In Silico Structural Studies and Molecular Docking Analysis of Delta6-desaturase in HUFA Biosynthetic Pathway.

Fish are an important source of highly unsaturated fatty acids (HUFA) such as eicosapentaenoic acid EPA (20:5 n-3) and docosahexaenoic acid DHA (22:6 n-3) and play a significant role in human nutrition. The fatty acyl delta6-desaturase (Δ6 desaturase) is a rate-limiting enzyme in the biosynthetic pathway of highly unsaturated fatty acids (HUFA) that converts polyunsaturated fatty acids (PUFA) such as linoleic (18:2n-6) and α-linolenic (18:3n-3) acids into HUFA. In this study, fatty acyl Δ6 desaturase was identified from pangasius (Pangasianodon hypophthalmus) and further analyzed for sequenced-based characterization and 3D structural conformation. Sequenced-based analysis revealed some important secondary information such as physicochemical property. e.g., isoelectric point, extinction coefficient, aliphatic index, and grand average hydropathy, among others, and also post-translational modification sites were identified. An evolutionary-conserved stretch of amino acid residue and a functionally significant conserved structural ancestor, N-terminal cytochrome b5 and membrane FADS-like superfamily, were identified. Protein association analysis showed a high confidence score with acyl-CoA synthetase, elovl5, elovl2, and phospholipase A2. Herein, we report, for the first time, a 3D native structure of Δ6 desaturase protein by homology modeling approach; molecular docking analysis was performed with linoleic (18:2n-6) and α-linolenic (18:3n-3) acids, which are the two key substrates in the HUFA biosynthetic pathway. This work provides insight into the structural and functional characterization of Δ6 desaturase, which is involved in HUFA biosynthesis as a rate-limiting enzyme.

Emissivity range constraints algorithm for multi-wavelength pyrometer (MWP).

In order to realize rapid and real temperature measurement for high temperature targets by multi-wavelength pyrometer (MWP), emissivity range constraints to optimize data processing algorithm without effect from emissivity has been developed. Through exploring the relation between emissivity deviation and true temperature by fitting of large number of data from different emissivity distribution target models, the effective search range of emissivity for every time iteration is obtained, so data processing time is greatly reduced. Simulation and experimental results indicate that calculation time is less by 0.2 seconds with 25K absolute error at 1800K true temperature, and the efficiency is improved by more than 90% compared with the previous algorithm. The method has advantages of simplicity, rapidity, and suitability for in-line high temperature measurement.

Dietary lecithin potentiates thermal tolerance and cellular stress protection of milk fish (Chanos Chanos) reared under low dose endosulfan-induced stress.

Endosulfan is an organochlorine pesticide commonly found in aquatic environments that has been found to reduce thermal tolerance of fish. Lipotropes such as the food additive, Lecithin has been shown to improve thermal tolerance in fish species. This study was conducted to evaluate the role of lipotropes (lecithin) for enhancing the thermal tolerance of Chanos chanos reared under sublethal low dose endosulfan-induced stress. Two hundred and twenty-five fish were distributed randomly into five treatments, each with three replicates. Four isocaloric and isonitrogenous diets were prepared with graded levels of lecithin: normal water and fed with control diet (En0/L0), endosulfan-treated water and fed with control diet (En/L0), endosulfan-treated water and fed with 1% (En/L1%), 1.5% (En/L 1.5%) and 2% (En/L 2%) lecithin supplemented feed. The endosulfan in treated water was maintained at the level of 1/40th of LC50 (0.52ppb). At the end of the five weeks, critical temperature maxima (CTmax), lethal temperature maxima (LTmax), critical temperature minima (CTmin) and lethal temperature minima (LTmin) were Determined. There was a significant (P<0.01) effect of dietary lecithin on temperature tolerance (CTmax, LTmax, CTmin and LTmin) of the groups fed with 1, 1.5 and 2% lecithin-supplemented diet compared to control and endosulfan-exposed groups. Positive correlations were observed between CT max and LTmax (R(2)=0.934) as well as between CTmin and LTmin (R(2)=0.9313). At the end of the thermal tolerance study, endosulfan-induced changes in cellular stress enzymes (Catalase, SOD and GST in liver and gill and neurotansmitter enzyme, brain AChE) were significantly (p<0.01) improved by dietary lecithin. We herein report the role of lecithin in enhancing the thermal tolerance and protection against cellular stress in fish exposed to an organochlorine pesticide.

Evaluation of pectinolytic activities for oenological uses from psychrotrophic yeasts.

UNLABELLED: Of the twenty-three morphotypes of yeasts isolated from soil capable of utilizing pectin as sole carbon source at 6°C, two yeast isolates, one psychrotolerant (PT1) and one psychrophilic (SPY11), were selected according to their ability to secrete pectinolytic enzymes under some oenological conditions (temperature 6 and 12°C and pH 3.5) and ability or inability to grow above 20°C, respectively. As compared to their optimal activity, the three pectinolytic enzymes viz., pectin methyl esterase (PME), endopolygalacturonase (endo-PG) and exopolygalacturonase (exo-PG) isolated and assayed at pH 3.5 from PT1 were found to retain 39, 60 and 60% activity at 12°C and 40, 79 and 74% activity at 28°C, respectively. Likewise, the enzymes PME and endo-PG at pH 3.5 from SPY11 displayed 46 and 86% activity at 12°C and 50 and 60% activity at 28°C, respectively. All these enzymes showed 20-90% of residual activity at pH 3.5 and 6°C. The yeast isolates PT1 and SPY11 were identified as Rhodotorula mucilaginosa and Cystofilobasidium capitatum, respectively, on the basis of morphological, physiological and molecular characteristics. This study presents the first report on pectinolytic activities under major oenological conditions from psychrotolerant isolate R. mucilaginosa PT1 and psychrophilic isolate C. capitatum SPY11. SIGNIFICANCE AND IMPACT OF THE STUDY: The cold-active pectinolytic enzymes (PME, endo-PG and exo-PG) from the newly isolated and identified psychrophilic yeast Cystofilobasidium capitatum SPY11 and psychrotolerant yeast Rhodotorula mucilaginosa PT1that exhibited 50-80% of their optimum activity under some major oenological conditions pH (3.5) and temperatures (6 and 12°C) could be applied to wine production and juice clarification at low temperature. The psychrotrophic yeasts themselves could be applied to cold process for the production of enzymes thus saving cost of energy and protecting process from contamination.

Methyl donors potentiates growth, metabolic status and neurotransmitter enzyme in Labeo rohita fingerlings exposed to endosulfan and temperature.

A 2-month preliminary study was conducted to delineate the effect of dietary methyl donors (choline, betaine, and lecithin) on the growth performance and metabolic status of Labeo rohita fingerlings subjected to endosulfan alone and in combination with elevated temperature. Four iso-caloric and iso-nitrogenous diets viz. basal diet, betaine-supplemented diet, choline-supplemented diet and lecithin-supplemented diet were prepared and fed to the different experimental groups throughout the experimental period as per the design. Two hundred and seventy fingerlings (average weight 7.95 ± 0.04 g) were randomly distributed in six treatment groups each having three replicates. The experimental groups were as follows: fish subjected to normal water (without endosulfan) and fed with control diet (control group T(0)), fish subjected to endosulfan-treated water and fed with control diet (T(1)), fish subjected to concurrent exposure of endosulfan and elevated temperature and fed with control diet (T(2)), fish subjected to endosulfan and elevated temperature and fed with choline-supplemented diet (T(3)), fish subjected to endosulfan and temperature and fed with betaine-supplemented feed (T(4)), and fish subjected to endosulfan and temperature and fed with lecithin-supplemented feed (T(5)). The result shows that in both the groups, that is, endosulfan exposed and concurrent exposure to endosulfan and elevated temperature group of L. rohita the growth performance like percentage weight gain, feed conversion ratio and specific growth rates were significantly different (P < 0.01) when fed with supplemented diet compared with control fed group. The liver LDH and MDH activity were significantly lower in lecithin, betaine, and choline fed groups. The muscle AST as well as G6PDH, AST, and ALT did not vary but liver ALT, gill and liver ATPase, intestine ALP, muscle and liver glycogen varied significantly with dietary supplementation. The liver and gill glutathione-S-transferase (GST) activities were significantly lower in methyl donors-supplemented groups and brain AChE activity showed lower inhibition in supplemented groups in both endosulfan alone and concurrently exposed endosulfan and temperature groups. The result obtained in this study concludes that inclusion of methyl donors, particularly lecithin and betaine in feed as nutritional supplements have potential to improve growth and stress mitigating effect in L. rohita fingerlings.

Dietary choline, betaine and lecithin mitigates endosulfan-induced stress in Labeo rohita fingerlings.

A five-week experiment was conducted to delineate stress-mitigating effects of three different methyl donors in Labeo rohita fingerlings subjected to endosulfan toxicity. Four iso-nitrogenous and iso-caloric feed were prepared with and without supplementation of methyl donors. The feed were basal or control diet (i.e., without methyl donor supplementation), feed supplemented with choline, feed supplemented with betaine and feed supplemented with lecithin. Two hundred and twenty-five fishes were distributed randomly in five treatment groups each with three replicates. The experimental setup were normal water (without endosulfan) and fed with control diet (control group), endosulfan-treated water and fed with control diet (T1), endosulfan-treated water and fed with choline supplemented feed (T2), endosulfan-treated water and fed with betaine supplemented feed (T3) and endosulfan-treated water and fed with lecithin-supplemented feed (T4). The level of endosulfan in endosulfan treated water was maintained at the level of 1/10 of LC50, that is, 0.2 ppb. During the experiment, growth performances, metabolic enzyme activity and histological examination were done to assess the effect of treatments. The growth performance (percentage weight gain, feed conversion ratio, specific growth rate and protein efficiency ratio) and nutrient digestibility were significantly different (P<0.01) in lecithin, betaine and choline fed group when compared to endosulfan-exposed group fed with basal diet. The liver LDH and MDH activity were significantly (P<0.01) improved in the groups fed with methyl donor supplemented diet. The liver AST and ALT, brain AChE and muscle ALT did not change with supplementation in the diet, but muscle ALT and G6PDH significantly (P<0.01) changed with supplementation. The gill and liver ATPase and intestinal ALP were significantly (P<0.01) noticeably changed in supplemented group. After endosulfan exposure, histopathology alter like slight large vacuolation in hepatocyte and lipoid vacuole were observed and with supplementation normal appearance of liver were observed. The chromosome aberration (karyotype) was observed in endosulfan-exposed group. The result obtained in present study concluded that inclusion of methyl donors, particularly lecithin and betaine, in feed as nutritional supplements has a potential stress-mitigating effect in L. rohita fingerlings.

Theoretical and experimental performance evaluation of single-slope single-basin solar still with multiple V-shaped floating wicks.

A solar still is used to convert saline water into potable water by means of the distillation process. In order to improve the productivity of conventional solar still, various modifications are implemented by researchers. In the present study, multiple V-shaped floating wicks are used to enhance heat absorption and thereby increase productivity. The experiments are performed during the summer and winter seasons in Rewa, India (Latitude: 24.5373° N; Longitude: 81.3042° E). These multiple floating wicks are made from black jute cloth wrapped in V-shaped pieces of thermocol. Because of their V-shaped profile, the evaporative surface area of modified solar still is 26% larger than that of conventional solar still. The maximum daily productivity in one of the clear days is found to be approximately 6.20 kg/m2 in summer and 3.23 kg/m2 in winter with daily efficiencies of 56.62% and 47.75%, respectively. A theoretical thermal model is formulated by using the energy balance equations of the modified solar still. Reasonable agreement was seen between the theoretical and experimental results of modified solar still. An economic analysis is also performed for the modified solar still and conventional solar still; in a 10-year life cycle, the annual cost of distilled water is estimated at Rs. 1.81/kg for the former and Rs. 2.24/kg for the latter.

Early Eocene lagomorph (Mammalia) from Western India and the early diversification of Lagomorpha.

We report the oldest known record of Lagomorpha, based on distinctive, small ankle bones (calcaneus and talus) from Early Eocene deposits (Middle Ypresian equivalent, ca 53 Myr ago) of Gujarat, west-central India. The fossils predate the oldest previously known crown lagomorphs by several million years and extend the record of lagomorphs on the Indian subcontinent by 35 Myr. The bones show a mosaic of derived cursorial adaptations found in gracile Leporidae (rabbits and hares) and primitive traits characteristic of extant Ochotonidae (pikas) and more robust leporids. Together with gracile and robust calcanei from the Middle Eocene of Shanghuang, China, also reported here, the Indian fossils suggest that diversification within crown Lagomorpha and possibly divergence of the family Leporidae were already underway in the Early Eocene.

Antioxidants prolong life span and inhibit the senescence-dependent accumulation of fluorescent pigment (lipofuscin) in clones, of Podospora anserina.

Culture of clones of Podospora anserina s+ with either nordihydroguaiaretic acid or reduced glutathione (GSH) at concentrations that were not inhibitory to growth significantly prolonged the average time to onset of senscence. GSH also prolonged the average time to onset of clonal death. The specific concentration of chloroform-methanol soluble fluorescent pigment was larger in senescent than in pre-senescent cells. The pigment exhibited fluorescence excitation and emission spectra and fluorescence polarization numbers characteristic of lipofuscin, an end-product of lipid peroxidation. Analyses of the lipofuscin concentration in either sub-clonal fractions of different times of origin from senescent clones, or in sub-clonal fractions of identical age in time of origin from parent clones of different age, revealed a similar concentrations distribution. Although pre-senscent cells contained rather large concentrations, a massive increase occurred during senescence prior to the time of onset of clonal death. Culture with GSH not only prolonged clonal life span but aslo inhibited the formation of lipofuscin by an average factor of 30. Furthermore, unlike untreated clones, the sub-clonal distribution of the pigment was not only low but was also independent of their age.

Ageing of Neurospora crassa. VII. Accumulation of fluorescent pigment (lipofuscin) and inhibition of the accumulation by nordihydroguairetic acid.

Continuous administration of the antioxidant nordihydroguairetic acid to clones of the natural-death nutant or wild-type Neurospora crassa growing on 5-fluorotryptophan not only alleviates the time-dependent deterioration of growth rate (senescence), but also inhibits the accumulation of a fluorescent pigment (lipofuscin), an end-product of lipid peroxidation. In the three model system of ageing in N. crassa that we have examined, it is now clear that massive accumulation of lipofuscin begins shortly before or concident with the period of senescence. By this and other criteria, the youngest cells in time of origin appear to be most senescent. Thus the degree of senscence appears to be a function of physiological rather than chronological age and, more specifically, is probably a function of the number of nuclear mitotic divisions. Normal, non-senescent, wild-type, clones also accumulate lipofuscin; but, unlike clones undergoing senscence, accumulate very low concentrations which are independent of subclonal age. Apparently, clones of wild type can tolerate a low level of lipid peroxidation without undergoing senescence; but even a low level is inimical since culture with nordihydroguaiaretic acid not only enhances their growth rate, but also inhibits the accumulation of lipofuscin.

Genetically determined conidial longevity is positively correlated with superoxide dismutase, catalase, glutathione peroxidase, cytochrome c peroxidase, and ascorbate free radical reductase activities in Neurospora crassa.

Aging of post-mitotic cells, the conidia, of Neurospora crassa is defined as the time-dependent loss of viability under a constant laboratory environment which probably resembles the organism's tropical habitat; namely, at 30 degrees C, 85-100% relative humidity under white light. Median lifespan is defined as the age at which survival of a conidial population has declined to 50% of that of a fully viable population at birth. A collection of short (age-) and long-lived (age+) mutants were previously selected from the wild-type whose median lifespan is 22 days. Thus, five groups of strains with distinct lifespans of 7, 22, 36, 50 and 60 days were defined. The purposes of the present investigation were to determine if the activities of anti-oxygenic enzymes are correlated with lifespan and to elucidate the function of the cellular longevity determinant genes. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) were highly-correlated with lifespan; whereas glutathione reductase and non-specific peroxidase activities were not correlated. The short-lived mutants were also deficient in cytochrome c peroxidase (CPX) and ascorbate free radical reductase (AFR), but not deficient in dehydroascorbate reductase. (These latter three enzymes were not examined in age+ mutants.) By isoelectric focusing analysis, the deficiencies of SOD, CAT, and GPX activities of age- mutants were defined in terms of specific isozymes. The mutants were specifically deficient in a cyanide-resistant mitochondrial isozyme of SOD. Sixteen age- genes, called the age-1 complex, were previously mapped on one arm of the seven chromosomes. On the basis of mapping and complementation data, it was inferred that the genes are spatially and functionally redundant. The hypothesis of functional redundancy is also supported by the enzyme data. Of seven mutants examined, representing seven of the age- genes, all were deficient in SOD, CAT and CPX, and six were deficient in AFR. Of four mutants examined, representing four of the genes, all were deficient in GPX. The results indicate a molecular basis for the previously observed photosensitivity of the mutants.(ABSTRACT TRUNCATED AT 400 WORDS)

Evidence for a defective mitochondrial membrane in 1,2-dimethylhydrazine-induced colon adenocarcinoma in rat: enhanced lipid peroxidation potential in vitro.

Enhanced lipid peroxidation potential was measured in Holtzman rat colon tumors induced by chronic subcutaneous injection of 1,2-dimethyl-hydrazine as compared with normal colonic tissue. The peroxidation potentials were determined in the mitochondrial cellular components by measuring the ferrous-ascorbate induced formation of malondialdehyde. The tumor mitochondria were found to peroxidize at a rate 8-10-fold higher than the comparable normal tissue components. In addition, we found that the mitochondria from the cancer cells exhibited reduced NADH-cytochrome c reductase activity. These observations suggest an involvement of non-enzymatic free radical flux in DMH-induced carcinogenesis, which may be the result of structurally altered mitochondrial membranes.

Vasopressin and oxytocin but not glucose stimulate hydrolysis of phosphatidylinositol and phosphatidylcholine in a hamster insulinoma.

HIT-T15 cells prelabeled with [3H]-arachidonate were incubated for 15 minutes at 37 degrees C in Krebs Ringer buffer (pH 7.1) in the presence and absence of various agonists. Radioactivity remaining in major phospholipids was measured at the end of incubation period. Oxytocin (1 microM), vasopressin (1 microM), and A23187 (5 microM) stimulated loss of radioactivity from phosphatidylinositol and phosphatidylcholine. No loss of radioactivity from either of the phospholipids, however, was detected in the presence of 10 mM D-glucose, an insulin secretagogue in HIT-T15 cells. The lack of phosphatidylinositol response to glucose was also evident when the cells were prelabeled with myo-[3H] inositol. The formation of inositol phosphates at 15 minutes was readily observed upon the treatment of myo-[3H] inositol-labeled cells with oxytocin or vasopressin but not glucose or A23187. Inability of glucose to stimulate phosphatidylinositol and phosphatidylcholine hydrolysis in beta cell-derived HIT-T15 cells contrasts sharply with results from studies with pancreatic islets, where hydrolysis of these two phospholipids is readily observed and thought to contribute to the signaling mechanism responsible for stimulation of insulin secretion.

Stimulation of phosphoinositide hydrolysis by glyceraldehyde-3-phosphate in electroporated HIT-T15 cells.

HIT-T15 cells labeled with myo-[3H] inositol were permeabilized by electroporation and subsequently stimulated with various glycolytic intermediates in the presence of 20 mM LiCl in a buffer mimicking cytosolic ionic composition. Of the various glycolytic intermediates tested, only D-glyceraldehyde-3-phosphate (G3-P) stimulated the formation of labeled inositol phosphates. The half-maximal response to G3-P occurred at a concentration of 0.75 mM. Formation of inositol phosphates in electroporated cells was also observed in response to GTP. G3-P further potentiated the formation of inositol phosphates in response to GTP, however, the interaction between G3-P and GTP was additive rather than synergistic, indicating that G3-P stimulates phosphoinositide hydrolysis in a manner different than the receptor mediated GTP-dependent activation of phospholipase C. The potentiation of the GTP response by G3-P did not appear to involve inhibition of the GTPase activity of a phosphoinositide-specific G protein, since G3-P also potentiated the formation of inositol phosphates in response to GTP-gamma-S or NaF in a nearly additive manner.

Inhibition of germination of Bacillus cereus T spores by phenylglyoxal.

Phenylgloxal at a concentration of 0.6 mM inhibited germination of Bacillus cereus T spores as characterized by a decrease in absorbance, dipicolinic acid and loss in heat resistance in a chemically defined growth and sporulation medium. In a germination medium containing L-alanine and adenosine, phenylglyoxal inhibited decrease in absorbance and affected partial loss of viability. It is postulated that phenylglyoxal interacts with free amino groups of various enzymes or amino compounds present in the spore structure thereby causing the inhibition of germination.

Radiation equivalency of 1, 2-dimethylhydrazine in rats.

A scale based upon the induction of anti-tumor cellular immune responses has been described which relates the in vivo mutagenic/carcinogenic potential of an environmental agent with that of ionizing radiation. Such a potential (radiation equivalency) has now been determined in rats for the gastrointestinal carcinogen 1, 2,-dimethylhydrazine as compared with the effects induced by X-irradiation of only the small intestine. The preliminary results suggest that such scales may be readily constructed based upon environmental immunology concepts which relate the potential hazard of an agent to be weighed alongside the maximum allowable population limits of ionizing radiation.

Breast feeding and weaning practices among urban Muslims of district Lucknow.

PIP: Data on 73 Moslem women living in an urban area of District Lucknow, India were analyzed to determine duration and frequency of breast feeding, child spacing, and their beliefs about advantages and disadvantages of breast feeding. Physicians attended 78.9% of the deliveries. Nurse-midwives and trained traditional birth attendants delivered the remaining infants. The woman had a average 5.9 children. All women breast fed their infants from day 1. The mothers were more likely to breast feed sons longer than daughters (24.9 months vs. 21.3 months). Median breast feeding duration was 22.4 months compared with 23.8 months for Moslem women in Bangladesh. It declined with parity except for the 1st child which was 23 months and the 5th child which was 17 months (26.7 months for parity 2, 25.6 for parity 3, 14.8 for parity 4, and 11 for parity 6). Infants 3 months old received more feeds each day than those 3 months old (7.5 vs. 5.3). 50% of mothers who breast fed for 12-17 months had a 2-year birth interval and those who breast fed for 2 years had an interval of about 4 years. While 100% of the noneducated mothers and 72.7% of the educated mothers knew that breast feeding maintains a child's health only 36.3% of educated mothers and none of the noneducated mothers knew that it conferred immunity. Mothers began giving their infants other liquids at a mean of 1.4 months. These liquids were water, diluted milk, toned milk, barley water, and pulse's water. The average age for introduction of solid foods (rice, kheer, porridge, bread, biscuits, boiled eggs, egg yolks, bananas, pudding, curd, and wheat) was 8.2 months.^ieng