Requirements for the selective degradation of CD4 receptor molecules by the human immunodeficiency virus type 1 Vpu protein in the endoplasmic reticulum.

BACKGROUND: HIV-1 Vpu targets newly synthesized CD4 receptor for rapid degradation by a process reminiscent of endoplasmic reticulum (ER)-associated protein degradation (ERAD). Vpu is thought to act as an adaptor protein, connecting CD4 to the ubiquitin (Ub)-proteasome degradative system through an interaction with beta-TrCP, a component of the SCFbeta-TrCP E3 Ub ligase complex. RESULTS: Here, we provide direct evidence indicating that Vpu promotes trans-ubiquitination of CD4 through recruitment of SCFbeta-TrCP in human cells. To examine whether Ub conjugation occurs on the cytosolic tail of CD4, we substituted all four Ub acceptor lysine residues for arginines. Replacement of cytosolic lysine residues reduced but did not prevent Vpu-mediated CD4 degradation and ubiquitination, suggesting that Vpu-mediated CD4 degradation is not entirely dependent on the ubiquitination of cytosolic lysines and as such might also involve ubiquitination of other sites. Cell fractionation studies revealed that Vpu enhanced the levels of ubiquitinated forms of CD4 detected in association with not only the ER membrane but also the cytosol. Interestingly, significant amounts of membrane-associated ubiquitinated CD4 appeared to be fully dislocated since they could be recovered following sodium carbonate salt treatment. Finally, expression of a transdominant negative mutant of the AAA ATPase Cdc48/p97 involved in the extraction of ERAD substrates from the ER membrane inhibited Vpu-mediated CD4 degradation. CONCLUSION: Taken together, these results are consistent with a model whereby HIV-1 Vpu targets CD4 for degradation by an ERAD-like process involving most likely poly-ubiquitination of the CD4 cytosolic tail by SCFbeta-TrCP prior to dislocation of receptor molecules across the ER membrane by a process that depends on the AAA ATPase Cdc48/p97.

Selective alterations of brain osmolytes in acute liver failure: protective effect of mild hypothermia.

The principal cause of mortality in patients with acute liver failure (ALF) is brain herniation resulting from intracranial hypertension caused by a progressive increase of brain water. In the present study, ex vivo high-resolution 1H-NMR spectroscopy was used to investigate the effects of ALF, with or without superimposed hypothermia, on brain organic osmolyte concentrations in relation to the severity of encephalopathy and brain edema in rats with ALF due to hepatic devascularization. In normothermic ALF rats, glutamine concentrations in frontal cortex increased more than fourfold at precoma stages, i.e. prior to the onset of severe encephalopathy, but showed no further increase at coma stages. In parallel with glutamine accumulation, the brain organic osmolytes myo-inositol and taurine were significantly decreased in frontal cortex to 63\% and 67\% of control values, respectively, at precoma stages (p<0.01), and to 58\% and 67\%, respectively, at coma stages of encephalopathy (p<0.01). Hypothermia, which prevented brain edema and encephalopathy in ALF rats, significantly attenuated the depletion of myo-inositol and taurine. Brain glutamine concentrations, on the other hand, did not respond to hypothermia. These findings demonstrate that experimental ALF results in selective changes in brain organic osmolytes as a function of the degree of encephalopathy which are associated with brain edema, and provides a further rationale for the continued use of hypothermia in the management of this condition.

Variations interindividuelles des performances cognitives et conséquences évolutives chez une population naturelle de mésange charbonnière (Parus major)

Les animaux font face à des changements environnementaux brutaux dus aux modifications de milieux liés à l'activité humaine et aux changements climatiques, et doivent s'ajuster rapidement à leur nouvel environnement. Certains processus cognitifs comme l'innovation et l'apprentissage permettent aux animaux d'intégrer de nouveaux comportements à leur répertoire comportemental (flexibilité comportementale), leur donnant l'opportunité d'intégrer un comportement plus optimal pour s'ajuster. Les performances cognitives varient entre espèces et les individus d'une même population et bien que des études récentes se soient intéressées aux causes des variations interindividuelles des performances cognitives, les conséquences restent peu explorées. Dans cette thèse, les questions des pressions de sélection s'exerçant sur les capacités cognitives sont abordées afin de mieux comprendre l'évolution de ces traits au sein d'une population naturelle de mésange charbonnière Parus major. Un nouveau test de résolution de problème a tout d'abord été présenté à des couples reproducteurs directement en milieu naturel. Les résultats ont montré que les couples les plus performants à résoudre la tâche surpassaient les couples les moins performants sur plusieurs mesures de succès reproducteur. Afin de vérifier que la motivation à nourrir les poussins ne biaisait pas cette relation, la taille de nichée a ensuite été manipulée, ce qui n'a pas affecté la performance subséquente des parents. Les couples innovateurs démontraient un meilleur succès reproducteur quel que soit le changement de la taille de nichée subit, ce qui suggère que cette performance influence bien le succès de reproduction, et non l'inverse. De plus, les couples innovateurs approvisionnaient leurs poussins plus souvent que les couples non innovateurs, suggérant que les innovateurs pourraient exploiter leur habitat de façon plus optimale. Dans un troisième temps, plusieurs caractéristiques morphologiques, dont la coloration des plumes, ont été reliées aux performances de résolution de problème et d'apprentissage. Ces liens, bien que complexes et condition-dépendants, pourraient indiquer un rôle de ces performances lors de la sélection sexuelle. Enfin, afin de tester l'effet du parasite sanguin du paludisme sur les traits comportementaux, un médicament contre le paludisme a été injecté à des femelles reproductrices. Cette injection n'a pas modifié leurs performances cognitives mais a augmenté leur niveau d'activité et d'exploration du nichoir en réponse à la tâche de résolution de problème. Ce parasite sanguin, très présent chez les populations depassereaux, pourrait donc expliquer les variations interindividuelles et interpopulationnelles de certains traits comportementaux en milieu naturel, au même titre que dans nombreux autres systèmes hôte-parasites étudiés. Les travaux présentés dans cette thèse ont permis de détailler pour la première fois la relation entre une performance cognitive et le succès reproducteur chez une population aviaire naturelle, une relation robuste et non influencée par la motivation à nourrir la couvée. Cette performance cognitive est reliée à plusieurs traits morphologiques, mais non à la charge parasitaire. Une meilleure exploitation de l'habitat et habileté à s'occuper des poussins pourrait expliquer cette relation.

Rôle de la tolérance centrale et périphérique des lymphocytes T autoréactifs dans deux nouveaux modèles murins double transgéniques

Les maladies autoimmunes sont des affections chroniques, le plus souvent invalidantes, qui touchent plus de 5% de la population dans les pays développés. L’autoimmunité résulte de la rupture des mécanismes de tolérance du système immunitaire vis-à-vis des autoantigènes exprimés par les tissus de l’organisme, entraînant la destruction d’un ou de plusieurs organes-cibles par les lymphocytes T et/ou B. L’hépatite autoimmune et le diabète autoimmun se caractérisent par la destruction sélective des hépatocytes et des cellules beta pancréatiques, respectivement. De plus en plus d’arguments suggèrent une implication des lymphocytes T CD8+ dans le déclenchement, la progression et la régulation des réponses associées à plusieurs maladies autoimmunes. Dans ce projet, nous avons suivi l’évolution de clones de lymphocytes T CD8+ spécifiques à un antigène particulier dont le site d’expression différait. Pour ce faire, nous avons développé deux nouveaux modèles murins double transgéniques par croisement entre une lignée de souris exprimant un TCR transgénique spécifique à la nucléoprotéine (NP) du virus de la chorioméningite lymphocytaire (LCMV), et une souris exprimant cette NP-LCMV : 1) uniquement dans les hépatocytes (modèle d’hépatite autoimmune), ou 2) simultanément dans le thymus et le pancréas (modèle de diabète autoimmun). L’avidité fonctionnelle des lymphocytes T CD8+ spécifiques à la NP chez les souris TCR transgéniques était inversement proportionnelle au niveau d’expression du TCR. Le répertoire lymphocytaire dans le thymus, la rate, les ganglions et le sang périphérique a été caractérisé pour chacune des lignées de souris double transgéniques, de même que la capacité fonctionnelle et le phénotype (marqueurs d’activation/mémoire) des lymphocytes T CD8+ autoréactifs. Chacun des deux nouveaux modèles présentés dans cette étude ont montré que les lymphocytes T CD8+ spécifiques à la NP sont aptes à briser la tolérance centrale et périphérique et à provoquer une réaction d’autoimmunité spontanée. Dans le modèle d’hépatite autoimmune, où l’expression de l’autoantigène était restreinte au foie, la surexpression du TCR transgénique a entraîné une délétion thymique quasi-totale des lymphocytes T CD8+ spécifiques à la NP prévenant le développement d’une hépatite spontanée. alors qu’un niveau de TCR comparable à celui d’une souris de type sauvage a permis une sélection positive des lymphocytes autoréactifs qui se sont accumulés dans le foie où ils se sont activés pour provoquer une hépatite autoimmune spontanée. Dans le modèle de diabète autoimmun, où l’autoantigène était exprimé dans le pancréas et le thymus, les souris des deux lignées double transgéniques ont montré une délétion thymique partielle, peu importe le niveau d’expression du TCR. Seuls les mâles adultes développaient un diabète spontané et une partie de leurs lymphocytes T CD8+ exprimaient une combinaison particulière de marqueurs d’activation/mémoire (CD44, CD122, PD-1). Cette population lymphocytaire était absente chez les souris femelles et les mâles sains. L’étude de la tolérance des lymphocytes T CD8+ autoréactifs dans nos deux nouveaux modèles murins double transgéniques a permis d’identifier des mécanismes alternatifs possiblement impliqués dans la tolérance et l’activation, et de mieux comprendre le rôle des lymphocytes T CD8+ autoréactifs dans le processus autoimmun menant à l’hépatite autoimmune et au diabète autoimmun. Ces découvertes seront utiles pour développer de nouvelles approches thérapeutiques ciblant les lymphocytes T CD8+ autoréactifs.

Selective inhibition of inducible nitric oxide synthase prevents lipid peroxidation in cartilage from patients with osteoarthritis.

INTRODUCTION: Emerging evidence indicates that nitric oxide (NO), which is increased in osteoarthritic (OA) cartilage, plays a role in 4-hydroxynonenal (HNE) generation through peroxynitrite formation. HNE is considered as the most reactive product of lipid peroxidation (LPO). We have previously reported that HNE levels in synovial fluids are more elevated in knees of OA patients compared to healthy individuals. We also demonstrated that HNE induces a panoply of inflammatory and catabolic mediators known for their implication in OA cartilage degradation. The aim of the present study was to investigate the ability of inducible NO synthase (iNOS) inhibitor, L-NIL (L-N6-(L-Iminoethyl)Lysine), to prevent HNE generation through NO inhibition in human OA chondrocytes. METHOD: Cells and cartilage explants were treated with or without either an NO generator (SIN or interleukin 1beta (IL-1β)) or HNE in absence or presence of L-NIL. Protein expression of both iNOS and free-radical-generating NOX subunit p47 (phox) were investigated by western blot. iNOS mRNA detection was measured by real-time RT-PCR. HNE production was analysed by ELISA, Western blot and immunohistochemistry. S-nitrosylated proteins were evaluated by Western Blot. Prostaglandin E2 (PGE2) and metalloproteinase 13 (MMP-13) levels as well as glutathione S-transferase (GST) activity were each assessed with commercial kits. NO release was determined using improved Griess method. Reactive oxygen species (ROS) generation was revealed using fluorescent microscopy with the use of commercial kits. RESULTS: L-NIL prevented IL-1β-induced NO release, iNOS expression at protein and mRNA levels, S-nitrosylated proteins and HNE in a dose dependent manner after 24h of incubation. Interestingly, we revealed that L-NIL abolished IL-1β-induced NOX component p47phox as well as ROS release. The HNE-induced PGE2 release and both cyclooxygenase-2 (COX-2) and MMP-13 expression were significantly reduced by L-NIL addition. Furthermore, L-NIL blocked the IL-1β induced inactivation of GST, an HNE-metabolizing enzyme. Also, L-NIL prevented HNE induced cell death at cytotoxic levels. CONCLUSION: Altogether, our findings support a beneficial effect of L-NIL in OA by preventing LPO process in NO-dependent and/or independent mechanisms.

Studies on Soft Bycatch Reduction Devices for Selective Trawling

Trawling, though an efficient method of fishing, is known to be one of the most non-selective methods of fish capture. The bulk of the wild caught penaeid shrimps landed in India are caught by trawling.In addition to shrimps, the trawler fleet also catches considerable amount of non-shrimp resources. The term bycatch means that portion of the catch other than target species caught while fishing, which are either retained or discarded. Bycatch discards is a serious problem leading to the depletion of the resources and negative impacts on biodiversity. In order to minimize this problem, trawling has to be made more selective by incorporating Bycatch Reduction Devices (BRDs). There are several advantages in using BRDs in shrimp trawling. BRDs reduce the negative impacts of shrimp trawling on marine community. Fishers could benefit economically from higher catch value due to improved catch quality, shorter sorting time, lower fuel costs, and longer tow duration. Adoption of BRDs by fishers would forestall criticism by conservation groups against trawling.

A Chloride Ion-Selective Potentiometric Sensor Based on a Polymeric Schiff Base Complex

This paper describes the fabrication of an ion-selective electrode in which a polymeric Schiff base complex of cobalt(II) is used as the ionophore.The main advantage of the electrode is that it is mechanically stable upto 3 months..The electrode shows a linear response in the range of 2.5 × 10-5-0.5 × 10-1 mol dm-3. The response time of the electrode is 30 s.The pH range at which the electrode works is 3.8 to 6.8. The electrode was found to be selective towards chloride ion in the presence of ions like Na+, Ca2+, Mn2+, ,Fe3+, Co2+, Ni2+, Cu2+, Zn2+, CH3COO-, NO3-, SO42- ,Br- and NO2-.

Investigations on Hard Bycatch Reduction Devices for Selective Trawling

In the context of Indian fisheries there is a paucity of information on bycatch, in general, and bycatch reduction technologies, in particular. In this study, a detailed investigation on trawl bycatch and bycatch reduction measures is attempted with a view to evolve optimized BRDs for improving selectivity of commercial shrimp trawls. The objectives of the study included design and development of hard bycatch reduction devices (BRDs), comparative evaluation of hard bycatch reduction devices, for selective trawling, bycatch characterisation of the trawl landings, off Central Kerala; and investigations on status of the existing trawling systems operated off Central Kerala.

Fabrication of Potentiometric Sensors for the Selective Determination of Ketoconazole

The fabrication and analytical applications of two types of potentiometric sensors for the determination of ketoconazole (KET) are described. The sensors are based on the use of KET-molybdophosphoric acid (MPA) ion pair as electroactive material. The fabricated sensors include both polymer membrane and carbon paste electrodes. Both sensors showed a linear, stable and near Nernstian slope of 57.8mV=decade and 55.2mV=decade for PVC membrane and carbon paste sensors respectively over a relatively wide range of KET concentration (1×10-2-5×10-5 and 1×10-2-1×10-6). The sensors showed a fast response time of <30 sec and <45 sec. A useful pH range of 3–6 was obtained for both types of sensors. A detection limit of 2.96 10 5M was obtained for PVC membrane sensor and 6.91 10 6M was obtained for carbon paste sensor. The proposed sensors proved to have a good selectivity for KET with respect to a large number of ions. The proposed sensors were successfully applied for the determination of KET in pharmaceutical formulations. The results obtained are in good agreement with the values obtained by the standard method.

Selective N-monomethylation of aniline over Zn1-xNixFe2O4 (x=0, 0.2, 0.5, 0.8 and 1) type systems

The reaction of aniline with methanol was carried out over Zn1-xNixFe2O4 (x= 0, 0.2, 0.5, 0.8 and 1) type systems in a fixed-bed down-flow reactor. It was observed that systems possessing low ``x'' values are highly selective and active for mono N-alkylation of aniline leading to N-methyl aniline. Selectivity for N-methyl aniline over ZnFe2O4 was more than 99% under the optimized reaction conditions. Even at methanol to aniline molar ratio of 2, the yield of N-methyl aniline was nearly 55.5%, whereas its yield exceeded 67% at the molar ratio of 7. The Lewis acid sites of the catalysts are mainly responsible for the good catalytic performance. Cation distribution in the spinel lattice influences their acido-basic properties, and hence, these factors have been considered as helpful to evaluate the activity and stability of the systems.

Selective N-monomethylation of aniline using Zn1-x CoxFe2O4( x=0, 0.2, 0.5, 0.8 and 1.0)type systems

A series of ferrites having the general formula Zn1-xCoxFe2O4 (x=0, 0.2, 0.5, 0.8 and 1.0)were prepared by soft chemical route. The materials were characterized by adopting various physico-chemical methods. The reaction of aniline with methanol was studied in a fixed-bed reactor system as a potential source for the production of various methyl anilines. It was observed that systems possessing low ‘ x’ values are highly selective and active for N-monoalkylation of aniline leading to N-methylaniline. Reaction parameters were properly varied to optimize the reaction conditions for obtaining N-methylaniline selectively and in better yield. Among the systems Zn0.8Co0.2Fe2O4 is remarkable due to its very high activity and excellent stability. Under the optimized conditions N-methylaniline selectivity exceeded 98%. Even at a methanol to aniline molar ratio of 2, the yield of N-methylaniline was nearly 50%, whereas its yield exceeded 71% at the molar ratio of 5. ZnFe2O4, though executed better conversion than Zn0.8Co0.2Fe2O4 in the initial period of the run, deactivates quickly as the reaction proceeds. The Lewis acidity of the catalysts is mainly responsible for the good performance. Cation distribution in the spinel lattice influences their acido-basic properties and, hence, these factors have been considered as helpful parameters to evaluate the activity of the systems.

Selective Methyl Tert-Butyl Ether Synthesis over Metal Exchanged Pillared Clays

Iron and mixed iron aluminium pillared montmorillonites prepared by partial hydrolysis method was subjected to room temperature exchange with transition metals of the first series. The materials exhibit good structural as well as thermal stability. Exchanged metals were found to be present inside the porous network, in the environs of the pillars. Mixed pillaring resulted in the intercalation of Al 13 like polymers in which Al is partially substituted by Fe. The acidic structure was followed by temperature programmed desorption of ammonia and cumene cracking test reaction. Weak and medium sites overshadow the strong sites in all systems. However, exchange with metals increases the number of strong sites. The prepared materials are efficient catalysts for gas phase MTBE synthesis. The catalytic activity can be well correlated with the total amount of weak and medium acid sites.

Selective alkylation of aniline to N-methyl aniline using chromium manganese ferrospinels

Various compositions of chromium manganese ferrospinels were tested as catalysts for the vapour phase alkylation of aniline with methanol. The samples were prepared by room temperature co-precipitation technique and characterized by various physico-chemical methods. The acidity–basicity determination revealed that the samples possess greater amount of basic sites than acidic sites. All the ferrite samples proved to be selective and active for N-monoalkylation of aniline leading to N-methyl aniline; Cr0.6Mn0.4Fe2O4, Cr0.8Mn0.2Fe2O4 and CrFe2O4 exhibited cent percent selectivity for N-methyl aniline. Neither C-alkylated products nor any other side products were detected for all catalyst samples. The catalytic activity of the samples studied in this reaction is related to their acid–base properties and also on the cation distribution. Under the optimized reaction conditions all the systems showed constant activity for a long duration.

Selective mode excitation in dye-doped DNA polyvinyl alcohol thin film

A solid-state laser based on a dye-doped deoxyribonucleic acid (DNA) matrix is described. A thin solid film of DNA has been fabricated by treating with polyvinyl alcohol (PVA) and used as a host for the laser dye Rhodamine 6G. The edge emitted spectrum clearly indicated the existence of laser modes and amplified spontaneous emission. Lasing was obtained by pumping with a frequency-doubled Nd:YAG laser at 532 nm. For a pump energy of 10 mJ/pulse, an intense line with FWHM ≈0.2 nm was observed at 566 nm due to selective mode excitation.