01-01

To determine fundus autofluorescence (FAF) signal variations and corresponding microstructural alterations on spectral-domain optical coherence tomography (SD-OCT) in areas of funduscopically visible drusen associated with age-related macular degeneration (AMD). Thirty eyes from 22 patients with geographic atrophy (GA) secondary to AMD (median age 74, range 64-87 years), who had undergone retinal imaging including color fundus photography (CFP), FAF and SD-OCT (Spectralis HRA+OCT; Heidelberg Engineering GmbH, Heidelberg, Germany) were retrospectively analyzed. In each eye, at least one druse (≥ 63 μm) in the perilesional zone of GA recorded on CFP was analyzed. Relative FAF intensities and alterations in SD-OCT bands at the site of each druse were evaluated. A total of 73 drusen were analyzed, which were associated with heterogeneous corresponding alterations on FAF and SD-OCT. The FAF signal was normal, increased, decreased or not evaluable in 32 (44 %), 27 (37 %), 12 (16 %), and 2 (3 %) drusen, respectively. Focal hyperreflectivity overlying drusen was most frequently spatially confined to increased FAF (present in 9 (33 %) of 27 drusen with increased FAF). Outer nuclear layer thinning and choroidal hyperreflectivity were associated with decreased FAF (present in 7 [58 %] of 12 and 6 [50 %] of 12 drusen with decreased FAF, respectively). The appearance of soft drusen on CFP does not allow for differentiation between preserved and markedly compromised outer retinal integrity, including incipient atrophy and focal neurosensory alterations of reflectivity overlying extracellular sub-retinal pigment epithelium (RPE) deposits. Multimodal imaging reveals a broad spectrum of microstructural changes, which may reflect different stages in the evolution of drusen.

Influence of dopaminergic medication on automatic postural responses and balance impairment in Parkinson's disease.

PubMed

Bloem, B R; Beckley, D J; van Dijk, J G; Zwinderman, A H; Remler, M P; Roos, R A

1996-09-01

It is still unclear why balance impairment in Parkinson's disease (PD) often responds insufficiently to dopaminergic medication. We have studied this issue in 23 patients with idiopathic PD and 24 healthy controls. Our specific purposes were (a) to investigate the contribution of abnormal automatic postural responses to balance impairment in PD and (b) to assess the influence of dopaminergic medication on abnormal automatic postural responses and balance impairment. Standing subjects received 4 degrees "toe-up" rotational perturbations of a supporting forceplate. We bilaterally recorded posturally destabilizing medium latency (ML) responses from the stretched gastrocnemius muscles and functionally corrective long latency (LL) responses from the shortened tibialis anterior (TA) muscles. We also assessed changes in the center of foot pressure (CFP) and the center of gravity (COG). All patients were tested in the "off" and "on" phases. All controls were tested and retested after 1 h. During the off phase, we found enlarged ML amplitudes and diminished LL amplitudes in patients, together with a markedly increased posterior displacement of the COG. The abnormal ML and LL responses were partially responsible for the increased body sway in patients because the initial forward (destabilizing) displacement of the CFP was increased, while the subsequent backward displacement of the CFP (a measure of the corrective braking action of LL responses) was delayed. Abnormal late automatic or possibly more voluntary postural corrections also contributed substantially to the increased body sway. During the on phase, ML amplitudes were reduced in patients but remained increased compared with controls. LL amplitudes no longer differed between both groups due to a modest, possibly dopamine-related increase in patients and a simultaneous decrease in controls. The abnormal CFP displacement was only partially improved by dopaminergic medication. The later postural corrections were not improved at all. Consequently, the increased posterior COG displacement was not ameliorated during the on phase. We conclude that (a) a combination of abnormal automatic and perhaps more voluntary postural corrections contributes to increased body sway in PD and (b) dopaminergic medication fails to improve balance impairment in PD because early automatic postural responses are only partially corrected, while later occurring postural corrections are not improved at all. These electrophysiological results support clinical observations and suggest that nondopaminergic lesions play a significant role in the pathophysiology of postural abnormalities in PD.

Comparison of outgassing models for the landsat thematic mapper sensors

USGS Publications Warehouse

Micijevic, E.; Chander, G.

2007-01-01

The Thematic Mapper (TM) is a multi-spectral electro-optical sensor featured onboard both the Landsat 4 (L4) and Landsat 5 (L5) satellites. TM sensors have seven spectral bands with center wavelengths of approximately 0.49, 0.56, 0.66, 0.83, 1.65, 11.5 and 2.21 ??m, respectively. The visible near-infrared (VNIR) bands are located on the primary focal plane (PFP), and two short-wave infrared (SWIR) bands and the thermal infrared (TIR) band are located on the cold focal plane (CFP). The CFP bands are maintained at cryogenic temperatures of about 91 K, to reduce thermal noise effects. Due to the cold temperature, an ice film accumulates on the CFP dewar window, which introduces oscillations in SWIR and an exponential decay in TIR band responses. This process is usually monitored and characterized by the detector responses to the internal calibrator (IC) lamps and the blackbody. The ice contamination on the dewar window is an effect of the sensor outgassing in a vacuum of the space environment. Outgassing models have been developed, which are based on the thin-film optical interference phenomenon. They provide the coefficients for correction for outgassing effects for the entire mission's lifetime. While the L4 TM ceased imaging in August 1993, the L5 TM continues to operate even after more than 23 years in orbit. The process of outgassing in L5 TM is still occurring, though at a much lower rate than during early years of mission. Although the L4 and L5 TM sensors are essentially identical, they exhibit slightly different responses to the outgassing effects. The work presented in the paper summarizes the results of modeling outgassing effects in each of the sensors and provides a detailed analysis of differences among the estimated modeling parameters. For both sensors, water ice was confirmed as a reasonable candidate for contaminant material, the contaminant growth rate was found to be gradually decreasing with the time since launch, and the indications exist that some film may remain after the CFP warm-up procedures, which are periodically initiated to remove accumulated contamination. The observed difference between the models could be contributed to differences in the operational history for the sensors, the content and amount of contaminant impurities, the sensor spectral filter responses, and the internal calibrator systems.

First Report of Ciguatoxins in Two Starfish Species: Ophidiaster ophidianus and Marthasterias glacialis

PubMed Central

Silva, Marisa; Rodriguez, Inés; Barreiro, Aldo; Kaufmann, Manfred; Neto, Ana Isabel; Hassouani, Meryem; Sabour, Brahim; Alfonso, Amparo; Botana, Luis M.; Vasconcelos, Vitor

2015-01-01

Ciguatera fish poisoning (CFP) is a syndrome caused by the ingestion of fish contaminated with Ciguatoxins (CTXs). These phycotoxins are produced mainly by dinoflagellates that belong to the genus Gambierdiscus that are transformed in more toxic forms in predatory fish guts, and are more present in the Indo-Pacific and Caribbean areas. It is estimated that CFP causes per year more than 10,000 intoxications worldwide. With the rise of water temperature and anthropogenic intervention, it is important to study the prevalence of CFP in more temperate waters. Through inter- and subtidal sampling, 22 species of organisms were collected, in Madeira and Azores archipelagos and in the northwestern Moroccan coast, during September of 2012 and June and July of 2013. A total of 94 samples of 22 different species of bivalves, gastropods, echinoderms and crustaceans where analyzed by Ultra Performance Liquid Chromatography-Mass Spectometry-Ion Trap-Time of Flight (UPLC-MS-IT-TOF) and Ultra Performance Chromatography- Mass Spectrometry (UPLC-MS). Our main aim was to detect new vectors and ascertain if there were some geographical differences. We detected for the first time putative CTXs in echinoderms, in two starfish species—M. glacialis and O. ophidianus. We detected differences regarding uptake values by organisms and geographical location. Toxin amounts were significant, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, in order to better access human health risk. In addition, we suggest monitoring of these toxins should be extended to other vectors, starfish being a good alternative for protecting and accessing human health risk. PMID:26402702

Live-cell FRET imaging reveals clustering of the prion protein at the cell surface induced by infectious prions.

PubMed

Tavares, Evandro; Macedo, Joana A; Paulo, Pedro M R; Tavares, Catarina; Lopes, Carlos; Melo, Eduardo P

2014-07-01

Prion diseases are associated to the conversion of the prion protein into a misfolded pathological isoform. The mechanism of propagation of protein misfolding by protein templating remains largely unknown. Neuroblastoma cells were transfected with constructs of the prion protein fused to both CFP-GPI-anchored and to YFP-GPI-anchored and directed to its cell membrane location. Live-cell FRET imaging between the prion protein fused to CFP or YFP was measured giving consistent values of 10±2%. This result was confirmed by fluorescence lifetime imaging microscopy and indicates intermolecular interactions between neighbor prion proteins. In particular, considering that a maximum FRET efficiency of 17±2% was determined from a positive control consisting of a fusion CFP-YFP-GPI-anchored. A stable cell clone expressing the two fusions containing the prion protein was also selected to minimize cell-to-cell variability. In both, stable and transiently transfected cells, the FRET efficiency consistently increased in the presence of infectious prions - from 4±1% to 7±1% in the stable clone and from 10±2% to 16±1% in transiently transfected cells. These results clearly reflect an increased clustering of the prion protein on the membrane in the presence of infectious prions, which was not observed in negative control using constructs without the prion protein and upon addition of non-infected brain. Our data corroborates the recent view that the primary site for prion conversion is the cell membrane. Since our fluorescent cell clone is not susceptible to propagate infectivity, we hypothesize that the initial event of prion infectivity might be the clustering of the GPI-anchored prion protein. Copyright © 2014 Elsevier B.V. All rights reserved.

Contribution to the risk characterization of ciguatoxins: LOAEL estimated from eight ciguatera fish poisoning events in Guadeloupe (French West Indies).

PubMed

Hossen, Virginie; Soliño, Lucia; Leroy, Patricia; David, Eric; Velge, Pierre; Dragacci, Sylviane; Krys, Sophie; Flores Quintana, Harold; Diogène, Jorge

2015-11-01

From 2010 to 2012, 35 ciguatera fish poisoning (CFP) events involving 87 individuals who consumed locally-caught fish were reported in Guadeloupe (French West Indies). For 12 of these events, the presence of ciguatoxins (CTXs) was indicated in meal remnants and in uncooked fish by the mouse bioassay (MBA). Caribbean ciguatoxins (C-CTXs) were confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Using a cell-based assay (CBA), and the only available standard Pacific ciguatoxin-1 (P-CTX-1), the lowest toxins level detected in fish samples causing CFP was 0.022 µg P-CTX-1 equivalent (eq.)·kg(-1) fish. Epidemiological and consumption data were compiled for most of the individuals afflicted, and complete data for establishing the lowest observable adverse effects level (LOAEL) were obtained from 8 CFP events involving 21 individuals. Based on toxin intakes, the LOAEL was estimated at 4.2 ng P-CTX-1 eq./individual corresponding to 48. 4 pg P-CTX-1 eq.kg(-1) body weight (bw). Although based on limited data, these results are consistent with the conclusions of the European Food Safety Authority (EFSA) opinion which indicates that a level of 0.01 µg P-CTX-1 eq.kg(-1) fish, regardless of source, should not exert effects in sensitive individuals when consuming a single meal. The calculated LOAEL is also consistent with the U.S. Food and Drug Administration guidance levels for CTXs (0.1 µg C-CTX-1 eq.kg(-1) and 0.01 µg P-CTX-1 eq.kg(-1) fish). Copyright © 2015 Elsevier Inc. All rights reserved.

Transgenic carrot expressing fusion protein comprising M. tuberculosis antigens induces immune response in mice.

PubMed

Permyakova, Natalia V; Zagorskaya, Alla A; Belavin, Pavel A; Uvarova, Elena A; Nosareva, Olesya V; Nesterov, Andrey E; Novikovskaya, Anna A; Zav'yalov, Evgeniy L; Moshkin, Mikhail P; Deineko, Elena V

2015-01-01

Tuberculosis remains one of the major infectious diseases, which continues to pose a major global health problem. Transgenic plants may serve as bioreactors to produce heterologous proteins including antibodies, antigens, and hormones. In the present study, a genetic construct has been designed that comprises the Mycobacterium tuberculosis genes cfp10, esat6 and dIFN gene, which encode deltaferon, a recombinant analog of the human γ-interferon designed for expression in plant tissues. This construct was transferred to the carrot (Daucus carota L.) genome by Agrobacterium-mediated transformation. This study demonstrates that the fusion protein CFP10-ESAT6-dIFN is synthesized in the transgenic carrot storage roots. The protein is able to induce both humoral and cell-mediated immune responses in laboratory animals (mice) when administered either orally or by injection. It should be emphasized that M. tuberculosis antigens contained in the fusion protein have no cytotoxic effect on peripheral blood mononuclear cells.

Integration of lyoplate based flow cytometry and computational analysis for standardized immunological biomarker discovery.

PubMed

Villanova, Federica; Di Meglio, Paola; Inokuma, Margaret; Aghaeepour, Nima; Perucha, Esperanza; Mollon, Jennifer; Nomura, Laurel; Hernandez-Fuentes, Maria; Cope, Andrew; Prevost, A Toby; Heck, Susanne; Maino, Vernon; Lord, Graham; Brinkman, Ryan R; Nestle, Frank O

2013-01-01

Discovery of novel immune biomarkers for monitoring of disease prognosis and response to therapy in immune-mediated inflammatory diseases is an important unmet clinical need. Here, we establish a novel framework for immunological biomarker discovery, comparing a conventional (liquid) flow cytometry platform (CFP) and a unique lyoplate-based flow cytometry platform (LFP) in combination with advanced computational data analysis. We demonstrate that LFP had higher sensitivity compared to CFP, with increased detection of cytokines (IFN-γ and IL-10) and activation markers (Foxp3 and CD25). Fluorescent intensity of cells stained with lyophilized antibodies was increased compared to cells stained with liquid antibodies. LFP, using a plate loader, allowed medium-throughput processing of samples with comparable intra- and inter-assay variability between platforms. Automated computational analysis identified novel immunophenotypes that were not detected with manual analysis. Our results establish a new flow cytometry platform for standardized and rapid immunological biomarker discovery with wide application to immune-mediated diseases.

Integration of Lyoplate Based Flow Cytometry and Computational Analysis for Standardized Immunological Biomarker Discovery

PubMed Central

Villanova, Federica; Di Meglio, Paola; Inokuma, Margaret; Aghaeepour, Nima; Perucha, Esperanza; Mollon, Jennifer; Nomura, Laurel; Hernandez-Fuentes, Maria; Cope, Andrew; Prevost, A. Toby; Heck, Susanne; Maino, Vernon; Lord, Graham; Brinkman, Ryan R.; Nestle, Frank O.

2013-01-01

Discovery of novel immune biomarkers for monitoring of disease prognosis and response to therapy in immune-mediated inflammatory diseases is an important unmet clinical need. Here, we establish a novel framework for immunological biomarker discovery, comparing a conventional (liquid) flow cytometry platform (CFP) and a unique lyoplate-based flow cytometry platform (LFP) in combination with advanced computational data analysis. We demonstrate that LFP had higher sensitivity compared to CFP, with increased detection of cytokines (IFN-γ and IL-10) and activation markers (Foxp3 and CD25). Fluorescent intensity of cells stained with lyophilized antibodies was increased compared to cells stained with liquid antibodies. LFP, using a plate loader, allowed medium-throughput processing of samples with comparable intra- and inter-assay variability between platforms. Automated computational analysis identified novel immunophenotypes that were not detected with manual analysis. Our results establish a new flow cytometry platform for standardized and rapid immunological biomarker discovery with wide application to immune-mediated diseases. PMID:23843942

Some factors associated with change in patient-centredness of student nurses during the Common Foundation Programme in Nursing.

PubMed

Rolfe, G

1994-10-01

This longitudinal study examines the changes in patient-centredness of a group of student nurses during their Common Foundation Programme in Nursing, and attempts to explore some of the social, psychological and educational factors which might be associated with such change. The Patient-centredness Multi-choice Questionnaire (PMQX) was administered to a sample of 267 student nurses on commencement and completion of their Common Foundation Programme in Nursing, and significant positive as well as negative changes in PMQX scores were found, although the mean scores for the sample were unchanged. A number of factors were found to be associated with high levels of patient-centredness on commencement of the course, including marital status, age, and radicalism. Positive change in patient-centredness during the CFP was associated with a variety of variables such as class size, a student-centred climate and a tenderminded attitude. Other factors, such as psychological stability and extroversion, were associated with both increase and decrease in patient-centredness during the CFP.

Integrated Biorefining: Coproduction of Renewable Resol Biopolymer for Aqueous Stream Valorization

DOE PAGES

Wilson, A. Nolan; Price, Mariel J.; Mukarakate, Calvin; ...

2017-07-13

Phenol-formaldehyde resins are major material classes that are used in a range of applications including composites, adhesives, foams, electronics, and insulation. While efforts have been made to produce renewable resins, there has yet to be an approach that offers potential for economic viability and meets all critical quality metrics. This failure can be attributed largely to the use of phenol and cresol homologues and to high separation costs. In this work, the use of phenol, cresol, and alkyl phenols derived from the aqueous phase generated from catalytic fast pyrolysis of biomass to produce a high-quality biobased resin is demonstrated. Production,more » through catalytic fast pyrolysis (CFP), separation, through distillation and adsorption unit operations, and synthesis, through typical resol chemistry, produced a resin with properties, such as curing kinetics and molecular weight, competitive with petroleum-derived resin. In conclusion, this work explores a pathway to value-added coproducts from a CFP waste stream, which has the potential to improve the economic viability of biofuels production.« less

Integrated Biorefining: Coproduction of Renewable Resol Biopolymer for Aqueous Stream Valorization

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

Wilson, A. Nolan; Price, Mariel J.; Mukarakate, Calvin

Phenol-formaldehyde resins are major material classes that are used in a range of applications including composites, adhesives, foams, electronics, and insulation. While efforts have been made to produce renewable resins, there has yet to be an approach that offers potential for economic viability and meets all critical quality metrics. This failure can be attributed largely to the use of phenol and cresol homologues and to high separation costs. In this work, the use of phenol, cresol, and alkyl phenols derived from the aqueous phase generated from catalytic fast pyrolysis of biomass to produce a high-quality biobased resin is demonstrated. Production,more » through catalytic fast pyrolysis (CFP), separation, through distillation and adsorption unit operations, and synthesis, through typical resol chemistry, produced a resin with properties, such as curing kinetics and molecular weight, competitive with petroleum-derived resin. In conclusion, this work explores a pathway to value-added coproducts from a CFP waste stream, which has the potential to improve the economic viability of biofuels production.« less

Exploring dynamics of molybdate in living animal cells by a genetically encoded FRET nanosensor.

PubMed

Nakanishi, Yoichi; Iida, Syuntaro; Ueoka-Nakanishi, Hanayo; Niimi, Tomoaki; Tomioka, Rie; Maeshima, Masayoshi

2013-01-01

Molybdenum (Mo) is an essential trace element for almost all living organisms including animals. Mo is used as a catalytic center of molybdo-enzymes for oxidation/reduction reactions of carbon, nitrogen, and sulfur metabolism. Whilst living cells are known to import inorganic molybdate oxyanion from the surrounding environment, the in vivo dynamics of cytosolic molybdate remain poorly understood as no appropriate indicator is available for this trace anion. We here describe a genetically encoded Förester-resonance-energy-transfer (FRET)-based nanosensor composed of CFP, YFP and the bacterial molybdate-sensor protein ModE. The nanosensor MolyProbe containing an optimized peptide-linker responded to nanomolar-range molybdate selectively, and increased YFP:CFP fluorescence intensity ratio by up to 109%. By introduction of the nanosensor, we have been able to successfully demonstrate the real-time dynamics of molybdate in living animal cells. Furthermore, time course analyses of the dynamics suggest that novel oxalate-sensitive- and sulfate-resistant- transporter(s) uptake molybdate in a model culture cell.

Cyclo(valine-valine) inhibits Vibrio cholerae virulence gene expression.

PubMed

Vikram, Amit; Ante, Vanessa M; Bina, X Renee; Zhu, Qin; Liu, Xinyu; Bina, James E

2014-06-01

Vibrio cholerae has been shown to produce a cyclic dipeptide, cyclo(phenylalanine-proline) (cFP), that functions to repress virulence factor production. The objective of this study was to determine if heterologous cyclic dipeptides could repress V. cholerae virulence factor production. To that end, three synthetic cyclic dipeptides that differed in their side chains from cFP were assayed for virulence inhibitory activity in V. cholerae. The results revealed that cyclo(valine-valine) (cVV) inhibited virulence factor production by a ToxR-dependent process that resulted in the repression of the virulence regulator aphA. cVV-dependent repression of aphA was found to be independent of known aphA regulatory genes. The results demonstrated that V. cholerae was able to respond to exogenous cyclic dipeptides and implicated the hydrophobic amino acid side chains on both arms of the cyclo dipeptide scaffold as structural requirements for inhibitory activity. The results further suggest that cyclic dipeptides have potential as therapeutics for cholera treatment. © 2014 The Authors.

12 CFR 741.12 - Liquidity and contingency funding plans.

Code of Federal Regulations, 2014 CFR

2014-01-01

.../liability policy, a funds management policy, or a business continuity policy. The CFP must address, at a...; (3) Policies to manage a range of stress environments, identification of some possible stress events... the institution to respond to liquidity events; (5) Management processes that include clear...

Critical role of NMDA but not opioid receptors in the acquisition of fat-conditioned flavor preferences in rats.

PubMed

Dela Cruz, J A D; Bae, V S; Icaza-Cukali, D; Sampson, C; Bamshad, D; Samra, A; Singh, S; Khalifa, N; Touzani, K; Sclafani, A; Bodnar, R J

2012-11-01

Animals learn to prefer flavors associated with the intake of dietary fats such as corn oil (CO) solutions. We previously reported that fat-conditioned flavor preferences in rats were relatively unaffected by systemic treatment with dopamine D1 and D2 antagonsits. The present study examined whether systemic opioid (naltrexone, NTX) or NMDA (MK-801) receptor antagonists altered the acquisition and/or expression of CO-CFP. The CFP was produced by training rats to drink one novel flavor (CS+, e.g., cherry) mixed in a 3.5% CO solution and another flavor (CS-, e.g., grape) in a 0.9% CO solution. In expression studies, food-restricted rats drank these solutions in one-bottle sessions (2 h) over 10 d. Subsequent two-bottle tests with the CS+ and CS- flavors mixed in 0.9% CO solutions occurred 0.5h after systemic administration of vehicle (VEH), NTX (0.1-5 mg/kg) or MK-801 (50-200 μg/kg). Rats displayed a robust CS+ preference following VEH treatment (85-88%) which was significantly though moderately attenuated by NTX (69-70%). The lower doses of MK-801 slightly reduced the CS+ preference; the high dose blocked the CS+ preference (49%) but also markedly reduced overall CS intake. In separate acquisition studies, rats received VEH or NTX (0.1, 0.5, 1mg/kg) or MK-801 (100 μg/kg) 0.5h prior to 1-bottle training trials with CS+/3.5% CO and CS-/0.9% CO training solutions. Additional Limited VEH groups were trained with intakes limited to that of the NTX and MK-801 groups. Subsequent two-bottle CS+ vs. CS- tests were conducted without injections. Significant and persistent CS+ preferences were observed in VEH (77-84%) and Limited VEH (88%) groups. NTX treatment during training failed to block the acquisition of CO-CFP although the magnitude of the CS+ preference was reduced by 0.5 (70%) and 1.0 (72%) mg/kg doses relative to the Limited VEH treatment (88%). In contrast, MK-801 (100 μg/kg) treatment during training blocked the acquisition of the CO-CFP. These data suggest a critical role for NMDA, but not opioid receptor signaling in the acquisition of a fat conditioned flavor preferences, and at best limited involvement of NMDA and opioid receptors in the expression of a previously learned preference. Copyright © 2012 Elsevier Inc. All rights reserved.

Risk factors for increased immune reconstitution in response to Mycobacterium tuberculosis antigens in tuberculosis HIV-infected, antiretroviral-naïve patients.

PubMed

da Silva, Tatiana Pereira; Giacoia-Gripp, Carmem Beatriz Wagner; Schmaltz, Carolina A; Sant'Anna, Flavia Marinho; Saad, Maria Helena; Matos, Juliana Arruda de; de Lima E Silva, Julio Castro Alves; Rolla, Valeria Cavalcanti; Morgado, Mariza Gonçalves

2017-09-06

Little is known regarding the restoration of the specific immune response after combined antiretroviral therapy (cART) and anti-tuberculosis (TB) therapy introduction among TB-HIV patients. In this study, we examined the immune response of TB-HIV patients to Mycobacterium tuberculosis (Mtb) antigens to evaluate the response dynamics to different antigens over time. Moreover, we also evaluated the influence of two different doses of efavirenz and the factors associated with immune reconstitution. This is a longitudinal study nested in a clinical trial, where cART was initiated during the baseline visit (D0), which occurred 30 ± 10 days after the introduction of anti-TB therapy. Follow-up visits were performed at 30, 60, 90 and 180 days after cART initiation. The production of IFN-γ upon in vitro stimulation with Mtb antigens purified protein derivative (PPD), ESAT-6 and 38 kDa/CFP-10 using ELISpot was examined at baseline and follow-up visits. Sixty-one patients, all ART-naïve, were selected and included in the immune reconstitution analysis; seven (11.5%) developed Immune Reconstitution Inflammatory Syndrome (IRIS). The Mtb specific immune response was higher for the PPD antigen followed by 38 kDa/CFP-10 and increased in the first 60 days after cART initiation. In multivariate analysis, the variables independently associated with increased IFN-γ production in response to PPD antigen were CD4 + T cell counts <200 cells/mm 3 at baseline, age, site of tuberculosis, 800 mg efavirenz dose and follow-up CD4 + T cell counts. Moreover, the factors associated with the production of IFN-γ in response to 38 kDa/CFP-10 were detectable HIV viral load (VL) and CD4 + T cell counts at follow-up visits of ≥200 cells/mm 3 . These findings highlight the differences in immune response according to the specificity of the Mtb antigen, which contributes to a better understanding of TB-HIV immunopathogenesis. IFN-γ production elicited by PPD and 38 kDa/CFP-10 antigens have a greater magnitude compared to ESAT-6 and are associated with different factors. The low response to ESAT-6, even during immune restoration, suggests that this antigen is not adequate to assess the immune response of immunosuppressed TB-HIV patients.

A Laboratory Exercise for Visible Gel Filtration Chromatography Using Fluorescent Proteins

ERIC Educational Resources Information Center

Zhang, Wenqiang; Cao, Yibin; Xu, Lishan; Gong, Jufang; Sun, Meihao

2015-01-01

Gel filtration chromatography (GFC) separates molecules according to size and is one of the most widely used methods for protein purification. Here, red fluorescent protein (RFP), green fluorescent protein (GFP), yellow fluorescent protein (YFP), cyan fluorescent protein (CFP), and/or their fusion proteins were prokaryotically expressed, purified,…

36 CFR 230.6 - Project costs and cost share requirements.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 36 Parks, Forests, and Public Property 2 2013-07-01 2013-07-01 false Project costs and cost share... Project costs and cost share requirements. (a) The CFP Federal contribution cannot exceed 50 percent of the total project costs. (b) Allowable project and cost share costs will include the purchase price...

36 CFR 230.6 - Project costs and cost share requirements.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 36 Parks, Forests, and Public Property 2 2014-07-01 2014-07-01 false Project costs and cost share... Project costs and cost share requirements. (a) The CFP Federal contribution cannot exceed 50 percent of the total project costs. (b) Allowable project and cost share costs will include the purchase price...

36 CFR 230.6 - Project costs and cost share requirements.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 36 Parks, Forests, and Public Property 2 2012-07-01 2012-07-01 false Project costs and cost share... Project costs and cost share requirements. (a) The CFP Federal contribution cannot exceed 50 percent of the total project costs. (b) Allowable project and cost share costs will include the purchase price...

78 FR 23437 - Funding and Fiscal Affairs, Loan Policies and Operations, and Funding Operations; Liquidity and...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-18

...'s liquidity needs beyond 90 days; and Strengthen each bank's Contingency Funding Plan (CFP). II... more focused policies concerning liquidity management and the contingency funding plan; (3) Divide the... comprehensive revisions. In this context, all commenters expressed the view that the proposed rule is excessive...

Hybrid composite Ni(OH)2@NiCo2O4 grown on carbon fiber paper for high-performance supercapacitors.

PubMed

Huang, Liang; Chen, Dongchang; Ding, Yong; Wang, Zhong Lin; Zeng, Zhengzhi; Liu, Meilin

2013-11-13

We have successfully fabricated and tested the electrochemical performance of supercapacitor electrodes consisting of Ni(OH)2 nanosheets coated on NiCo2O4 nanosheets grown on carbon fiber paper (CFP) current collectors. When the NiCo2O4 nanosheets are replaced by Co3O4 nanosheets, however, the energy and power density as well as the rate capability of the electrodes are significantly reduced, most likely due to the lower conductivity of Co3O4 than that of NiCo2O4. The 3D hybrid composite Ni(OH)2/NiCo2O4/CFP electrodes demonstrate a high areal capacitance of 5.2 F/cm(2) at a cycling current density of 2 mA/cm(2), with a capacitance retention of 79% as the cycling current density was increased from 2 to 50 mA/cm(2). The remarkable performance of these hybrid composite electrodes implies that supercapacitors based on them have potential for many practical applications.

Crude Oil Remote Sensing, Characterization and Cleaning with CW and Pulsed Lasers

NASA Technical Reports Server (NTRS)

Kukhtareva, Tatiana; Chirita, Arc; Gallegos, Sonia C.

2014-01-01

For detection, identification and characterization of crude oil we combine several optical methods of remote sensing of crude oil films and emulsions (coherent fringe projection illumination (CFP), holographic in-line interferometry (HILI), and laser induced fluorescence). These methods allow the three-dimensional characterization of oil spills, important for practical applications. Combined methods of CFP and HILI are described in the frame of coherent superposition of partial interference patterns. It is shown, that in addition to detection/identification laser illumination in the green-blue region can also degrade oil slicks. Different types of surfaces contaminated by oil spills are tested: oil on the water, oil on the flat solid surfaces and oil on the curved surfaces of pipes. For the detection and monitoring of the laser-induced oil degradation in pipes, coherent fiber bundles were used. Both continuous-wave (CW) and pulsed lasers are tested using pump-probe schemes. This finding suggests that properly structured laser clean-up can be an alternative environmentally-friendly method of decontamination, as compared to the currently used chemical methods that are dangerous to environment.

Glycogen synthase kinase-3β facilitates cell apoptosis induced by high fluence low-power laser irradiation through acceleration of Bax translocation

NASA Astrophysics Data System (ADS)

Huang, Lei; Wu, Shengnan; Xing, Da

2011-03-01

Glycogen synthase kinase-3β (GSK-3β) is a critical activator of cell apoptosis induced by a diverse array of insults. However, the effects of GSK-3β on the human lung adenocarcinoma cell (ASTC-a-1) apoptosis induced by high fluence low-power laser irradiation (HF-LPLI) are not clear. Here, we showed that GSK-3β was constantly translocated from cytoplasm to nucleus and activated during HF-LPLI-induced cell apoptosis. In addition, we found that co-overexpression of YFP-GSK-3β and CFP-Bax in ASTC-a-1 cells accelerated both Bax translocations to mitochondria and cell apoptosis, compared to the cells expressed CFP-Bax only under HF-LPLI treatment, indicating that GSK-3β facilitated ASTC-a-1 cells apoptosis through acceleration mitochondrial translocation of Bax. Our results demonstrate that GSK-3β exerts some of its pro-apoptotic effects in ASTC-a-1 cells by regulating the mitochondrial localization of Bax, a key component of the intrinsic apoptotic cascade.

Direct multiplex imaging and optogenetics of Rho GTPases enabled by near-infrared FRET.

PubMed

Shcherbakova, Daria M; Cox Cammer, Natasha; Huisman, Tsipora M; Verkhusha, Vladislav V; Hodgson, Louis

2018-06-01

Direct visualization and light control of several cellular processes is a challenge, owing to the spectral overlap of available genetically encoded probes. Here we report the most red-shifted monomeric near-infrared (NIR) fluorescent protein, miRFP720, and the fully NIR Förster resonance energy transfer (FRET) pair miRFP670-miRFP720, which together enabled design of biosensors compatible with CFP-YFP imaging and blue-green optogenetic tools. We developed a NIR biosensor for Rac1 GTPase and demonstrated its use in multiplexed imaging and light control of Rho GTPase signaling pathways. Specifically, we combined the Rac1 biosensor with CFP-YFP FRET biosensors for RhoA and for Rac1-GDI binding, and concurrently used the LOV-TRAP tool for upstream Rac1 activation. We directly observed and quantified antagonism between RhoA and Rac1 dependent on the RhoA-downstream effector ROCK; showed that Rac1 activity and GDI binding closely depend on the spatiotemporal coordination between these two molecules; and simultaneously observed Rac1 activity during optogenetic manipulation of Rac1.

Early detection of the growth of Mycobacterium tuberculosis using magnetophoretic immunoassay in liquid culture.

PubMed

Kim, Jeonghyo; Lee, Kil-Soo; Kim, Eun Bee; Paik, Seungwha; Chang, Chulhun L; Park, Tae Jung; Kim, Hwa-Jung; Lee, Jaebeom

2017-10-15

Tuberculosis (TB) is an often neglected, epidemic disease that remains to be controlled by contemporary techniques of medicine and biotechnology. In this study, a nanoscale sensing system, referred to as magnetophoretic immunoassay (MPI) was designed to capture culture filtrate protein (CFP)-10 antigens effectively using two different types of nanoparticles (NPs). Two specific monoclonal antibodies against CFP-10 antigen were used, including gold NPs for signaling and magnetic particles for separation. These results were carefully compared with those obtained using the commercial mycobacteria growth indicator tube (MGIT) test via 2 sequential clinical tests (with ca. 260 clinical samples). The sensing linearity of MPI was shown in the range of pico- to micromoles and the detection limit was 0.3pM. MPI using clinical samples shows robust and reliable sensing while monitoring Mycobacterium tuberculosis (MTB) growth with monitoring time 3-10 days) comparable to that with the MGIT test. Furthermore, MPI distinguished false-positive samples from MGIT-positive samples, probably containing non-tuberculous mycobacteria. Thus, MPI shows promise in early TB diagnosis. Copyright © 2017 Elsevier B.V. All rights reserved.

Probing Protein Structure in Vivo with FRET

PubMed Central

Davis, Trisha; Muller, Eric

2012-01-01

Fluorescence resonance energy transfer (FRET) is widely used to construct probes for cellular activities and to complement two-hybrid results that predict protein-protein interactions. The Yeast Resource Center promotes an underutilized potential of FRET as an in vivo tool to position proteins within low resolution structures derived from electron microscopy. The success of this approach using widefield microscopy depends upon the choice of filter sets, standardized image acquisition, a robust metric and controls matched to the structure under investigation. A comparison of various CFP and YFP filter combinations from Chroma and Semrock demonstrated the strength of the Chroma filters when coupled with our FRET metric, termed FretR. Coupling CFP and YFP to a selection of proteins of known structure allowed us to create a standard curve of FretR versus distance. How well other FRET metrics conform was also evaluated. Finally FretR was linked to an approximation of the efficiency of energy transfer. Together this feature set has allowed us to contribute to our understanding of the organization of the yeast spindle pole body, cohesin complex and gamma-tubulin complex.

Practical three color live cell imaging by widefield microscopy

PubMed Central

Xia, Jianrun; Kim, Song Hon H.; Macmillan, Susan

2006-01-01

Live cell fluorescence microscopy using fluorescent protein tags derived from jellyfish and coral species has been a successful tool to image proteins and dynamics in many species. Multi-colored aequorea fluorescent protein (AFP) derivatives allow investigators to observe multiple proteins simultaneously, but overlapping spectral properties sometimes require the use of sophisticated and expensive microscopes. Here, we show that the aequorea coerulescens fluorescent protein derivative, PS-CFP2 has excellent practical properties as a blue fluorophore that are distinct from green or red fluorescent proteins and can be imaged with standard filter sets on a widefield microscope. We also find that by widefield illumination in live cells, that PS-CFP2 is very photostable. When fused to proteins that form concentrated puncta in either the cytoplasm or nucleus, PSCFP2 fusions do not artifactually interact with other AFP fusion proteins, even at very high levels of over-expression. PSCFP2 is therefore a good blue fluorophore for distinct three color imaging along with eGFP and mRFP using a relatively simple and inexpensive microscope. PMID:16909160

RNA aptamers that functionally interact with green fluorescent protein and its derivatives

PubMed Central

Shui, Bo; Ozer, Abdullah; Zipfel, Warren; Sahu, Nevedita; Singh, Avtar; Lis, John T.; Shi, Hua; Kotlikoff, Michael I.

2012-01-01

Green Fluorescent Protein (GFP) and related fluorescent proteins (FPs) have been widely used to tag proteins, allowing their expression and subcellular localization to be examined in real time in living cells and animals. Similar fluorescent methods are highly desirable to detect and track RNA and other biological molecules in living cells. For this purpose, we have developed a group of RNA aptamers that bind GFP and related proteins, which we term Fluorescent Protein-Binding Aptamers (FPBA). These aptamers bind GFP, YFP and CFP with low nanomolar affinity and binding decreases GFP fluorescence, whereas slightly augmenting YFP and CFP brightness. Aptamer binding results in an increase in the pKa of EGFP, decreasing the 475 nm excited green fluorescence at a given pH. We report the secondary structure of FPBA and the ability to synthesize functional multivalent dendrimers. FPBA expressed in live cells decreased GFP fluorescence in a valency-dependent manner, indicating that the RNA aptamers function within cells. The development of aptamers that bind fluorescent proteins with high affinity and alter their function, markedly expands their use in the study of biological pathways. PMID:22189104

Hybrid Composite Ni(OH)(2)@NiCo2O4 Grown on Carbon Fiber Paper for High-Performance Supercapacitors

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

Huang, L; Chen, DC; Ding, Y

2013-11-13

We have successfully fabricated and tested the electrochemical performance of supercapacitor electrodes consisting of Ni(OH)(2) nanosheets coated on NiCo2O4 nanosheets grown on carbon fiber paper (CFP) current collectors. When the NiCo2O4 nanosheets are replaced by Co3O4 nanosheets, however, the energy and power density as well as the rate capability of the electrodes are significantly reduced, most likely due to the lower conductivity of Co3O4 than that of NiCo2O4. The 3D hybrid composite Ni(OH)(2)/ NiCo2O4/CFP electrodes demonstrate a high areal capacitance of 5.2 F/cm(2) at a cycling current density of 2 rnA/cm(2), with a capacitance retention of 79% as the cyclingmore » current density was increased from 2 to 50 mA/cm(2). The remarkable performance of these hybrid composite electrodes implies that supercapacitors based on them have potential for many practical applications.« less

Experimental and computational investigation of acetic acid deoxygenation over oxophilic molybdenum carbide: Surface chemistry and active site identity

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

Schaidle, Joshua A.; Blackburn, Jeffrey; Farberow, Carrie A.

Ex situ catalytic fast pyrolysis (CFP) is a promising route for producing fungible biofuels; however, this process requires bifunctional catalysts that favor C–O bond cleavage, activate hydrogen at near atmospheric pressure and high temperature (350–500 °C), and are stable under high-steam, low hydrogen-to-carbon environments. Recently, early transition-metal carbides have been reported to selectively cleave C–O bonds of alcohols, aldehydes, and oxygenated aromatics, yet there is limited understanding of the metal carbide surface chemistry under reaction conditions and the identity of the active sites for deoxygenation. In this study, we evaluated molybdenum carbide (Mo 2C) for the deoxygenation of acetic acid,more » an abundant component of biomass pyrolysis vapors, under ex situ CFP conditions, and we probed the Mo 2C surface chemistry, identity of the active sites, and deoxygenation pathways using in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), X-ray photoelectron spectroscopy (XPS), and density functional theory (DFT) calculations.« less

Hyperspectral imaging for simultaneous measurements of two FRET biosensors in pancreatic β-cells.

PubMed

Elliott, Amicia D; Bedard, Noah; Ustione, Alessandro; Baird, Michelle A; Davidson, Michael W; Tkaczyk, Tomasz; Piston, David W

2017-01-01

Fluorescent protein (FP) biosensors based on Förster resonance energy transfer (FRET) are commonly used to study molecular processes in living cells. There are FP-FRET biosensors for many cellular molecules, but it remains difficult to perform simultaneous measurements of multiple biosensors. The overlapping emission spectra of the commonly used FPs, including CFP/YFP and GFP/RFP make dual FRET measurements challenging. In addition, a snapshot imaging modality is required for simultaneous imaging. The Image Mapping Spectrometer (IMS) is a snapshot hyperspectral imaging system that collects high resolution spectral data and can be used to overcome these challenges. We have previously demonstrated the IMS's capabilities for simultaneously imaging GFP and CFP/YFP-based biosensors in pancreatic β-cells. Here, we demonstrate a further capability of the IMS to image simultaneously two FRET biosensors with a single excitation band, one for cAMP and the other for Caspase-3. We use these measurements to measure simultaneously cAMP signaling and Caspase-3 activation in pancreatic β-cells during oxidative stress and hyperglycemia, which are essential components in the pathology of diabetes.

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

Fu Guo; Institute of Neuroscience, Department of Neurobiology, Second Military Medical University, Shanghai 200433; Yang Huayan

Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to {beta}{sub 2} subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasmamore » membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we suggested that the conformation of Mac-1 subunits is not affected by the fusion of CFP or YFP and inferred that Mac-1 subunits take different conformation when expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293T cells, respectively.« less

Experimental and computational investigation of acetic acid deoxygenation over oxophilic molybdenum carbide: Surface chemistry and active site identity

DOE PAGES

Schaidle, Joshua A.; Blackburn, Jeffrey; Farberow, Carrie A.; ...

2016-01-21

Ex situ catalytic fast pyrolysis (CFP) is a promising route for producing fungible biofuels; however, this process requires bifunctional catalysts that favor C–O bond cleavage, activate hydrogen at near atmospheric pressure and high temperature (350–500 °C), and are stable under high-steam, low hydrogen-to-carbon environments. Recently, early transition-metal carbides have been reported to selectively cleave C–O bonds of alcohols, aldehydes, and oxygenated aromatics, yet there is limited understanding of the metal carbide surface chemistry under reaction conditions and the identity of the active sites for deoxygenation. In this study, we evaluated molybdenum carbide (Mo 2C) for the deoxygenation of acetic acid,more » an abundant component of biomass pyrolysis vapors, under ex situ CFP conditions, and we probed the Mo 2C surface chemistry, identity of the active sites, and deoxygenation pathways using in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), X-ray photoelectron spectroscopy (XPS), and density functional theory (DFT) calculations.« less

Induction of AID-targeting adaptor 14-3-3γ is mediated by NF-κB-dependent recruitment of CFP1 to the 5′-CpG-3′-rich 14-3-3γ promoter and is sustained by E2A

PubMed Central

Mai, Thach; Pone, Egest J.; Li, Guideng; Lam, Tonika S.; Moehlman, J’aime; Xu, Zhenming; Casali, Paolo

2013-01-01

Class switch DNA recombination (CSR) crucially diversifies antibody biological effectors functions. 14-3-3γ specifically binds to the 5′-AGCT-3′ repeats in the IgH locus switch (S) regions. By directly interacting with the C-terminal region of activation-induced cytidine deaminase (AID), 14-3-3γ targets this enzyme to S regions to mediate CSR. Here, we showed that 14-3-3γ was expressed in germinal center B cells in vivo and induced in B cells by T-dependent and T-independent primary CSR-inducing stimuli in vitro in humans and mice. Induction of 14-3-3γ was rapid, peaking within 3 h of stimulation by lipopolysaccharides (LPS), and sustained over the course of AID and CSR induction. It was dependent on recruitment of NF-κB to the 14-3-3γ gene promoter. The NF-κB recruitment enhanced the occupancy of the CpG island within the 14-3-3γ promoter by CFP1, a component of the COMPASS histone methyltransferase complex, and promoter-specific enrichment of histone 3 lysine 4 trimethylation (H3K4me3), which is indicative of open chromatin state and marks transcription-competent promoters. NF-κB also potentiated the binding of B cell lineage-specific factor E2A to an E-box motif located immediately downstream of the two closely-spaced transcription start sites (TSSs) for sustained 14-3-3γ expression and CSR induction. Thus, 14-3-3γ induction in CSR is enabled by the CFP1-mediated H3K4me3 enrichment in the promoter, dependent on NF-κB and sustained by E2A. PMID:23851690

Filarial infection modulates the immune response to Mycobacterium tuberculosis through expansion of CD4+ IL-4 memory T cells

PubMed Central

Chatterjee, Soumya; Clark, Carolyn E.; Lugli, Enrico; Roederer, Mario; Nutman, Thomas B.

2015-01-01

Exaggerated CD4+T helper 2-specific cytokine producing memory T cell responses developing concomitantly with a T helper1 response might have a detrimental role in immunity to infection caused by Mycobacterium tuberculosis (Mtb). To assess the dynamics of antigen (Ag)-specific memory T cell compartments in the context of filarial infection we used multiparameter flow cytometry on PBMCs from 25 microfilaremic filarial -infected (Inf) and 14 filarial-uninfected (Uninf) subjects following stimulation with filarial (BmA) or with the Mycobacterium tuberculosis (Mtb)-specific Ag CFP10. Our data demonstrated that the Inf group not only had a marked increase in BmA-specific CD4+IL-4+ cells (Median net frequency compared to baseline (Fo)=0.09% vs. 0.01%, p=0.038) but also to CFP10 (Fo =0.16% vs. 0.007%, p=0.04) and Staphylococcal Enterotoxin B (SEB) (Fo =0.49% vs. 0.26%, p=0.04). The Inf subjects showed a BmA-specific expansion of CD4+CD45RO+IL-4+ producing central memory (TCM, CD45RO+CCR7+CD27+) (Fo =1.1% vs. 0.5%, p=0.04) as well as effector memory (TEM CD45RO+CCR7-CD27-) (Fo =1.5% vs. 0.2%, p=0.03) with a similar but non-significant response to CFP10. In addition, there was expansion of CD4+ IL-4+ CD45RA+ CCR7+CD27+ (naïve-like) in Inf individuals compared to Uninf subjects. Among Inf subjects with definitive latent tuberculosis , there were no differences in frequencies of IL-4 producing cells within any of the memory compartments compared to the Uninf group. Our data suggest that filarial infection induces antigen-specific, exaggerated IL-4 responses in distinct T cell memory compartments to Mtb-specific antigens, which are attenuated in subjects who are able to mount a delayed type hypersensitivity reaction to Mtb. PMID:25667413

The 50-kDa protein of Apple chlorotic leaf spot virus interferes with intracellular and intercellular targeting and tubule-inducing activity of the 39-kDa protein of Grapevine berry inner necrosis virus.

PubMed

Isogai, M; Saitou, Y; Takahashi, N; Itabashi, T; Terada, M; Satoh, H; Yoshikawa, N

2003-03-01

To understand why transgenic Nicotiana occidentalis plants expressing a functional movement protein (MP) of Apple chlorotic leaf spot virus (ACLSV) show specific resistance to Grapevine berry inner necrosis virus (GINV), the MPs of ACLSV (50KP) and GINV (39KP) were fused to green, yellow, or cyan fluorescent proteins (GFP, YFP, or CFP). These fusion proteins were transiently expressed in leaf cells of both transgenic (50KP) and nontransgenic (NT) plants, and the intracellular and intercellular trafficking and tubule-inducing activity of these proteins were compared. The results indicate that in epidermal cells and protoplasts from 50KP plant leaves, the trafficking and tubule-inducing activities of GINV-39KP were specifically blocked while those of ACLSV-50KP and Apple stem grooving virus MP (36KP) were not affected. Additionally, when 39KP-YFP and 50KP-CFP were coexpressed in the leaf epidermis of NT plants, the fluorescence of both proteins was confined to single cells, indicating that 50KP-CFP interferes with the cell-to-cell trafficking of 39KP-YFP and vice versa. Mutational analyses of 50KP showed that the deletion mutants that retained the activities described above still blocked cell-to-cell trafficking of 39KP, but the dysfunctional 50KP mutants could no longer impede cell-to-cell movement of 39KP. Transgenic plants expressing the functional 50KP deletion mutants showed specific resistance against GINV. In contrast, transgenic plants expressing the dysfunctional 50KP mutants did not show any resistance to the virus. From these results, we conclude that the specific resistance of 50KP plants to GINV is due to the ability of the 50KP to block intracellular and intercellular trafficking of GINV 39KP.

Analysis of violet-excited fluorochromes by flow cytometry using a violet laser diode.

PubMed

Telford, William G; Hawley, Teresa S; Hawley, Robert G

2003-07-01

Low power violet laser diodes (VLDs) have been evaluated as potential replacements for water-cooled argon-ion and krypton-ion ultraviolet and violet lasers for DNA content analysis using the Hoechst dyes and 4,6-diamidino-2-phenylindole (Shapiro HMN, Perlmutter NG: Cytometry 44:133-136, 2001). In this study, we used a VLD to excite a variety of violet-excited fluorescent molecules important in biomedical analysis, including the fluorochromes Cascade Blue and Pacific Blue, the expressible fluorescent protein cyan fluorescent protein (CFP), and the fluorogenic alkaline phosphatase (AP) substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazoline (ELF-97; for endogenous AP detection and cell surface labeling with AP-conjugated antibodies). Comparisons were made between VLD excitation and a krypton-ion laser emitting at 407 nm (both at higher power levels and with the beam attenuated at levels approximating the VLD) on the same FACSVantage SE stream-in-air flow cytometer. We evaluated a Power Technology 408-nm VLD (30 mW) equipped with circularization optics (18 mW maximum output, set to 15 mW) and a Coherent I-302C krypton-ion laser emitting at power levels ranging from 15 to 75 mW. Cascade Blue, Pacific Blue, and CFP showed comparable signal-to-noise ratios and levels of sensitivity with VLD excitation versus the krypton-ion laser at high and VLD-matched power outputs. Multicolor fluorescent protein analysis with 488-nm excitation of green fluorescent protein and DsRed and VLD excitation of CFP was therefore feasible and was demonstrated. Similar levels of excitation efficiency between krypton-ion and VLD sources also were observed for ELF-97 detection. These evaluations confirmed that VLDs may be cost- and maintenance-effective replacements for water-cooled gas lasers for applications requiring violet excitation in addition to DNA binding dyes. Published 2003 Wiley-Liss, Inc.

Hierarchical 3-dimensional nickel-iron nanosheet arrays on carbon fiber paper as a novel electrode for non-enzymatic glucose sensing.

PubMed

Kannan, Palanisamy; Maiyalagan, Thandavarayan; Marsili, Enrico; Ghosh, Srabanti; Niedziolka-Jönsson, Joanna; Jönsson-Niedziolka, Martin

2016-01-14

Three-dimensional nickel-iron (3-D/Ni-Fe) nanostructures are exciting candidates for various applications because they produce more reaction-active sites than 1-D and 2-D nanostructured materials and exhibit attractive optical, electrical and catalytic properties. In this work, freestanding 3-D/Ni-Fe interconnected hierarchical nanosheets, hierarchical nanospheres, and porous nanospheres are directly grown on a flexible carbon fiber paper (CFP) substrate by a single-step hydrothermal process. Among the nanostructures, 3-D/Ni-Fe interconnected hierarchical nanosheets show excellent electrochemical properties because of its high conductivity, large specific active surface area, and mesopores on its walls (vide infra). The 3-D/Ni-Fe hierarchical nanosheet array modified CFP substrate is further explored as a novel electrode for electrochemical non-enzymatic glucose sensor application. The 3-D/Ni-Fe hierarchical nanosheet arrays exhibit significant catalytic activity towards the electrochemical oxidation of glucose, as compared to the 3-D/Ni-Fe hierarchical nanospheres, and porous nanospheres. The 3-D/Ni-Fe hierarchical nanosheet arrays can access a large amount of glucose molecules on their surface (mesopore walls) for an efficient electrocatalytic oxidation process. Moreover, 3-D/Ni-Fe hierarchical nanosheet arrays showed higher sensitivity (7.90 μA μM(-1) cm(-2)) with wide linear glucose concentration ranging from 0.05 μM to 0.2 mM, and the low detection limit (LOD) of 0.031 μM (S/N = 3) is achieved by the amperometry method. Further, the 3-D/Ni-Fe hierarchical nanosheet array modified CFP electrode can be demonstrated to have excellent selectivity towards the detection of glucose in the presence of 500-fold excess of major important interferents. All these results indicate that 3-D/Ni-Fe hierarchical nanosheet arrays are promising candidates for non-enzymatic glucose sensing.

Collaborative Aerospace Research and Fellowship Program at NASA Glenn Research Center

NASA Technical Reports Server (NTRS)

Heyward, Ann O.; Kankam, Mark D.

2004-01-01

During the summer of 2004, a 10-week activity for university faculty entitled the NASA-OAI Collaborative Aerospace Research and Fellowship Program (CFP) was conducted at the NASA Glenn Research Center in collaboration with the Ohio Aerospace Institute (OAI). This is a companion program to the highly successful NASA Faculty Fellowship Program and its predecessor, the NASA-ASEE Summer Faculty Fellowship Program that operated for 38 years at Glenn. The objectives of CFP parallel those of its companion, viz., (1) to further the professional knowledge of qualified engineering and science faculty,(2) to stimulate an exchange of ideas between teaching participants and employees of NASA, (3) to enrich and refresh the research and teaching activities of participants institutions, and (4) to contribute to the research objectives of Glenn. However, CFP, unlike the NASA program, permits faculty to be in residence for more than two summers and does not limit participation to United States citizens. Selected fellows spend 10 weeks at Glenn working on research problems in collaboration with NASA colleagues and participating in related activities of the NASA-ASEE program. This year's program began officially on June 1, 2004 and continued through August 7, 2004. Several fellows had program dates that differed from the official dates because university schedules vary and because some of the summer research projects warranted a time extension beyond the 10 weeks for satisfactory completion of the work. The stipend paid to the fellows was $1200 per week and a relocation allowance of $1000 was paid to those living outside a 50-mile radius of the Center. In post-program surveys from this and previous years, the faculty cited numerous instances where participation in the program has led to new courses, new research projects, new laboratory experiments, and grants from NASA to continue the work initiated during the summer. Many of the fellows mentioned amplifying material, both in undergraduate and graduate courses, on the basis of the summer s experience at Glenn. A number of 2004 fellows indicated that proposals to NASA will grow out of their summer research projects. In addition, some journal articles and NASA publications will result from this past summer s activities. Fellows from past summers continue to send reprints of articles that resulted from work initiated at Glenn. This report is intended primarily to summarize the research activities comprising the 2004 CFP Program at Glenn. Particular research studies include: 1) Development of an Imaging-Based, Computational Fluid Dynamics Tool to Assess Fluid Mechanics in Experimental Models that Simulate Blood Vessels; 2) Analysis of Nanomaterials Produced from Precursors; and 3) LEO Propagation Analysis Tool.

Catalytic and non-catalytic pyrolysis of biomass in non-inert environments for production of deoxygenated bio-oil and chemicals

USDA-ARS?s Scientific Manuscript database

Fast pyrolysis processes are among the most effective methods for liquefaction of lignocellulosic biomass. Catalytic fast pyrolysis (CFP) over HZSM-5 or other zeolites and/or utilization of reactive atmospheres such as in the non-catalytic Tail Gas Reactive Pyrolysis (TRGP) process, a recent patent...

24 CFR 905.120 - Penalties for slow obligation or expenditure of CFP assistance.

Code of Federal Regulations, 2010 CFR

2010-04-01

..., DEPARTMENT OF HOUSING AND URBAN DEVELOPMENT THE PUBLIC HOUSING CAPITAL FUND PROGRAM § 905.120 Penalties for... the PHA not later than September 30, 1999. (b) Exceptions to obligation requirement—(1) Extension... 12 months, based on: (i) The size of the PHA; (ii) The complexity of the capital program of the PHA...

Differential toxin profiles of ciguatoxins in marine organisms: Chemistry, fate and global distribution.

PubMed

Soliño, Lucía; Costa, Pedro Reis

2018-05-17

Ciguatoxins (CTXs) are fish metabolism products and a result of biotransformation of precursor gambiertoxins produced, in the first instance, by benthic dinoflagellates Gambierdiscus and Fukuyoa. Ciguatoxins are potent neurotoxins that selectively open voltage gated sodium channels in excitable cells causing the human food poisoning known as Ciguatera (CFP). Endemic from tropical areas in central Pacific and West Indian Ocean, and the Caribbean Sea, CTX may affect up to 500,000 people annually due to fish consumption. Their recent occurrence in European waters highlights the need for a multidisciplinary approach of CTX research in order to better understand the diversity and transformation of microalgae products through food webs. This article intends to review available information on chemistry, toxicity, distribution and fate of known CTX compounds from a critical perspective to provide an overview of future trends and needs on ciguatera research. Copyright © 2018 Elsevier Ltd. All rights reserved.

Harmful algal blooms and eutrophication: Examining linkages from selected coastal regions of the United States

PubMed Central

Anderson, Donald M.; Burkholder, JoAnn M.; Cochlan, William P.; Glibert, Patricia M.; Gobler, Christopher J.; Heil, Cynthia A.; Kudela, Raphael; Parsons, Michael L.; Rensel, J. E. Jack; Townsend, David W.; Trainer, Vera L.; Vargo, Gabriel A.

2008-01-01

Coastal waters of the United States (U.S.) are subject to many of the major harmful algal bloom (HAB) poisoning syndromes and impacts. These include paralytic shellfish poisoning (PSP), neurotoxic shellfish poisoning (NSP), amnesic shellfish poisoning (ASP), ciguatera fish poisoning (CFP) and various other HAB phenomena such as fish kills, loss of submerged vegetation, shellfish mortalities, and widespread marine mammal mortalities. Here, the occurrences of selected HABs in a selected set of regions are described in terms of their relationship to eutrophication, illustrating a range of responses. Evidence suggestive of changes in the frequency, extent or magnitude of HABs in these areas is explored in the context of the nutrient sources underlying those blooms, both natural and anthropogenic. In some regions of the U.S., the linkages between HABs and eutrophication are clear and well documented, whereas in others, information is limited, thereby highlighting important areas for further research. PMID:19956363

Effect of skin test on serum antibody responses to Mycobacterium bovis infection in cattle

USDA-ARS?s Scientific Manuscript database

Recently, several serologic tests designed to detect immunodominant antibodies to M. bovis antigens (e.g., MPB83, MPB70, ESAT-6, and CFP10) have emerged for potential use with samples from cattle. Of these, a commercial ELISA to MPB83/MPB70 (M. bovis antibody ELISA) has gained approval for use in ca...

Deactivation of Multilayered MFI Nanosheet Zeolite during Upgrading of Biomass Pyrolysis Vapors

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

Xu, Mengze; Mukarakate, Calvin; Iisa, Kristiina

Here, the catalytic fast pyrolysis (CFP) of biomass is a promising technology for producing renewable transportation fuels and chemicals. MFI-type catalysts have shown promise for CFP because they produce gasoline range hydrocarbons from oxygenated pyrolysis compounds; however, rapid catalyst deactivation due to coking is one of the major technical barriers inhibiting the commercialization of this technology. Coke deposited on the surface of the catalysts blocks access to active sites in the micropores leading to rapid catalyst deactivation. Our strategy is to minimize rapid catalyst deactivation by adding mesoporosity through forming MFI nanosheet materials. The synthesized MFI nanosheet catalysts were fullymore » characterized and evaluated for cellulose pyrolysis vapor upgrading to produce olefins and aromatic hydrocarbons. The data obtained from pyrolysis-GCMS (py-GCMS), showed that fresh MFI nanosheets produced similar aromatic hydrocarbon and olefin yields compared to conventional HZSM-5. However, MFI nanosheets demonstrated a longer lifetime than HZSM-5 even though coke contents were also higher than for HZSM-5 because the mesopores enabled better accessibility to active acid sites. This conclusion was supported by results from post-reaction analysis of various spent catalysts collected at different points during the deactivation experiments.« less

Architecture-dependent surface chemistry for Pt monolayers on carbon-supported Au.

PubMed

Cheng, Shuang; Rettew, Robert E; Sauerbrey, Marc; Alamgir, Faisal M

2011-10-01

Pt monolayers were grown by surface-limited redox replacement (SLRR) on two types of Au nanostructures. The Au nanostructures were fabricated electrochemically on carbon fiber paper (CFP) by either potentiostatic deposition (PSD) or potential square wave deposition (PSWD). The morphology of the Au/CFP heterostructures, examined using scanning electron microscopy (SEM), was found to depend on the type of Au growth method employed. The properties of the Pt deposit, as studied using X-ray photoelectron spectroscopy (XPS), X-ray absorption spectroscopy (XAS), and cyclic voltammetry (CV), were found to depend strongly on the morphology of the support. Specifically, it was found that smaller Au morphologies led to a higher degree of cationicity in the resulting Pt deposit, with Pt(4+) and Pt(2+) species being identified using XPS and XAS. For fuel-cell catalysts, the resistance of ultrathin catalyst deposits to surface area loss through dissolution, poisoning, and agglomeration is critical. This study shows that an equivalent of two monolayers (ML) is the low-loading limit of Pt on Au. At 1 ML or below, the Pt film decreases in activity and durability very rapidly due to presence of cationic Pt. © 2011 American Chemical Society

Observation of interaction between bid and 14-3-3 proteins by FRET in living cell during TNF-a-induced apoptosis

NASA Astrophysics Data System (ADS)

Wang, Jinjun; Chen, Tongsheng; Xing, Da; Wang, Fang

2005-01-01

Caspase8 is activated and cleaves Bid into two fragments when cells are exposed to death-inducing molecules such as tumor necrosis factor-α (TNF-α). Then the C-terminal fragment relocates from cytosol to mitochondria and promotes the release of cytochrome c, in the final cellular apoptosis is induced. Despite recent progress in the study of Bid during apoptosis induction, it remains unclear how C-terminal fragment of Bid cleaved moves to mitochondria and then induces the release of cytochrome c and so on. The 14-3-3 proteins are known to sequester certain pro-apoptotic members of Bcl-2 family. In order to further study the biological action of Bid during apoptosis, especially under physiological condition of living cell, the plasmids pBid-CFP and pYFP-14-3-3 were constructed. By the transient transfection of pBid-CFP and pYFP-14-3-3, the dynamic process of interaction of Bid and 14-3-3 protein in individual living cell during the apoptosis was primarily investigated with FRET (fluorescent resonance energy transfer) technique by the use of fluorescence microscopy.

Biosynthesis of Pd-Au alloys on carbon fiber paper: Towards an eco-friendly solution for catalysts fabrication

NASA Astrophysics Data System (ADS)

Zhuang, Zechao; Wang, Feifeng; Naidu, Ravendra; Chen, Zuliang

2015-09-01

Bimetallic nanomaterials with enhanced activity and stability have been extensively studied as emerging catalysts for hydrogen evolution reaction (HER). Expensive and environmentally unfriendly chemical synthesis routes inhibit their large-scale applications. In this work, we developed a facile and green synthesis of Pd-Au alloy nanoparticles (NPs) dispersed on carbon fiber paper (CFP) by plant-mediated bioreduction coupled with self-assembly. Engineering the morphology and composition of bimetallic catalysts synthesized by plant extracts on complex substrate is achieved. The resulting NPs are uniform in shape and have a spherical morphology with an average diameter of ∼180 nm, in which the molar ratio of Au/Pd is near 75:25 and the catalysts loading is about 0.5 mg cm-2. The Pd-Au/CFP hybrid electrode exhibits an excellent HER performance with a Tafel slope of 47 mV dec-1 and an exchange current density of 0.256 mA cm-2. Electrochemical stability tests through long-term potential cycles and potentiostatic electrolysis further confirm the high durability of the electrode. This development offers an efficient and eco-friendly catalysts synthesis route for constructing water-splitting cells and other electrocatalytic devices.

Deactivation of Multilayered MFI Nanosheet Zeolite during Upgrading of Biomass Pyrolysis Vapors

DOE PAGES

Xu, Mengze; Mukarakate, Calvin; Iisa, Kristiina; ...

2017-05-02

Here, the catalytic fast pyrolysis (CFP) of biomass is a promising technology for producing renewable transportation fuels and chemicals. MFI-type catalysts have shown promise for CFP because they produce gasoline range hydrocarbons from oxygenated pyrolysis compounds; however, rapid catalyst deactivation due to coking is one of the major technical barriers inhibiting the commercialization of this technology. Coke deposited on the surface of the catalysts blocks access to active sites in the micropores leading to rapid catalyst deactivation. Our strategy is to minimize rapid catalyst deactivation by adding mesoporosity through forming MFI nanosheet materials. The synthesized MFI nanosheet catalysts were fullymore » characterized and evaluated for cellulose pyrolysis vapor upgrading to produce olefins and aromatic hydrocarbons. The data obtained from pyrolysis-GCMS (py-GCMS), showed that fresh MFI nanosheets produced similar aromatic hydrocarbon and olefin yields compared to conventional HZSM-5. However, MFI nanosheets demonstrated a longer lifetime than HZSM-5 even though coke contents were also higher than for HZSM-5 because the mesopores enabled better accessibility to active acid sites. This conclusion was supported by results from post-reaction analysis of various spent catalysts collected at different points during the deactivation experiments.« less

Conception and construction of an LPG tank using a composite membrane technology

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

Fuvel, P.; Claude, J.

TECHNIGAZ and TOTAL C.F.P. have developed a new LPG storage technology derived from the membrane concept used for LNG storage and transportation. This technology called GMS uses a composite membrane as primary barrier. A 2 000 m/sup 3/ storage pilot unit, based on that concept, is under construction in TOTAL's refinery at DUNKIRK (France) since September 1983.

Generalised Spin Dynamics and Induced Bounds of Automorphic [A]nX, [AX]n NMR Systems via Dual Tensorial Sets: An Invariant Cardinality Role for CFP

NASA Astrophysics Data System (ADS)

Temme, Francis P.

For uniform spins and their indistinguishable point sets of tensorial bases defining automorphic group-based Liouvillian NMR spin dynamics, the role of recursively-derived coefficients of fractional parentage (CFP) bijections and Schur duality-defined CFP(0)(n) ≡ ¦GI¦(n) group invariant cardinality is central both to understanding the impact of time-reversal invariance(TRI) spin physics, and to analysis as density-matrix formalisms over democratic recoupled (DR) dual tensorial sets, {T{ṽ}k(11.1)(SU2 × ln)}. Over abstract spin space, these tensorial sets are (ṽ) invariant-theoretic forms which lie beyond the Liouvillian graph recoupling and Racah-forms envisaged by Sanctuary [1]. This is a direct consequence of the dominance of the ln group. It leads to new views on the value of projective group actions as mappings over specialised Liouvillian carrier spaces, and on the need for the replacement of Racah-Wigner (R-W) orthogonality for distinct point sets, by criteria based on explicit properties of invariants [J. Phys.: Math. & Theor. A 41, 015210 (2008)] for multiple invariant systems. Ũ × P group actions over disjoint (L) carrier subspaces, leading to exclusively combinatorial views of the nature of quantal completeness for indistinguishable point-based tensorial sets. Such generalised invariant-theoretic approaches lie beyond the range of Lévi-Civitá generator views, or of Lévy-Leblond and Lévy-Nahas [9] with its additional cyclic-commutators defining mono-invariant DR forms. Comparison of the latter with generalised multiple-invariant techniques provides an answer to the question of precisely why [A]n≥4(X) and [AX]n≥4 NMR system spin dynamics are not ameniable to conventional R-W analysis of recoupled discrete-point tensorial systems. Our work augments earlier Hilbert space views, both of Louck and Biedenharn [21] on boson pattern projective mapping, and of Corio [19]. The roles of recent ln group action and (λ ⊢ n)-Schur combinatorial concepts, as well as of polyhedral-combinatorial modelling over invariance algebras, contribute significantly to our understanding of invariant-based techniques of Liouville dual tensorial sets for automorphic NMR spin physics.1

Contribution to the risk characterization of ciguatoxins: LOAEL estimated from eight ciguatera fish poisoning events in Guadeloupe (French West Indies)

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

Hossen, Virginie; Soliño, Lucia; Leroy, Patricia

From 2010 to 2012, 35 ciguatera fish poisoning (CFP) events involving 87 individuals who consumed locally-caught fish were reported in Guadeloupe (French West Indies). For 12 of these events, the presence of ciguatoxins (CTXs) was indicated in meal remnants and in uncooked fish by the mouse bioassay (MBA). Caribbean ciguatoxins (C-CTXs) were confirmed by liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis. Using a cell-based assay (CBA), and the only available standard Pacific ciguatoxin-1 (P-CTX-1), the lowest toxins level detected in fish samples causing CFP was 0.022 µg P-CTX-1 equivalent (eq.)·kg{sup −1} fish. Epidemiological and consumption data were compiled for most ofmore » the individuals afflicted, and complete data for establishing the lowest observable adverse effects level (LOAEL) were obtained from 8 CFP events involving 21 individuals. Based on toxin intakes, the LOAEL was estimated at 4.2 ng P-CTX-1 eq./individual corresponding to 48.4 pg P-CTX-1 eq. kg{sup −1} body weight (bw). Although based on limited data, these results are consistent with the conclusions of the European Food Safety Authority (EFSA) opinion which indicates that a level of 0.01 µg P-CTX-1 eq. kg{sup −1} fish, regardless of source, should not exert effects in sensitive individuals when consuming a single meal. The calculated LOAEL is also consistent with the U.S. Food and Drug Administration guidance levels for CTXs (0.1 µg C-CTX-1 eq. kg{sup −1} and 0.01 µg P-CTX-1 eq. kg{sup −1} fish). - Highlights: • We report on an epidemiological study on Ciguatera events in the French West Indies. • The collection of consumption data allows for the first time the LOAEL determination. • The LOAEL for ciguatoxins was established at 48.4 pg P-CTX-1 eq. kg{sup −1} bw. • LC–MS/MS provided structural confirmation of C-CTX1 in two suspected samples • Neuro-2A CBA is suitable for assessing composite toxicity levels in fish samples.« less

Flow Cytometry Enables Multiplexed Measurements of Genetically Encoded Intramolecular FRET Sensors Suitable for Screening.

PubMed

Doucette, Jaimee; Zhao, Ziyan; Geyer, Rory J; Barra, Melanie M; Balunas, Marcy J; Zweifach, Adam

2016-07-01

Genetically encoded sensors based on intramolecular FRET between CFP and YFP are used extensively in cell biology research. Flow cytometry has been shown to offer a means to measure CFP-YFP FRET; we suspected it would provide a unique way to conduct multiplexed measurements from cells expressing different FRET sensors, which is difficult to do with microscopy, and that this could be used for screening. We confirmed that flow cytometry accurately measures FRET signals using cells transiently transfected with an ERK activity reporter, comparing responses measured with imaging and cytometry. We created polyclonal long-term transfectant lines, each expressing a different intramolecular FRET sensor, and devised a way to bar-code four distinct populations of cells. We demonstrated the feasibility of multiplexed measurements and determined that robust multiplexed measurements can be conducted in plate format. To validate the suitability of the method for screening, we measured responses from a plate of bacterial extracts that in unrelated experiments we had determined contained the protein kinase C (PKC)-activating compound teleocidin A-1. The multiplexed assay correctly identifying the teleocidin A-1-containing well. We propose that multiplexed cytometric FRET measurements will be useful for analyzing cellular function and for screening compound collections. © 2016 Society for Laboratory Automation and Screening.

Effects of turbulence on hydraulic heads and parameter sensitivities in preferential groundwater flow layers

USGS Publications Warehouse

Shoemaker, W. Barclay; Cunningham, Kevin J.; Kuniansky, Eve L.; Dixon, Joann F.

2008-01-01

A conduit flow process (CFP) for the Modular Finite Difference Ground‐Water Flow model, MODFLOW‐2005, has been created by the U.S. Geological Survey. An application of the CFP on a carbonate aquifer in southern Florida is described; this application examines (1) the potential for turbulent groundwater flow and (2) the effects of turbulent flow on hydraulic heads and parameter sensitivities. Turbulent flow components were spatially extensive in preferential groundwater flow layers, with horizontal hydraulic conductivities of about 5,000,000 m d−1, mean void diameters equal to about 3.5 cm, groundwater temperature equal to about 25°C, and critical Reynolds numbers less than or equal to 400. Turbulence either increased or decreased simulated heads from their laminar elevations. Specifically, head differences from laminar elevations ranged from about −18 to +27 cm and were explained by the magnitude of net flow to the finite difference model cell. Turbulence also affected the sensitivities of model parameters. Specifically, the composite‐scaled sensitivities of horizontal hydraulic conductivities decreased by as much as 70% when turbulence was essentially removed. These hydraulic head and sensitivity differences due to turbulent groundwater flow highlight potential errors in models based on the equivalent porous media assumption, which assumes laminar flow in uniformly distributed void spaces.

The Epiphytic Genus Gambierdiscus (Dinophyceae) in the Kermadec Islands and Zealandia Regions of the Southwestern Pacific and the Associated Risk of Ciguatera Fish Poisoning.

PubMed

Rhodes, Lesley L; Smith, Kirsty F; Murray, Sam; Harwood, D Tim; Trnski, Tom; Munday, Rex

2017-07-11

Species in the genus Gambierdiscus produce ciguatoxins (CTXs) and/or maitotoxins (MTXs), which may cause ciguatera fish poisoning (CFP) in humans if contaminated fish are consumed. Species of Gambierdiscus have previously been isolated from macroalgae at Rangitahua (Raoul Island and North Meyer Islands, northern Kermadec Islands), and the opportunity was taken to sample for Gambierdiscus at the more southerly Macauley Island during an expedition in 2016. Gambierdiscus cells were isolated, cultured, and DNA extracted and sequenced to determine the species present. Bulk cultures were tested for CTXs and MTXs by liquid chromatography-mass spectrometry (LC-MS/MS). The species isolated were G. australes , which produced MTX-1 (ranging from 3 to 36 pg/cell), and G. polynesiensis , which produced neither MTX-1 nor, unusually, any known CTXs. Isolates of both species produced putative MTX-3. The risk of fish, particularly herbivorous fish, causing CFP in the Zealandia and Kermadec Islands region is real, although in mainland New Zealand the risk is currently low. Both Gambierdiscus and Fukuyoa have been recorded in the sub-tropical northern region of New Zealand, and so the risk may increase with warming seas and shift in the distribution of Gambierdiscus species.

The Epiphytic Genus Gambierdiscus (Dinophyceae) in the Kermadec Islands and Zealandia Regions of the Southwestern Pacific and the Associated Risk of Ciguatera Fish Poisoning

PubMed Central

Rhodes, Lesley L.; Smith, Kirsty F.; Murray, Sam; Harwood, D. Tim; Trnski, Tom; Munday, Rex

2017-01-01

Species in the genus Gambierdiscus produce ciguatoxins (CTXs) and/or maitotoxins (MTXs), which may cause ciguatera fish poisoning (CFP) in humans if contaminated fish are consumed. Species of Gambierdiscus have previously been isolated from macroalgae at Rangitahua (Raoul Island and North Meyer Islands, northern Kermadec Islands), and the opportunity was taken to sample for Gambierdiscus at the more southerly Macauley Island during an expedition in 2016. Gambierdiscus cells were isolated, cultured, and DNA extracted and sequenced to determine the species present. Bulk cultures were tested for CTXs and MTXs by liquid chromatography-mass spectrometry (LC-MS/MS). The species isolated were G. australes, which produced MTX-1 (ranging from 3 to 36 pg/cell), and G. polynesiensis, which produced neither MTX-1 nor, unusually, any known CTXs. Isolates of both species produced putative MTX-3. The risk of fish, particularly herbivorous fish, causing CFP in the Zealandia and Kermadec Islands region is real, although in mainland New Zealand the risk is currently low. Both Gambierdiscus and Fukuyoa have been recorded in the sub-tropical northern region of New Zealand, and so the risk may increase with warming seas and shift in the distribution of Gambierdiscus species. PMID:28696400

Expression and Secretion of Cyan Fluorescent Protein (CFP) in B. subtilis using the Chitinase Promoter from Bacillus pumilus SG2

PubMed Central

Shali, Abbas; Rigi, Garshasb; Pornour, Majid; Ahmadian, Gholamreza

2017-01-01

Background: Improved cyan fluorescent protein (ICFP) is a monochromic, green fluorescent protein (GFP) derivative produced by Aequorea macrodactyla in a process similar to GFP. This protein has strong absorption spectra at wavelengths 426-446 nm. ICFP can be used in cell, organelle or intracellular protein labeling, investigating the protein-protein interactions as well as assessing the promoter activities. Methods: In our previous study, the promoters of two chitinases (ChiS and ChiL) from Bacillus pumilus SG2 were assessed in B. subtilis and their regulatory elements were characterized. In the present study, icfp was cloned downstream of several truncated promoters obtained in the former study, and ICFP expression was evaluated in B. subtilis. Results: Extracellular expression and secretion of ICFP were analyzed under the control of different truncated versions of ChiSL promoters grown on different media. Results from SDS-PAGE and fluorimetric analyses showed that there were different expression rates of CFP; however, the UPChi-ICFP3 construct exhibited a higher level of expression and secretion in the culture medium. Conclusion: Our presented results revealed that inserting this truncated form of Chi promoter upstream of the ICFP, as a reporter gene, in B. subtilis led to an approximately ten fold increase in ICFP expression. PMID:28088132

Community food program use in Inuvik, Northwest Territories

PubMed Central

2013-01-01

Background Community food programs (CFPs) provide an important safety-net for highly food insecure community members in the larger settlements of the Canadian Arctic. This study identifies who is using CFPs and why, drawing upon a case study from Inuvik, Northwest Territories. This work is compared with a similar study from Iqaluit, Nunavut, allowing the development of an Arctic-wide understanding of CFP use – a neglected topic in the northern food security literature. Methods Photovoice workshops (n=7), a modified USDA food security survey and open ended interviews with CFP users (n=54) in Inuvik. Results Users of CFPs in Inuvik are more likely to be housing insecure, female, middle aged (35–64), unemployed, Aboriginal, and lack a high school education. Participants are primarily chronic users, and depend on CFPs for regular food access. Conclusions This work indicates the presence of chronically food insecure groups who have not benefited from the economic development and job opportunities offered in larger regional centers of the Canadian Arctic, and for whom traditional kinship-based food sharing networks have been unable to fully meet their dietary needs. While CFPs do not address the underlying causes of food insecurity, they provide an important service for communities undergoing rapid change, and need greater focus in food policy herein. PMID:24139485

Invasive Lionfish (Pterois volitans): A Potential Human Health Threat for Ciguatera Fish Poisoning in Tropical Waters

PubMed Central

Robertson, Alison; Garcia, Ana C.; Flores Quintana, Harold A.; Smith, Tyler B.; Castillo, Bernard F.; Reale-Munroe, Kynoch; Gulli, Joseph A.; Olsen, David A.; Hooe-Rollman, Jennifer I.; Jester, Edward L. E.; Klimek, Brian J.; Plakas, Steven M.

2013-01-01

Invasive Indo-Pacific lionfish (Pterois volitans) have rapidly expanded in the Western Atlantic over the past decade and have had a significant negative impact on reef fish biodiversity, habitat, and community structure, with lionfish out-competing native predators for resources. In an effort to reduce this population explosion, lionfish have been promoted for human consumption in the greater Caribbean region. This study examined whether the geographical expansion of the lionfish into a known ciguatera-endemic region can pose a human health threat for ciguatera fish poisoning (CFP). More than 180 lionfish were collected from waters surrounding the US Virgin Islands throughout 2010 and 2011. Ciguatoxin testing included an in vitro neuroblastoma cytotoxicity assay for composite toxicity assessment of sodium-channel toxins combined with confirmatory liquid chromatography tandem mass spectrometry. A 12% prevalence rate of ciguatoxic lionfish exceeding the FDA guidance level of 0.1 µg/kg C-CTX-1 equivalents was identified in fish from the U.S. Virgin Islands, highlighting a potential consumption risk in this region. This study presents the first evidence that the invasive lionfish, pose a direct human health risk for CFP and highlights the need for awareness and research on this food safety hazard in known endemic areas. PMID:24378919

Invasive lionfish (Pterois volitans): a potential human health threat for ciguatera fish poisoning in tropical waters.

PubMed

Robertson, Alison; Garcia, Ana C; Quintana, Harold A Flores; Smith, Tyler B; Castillo, Bernard F; Reale-Munroe, Kynoch; Gulli, Joseph A; Olsen, David A; Hooe-Rollman, Jennifer I; Jester, Edward L E; Klimek, Brian J; Plakas, Steven M

2013-12-27

Invasive Indo-Pacific lionfish (Pterois volitans) have rapidly expanded in the Western Atlantic over the past decade and have had a significant negative impact on reef fish biodiversity, habitat, and community structure, with lionfish out-competing native predators for resources. In an effort to reduce this population explosion, lionfish have been promoted for human consumption in the greater Caribbean region. This study examined whether the geographical expansion of the lionfish into a known ciguatera-endemic region can pose a human health threat for ciguatera fish poisoning (CFP). More than 180 lionfish were collected from waters surrounding the US Virgin Islands throughout 2010 and 2011. Ciguatoxin testing included an in vitro neuroblastoma cytotoxicity assay for composite toxicity assessment of sodium-channel toxins combined with confirmatory liquid chromatography tandem mass spectrometry. A 12% prevalence rate of ciguatoxic lionfish exceeding the FDA guidance level of 0.1 µg/kg C-CTX-1 equivalents was identified in fish from the U.S. Virgin Islands, highlighting a potential consumption risk in this region. This study presents the first evidence that the invasive lionfish, pose a direct human health risk for CFP and highlights the need for awareness and research on this food safety hazard in known endemic areas.

Overview of MPEG internet video coding

NASA Astrophysics Data System (ADS)

Wang, R. G.; Li, G.; Park, S.; Kim, J.; Huang, T.; Jang, E. S.; Gao, W.

2015-09-01

MPEG has produced standards that have provided the industry with the best video compression technologies. In order to address the diversified needs of the Internet, MPEG issued the Call for Proposals (CfP) for internet video coding in July, 2011. It is anticipated that any patent declaration associated with the Baseline Profile of this standard will indicate that the patent owner is prepared to grant a free of charge license to an unrestricted number of applicants on a worldwide, non-discriminatory basis and under other reasonable terms and conditions to make, use, and sell implementations of the Baseline Profile of this standard in accordance with the ITU-T/ITU-R/ISO/IEC Common Patent Policy. Three different codecs had responded to the CfP, which are WVC, VCB and IVC. WVC was proposed jointly by Apple, Cisco, Fraunhofer HHI, Magnum Semiconductor, Polycom and RIM etc. it's in fact AVC baseline. VCB was proposed by Google, and it's in fact VP8. IVC was proposed by several Universities (Peking University, Tsinghua University, Zhejiang University, Hanyang University and Korea Aerospace University etc.) and its coding tools was developed from Zero. In this paper, we give an overview of the coding tools in IVC, and evaluate its performance by comparing it with WVC, VCB and AVC High Profile.

Recombinant bacille Calmette-Guerin coexpressing Ag85b, CFP10, and interleukin-12 elicits effective protection against Mycobacterium tuberculosis.

PubMed

Chen, Yih-Yuan; Lin, Chih-Wei; Huang, Wei-Feng; Chang, Jia-Ru; Su, Ih-Jen; Hsu, Chih-Hao; Cheng, Han-Yin; Hsu, Shu-Ching; Dou, Horng-Yunn

2017-02-01

The tuberculosis (TB) pandemic remains a leading cause of human morbidity and mortality, despite widespread use of the only licensed anti-TB vaccine, bacille Calmette-Guerin (BCG). The protective efficacy of BCG in preventing pulmonary TB is highly variable; therefore, an effective new vaccine is urgently required. In the present study, we assessed the ability of novel recombinant BCG vaccine (rBCG) against Mycobacterium tuberculosis by using modern immunological methods. Enzyme-linked immunospot assays demonstrated that the rBCG vaccine, which coexpresses two mycobacterial antigens (Ag85B and CFP10) and human interleukin (IL)-12 (rBCG2) elicits greater interferon-γ (IFN-γ) release in the mouse lung and spleen, compared to the parental BCG. In addition, rBCG2 triggers a Th1-polarized response. Our results also showed that rBCG2 vaccination significantly limits M. tuberculosis H37Rv multiplication in macrophages. The rBCG2 vaccine surprisingly induces significantly higher tumor necrosis factor-α (TNF-α) production by peripheral blood mononuclear cells that were exposed to a nonmycobacterial stimulus, compared to the parental BCG. In this study, we demonstrated that the novel rBCG2 vaccine may be a promising candidate vaccine against M. tuberculosis infection. Copyright © 2014. Published by Elsevier B.V.

Nonenzymatic glucose sensor based on renewable electrospun Ni nanoparticle-loaded carbon nanofiber paste electrode.

PubMed

Liu, Yang; Teng, Hong; Hou, Haoqing; You, Tianyan

2009-07-15

A novel nonenzymatic glucose sensor was developed based on the renewable Ni nanoparticle-loaded carbon nanofiber paste (NiCFP) electrode. The NiCF nanocomposite was prepared by combination of electrospinning technique with thermal treatment method. The scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images showed that large amounts of spherical nanoparticles were well dispersed on the surface or embedded in the carbon nanofibers. And the nanoparticles were composed of Ni and NiO, as revealed by energy dispersive X-ray spectroscopy (EDX) and X-ray powder diffraction (XRD). In application to nonenzymatic glucose determination, the renewable NiCFP electrodes, which were constructed by simply mixing the electrospun nanocomposite with mineral oil, exhibited strong and fast amperometric response without being poisoned by chloride ions. Low detection limit of 1 microM with wide linear range from 2 microM to 2.5 mM (R=0.9997) could be obtained. The current response of the proposed glucose sensor was highly sensitive and stable, attributing to the electrocatalytic performance of the firmly embedded Ni nanoparticles as well as the chemical inertness of the carbon-based electrode. The good analytical performance, low cost and straightforward preparation method made this novel electrode material promising for the development of effective glucose sensor.

Spatio-temporal imaging of EGF-induced activation of protein kinase A by FRET in living cells

NASA Astrophysics Data System (ADS)

Wang, Jin Jun; Chen, Xiao-Chuan; Xing, Da

2004-07-01

Intracellular molecular interaction is important for the study of cell physiology, yet current relevant methods require fixation or microinjection and lack temporal or spatial resolution. We introduced a new method -- fluorescence resonance energy transfer (FRET) to detect molecular interaction in living cells. On the basis of FRET principle, A-kinase activity reporter (AKAR) protein was designed to consist of the fusions of cyan fluorescent protein (CFP), a phosphoamino acid binding domain, a consensus substrate for protein kinase-A (PKA), and yellow fluorescent protein (YFP). In this study, the designed pAKAR plasmid was used to transfect a human lung cancer cell line (ASTC-a-1). When the AKAR-transfected cells were treated by forskolin (Fsk), we were able to observe the efficient transfer of energy from excited CFP to YFP within the AKAR molecule by fluorescence microcopy, whereas no FRET was detected in the transfected cells without the treatment of Fsk. When the cells were treated by Epidermal growth factor (EGF), the change of FRET was observed at different subcellular locations, reflecting PKA activation inside the cells upon EGF stimulation. The successful design of a fluorescence reporter of PKA activation and its application demonstrated the superiority of this technology in the research of intracellular protein-protein interaction.

Measurement of caspase-2 activation during different anti-tumor drugs induced apoptosis by FRET technique

NASA Astrophysics Data System (ADS)

Lin, Juqiang; Zeng, Shaoqun; Luo, Qingming; Rong, Chen; Zhang, Zhihong

2007-11-01

Caspase-2 is important for the engagement of the mitochondrial apoptotic pathway, in the presence of DNA-damaging agents, such as cisplatin; however, the mechanism by which caspase-2 executes apoptosis remains obscure. In this study, we carried out the measurements of the dynamics of caspase-2 activation in a single living cell by a FRET (fluorescence resonance energy transfer) probe. A FRET probe was constructed that encoded a CRS (caspase-2 recognition site) fused with a cyan fluorescent protein (CFP) and a red fluorescent protein (DsRed) (CFP-CRS-DsRed). Using this probe, we found that during TRAIL-induced apoptosis, caspase-2 was not activated, and caspase-2 activation occurred in etoposide and cisplatin treated cells. However, during cisplatin-induced apoptosis caspase-2 activation was initiated much earlier than that of etoposide. Cisplatin and etoposide is one of the most broadly used drugs in the Clinical applications of cancer chemotherapy, and TRAIL, which belongs to the TNF family proteins, can selectively induce apoptosis in many transformed cells but not in normal cells. Most of anticancer drugs can induce apoptosis mediated by the activation of caspase pathway. Thus, the perfect synergistic effect group of multi-drug can be selected by using our FRET probe.

Investigating MUC1/ICAM-1 Binding Induced Signaling in Breast Cancer Metastasis

DTIC Science & Technology

2011-05-01

expected that covalently linked species would remain intact. Reducing (R, + !-mercaptoethanol) and non-reducing (NR, no !-mercaptoethanol) samples were...binding site, containing both proline and arginine residues. We mutated the SH2 and/or putative SH3 binding domains on the MUC1-CFP-Fv plasmid...Structure and regulation of Src family kinases. Oncogene 2004, 23:7918- 7927. 31. Li SSC: Specificity and versatility of SH3 and other proline -recognition

Ab Initio Surface Phase Diagrams for Coadsorption of Aromatics and Hydrogen on the Pt(111) Surface

DOE PAGES

Ferguson, Glen Allen; Vorotnikov, Vassili; Wunder, Nicholas; ...

2016-11-02

Supported metal catalysts are commonly used for the hydrogenation and deoxygenation of biomass-derived aromatic compounds in catalytic fast pyrolysis. To date, the substrate-adsorbate interactions under reaction conditions crucial to these processes remain poorly understood, yet understanding this is critical to constructing detailed mechanistic models of the reactions important to catalytic fast pyrolysis. Density functional theory (DFT) has been used in identifying mechanistic details, but many of these works assume surface models that are not representative of realistic conditions, for example, under which the surface is covered with some concentration of hydrogen and aromatic compounds. In this study, we investigate hydrogen-guaiacolmore » coadsorption on Pt(111) using van der Waals-corrected DFT and ab initio thermodynamics over a range of temperatures and pressures relevant to bio-oil upgrading. We find that relative coverage of hydrogen and guaiacol is strongly dependent on the temperature and pressure of the system. Under conditions relevant to ex situ catalytic fast pyrolysis (CFP; 620-730 K, 1-10 bar), guaiacol and hydrogen chemisorb to the surface with a submonolayer hydrogen (~0.44 ML H), while under conditions relevant to hydrotreating (470-580 K, 10-200 bar), the surface exhibits a full-monolayer hydrogen coverage with guaiacol physisorbed to the surface. These results correlate with experimentally observed selectivities, which show ring saturation to methoxycyclohexanol at hydrotreating conditions and deoxygenation to phenol at CFP-relevant conditions. Additionally, the vibrational energy of the adsorbates on the surface significantly contributes to surface energy at higher coverage. Ignoring this contribution results in not only quantitatively, but also qualitatively incorrect interpretation of coadsorption, shifting the phase boundaries by more than 200 K and ~10-20 bar and predicting no guaiacol adsorption under CFP and hydrotreating conditions. We discuss the implications of this work in the context of modeling hydrogenation and deoxygenation reactions on Pt(111), and we find that only the models representative of equilibrium surface coverage can capture the hydrogenation kinetics correctly. Lastly, as a major outcome of this work, we introduce a freely available web-based tool, dubbed the Surface Phase Explorer (SPE), which allows researchers to conveniently determine surface composition for any one- or two-component system at thermodynamic equilibrium over a wide range of temperatures and pressures on any crystalline surface using standard DFT output.« less

Live Attenuated Salmonella Vaccines Displaying Regulated Delayed Lysis and Delayed Antigen Synthesis To Confer Protection against Mycobacterium tuberculosis

PubMed Central

Juárez-Rodríguez, María Dolores; Yang, Jiseon; Kader, Rebin; Alamuri, Praveen; Curtiss, Roy

2012-01-01

Live recombinant attenuated Salmonella vaccine (RASV) strains have great potential to induce protective immunity against Mycobacterium tuberculosis by delivering M. tuberculosis antigens. Recently, we reported that, in orally immunized mice, RASV strains delivering the M. tuberculosis early secreted antigenic target 6-kDa (ESAT-6) protein and culture filtrate protein 10 (CFP-10) antigens via the Salmonella type III secretion system (SopE amino-terminal region residues 1 to 80 with two copies of ESAT-6 and one copy of CFP-10 [SopENt80-E2C]) afforded protection against aerosol challenge with M. tuberculosis. Here, we constructed and evaluated an improved Salmonella vaccine against M. tuberculosis. We constructed translational fusions for the synthesis of two copies of ESAT-6 plus CFP-10 fused to the OmpC signal sequence (OmpCSS-E2C) and amino acids 44 to 338 of antigen 85A (Ag85A294) flanked by the signal sequence (SS) and C-terminal peptide (CT) of β-lactamase (BlaSS-Ag85A294-BlaCT) to enable delivery via the Salmonella type II secretion system. The genes expressing these proteins were cloned as an operon transcribed from Ptrc into isogenic Asd+/MurA+ pYA3681 lysis vector derivatives with different replication origins (pBR, p15A, pSC101), resulting in pYA4890, pYA4891, and pYA4892 for SopENt80-E2C/Ag85A294 synthesis and pYA4893 and pYA4894 for OmpCSS-E2C/Ag85A294 synthesis. Mice orally immunized with the RASV χ11021 strain engineered to display regulated delayed lysis and regulated delayed antigen synthesis in vivo and harboring pYA4891, pYA4893, or pYA4894 elicited significantly greater humoral and cellular immune responses, and the RASV χ11021 strain afforded a greater degree of protection against M. tuberculosis aerosol challenge in mice than RASVs harboring any other Asd+/MurA+ lysis plasmid and immunization with M. bovis BCG, demonstrating that RASV strains displaying regulated delayed lysis with delayed antigen synthesis resulted in highly immunogenic delivery vectors for oral vaccination against M. tuberculosis infection. PMID:22144485

Stance control is not affected by paresis and reflex hyperexcitability: the case of spastic patients

PubMed Central

Nardone, A; Galante, M; Lucas, B; Schieppati, M

2001-01-01

OBJECTIVES—Spastic patients were studied to understand whether stance unsteadiness is associated with changes in the control of voluntary force, muscle tone, or reflex excitability, rather than to abnormal posture connected to the motor deficit itself.
METHODS—Twenty four normal subjects, 12 patients affected by amyotrophic lateral sclerosis (ALS), seven by spastic paraparesis, and 14 by hemiparesis were studied. All patients featured various degrees of spasticity and paresis but were free from clinically evident sensory deficits. Body sway during quiet upright stance was assessed through a stabilometric platform under both eyes open (EO) and eyes closed (EC) conditions. The sudden rotation of a supporting platform, in a toe up and toe down direction respectively, evoked short (SLR) and medium latency (MLR) reflex responses to stretch of the soleus or the tibialis anterior (TA) muscle.
RESULTS—No relation was found between clinical findings (tone, muscle strength, tendon reflexes, plantar response, and duration of disease) and body sway. On average, all patient groups exhibited a forward shift of the centre of foot pressure (CFP) with respect to normal subjects; in addition, paraparetic and to a much larger extent hemiparetic patients showed a lateral shift of CFP. Body sway area was significantly increased only in the hemiparetic patients. No relation was found between position of the CFP and sway within any patient group. Soleus SLR was increased in all patients with respect to normal subjects. TA SLR was often seen in both patients with ALS and paraparetic patients, but only rarely in normal subjects and hemiparetic patients. However, no relation was found between amplitude of soleus or TA SLRs and stabilometric variables. The frequency and size of soleus MLR and TA MLR were decreased in all patients. These responses were decreased in size and not modulated by background EMG in the affected leg of hemiparetic patients, suggesting a disturbed control of spinal reflexes fed by spindle group II afferent fibres.
CONCLUSIONS—It is proposed that body posture, paresis, or monosynaptic reflex hyperexcitability do not affect the control of equilibrium during quiet upright stance. In hemiparetic patients, the decreased amplitude of MLRs might be the main cause of the large postural instability. The results are congruent with the hypothesis of a role for group II afferent input in the reflex control of equilibrium.

 PMID:11309458

Technical Note: Approximate solution of transient drawdown for constant-flux pumping at a partially penetrating well in a radial two-zone confined aquifer

NASA Astrophysics Data System (ADS)

Huang, C.-S.; Yang, S.-Y.; Yeh, H.-D.

2015-03-01

An aquifer consisting of a skin zone and a formation zone is considered as a two-zone aquifer. Existing solutions for the problem of constant-flux pumping (CFP) in a two-zone confined aquifer involve laborious calculation. This study develops a new approximate solution for the problem based on a mathematical model including two steady-state flow equations with different hydraulic parameters for the skin and formation zones. A partially penetrating well may be treated as the Neumann condition with a known flux along the screened part and zero flux along the unscreened part. The aquifer domain is finite with an outer circle boundary treated as the Dirichlet condition. The steady-state drawdown solution of the model is derived by the finite Fourier cosine transform. Then, an approximate transient solution is developed by replacing the radius of the boundary in the steady-state solution with an analytical expression for a dimensionless time-dependent radius of influence. The approximate solution is capable of predicting good temporal drawdown distributions over the whole pumping period except at the early stage. A quantitative criterion for the validity of neglecting the vertical flow component due to a partially penetrating well is also provided. Conventional models considering radial flow without the vertical component for the CFP have good accuracy if satisfying the criterion.

Development of a Gene Expression Assay for the Diagnosis of Mycobacterium bovis Infection in African Lions (Panthera leo).

PubMed

Olivier, T T; Viljoen, I M; Hofmeyr, J; Hausler, G A; Goosen, W J; Tordiffe, A S W; Buss, P; Loxton, A G; Warren, R M; Miller, M A; van Helden, P D; Parsons, S D C

2017-06-01

Mycobacterium bovis infection, the cause of bovine tuberculosis (BTB), is endemic in wildlife in the Kruger National Park (KNP), South Africa. In lions, a high infection prevalence and BTB mortalities have been documented in the KNP; however, the ecological consequences of this disease are currently unknown. Sensitive assays for the detection of this infection in this species are therefore required. Blood from M. bovis-exposed, M. bovis-unexposed, M. tuberculosis-exposed and M. bovis-infected lions was incubated in QuantiFERON ® -TB Gold (QFT) tubes containing either saline or ESAT-6/CFP-10 peptides. Using qPCR, selected reference genes were evaluated for expression stability in these samples and selected target genes were evaluated as markers of antigen-dependent immune activation. The abundance of monokine induced by gamma interferon (MIG/CXCL9) mRNA, measured in relation to that of YWHAZ, was used as a marker of ESAT-6/CFP-10 sensitization. The gene expression assay results were compared between lion groups, and lenient and stringent diagnostic cut-off values were calculated. This CXCL9 gene expression assay combines a highly specific stimulation platform with a sensitive diagnostic marker that allows for discrimination between M. bovis-infected and M. bovis-uninfected lions. © 2015 Blackwell Verlag GmbH.

FRET-based sensors for the human M1-, M3-, and M5-acetylcholine receptors.

PubMed

Ziegler, Nicole; Bätz, Julia; Zabel, Ulrike; Lohse, Martin J; Hoffmann, Carsten

2011-02-01

Based on the recently developed approach to generate fluorescence resonance energy transfer (FRET)-based sensors to measure GPCR activation, we generated sensor constructs for the human M(1)-, M(3)-, and M(5)-acetylcholine receptor. The receptors were labeled with cyan fluorescent protein (CFP) at their C-terminus, and with fluorescein arsenical hairpin binder (FlAsH) via tetra-cysteine tags inserted in the third intracellular loop. We then measured FRET between the donor CFP and the acceptor FlAsH in living cells and real time. Agonists like acetylcholine, carbachol, or muscarine activate each receptor construct with half-maximal activation times between 60 and 70ms. Removal of the agonist caused the reversal of the signal. Compared with all other agonists, oxotremorine M differed in two major aspects: it caused significantly slower signals at M(1)- and M(5)-acetylcholine receptors and the amplitude of these signals was larger at the M(1)-acetylcholine receptor. Concentration-response curves for the agonists reveal that all agonists tested, with the mentioned exception of oxotremorine M, caused similar maximal FRET-changes as acetylcholine for the M(1)-, M(3)- and M(5)-acetylcholine receptor constructs. Taken together our data support the notion that orthosteric agonists behave similar at different muscarinic receptor subtypes but that kinetic differences can be observed for receptor activation. Copyright © 2010 Elsevier Ltd. All rights reserved.

Optimizing anti-coking abilities of zeolites by ethylene diamine tetraacetie acid modification on catalytic fast pyrolysis of corn stalk

NASA Astrophysics Data System (ADS)

Zhang, Bo; Zhong, Zhaoping; Song, Zuwei; Ding, Kuan; Chen, Paul; Ruan, Roger

2015-12-01

In order to minimize coke yield during biomass catalytic fast pyrolysis (CFP) process, ethylene diamine tetraacetie acid (EDTA) chemical modification method is carried out to selectively remove the external framework aluminum of HZSM-5 catalyst. X-ray diffraction (XRD), nitrogen (N2)-adsorption and ammonia-temperature programmed desorption (NH3-TPD) techniques are employed to investigate the porosity and acidity characteristics of original and modified HZSM-5 samples. Py-GC/MS and thermo-gravimetric analyzer (TGA) experiments are further conducted to explore the catalytic effect of modified HZSM-5 samples on biomass CFP and to verify the positive effect on coke reduction. Results show that EDTA treatment does not damage the crystal structure of HZSM-5 zeolites, but leads to a slight increase of pore volume and pore size. Meanwhile, the elimination of the strong acid peak indicates the dealumination of outer surface of HZSM-5 zeolites. Treatment time of 2 h (labeled EDTA-2H) is optimal for acid removal and hydrocarbon formation. Among all modified catalysts, EDTA-2H performs the best for deacidification and can obviously increase the yields of positive chemical compositions in pyrolysis products. Besides, EDTA modification can improve the anti-coking properties of HZSM-5 zeolites, and EDTA-2H gives rise to the lowest coke yield.

Pacific ciguatoxins in food web components of coral reef systems in the Republic of Kiribati.

PubMed

Mak, Yim Ling; Wai, Tak-Cheung; Murphy, Margaret B; Chan, Wing Hei; Wu, Jia Jun; Lam, James C W; Chan, Leo L; Lam, Paul K S

2013-12-17

Ciguatera fish poisoning (CFP) is a foodborne illness caused by consumption of coral reef fishes contaminated by ciguatoxins (CTXs); of the known CTX congeners, the Pacific ciguatoxins (P-CTXs) are the most toxic. Little is known about the trophodynamics of P-CTXs in coral reef systems. The present study explores the distribution, transfer, and trophic magnification of P-CTX-1, -2, and -3 in coral reef systems with high (ciguatoxic) and low (reference) ciguatoxicity in a CFP-endemic nation by use of liquid chromatography-tandem mass spectrometry (LC-MS/MS). In ciguatoxic coral reef systems, P-CTXs were detected in 54% of herbivorous fishes [total P-CTXs <0.500-1670 pg/g wet weight (ww)], 72% of omnivorous fishes (<0.500-1810 pg/g ww), and 76% of carnivorous fishes (<0.500-69 500 pg/g ww), as well as a lobster ( Panulirus penicillatus ; 2.36 pg/g ww) and an octopus (Octopodidae; 2.56 pg/g ww). The dominant P-CTXs in grazers and piscivorous fishes were P-CTX-2 and -1, respectively. No significant correlation between P-CTX levels and lipid content in three target predatory fishes indicated that accumulation of P-CTXs does not depend on fat content. A weak but significant positive relationship was observed between δ(15)N and P-CTX-1 levels, but further investigation is required to confirm its biomagnification potential.

Genetic and environmental relationships of different measures of individual cheese yield and curd nutrients recovery with coagulation properties of bovine milk.

PubMed

Cecchinato, A; Bittante, G

2016-03-01

The aim of this study was to elucidate the relationships between various cheesemaking-related traits, namely the well-known traditional milk coagulation properties (MCP), the new curd firming and syneresis traits, the cheese yield, and the curd nutrient recoveries or whey losses (all measured at the individual level). Data were obtained from 1,167 Brown Swiss cows reared in 85 herds. A 2-L milk sample was collected once from each animal and assessed for 10 phenotypes related to changes in curd firmness (CF) over time, plus 7 cheesemaking traits. The CF-related traits included 4 traditional single-point lactodynamographic properties [rennet coagulation time (RCT, min); time to a CF of 20mm, min; and the CF 30 and 45 min after rennet addition (a30 and a45, respectively)], 4 parameters used to model the 360 CF data recorded over time for each milk sample [the potential asymptotic CF at infinite time (CFP, mm); the CF instant rate constant, % × min(-1); the syneresis instant rate constant, % × min(-1); and the RCT obtained from modeling individual samples], and 2 traits calculated from individual equations [the maximum CF(CFmax, mm); and the time at CFmax, min]. The cheesemaking traits included 3 cheese yield traits (weights of the fresh curd, curd solids and curd moisture as percent of the weights of the processed milk) and 4 milk nutrient recoveries in the curd (calculated as the percent ratios between a given nutrient in the curd versus that in the processed milk). Bayesian methodology-based multivariate analyses were used to estimate the phenotypic, additive genetic, herd/date, and residual relationships between the aforementioned traits, whereas statistical inferences were based on the marginal posterior distributions of the parameters of concern. The a45, CFP, and CFmax traits were genetically associated with all of the percent cheese yield traits (the additive genetic correlations varied from 0.752 to 0.855 for a45; 0.496 to 0.583 for CFP; and 0.750 to 0.801 for CFmax) and the nutrient recovery traits (additive genetic correlations varied from 0.296 to 0.901 for a45; 0.428 to 0.697 for CFP; and 0.412 to 0.941 for CFmax). Moreover, the nutrient recoveries for fat, solids, and energy exhibited large additive genetic correlations with the other coagulation and curd firming traits. In particular, recovery of protein and fat were found to be powerful instruments for understanding the relationships between milk technological properties and cheese quantity or quality. We observed only weak genetic relationships with the milk quality and MCP traits, suggesting that the highly heritable trait of protein recovery should perhaps be included as a genetic index when seeking to improve cheesemaking efficiency at the population level. In contrast, we found that fat recovery exhibited moderate genetic variation and could be improved through the CF over time traits, especially using those recorded during the late phase of the curd firming process. Moreover, our results demonstrated that the traditional MCP have limited relevance for predicting individual cheese yield. Therefore, their use for this purpose in the dairy industry and breeding programs seems questionable. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Prevalence of Subretinal Drusenoid Deposits in Older Persons with and without Age-Related Macular Degeneration, by Multimodal Imaging.

PubMed

Zarubina, Anna V; Neely, David C; Clark, Mark E; Huisingh, Carrie E; Samuels, Brian C; Zhang, Yuhua; McGwin, Gerald; Owsley, Cynthia; Curcio, Christine A

2016-05-01

To assess the prevalence of subretinal drusenoid deposits (SDD) in older adults with healthy maculas and early and intermediate age-related macular degeneration (AMD) using multimodal imaging. Cross-sectional study. A total of 651 subjects aged ≥60 years enrolled in the Alabama Study of Early Age-Related Macular Degeneration from primary care ophthalmology clinics. Subjects were imaged using spectral domain optical coherence tomography (SD OCT) of the macula and optic nerve head (ONH), infrared reflectance, fundus autofluorescence, and color fundus photographs (CFP). Eyes were assessed for AMD presence and severity using the Age-Related Eye Disease Study (AREDS) 9-step scale. Criteria for SDD presence were identification on ≥1 en face modality plus SD OCT or on ≥2 en face modalities if absent on SD OCT. Subretinal drusenoid deposits were considered present at the person level if present in 1 or both eyes. Prevalence of SDD in participants with and without AMD. Overall prevalence of SDD was 32% (197/611), with 62% (122/197) affected in both eyes. Persons with SDD were older than those without SDD (70.6 vs. 68.7 years, P = 0.0002). Prevalence of SDD was 23% in subjects without AMD and 52% in subjects with AMD (P < 0.0001). Among those with early and intermediate AMD, SDD prevalence was 49% and 79%, respectively. After age adjustment, those with SDD were 3.4 times more likely to have AMD than those without SDD (95% confidence interval, 2.3-4.9). By using CFP only for SDD detection per the AREDS protocol, prevalence of SDD was 2% (12/610). Of persons with SDD detected by SD OCT and confirmed by at least 1 en face modality, 47% (89/190) were detected exclusively on the ONH SD OCT volume. Subretinal drusenoid deposits are present in approximately one quarter of older adults with healthy maculae and in more than half of persons with early to intermediate AMD, even by stringent criteria. The prevalence of SDD is strongly associated with AMD presence and severity and increases with age, and its retinal topography including peripapillary involvement resembles that of rod photoreceptors. Consensus on SDD detection methods is recommended to advance our knowledge of this lesion and its clinical and biologic significance. Copyright © 2016 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.

Prevalence of subretinal drusenoid deposits in older persons with and without age-related macular degeneration, by multimodal imaging

PubMed Central

Zarubina, Anna V.; Neely, David C.; Clark, Mark E.; Huisingh, Carrie E.; Samuels, Brian C.; Zhang, Yuhua; McGwin, Gerald; Owsley, Cynthia; Curcio, Christine A.

2015-01-01

Purpose To assess the prevalence of subretinal drusenoid deposits (SDD) in older adults with healthy maculas and early and intermediate age-related macular degeneration (AMD) using multimodal imaging. Design Cross-sectional study. Participants A total of 651 subjects aged ≥60 years enrolled in the Alabama Study of Early Age-Related Macular Degeneration from primary care ophthalmology clinics. Methods Subjects were imaged using spectral domain optical coherence tomography (SD-OCT) of the macula and optic nerve head (ONH), infrared reflectance, fundus autofluorescence, and color fundus photographs (CFP). Eyes were assessed for AMD presence and severity using the AREDS 9-step scale. Criteria for SDD presence were identification on ≥1 en-face modality plus SD-OCT or on ≥2 en-face modalities if absent on SD-OCT. SDD were considered present at the person-level if present in 1 or both eyes. Main outcomes measures Prevalence of SDD in participants with and without AMD. Results Overall prevalence of SDD was 32% (197/611), with 62% (122/197) affected in both eyes. Persons with SDD were older than those without SDD (70.6 vs. 68.7 years, p =0.0002). Prevalence of SDD was 23% in subjects without AMD and 52% in subjects with AMD (p<0.0001). Among those with early and intermediate AMD, SDD prevalence was 49% and 79%, respectively. After age adjustment, those with SDD were 3.4x more likely to have AMD than those without SDD (95% CI 2.3–4.9). By using CFP only for SDD detection per the AREDS protocol, prevalence of SDD was 2% (12/610). Of persons with SDD detected by SD-OCT and confirmed by at least one en-face modality 47% (89/190) were detected exclusively on the ONH SD-OCT volume. Conclusion SDD are present in approximately one quarter of older adults with healthy maculae and in more than half of persons with early to intermediate AMD, even by stringent criteria. The prevalence of SDD is strongly associated with AMD presence and severity and increases with age, and its retinal topography including peripapillary involvement resembles that of rod photoreceptors. Consensus on SDD detection methods is recommended to advance our knowledge of this lesion and its clinical and biologic significance. PMID:26875000

Estimating ocean-air heat fluxes during cold air outbreaks by satellite

NASA Technical Reports Server (NTRS)

Chou, S. H.; Atlas, D.

1981-01-01

Nomograms of mean column heating due to surface sensible and latent heat fluxes were developed. Mean sensible heating of the cloud free region is related to the cloud free path (CFP, the distance from the shore to the first cloud formation) and the difference between land air and sea surface temperatures, theta sub 1 and theta sub 0, respectively. Mean latent heating is related to the CFP and the difference between land air and sea surface humidities q sub 1 and q sub 0 respectively. Results are also applicable to any path within the cloud free region. Corresponding heat fluxes may be obtained by multiplying the mean heating by the mean wind speed in the boundary layer. The sensible heating estimated by the present method is found to be in good agreement with that computed from the bulk transfer formula. The sensitivity of the solutions to the variations in the initial coastal soundings and large scale subsidence is also investigated. The results are not sensitive to divergence but are affected by the initial lapse rate of potential temperature; the greater the stability, the smaller the heating, other things being equal. Unless one knows the lapse rate at the shore, this requires another independent measurement. For this purpose the downwind slope of the square of the boundary layer height is used, the mean value of which is also directly proportional to the mean sensible heating. The height of the boundary layer should be measurable by future spaceborn lidar systems.

Late-Transition-Metal-Modified β-Mo 2C Catalysts for Enhanced Hydrogenation during Guaiacol Deoxygenation

DOE PAGES

Baddour, Frederick G.; Witte, Vanessa A.; Nash, Connor P.; ...

2017-10-26

Molybdenum carbide has been identified as a promising bifunctional catalyst in the deoxygenation of a variety of pyrolysis vapor model compounds. Although high deoxygenation activity has been demonstrated, complementary hydrogenation activity has been limited, especially for lignin-derived, aromatic model compounds. The ability to control the relative site densities of acidic and hydrogenation functionalities represents a catalyst design challenge for these materials with the goal to improve hydrogenation activity under ex situ catalytic fast pyrolysis (CFP) conditions. Here in this paper, we demonstrate that the addition of Pt and Ni to Mo 2C resulted in an increase in the H*-site densitymore » with only a minor decrease in the acid-site density. In contrast, the addition of Pd did not significantly alter the H* or acid site densities. High conversions (>94%) and high selectivities to 0-oxygen products (>80%) were observed in guaiacol deoxygenation under ex situ CFP conditions (350 °C and 0.44 MPa H 2) for all catalysts. Pt addition resulted in the greatest deoxygenation, and site-time yields to hydrogenated products over the Pt/Mo 2C catalyst were increased to 0.048 s -1 compared to 0.015-0.019 s -1 for all other catalysts. The Pt/Mo 2C catalyst demonstrated the highest hydrogenation performance, but modification with Ni also significantly enhanced hydrogenation performance, representing a promising lower-cost alternative.« less

Expansion of Pathogen-Specific Mono- and Multifunctional Th1 and Th17 Cells in Multi-Focal Tuberculous Lymphadenitis

PubMed Central

Kumar, Nathella Pavan; Sridhar, Rathinam; Banurekha, Vaithilingam V.; Nair, Dina; Jawahar, Mohideen S.; Nutman, Thomas B.; Babu, Subash

2013-01-01

Background Th1 and Th17 responses are known to play an important role in immunity to pulmonary tuberculosis (PTB), although little is known about their role in extrapulmonary forms of tuberculosis (TB). Methods To identify the role of Th1, Th17, and Th22 cells in multi-focal TB lymphadenitis (TBL), we examined mycobacteria–specific immune responses in the whole blood of individuals with PTB (n = 20) and compared them with those with TBL (n = 25). Results Elevated frequencies of CD4+ T cells expressing IFN- γ, TNF-α, and IL-2 were present in individuals with TBL compared with those with PTB at baseline and in response to ESAT-6 and CFP-10. Similarly, increased frequencies of CD4+ T cells expressing IL-17A, IL-17F, and IFN-γ were also present in individuals with TBL at baseline and following ESAT-6 and CFP-10 stimulation although no significant difference in frequency of Th22 cells was observed. Finally, frequencies of Th1 (but not Th17) cells exhibited a significantly negative correlation with natural regulatory T cell frequencies at baseline. Conclusions Multi-focal TB lymphadenitis is therefore characterized by elevated frequencies of Th1 and Th17 cells, indicating that Th1 and Th17 responses in TB disease are probably correlates of disease severity rather than of protective immunity. PMID:23451159

Late-Transition-Metal-Modified β-Mo 2C Catalysts for Enhanced Hydrogenation during Guaiacol Deoxygenation

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

Baddour, Frederick G.; Witte, Vanessa A.; Nash, Connor P.

Molybdenum carbide has been identified as a promising bifunctional catalyst in the deoxygenation of a variety of pyrolysis vapor model compounds. Although high deoxygenation activity has been demonstrated, complementary hydrogenation activity has been limited, especially for lignin-derived, aromatic model compounds. The ability to control the relative site densities of acidic and hydrogenation functionalities represents a catalyst design challenge for these materials with the goal to improve hydrogenation activity under ex situ catalytic fast pyrolysis (CFP) conditions. Here in this paper, we demonstrate that the addition of Pt and Ni to Mo 2C resulted in an increase in the H*-site densitymore » with only a minor decrease in the acid-site density. In contrast, the addition of Pd did not significantly alter the H* or acid site densities. High conversions (>94%) and high selectivities to 0-oxygen products (>80%) were observed in guaiacol deoxygenation under ex situ CFP conditions (350 °C and 0.44 MPa H 2) for all catalysts. Pt addition resulted in the greatest deoxygenation, and site-time yields to hydrogenated products over the Pt/Mo 2C catalyst were increased to 0.048 s -1 compared to 0.015-0.019 s -1 for all other catalysts. The Pt/Mo 2C catalyst demonstrated the highest hydrogenation performance, but modification with Ni also significantly enhanced hydrogenation performance, representing a promising lower-cost alternative.« less

Evaluation of a New IFN-γ Release Assay for Rapid Diagnosis of Active Tuberculosis in a High-Incidence Setting.

PubMed

Li, Gen; Li, Feng; Zhao, Hui-Min; Wen, Han-Li; Li, Hai-Cong; Li, Chun-Ling; Ji, Ping; Xu, Peng; Wu, Kang; Hu, Zhi-Dong; Lu, Shui-Hua; Lowrie, Douglas B; Lv, Jian-Xin; Fan, Xiao-Yong

2017-01-01

Blood-based interferon-gamma (IFN-γ) release assays (IGRAs) have been proven to be useful in the diagnosis of Mycobacterium tuberculosis ( Mtb ) infection. However, IGRAs have not been recommended for clinical practice in most low-income settings due to cost-intensive limitations and shortage of clinical data available. The established T-SPOT. TB assay containing Mtb -specific antigens ESAT-6 and CFP10 are widely used for immunodiagonsis of Mtb infection, but the high cost is one of the restricting factors against its clinical application in the developing countries. More recently, a cost-saving IGRA assay, TS-SPOT, was approved in China. This new assay contains an additional antigen Rv3615c. Rv3615c contains broadly recognized CD4 + and CD8 + epitopes, and T-cell responses to Rv3615c are as specific for Mtb infection as the responses to ESAT-6 and CFP10 in both Mtb -infected humans and M. bovis -infected cattle. Therefore, we assessed the likely effect of inclusion of Rv3615c as stimulus besides ESAT-6 and CFP10 in an IGRA assay and evaluated the performance of TS-SPOT for diagnosis of Mtb infection and active TB compared with T-SPOT. TB . We tested 155 active TB patients, 90 non-TB lung disease patients, and 55 healthy individuals. The results presented an improved positive rate for diagnosis of active TB and Mtb infection, that could be attributable to inclusion of Rv3615c in the mixture of stimulatory antigens. The diagnostic efficiency of TS-SPOT assay for active TB was as follows: sensitivity 80.00%, specificity 83.45%, positive predictive value (PPV) 83.78%, negative predictive value (NPV) 83.45%, positive likelihood ratio (LR+) 4.83, and negative likelihood ratio (LR-) 0.24. The results were similar to those of T-SPOT. TB , with an excellent agreement (κ = 0.91, 95% CI: 0.85-0.95) being observed between these two assays. The sensitivities of the TS-SPOT assay varied for patients with different forms of active TB, with the highest sensitivity for patients with culture-positive pulmonary TB (92.16%) and the lowest for those with tuberculosis meningitis (50.00%). Taken together, the current evidence indicates that this new TS-SPOT assay is a useful adjunct to the current tests for rapid diagnosis of active TB and Mtb infection in low-income and high-incidence settings due to its characteristics of cost-effectiveness and high-quality.

Cytokine response to selected MTB antigens in Ghanaian TB patients, before and at 2 weeks of anti-TB therapy is characterized by high expression of IFN-γ and Granzyme B and inter- individual variation.

PubMed

Mensah, Gloria Ivy; Addo, Kennedy Kwasi; Tetteh, John Amissah; Sowah, Sandra; Loescher, Thomas; Geldmacher, Christof; Jackson-Sillah, Dolly

2014-09-10

There has been a long held belief that patients with drug-susceptible TB are non-infectious after two weeks of therapy. Recent microbiological and epidemiological evidence has challenged this dogma, however, the nature of the Mtb-specific cellular immune response during this period has not been adequately investigated. This knowledge could be exploited in the development of immunological biomarkers of early treatment response. Cellular response to four Mtb infection phase-dependent antigens, ESAT-6/CFP-10 fusion protein and three DosR encoded proteins (Rv1733c, Rv2029c, Rv2628) were evaluated in a Ghanaian TB cohort (n=20) before and after 2 weeks of anti TB therapy. After 6-days in vitro stimulation, Peripheral blood mononuclear cell (PBMC) culture supernatant was harvested and the concentration of IFN-γ, Granzyme B, IL-10, IL-17, sIL2Rα and TNF-α were determined in a 6-plex Luminex assay. Frequencies of IFN-γ + CD4 and CD8 T cells were also determined in an intracellular cytokine assay. All antigens induced higher levels of IFN-γ, followed by Granzyme B, TNF-α and IL-17 and low levels of IL-10 and sIL-2R-α in PBMC before treatment and after 2 weeks of treatment. Median cytokine levels of IFN-γ, Granzyme B, IL-17 and sIL-2R-α increased during week two, but it was significant for only Rv1733-specific production of Granzyme B (P = 0. 013). The median frequency of antigen specific IFN-γ + CD4 T cells increased at week two; however, only the increase in the ESAT-6/CFP-10-specific response was significant (P = 0. 0008). In contrast, the median frequency of ESAT-6/CFP-10- specific IFN-γ + CD8 T cell responses declined during week two (P = 0. 0024). Additionally, wide inter-individual variation with three distinct patterns were observed; increase in all cytokine levels, decrease in all cytokine levels and fluctuating cytokine levels after 2 weeks of treatment. The second week of effective chemotherapy was characterized by a general increase in cytokine response to Mtb-specific antigens suggestive of an improvement in cellular response with therapy. However, the wide inter-individual variation observed would limit the utility of cytokine biomarkers during this period.

Opportunity to Participate in ESSE 21: The 2003 Call for Participation

NASA Astrophysics Data System (ADS)

Ruzek, M.; Johnson, D. R.

2003-12-01

Earth System Science Education for the 21st Century (ESSE 21), sponsored by NASA through the Universities Space Research Association (USRA), is a collaborative undergraduate/graduate education program offering small grants to colleges and universities to engage a diverse interdisciplinary community of faculty and scientists in the development of courses, curricula and degree programs and sharing of learning resources focused on the fundamental understanding and application of Earth system principles for the classroom and laboratory. Through an expanded focus including partnerships with minority institutions, ESSE 21 is further developing broadly based courses, educational resources, electronic learning materials and degree programs that extend Earth system science concepts in both undergraduate and graduate classrooms and laboratories. These resources emphasizing the fundamentals of Earth system science advance the nation's broader agenda for improving science, technology, engineering and mathematics competency. The thrust to establish Earth system and global change science within the classrooms of colleges and universities is critical to laying and extending the foundation for knowledge-based decision making in the 21st century by both scientists and society in an effort to achieve sustainability. ESSE 21 released a Call for Participation (CFP) in the Fall of 2002 soliciting proposals from undergraduate institutions to create and adopt undergraduate and graduate level Earth system science content in courses, curricula and degree programs. In February 2003, twelve college and university teams were competitively selected through the CFP as the Year 1 and Year 2 Program participants. Eight of the participating teams are from minority institutions. The goal for all is to effect systemic change through developing Earth system science learning materials, courses, curricula, degree tracks or programs, and departments that are self-sustaining in the coming decades. ESSE 21 offers an expanded infrastructure for an interactive community of educators and researchers including minority participants that develops interdisciplinary Earth system science content. Emphasis is on the utilization of NASA resources involving global change data, models, visualizations and electronic media and networks. The ultimate aim of ESSE 21 is to expand and accelerate the nation's realization of sound, scientific interdisciplinary educational resources for informed learning and decision-making by all from the perspective of sustainability of the Earth as a system. The next Call for Participation will be released in late 2003.

JPRS Report, Science & Technology, Japan, MITI’s Large-Scale R&D Projects Reviewed

DTIC Science & Technology

1990-02-08

pollutions, red tide, Active enzymes etc. for cleaners and detergents -- .... .... Intermediates aw materials for r rcosmetics and and medicines moisturizers...PN 00 1H carq 1 0 HZc4 IO -l~ to u .ci~’ *H 0 w 1 Q 0 r - 0 0J w H 04P 04- c0 a) 00 bD O44 0 w w 0 -p 00 021.J4 COQ )a > u0𔃺-0 T1 CL Cfp p P H -H14 OL

ReAsH/FlAsH labeling and image analysis of tetracysteine sensor proteins in cells.

PubMed

Irtegun, Sevgi; Ramdzan, Yasmin M; Mulhern, Terrence D; Hatters, Danny M

2011-08-31

Fluorescent proteins and dyes are essential tools for the study of protein trafficking, localization and function in cells. While fluorescent proteins such as green fluorescence protein (GFP) have been extensively used as fusion partners to proteins to track the properties of a protein of interest, recent developments with smaller tags enable new functionalities of proteins to be examined in cells such as conformational change and protein-association. One small tag system involves a tetracysteine motif (CCXXCC) genetically inserted into a target protein, which binds to biarsenical dyes, ReAsH (red fluorescent) and FlAsH (green fluorescent), with high specificity even in live cells. The TC/biarsenical dye system offers far less steric constraints to the host protein than fluorescent proteins which has enabled several new approaches to measure conformational change and protein-protein interactions. We recently developed a novel application of TC tags as sensors of oligomerization in cells expressing mutant huntingtin, which when mutated aggregates in neurons in Huntington disease. Huntingtin was tagged with two fluorescent dyes, one a fluorescent protein to track protein location, and the second a TC tag which only binds biarsenical dyes in monomers. Hence, changes in colocalization between protein and biarsenical dye reactivity enabled submicroscopic oligomer content to be spatially mapped within cells. Here, we describe how to label TC-tagged proteins fused to a fluorescent protein (Cherry, GFP or CFP) with FlAsH or ReAsH in live mammalian cells and how to quantify the two color fluorescence (Cherry/FlAsH, CFP/FlAsH or GFP/ReAsH combinations).

Response of knee fibrocartilage to joint destabilization.

PubMed

Dyment, N A; Hagiwara, Y; Jiang, X; Huang, J; Adams, D J; Rowe, D W

2015-06-01

A major challenge to understanding osteoarthritis (OA) pathology is identifying the cellular events that precede the onset of cartilage damage. The objective of this study is to determine the effect of joint destabilization on early changes to fibrocartilage in the joint. The anterior cruciate ligament was transected in collagen reporter mice (Col1CFP and ColXRFP). Mineralization labels were given every 2 weeks to measure new mineralized cartilage apposition. Novel fluorescent histology of mineralized tissue was used to characterize the changes in fibrocartilage at 2 and 4 weeks post-injury. Changes in fibrocartilaginous structures of the joint occur as early as 2 weeks after injury and are well developed by 4 weeks. The alterations are seen in multiple entheses and in the medial surface of the femoral and tibial condyles. In the responding entheses, mineral apposition towards the ligament midsubstance results in thickening of the mineralize fibrocartilage. These changes are associated with increases in ColX-RFP, Col1-CFP reporter activity and alkaline phosphatase enzyme activity. Mineral apposition also occurs in the fibrocartilage of the non-articular regions of the medial condyles by 2 weeks and develops into osteophytes by 4 weeks post-injury. An unexpected observation is punctate expression of tartrate resistant acid phosphatase activity in unmineralized fibrochondrocytes adjacent to active appositional mineralization. These observations suggest that fibrocartilage activates prior to degradation of the articular cartilage. Thus clinical and histological imaging of fibrocartilage may be an earlier indicator of disease initiation and may indicate a more appropriate time to start preventative treatment. Copyright © 2015 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Crystal field analysis of the energy level structure of Cs2NaAlF6:Cr3+

NASA Astrophysics Data System (ADS)

Rudowicz, C.; Brik, M. G.; Avram, N. M.; Yeung, Y. Y.; Gnutek, P.

2006-06-01

An analysis of the energy level structure of Cr3+ ions in Cs2NaAlF6 crystal is performed using the exchange charge model (ECM) together with the crystal field analysis/microscopic spin Hamiltonian (CFA/MSH) computer package. Utilizing the crystal structure data, our approach enables modelling of the crystal field parameters (CFPs) and thus the energy level structure for Cr3+ ions at the two crystallographically inequivalent sites in Cs2NaAlF6. Using the ECM initial adjustment procedure, the CFPs are calculated in the crystallographic axis system centred at the Cr3+ ion at each site. Additionally the CFPs are also calculated using the superposition model (SPM). The ECM and SPM predicted CFP values match very well. Consideration of the symmetry aspects for the so-obtained CFP datasets reveals that the latter axis system matches the symmetry-adapted axis system related directly to the six Cr-F bonds well. Using the ECM predicted CFPs as an input for the CFA/MSH package, the complete energy level schemes are calculated for Cr3+ ions at the two sites. Comparison of the theoretical results with the experimental spectroscopic data yields satisfactory agreement. Our results confirm that the actual symmetry at both impurity sites I and II in the Cs2NaAlF6:Cr3+ system is trigonal D3d. The ECM predicted CFPs may be used as the initial (starting) parameters for simulations and fittings of the energy levels for Cr3+ ions in structurally similar hosts.

Periprosthetic bone remodelling of short-stem total hip arthroplasty: a systematic review.

PubMed

Yan, Shuang G; Weber, Patrick; Steinbrück, Arnd; Hua, Xingyi; Jansson, Volkmar; Schmidutz, Florian

2017-11-27

Short-stem hip arthroplasty (SHA) was designed to preserve bone stock and provide an improved load transfer. To gain more evidence regarding the load transfer, this review analysed the periprosthetic bone remodelling of SHA in comparison to standard hip arthroplasty (THA). PubMed and ScienceDirect were screened to extract dual-energy X-ray absorptiometry (DXA) studies evaluating the periprosthetic bone remodelling of SHA and two proven THA designs. From the studies included, the postoperative change in periprosthetic bone mineral density (BMD) after one year and the trend over two years was determined. Fifteen studies with four SHAs (CFP, Metha, Nanos, Fitmore) and two THAs (CLS and Bicontact) designs were included. All SHA and THA stems revealed an initial decrease at the calcar and major trochanter (Gruen 1 and 7) with the Metha, Nanos and Fitmore showing a smaller and more balanced remodelling compared to THA. The pattern after one year and the trend over two years argue for a methaphyseal anchorage of the Metha and Nanos, whereas the Fitmore and CFP seem to anchor metha-diaphyseal. Clearly different pattern of bone remodelling were observed between all four SHAs. Periprosthetic bone remodelling is also present in SHA, with the main bone reduction observed proximally. However, certain SHA stems show a more balanced remodelling compared to THA, arguing for a favourable load transfer. Also, the femoral length where bone remodelling occurs is clearly shorter in SHA. As distinctively different pattern between the SHA designs were observed, they should not be judged as a single implant group.

Assessing flow paths in a karst aquifer based on multiple dye tracing tests using stochastic simulation and the MODFLOW-CFP code

NASA Astrophysics Data System (ADS)

Assari, Amin; Mohammadi, Zargham

2017-09-01

Karst systems show high spatial variability of hydraulic parameters over small distances and this makes their modeling a difficult task with several uncertainties. Interconnections of fractures have a major role on the transport of groundwater, but many of the stochastic methods in use do not have the capability to reproduce these complex structures. A methodology is presented for the quantification of tortuosity using the single normal equation simulation (SNESIM) algorithm and a groundwater flow model. A training image was produced based on the statistical parameters of fractures and then used in the simulation process. The SNESIM algorithm was used to generate 75 realizations of the four classes of fractures in a karst aquifer in Iran. The results from six dye tracing tests were used to assign hydraulic conductivity values to each class of fractures. In the next step, the MODFLOW-CFP and MODPATH codes were consecutively implemented to compute the groundwater flow paths. The 9,000 flow paths obtained from the MODPATH code were further analyzed to calculate the tortuosity factor. Finally, the hydraulic conductivity values calculated from the dye tracing experiments were refined using the actual flow paths of groundwater. The key outcomes of this research are: (1) a methodology for the quantification of tortuosity; (2) hydraulic conductivities, that are incorrectly estimated (biased low) with empirical equations that assume Darcian (laminar) flow with parallel rather than tortuous streamlines; and (3) an understanding of the scale-dependence and non-normal distributions of tortuosity.

3D Cloud Field Prediction using A-Train Data and Machine Learning Techniques

NASA Astrophysics Data System (ADS)

Johnson, C. L.

2017-12-01

Validation of cloud process parameterizations used in global climate models (GCMs) would greatly benefit from observed 3D cloud fields at the size comparable to that of a GCM grid cell. For the highest resolution simulations, surface grid cells are on the order of 100 km by 100 km. CloudSat/CALIPSO data provides 1 km width of detailed vertical cloud fraction profile (CFP) and liquid and ice water content (LWC/IWC). This work utilizes four machine learning algorithms to create nonlinear regressions of CFP, LWC, and IWC data using radiances, surface type and location of measurement as predictors and applies the regression equations to off-track locations generating 3D cloud fields for 100 km by 100 km domains. The CERES-CloudSat-CALIPSO-MODIS (C3M) merged data set for February 2007 is used. Support Vector Machines, Artificial Neural Networks, Gaussian Processes and Decision Trees are trained on 1000 km of continuous C3M data. Accuracy is computed using existing vertical profiles that are excluded from the training data and occur within 100 km of the training data. Accuracy of the four algorithms is compared. Average accuracy for one day of predicted data is 86% for the most successful algorithm. The methodology for training the algorithms, determining valid prediction regions and applying the equations off-track is discussed. Predicted 3D cloud fields are provided as inputs to the Ed4 NASA LaRC Fu-Liou radiative transfer code and resulting TOA radiances compared to observed CERES/MODIS radiances. Differences in computed radiances using predicted profiles and observed radiances are compared.

Src-dependent phosphorylation of caveolin-1 Tyr-14 promotes swelling and release of caveolae

PubMed Central

Zimnicka, Adriana M.; Husain, Yawer S.; Shajahan, Ayesha N.; Sverdlov, Maria; Chaga, Oleg; Chen, Zhenlong; Toth, Peter T.; Klomp, Jennifer; Karginov, Andrei V.; Tiruppathi, Chinnaswamy; Malik, Asrar B.; Minshall, Richard D.

2016-01-01

Caveolin 1 (Cav1) is a required structural component of caveolae, and its phosphorylation by Src is associated with an increase in caveolae-mediated endocytosis. Here we demonstrate, using quantitative live-cell 4D, TIRF, and FRET imaging, that endocytosis and trafficking of caveolae are associated with a Cav1 Tyr-14 phosphorylation-dependent conformational change, which spatially separates, or loosens, Cav1 molecules within the oligomeric caveolar coat. When tracked by TIRF and spinning-disk microscopy, cells expressing phosphomimicking Cav1 (Y14D) mutant formed vesicles that were greater in number and volume than with Y14F-Cav1-GFP. Furthermore, we observed in HEK cells cotransfected with wild-type, Y14D, or Y14F Cav1-CFP and -YFP constructs that FRET efficiency was greater with Y14F pairs than with Y14D, indicating that pY14-Cav1 regulates the spatial organization of Cav1 molecules within the oligomer. In addition, albumin-induced Src activation or direct activation of Src using a rapamycin-inducible Src construct (RapR-Src) led to an increase in monomeric Cav1 in Western blots, as well as a simultaneous increase in vesicle number and decrease in FRET intensity, indicative of a Src-mediated conformational change in CFP/YFP-tagged WT-Cav1 pairs. We conclude that phosphorylation of Cav1 leads to separation or “spreading” of neighboring negatively charged N-terminal phosphotyrosine residues, promoting swelling of caveolae, followed by their release from the plasma membrane. PMID:27170175

Sustainable use of flatfish resources: Addressing the credibility crisis in mixed fisheries management

NASA Astrophysics Data System (ADS)

Rijnsdorp, A. D.; Daan, N.; Dekker, W.; Poos, J. J.; Van Densen, W. L. T.

2007-02-01

Many flatfish species are caught in mixed demersal trawl fisheries and managed by Total Allowable Catch (TAC). Despite decades of fisheries management, several major stocks are severely depleted. Using the Common Fisheries Policy (CFP) as an example, the failure of mixed-fisheries management is analysed by focussing on: the management system; the role of science; the role of managers and politicians; the response of fisheries to management. Failure of the CFP management could be ascribed to: incorrect management advice owing to bias in stock assessments; the tendency of politicians to set the TAC well above the recommended level; and non-compliance of the fisheries with the management regulations. We conclude that TAC management, although apparently successful in some single-species fisheries, inevitably leads to unsustainable exploitation of stocks caught in mixed demersal fisheries as it promotes discarding of over-quota catch and misreporting of catches, thereby corrupting the basis of the scientific advice and increasing the risk of stock collapse. This failure in mixed demersal fisheries has resulted in the loss of credibility of both scientists and managers, and has undermined the support of fishermen for management regulations. An approach is developed to convert the TAC system into a system that controls the total allowable effort (TAE). The approach takes account of the differences in catch efficiency between fleets as well as seasonal changes in the distribution of the target species and can also be applied in the recovery plans for rebuilding specific components of the demersal fish community, such as plaice, cod and hake.

Engineering fluorescent proteins towards ultimate performances: lessons from the newly developed cyan variants

NASA Astrophysics Data System (ADS)

Mérola, Fabienne; Erard, Marie; Fredj, Asma; Pasquier, Hélène

2016-03-01

New fluorescent proteins (FPs) are constantly discovered from natural sources, and submitted to intensive engineering based on random mutagenesis and directed evolution. However, most of these newly developed FPs fail to achieve all the performances required for their bioimaging applications. The design of highly optimised FP-based reporters, simultaneously displaying appropriate colour, multimeric state, chromophore maturation, brightness, photostability and environmental sensitivity will require a better understanding of the structural and dynamic determinants of FP photophysics. The recent development of cyan fluorescent proteins (CFPs) like mCerulean3, mTurquoise2 and Aquamarine brings a different view on these questions, as in this particular case, a step by step evaluation of critical mutations has been performed within a family of spectrally identical and evolutionary close variants. These efforts have led to CFPs with quantum yields close to unity, near single exponential emission decays, high photostability and complete insensitivity to pH, making them ideal choices as energy transfer donors in FRET and FLIM imaging applications. During this process, it was found that a proper amino-acid choice at only two positions (148 and 65) is sufficient to transform the performances of CFPs: with the help of structural and theoretical investigations, we rationalise here how these two positions critically control the CFP photophysics, in the context of FPs derived from the Aequorea victoria species. Today, these results provide a useful toolbox for upgrading the different CFP donors carried by FRET biosensors. They also trace the route towards the de novo design of FP-based optogenetic devices that will be perfectly tailored to dedicated imaging and sensing applications.

The inner side of T cell lipid rafts.

PubMed

Gri, Giorgia; Molon, Barbara; Manes, Santos; Pozzan, Tullio; Viola, Antonella

2004-07-15

A key question in understanding the functional role of lipid rafts is whether lipid microdomains at the plasma membrane outer leaflet are coupled to lipid microdomains at the inner leaflet. By using a cyan-fluorescent protein (CFP) targeted to inner plasma membrane rafts of Jurkat T cells, we found that raft domains at the outer and inner leaflets are physically coupled and that this coupling requires cholesterol. Interestingly, TCR/CD3 cross-linking induces co-capping of the raft bilayer independently of cholesterol or signaling events, indicating that cholesterol-extracting drugs are unable to destroy TCR-lipid rafts interaction.

Mycobacterium tuberculosis effectors interfering host apoptosis signaling.

PubMed

Liu, Minqiang; Li, Wu; Xiang, Xiaohong; Xie, Jianping

2015-07-01

Tuberculosis remains a serious human public health concern. The coevolution between its pathogen Mycobacterium tuberculosis and human host complicated the way to prevent and cure TB. Apoptosis plays subtle role in this interaction. The pathogen endeavors to manipulate the apoptosis via diverse effectors targeting key signaling nodes. In this paper, we summarized the effectors pathogen used to subvert the apoptosis, such as LpqH, ESAT-6/CFP-10, LAMs. The interplay between different forms of cell deaths, such as apoptosis, autophagy, necrosis, is also discussed with a focus on the modes of action of effectors, and implications for better TB control.

Limitations of Using IL-17A and IFN-γ-Induced Protein 10 to Detect Bovine Tuberculosis

PubMed Central

Xin, Ting; Gao, Xintao; Yang, Hongjun; Li, Pingjun; Liang, Qianqian; Hou, Shaohua; Sui, Xiukun; Guo, Xiaoyu; Yuan, Weifeng; Zhu, Hongfei; Ding, Jiabo; Jia, Hong

2018-01-01

Bovine tuberculosis (bTB) is primarily caused by infection with Mycobacterium bovis, which belongs to the Mycobacterium tuberculosis complex. The airborne route is considered the most common for transmission of M. bovis, and more than 15% of cattle with bTB shed the Mycobacterium, which can be detect by nested PCR to amplify mycobacterial mpb70 from a nasal swab from a cow. To screen for cytokines fostering early and accurate detection of bTB, peripheral blood mononuclear cells were isolated from naturally M. bovis-infected, experimentally M. bovis 68002-infected, and uninfected cattle, then these cells were stimulated by PPD-B, CFP-10-ESAT-6 (CE), or phosphate-buffered saline (PBS) for 6 h. The levels of interferon gamma (IFN-γ), IFN-γ-induced protein 10 (IP-10), IL-6, IL-12, IL-17A, and tumor necrosis factor alpha mRNA were measured using real-time PCR. To explore the cytokines associated with different periods of M. bovis infection, cattle were divided into three groups: PCR-positive, PCR-negative, and uninfected using the tuberculin skin test, CFP-10/ESAT-6/TB10.4 protein cocktail-based skin test, IFN-γ release assay (IGRA), CFP-10/ESAT-6 (CE)-based IGRA, and nested PCR. The expression of IP-10, IL-17A, and IFN-γ proteins induced by PPD-B, CE, or PBS was detected by ELISA. The results showed that levels of PPD-B-stimulated IL-17A and IP-10 (mRNA and protein), and CE-induced IP-10 (mRNA and protein) were significantly higher in cattle naturally or experimentally infected with M. bovis than in those that were uninfected. The levels of PPD-B- or CE-induced IL-17A and IP-10 (protein) could be used to differentiate M. bovis-infected calves from uninfected ones for 6 to 30 weeks post-infection, whereas PPD-B- and CE-induced IP-10 and IL-17A mRNA expression could be used to differentiate M. bovis-infected calves from uninfected ones between 6 and 58 weeks post-infection. However, CE-induced IL-17A (protein) was not a reliable indicator of M. bovis infection in cattle that were confirmed positive for infection by nested PCR. Furthermore, the levels of PPD-B- or CE-induced IP-10 and IL-17A protein were lower than IFN-γ in M. bovis-infected cattle. Therefore, IL-17A and IP-10 protein are not suitable biomarkers for bTB. Antigen-induced IP-10 mRNA should be analyzed further for their potential to be used in the diagnosis of bTB. PMID:29560355

Field Application of Serodiagnostics To Identify Elephants with Tuberculosis prior to Case Confirmation by Culture

PubMed Central

Greenwald, Rena; Esfandiari, Javan; Mikota, Susan; Miller, Michele; Moller, Torsten; Vogelnest, Larry; Gairhe, Kamal P.; Robbe-Austerman, Suelee; Gai, Jackie; Waters, W. Ray

2012-01-01

Three serologic methods for antibody detection in elephant tuberculosis (TB), the multiantigen print immunoassay (MAPIA), ElephantTB STAT-PAK kit, and DPP VetTB test, were evaluated using serial serum samples from 14 captive elephants infected with Mycobacterium tuberculosis in 5 countries. In all cases, serological testing was performed prior to the diagnosis of TB by mycobacterial culture of trunk wash or tissue samples collected at necropsy. All elephants produced antibody responses to M. tuberculosis antigens, with 13/14 recognizing ESAT-6 and/or CFP10 proteins. The findings supported the high serodiagnostic test accuracy in detecting infections months to years before M. tuberculosis could be isolated from elephants. The MAPIA and/or DPP VetTB assay demonstrated the potential for monitoring antimycobacterial therapy and predicting TB relapse in treated elephants when continuously used in the posttreatment period. History of exposure to TB and past treatment information should be taken into consideration for proper interpretation of the antibody test results. Data suggest that the more frequent trunk wash culture testing of seropositive elephants may enhance the efficiency of the TB diagnostic algorithm, leading to earlier treatment with improved outcomes. PMID:22695162

ReAsH/FlAsH Labeling and Image Analysis of Tetracysteine Sensor Proteins in Cells

PubMed Central

Irtegun, Sevgi; Ramdzan, Yasmin M.; Mulhern, Terrence D.; Hatters, Danny M.

2011-01-01

Fluorescent proteins and dyes are essential tools for the study of protein trafficking, localization and function in cells. While fluorescent proteins such as green fluorescence protein (GFP) have been extensively used as fusion partners to proteins to track the properties of a protein of interest1, recent developments with smaller tags enable new functionalities of proteins to be examined in cells such as conformational change and protein-association 2, 3. One small tag system involves a tetracysteine motif (CCXXCC) genetically inserted into a target protein, which binds to biarsenical dyes, ReAsH (red fluorescent) and FlAsH (green fluorescent), with high specificity even in live cells 2. The TC/biarsenical dye system offers far less steric constraints to the host protein than fluorescent proteins which has enabled several new approaches to measure conformational change and protein-protein interactions 4-7. We recently developed a novel application of TC tags as sensors of oligomerization in cells expressing mutant huntingtin, which when mutated aggregates in neurons in Huntington disease 7. Huntingtin was tagged with two fluorescent dyes, one a fluorescent protein to track protein location, and the second a TC tag which only binds biarsenical dyes in monomers. Hence, changes in colocalization between protein and biarsenical dye reactivity enabled submicroscopic oligomer content to be spatially mapped within cells. Here, we describe how to label TC-tagged proteins fused to a fluorescent protein (Cherry, GFP or CFP) with FlAsH or ReAsH in live mammalian cells and how to quantify the two color fluorescence (Cherry/FlAsH, CFP/FlAsH or GFP/ReAsH combinations). PMID:21897361

Development of ball surface acoustic wave trace moisture analyzer using burst waveform undersampling circuit

NASA Astrophysics Data System (ADS)

Tsuji, Toshihiro; Oizumi, Toru; Fukushi, Hideyuki; Takeda, Nobuo; Akao, Shingo; Tsukahara, Yusuke; Yamanaka, Kazushi

2018-05-01

The measurement and control of trace moisture, where the water concentration is lower than 1 ppmv [-76.2 °C for the frost point (°CFP)], are essential for improving the yield rate of semiconductor devices and for ensuring their reliability. A ball surface acoustic wave (SAW) sensor with a sol-gel silica coating exhibited useful characteristics for a trace moisture analyzer (TMA) when the temperature drift of the delay time output was precisely compensated using two-frequency measurement (TFM), where the temperature-compensated relative delay time change (RDTC) was obtained by subtracting the RDTC at the fundamental frequency from that at the third harmonic frequency on an identical propagation path. However, the cost of the measurement circuit was a problem. In this study, a burst waveform undersampling (BUS) circuit based on the theory of undersampling measurement was developed as a practical means. The BUS circuit was useful for precise temperature compensation of the RDTC, and the ball SAW TMA was prototyped by calibrating the RDTC using a TMA based on cavity ring-down spectroscopy (CRDS), which is the most reliable method for trace moisture measurement. The ball SAW TMA outputted a similar concentration to that obtained by the CRDS TMA, and its response time at a set concentration in N2 with a flow rate of 1 l/min was about half that of the CRDS TMA, suggesting that moisture of -80 °CFP was measured within only 1 min. The detection limit at a signal-to-noise ratio of 3 was estimated to be 0.05 ppbv, comparable with that of the CRDS TMA. From these results, it was demonstrated that a practical ball SAW TMA can be realized using the developed BUS circuit.

c-Fos induction in mesotelencephalic dopamine pathway projection targets and dorsal striatum following oral intake of sugars and fats in rats.

PubMed

Dela Cruz, J A D; Coke, T; Karagiorgis, T; Sampson, C; Icaza-Cukali, D; Kest, K; Ranaldi, R; Bodnar, R J

2015-02-01

Overconsumption of nutrients high in fats and sugars can lead to obesity. Previous studies indicate that sugar or fat consumption activate individual brain sites using Fos-like immunoreactivity (FLI). Sugars and fats also elicit conditioned flavor preferences (CFP) that are differentially mediated by flavor-flavor (orosensory: f/f) and flavor-nutrient (post-ingestive: f/n) processes. Dopamine (DA) signaling in the medial prefrontal cortex (mPFC), the amygdala (AMY) and the nucleus accumbens (NAc), has been implicated in acquisition and expression of fat- and sugar-CFP. The present study examined the effects of acute consumption of fat (corn oil: f/f and f/n), glucose (f/f and f/n), fructose, (f/f only), saccharin, xanthan gum or water upon simultaneous FLI activation of DA mesotelencephalic nuclei (ventral tegmental area (VTA)) and projections (infralimbic and prelimbic mPFC, basolateral and central-cortico-medial AMY, core and shell of NAc as well as the dorsal striatum). Consumption of corn oil solutions, isocaloric to glucose and fructose, significantly increased FLI in all sites except for the NAc shell. Glucose intake significantly increased FLI in both AMY areas, dorsal striatum and NAc core, but not in either mPFC area, VTA or Nac shell. Correspondingly, fructose intake significantly increased FLI in the both AMY areas, the infralimbic mPFC and dorsal striatum, but not the prelimbic mPFC, VTA or either NAc area. Saccharin and xanthan gum intake failed to activate FLI relative to water. When significant FLI activation occurred, highly positive relationships were observed among sites, supporting the idea of activation of a distributed brain network mediating sugar and fat intake. Copyright © 2014 Elsevier Inc. All rights reserved.

Immuno Nanosensor for the Ultrasensitive Naked Eye Detection of Tuberculosis.

PubMed

Mohd Bakhori, Noremylia; Yusof, Nor Azah; Abdullah, Jaafar; Wasoh, Helmi; Md Noor, Siti Suraiya; Ahmad Raston, Nurul Hanun; Mohammad, Faruq

2018-06-14

In the present study, a beneficial approach for the ultrasensitive and affordable naked eye detection and diagnosis of tuberculosis (TB) by utilizing plasmonic enzyme-linked immunosorbent assay (ELISA) via antibody-antigen interaction was studied. Here, the biocatalytic cycle of the intracellular enzymes links to the formation and successive growth of the gold nanoparticles (GNPs) for ultrasensitive detection. The formation of different colored solutions by the plasmonic nanoparticles in the presence of enzyme labels links directly to the existence or non-existence of the TB analytes in the sample solutions. For disease detection, the adapted protocol is based mainly on the conventional ELISA procedure that involves catalase-labeled antibodies, i.e., the enzymes consume hydrogen peroxide and further produce GNPs with the addition of gold (III) chloride. The amount of hydrogen peroxide remaining in the solution determines whether the GNPs solution is to be formed in the color blue or the color red, as it serves as a confirmation for the naked eye detection of TB analytes. However, the conventional ELISA method only shows tonal colors that need a high concentration of analyte to achieve high confidence levels for naked eye detection. Also, in this research, we proposed the incorporation of protein biomarker, Mycobacterium tuberculosis ESAT-6-like protein esxB (CFP-10), as a means of TB detection using plasmonic ELISA. With the use of this technique, the CFP-10 detection limit can be lowered to 0.01 µg/mL by the naked eye. Further, our developed technique was successfully tested and confirmed with sputum samples from patients diagnosed with positive TB, thereby providing enough evidence for the utilization of our technique in the early diagnosis of TB disease.

Dihydrolipoamide dehydrogenase-Lpd (Rv0462)-specific T cell recall responses are higher in healthy household contacts of TB: a novel immunodominant antigen from M. tuberculosis.

PubMed

Devasundaram, Santhi; Raja, Alamelu

2017-07-01

The partial effectiveness against pulmonary tuberculosis (PTB), displayed by the existing tuberculosis (TB) vaccine, bacillus Calmette-Guérin (BCG), highlights the need for novel vaccines to replace or improve BCG. In TB immunology, antigen-specific cellular immune response is frequently considered indispensable. Latency-associated antigens are intriguing as targets for TB vaccine development. The mycobacterial protein, dihydrolipoamide dehydrogenase (Lpd; Rv0462), the third enzyme of the pyruvate dehydrogenase (PDH) complex, facilitates Mycobacterium tuberculosis to resist host reactive nitrogen intermediates. Multicolor flow cytometry analysis of whole-blood cultures showed higher Lpd-specific Th1 recall response (IFN-γ, TNF-α, and IL-2; P = 0.0006) and memory CD4 + and CD8 + T cells (CCR7 + CD45RA - and CCR7 - CD45RA - ) in healthy household contacts (HHC) of TB ( P < 0.0001), which is comparable with or higher than the standard antigens, ESAT-6 and CFP-10. The frequency of Lpd-specific multifunctional T cells was higher in HHC compared with PTB patients. However, there is no significant statistical correlation. Regulatory T cell (T reg ) analysis of HHCs and active TB patients demonstrated very low Lpd-specific CD4 + T regs relative to ESAT-6 and CFP-10. Our study demonstrates that the Lpd antigen induces a strong cellular immune response in healthy mycobacteria-infected individuals. In consideration of this population having demonstrated immunologic protection against active TB disease development, our data are encouraging about the possible use of Lpd as a target for further TB subunit vaccine development. © Society for Leukocyte Biology.

Targeting cysteine-mediated dimerization of the MUC1-C oncoprotein in human cancer cells

PubMed Central

RAINA, DEEPAK; AHMAD, REHAN; RAJABI, HASAN; PANCHAMOORTHY, GOVIND; KHARBANDA, SURENDER; KUFE, DONALD

2012-01-01

The MUC1 heterodimeric protein is aberrantly overexpressed in diverse human carcinomas and contributes to the malignant phenotype. The MUC1-C transmembrane subunit contains a CQC motif in the cytoplasmic domain that has been implicated in the formation of dimers and in its oncogenic function. The present study demonstrates that MUC1-C forms dimers in human breast and lung cancer cells. MUC1-C dimerization was detectable in the cytoplasm and was independent of MUC1-N, the N-terminal mucin subunit that extends outside the cell. We show that the MUC1-C cytoplasmic domain forms dimers in vitro that are disrupted by reducing agents. Moreover, dimerization of the MUC1-C subunit in cancer cells was blocked by reducing agents and increased by oxidative stress, supporting involvement of the CQC motif in forming disulfide bonds. In support of these observations, mutation of the MUC1-C CQC motif to AQA completely blocked MUC1-C dimerization. Importantly, this study was performed with MUC1-C devoid of fluorescent proteins, such as GFP, CFP and YFP. In this regard, we show that GFP, CFP and YFP themselves form dimers that are readily detectable with cross-linking agents. The present results further demonstrate that a cell-penetrating peptide that targets the MUC1-C CQC cysteines blocks MUC1-C dimerization in cancer cells. These findings provide definitive evidence that: i) the MUC1-C cytoplasmic domain cysteines are necessary and sufficient for MUC1-C dimerization, and ii) these CQC motif cysteines represent an Achilles’ heel for targeting MUC1-C function. PMID:22200620

Connectivity, excitability and activity patterns in neuronal networks

NASA Astrophysics Data System (ADS)

le Feber, Joost; Stoyanova, Irina I.; Chiappalone, Michela

2014-06-01

Extremely synchronized firing patterns such as those observed in brain diseases like epilepsy may result from excessive network excitability. Although network excitability is closely related to (excitatory) connectivity, a direct measure for network excitability remains unavailable. Several methods currently exist for estimating network connectivity, most of which are related to cross-correlation. An example is the conditional firing probability (CFP) analysis which calculates the pairwise probability (CFPi,j) that electrode j records an action potential at time t = τ, given that electrode i recorded a spike at t = 0. However, electrode i often records multiple spikes within the analysis interval, and CFP values are biased by the on-going dynamic state of the network. Here we show that in a linear approximation this bias may be removed by deconvoluting CFPi,j with the autocorrelation of i (i.e. CFPi,i), to obtain the single pulse response (SPRi,j)—the average response at electrode j to a single spike at electrode i. Thus, in a linear system SPRs would be independent of the dynamic network state. Nonlinear components of synaptic transmission, such as facilitation and short term depression, will however still affect SPRs. Therefore SPRs provide a clean measure of network excitability. We used carbachol and ghrelin to moderately activate cultured cortical networks to affect their dynamic state. Both neuromodulators transformed the bursting firing patterns of the isolated networks into more dispersed firing. We show that the influence of the dynamic state on SPRs is much smaller than the effect on CFPs, but not zero. The remaining difference reflects the alteration in network excitability. We conclude that SPRs are less contaminated by the dynamic network state and that mild excitation may decrease network excitability, possibly through short term synaptic depression.

Reforming Biomass Derived Pyrolysis Bio-oil Aqueous Phase to Fuels

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

Mukarakate, Calvin; Evans, Robert J.; Deutch, Steve

Fast pyrolysis and catalytic fast pyrolysis (CFP) of biomass produce a liquid product stream comprised of various classes of organic compounds having different molecule size and polarity. This liquid, either spontaneously in the case of catalytic fast pyrolysis or by water addition for the non-catalytic process separates into a non-polar organic-rich fraction and a highly polar water-rich fraction. The organic fraction can be used as a blendstock or feedstock for further processing in a refinery while, in the CFP process design, the aqueous phase is currently sent to wastewater treatment, which results in a loss of residual biogenic carbon presentmore » in this stream. Our work focuses on the catalytic conversion of the biogenic carbon in pyrolysis aqueous phase streams to produce hydrocarbons using a vertical micro-reactor coupled to a molecular beam mass spectrometer (MBMS). Furthermore, the MBMS provides real-time analysis of products while also tracking catalyst deactivation. The catalyst used in this work was HZSM-5, which upgraded the oxygenated organics in the aqueous fraction to fuels comprising small olefins and aromatic hydrocarbons. During processing the aqueous bio-oil fraction the HZSM-5 catalyst exhibited higher activity and coke resistance than those observed in similar experiments using biomass or whole bio-oils. Reduced coking is likely due to ejection of coke precursors from the catalyst pores that was enhanced by excess process water available for steam stripping. The water reacted with coke precursors to form phenol, methylated phenols, naphthol, and methylated naphthols. Conversion data shows that up to 40 wt% of the carbon in the feed stream is recovered as hydrocarbons.« less

Comparative Effectiveness of Traditional Chinese Medicine (TCM) and Psychosocial Care in the Treatment of TMD-associated Chronic Facial Pain

PubMed Central

Ritenbaugh, Cheryl; Hammerschlag, Richard; Dworkin, Samuel F; Aickin, Mikel G; Mist, Scott D; Elder, Charles; Harris, Richard E

2012-01-01

Summary This dual-site study sought to identify the appropriate role for TCM (acupuncture and herbs) in conjunction with a validated psychosocial self-care intervention (SC) for treating chronic TMD-associated pain. Participants with RDC-TMD-confirmed TMD (n=168) entered a stepped-care protocol that began with a basic TMD class. At weeks 2 and 10, patients receiving SC whose worst facial pain was above predetermined levels were reallocated by minimization to SC or TCM with experienced practitioners. Characteristic facial pain (CFP: mean of worst pain, average pain when having pain, current pain; each VAS 0-10) was the primary outcome. Social activity interference (VAS 0-10) was a secondary outcome. Patients were monitored for safety. TCM provided significantly greater short-term (8-week) relief than SC (CFP reduction difference, −0.60 [SDE 0.26], p=0.020), and greater reduction in interference with social activities (−0.81 [SDE 0.33], p=0.016). In two of five treatment trajectory groups, more than 2/3 of participants demonstrated clinically meaningful responses (> 30% improvement) in pain interference over 16 weeks. This study provides evidence that TMD patients referred for TCM in a community-based model will receive safe treatment that is likely to provide some short-term pain relief and improved quality of life. Similar designs may also apply to evaluations of other kinds of chronic pain. (ClinicalTrials.gov number NCT00856167) PERSPECTIVE This short-term comparative effectiveness study of chronic facial pain suggests that Traditional Chinese Medicine is safe and frequently efficacious alone or subsequent to standard psychosocial interventions. TCM is widely available throughout North America and may provide clinicians and patients with a reasonable addition or alternative to other forms of therapy. PMID:23059454

Fluorescence-based characterization of genetically encoded peptides that fold in live cells: progress toward a generic hairpin scaffold

NASA Astrophysics Data System (ADS)

Cheng, Zihao; Campbell, Robert E.

2007-02-01

Binding proteins suitable for expression and high affinity molecular recognition in the cytoplasm or nucleus of live cells have numerous applications in the biological sciences. In an effort to add a new minimal motif to the growing repertoire of validated non-immunoglobulin binding proteins, we have undertaken the development of a generic protein scaffold based on a single β-hairpin that can fold efficiently in the cytoplasm. We have developed a method, based on the measurement of fluorescence resonance energy transfer (FRET) between a genetically fused cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP), that allows the structural stability of recombinant β-hairpin peptides to be rapidly assessed both in vitro and in vivo. We have previously reported the validation of this method when applied to a 16mer tryptophan zipper β-hairpin. We now describe the use of this method to evaluate the potential of a designed 20mer β-hairpin peptide with a 3rd Trp/Trp cross-strand pair to function as a generic protein scaffold. Quantitative analysis of the FRET efficiency, resistance to proteolysis (assayed by loss of FRET), and circular dichroism spectra revealed that the 20mer peptide is significantly more tolerant of destabilizing mutations than the 16mer peptide. Furthermore, we experimentally demonstrate that the in vitro determined β-hairpin stabilities are well correlated with in vivo β-hairpin stabilities as determined by FRET measurements of colonies of live bacteria expressing the recombinant peptides flanked by CFP and YFP. Finally, we report on our progress to develop highly folded 24mer and 28mer β-hairpin peptides through the use of fluorescence-based library screening.

The C2 domain of PKCalpha is a Ca2+ -dependent PtdIns(4,5)P2 sensing domain: a new insight into an old pathway.

PubMed

Sánchez-Bautista, Sonia; Marín-Vicente, Consuelo; Gómez-Fernández, Juan C; Corbalán-García, Senena

2006-10-06

The C2 domain is a targeting domain that responds to intracellular Ca2+ signals in classical protein kinases (PKCs) and mediates the translocation of its host protein to membranes. Recent studies have revealed a new motif in the C2 domain, named the lysine-rich cluster, that interacts with acidic phospholipids. The purpose of this work was to characterize the molecular mechanism by which PtdIns(4,5)P2 specifically interacts with this motif. Using a combination of isothermal titration calorimetry, fluorescence resonance energy transfer and time-lapse confocal microscopy, we show here that Ca2+ specifically binds to the Ca2+ -binding region, facilitating PtdIns(4,5)P2 access to the lysine-rich cluster. The magnitude of PtdIns(4,5)P2 binding is greater than in the case of other polyphosphate phosphatidylinositols. Very importantly, the residues involved in PtdIns(4,5)P2 binding are essential for the plasma membrane localization of PKCalpha when RBL-2H3 cells are stimulated through their IgE receptors. Additionally, CFP-PH and CFP-C1 domains were used as bioprobes to demonstrate the co-existence of PtdIns(4,5)P2 and diacylglycerol in the plasma membrane, and it was shown that although a fraction of PtdIns(4,5)P2 is hydrolyzed to generate diacylglycerol and IP3, an important amount still remains in the membrane where it is available to activate PKCalpha. These findings entail revision of the currently accepted model of PKCalpha recruitment to the membrane and its activation.

Reforming Biomass Derived Pyrolysis Bio-oil Aqueous Phase to Fuels

DOE PAGES

Mukarakate, Calvin; Evans, Robert J.; Deutch, Steve; ...

2017-01-07

Fast pyrolysis and catalytic fast pyrolysis (CFP) of biomass produce a liquid product stream comprised of various classes of organic compounds having different molecule size and polarity. This liquid, either spontaneously in the case of catalytic fast pyrolysis or by water addition for the non-catalytic process separates into a non-polar organic-rich fraction and a highly polar water-rich fraction. The organic fraction can be used as a blendstock or feedstock for further processing in a refinery while, in the CFP process design, the aqueous phase is currently sent to wastewater treatment, which results in a loss of residual biogenic carbon presentmore » in this stream. Our work focuses on the catalytic conversion of the biogenic carbon in pyrolysis aqueous phase streams to produce hydrocarbons using a vertical micro-reactor coupled to a molecular beam mass spectrometer (MBMS). Furthermore, the MBMS provides real-time analysis of products while also tracking catalyst deactivation. The catalyst used in this work was HZSM-5, which upgraded the oxygenated organics in the aqueous fraction to fuels comprising small olefins and aromatic hydrocarbons. During processing the aqueous bio-oil fraction the HZSM-5 catalyst exhibited higher activity and coke resistance than those observed in similar experiments using biomass or whole bio-oils. Reduced coking is likely due to ejection of coke precursors from the catalyst pores that was enhanced by excess process water available for steam stripping. The water reacted with coke precursors to form phenol, methylated phenols, naphthol, and methylated naphthols. Conversion data shows that up to 40 wt% of the carbon in the feed stream is recovered as hydrocarbons.« less

Influence of pesticide regulation on acute poisoning deaths in Sri Lanka.

PubMed Central

Roberts, Darren M.; Karunarathna, Ayanthi; Buckley, Nick A.; Manuweera, Gamini; Sheriff, M. H. Rezvi; Eddleston, Michael

2003-01-01

OBJECTIVES: To assess in a developing Asian country the impact of pesticide regulation on the number of deaths from poisoning. These regulations, which were implemented in Sri Lanka from the 1970s, aimed to reduce the number of deaths - the majority from self-poisoning - by limiting the availability and use of highly toxic pesticides. METHODS: Information on legislative changes was obtained from the Ministry of Agriculture, national and district hospital admission data were obtained from the Sri Lanka Health Statistics Unit, and individual details of deaths by pesticide poisoning were obtained from a manual review of patients' notes and intensive care unit records in Anuradhapura. FINDINGS: Between 1986 and 2000, the total national number of admissions due to poisoning doubled, and admissions due to pesticide poisoning increased by more than 50%. At the same time, the case fatality proportion (CFP) fell for total poisonings and for poisonings due to pesticides. In 1991_92, 72% of pesticide-induced deaths in Anuradhapura were caused by organophosphorus (OP) and carbamate pesticides - in particular, the WHO class I OPs monocrotophos and methamidophos. From 1991, the import of these pesticides was reduced gradually until they were banned for routine use in January 1995, with a corresponding fall in deaths. Unfortunately, their place in agricultural practice was taken by the WHO class II organochlorine endosulfan, which led to a rise in deaths from status epilepticus - from one in 1994 to 50 in 1998. Endosulfan was banned in 1998, and over the following three years the number of endosulfan deaths fell to three. However, at the end of the decade, the number of deaths from pesticides was at a similar level to that of 1991, with WHO class II OPs causing the most deaths. Although these drugs are less toxic than class I OPs, the management of class II OPs remains difficult because they are, nevertheless, still highly toxic, and their toxicity is exacerbated by the paucity of available facilities. CONCLUSION: The fall in CFP amidst a rising incidence of self-poisoning suggests that Sri Lanka's programmes of pesticide regulation were beneficial. However, a closer inspection of pesticide-induced deaths in one hospital revealed switching to other highly toxic pesticides, as one was banned and replaced in agricultural practice by another. Future regulation must predict this switching and bear in mind the ease of treatment of replacement pesticides. Furthermore, such regulations must be implemented alongside other strategies, such as integrated pest management, to reduce the overall pesticide availability for self-harm. PMID:14758405

TROTT Computer Program for Two-Dimensional Stress Wave Propagation

DTIC Science & Technology

1980-04-01

1 ,4 3. OOOE- 08 3.000E+08 6, .2 17 7,1 D00E+09 NSIZE 0 0 0 8 8 8 0 0 0 YIELD = 1 .030E+10 8, 190E+11 PBXN - 106 RHOS = 1 .634 CFP = i ooo DPY = i...TROTT 104 TROTT 105 TROTT 106 TROTT 107 TROTT 106 TROTT 109 TROTT 110 TROTT 11 1 TROTT 112 TROTT 113 TROTT 114 TROTT 1 15 TROTT 1 16...TROTT 158 TROTT 159 TROTT 160 TROTT 161 TROTT 162 TROTT 163 TROTT 164 TROTT 165 TROTT 166 TROTT 167 TROTT 168 TROTT 169 TROTT 170 106

Production of antibodies against glycolipids from the Mycobacterium tuberculosis cell wall in aerosol murine models of tuberculosis.

PubMed

Cardona, P J; Julián, E; Vallès, X; Gordillo, S; Muñoz, M; Luquin, M; Ausina, V

2002-06-01

Evolution of antibodies against glycolipids from the Mycobacterium tuberculosis cell wall has been studied for the first time in experimental murine models of tuberculosis induced by aerosol, in which infection, reinfection, reactivation, prophylaxis and treatment with antibiotics have been assayed. Results show a significant humoral response against these antigens, where diacyltrehaloses (DAT) and sulpholipid I (SL-I) elicited higher antibody levels than protein antigens like antigen 85 protein complex (Ag85), culture filtrate proteins (CFP) and purified protein derivative (PPD). Only immunoglobulin M (IgM) antibodies have been detected against DAT and SL-I. Their evolution has a positive correlation with bacillary concentration in tissues.

Visualization of the activation of the histamine H3 receptor (H3R) using novel fluorescence resonance energy transfer biosensors and their potential application to the study of H3R pharmacology.

PubMed

Liu, Ying; Zeng, Hong; Pediani, John D; Ward, Richard J; Chen, Lu-Yao; Wu, Nan; Ma, Li; Tang, Mei; Yang, Yang; An, Su; Guo, Xiao-Xi; Hao, Qian; Xu, Tian-Rui

2018-06-01

Activation of the histamine-3 receptor (H3R) is involved in memory processes and cognitive action, while blocking H3R activation can slow the progression of neurological disorders, such as Alzheimer's disease, schizophrenia and narcolepsy. To date, however, no direct way to examine the activation of H3R has been utilized. Here, we describe a novel biosensor that can visualize the activation of H3R through an intramolecular fluorescence resonance energy transfer (FRET) signal. To achieve this, we constructed an intramolecular H3R FRET sensor with cyan fluorescent protein (CFP) attached at the C terminus and yellow fluorescent protein (YFP) inserted into the third intracellular loop. The sensor was found to internalize normally on agonist treatment. We measured FRET signals between the donor CFP and the acceptor YFP in living cells in real time, the results of which indicated that H3R agonist treatment (imetit or histamine) increases the FRET signal in a time- and concentration-dependent manner with Kon and Koff values consistent with published data and which maybe correlated with decreasing cAMP levels and the promotion of ERK1/2 phosphorylation. The FRET signal was inhibited by H3R antagonists, and the introduction of mutations at F419A, F423A, L426A and L427A, once again, the promotion of ERK1/2 phosphorylation, was diminished. Thus, we have built a H3R biosensor which can visualize the activation of receptor through real-time structure changes and which can obtain pharmacological kinetic data at the same time. The FRET signals may allow the sensor to become a useful tool for screening compounds and optimizing useful ligands. © 2018 Federation of European Biochemical Societies.

Cross reactive immune responses in cattle arising from exposure to Mycobacterium bovis and non-tuberculous mycobacteria.

PubMed

Jenkins, A O; Gormley, E; Gcebe, N; Fosgate, G T; Conan, A; Aagaard, C; Michel, A L; Rutten, V P M G

2018-04-01

Accurate diagnosis of tuberculosis in cattle may be compromised in areas where there are high rates of exposure to environmental/non-tuberculous mycobacteria (NTM). This cross reaction of immune responses to Mycobacterium bovis antigens shared with NTMs can result in reduced specificity of commonly used diagnostic tests including tuberculin skin tests and the interferon gamma assay (IFN-ɣ). In this study we assessed the cross-reactive immune responses of M. bovis (infected) and NTM exposed animals to M. bovis and M. avium tuberculin, the ESAT6/CFP10 cocktail antigen, tuberculin derived from cultures of selected NTMs, and a panel of recombinant mycobacterium tuberculosis complex (MTBC) antigens sharing homology with orthologues in NTM. Gamma interferon (IFN-ɣ) responses were measured in whole blood cultures using the IFN-ɣ assay and the IFN-ɣ elispot assay on purified peripheral blood mononuclear cells (PBMC). We observed the expected strong IFN-ɣ response to PPD-B in the M. bovis infected animals that distinguished this group from non-infected NTM exposed cattle. The IFN-ɣ responses to PPD-N (M. nonchromogenicum), were relatively high in both infected and non-infected NTM exposed cattle, but were not significantly different to classify the true infection status of each group. The results indicated that the cross-reactive responses to PPD-B and/or PPD-A with PPD-N, likely arose from prior exposure to environmental non-tuberculous mycobacteria. The IFN-ɣ immune responses to the 10 R-Mag measured by the IFN-ɣ elispot assay revealed that three of the selected antigens, Rv3615 (ESpC), Rv0287 (esxG) and the ESAT6/CFP10, were immunogenic in the infected cattle, and distinguished the infected cattle from the non-infected NTM exposed animals. The combined data of PPDs and R-Mags derived from NTM mycobacteria may prove useful in future development of novel bTB diagnostic tests. Copyright © 2018 Elsevier B.V. All rights reserved.

Improved method for efficient imaging of intracellular Cl− with Cl-Sensor using conventional fluorescence setup

PubMed Central

Friedel, Perrine; Bregestovski, Piotr; Medina, Igor

2013-01-01

Chloride (Cl−) homeostasis is known to be fundamental for central nervous system functioning. Alterations in intracellular Cl− concentration ([Cl−]i) and changes in the efficacy of Cl− extrusion are involved in numerous neurological disorders. Therefore, there is a strong need for studies of the dynamics of [Cl−]i in different cell types under physiological conditions and during pathology. Several previous works reported having successfully achieved recording of [Cl−]i using genetically encoded Cl-Sensor that is composed of the cyan fluorescent protein (CFP) and Cl−-sensitive mutant of the yellow fluorescent protein (YFPCl). However, all reported works were performed using specially designed setups with ultra-sensitive CCD cameras. Our multiple attempts to monitor Cl−-dependent fluorescence of Cl-Sensor using conventional epifluorescence microscopes did not yield successful results. In the present work, we have analysed the reason of our failures and found that they were caused by a strong inactivation of the YFPCl component of Cl-Sensor during excitation of the CFP with 430 nm light. Based on the obtained results, we reduced 20-fold the intensity of the 430 nm excitation and modified the recording protocol that allows now stable long-lasting ratiometric measurements of Cl-Sensor fluorescence in different cell types including cultured hippocampal neurons and their tiny dendrites and spines. Simultaneous imaging and patch clamp recording revealed that in mature neurons, the novel protocol allows detection of as little as 2 mM changes of [Cl−]i from the resting level of 5–10 mM. We demonstrate also a usefulness of the developed [Cl−]i measurement procedure for large scale screening of the activity of exogenously expressed potassium-chloride co-transporter KCC2, a major neuronal Cl− extruder that is implicated in numerous neurological disorders and is a target for novel therapeutical treatments. PMID:23596389

Thermodynamic Stability of Molybdenum Oxycarbides Formed from Orthorhombic Mo 2 C in Oxygen-Rich Environments

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

Likith, S. R. J.; Farberow, C. A.; Manna, S.

Molybdenum carbide (Mo 2C) nanoparticles and thin films are particularly suitable catalysts for catalytic fast pyrolysis (CFP) as they are effective for deoxygenation and can catalyze certain reactions that typically occur on noble metals. Oxygen deposited during deoxygenation reactions may alter the carbide structure, leading to the formation of oxycarbides, which can determine changes in catalytic activity or selectivity. Despite emerging spectroscopic evidence of bulk oxycarbides, so far there have been no reports of their precise atomic structure or their relative stability with respect to orthorhombic Mo 2C. This knowledge is essential for assessing the catalytic properties of molybdenum (oxy)carbidesmore » for CFP. In this article, we use density functional theory (DFT) calculations to (a) describe the thermodynamic stability of surface and subsurface configurations of oxygen and carbon atoms for a commonly studied Mo-terminated surface of orthorhombic Mo 2C and (b) determine atomic structures for oxycarbides with a Mo:C ratio of 2:1. The surface calculations suggest that oxygen atoms are not stable under the top Mo layer of the Mo 2C(100) surface. Coupling DFT calculations with a polymorph sampling method, we determine (Mo 2C) xO y oxycarbide structures for a wide range of oxygen compositions. Oxycarbides with lower oxygen content (y/x = 2) adopt layered structures reminiscent of the parent carbide phase, with flat Mo layers separated by layers of oxygen and carbon; for higher oxygen content, our results suggest the formation of amorphous phases, as the atomic layers lose their planarity with increasing oxygen content. We characterize the oxidation states of Mo in the oxycarbide structures determined computationally, and simulate their X-ray diffraction (XRD) patterns in order to facilitate comparisons with experiments. Our study may provide a platform for large-scale investigations of the catalytic properties of oxycarbides and their surfaces and for tailoring the catalytic properties for different desired reactions.« less

The ΔfbpA attenuated candidate vaccine from Mycobacterium tuberculosis, H37Rv primes for a stronger T-bet dependent Th1 immunity in mice.

PubMed

Roche, Cherie M; Smith, Amanda; Lindsey, Devin R; Meher, Akshay; Schluns, Kimberly; Arora, Ashish; Armitige, Lisa Y; Jagannath, Chinnaswamy

2011-12-01

The ΔfbpA candidate vaccine derived from Mycobacterium tuberculosis (H37Rv) (Mtb) protects mice better than BCG against tuberculosis, and we investigated the hypothesis that ΔfbpA may induce a stronger Th1 immunity. Since T-bet transcription factor regulates Th1 immunity, mice infected with ΔfbpA, BCG vaccine and related mycobacteria were analyzed for T-bet positive T cells. Mouse dendritic cells (DCs) or macrophages were also pulsed with excretory-secreted antigens (ES; Antigen-85B, ESAT-6 and CFP10) and cocultured with T cells from immunized or naïve mice and tested for in vitro induction of T-bet and IFN-γ. In both models, ΔfbpA mutant induced a stronger response of T-bet(+)CD4 T cells, which correlated with an increased expansion of IFN-γ(+)CD4 T cells in vivo and in vitro. When DCs pulsed with ES antigens were allowed to stimulate T cells, ESAT-6 and CFP-10 failed to induce a recall expansion of T-bet(+)IFN-γ(+)CD4 T cells from BCG vaccinated mice. Thus, deletion of RD1 in BCG seems to reduce its ability to induce T-bet and induce stronger Th1 immunity. Finally, mice were vaccinated with ΔfbpA and BCG and challenged with virulent Mtb for evaluation of protection and T cell expansion. ΔfbpA vaccinated mice showed a rapid and stronger expansion of CD4(+)CXCR3(+) IFN-γ(+) T cells in the lungs of Mtb challenged mice, compared to those which had BCG vaccine. ΔfbpA immunized mice also showed a better decline of the Mtb bacterial counts of the lungs. Mtb derived ΔfbpA candidate vaccine therefore induces qualitatively better T-bet dependent Th1 immunity than BCG vaccine. Copyright © 2011 Elsevier Ltd. All rights reserved.

Improved method for efficient imaging of intracellular Cl(-) with Cl-Sensor using conventional fluorescence setup.

PubMed

Friedel, Perrine; Bregestovski, Piotr; Medina, Igor

2013-01-01

Chloride (Cl(-)) homeostasis is known to be fundamental for central nervous system functioning. Alterations in intracellular Cl(-) concentration ([Cl(-)]i) and changes in the efficacy of Cl(-) extrusion are involved in numerous neurological disorders. Therefore, there is a strong need for studies of the dynamics of [Cl(-)]i in different cell types under physiological conditions and during pathology. Several previous works reported having successfully achieved recording of [Cl(-)]i using genetically encoded Cl-Sensor that is composed of the cyan fluorescent protein (CFP) and Cl(-)-sensitive mutant of the yellow fluorescent protein (YFPCl). However, all reported works were performed using specially designed setups with ultra-sensitive CCD cameras. Our multiple attempts to monitor Cl(-)-dependent fluorescence of Cl-Sensor using conventional epifluorescence microscopes did not yield successful results. In the present work, we have analysed the reason of our failures and found that they were caused by a strong inactivation of the YFPCl component of Cl-Sensor during excitation of the CFP with 430 nm light. Based on the obtained results, we reduced 20-fold the intensity of the 430 nm excitation and modified the recording protocol that allows now stable long-lasting ratiometric measurements of Cl-Sensor fluorescence in different cell types including cultured hippocampal neurons and their tiny dendrites and spines. Simultaneous imaging and patch clamp recording revealed that in mature neurons, the novel protocol allows detection of as little as 2 mM changes of [Cl(-)]i from the resting level of 5-10 mM. We demonstrate also a usefulness of the developed [Cl(-)]i measurement procedure for large scale screening of the activity of exogenously expressed potassium-chloride co-transporter KCC2, a major neuronal Cl(-) extruder that is implicated in numerous neurological disorders and is a target for novel therapeutical treatments.

β-Subunits Promote the Expression of CaV2.2 Channels by Reducing Their Proteasomal Degradation*

PubMed Central

Waithe, Dominic; Ferron, Laurent; Page, Karen M.; Chaggar, Kanchan; Dolphin, Annette C.

2011-01-01

The β-subunits of voltage-gated calcium channels regulate their functional expression and properties. Two mechanisms have been proposed for this, an effect on gating and an enhancement of expression. With respect to the effect on expression, β-subunits have been suggested to enhance trafficking by masking an unidentified endoplasmic reticulum (ER) retention signal. Here we have investigated whether, and how, β-subunits affect the level of CaV2.2 channels within somata and neurites of cultured sympathetic neurons. We have used YFP-CaV2.2 containing a mutation (W391A), that prevents binding of β-subunits to its I-II linker and found that expression of this channel was much reduced compared with WT CFP-CaV2.2 when both were expressed in the same neuron. This effect was particularly evident in neurites and growth cones. The difference between the levels of YFP-CaV2.2(W391A) and CFP-CaV2.2(WT) was lost in the absence of co-expressed β-subunits. Furthermore, the relative reduction of expression of CaV2.2(W391A) compared with the WT channel was reversed by exposure to two proteasome inhibitors, MG132 and lactacystin, particularly in the somata. In further experiments in tsA-201 cells, we found that proteasome inhibition did not augment the cell surface CaV2.2(W391A) level but resulted in the observation of increased ubiquitination, particularly of mutant channels. In contrast, we found no evidence for selective retention of CaV2.2(W391A) in the ER, in either the soma or growth cones. In conclusion, there is a marked effect of β-subunits on CaV2.2 expression, particularly in neurites, but our results point to protection from proteasomal degradation rather than masking of an ER retention signal. PMID:21233207

Hierarchy Low CD4+/CD8+ T-Cell Counts and IFN-γ Responses in HIV-1+ Individuals Correlate with Active TB and/or M.tb Co-Infection.

PubMed

Shao, Lingyun; Zhang, Xinyun; Gao, Yan; Xu, Yunya; Zhang, Shu; Yu, Shenglei; Weng, Xinhua; Shen, Hongbo; Chen, Zheng W; Jiang, Weimin; Zhang, Wenhong

2016-01-01

Detailed studies of correlation between HIV-M.tb co-infection and hierarchy declines of CD8+/CD4+ T-cell counts and IFN-γ responses have not been done. We conducted case-control studies to address this issue. 164 HIV-1-infected individuals comprised of HIV-1+ATB, HIV-1+LTB and HIV-1+TB- groups were evaluated. Immune phenotyping and complete blood count (CBC) were employed to measure CD4+ and CD8+ T-cell counts; T.SPOT.TB and intracellular cytokine staining (ICS) were utilized to detect ESAT6, CFP10 or PPD-specific IFN-γ responses. There were significant differences in median CD4+ T-cell counts between HIV-1+ATB (164/μL), HIV-1+LTB (447/μL) and HIV-1+TB- (329/μL) groups. Hierarchy low CD4+ T-cell counts (<200/μL, 200-500/μL, >500/μL) were correlated significantly with active TB but not M.tb co-infection. Interestingly, hierarchy low CD8+ T-cell counts were not only associated significantly with active TB but also with M.tb co-infection (P<0.001). Immunologically, HIV-1+ATB group showed significantly lower numbers of ESAT-6-/CFP-10-specific IFN-γ+ T cells than HIV-1+LTB group. Consistently, PPD-specific IFN-γ+CD4+/CD8+ T effector cells in HIV-1+ATB group were significantly lower than those in HIV-1+LTB group (P<0.001). Hierarchy low CD8+ T-cell counts and effector function in HIV-1-infected individuals are correlated with both M.tb co-infection and active TB. Hierarchy low CD4+ T-cell counts and Th1 effector function in HIV-1+ individuals are associated with increased frequencies of active TB, but not M.tb co-infection.

The prevalence of benthic dinoflagellates associated with ciguatera fish poisoning in the central Red Sea.

PubMed

Catania, Daniela; Richlen, Mindy L; Mak, Yim Ling; Morton, Steve L; Laban, Elizabeth H; Xu, Yixiao; Anderson, Donald M; Chan, Leo Lai; Berumen, Michael L

2017-09-01

This study confirms the presence of the toxigenic benthic dinoflagellates Gambierdiscus belizeanus and Ostreopsis spp. in the central Red Sea. To our knowledge, this is also the first report of these taxa in coastal waters of Saudi Arabia, indicating the potential occurrence of ciguatera fish poisoning (CFP) in that region. During field investigations carried out in 2012 and 2013, a total of 100 Turbinaria and Halimeda macroalgae samples were collected from coral reefs off the Saudi Arabian coast and examined for the presence of Gambierdiscus and Ostreopsis, two toxigenic dinoflagellate genera commonly observed in coral reef communities around the world. Both Gambierdiscus and Ostreopsis spp. were observed at low densities (<200 cells g -1 wet weight algae). Cell densities of Ostreopsis spp. were significantly higher than Gambierdiscus spp. at most of the sampling sites, and abundances of both genera were negatively correlated with seawater salinity. To assess the potential for ciguatoxicity in this region, several Gambierdiscus isolates were established in culture and examined for species identity and toxicity. All isolates were morphologically and molecularly identified as Gambierdiscus belizeanus. Toxicity analysis of two isolates using the mouse neuroblastoma cell-based assay for ciguatoxins (CTX) confirmed G. belizeanus as a CTX producer, with a maximum toxin content of 6.50±1.14×10 -5 pg P-CTX-1 eq. cell -1 . Compared to Gambierdiscus isolates from other locations, these were low toxicity strains. The low Gambierdiscus densities observed along with their comparatively low toxin contents may explain why CFP is unidentified and unreported in this region. Nevertheless, the presence of these potentially toxigenic dinoflagellate species at multiple sites in the central Red Sea warrants future study on their possible effects on marine food webs and human health in this region. Copyright © 2017 Elsevier B.V. All rights reserved.

Thermodynamic Stability of Molybdenum Oxycarbides Formed from Orthorhombic Mo 2 C in Oxygen-Rich Environments

DOE PAGES

Likith, S. R. J.; Farberow, C. A.; Manna, S.; ...

2017-12-20

Molybdenum carbide (Mo 2C) nanoparticles and thin films are particularly suitable catalysts for catalytic fast pyrolysis (CFP) as they are effective for deoxygenation and can catalyze certain reactions that typically occur on noble metals. Oxygen deposited during deoxygenation reactions may alter the carbide structure, leading to the formation of oxycarbides, which can determine changes in catalytic activity or selectivity. Despite emerging spectroscopic evidence of bulk oxycarbides, so far there have been no reports of their precise atomic structure or their relative stability with respect to orthorhombic Mo 2C. This knowledge is essential for assessing the catalytic properties of molybdenum (oxy)carbidesmore » for CFP. In this article, we use density functional theory (DFT) calculations to (a) describe the thermodynamic stability of surface and subsurface configurations of oxygen and carbon atoms for a commonly studied Mo-terminated surface of orthorhombic Mo 2C and (b) determine atomic structures for oxycarbides with a Mo:C ratio of 2:1. The surface calculations suggest that oxygen atoms are not stable under the top Mo layer of the Mo 2C(100) surface. Coupling DFT calculations with a polymorph sampling method, we determine (Mo 2C) xO y oxycarbide structures for a wide range of oxygen compositions. Oxycarbides with lower oxygen content (y/x = 2) adopt layered structures reminiscent of the parent carbide phase, with flat Mo layers separated by layers of oxygen and carbon; for higher oxygen content, our results suggest the formation of amorphous phases, as the atomic layers lose their planarity with increasing oxygen content. We characterize the oxidation states of Mo in the oxycarbide structures determined computationally, and simulate their X-ray diffraction (XRD) patterns in order to facilitate comparisons with experiments. Our study may provide a platform for large-scale investigations of the catalytic properties of oxycarbides and their surfaces and for tailoring the catalytic properties for different desired reactions.« less

Dual-Mode SERS-Fluorescence Immunoassay Using Graphene Quantum Dot Labeling on One-Dimensional Aligned Magnetoplasmonic Nanoparticles.

PubMed

Zou, Fengming; Zhou, Hongjian; Tan, Tran Van; Kim, Jeonghyo; Koh, Kwangnak; Lee, Jaebeom

2015-06-10

A novel dual-mode immunoassay based on surface-enhanced Raman scattering (SERS) and fluorescence was designed using graphene quantum dot (GQD) labels to detect a tuberculosis (TB) antigen, CFP-10, via a newly developed sensing platform of linearly aligned magnetoplasmonic (MagPlas) nanoparticles (NPs). The GQDs were excellent bilabeling materials for simultaneous Raman scattering and photoluminescence (PL). The one-dimensional (1D) alignment of MagPlas NPs simplified the immunoassay process and enabled fast, enhanced signal transduction. With a sandwich-type immunoassay using dual-mode nanoprobes, both SERS signals and fluorescence images were recognized in a highly sensitive and selective manner with a detection limit of 0.0511 pg mL(-1).

A C. trachomatis Cloning Vector and the Generation of C. trachomatis Strains Expressing Fluorescent Proteins under the Control of a C. trachomatis Promoter

PubMed Central

Agaisse, Hervé; Derré, Isabelle

2013-01-01

Here we describe a versatile cloning vector for conducting genetic experiments in C. trachomatis. We successfully expressed various fluorescent proteins (i.e. GFP, mCherry and CFP) from C. trachomatis regulatory elements (i.e. the promoter and terminator of the incDEFG operon) and showed that the transformed strains produced wild type amounts of infectious particles and recapitulated major features of the C. trachomatis developmental cycle. C. trachomatis strains expressing fluorescent proteins are valuable tools for studying the C. trachomatis developmental cycle. For instance, we show the feasibility of investigating the dynamics of inclusion fusion and interaction with host proteins and organelles by time-lapse video microscopy. PMID:23441233

Fluorescent Protein-Based Ca2+ Sensor Reveals Global, Divalent Cation-Dependent Conformational Changes in Cardiac Troponin C.

PubMed

Badr, Myriam A; Pinto, Jose R; Davidson, Michael W; Chase, P Bryant

2016-01-01

Cardiac troponin C (cTnC) is a key effector in cardiac muscle excitation-contraction coupling as the Ca2+ sensing subunit responsible for controlling contraction. In this study, we generated several FRET sensors for divalent cations based on cTnC flanked by a donor fluorescent protein (CFP) and an acceptor fluorescent protein (YFP). The sensors report Ca2+ and Mg2+ binding, and relay global structural information about the structural relationship between cTnC's N- and C-domains. The sensors were first characterized using end point titrations to decipher the response to Ca2+ binding in the presence or absence of Mg2+. The sensor that exhibited the largest responses in end point titrations, CTV-TnC, (Cerulean, TnC, and Venus) was characterized more extensively. Most of the divalent cation-dependent FRET signal originates from the high affinity C-terminal EF hands. CTV-TnC reconstitutes into skinned fiber preparations indicating proper assembly of troponin complex, with only ~0.2 pCa unit rightward shift of Ca2+-sensitive force development compared to WT-cTnC. Affinity of CTV-TnC for divalent cations is in agreement with known values for WT-cTnC. Analytical ultracentrifugation indicates that CTV-TnC undergoes compaction as divalent cations bind. C-terminal sites induce ion-specific (Ca2+ versus Mg2+) conformational changes in cTnC. Our data also provide support for the presence of additional, non-EF-hand sites on cTnC for Mg2+ binding. In conclusion, we successfully generated a novel FRET-Ca2+ sensor based on full length cTnC with a variety of cellular applications. Our sensor reveals global structural information about cTnC upon divalent cation binding.

Synthesis of ultrathin mesoporous NiCo2O4 nanosheets on carbon fiber paper as integrated high-performance electrodes for supercapacitors

NASA Astrophysics Data System (ADS)

Deng, Fangze; Yu, Lin; Cheng, Gao; Lin, Ting; Sun, Ming; Ye, Fei; Li, Yongfeng

2014-04-01

Two-dimensional ultrathin mesoporous NiCo2O4 nanosheets on carbon fiber paper (CFP) are synthesized through a facile solvothermal method combined with a post thermal treatment. The well interconnected ultrathin NiCo2O4 nanosheets directly grown on the carbon nanofibers could allow for easy diffusion of the electrolyte, shorten the transport path of ion and electron and accommodate the strain during cycling. As a result, superior pseudocapacitive performance is achieved with large specific capacitance of 999 F g-1 at a high current density of 20 A g-1. The capacitance loss is 15.6% after 3000 cycles at a current density of 10 A g-1, displaying good cycle ability and high rate capability.

Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells

PubMed Central

Warren, Sean C.; Margineanu, Anca; Katan, Matilda; Dunsby, Chris; French, Paul M. W.

2015-01-01

Multiplexed imaging of Förster Resonance Energy Transfer (FRET)-based biosensors potentially presents a powerful approach to monitoring the spatio-temporal correlation of signalling pathways within a single live cell. Here, we discuss the potential of homo-FRET based biosensors to facilitate multiplexed imaging. We demonstrate that the homo-FRET between pleckstrin homology domains of Akt (Akt-PH) labelled with mCherry may be used to monitor 3′-phosphoinositide accumulation in live cells and show how global analysis of time resolved fluorescence anisotropy measurements can be used to quantify this accumulation. We further present multiplexed imaging readouts of calcium concentration, using fluorescence lifetime measurements of TN-L15-a CFP/YFP based hetero-FRET calcium biosensor-with 3′-phosphoinositide accumulation. PMID:26133241

Inhibition of α-glucosidase by polysaccharides from the fruit hull of Camellia oleifera Abel.

PubMed

Zhang, Sheng; Li, Xiang-Zhou

2015-01-22

We isolated and purified polysaccharides from the Camellia oleifera Abel. fruit hull and studied its hypoglycemic potential. Our results revealed six polysaccharides (CFPA-1-5 & CFPB) from the aqueous extract from the defatted C. oleifera fruit hull. Purified polysaccharides (purity >90%) were investigated for the inhibition of α-glucosidase activity in vitro. Two polysaccharides, CFPB and CFPA-3 were present in high concentration in the fruit hull and showed a dose-dependent inhibition of α-glucosidase activity, with IC50 concentrations of 11.80 and 10.95 μg/mL, respectively. This result suggests that polysaccharides (CFP) extracted from the fruit hull of C. oleifera may have potential as functional foods with featuring a hypoglycemic effect. Copyright © 2014 Elsevier Ltd. All rights reserved.

Meta-analyses of the sensitivity and specificity of ante-mortem and post-mortem diagnostic tests for bovine tuberculosis in the UK and Ireland.

PubMed

Nuñez-Garcia, Javier; Downs, Sara H; Parry, Jessica E; Abernethy, Darrell A; Broughan, Jennifer M; Cameron, Angus R; Cook, Alasdair J; de la Rua-Domenech, Ricardo; Goodchild, Anthony V; Gunn, Jane; More, Simon J; Rhodes, Shelley; Rolfe, Simon; Sharp, Michael; Upton, Paul A; Vordermeier, H Martin; Watson, Eamon; Welsh, Michael; Whelan, Adam O; Woolliams, John A; Clifton-Hadley, Richard S; Greiner, Matthias

2018-05-01

Bovine Tuberculosis (bTB) in cattle is a global health problem and eradication of the disease requires accurate estimates of diagnostic test performance to optimize their efficiency. The objective of this study was, through statistical meta-analyses, to obtain estimates of sensitivity (Se) and specificity (Sp), for 14 different ante-mortem and post-mortem diagnostic tests for bTB in cattle. Using data from a systematic review of the scientific literature (published 1934-2009) diagnostic Se and Sp were estimated using Bayesian logistic regression models adjusting for confounding factors. Random effect terms were used to account for unexplained heterogeneity. Parameters in the models were implemented using Markov Chain Monte Carlo (MCMC), and posterior distributions for the diagnostic parameters with adjustment for covariates (confounding factors) were obtained using the inverse logit function. Estimates for Se and/or Sp of the tuberculin skin tests and the IFN-γ blood test were compared with estimates published 2010-2015. Median Se for the single intradermal comparative cervical tuberculin skin (SICCT) test (standard interpretation) was 0.50 and Bayesian credible intervals (CrI) were wide (95% CrI 0.26, 0.78). Median Sp for the SICCT test was 1.00 (95% CrI 0.99, 1.00). Estimates for the IFN-γ blood test Bovine Purified Protein Derivative (PPD)-Avian PPD and Early Secreted Antigen target 6 and Culture Filtrate Protein 10 (ESAT-6/CFP10) ESAT6/CFP10 were 0.67 (95% CrI 0.49, 0.82) and 0.78 (95% CrI 0.60, 0.90) respectively for Se, and 0.98 (95% CrI 0.96, 0.99) and 0.99 (95% CrI 0.99, 1.00) for Sp. The study provides an overview of the accuracy of a range of contemporary diagnostic tests for bTB in cattle. Better understanding of diagnostic test performance is essential for the design of effective control strategies and their evaluation. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

Detection of relatively penicillin G-resistant Neisseria meningitidis by disk susceptibility testing.

PubMed Central

Campos, J; Mendelman, P M; Sako, M U; Chaffin, D O; Smith, A L; Sáez-Nieto, J A

1987-01-01

Beginning in 1985, relatively penicillin G-resistant (Penr) meningococci which did not produce beta-lactamase were isolated from the blood and cerebrospinal fluid of patients in Spain. We identified 16 Penr (mean MIC, 0.3 microgram/ml; range, 0.1 to 0.7 microgram/ml) and 12 penicillin-susceptible (Pens; mean MIC, less than or equal to 0.06 microgram/ml) strains of Neisseria meningitidis by the agar dilution technique using an inoculum of 10(4) CFU and questioned which disk susceptibility test would best differentiate these two populations. We compared the disk susceptibility of these strains using disks containing 2 (P2) and 10 (P10) U of penicillin G, 2 (Am2) and 10 (Am10) micrograms of ampicillin, and 1 microgram of oxacillin (OX1). We also investigated susceptibility with disks containing 30 micrograms of each of cephalothin (CF30), cefoxitin (FOX30), cefuroxime (CXM30), and cefotaxime (CTX30) and 75 micrograms of cefoperazone (CFP75) and determined by cluster analysis any correlation with the zone diameters obtained with P2 disks. Using the P2 and AM2 disks (in contrast to the P10 and AM10 disks), we correctly differentiated all the Penr from Pens isolates. In addition, the zone diameters with the P2 disk gave the best correlation with the penicillin G MIC determinations. All 16 Penr strains and 3 of 12 Pens strains showed zone diameters of 6 mm around OX1 disks, limiting the usefulness of OX1 disks. The zone diameters obtained with CF30, CXM30, and OX1 disks correlated with those obtained with the P2 disk, which suggests that these antibiotics have similar effects on these strains. In contrast, the data obtained with FOX30, CTX30, and CFP75 disks did not cluster with those obtained with the P2 disk, which suggests that there was a difference in the bacterial target or reflects their greater activity. We conclude that the P2 disk tests more readily identify Penr meningococci than do the standard P10 disk tests. PMID:3124729

Functional Fluorescent Protein Insertions in Herpes Simplex Virus gB Report on gB Conformation before and after Execution of Membrane Fusion

PubMed Central

Gallagher, John R.; Atanasiu, Doina; Saw, Wan Ting; Paradisgarten, Matthew J.; Whitbeck, J. Charles; Eisenberg, Roselyn J.; Cohen, Gary H.

2014-01-01

Entry of herpes simplex virus (HSV) into a target cell requires complex interactions and conformational changes by viral glycoproteins gD, gH/gL, and gB. During viral entry, gB transitions from a prefusion to a postfusion conformation, driving fusion of the viral envelope with the host cell membrane. While the structure of postfusion gB is known, the prefusion conformation of gB remains elusive. As the prefusion conformation of gB is a critical target for neutralizing antibodies, we set out to describe its structure by making genetic insertions of fluorescent proteins (FP) throughout the gB ectodomain. We created gB constructs with FP insertions in each of the three globular domains of gB. Among 21 FP insertion constructs, we found 8 that allowed gB to remain membrane fusion competent. Due to the size of an FP, regions in gB that tolerate FP insertion must be solvent exposed. Two FP insertion mutants were cell-surface expressed but non-functional, while FP insertions located in the crown were not surface expressed. This is the first report of placing a fluorescent protein insertion within a structural domain of a functional viral fusion protein, and our results are consistent with a model of prefusion HSV gB constructed from the prefusion VSV G crystal structure. Additionally, we found that functional FP insertions from two different structural domains could be combined to create a functional form of gB labeled with both CFP and YFP. FRET was measured with this construct, and we found that when co-expressed with gH/gL, the FRET signal from gB was significantly different from the construct containing CFP alone, as well as gB found in syncytia, indicating that this construct and others of similar design are likely to be powerful tools to monitor the conformation of gB in any model system accessible to light microscopy. PMID:25233449

Difference in trafficking of brain-derived neurotrophic factor between axons and dendrites of cortical neurons, revealed by live-cell imaging

PubMed Central

Adachi, Naoki; Kohara, Keigo; Tsumoto, Tadaharu

2005-01-01

Background Brain-derived neurotrophic factor (BDNF), which is sorted into a regulated secretory pathway of neurons, is supposed to act retrogradely through dendrites on presynaptic neurons or anterogradely through axons on postsynaptic neurons. Depending on which is the case, the pattern and direction of trafficking of BDNF in dendrites and axons are expected to be different. To address this issue, we analyzed movements of green fluorescent protein (GFP)-tagged BDNF in axons and dendrites of living cortical neurons by time-lapse imaging. In part of the experiments, the expression of BDNF tagged with cyan fluorescent protein (CFP) was compared with that of nerve growth factor (NGF) tagged with yellow fluorescent protein (YFP), to see whether fluorescent protein-tagged BDNF is expressed in a manner specific to this neurotrophin. Results We found that BDNF tagged with GFP or CFP was expressed in a punctated manner in dendrites and axons in about two-thirds of neurons into which plasmid cDNAs had been injected, while NGF tagged with GFP or YFP was diffusely expressed even in dendrites in about 70% of the plasmid-injected neurons. In neurons in which BDNF-GFP was expressed as vesicular puncta in axons, 59 and 23% of the puncta were moving rapidly in the anterograde and retrograde directions, respectively. On the other hand, 64% of BDNF-GFP puncta in dendrites did not move at all or fluttered back and forth within a short distance. The rest of the puncta in dendrites were moving relatively smoothly in either direction, but their mean velocity of transport, 0.47 ± 0.23 (SD) μm/s, was slower than that of the moving puncta in axons (0.73 ± 0.26 μm/s). Conclusion The present results show that the pattern and velocity of the trafficking of fluorescence protein-tagged BDNF are different between axons and dendrites, and suggest that the anterograde transport in axons may be the dominant stream of BDNF to release sites. PMID:15969745

Genetic variation and effects of candidate-gene polymorphisms on coagulation properties, curd firmness modeling and acidity in milk from Brown Swiss cows.

PubMed

Cecchinato, A; Chessa, S; Ribeca, C; Cipolat-Gotet, C; Bobbo, T; Casellas, J; Bittante, G

2015-07-01

The aims of this study were to estimate the genetic variation of traditional milk coagulation properties (MCPs), milk acidity, curd firmness (CF) modeled on time t (CF(t) ; comprising: RCT(eq), rennet coagulation time estimated from the equation; CF(P), the asymptotic potential curd firmness; k(CF), the curd firming instant rate constant; and k(SR), the syneresis instant rate constant) and maximum CF traits (MCF; comprising CF(max), the maximum CF value; and tmax, the time of attainment). Furthermore, we investigated 96 single nucleotide polymorphisms (SNPs) from 54 candidate genes, testing their associations with the above-listed traits. Milk and blood samples were collected from 1271 cows (each sampled once) from 85 herds. Genotyping was performed using a custom Illumina VeraCode GoldenGate approach. A Bayesian linear animal model (including the effects of herd, days in milk, parity and additive polygenic effects) was used to estimate the genetic parameters of the studied traits. The same model with the addition of the SNP genotype effect was used for our association analysis. The heritability estimates of CF t and the MCF traits (RCT(eq)=0.258; k(CF)=0.230; CF(max)=0.191; t(max)=0.278) were similar to those obtained using traditional MCPs (0.187 to 0.267), except for the lower estimates for CF(P) (0.064) and k(SR) (0.077). A total of 13 of the 51 tested SNPs had relevant additive effects on at least one trait. We observed associations between MCPs and SNPs in the genes encoding ATP-binding cassette sub-family G member 2 (ABCG2), chemokine ligand 2 (CCL2), growth hormone 1 (GH1), prolactin (PRL) and toll-like receptor 2 (TLR2). Whereas, CF(t) and the MCF traits were associated with polymorphisms in the α-s1-casein (CSN1S1), β-casein (CSN2), GH1, oxidized low-density lipoprotein receptor 1 (OLR1), phospholipase C β1 (PLCB1), PRL and signal transducer and activator of transcription 5A (STAT5A) genes.

Crystal-field energy level analysis for Nd(3+) ions at the low symmetry C(1) site in [Nd(hfa)(4)(H(2)O)](N(C(2)H(5))(4)) single crystals.

PubMed

Mech, Agnieszka; Gajek, Zbigniew; Karbowiak, Mirosław; Rudowicz, Czesław

2008-09-24

Optical absorption measurements of Nd(3+) ions in single crystals of [Nd(hfa)(4)(H(2)O)](N(C(2)H(5))(4)) (hfa = hexafluoroacetyloacetonate), denoted Nd(hfa) for short, have been carried out at 4.2 and 298 K. This compound crystallizes in the monoclinic system (space group P 2(1)/n). Each Nd ion is coordinated to eight oxygen atoms that originate from the hexafluoroacetylacetonate ligands and one oxygen atom from the water molecule. A total of 85 experimental crystal-field (CF) energy levels arising from the Nd(3+) (4f(3)) electronic configuration were identified in the optical spectra and assigned. A three-step CF analysis was carried out in terms of a parametric Hamiltonian for the actual C(1) symmetry at the Nd(3+) ion sites. In the first step, a total of 27 CF parameters (CFPs) in the Wybourne notation B(kq), admissible by group theory, were determined in a preliminary fitting constrained by the angular overlap model predictions. The resulting CFP set was reduced to 24 specific independent CFPs using appropriate standardization transformations. Optimizations of the second-rank CFPs and extended scanning of the parameter space were employed in the second step to improve reliability of the CFP sets, which is rather a difficult task in the case of no site symmetry. Finally, seven free-ion parameters and 24 CFPs were freely varied, yielding an rms deviation between the calculated energy levels and the 85 observed ones of 11.1 cm(-1). Our approach also allows prediction of the energy levels of Nd(3+) ions that are hidden in the spectral range overlapping with strong ligand absorption, which is essential for understanding the inter-ionic energy transfer. The orientation of the axis system associated with the fitted CF parameters w.r.t. the crystallographic axes is established. The procedure adopted in our calculations may be considered as a general framework for analysis of CF levels of lanthanide ions at low (triclinic) symmetry sites.

Crystal-field energy level analysis for Nd3+ ions at the low symmetry C1 site in [Nd(hfa)4(H2O)](N(C2H5)4) single crystals

NASA Astrophysics Data System (ADS)

Mech, Agnieszka; Gajek, Zbigniew; Karbowiak, Mirosław; Rudowicz, Czesław

2008-09-01

Optical absorption measurements of Nd3+ ions in single crystals of [Nd(hfa)4(H2O)](N(C2H5)4) (hfa = hexafluoroacetyloacetonate), denoted Nd(hfa) for short, have been carried out at 4.2 and 298 K. This compound crystallizes in the monoclinic system (space group P 21/n). Each Nd ion is coordinated to eight oxygen atoms that originate from the hexafluoroacetylacetonate ligands and one oxygen atom from the water molecule. A total of 85 experimental crystal-field (CF) energy levels arising from the Nd3+ (4f3) electronic configuration were identified in the optical spectra and assigned. A three-step CF analysis was carried out in terms of a parametric Hamiltonian for the actual C1 symmetry at the Nd3+ ion sites. In the first step, a total of 27 CF parameters (CFPs) in the Wybourne notation Bkq, admissible by group theory, were determined in a preliminary fitting constrained by the angular overlap model predictions. The resulting CFP set was reduced to 24 specific independent CFPs using appropriate standardization transformations. Optimizations of the second-rank CFPs and extended scanning of the parameter space were employed in the second step to improve reliability of the CFP sets, which is rather a difficult task in the case of no site symmetry. Finally, seven free-ion parameters and 24 CFPs were freely varied, yielding an rms deviation between the calculated energy levels and the 85 observed ones of 11.1 cm-1. Our approach also allows prediction of the energy levels of Nd3+ ions that are hidden in the spectral range overlapping with strong ligand absorption, which is essential for understanding the inter-ionic energy transfer. The orientation of the axis system associated with the fitted CF parameters w.r.t. the crystallographic axes is established. The procedure adopted in our calculations may be considered as a general framework for analysis of CF levels of lanthanide ions at low (triclinic) symmetry sites.

Invasive Lionfish (Pterosis volitans) Pose Public Health Threats.

PubMed

Diaz, James H

2015-01-01

The lionfish, Pterosis volitans, a native of Indo-Pacific oceans, is a popular saltwater aquarium fish despite venomous spines on its fins. Lionfish were inadvertently introduced into the western Atlantic from Florida in the early 1990s and have overpopulated and dispersed widely into the Caribbean Sea and Gulf of Mexico. Initiatives to control lionfish populations were launched, including the National Oceanographic and Atmospheric Administration (NOAA)-sponsored "Lionfish as Food Campaign".2 Recently, scientists from the Food and Drug Administration (FDA) reported that lionfish caught off the US Virgin Islands contained ciguatoxins and could cause ciguatera fish poisoning (CFP); a seafood-borne poisoning without an antidote or any specific treatment, and a potential for prolonged neurotoxicity. Lionfish pose several public health threats. New strategies to control the lionfish population explosion in coastal waters and offshore fisheries are needed now to ensure seafood safety and public health. The lionfish, Pterosis volitans, is native to the reefs of the western Indian and Pacific Oceans (Figure 1). Brightly colored with red, white, and black stripes and adorned with feathery fins, the lionfish is a popular saltwater aquarium fish despite venomous spines on its fins (Figure 2). Lionfish were introduced into the western North Atlantic from Florida in the early 1990s after some specimens were discarded by dissatisfied amateur aquarists and others escaped from hurricane-flooded public aquariums.1 Since lionfish are voracious carnivores, have few natural predators, and reproduce prolifically, they have overpopulated and dispersed widely from Cape Hatteras to Florida, throughout the Caribbean Sea, and into the Gulf of Mexico.1 The population density of lionfish in its new, invaded territory now exceeds that of its native habitat.1 As a result, campaigns to control lionfish populations were launched in Florida and the Caribbean. Lionfish now pose several public health threats that include (1) serving as the second most common cause of venomous fish puncture injuries next to stingrays; (2) interrupting the marine seafood chain on reef systems that support commercial fisheries; and (3) bioconcentrating heat-stable algal toxins capable of causing CFP.

Non-Target Screening of Veterinary Drugs Using Tandem Mass Spectrometry on SmartMass

NASA Astrophysics Data System (ADS)

Xia, Bing; Liu, Xin; Gu, Yu-Cheng; Zhang, Zhao-Hui; Wang, Hai-Yan; Ding, Li-Sheng; Zhou, Yan

2013-05-01

Non-target screening of veterinary drugs using tandem mass spectrometric data was performed on the SmartMass platform. This newly developed software uses the characteristic fragmentation patterns (CFP) to identify chemicals, especially those containing particular substructures. A mixture of 17 sulfonamides was separated by ultra performance liquid chromatography (UPLC), and SmartMass was used to process the tandem mass spectrometry (MS/MS) data acquired on an Orbitrap mass spectrometer. The data were automatically extracted, and each sulfonamide was recognized and analyzed with a prebuilt analysis rule. By using this software, over 98 % of the false candidate structures were eliminated, and all the correct structures were found within the top 10 of the ranking lists. Furthermore, SmartMass could also be used to identify slightly modified contraband drugs and metabolites with simple prebuilt rules. [Figure not available: see fulltext.

Field Investigation and Modeling Development for Hydrological and Carbon Cycles in Southwest Karst Region of China

NASA Astrophysics Data System (ADS)

Hu, X. B.

2017-12-01

It is required to understanding water cycle and carbon cycle processes for water resource management and pollution prevention and global warming influence in southwest karst region of China. Lijiang river basin is selected as our study region. Interdisciplinary field and laboratory experiments with various technologies are conducted to characterize the karst aquifers in detail. Key processes in the karst water cycle and carbon cycle are determined. Based on the MODFLOW-CFP model, new watershed flow and carbon cycle models are developed coupled subsurface and surface water flow models. Our study focus on the karst springshed in Mao village, the mechanisms coupling carbon cycle and water cycle are explored. This study provides basic theory and simulation method for water resource management and groundwater pollution prevention in China karst region.

Correlative Förster Resonance Electron Transfer-Proximity Ligation Assay (FRET-PLA) Technique for Studying Interactions Involving Membrane Proteins.

PubMed

Ivanusic, Daniel; Denner, Joachim; Bannert, Norbert

2016-08-01

This unit provides a guide and detailed protocol for studying membrane protein-protein interactions (PPI) using the acceptor-sensitized Förster resonance electron transfer (FRET) method in combination with the proximity ligation assay (PLA). The protocol in this unit is focused on the preparation of FRET-PLA samples and the detection of correlative FRET/PLA signals as well as on the analysis of FRET-PLA data and interpretation of correlative results when using cyan fluorescent protein (CFP) as a FRET donor and yellow fluorescent protein (YFP) as a FRET acceptor. The correlative application of FRET and PLA combines two powerful tools for monitoring PPI, yielding results that are more reliable than with either technique alone. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

Application of the FRET method for monitoring the dynamics of caspase-3 activation during apoptosis in living cells

NASA Astrophysics Data System (ADS)

Chen, Tongsheng; Xing, Da

2005-01-01

Activation of caspase-3 is a central event in apoptosis. A fluorescence techniques, fluorescence resonance energy transfer (FRET), was used to study the dynamic of caspase-3 activation during apoptosis induced by tumor necrosis factor TNF-α in living cells. The FRET probe consists a CFP (cyan fluorescent protein) and a Venus (YFP mutant, yellow fluorescent protein) with a specialized linker containing the caspase-3 cleavage sequence: DEVD (Luo et al., 2001). Human lung adenocarcinoma cell line (ASTC-a-1) were stably expressed with the FRET probe and then were treated by TNF-α, respectively. Experimental results showed that FRET could monitor more insensitively the dynamic of caspase-3 activation in real-time in vivo, and this technique will be highly useful for correlating the caspase-3 activation with other apoptotic events and for rapid-screening of potential drugs that may target the apoptotic process.

Redox sensor proteins for highly sensitive direct imaging of intracellular redox state.

PubMed

Sugiura, Kazunori; Nagai, Takeharu; Nakano, Masahiro; Ichinose, Hiroshi; Nakabayashi, Takakazu; Ohta, Nobuhiro; Hisabori, Toru

2015-02-13

Intracellular redox state is a critical factor for fundamental cellular functions, including regulation of the activities of various metabolic enzymes as well as ROS production and elimination. Genetically-encoded fluorescent redox sensors, such as roGFP (Hanson, G. T., et al. (2004)) and Redoxfluor (Yano, T., et al. (2010)), have been developed to investigate the redox state of living cells. However, these sensors are not useful in cells that contain, for example, other colored pigments. We therefore intended to obtain simpler redox sensor proteins, and have developed oxidation-sensitive fluorescent proteins called Oba-Q (oxidation balance sensed quenching) proteins. Our sensor proteins derived from CFP and Sirius can be used to monitor the intracellular redox state as their fluorescence is drastically quenched upon oxidation. These blue-shifted spectra of the Oba-Q proteins enable us to monitor various redox states in conjunction with other sensor proteins. Copyright © 2015 Elsevier Inc. All rights reserved.

An in vitro FRET-based assay for the analysis of SUMO conjugation and isopeptidase cleavage.

PubMed

Stankovic-Valentin, Nicolas; Kozaczkiewicz, Lukasz; Curth, Katja; Melchior, Frauke

2009-01-01

To measure rates of sumoylation and isopeptidase cleavage in vitro, we developed an enzyme assay that is based on fluorescence resonance energy transfer (FRET). FRET is a process by which the excited state energy of a fluorescent donor molecule is transferred to an acceptor molecule. Efficient energy transfer requires very close proximity, and can therefore be used as a read-out for covalent and non-covalent protein interactions. The assay described here uses bacterially expressed and purified YFP-SUMO-1 and CFP-RanGAP1 as model substrates that are covalently coupled in the presence of recombinant SUMO E1 and E2 enzymes and ATP. Reactions of 25 microl volume, set up in 384-wells plates, give sufficient signal for analysis. Consequently, this assay requires very low amounts of recombinant proteins and allows measurement of time courses in high-throughput format.

Fluorescence Dynamics of a FRET Probe Designed for Crowding Studies.

PubMed

Currie, Megan; Leopold, Hannah; Schwarz, Jacob; Boersma, Arnold J; Sheets, Erin D; Heikal, Ahmed A

2017-06-15

Living cells are crowded with macromolecules and organelles. As a result, there is an urgent need for molecular sensors for quantitative, site-specific assessment of the macromolecular crowding effects on a myriad of biochemical processes toward quantitative cell biology and biophysics. Here we investigate the excited-state dynamics and translational diffusion of a novel FRET sensor (mCerulean-linker-mCitrine) in a buffer (PBS, pH 7.4) at room temperature. Complementary experiments were carried out on free CFP, YFP, and the cleaved FRET probe as controls. The wavelength-dependent fluorescence lifetime measurements of the donor and acceptor in the FRET probe, using the time-correlated single-photon counting technique, indicate an energy transfer efficiency of 6.8 ± 0.9% in PBS, with distinct excited-state dynamics from the recombinant CFP and YFP. The estimated mCerulean-mCitrine distance in this FRET probe is 7.7 ± 0.2 nm. The energy transfer efficiency increases (11.5 ± 0.9%) as the concentration of Ficoll-70 increases over the range of 0-300 g/L with an estimated mCerulean-mCitrine distance of 6.1 ± 0.2 nm. Complementary time-resolved anisotropy measurements suggest that the rotational diffusion of hetero-FRET in PBS is sensitive to the energy transfer from the donor to the acceptor. The results also suggest that the linker, -(GSG) 6 A(EAAAK) 6 A(GSG) 6 A(EAAAK) 6 A(GSG) 6 -, is rather flexible, and the observed rotational dynamics is likely to be due to a segmental mobility of the FRET pairs rather than an overall tumbling motion of a rigid probe. Comparative studies on a new construct of a FRET probe with a shorter, more flexible linker, mCerulean-(GSG) 18 -mCitrine, reveal enhanced energy transfer efficiency. On the millisecond time scale, fluorescence fluctuation analyses of the acceptor (excited at 488 nm) provide a means to examine the translational diffusion coefficient of the FRET probe. The results also suggest that the linker is flexible in this FRET probe, and the observed diffusion coefficient is faster than predicted as compared to the cleaved FRET probe. Our results serve as a point of reference for this FRET probe in a buffer toward its full potential as a sensor for macromolecular crowding in living cells and tissues.

Modern and Unconventional Approaches to Karst Hydrogeology

NASA Astrophysics Data System (ADS)

Sukop, M. C.

2017-12-01

Karst hydrogeology is frequently approached from a hydrograph/statistical perspective where precipitation/recharge inputs are converted to output hydrographs and the conversion process reflects the hydrology of the system. Karst catchments show hydrological response to short-term meteorological events and to long-term variation of large-scale atmospheric circulation. Modern approaches to analysis of these data include, for example, multiresolution wavelet techniques applied to understand relations between karst discharge and climate fields. Much less effort has been directed towards direct simulation of flow fields and transport phenomena in karst settings. This is primarily due to the lack of information on the detailed physical geometry of most karst systems. New mapping, sampling, and modeling techniques are beginning to enable direct simulation of flow and transport. A Conduit Flow Process (CFP) add-on to the USGS ModFlow model became available in 2007. FEFLOW and similar models are able to represent flows in individual conduits. Lattice Boltzmann models have also been applied to flow modeling in karst systems. Regarding quantitative measurement of karst system geometry, at scales to 0.1 m, X-ray computed tomography enables good detection of detailed (sub-millimeter) pore space in karstic rocks. Three-dimensional printing allows reconstruction of fragile high porosity rocks, and surrogate samples generated this way can then be subjected to laboratory testing. Borehole scales can be accessed with high-resolution ( 0.001 m) Digital Optical Borehole Imaging technologies and can provide virtual samples more representative of the true nature of karst aquifers than can obtained from coring. Subsequent extrapolation of such samples can generate three-dimensional models suitable for direct modeling of flow and transport. Finally, new cave mapping techniques are beginning to provide information than can be applied to direct simulation of flow. Due to flow rates and cave diameter, very high Reynolds number flows may be encountered.

GWM-VI: groundwater management with parallel processing for multiple MODFLOW versions

USGS Publications Warehouse

Banta, Edward R.; Ahlfeld, David P.

2013-01-01

Groundwater Management–Version Independent (GWM–VI) is a new version of the Groundwater Management Process of MODFLOW. The Groundwater Management Process couples groundwater-flow simulation with a capability to optimize stresses on the simulated aquifer based on an objective function and constraints imposed on stresses and aquifer state. GWM–VI extends prior versions of Groundwater Management in two significant ways—(1) it can be used with any version of MODFLOW that meets certain requirements on input and output, and (2) it is structured to allow parallel processing of the repeated runs of the MODFLOW model that are required to solve the optimization problem. GWM–VI uses the same input structure for files that describe the management problem as that used by prior versions of Groundwater Management. GWM–VI requires only minor changes to the input files used by the MODFLOW model. GWM–VI uses the Joint Universal Parameter IdenTification and Evaluation of Reliability Application Programming Interface (JUPITER-API) to implement both version independence and parallel processing. GWM–VI communicates with the MODFLOW model by manipulating certain input files and interpreting results from the MODFLOW listing file and binary output files. Nearly all capabilities of prior versions of Groundwater Management are available in GWM–VI. GWM–VI has been tested with MODFLOW-2005, MODFLOW-NWT (a Newton formulation for MODFLOW-2005), MF2005-FMP2 (the Farm Process for MODFLOW-2005), SEAWAT, and CFP (Conduit Flow Process for MODFLOW-2005). This report provides sample problems that demonstrate a range of applications of GWM–VI and the directory structure and input information required to use the parallel-processing capability.

Prokaryotic cells: structural organisation of the cytoskeleton and organelles.

PubMed

Souza, Wanderley de

2012-05-01

For many years, prokaryotic cells were distinguished from eukaryotic cells based on the simplicity of their cytoplasm, in which the presence of organelles and cytoskeletal structures had not been discovered. Based on current knowledge, this review describes the complex components of the prokaryotic cell cytoskeleton, including (i) tubulin homologues composed of FtsZ, BtuA, BtuB and several associated proteins, which play a fundamental role in cell division, (ii) actin-like homologues, such as MreB and Mb1, which are involved in controlling cell width and cell length, and (iii) intermediate filament homologues, including crescentin and CfpA, which localise on the concave side of a bacterium and along its inner curvature and associate with its membrane. Some prokaryotes exhibit specialised membrane-bound organelles in the cytoplasm, such as magnetosomes and acidocalcisomes, as well as protein complexes, such as carboxysomes. This review also examines recent data on the presence of nanotubes, which are structures that are well characterised in mammalian cells that allow direct contact and communication between cells.

Dependence of fluorescent protein brightness on protein concentration in solution and enhancement of it

PubMed Central

Morikawa, Takamitsu J.; Fujita, Hideaki; Kitamura, Akira; Horio, Takashi; Yamamoto, Johtaro; Kinjo, Masataka; Sasaki, Akira; Machiyama, Hiroaki; Yoshizawa, Keiko; Ichimura, Taro; Imada, Katsumi; Nagai, Takeharu; Watanabe, Tomonobu M.

2016-01-01

Fluorescent proteins have been widely used in biology because of their compatibility and varied applications in living specimens. Fluorescent proteins are often undesirably sensitive to intracellular conditions such as pH and ion concentration, generating considerable issues at times. However, harnessing these intrinsic sensitivities can help develop functional probes. In this study, we found that the fluorescence of yellow fluorescent protein (YFP) depends on the protein concentration in the solution and that this dependence can be enhanced by adding a glycine residue in to the YFP; we applied this finding to construct an intracellular protein-crowding sensor. A Förster resonance energy transfer (FRET) pair, involving a cyan fluorescent protein (CFP) insensitive to protein concentration and a glycine-inserted YFP, works as a genetically encoded probe to evaluate intracellular crowding. By measuring the fluorescence of the present FRET probe, we were able to detect dynamic changes in protein crowding in living cells. PMID:26956628

Imaging of protein kinase C activation by FRET during proliferation induced by low-energy laser irradiation in living cells

NASA Astrophysics Data System (ADS)

Gao, Xuejuan; Chen, Tongsheng; Xing, Da; Wang, Fang

2005-01-01

Protein kinase Cs (PKCs) play an important role in cellular proliferation, and low-energy laser irradiation (LELI) can enhance cellular proliferation. The present work contributes to the understanding of the mechanisms of action by studying effects of LELI at the dose of 0.8 J/cm2 on PKCs activities in the single lung adenocarcinoma cell (ASTC-a-1) and in real time by fluorescence resonance energy transfer (FRET) technique. C-kinase activity reporter (CKAR), consisting of a cyan fluorescent protein (CFP), the FHA2 phosphothreonine-binding domain, a PKC substrate sequence, and a yellow fluorescent protein (YFP), was utilized. The living cell imaging showed a decrease in FRET in the cytosol and nucleus after the cells were treated with LELI. These results suggest that PKCs could be activated by LELI throughout the cell, and the proliferation of ASTC-a-1 cells could be modulated by the activated PKCs.

Evidence of C-F-P and aromatic π-F-P weak interactions in imidazolium ionic liquids and its consequences

NASA Astrophysics Data System (ADS)

Panja, Sumit Kumar; Srivastava, Nitin; Srivastava, Jyoti; Prasad, Namburi Eswara; Noothalapati, Hemanth; Shigeto, Shinsuke; Saha, Satyen

2018-04-01

A simple change from alkyl group to alkene in side chain of imidazolium cation with same anion resulted in a drastic impact on physical properties (e.g., melting point) from bmimPF6 IL to cmimPF6 IL. The underlying reasons have been elucidated by structural and interaction studies with the help of DSC, SCXRD, vibrational and multi-nuclear NMR spectroscopic techniques. Experiments reveal existence of new weak interactions involving the carbon and π cloud of the imidazolium aromatic ring with fluoride of PF6 anion (i.e., C2-F-P and π-F-P) in cmimPF6 but are absent in structurally similar prototype IL, bmimPF6. Though weak, these interactions helped to form ladder type supramolecular arrangement, resulting in quite high melting point for cmimPF6 IL compared to bmimPF6 IL. These findings emphasize that an IL system can behave uniquely because of the existence of uncommon weak interactions.

Designing the Sniper: Improving Targeted Human Cytolytic Fusion Proteins for Anti-Cancer Therapy via Molecular Simulation.

PubMed

Bochicchio, Anna; Jordaan, Sandra; Losasso, Valeria; Chetty, Shivan; Perera, Rodrigo Casasnovas; Ippoliti, Emiliano; Barth, Stefan; Carloni, Paolo

2017-02-17

Targeted human cytolytic fusion proteins (hCFPs) are humanized immunotoxins for selective treatment of different diseases including cancer. They are composed of a ligand specifically binding to target cells genetically linked to a human apoptosis-inducing enzyme. hCFPs target cancer cells via an antibody or derivative (scFv) specifically binding to e.g., tumor associated antigens (TAAs). After internalization and translocation of the enzyme from endocytosed endosomes, the human enzymes introduced into the cytosol are efficiently inducing apoptosis. Under in vivo conditions such enzymes are subject to tight regulation by native inhibitors in order to prevent inappropriate induction of cell death in healthy cells. Tumor cells are known to upregulate these inhibitors as a survival mechanism resulting in escape of malignant cells from elimination by immune effector cells. Cytosolic inhibitors of Granzyme B and Angiogenin (Serpin P9 and RNH1, respectively), reduce the efficacy of hCFPs with these enzymes as effector domains, requiring detrimentally high doses in order to saturate inhibitor binding and rescue cytolytic activity. Variants of Granzyme B and Angiogenin might feature reduced affinity for their respective inhibitors, while retaining or even enhancing their catalytic activity. A powerful tool to design hCFPs mutants with improved potency is given by in silico methods. These include molecular dynamics (MD) simulations and enhanced sampling methods (ESM). MD and ESM allow predicting the enzyme-protein inhibitor binding stability and the associated conformational changes, provided that structural information is available. Such "high-resolution" detailed description enables the elucidation of interaction domains and the identification of sites where particular point mutations may modify those interactions. This review discusses recent advances in the use of MD and ESM for hCFP development from the viewpoints of scientists involved in both fields.

Designing the Sniper: Improving Targeted Human Cytolytic Fusion Proteins for Anti-Cancer Therapy via Molecular Simulation

PubMed Central

Bochicchio, Anna; Jordaan, Sandra; Losasso, Valeria; Chetty, Shivan; Casasnovas Perera, Rodrigo; Ippoliti, Emiliano; Barth, Stefan; Carloni, Paolo

2017-01-01

Targeted human cytolytic fusion proteins (hCFPs) are humanized immunotoxins for selective treatment of different diseases including cancer. They are composed of a ligand specifically binding to target cells genetically linked to a human apoptosis-inducing enzyme. hCFPs target cancer cells via an antibody or derivative (scFv) specifically binding to e.g., tumor associated antigens (TAAs). After internalization and translocation of the enzyme from endocytosed endosomes, the human enzymes introduced into the cytosol are efficiently inducing apoptosis. Under in vivo conditions such enzymes are subject to tight regulation by native inhibitors in order to prevent inappropriate induction of cell death in healthy cells. Tumor cells are known to up-regulate these inhibitors as a survival mechanism resulting in escape of malignant cells from elimination by immune effector cells. Cytosolic inhibitors of Granzyme B and Angiogenin (Serpin P9 and RNH1, respectively), reduce the efficacy of hCFPs with these enzymes as effector domains, requiring detrimentally high doses in order to saturate inhibitor binding and rescue cytolytic activity. Variants of Granzyme B and Angiogenin might feature reduced affinity for their respective inhibitors, while retaining or even enhancing their catalytic activity. A powerful tool to design hCFPs mutants with improved potency is given by in silico methods. These include molecular dynamics (MD) simulations and enhanced sampling methods (ESM). MD and ESM allow predicting the enzyme-protein inhibitor binding stability and the associated conformational changes, provided that structural information is available. Such “high-resolution” detailed description enables the elucidation of interaction domains and the identification of sites where particular point mutations may modify those interactions. This review discusses recent advances in the use of MD and ESM for hCFP development from the viewpoints of scientists involved in both fields. PMID:28536352

Optimizing the sensory characteristics and acceptance of canned cat food: use of a human taste panel.

PubMed

Pickering, G J

2009-02-01

A methodology based on descriptive analysis techniques used in the evaluation of human food has been successfully refined to allow for a human taste panel to profile the flavour and texture of a range of cat food products (CFP) and their component parts. Included in this method is the development of evaluation protocols for homogeneous products and for binary samples containing both meat chunk (MC) and gravy/gel (GG) constituents. Using these techniques, 18 flavour attributes (sweet, sour/acid, tuna, herbal, spicy, soy, salty, cereal, caramel, chicken, methionine, vegetable, offaly, meaty, burnt flavour, prawn, rancid and bitter) and four texture dimensions (hardness, chewiness, grittiness and viscosity) were generated to describe the sensations elicited by 13 commercial pet food samples. These samples differed in intensity for 16 of the 18 flavour attributes, which allows for individual CFP flavour profiles to be developed. Principal components analysis (PCA) could successfully discriminate between samples within the PCA space and also reveal some groupings amongst them. While many flavour attributes were weakly correlated, a large number (describing both taste and retro-nasal aroma qualities) were required to adequately differentiate between samples, suggesting considerable complexity in the products assessed. For both MC and GG, differences between samples for each of the texture dimensions were also found. For MC, grittiness appears to be the most discriminating textural attribute, while for GG viscosity discriminates well between samples. Meat chunks and gravy/gels differed significantly from each other in both flavour and texture. Cat food products differed in their liking ratings, although no differences were found between homogeneous, MC and GG samples, and eight flavour attributes were correlated with overall liking scores. It is now necessary to determine the usefulness and limits of sensory data gathered from human panels in describing and predicting food acceptance and preference behaviours in cats. For instance, while the sense of taste in cats appears generally similar to that of other mammals, they lack a sweet taste receptor (Li et al., 2006), which may limit the applicability of sweetness ratings obtained from humans. Modification of existing techniques used with human food research, such as external preference mapping (Naes and Risvik, 1996) may be useful. Ultimately, this may facilitate more economical and efficient methods for optimizing cat food flavour and texture and predicting the effects of composition and processing changes on cat feeding behaviour. This will require collaboration between pet food manufacturers and nutritionists, animal behaviourists and human sensory scientists. The results of this preliminary study should assist in this process.

Potential of DosR and Rpf antigens from Mycobacterium tuberculosis to discriminate between latent and active tuberculosis in a tuberculosis endemic population of Medellin Colombia.

PubMed

Arroyo, Leonar; Marín, Diana; Franken, Kees L M C; Ottenhoff, Tom H M; Barrera, Luis F

2018-01-08

Tuberculosis (TB) remains one of the most deadly infectious diseases. One-third to one-fourth of the human population is estimated to be infected with Mycobacterium tuberculosis (Mtb) without showing clinical symptoms, a condition called latent TB infection (LTBI). Diagnosis of Mtb infection is based on the immune response to a mixture of mycobacterial antigens (PPD) or to Mtb specific ESAT-6/CFP10 antigens (IGRA), highly expressed during the initial phase of infection. However, the immune response to PPD and IGRA antigens has a low power to discriminate between LTBI and PTB. The T-cell response to a group of so-called latency (DosR-regulon-encoded) and Resuscitation Promoting (Rpf) antigens of Mtb has been proved to be significantly higher in LTBI compared to active TB across many populations, suggesting their potential use as biomarkers to differentiate latent from active TB. PBMCs from a group LTBI (n = 20) and pulmonary TB patients (PTB, n = 21) from an endemic community for TB of the city of Medellín, Colombia, were in vitro stimulated for 7 days with DosR- (Rv1737c, Rv2029c, and Rv2628), Rpf- (Rv0867c and Rv2389c), the recombinant fusion protein ESAT-6-CFP10 (E6-C10)-, or PPD-antigen. The induced IFNγ levels detectable in the supernatants of the antigen-stimulated cells were then used to calculate specificity and sensitivity in discriminating LTBI from PTB, using different statistical approaches. IFNγ production in response to DosR and Rpf antigens was significantly higher in LTBI compared to PTB. ROC curve analyses of IFNγ production allowed differentiation of LTBI from PTB with areas under the curve higher than 0.70. Furthermore, Multiple Correspondence Analysis (MCA) revealed that LTBI is associated with higher levels of IFNγ in response to the different antigens compared to PTB. Analysis based on decision trees showed that the IFNγ levels produced in response to Rv2029c was the leading variable that best-classified disease status. Finally, logistic regression analysis predicted that IFNγ produced by PBMCs in response to E6-C10, Rv2029c, Rv0867c (RpfA) and Rv2389c (RpfA) antigens correlates best with the probability of being latently infected. The Mtb antigens E6-C10, Rv2029c (PfkB), Rv0867c (RpfA) and Rv2389c (RpfA), may be potential candidates to discriminate LTBI from PTB.

22 CFR 171.13 - Business information.

Code of Federal Regulations, 2013 CFR

2013-04-01

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22 CFR 171.13 - Business information.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 22 Foreign Relations 1 2014-04-01 2014-04-01 false Business information. 171.13 Section 171.13... PUBLIC Freedom of Information Act Provisions § 171.13 Business information. (a) Business information... section. (b) Definitions. For purposes of this section: (1) Business information means information...

22 CFR 171.13 - Business information.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 22 Foreign Relations 1 2012-04-01 2012-04-01 false Business information. 171.13 Section 171.13... PUBLIC Freedom of Information Act Provisions § 171.13 Business information. (a) Business information... section. (b) Definitions. For purposes of this section: (1) Business information means information...

22 CFR 171.13 - Business information.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 22 Foreign Relations 1 2011-04-01 2011-04-01 false Business information. 171.13 Section 171.13... PUBLIC Freedom of Information Act Provisions § 171.13 Business information. (a) Business information... section. (b) Definitions. For purposes of this section: (1) Business information means information...

6 CFR 5.8 - Business information.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 6 Domestic Security 1 2010-01-01 2010-01-01 false Business information. 5.8 Section 5.8 Domestic... Freedom of Information Act § 5.8 Business information. (a) In general. Business information obtained by... this section. (b) Definitions. For purposes of this section: (1) Business information means commercial...

6 CFR 5.8 - Business information.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 6 Domestic Security 1 2014-01-01 2014-01-01 false Business information. 5.8 Section 5.8 Domestic... Freedom of Information Act § 5.8 Business information. (a) In general. Business information obtained by... this section. (b) Definitions. For purposes of this section: (1) Business information means commercial...

6 CFR 5.8 - Business information.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 6 Domestic Security 1 2013-01-01 2013-01-01 false Business information. 5.8 Section 5.8 Domestic... Freedom of Information Act § 5.8 Business information. (a) In general. Business information obtained by... this section. (b) Definitions. For purposes of this section: (1) Business information means commercial...

6 CFR 5.8 - Business information.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 6 Domestic Security 1 2012-01-01 2012-01-01 false Business information. 5.8 Section 5.8 Domestic... Freedom of Information Act § 5.8 Business information. (a) In general. Business information obtained by... this section. (b) Definitions. For purposes of this section: (1) Business information means commercial...

6 CFR 5.8 - Business information.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 6 Domestic Security 1 2011-01-01 2011-01-01 false Business information. 5.8 Section 5.8 Domestic... Freedom of Information Act § 5.8 Business information. (a) In general. Business information obtained by... this section. (b) Definitions. For purposes of this section: (1) Business information means commercial...

76 FR 66325 - Agency Information Collection Activities: Proposed Collection, Comments Requested; E-FOIA

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-26

... Division Record Information Dissemination Section (RIDS) will be submitting the following information... collection: Records Management Division/ Record Information Dissemination Section, Federal Bureau of... requesters etc). Abstract: The Record/Information Dissemination Section (RIDS) effectively plans, develops...

76 FR 35231 - Notice of Submission of Proposed Information Collection to OMB Section 3 Business Self...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-16

...-certify that they meet the regulatory definition of a Section 3 Business. The information collected from... Proposed Information Collection to OMB Section 3 Business Self-Certification Application AGENCY: Office of... proposal. The information collected from the Section 3 Business Self- Certification Application will allow...

Directed Evolution to Engineer Monobody for FRET Biosensor Assembly and Imaging at Live-Cell Surface.

PubMed

Limsakul, Praopim; Peng, Qin; Wu, Yiqian; Allen, Molly E; Liang, Jing; Remacle, Albert G; Lopez, Tyler; Ge, Xin; Kay, Brian K; Zhao, Huimin; Strongin, Alex Y; Yang, Xiang-Lei; Lu, Shaoying; Wang, Yingxiao

2018-04-19

Monitoring enzymatic activities at the cell surface is challenging due to the poor efficiency of transport and membrane integration of fluorescence resonance energy transfer (FRET)-based biosensors. Therefore, we developed a hybrid biosensor with separate donor and acceptor that assemble in situ. The directed evolution and sequence-function analysis technologies were integrated to engineer a monobody variant (PEbody) that binds to R-phycoerythrin (R-PE) dye. PEbody was used for visualizing the dynamic formation/separation of intercellular junctions. We further fused PEbody with the enhanced CFP and an enzyme-specific peptide at the extracellular surface to create a hybrid FRET biosensor upon R-PE capture for monitoring membrane-type-1 matrix metalloproteinase (MT1-MMP) activities. This biosensor revealed asymmetric distribution of MT1-MMP activities, which were high and low at loose and stable cell-cell contacts, respectively. Therefore, directed evolution and rational design are promising tools to engineer molecular binders and hybrid FRET biosensors for monitoring molecular regulations at the surface of living cells. Copyright © 2018 Elsevier Ltd. All rights reserved.

Valproic Acid Influences MTNR1A Intracellular Trafficking and Signaling in a β-Arrestin 2-Dependent Manner.

PubMed

Hong, Ling-juan; Jiang, Quan; Long, Sen; Wang, Huan; Zhang, Ling-di; Tian, Yun; Wang, Cheng-kun; Cao, Jing-jing; Tao, Rong-rong; Huang, Ji-yun; Liao, Mei-hua; Lu, Ying-mei; Fukunaga, Kohji; Zhou, Nai-ming; Han, Feng

2016-03-01

Valproate exposure is associated with increased risks of autism spectrum disorder. To date, the mechanistic details of disturbance of melatonin receptor subtype 1 (MTNR1A) internalization upon valproate exposure remain elusive. By expressing epitope-tagged receptors (MTNR1A-EGFP) in HEK-293 and Neuro-2a cells, we recorded the dynamic changes of MTNR1A intracellular trafficking after melatonin treatment. Using time-lapse confocal microscopy, we showed in living cells that valproic acid interfered with the internalization kinetics of MTNR1A in the presence of melatonin. This attenuating effect was associated with a decrease in the phosphorylation of PKA (Thr197) and ERK (Thr202/Tyr204). VPA treatment did not alter the whole-cell currents of cells with or without melatonin. Furthermore, fluorescence resonance energy transfer imaging data demonstrated that valproic acid reduced the melatonin-initiated association between YFP-labeled β-arrestin 2 and CFP-labeled MTNR1A. Together, we suggest that valproic acid influences MTNR1A intracellular trafficking and signaling in a β-arrestin 2-dependent manner.

A genetic algorithm for a bi-objective mathematical model for dynamic virtual cell formation problem

NASA Astrophysics Data System (ADS)

Moradgholi, Mostafa; Paydar, Mohammad Mahdi; Mahdavi, Iraj; Jouzdani, Javid

2016-09-01

Nowadays, with the increasing pressure of the competitive business environment and demand for diverse products, manufacturers are force to seek for solutions that reduce production costs and rise product quality. Cellular manufacturing system (CMS), as a means to this end, has been a point of attraction to both researchers and practitioners. Limitations of cell formation problem (CFP), as one of important topics in CMS, have led to the introduction of virtual CMS (VCMS). This research addresses a bi-objective dynamic virtual cell formation problem (DVCFP) with the objective of finding the optimal formation of cells, considering the material handling costs, fixed machine installation costs and variable production costs of machines and workforce. Furthermore, we consider different skills on different machines in workforce assignment in a multi-period planning horizon. The bi-objective model is transformed to a single-objective fuzzy goal programming model and to show its performance; numerical examples are solved using the LINGO software. In addition, genetic algorithm (GA) is customized to tackle large-scale instances of the problems to show the performance of the solution method.

Conformational Dynamics inside Amino-Terminal Disease Hotspot of Ryanodine Receptor

PubMed Central

Zhong, Xiaowei; Liu, Ying; Zhu, Li; Meng, Xing; Wang, Ruiwu; Van Petegem, Filip; Wagenknecht, Terence; Wayne Chen, S. R.; Liu, Zheng

2013-01-01

Summary The N-terminal region of both skeletal and cardiac ryanodine receptor is a disease mutation hotspot. Recently, a crystal structure of the RyR1 fragment (residues 1-559) was solved. This N-terminal structure contains three separate domains, A, B, and C, and was docked into a central vestibule in a full-length RyR1 cryo-EM map. Here we reconstructed 3D cryo-EM structures of two GFP-tagged RyR2s with GFP inserted after residue Glu-310 and Ser-437, respectively. The structures of RyR2E310-GFP and RyR2S437-GFP displayed an extra mass on domain B and C, directly validating the predicted docking model. Next, we revealed domain movements in molecular dynamics flexible fitting models in both the closed and open state cryo-EM maps. To further probe the conformational changes, we generated FRET pairs by inserting CFP or YFP in two selected domains, FRET studies of three dual-insertion pairs and three co-expressed single-insertion pairs showed the dynamic structural changes within the N-terminal domains. PMID:24139989

Real-time single cell analysis of Bid cleavage and translocation in cisplatin-induced apoptosis

NASA Astrophysics Data System (ADS)

Liu, Lei; Xing, Da; Pei, Yihui; Chen, Wei R.

2007-02-01

Cancer cell apoptosis can be induced by cisplatin, an efficient anticancer agent. However, its mechanism is not fully understood. Bcl-2 homology domain (BH) 3-only proteins couple stress signals to mitochondrial apoptotic pathways. Calpain-mediated cleavage of the BH3-only protein Bid into a 14 kD truncated protein (tBid) has been implicated in cisplatin-induced apoptotic pathway. We utilized a recombinant fluorescence resonance energy transfer (FRET) Bid probe to determine the kinetics of Bid cleavage during cisplatin-induced apoptosis in ASTC-a-1 cells. The cells were also co-transfected with Bid-CFP and DsRed-Mit to dynamically detect tBid translocation. Cells showed a cleavage of the Bid-FRET probe occurring at about 4-5 h after treated with 20 µM cisplatin. Cleavage of the Bid-FRET probe coincided with a translocation of tBid from the cytosolic to the mitochondria, and the translocation lasted about 1.5 h. Using real-time single-cell analysis, we first observed the kinetics of Bid cleavage and translocation to mitochondria in living cells during cisplatin-induced apoptosis.

76 FR 69287 - National Instant Criminal Background Check System Section Agency Information Collection...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-11-08

... System Section Agency Information Collection Activities: Existing collection, comments requested the Voluntary Appeal File (VAF) Brochure ACTION: 60-Day Notice of Information Collection Under Review. The... Criminal Background Check System (NICS) Section has submitted the following information collection request...

78 FR 2992 - Agency Information Collection Activities; Proposed Collection; Comment Request; Distribution of...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-01-15

... Activities; Proposed Collection; Comment Request; Distribution of Offsite Consequence Analysis Information... Collection Request (ICR), Distribution of Offsite Consequence Analysis Information under Section 112(r)(7)(H... Air Act Section 112(r)(7); Distribution of Off-Site Consequence Analysis Information. CAA section 112...

7 CFR 6.9 - Information.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 1 2013-01-01 2013-01-01 false Information. 6.9 Section 6.9 Agriculture Office of the Secretary of Agriculture IMPORT QUOTAS AND FEES General Provisions § 6.9 Information. Persons desiring information from the Department of Agriculture regarding section 22 or section 8(a), or any action with...

7 CFR 6.9 - Information.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 1 2012-01-01 2012-01-01 false Information. 6.9 Section 6.9 Agriculture Office of the Secretary of Agriculture IMPORT QUOTAS AND FEES General Provisions § 6.9 Information. Persons desiring information from the Department of Agriculture regarding section 22 or section 8(a), or any action with...

7 CFR 6.9 - Information.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 1 2011-01-01 2011-01-01 false Information. 6.9 Section 6.9 Agriculture Office of the Secretary of Agriculture IMPORT QUOTAS AND FEES General Provisions § 6.9 Information. Persons desiring information from the Department of Agriculture regarding section 22 or section 8(a), or any action with...

7 CFR 6.9 - Information.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 1 2014-01-01 2014-01-01 false Information. 6.9 Section 6.9 Agriculture Office of the Secretary of Agriculture IMPORT QUOTAS AND FEES General Provisions § 6.9 Information. Persons desiring information from the Department of Agriculture regarding section 22 or section 8(a), or any action with...

78 FR 39003 - 60-Day Notice of Proposed Information Collection: Section 202 Supportive Housing for the Elderly...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-28

... Information Collection: Section 202 Supportive Housing for the Elderly Application Submission Requirements... Information Collection Title of Information Collection: Section 202 Supportive Housing for the Elderly... applicant eligibility and ability to develop housing for the elderly within statutory and program criteria...

7 CFR 6.9 - Information.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 1 2010-01-01 2010-01-01 false Information. 6.9 Section 6.9 Agriculture Office of the Secretary of Agriculture IMPORT QUOTAS AND FEES General Provisions § 6.9 Information. Persons desiring information from the Department of Agriculture regarding section 22 or section 8(a), or any action with...

NASA Scientific and Technical Information Standards

NASA Technical Reports Server (NTRS)

2006-01-01

This document presents general recommended standards for documenting scientific and technical information (STI) from a number of scientific and engineering disciplines. It is a companion publication to NASA SP-7084, "Grammar, Punctuation, and Capitalization: A Handbook for Technical Writers and Editors," and is intended primarily for STI personnel and publishing personnel within NASA and who support NASA STI publishing. Section 1 gives an overview of NASA STI publications. Section 2 discusses figure preparation considerations. Section 3 covers table design, and Section 4 gives information about symbols and math related to STI publishing. Section 5 covers units of measure. Section 6 discusses References, and Section 7 discusses electronic documents. Section 8 covers information related to the review of STI prior to publication; this covers both technical and dissemination review and approval, including data quality. Section 9 discusses printing and dissemination related to STI, and Section 10 gives abbreviations and acronyms used in the document.

77 FR 65395 - Notice of Submission of Proposed Information Collection to OMB: Section 8 Contract Renewal Policy...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-26

... Project- Based Section 8 Contracts AGENCY: Office of the Chief Information Officer, HUD. ACTION: Notice... through the use of appropriate automated collection techniques or other forms of information technology, e... Section 8 project-based assistance contracts are renewed. The Section 8 contract renewal process is an...

16 CFR 1101.11 - General application of provisions of section 6(b)(1).

Code of Federal Regulations, 2012 CFR

2012-01-01

... PRODUCT SAFETY ACT REGULATIONS INFORMATION DISCLOSURE UNDER SECTION 6(b) OF THE CONSUMER PRODUCT SAFETY ACT Information Subject to Notice and Analysis Provisions of Section 6(b)(1) § 1101.11 General... notice and analysis provisions of section 6(b)(1), information must meet all the following criteria: (1...

16 CFR 1101.11 - General application of provisions of section 6(b)(1).

Code of Federal Regulations, 2011 CFR

2011-01-01

... PRODUCT SAFETY ACT REGULATIONS INFORMATION DISCLOSURE UNDER SECTION 6(b) OF THE CONSUMER PRODUCT SAFETY ACT Information Subject to Notice and Analysis Provisions of Section 6(b)(1) § 1101.11 General... notice and analysis provisions of section 6(b)(1), information must meet all the following criteria: (1...

16 CFR 1101.11 - General application of provisions of section 6(b)(1).

Code of Federal Regulations, 2010 CFR

2010-01-01

... PRODUCT SAFETY ACT REGULATIONS INFORMATION DISCLOSURE UNDER SECTION 6(b) OF THE CONSUMER PRODUCT SAFETY ACT Information Subject to Notice and Analysis Provisions of Section 6(b)(1) § 1101.11 General... notice and analysis provisions of section 6(b)(1), information must meet all the following criteria: (1...

16 CFR 1101.11 - General application of provisions of section 6(b)(1).

Code of Federal Regulations, 2014 CFR

2014-01-01

... PRODUCT SAFETY ACT REGULATIONS INFORMATION DISCLOSURE UNDER SECTION 6(b) OF THE CONSUMER PRODUCT SAFETY ACT Information Subject to Notice and Analysis Provisions of Section 6(b)(1) § 1101.11 General... notice and analysis provisions of section 6(b)(1), information must meet all the following criteria: (1...

16 CFR § 1101.11 - General application of provisions of section 6(b)(1).

Code of Federal Regulations, 2013 CFR

2013-01-01

... CONSUMER PRODUCT SAFETY ACT REGULATIONS INFORMATION DISCLOSURE UNDER SECTION 6(b) OF THE CONSUMER PRODUCT SAFETY ACT Information Subject to Notice and Analysis Provisions of Section 6(b)(1) § 1101.11 General... notice and analysis provisions of section 6(b)(1), information must meet all the following criteria: (1...

5 CFR 2411.9 - Business information.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 5 Administrative Personnel 3 2013-01-01 2013-01-01 false Business information. 2411.9 Section 2411... INFORMATION § 2411.9 Business information. (a) In general. Business information obtained by the Authority from... of this section: (1) Business information means commercial or financial information obtained by the...

5 CFR 2411.9 - Business information.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Business information. 2411.9 Section 2411... INFORMATION § 2411.9 Business information. (a) In general. Business information obtained by the Authority from... of this section: (1) Business information means commercial or financial information obtained by the...

5 CFR 2411.9 - Business information.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 5 Administrative Personnel 3 2011-01-01 2011-01-01 false Business information. 2411.9 Section 2411... INFORMATION § 2411.9 Business information. (a) In general. Business information obtained by the Authority from... of this section: (1) Business information means commercial or financial information obtained by the...

5 CFR 2411.9 - Business information.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 5 Administrative Personnel 3 2012-01-01 2012-01-01 false Business information. 2411.9 Section 2411... INFORMATION § 2411.9 Business information. (a) In general. Business information obtained by the Authority from... of this section: (1) Business information means commercial or financial information obtained by the...

5 CFR 2411.9 - Business information.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 5 Administrative Personnel 3 2014-01-01 2014-01-01 false Business information. 2411.9 Section 2411... INFORMATION § 2411.9 Business information. (a) In general. Business information obtained by the Authority from... of this section: (1) Business information means commercial or financial information obtained by the...

76 FR 4408 - Service Contract Inventory and Corresponding Point of Contact Information Per Section 703 of...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-25

... Corresponding Point of Contact Information Per Section 703 of Division C of the Fiscal Year (FY) 2010... contact information per Section 743 of Division C of the FY 2010 Consolidated Appropriations Act, Public Law 111-117. FOR FURTHER INFORMATION CONTACT: Dennis Wilhite, Director, Office of Budget Execution and...

15 CFR 4.9 - Business Information.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 15 Commerce and Foreign Trade 1 2011-01-01 2011-01-01 false Business Information. 4.9 Section 4.9... of Information Act § 4.9 Business Information. (a) In general. Business information obtained by the... the purposes of this section: (1) Business information means commercial or financial information...

15 CFR 4.9 - Business Information.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 15 Commerce and Foreign Trade 1 2012-01-01 2012-01-01 false Business Information. 4.9 Section 4.9... of Information Act § 4.9 Business Information. (a) In general. Business information obtained by the... the purposes of this section: (1) Business information means commercial or financial information...

15 CFR 4.9 - Business Information.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 15 Commerce and Foreign Trade 1 2013-01-01 2013-01-01 false Business Information. 4.9 Section 4.9... of Information Act § 4.9 Business Information. (a) In general. Business information obtained by the... the purposes of this section: (1) Business information means commercial or financial information...

15 CFR 4.9 - Business Information.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 15 Commerce and Foreign Trade 1 2014-01-01 2014-01-01 false Business Information. 4.9 Section 4.9... of Information Act § 4.9 Business Information. (a) In general. Business information obtained by the... the purposes of this section: (1) Business information means commercial or financial information...

30 CFR 254.22 - What information must I include in the “Introduction and plan contents” section?

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 2 2011-07-01 2011-07-01 false What information must I include in the âIntroduction and plan contentsâ section? 254.22 Section 254.22 Mineral Resources BUREAU OF OCEAN ENERGY... Facilities § 254.22 What information must I include in the “Introduction and plan contents” section? The...

A method for calculating a land-use change carbon footprint (LUC-CFP) for agricultural commodities - applications to Brazilian beef and soy, Indonesian palm oil.

PubMed

Persson, U Martin; Henders, Sabine; Cederberg, Christel

2014-11-01

The world's agricultural system has come under increasing scrutiny recently as an important driver of global climate change, creating a demand for indicators that estimate the climatic impacts of agricultural commodities. Such carbon footprints, however, have in most cases excluded emissions from land-use change and the proposed methodologies for including this significant emissions source suffer from different shortcomings. Here, we propose a new methodology for calculating land-use change carbon footprints for agricultural commodities and illustrate this methodology by applying it to three of the most prominent agricultural commodities driving tropical deforestation: Brazilian beef and soybeans, and Indonesian palm oil. We estimate land-use change carbon footprints in 2010 to be 66 tCO2 /t meat (carcass weight) for Brazilian beef, 0.89 tCO2 /t for Brazilian soybeans, and 7.5 tCO2 /t for Indonesian palm oil, using a 10 year amortization period. The main advantage of the proposed methodology is its flexibility: it can be applied in a tiered approach, using detailed data where it is available while still allowing for estimation of footprints for a broad set of countries and agricultural commodities; it can be applied at different scales, estimating both national and subnational footprints; it can be adopted to account both for direct (proximate) and indirect drivers of land-use change. It is argued that with an increasing commercialization and globalization of the drivers of land-use change, the proposed carbon footprint methodology could help leverage the power needed to alter environmentally destructive land-use practices within the global agricultural system by providing a tool for assessing the environmental impacts of production, thereby informing consumers about the impacts of consumption and incentivizing producers to become more environmentally responsible. © 2014 John Wiley & Sons Ltd.

46 CFR 178.210 - Stability information.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 7 2014-10-01 2014-10-01 false Stability information. 178.210 Section 178.210 Shipping... information. (a) Stability information (stability details indicated on the Certificate of Inspection, a... this section. Enough stability information, including stability calculations and assumptions made to...

46 CFR 178.210 - Stability information.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 7 2013-10-01 2013-10-01 false Stability information. 178.210 Section 178.210 Shipping... information. (a) Stability information (stability details indicated on the Certificate of Inspection, a... this section. Enough stability information, including stability calculations and assumptions made to...

Information Networks and Education: An Analytic Bibliography.

ERIC Educational Resources Information Center

Pritchard, Roger

This literature review presents a broad and overall perspective on the various kinds of information networks that will be useful to educators in developing nations. There are five sections to the essay. The first section cites and briefly describes the literature dealing with library, information, and computer networks. Sections two and three…

75 FR 30051 - Notice of Proposed Information Collection: Comment Request; Section 202 Supportive Housing for...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-28

... Information Collection: Comment Request; Section 202 Supportive Housing for the Elderly Application Submission... also lists the following information: Title of Proposal: Section 202 Supportive Housing for the Elderly... assist HUD in determining applicant eligibility and ability to develop housing for the elderly within...

Information Systems: An Introduction for Adult Educators.

ERIC Educational Resources Information Center

Caldwell, Phyllis A.

In this paper, the author's primary focus is on a marketing information system and its potential importance for adult educators. The content is in seven sections. The first two sections briefly introduce information systems in general and their relevance for adult educators. The third section briefly describes general management information…

19 CFR 210.5 - Confidential business information.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 19 Customs Duties 3 2011-04-01 2011-04-01 false Confidential business information. 210.5 Section... business information. (a) Definition and submission. Confidential business information shall be defined and... law judge, or another section of this part states otherwise, confidential business information shall...

19 CFR 210.5 - Confidential business information.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 19 Customs Duties 3 2013-04-01 2013-04-01 false Confidential business information. 210.5 Section... business information. (a) Definition and submission. Confidential business information shall be defined and... law judge, or another section of this part states otherwise, confidential business information shall...

Health Information in Bosnian (bosanski)

MedlinePlus

... new window. A Expand Section After Surgery Home Care After Total Joint Replacement - bosanski (Bosnian) Bilingual PDF Health Information ... Information Translations J Expand Section Joint Disorders Home Care After Total Joint Replacement - bosanski (Bosnian) Bilingual PDF Health Information ...

76 FR 3649 - Notice of Proposed Information Collection; Comment Request; Section 3 Business Self-Certification...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-20

... developed to allow eligible businesses to self-certify that they meet the regulatory definition of a Section... definition of a Section 3 Business. The information collected from the Section 3 Business Self-Certification... Collection; Comment Request; Section 3 Business Self-Certification Application AGENCY: Office of the...

The Balanced Regulation of Hsc70 by DNJ-13 and UNC-23 Is Required for Muscle Functionality*

PubMed Central

Papsdorf, Katharina; Sacherl, Julia; Richter, Klaus

2014-01-01

The molecular chaperone Hsc70 assists in the folding of non-native proteins together with its J domain- and BAG domain-containing cofactors. In Caenorhabditis elegans, two BAG domain-containing proteins can be identified, one of them being UNC-23, whose mutation induces severe motility dysfunctions. Using reporter strains, we find that the full-length UNC-23, in contrast to C-terminal fragments, localizes specifically to the muscular attachment sites. C-terminal fragments of UNC-23 instead perform all Hsc70-related functions, like ATPase stimulation and regulation of folding activity, albeit with lower affinity than BAG-1. Interestingly, overexpression of CFP-Hsc70 can induce muscular defects in wild-type nematodes that phenocopy the knockout of its cofactor UNC-23. Strikingly, the motility dysfunction in the unc-23 mutated strain can be cured specifically by down-regulation of the antagonistic Hsc70 cochaperone DNJ-13, implying that the severe phenotype is caused by misregulation of the Hsc70 cycle. These findings point out that the balanced action of cofactors in the ATP-driven cycle of Hsc70 is crucial for the contribution of Hsc70 to muscle functionality. PMID:25053410

Comparison of Fast Neutron Detector Technologies

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

Stange, Sy; Mckigney, Edward Allen

2015-02-09

This report documents the work performed for the Department of Homeland Security Domestic Nuclear Detection O ce as the project Fast Neutron Detection Evaluation under contract HSHQDC-14-X-00022. This study was performed as a follow-on to the project Study of Fast Neutron Signatures and Measurement Techniques for SNM Detection - DNDO CFP11-100 STA-01. That work compared various detector technologies in a portal monitor con guration, focusing on a comparison between a number of fast neutron detection techniques and two standard thermal neutron detection technologies. The conclusions of the earlier work are contained in the report Comparison of Fast Neutron Detector Technologies.more » This work is designed to address questions raised about assumptions underlying the models built for the earlier project. To that end, liquid scintillators of two di erent sizes{ one a commercial, o -the-shelf (COTS) model of standard dimensions and the other a large, planer module{were characterized at Los Alamos National Laboratory. The results of those measurements were combined with the results of the earlier models to gain a more complete picture of the performance of liquid scintillator as a portal monitor technology.« less

SAW1 is required for SDSA double-strand break repair in S. cerevisiae.

PubMed

Diamante, Graciel; Phan, Claire; Celis, Angie S; Krueger, Jonas; Kelson, Eric P; Fischhaber, Paula L

2014-03-14

SAW1, coding for Saw1, is required for single-strand annealing (SSA) DNA double-strand break (DSB) repair in Saccharomycescerevisiae. Saw1 physically associates with Rad1 and Rad52 and recruits the Rad1-Rad10 endonuclease. Herein we show by fluorescence microscopy that SAW1 is similarly required for recruitment of Rad10 to sites of Synthesis-Dependent Strand Annealing (SDSA) and associates with sites of SDSA repair in a manner temporally overlapped with Rad10. The magnitude of induction of colocalized Saw1-CFP/Rad10-YFP/DSB-RFP foci in SDSA is more dramatic in S and G2 phase cells than in M phase, consistent with the known mechanism of SDSA. We observed a substantial fraction of foci in which Rad10 was localized to the repair site without Saw1, but few DSB sites that contained Saw1 without Rad10. Together these data are consistent with a model in which Saw1 recruits Rad1-Rad10 to SDSA sites, possibly even binding as a protein-protein complex, but departs the repair site in advance of Rad1-Rad10. Copyright © 2014 Elsevier Inc. All rights reserved.

The inulin-type oligosaccharides extract from morinda officinalis, a traditional Chinese herb, ameliorated behavioral deficits in an animal model of post-traumatic stress disorder.

PubMed

Qiu, Zhi-Kun; Liu, Chun-Hui; Gao, Zhuo-Wei; He, Jia-Li; Liu, Xu; Wei, Qing-Lan; Chen, Ji-Sheng

2016-10-01

Post-traumatic stress disorder (PTSD) is a severe psychiatric condition. The allopregnanolone biosynthesis has been implicated as one of the possible contributors to PTSD. Inulin-type oligosaccharides of morinda officinalis (IOMO) had been shown to be effective in the therapy of depression. However, few studies concern the anti-PTSD-like effects of IOMO. To evaluate this, the single prolonged stress (SPS) model was used in the present study. It had been shown that the behavioral deficits of SPS-treated rats were reversed by IOMO (25.0 and 50.0 mg/kg, i.p.), which reversed the increased freezing time in contextual fear paradigm (CFP) and the decreased time and entries in open arms in the elevated plus maze (EPM) test without affecting the locomotor activity in the open field (OF) test. In addition, the decreased allopregnanolone in the prefrontal cortex, hippocampus, and amygdala was reversed by IOMO (25.0 and 50.0 mg/kg, i.p.), respectively. In summary, the present study indicated that the IOMO exert anti-PTSD-like behaviors, which maybe associated with the brain allopregnanolone biosynthesis.

Granzyme B as a diagnostic marker of tuberculosis in patients with and without HIV coinfection.

PubMed

Sarkar, Pronoti; Mitra, Soumik; Pant, Priyannk; Kotwal, Aarti; Kakati, Barnali; Masih, Victor; Sindhwani, Girish; Biswas, Debasis

2016-05-01

Immunodiagnostic tests for tuberculosis (TB) are based on the estimation of interferon γ (IFN-γ) or IFN-γ-secreting CD4(+) T cells following ex vivo stimulation with ESAT6 and CFP-10. Sensitivity of these tests is likely to be compromised in CD4(+) T-cell-depleted situations, like HIV-TB coinfection. CD4(+) and CD8(+) T cells, isolated from 3 groups, viz., HIV-negative patients with active TB, HIV-TB coinfected patients, and healthy household contacts (HHCs) were cocultivated with autologous dendritic cells, and the cytokine response to rESAT6 stimulation was compared between groups in supernatants. While CD4(+) T-cell stimulation yielded significantly elevated levels of IFN-γ and interleukin 4 in HIV-negative TB patients, compared to HHCs, the levels of both these cytokines were nondiscriminatory between HIV-positive TB patients and HHCs. However, CD8(+) T-cell stimulation yielded significantly elevated granzyme B titers in both groups of patients, irrespective of HIV coinfection status. Hence, contrary to IFN-γ, granzyme B might be a useful diagnostic marker for Mycobacterium tuberculosis infection particularly in HIV coinfected patients. Copyright © 2016 Elsevier Inc. All rights reserved.

49 CFR 701.9 - Business information.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 7 2010-10-01 2010-10-01 false Business information. 701.9 Section 701.9... (AMTRAK) AMTRAK FREEDOM OF INFORMATION ACT PROGRAM § 701.9 Business information. (a) General. Business... purposes of this section, the following definitions apply: (1) Business information means commercial or...

5 CFR 1820.8 - Business information.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Business information. 1820.8 Section 1820... OF RECORDS OR TESTIMONY § 1820.8 Business information. (a) In general. Business information obtained.... For purposes of this section: (1) “Business information” means commercial or financial information...

5 CFR 2604.402 - Business information.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 5 Administrative Personnel 3 2010-01-01 2010-01-01 false Business information. 2604.402 Section... Exemptions Under FOIA § 2604.402 Business information. (a) In general. Business information provided to the... request except in accordance with this section. (b) Designation of business information. Submitters of...

34 CFR 5.11 - Business information.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 34 Education 1 2014-07-01 2014-07-01 false Business information. 5.11 Section 5.11 Education... Available to the Public § 5.11 Business information. (a) General. The Department discloses business... purposes of this section: (1) Business information means commercial or financial information obtained by...

5 CFR 2604.402 - Business information.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 5 Administrative Personnel 3 2011-01-01 2011-01-01 false Business information. 2604.402 Section... Exemptions Under FOIA § 2604.402 Business information. (a) In general. Business information provided to the... request except in accordance with this section. (b) Designation of business information. Submitters of...

49 CFR 701.9 - Business information.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 7 2013-10-01 2013-10-01 false Business information. 701.9 Section 701.9... (AMTRAK) AMTRAK FREEDOM OF INFORMATION ACT PROGRAM § 701.9 Business information. (a) General. Business... purposes of this section, the following definitions apply: (1) Business information means commercial or...

49 CFR 701.9 - Business information.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 7 2011-10-01 2011-10-01 false Business information. 701.9 Section 701.9... (AMTRAK) AMTRAK FREEDOM OF INFORMATION ACT PROGRAM § 701.9 Business information. (a) General. Business... purposes of this section, the following definitions apply: (1) Business information means commercial or...

49 CFR 701.9 - Business information.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 7 2014-10-01 2014-10-01 false Business information. 701.9 Section 701.9... (AMTRAK) AMTRAK FREEDOM OF INFORMATION ACT PROGRAM § 701.9 Business information. (a) General. Business... purposes of this section, the following definitions apply: (1) Business information means commercial or...

5 CFR 2604.402 - Business information.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 5 Administrative Personnel 3 2012-01-01 2012-01-01 false Business information. 2604.402 Section... Exemptions Under FOIA § 2604.402 Business information. (a) In general. Business information provided to the... request except in accordance with this section. (b) Designation of business information. Submitters of...

5 CFR 1820.8 - Business information.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 5 Administrative Personnel 3 2012-01-01 2012-01-01 false Business information. 1820.8 Section 1820... OF RECORDS OR TESTIMONY § 1820.8 Business information. (a) In general. Business information obtained.... For purposes of this section: (1) “Business information” means commercial or financial information...

5 CFR 2604.402 - Business information.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 5 Administrative Personnel 3 2014-01-01 2014-01-01 false Business information. 2604.402 Section... Exemptions Under FOIA § 2604.402 Business information. (a) In general. Business information provided to the... request except in accordance with this section. (b) Designation of business information. Submitters of...

34 CFR 5.11 - Business information.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 34 Education 1 2013-07-01 2013-07-01 false Business information. 5.11 Section 5.11 Education... Available to the Public § 5.11 Business information. (a) General. The Department discloses business... purposes of this section: (1) Business information means commercial or financial information obtained by...

5 CFR 1820.8 - Business information.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 5 Administrative Personnel 3 2014-01-01 2014-01-01 false Business information. 1820.8 Section 1820... OF RECORDS OR TESTIMONY § 1820.8 Business information. (a) In general. Business information obtained.... For purposes of this section: (1) “Business information” means commercial or financial information...

5 CFR 1820.8 - Business information.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 5 Administrative Personnel 3 2011-01-01 2011-01-01 false Business information. 1820.8 Section 1820... OF RECORDS OR TESTIMONY § 1820.8 Business information. (a) In general. Business information obtained.... For purposes of this section: (1) “Business information” means commercial or financial information...

34 CFR 5.11 - Business information.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 34 Education 1 2012-07-01 2012-07-01 false Business information. 5.11 Section 5.11 Education... Available to the Public § 5.11 Business information. (a) General. The Department discloses business... purposes of this section: (1) Business information means commercial or financial information obtained by...

34 CFR 5.11 - Business information.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 34 Education 1 2011-07-01 2011-07-01 false Business information. 5.11 Section 5.11 Education... Available to the Public § 5.11 Business information. (a) General. The Department discloses business... purposes of this section: (1) Business information means commercial or financial information obtained by...

26 CFR 301.6104(b)-1 - Publicity of information on certain information returns.

Code of Federal Regulations, 2010 CFR

2010-04-01

... furnishing such information, shall be a matter of public record: (1) Except as otherwise provided in section... section 501(c)(21) (black lung trusts) shall not be available for public inspection under section 6104(b... total liability for black lung benefits, the contributor's coal pricing policies, or any background...

26 CFR 301.6104(b)-1 - Publicity of information on certain information returns.

Code of Federal Regulations, 2012 CFR

2012-04-01

... furnishing such information, shall be a matter of public record: (1) Except as otherwise provided in section... section 501(c)(21) (black lung trusts) shall not be available for public inspection under section 6104(b... total liability for black lung benefits, the contributor's coal pricing policies, or any background...

26 CFR 301.6104(b)-1 - Publicity of information on certain information returns.

Code of Federal Regulations, 2011 CFR

2011-04-01

... furnishing such information, shall be a matter of public record: (1) Except as otherwise provided in section... section 501(c)(21) (black lung trusts) shall not be available for public inspection under section 6104(b... total liability for black lung benefits, the contributor's coal pricing policies, or any background...

26 CFR 301.6104(b)-1 - Publicity of information on certain information returns.

Code of Federal Regulations, 2013 CFR

2013-04-01

... furnishing such information, shall be a matter of public record: (1) Except as otherwise provided in section... section 501(c)(21) (black lung trusts) shall not be available for public inspection under section 6104(b... total liability for black lung benefits, the contributor's coal pricing policies, or any background...

26 CFR 301.6104(b)-1 - Publicity of information on certain information returns.

Code of Federal Regulations, 2014 CFR

2014-04-01

... furnishing such information, shall be a matter of public record: (1) Except as otherwise provided in section... section 501(c)(21) (black lung trusts) shall not be available for public inspection under section 6104(b... total liability for black lung benefits, the contributor's coal pricing policies, or any background...

17 CFR 230.421 - Presentation of information in prospectuses.

Code of Federal Regulations, 2010 CFR

2010-04-01

... in such fashion as to obscure any of the required information or any information necessary to keep... organization, language, and design of the front and back cover pages, the summary, and the risk factors section.... (3) In designing these sections or other sections of the prospectus, you may include pictures, logos...

17 CFR 230.421 - Presentation of information in prospectuses.

Code of Federal Regulations, 2011 CFR

2011-04-01

... in such fashion as to obscure any of the required information or any information necessary to keep... organization, language, and design of the front and back cover pages, the summary, and the risk factors section.... (3) In designing these sections or other sections of the prospectus, you may include pictures, logos...

IFLA General Conference 1988. Division of Management and Technology. Section on Information Technology; Section on Statistics; Section on Library Buildings and Equipment; Section on Conservation; Round Table on Management of Library Associations; Round Table on AV Media.

ERIC Educational Resources Information Center

International Federation of Library Associations, The Hague (Netherlands).

The 15 papers in this compilation focus on library nagement and technology, including information technology, statistics, buildings and equipment, and conservation: (1) "Information Control: OSI (Open Systems Interconnection) and Networking Strategies" (Neil McClean, United Kingdom); (2) "OSI in Australia: Potential, Planning,…

43 CFR 3900.5 - Information collection.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 43 Public Lands: Interior 2 2014-10-01 2014-10-01 false Information collection. 3900.5 Section... Management-Introduction § 3900.5 Information collection. (a) OMB has approved the information collection...) of this section lists the subpart in the rule requiring the information and its title, provides the...

30 CFR 401.4 - Information collection.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 30 Mineral Resources 2 2014-07-01 2014-07-01 false Information collection. 401.4 Section 401.4... General § 401.4 Information collection. (a) The information collection requirements contained in sections... seq. and assigned clearance number 1028-0044. The information will be used to support water related...

49 CFR 701.9 - Business information.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 7 2012-10-01 2012-10-01 false Business information. 701.9 Section 701.9... (AMTRAK) AMTRAK FREEDOM OF INFORMATION ACT PROGRAM § 701.9 Business information. (a) General. Business information held by Amtrak will be disclosed under the FOIA only under this section. (b) Definitions. For...

30 CFR 735.10 - Information collection.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 3 2011-07-01 2011-07-01 false Information collection. 735.10 Section 735.10... ADMINISTRATION AND ENFORCEMENT § 735.10 Information collection. (a) The information collection and retention...; and section 735.27 which was included in the above clearance numbers. (b) The information required by...

30 CFR 401.4 - Information collection.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 30 Mineral Resources 2 2013-07-01 2013-07-01 false Information collection. 401.4 Section 401.4... General § 401.4 Information collection. (a) The information collection requirements contained in sections... seq. and assigned clearance number 1028-0044. The information will be used to support water related...

30 CFR 735.10 - Information collection.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 3 2012-07-01 2012-07-01 false Information collection. 735.10 Section 735.10... ADMINISTRATION AND ENFORCEMENT § 735.10 Information collection. (a) The information collection and retention...; and section 735.27 which was included in the above clearance numbers. (b) The information required by...

30 CFR 955.10 - Information collection.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 3 2012-07-01 2012-07-01 false Information collection. 955.10 Section 955.10... FEDERAL PROGRAM STATES AND ON INDIAN LANDS § 955.10 Information collection. The information collection... assigned clearance number 1029-0083. This information is needed to meet the requirements of sections 504...

30 CFR 955.10 - Information collection.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 3 2011-07-01 2011-07-01 false Information collection. 955.10 Section 955.10... FEDERAL PROGRAM STATES AND ON INDIAN LANDS § 955.10 Information collection. The information collection... assigned clearance number 1029-0083. This information is needed to meet the requirements of sections 504...

National Renewable Energy Laboratory Information Resources Catalogue. A collection of energy efficiency and renewable energy information resources

NASA Astrophysics Data System (ADS)

1994-05-01

NREL's first annual Information Resources Catalogue is intended to inform anyone interested in energy efficiency and renewable energy technologies of NREL's outreach activities, including publications and services. For ease of use, all entries are categorized by subject. The catalogue is separated into six main sections. The first section lists and describes services that are available through NREL and how they may be accessed. The second section contains a list of documents that are published by NREL on a regular or periodic basis. The third section highlights NREL's series publications written for specific audiences and presenting a wide range of subjects. NREL's General Interest Publications constitute the fourth section of the catalogue and are written for nontechnical audiences. Descriptions are provided for these publications. The fifth section contains Technical Reports that detail research and development projects. The section on Conference Papers/Journal Articles/Book Chapters makes up the sixth and final section of the catalogue.

PPIS Information and Report Samples

EPA Pesticide Factsheets

Technical information about the Pesticide Product Information System and its downloadable file formats, and sample reporting for Federal Insecticide Fungicide and Rodenticide Act (FIFRA) section 3 and section 24(c).

NASA Publications Guide for Authors

NASA Technical Reports Server (NTRS)

2015-01-01

This document presents guidelines for use by NASA authors in preparation and publication of their scientific and technical information (STI). Section 2 gives an overview. Section 2 describes types of publication. Section 3 discusses technical, data/information, and dissemination reviews. Section 4 provides recommended standards and gives the elements of a typical report. Section 5 presents miscellaneous preparation recommendations.

29 CFR 70.26 - Business information.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 1 2010-07-01 2010-07-01 true Business information. 70.26 Section 70.26 Labor Office of... of Records Under the Freedom of Information Act § 70.26 Business information. (a) In general. Confidential business information will be disclosed under the FOIA only in accordance with this section. (b...

30 CFR 822.10 - Information collection.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 3 2010-07-01 2010-07-01 false Information collection. 822.10 Section 822.10... ALLUVIAL VALLEY FLOORS § 822.10 Information collection. The information collection requirements contained... sections 510(b)(5) and 515(b)(10)(F) of the Act which provide the information collection requirements and...

49 CFR 232.21 - Information Collection.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 4 2010-10-01 2010-10-01 false Information Collection. 232.21 Section 232.21... EQUIPMENT; END-OF-TRAIN DEVICES General § 232.21 Information Collection. (a) The information collection... information collection requirements are found in the following sections: §§ 229.27, 231.31, 232.1, 232.3, 232...

29 CFR 70.26 - Business information.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 1 2011-07-01 2011-07-01 false Business information. 70.26 Section 70.26 Labor Office of... of Records Under the Freedom of Information Act § 70.26 Business information. (a) In general. Confidential business information will be disclosed under the FOIA only in accordance with this section. (b...

29 CFR 70.26 - Business information.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 1 2014-07-01 2013-07-01 true Business information. 70.26 Section 70.26 Labor Office of... of Records Under the Freedom of Information Act § 70.26 Business information. (a) In general. Confidential business information will be disclosed under the FOIA only in accordance with this section. (b...

29 CFR 70.26 - Business information.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 29 Labor 1 2013-07-01 2013-07-01 false Business information. 70.26 Section 70.26 Labor Office of... of Records Under the Freedom of Information Act § 70.26 Business information. (a) In general. Confidential business information will be disclosed under the FOIA only in accordance with this section. (b...

32 CFR 2402.6 - Business information.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 32 National Defense 6 2014-07-01 2014-07-01 false Business information. 2402.6 Section 2402.6... REGULATIONS IMPLEMENTING THE FREEDOM OF INFORMATION ACT § 2402.6 Business information. (a) In general. Business information obtained by OSTP from a submitter will be disclosed under FOIA only under this section...

29 CFR 70.26 - Business information.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 29 Labor 1 2012-07-01 2012-07-01 false Business information. 70.26 Section 70.26 Labor Office of... of Records Under the Freedom of Information Act § 70.26 Business information. (a) In general. Confidential business information will be disclosed under the FOIA only in accordance with this section. (b...

40 CFR 2.302 - Special rules governing certain information obtained under the Clean Water Act.

Code of Federal Regulations, 2014 CFR

2014-07-01

... provide the information was issued under section 309(a)(3) of the Act, 33 U.S.C. 1319(a)(3), whether a civil action was brought under section 309(b) of the Act, 33 U.S.C. 1319(b), and whether the information... specifically does not apply to information obtained under section 310(d) or 312(g)(3) of the Act, 33 U.S.C...

40 CFR 2.302 - Special rules governing certain information obtained under the Clean Water Act.

Code of Federal Regulations, 2012 CFR

2012-07-01

... provide the information was issued under section 309(a)(3) of the Act, 33 U.S.C. 1319(a)(3), whether a civil action was brought under section 309(b) of the Act, 33 U.S.C. 1319(b), and whether the information... specifically does not apply to information obtained under section 310(d) or 312(g)(3) of the Act, 33 U.S.C...

40 CFR 2.302 - Special rules governing certain information obtained under the Clean Water Act.

Code of Federal Regulations, 2011 CFR

2011-07-01

... provide the information was issued under section 309(a)(3) of the Act, 33 U.S.C. 1319(a)(3), whether a civil action was brought under section 309(b) of the Act, 33 U.S.C. 1319(b), and whether the information... specifically does not apply to information obtained under section 310(d) or 312(g)(3) of the Act, 33 U.S.C...

Anemia - Multiple Languages

MedlinePlus

... Section Anemia - العربية (Arabic) Bilingual PDF Health Information Translations Bosnian (bosanski) Expand Section Anemia - bosanski (Bosnian) Bilingual PDF Health Information Translations Chinese, Simplified (Mandarin dialect) (简体中文) Expand Section Anemia - ...

Determining Primary Care Physician Information Needs to Inform Ambulatory Visit Note Display

PubMed Central

Clarke, M.A.; Steege, L.M.; Moore, J.L.; Koopman, R.J.; Belden, J.L.; Kim, M.S.

2014-01-01

Summary Background With the increase in the adoption of electronic health records (EHR) across the US, primary care physicians are experiencing information overload. The purpose of this pilot study was to determine the information needs of primary care physicians (PCPs) as they review clinic visit notes to inform EHR display. Method Data collection was conducted with 15 primary care physicians during semi-structured interviews, including a third party observer to control bias. Physicians reviewed major sections of an artificial but typical acute and chronic care visit note to identify the note sections that were relevant to their information needs. Statistical methods used were McNemar-Mosteller’s and Cochran Q. Results Physicians identified History of Present Illness (HPI), Assessment, and Plan (A&P) as the most important sections of a visit note. In contrast, they largely judged the Review of Systems (ROS) to be superfluous. There was also a statistical difference in physicians’ highlighting among all seven major note sections in acute (p = 0.00) and chronic (p = 0.00) care visit notes. Conclusion A&P and HPI sections were most frequently identified as important which suggests that physicians may have to identify a few key sections out of a long, unnecessarily verbose visit note. ROS is viewed by doctors as mostly “not needed,” but can have relevant information. The ROS can contain information needed for patient care when other sections of the Visit note, such as the HPI, lack the relevant information. Future studies should include producing a display that provides only relevant information to increase physician efficiency at the point of care. Also, research on moving A&P to the top of visit notes instead of having A&P at the bottom of the page is needed, since those are usually the first sections physicians refer to and reviewing from top to bottom may cause cognitive load. PMID:24734131

78 FR 40635 - Delegation of Authority to the Southern Ute Indian Tribe To Implement and Enforce National...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-07-08

....regulations.gov Web site is an ``anonymous access'' system, which means EPA will not know your identity or... submitting comments, go to Section I. General Information of the SUPPLEMENTARY INFORMATION section of this... taken under CAA sections 111 and 112. Table of Contents I. General Information II. Delegation of...

A bibliography of all known publications & reports on the Gulf sturgeon, Acipenser oxyrinchus desotoi

USGS Publications Warehouse

Price, Melissa; Adler, Jennifer; Littles, Chanda; Randolph, April Norem; Nash, Ursula A.; Gillett, Bethan; Randall, Michael T.; Sulak, Kenneth J.; Walsh, Stephen J.; Brownell, Prescott

2013-01-01

This functional bibliography is meant to be a complete and comprehensive bibliography of all discoverable reports containing information on the Gulf Sturgeon (GS). This bibliography contains all known reports presenting, documenting, summarizing, listing, or interpreting information on the GS through 31 December 2013. Report citations are organized into four sections. Section I includes published scientific journal articles, books, dissertations and theses, published and unpublished technical reports, published harvest prohibitions, and online articles reporting substantive scientific information. Section II includes newspaper, newsletter, magazine, book, agency news releases, and online articles reporting on GS occurrences, mortalities, captures, jumping, boat collisions, aquaculture, historical photographs, and other largely non-scientific or anecdotal issues. Section III consists of books, theses, ecotour-guides, media articles, editorials, and blogs reporting a mix of anecdotal information, historical information, and opinion on GS conservation, habitat issues, exploitation, aquaculture, and human interaction - but presenting very limited or no substantive scientific information. Section IV includes videos, films and audio recordings documenting GS life history and behavior.

High Temperature Reactor (HTR) Deep Burn Core and Fuel Analysis: Design Selection for the Prismatic Block Reactor With Results from FY-2011 Activities

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

Michael A. Pope

2011-10-01

The Deep Burn (DB) Project is a U.S. Department of Energy sponsored feasibility study of Transuranic Management using high burnup fuel in the high temperature helium cooled reactor (HTR). The DB Project consists of seven tasks: project management, core and fuel analysis, spent fuel management, fuel cycle integration, TRU fuel modeling, TRU fuel qualification, and HTR fuel recycle. In the Phase II of the Project, we conducted nuclear analysis of TRU destruction/utilization in the HTR prismatic block design (Task 2.1), deep burn fuel/TRISO microanalysis (Task 2.3), and synergy with fast reactors (Task 4.2). The Task 2.1 covers the core physicsmore » design, thermo-hydraulic CFD analysis, and the thermofluid and safety analysis (low pressure conduction cooling, LPCC) of the HTR prismatic block design. The Task 2.3 covers the analysis of the structural behavior of TRISO fuel containing TRU at very high burnup level, i.e. exceeding 50% of FIMA. The Task 4.2 includes the self-cleaning HTR based on recycle of HTR-generated TRU in the same HTR. Chapter IV contains the design and analysis results of the 600MWth DB-HTR core physics with the cycle length, the average discharged burnup, heavy metal and plutonium consumptions, radial and axial power distributions, temperature reactivity coefficients. Also, it contains the analysis results of the 450MWth DB-HTR core physics and the analysis of the decay heat of a TRU loaded DB-HTR core. The evaluation of the hot spot fuel temperature of the fuel block in the DB-HTR (Deep-Burn High Temperature Reactor) core under full operating power conditions are described in Chapter V. The investigated designs are the 600MWth and 460MWth DB-HTRs. In Chapter VI, the thermo-fluid and safety of the 600MWth DB-HTRs has been analyzed to investigate a thermal-fluid design performance at the steady state and a passive safety performance during an LPCC event. Chapter VII describes the analysis results of the TRISO fuel microanalysis of the 600MWth and 450MWth DB-HTRs. The TRISO fuel microanalysis covers the gas pressure buildup in a coated fuel particle including helium production, the thermo-mechanical behavior of a CFP, the failure probabilities of CFPs, the temperature distribution in a CPF, and the fission product (FP) transport in a CFP and a graphite. In Chapter VIII, it contains the core design and analysis of sodium cooled fast reactor (SFR) with deep burn HTR reactor. It considers a synergistic combination of the DB-MHR and an SFR burner for a safe and efficient transmutation of the TRUs from LWRs. Chapter IX describes the design and analysis results of the self-cleaning (or self-recycling) HTR core. The analysis is considered zero and 5-year cooling time of the spent LWR fuels.« less

High Temperature Reactor (HTR) Deep Burn Core and Fuel Analysis: Design Selection for the Prismatic Block Reactor

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

Francesco Venneri; Chang-Keun Jo; Jae-Man Noh

2010-09-01

The Deep Burn (DB) Project is a U.S. Department of Energy sponsored feasibility study of Transuranic Management using high burnup fuel in the high temperature helium cooled reactor (HTR). The DB Project consists of seven tasks: project management, core and fuel analysis, spent fuel management, fuel cycle integration, TRU fuel modeling, TRU fuel qualification, and HTR fuel recycle. In the Phase II of the Project, we conducted nuclear analysis of TRU destruction/utilization in the HTR prismatic block design (Task 2.1), deep burn fuel/TRISO microanalysis (Task 2.3), and synergy with fast reactors (Task 4.2). The Task 2.1 covers the core physicsmore » design, thermo-hydraulic CFD analysis, and the thermofluid and safety analysis (low pressure conduction cooling, LPCC) of the HTR prismatic block design. The Task 2.3 covers the analysis of the structural behavior of TRISO fuel containing TRU at very high burnup level, i.e. exceeding 50% of FIMA. The Task 4.2 includes the self-cleaning HTR based on recycle of HTR-generated TRU in the same HTR. Chapter IV contains the design and analysis results of the 600MWth DB-HTR core physics with the cycle length, the average discharged burnup, heavy metal and plutonium consumptions, radial and axial power distributions, temperature reactivity coefficients. Also, it contains the analysis results of the 450MWth DB-HTR core physics and the analysis of the decay heat of a TRU loaded DB-HTR core. The evaluation of the hot spot fuel temperature of the fuel block in the DB-HTR (Deep-Burn High Temperature Reactor) core under full operating power conditions are described in Chapter V. The investigated designs are the 600MWth and 460MWth DB-HTRs. In Chapter VI, the thermo-fluid and safety of the 600MWth DB-HTRs has been analyzed to investigate a thermal-fluid design performance at the steady state and a passive safety performance during an LPCC event. Chapter VII describes the analysis results of the TRISO fuel microanalysis of the 600MWth and 450MWth DB-HTRs. The TRISO fuel microanalysis covers the gas pressure buildup in a coated fuel particle including helium production, the thermo-mechanical behavior of a CFP, the failure probabilities of CFPs, the temperature distribution in a CPF, and the fission product (FP) transport in a CFP and a graphite. In Chapter VIII, it contains the core design and analysis of sodium cooled fast reactor (SFR) with deep burn HTR reactor. It considers a synergistic combination of the DB-MHR and an SFR burner for a safe and efficient transmutation of the TRUs from LWRs. Chapter IX describes the design and analysis results of the self-cleaning (or self-recycling) HTR core. The analysis is considered zero and 5-year cooling time of the spent LWR fuels.« less

Using MODFLOW with CFP to understand conduit-matrix exchange in a karst aquifer during flooding

NASA Astrophysics Data System (ADS)

Spellman, P.; Screaton, E.; Martin, J. B.; Gulley, J.; Brown, A.

2011-12-01

Karst springs may reverse flow when allogenic runoff increases river stage faster than groundwater heads and may exchange of surface water with groundwater in the surrounding aquifer matrix. Recharged flood water is rich in nutrients, metals, and organic matter and is undersaturated with respect to calcite. Understanding the physical processes controlling this exchange of water is critical to understanding metal cycling, redox chemistry and dissolution in the subsurface. Ultimately the magnitude of conduit-matrix exchange should be governed by head gradients between the conduit and the aquifer which are affected by the hydraulic conductivity of the matrix, conduit properties and antecedent groundwater heads. These parameters are interrelated and it is unknown which ones exert the greatest control over the magnitude of exchange. This study uses MODFLOW-2005 coupled with the Conduit Flow Processes (CFP) package to determine how physical properties of conduits and aquifers influence the magnitude of surface water-groundwater exchange. We use hydraulic data collected during spring reversals in a mapped underwater cave that sources Madison Blue Spring in north-central Florida to explore which factors are most important in governing exchange. The simulation focused on a major flood in 2009, when river stage increased by about 10 meters over 9 days. In a series of simulations, we varied hydraulic conductivity, conduit diameter, roughness height and tortuosity in addition to antecedent groundwater heads to estimate the relative effects of each parameter on the magnitude of conduit-matrix exchange. Each parameter was varied across plausible ranges for karst aquifers. Antecedent groundwater heads were varied using well data recorded through wet and dry seasons throughout the spring shed. We found hydraulic conductivity was the most important factor governing exchange. The volume of exchange increased by about 61% from the lowest value (1.8x10-6 m/d) to the highest value (6 m/d) of matrix hydraulic conductivity. Other factors increased the amount of exchange by 1% or less, with tortuosity (which varied from 1 to 2) being most significant with a 1% increase, followed by conduit diameter (1 to 5 m) and roughness height (0.1 to 5m) with increases in exchange of 0.4% and 0.3% respectively. Antecedent aquifer conditions were also seen to exert important controls on influencing exchange with greater exchange occurring in floods following dry periods than during wet periods. These preliminary results indicate that heterogeneity of the hydraulic conductivity across karst aquifers will control the distribution of flood waters that enter into the aquifer matrix. Because flood waters are typically undersaturated with respect to the carbonate minerals, the location of this infiltrated water into the highest hydraulic conductivity zones should enhance dissolution, thereby increasing hydraulic conductivity in a feedback loop that will enhance future infiltration of floodwater. Portions of the aquifer prone to infiltrating flood water and dissolution will also be most sensitive to contamination from surface water infiltration.

Bio-Oil Separation and Stabilization by Near-Critical Propane Fractionation

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

Ginosar, Daniel M.; Petkovic, Lucia M.; Agblevor, Foster A.

Bio-oils produced by thermal process are promising sources of sustainable, low greenhouse gas alternative fuels. These thermal processes are also well suited to decentralized energy production due to low capital and operating costs. Algae feedstocks for bio-oil production are of particular interest, due in part to their high-energy growth yields. Further, algae can be grown in non-arable areas in fresh, brackish, salt water, or even waste water. Unfortunately, bio-oils produced by thermal processes present significant stability challenges. These oils have complex chemical compositions, are viscous, reactive, and thermally unstable. Further, the components within the oils are difficult to separate bymore » fractional distillation. By far, the most effective separation and stabilization method has been solvent extraction. However, liquid phase extraction processes pose two main obstacles to commercialization; they require a significant amount of energy to remove and recover the solvent from the product, and they have a propensity for the solvent to become contaminated with minerals from the char and ash present in the original bio-oil. Separation and fractionation of thermally produced bio-oils using supercritical fluids (SCF) offers the advantages of liquid solvent extraction while drastically reducing energy demands and the predisposition to carry over solids into the extracted phase. SCFs are dense fluids with liquid-like solvent properties and gas-like transport properties. Further, SCF density and solvent strength can be tuned with minor adjustments in pressure, co-solvent addition, or gas anti-solvent addition. Catalytic pyrolysis oils were produced from Scenedesmus dimorphus algae using a fluid catalytic cracking catalyst. Bio-oil produced from catalytic fast pyrolysis (CFP) was separated using critical fluids. Propane extraction was performed at 65 °C at a fluid reduced pressure of 2.0 (85 bar) using an eight to one solvent to feed ratio by weight. Extraction of catalytic fast pyrolysis oil with near critical propane produced an oil extract that was physically and chemically different from and more stable than the original oil. The propane extract displayed lower viscosity and lower average molecular weight. The species present in the propane extract were likely the less polar that would be expected from using a non-polar solvent (propane). Carbonyl containing species in the extract were likely ketones and esters. The raffinate contained a higher amnount of OH bonded species along with the more polar more polar acids, amides, and alcohols. The higher concentration of nitrogen in the raffinate may confirm the presence of amides. Viscosity of the propane extract increased only half as much as that of the CFP bio-oil. Further, In situ NMR aging studies showed that the propane extract was more stable than the raw oil. In conclusion, propane extraction is a promising method to decrease the nitrogen content of bio-oils and to improve the stability of bio-oils obtained by the catalytic pyrolysis of algae based biomass.« less

18 CFR 388.113 - Accessing critical energy infrastructure information.

Code of Federal Regulations, 2014 CFR

2014-04-01

... energy infrastructure information. 388.113 Section 388.113 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY PROCEDURAL RULES INFORMATION AND REQUESTS § 388.113 Accessing critical energy infrastructure information. (a) Scope. This section governs access to...

18 CFR 388.113 - Accessing critical energy infrastructure information.

Code of Federal Regulations, 2010 CFR

2010-04-01

... energy infrastructure information. 388.113 Section 388.113 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY PROCEDURAL RULES INFORMATION AND REQUESTS § 388.113 Accessing critical energy infrastructure information. (a) Scope. This section governs access to...

18 CFR 388.113 - Accessing critical energy infrastructure information.

Code of Federal Regulations, 2012 CFR

2012-04-01

... energy infrastructure information. 388.113 Section 388.113 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY PROCEDURAL RULES INFORMATION AND REQUESTS § 388.113 Accessing critical energy infrastructure information. (a) Scope. This section governs access to...

18 CFR 388.113 - Accessing critical energy infrastructure information.

Code of Federal Regulations, 2013 CFR

2013-04-01

... energy infrastructure information. 388.113 Section 388.113 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY PROCEDURAL RULES INFORMATION AND REQUESTS § 388.113 Accessing critical energy infrastructure information. (a) Scope. This section governs access to...

18 CFR 388.113 - Accessing critical energy infrastructure information.

Code of Federal Regulations, 2011 CFR

2011-04-01

... energy infrastructure information. 388.113 Section 388.113 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY PROCEDURAL RULES INFORMATION AND REQUESTS § 388.113 Accessing critical energy infrastructure information. (a) Scope. This section governs access to...

77 FR 26605 - Agency Information Collection Activities: Proposed Information Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-04

... about the accuracy of information contained in a consumer report based on a direct request from a... Integrity of Information Furnished to Consumer Reporting Agencies under Section 312 of the Fair and Accurate... Accuracy and Integrity of Information Furnished to Consumer Reporting Agencies under Section 312 of the...

30 CFR 254.23 - What information must I include in the “Emergency response action plan” section?

Code of Federal Regulations, 2013 CFR

2013-07-01

... LOCATED SEAWARD OF THE COAST LINE Oil-Spill Response Plans for Outer Continental Shelf Facilities § 254.23 What information must I include in the “Emergency response action plan” section? The “Emergency response action plan” section is the core of the response plan. Put information in easy-to-use formats such...

30 CFR 254.23 - What information must I include in the “Emergency response action plan” section?

Code of Federal Regulations, 2011 CFR

2011-07-01

... FOR FACILITIES LOCATED SEAWARD OF THE COAST LINE Oil-Spill Response Plans for Outer Continental Shelf Facilities § 254.23 What information must I include in the “Emergency response action plan” section? The “Emergency response action plan”section is the core of the response plan. Put information in easy-to-use...

30 CFR 254.23 - What information must I include in the “Emergency response action plan” section?

Code of Federal Regulations, 2014 CFR

2014-07-01

... LOCATED SEAWARD OF THE COAST LINE Oil-Spill Response Plans for Outer Continental Shelf Facilities § 254.23 What information must I include in the “Emergency response action plan” section? The “Emergency response action plan” section is the core of the response plan. Put information in easy-to-use formats such...

30 CFR 254.23 - What information must I include in the “Emergency response action plan” section?

Code of Federal Regulations, 2012 CFR

2012-07-01

... LOCATED SEAWARD OF THE COAST LINE Oil-Spill Response Plans for Outer Continental Shelf Facilities § 254.23 What information must I include in the “Emergency response action plan” section? The “Emergency response action plan” section is the core of the response plan. Put information in easy-to-use formats such...

75 FR 53334 - Notice of Filing of Plats of Survey; Montana

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-31

... River, downstream, through sections 7 and 18, a portion of the subdivision of sections 7 and 18, a... present left bank of the Missouri River and informative traverse, downstream, through sections 7 and 18, the limits of erosion and informative traverse, downstream through sections 7 and 18 and certain...

76 FR 4703 - Proposed Information Collection Activity; Comment Request Proposed Projects:

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-26

... Information Collection Activity; Comment Request Proposed Projects: Title: Computerized Support Enforcement Systems. OMB No. 0980-0271. Description: The information being collected is mandated by Section 454(16) of...) approved under section 452(d) of the title, of a statewide automated data processing and information...

49 CFR 1105.3 - Information and assistance.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 8 2014-10-01 2014-10-01 false Information and assistance. 1105.3 Section 1105.3... Information and assistance. Information and assistance regarding the rules and the Board's environmental and historic review process is available by writing or calling the Section of Environmental Analysis, Surface...

49 CFR 1105.3 - Information and assistance.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 8 2010-10-01 2010-10-01 false Information and assistance. 1105.3 Section 1105.3... Information and assistance. Information and assistance regarding the rules and the Board's environmental and historic review process is available by writing or calling the Section of Environmental Analysis, Surface...

49 CFR 1105.3 - Information and assistance.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 8 2013-10-01 2013-10-01 false Information and assistance. 1105.3 Section 1105.3... Information and assistance. Information and assistance regarding the rules and the Board's environmental and historic review process is available by writing or calling the Section of Environmental Analysis, Surface...

32 CFR 1803.11 - Preliminary information.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 32 National Defense 6 2013-07-01 2013-07-01 false Preliminary information. 1803.11 Section 1803.11... REQUESTS FOR MANDATORY DECLASSIFICATION REVIEW OF CLASSIFIED INFORMATION PURSUANT TO SECTION 3.6 OF... information. Members of the public shall address all communications to the point of contact specified above...

78 FR 76832 - Information Collection Being Reviewed by the Federal Communications Commission

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-19

... 18, 2014. If you anticipate that you will be submitting comments, but find it difficult to do so... Emergency Information; Section 79.105, Video Description and Emergency Information Accessibility Requirements for All Apparatus; Section 79.106, Video Description and Emergency Information Accessibility...

17 CFR 230.168 - Exemption from sections 2(a)(10) and 5(c) of the Act for certain communications of regularly...

Code of Federal Regulations, 2011 CFR

2011-04-01

... and forward-looking information. 230.168 Section 230.168 Commodity and Securities Exchanges SECURITIES... factual business information and forward-looking information. Preliminary Notes to § 230.168. 1. This... provides a non-exclusive safe harbor for factual business information and forward-looking information...

7 CFR 277.18 - Establishment of an Automated Data Processing (ADP) and Information Retrieval System.

Code of Federal Regulations, 2012 CFR

2012-01-01

...) and Information Retrieval System. 277.18 Section 277.18 Agriculture Regulations of the Department of... Data Processing (ADP) and Information Retrieval System. (a) Scope and application. This section... costs of planning, design, development or installation of ADP and information retrieval systems if the...

7 CFR 277.18 - Establishment of an Automated Data Processing (ADP) and Information Retrieval System.

Code of Federal Regulations, 2014 CFR

2014-01-01

...) and Information Retrieval System. 277.18 Section 277.18 Agriculture Regulations of the Department of... Data Processing (ADP) and Information Retrieval System. (a) Scope and application. This section... costs of planning, design, development or installation of ADP and information retrieval systems if the...

7 CFR 277.18 - Establishment of an Automated Data Processing (ADP) and Information Retrieval System.

Code of Federal Regulations, 2011 CFR

2011-01-01

...) and Information Retrieval System. 277.18 Section 277.18 Agriculture Regulations of the Department of... Data Processing (ADP) and Information Retrieval System. (a) Scope and application. This section... costs of planning, design, development or installation of ADP and information retrieval systems if the...

7 CFR 277.18 - Establishment of an Automated Data Processing (ADP) and Information Retrieval System.

Code of Federal Regulations, 2013 CFR

2013-01-01

...) and Information Retrieval System. 277.18 Section 277.18 Agriculture Regulations of the Department of... Data Processing (ADP) and Information Retrieval System. (a) Scope and application. This section... costs of planning, design, development or installation of ADP and information retrieval systems if the...

39 CFR 267.5 - National Security Information.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 39 Postal Service 1 2012-07-01 2012-07-01 false National Security Information. 267.5 Section 267.5 Postal Service UNITED STATES POSTAL SERVICE ORGANIZATION AND ADMINISTRATION PROTECTION OF INFORMATION § 267.5 National Security Information. (a) Purpose and scope. The purpose of this section is to provide...

39 CFR 267.5 - National Security Information.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 39 Postal Service 1 2010-07-01 2010-07-01 false National Security Information. 267.5 Section 267.5 Postal Service UNITED STATES POSTAL SERVICE ORGANIZATION AND ADMINISTRATION PROTECTION OF INFORMATION § 267.5 National Security Information. (a) Purpose and scope. The purpose of this section is to provide...

39 CFR 267.5 - National Security Information.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 39 Postal Service 1 2014-07-01 2014-07-01 false National Security Information. 267.5 Section 267.5 Postal Service UNITED STATES POSTAL SERVICE ORGANIZATION AND ADMINISTRATION PROTECTION OF INFORMATION § 267.5 National Security Information. (a) Purpose and scope. The purpose of this section is to provide...

39 CFR 267.5 - National Security Information.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 39 Postal Service 1 2013-07-01 2013-07-01 false National Security Information. 267.5 Section 267.5 Postal Service UNITED STATES POSTAL SERVICE ORGANIZATION AND ADMINISTRATION PROTECTION OF INFORMATION § 267.5 National Security Information. (a) Purpose and scope. The purpose of this section is to provide...

39 CFR 267.5 - National Security Information.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 39 Postal Service 1 2011-07-01 2011-07-01 false National Security Information. 267.5 Section 267.5 Postal Service UNITED STATES POSTAL SERVICE ORGANIZATION AND ADMINISTRATION PROTECTION OF INFORMATION § 267.5 National Security Information. (a) Purpose and scope. The purpose of this section is to provide...

42 CFR 435.948 - Requesting information.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 42 Public Health 4 2010-10-01 2010-10-01 false Requesting information. 435.948 Section 435.948... Verification Requirements § 435.948 Requesting information. (a) Except as provided in paragraphs (d), (e), and (f) of this section, the agency must request information from the sources specified in this paragraph...

20 CFR 902.3 - Published information.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Published information. 902.3 Section 902.3 Employees' Benefits JOINT BOARD FOR THE ENROLLMENT OF ACTUARIES RULES REGARDING AVAILABILITY OF INFORMATION § 902.3 Published information. (a) Federal Register. Pursuant to sections 552 and 553 of title 5 of the...

19 CFR 207.30 - Comment on information.

Code of Federal Regulations, 2014 CFR

2014-04-01

... information. (a) In any final phase of an investigation under section 705 or section 735 of the Act, the... day. Comments shall only concern such information, and shall not exceed 15 pages of textual material... Act, and with respect to changes in bracketing of business proprietary information in the comments...

19 CFR 207.30 - Comment on information.

Code of Federal Regulations, 2013 CFR

2013-04-01

... information. (a) In any final phase of an investigation under section 705 or section 735 of the Act, the... day. Comments shall only concern such information, and shall not exceed 15 pages of textual material... Act, and with respect to changes in bracketing of business proprietary information in the comments...

19 CFR 207.30 - Comment on information.

Code of Federal Regulations, 2012 CFR

2012-04-01

... information. (a) In any final phase of an investigation under section 705 or section 735 of the Act, the... day. Comments shall only concern such information, and shall not exceed 15 pages of textual material... Act, and with respect to changes in bracketing of business proprietary information in the comments...

15 CFR 30.6 - Electronic Export Information data elements.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 15 Commerce and Foreign Trade 1 2012-01-01 2012-01-01 false Electronic Export Information data elements. 30.6 Section 30.6 Commerce and Foreign Trade Regulations Relating to Commerce and Foreign Trade... Electronic Export Information data elements. The information specified in this section is required for...

15 CFR 30.6 - Electronic Export Information data elements.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 15 Commerce and Foreign Trade 1 2013-01-01 2013-01-01 false Electronic Export Information data elements. 30.6 Section 30.6 Commerce and Foreign Trade Regulations Relating to Commerce and Foreign Trade... Electronic Export Information data elements. The information specified in this section is required for...

76 FR 5296 - Safety Management System for Part 121 Certificate Holders; Extension of Comment Period

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-31

... INFORMATION: See the ``Additional Information'' section for information on how to comment on this proposal and how the FAA will handle comments received. The ``Additional Information'' section also contains... comments relating to the economic, environmental, energy, or federalism impacts that might result from...

20 CFR 902.3 - Published information.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 20 Employees' Benefits 4 2012-04-01 2012-04-01 false Published information. 902.3 Section 902.3 Employees' Benefits JOINT BOARD FOR THE ENROLLMENT OF ACTUARIES RULES REGARDING AVAILABILITY OF INFORMATION § 902.3 Published information. (a) Federal Register. Pursuant to sections 552 and 553 of title 5 of the...

20 CFR 902.3 - Published information.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 20 Employees' Benefits 4 2013-04-01 2013-04-01 false Published information. 902.3 Section 902.3 Employees' Benefits JOINT BOARD FOR THE ENROLLMENT OF ACTUARIES RULES REGARDING AVAILABILITY OF INFORMATION § 902.3 Published information. (a) Federal Register. Pursuant to sections 552 and 553 of title 5 of the...

20 CFR 902.3 - Published information.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 20 Employees' Benefits 4 2014-04-01 2014-04-01 false Published information. 902.3 Section 902.3 Employees' Benefits JOINT BOARD FOR THE ENROLLMENT OF ACTUARIES RULES REGARDING AVAILABILITY OF INFORMATION § 902.3 Published information. (a) Federal Register. Pursuant to sections 552 and 553 of title 5 of the...

20 CFR 902.3 - Published information.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 20 Employees' Benefits 3 2011-04-01 2011-04-01 false Published information. 902.3 Section 902.3 Employees' Benefits JOINT BOARD FOR THE ENROLLMENT OF ACTUARIES RULES REGARDING AVAILABILITY OF INFORMATION § 902.3 Published information. (a) Federal Register. Pursuant to sections 552 and 553 of title 5 of the...

76 FR 55256 - Section 6707A and the Failure To Include on Any Return or Statement Any Information Required To...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-07

... Section 6707A and the Failure To Include on Any Return or Statement Any Information Required To Be... respect to the penalties applicable to the failure to include on any return or statement any information... (TD 9425) relating to the penalty under section 6707A for the failure to include on any return or...

Headache - Multiple Languages

MedlinePlus

... Section Headaches - العربية (Arabic) Bilingual PDF Health Information Translations Chinese, Simplified (Mandarin dialect) (简体中文) Expand Section Headaches - ... Chinese, Simplified (Mandarin dialect)) Bilingual PDF ... Health Information Translations Chinese, Traditional (Cantonese dialect) (繁體中文) Expand Section Headaches - ...