Comparative sperm protein profiling in bulls differing in fertility and identification of phosphatidylethanolamine-binding protein 4, a potential fertility marker.

This study aimed to identify sperm proteomic signatures regulating sperm functions and fertility by: (i) comparing the sperm electrophoretic protein profiles and identifying the differentially abundant proteins among breeding bulls differing in fertility status and (ii) elucidating the possible role of one of the identified novel proteins, PEBP4 on sperm function and fertility. The grouping of bulls as fertile (n = 6) and low fertile (n = 6) was performed based on bull fertility index and infertile (n = 6) based on semen rejection rate (>33%). The sperm motility, fructolysis index, acrosomal reaction, intracellular calcium levels, and seminal plasma fructose and calcium levels were studied among fertility groups. The differentially expressed sperm proteins observed in single- and two-dimensional gel electrophoresis (2DE) were identified using Nano-LC-MS/MS. In the fertile bulls, the expression levels of calmodulin (CALM1), spermadhesinZ13 (SPADH2), and phosphatidylethanolamine-binding protein 4 (PEBP4) were significantly (p < 0.05) higher than in other fertility groups. In bovine, expression of PEBP4 a novel seminal protein was not observed in spermatozoa of infertile bulls. When the bulls were grouped based on the presence (n = 8) or absence (n = 10) of PEBP4 protein in spermatozoa, a positive significant (p < 0.05) association of this protein with the percentage of motile, type-A spermatozoa, and sperm fructose uptake was observed. Further, PEBP4 was localized in elongated spermatids, Leydig cells, excurrent duct system, and principal piece of spermatozoa. These findings suggest a crucial role for the PEBP4 protein in spermiogenesis, epididymal sperm maturation, and sperm motility. This first study in bovine indicates the positive association of PEBP4 in regulating sperm maturation, functions, and fertility and could be a potential marker for predicting semen quality and fertility.

Determination of ground and excited state dipole moments of dipolar laser dyes by solvatochromic shift method.

The absorption and fluorescence spectra of three medium sized dipolar laser dyes: coumarin 478 (C478), coumarin 519 (C519) and coumarin 523 (C523) have been recorded and studied comprehensively in various solvents at room temperature. The absorption and fluorescence spectra of C478, C519 and C523 show a bathochromic and hypsochromic shifts with increasing solvent polarity indicate that the transitions involved are π→π(∗) and n→π(∗). Onsager radii determined from ab initio calculations were used in the determination of dipole moments. The ground and excited state dipole moments were evaluated by using solvatochromic correlations. It is observed that the dipole moment values of excited states (µe) are higher than corresponding ground state values (µg) for the solvents studied. The ground and excited state dipole moments of these probes computed from ab initio calculations and those determined experimentally are compared and the results are discussed.

Changes in the activity levels of glutamine synthetase, glutaminase and glycogen synthetase in rats subjected to hypoxic stress.

Exposure to high altitude causes loss of body mass and alterations in metabolic processes, especially carbohydrate and protein metabolism. The present study was conducted to elucidate the role of glutamine synthetase, glutaminase and glycogen synthetase under conditions of chronic intermittent hypoxia. Four groups, each consisting of 12 male albino rats (Wistar strain), were exposed to a simulated altitude of 7620 m in a hypobaric chamber for 6 h per day for 1, 7, 14 and 21 days, respectively. Blood haemoglobin, blood glucose, protein levels in the liver, muscle and plasma, glycogen content, and glutaminase, glutamine synthetase and glycogen synthetase activities in liver and muscle were determined in all groups of exposed and in a group of unexposed animals. Food intake and changes in body mass were also monitored. There was a significant reduction in body mass (28-30%) in hypoxia-exposed groups as compared to controls, with a corresponding decrease in food intake. There was rise in blood haemoglobin and plasma protein in response to acclimatization. Over a three-fold increase in liver glycogen content was observed following 1 day of hypoxic exposure (4.76 +/- 0.78 mg.g-1 wet tissue in normal unexposed rats; 15.82 +/- 2.30 mg.g-1 wet tissue in rats exposed to hypoxia for 1 day). This returned to normal in later stages of exposure. However, there was no change in glycogen synthetase activity except for a decrease in the 21-days hypoxia-exposed group. There was a slight increase in muscle glycogen content in the 1-day exposed group which declined significantly by 56.5, 50.6 and 42% following 7, 14, and 21 days of exposure, respectively. Muscle glycogen synthetase activity was also decreased following 21 days of exposure. There was an increase in glutaminase activity in the liver and muscle in the 7-, 14- and 21-day exposed groups. Glutamine synthetase activity was higher in the liver in 7- and 14-day exposed groups; this returned to normal following 21 days of exposure. Glutamine synthetase activity in muscle was significantly higher in the 14-day exposed group (4.32 mumol gamma-glutamyl hydroxamate formed.g protein-1.min-1) in comparison to normal (1.53 mumol gamma-glutamyl hydroxamate formed.g protein-1.min-1); this parameter had decreased by 40% following 21 days of exposure. These results suggest that since no dramatic changes in the levels of protein were observed in the muscle and liver, there is an alteration in glutaminase and glutamine synthetase activity in order to maintain nitrogen metabolism in the initial phase of hypoxic exposure.

Nutritional status of children of urban leprosy patients staying at preventoria based on biochemical parameters.

Measurements of nutritionally relevant biochemical and endocrine variables were made on 60 apparently healthy children (group A) whose parents suffered from leprosy and who had been separated at the age of 4 years and brought up in preventoria. Most of the measurements were also made on a comparison group of healthy children from the same poor socio-economic class (group B). In both groups the serum concentrations of cholesterol and triglycerides were well below those found in Western populations. Almost all the children in both groups were anaemic, but serum iron and ferritin levels were satisfactory. Folate and vitamin B12 levels were measured in group A only and were low in a significant proportion. Deficiency of these water-soluble vitamins may be a cause of the anaemia. Low albumin levels were found in 40% of group A children, compared with 2% in group B. The concentrations of calcium and magnesium were lower and that of phosphate higher in group A than in B. In both groups one-third of the children had low levels of serum zinc. Fifteen per cent of group A children had biochemical evidence of vitamin A deficiency, but none were deficient in vitamin E. Levels of total T3 and total T4 were below the lower limit of normal in a substantial proportion of children in both groups. Concentrations of parathyroid hormone were increased in parallel with the low values for serum calcium. Radiological studies of ossification centres in 57 group A children showed delayed maturation in 11 cases. The relevance of these findings to previous studies of the children of lepers in India is discussed.

Role of xanthinol nicotinate in the revival of monkeys subjected to acute haemorrhagic shock.

The therapeutic potential of xanthinol nicotinate in the revival of anaesthetised monkeys subjected to acute blood loss was investigated. The arterial pressure was lowered to 40 +/- 5 mmHg by rapid arterial bleeding and was maintained at this level for 2 h. Shed blood was then returned through infusion, to the animals. Animals alive at the end of 72 h observation period were considered as survivors. The test drug was infused 1/2 h prior to and 1/2 h, 1 h, 1 1/2 h and 2 h after the onset of oligaemic hypotension. The animals which received normal saline instead of test drug were treated as control. The physiological and biochemical parameters recorded prior to and after the onset of oligaemic hypotension were heart rate, pulse pressure, electrocardiogram, electroencephalogram (EEG), lactic acid, creatine phosphokinase, urea and glucose. The results showed tachycardia, narrowing of pulse pressure, depression of ST segment with occasional T inversion, slowing of EEG with increase in amplitude, rise in blood lactic acid, creatine phosphokinase, urea and glucose. The magnitude of these responses were proportional to the duration and severity of shock. These changes were markedly attenuated in the drug treated group. Enhancement of survival was observed in drug treated groups as compared to control. It was 10 per cent in control as against 60, 86, 71, 57 and 50 per cent in the groups which received the test drug 1/2 h prior to and 1/2 h, 1 h, 1 1/2 h and 2 h after the onset of oligaemic hypotension. It is concluded that the beneficial effect of the drug in the revival of monkeys subjected to acute haemorrhagic shock may be due to better maintenance of tissue perfusion.

Effects of pyridoxine supplementation on blood glucocorticoids level, aspartate aminotransferase activity and glucose tolerance pattern under acute hypoxic stress.

Studies were conducted on male adult rabbits to find out the changes in blood glucocorticoid levels along with the changes in aspartate aminotransferase activity in blood and the role of pyridoxine on the glucose tolerance pattern under hypoxic stress. Hypoxic stress was produced by exposing the animals to a simulated altitude of 7,000 m for 6 h. In the first set of experiments 10 rabbits were used. Blood haemoglobin level, plasma and erythrocyte glucocorticoid levels and erythrocyte GOT activity were measured just before and after the exposure to hypoxia. Erythrocyte GOT activity was measured both without and with 50 mg of pyridoxal phosphate addition to the incubation mixture. Glucocorticoid levels in plasma increased by 11% whereas in erythrocytes the increase was 55% after hypoxia. Percent stimulation of erythrocyte GOT activity with pyridoxal phosphate before exposure to hypoxia was 180% but increased to 321% after exposure. In the second set of experiments another 10 rabbits were used. First they were exposed to hypoxia without pyridoxine hydrochloride feeding and then after 7 days with 3 mg of pyridoxine hydrochloride feeding. For glucose tolerance tests the animals were fed with 1 g of glucose immediately after the hypoxic exposures. Plasma reduced glutathione (GSH), LDH and ICDH activities increased and GOT activity was depressed after hypoxic stress, but when the animals were fed pyridoxine hydrochloride prior to the exposure the enzyme activities remained unaltered after hypoxic stress. Pyridoxine hydrochloride did not alter the pattern of glucose tolerance after hypoxic stress.

Biochemical effects of chronic exposure to noise in man.

Biochemical parameters in 75 normal healthy male subjects exposed to intense noise of 88-107 dB(A)(6-8 h/day) for 10 to 15 years during their work situation have been monitored and compared with 35 normal unexposed subjects. Levels of free cholesterol (P less than 0.001), gamma-globulin (P less than 0.01) and cortisol (P less than 0.01) were found to be significantly higher in the exposed subjects. Significant changes in free cholesterol also altered the ratio of free to esterified cholesterol significantly (P less than 0.001). The value of the A/G ratio was also lower in the exposed group. Uric acid did not show any change. The study shows that in the exposed group the esterification process of cholesterol was modified. There was an effect on pituitary-adrenal axis as well as host-defence mechanism.

Determination of neptunium by redox titration.

A simple and quick method for the potentiometric determination of neptunium on the 2-5 mg scale has been developed. It consists of oxidation to Np(VI) by AgO or fuming with HClO(4), destruction of excess of AgO by sulphamic acid, reduction of Np(VI) to Np(IV) with a slight excess of standard Fe(II) in 2M H(2)SO(4) and potentiometric titration of the excess of Fe(II) with standard Ce(IV). The precision is +/-0.5%.