Early Stress History Alters Serum Insulin-Like Growth Factor-1 and Impairs Muscle Mitochondrial Function in Adult Male Rats.

Early-life adversity is associated with an enhanced risk for adult psychopathology. Psychiatric disorders such as depression exhibit comorbidity for metabolic dysfunction, including obesity and diabetes. However, it is poorly understood whether, besides altering anxiety and depression-like behaviour, early stress also evokes dysregulation of metabolic pathways and enhances vulnerability for metabolic disorders. We used the rodent model of the early stress of maternal separation (ES) to examine the effects of early stress on serum metabolites, insulin-like growth factor (IGF)-1 signalling, and muscle mitochondrial content. Adult ES animals exhibited dyslipidaemia, decreased serum IGF1 levels, increased expression of liver IGF binding proteins, and a decline in the expression of specific metabolic genes in the liver and muscle, including Pck1, Lpl, Pdk4 and Hmox1. These changes occurred in the absence of alterations in body weight, food intake, glucose tolerance, insulin tolerance or insulin levels. ES animals also exhibited a decline in markers of muscle mitochondrial content, such as mitochondrial DNA levels and expression of TFAM (transcription factor A, mitochondrial). Furthermore, the expression of several genes involved in mitochondrial function, such as Ppargc1a, Nrf1, Tfam, Cat, Sesn3 and Ucp3, was reduced in skeletal muscle. Adult-onset chronic unpredictable stress resulted in overlapping and distinct consequences from ES, including increased circulating triglyceride levels, and a decline in the expression of specific metabolic genes in the liver and muscle, with no change in the expression of genes involved in muscle mitochondrial function. Taken together, our results indicate that a history of early adversity can evoke persistent changes in circulating IGF-1 and muscle mitochondrial function and content, which could serve to enhance predisposition for metabolic dysfunction in adulthood.

Molecular-level removal of proteinaceous contamination from model surfaces and biomedical device materials by air plasma treatment.

Established methods for cleaning and sterilising biomedical devices may achieve removal of bioburden only at the macroscopic level while leaving behind molecular levels of contamination (mainly proteinaceous). This is of particular concern if the residue might contain prions. We investigated at the molecular level the removal of model and real-life proteinaceous contamination from model and practical surfaces by air plasma (ionised air) treatment. The surface-sensitive technique of X-ray photoelectron spectroscopy (XPS) was used to assess the removal of proteinaceous contamination, with the nitrogen (N1s) photoelectron signal as its marker. Model proteinaceous contamination (bovine serum albumin) adsorbed on to a model surface (silicon wafer) and the residual proteinaceous contamination resulting from incubating surgical stainless steel (a practical biomaterial) in whole human blood exhibited strong N1s signals [16.8 and 18.5 atomic percent (at.%), respectively] after thorough washing. After 5min air plasma treatment, XPS detected no nitrogen on the sample surfaces, indicating complete removal of proteinaceous contamination, down to the estimated XPS detection limit 10ng/cm(2). Applying the same plasma treatment, the 7.7at.% nitrogen observed on a clinically cleaned dental bur was reduced to a level reflective of new, as-received burs. Contact angle measurements and atomic force microscopy also indicated complete molecular-level removal of the proteinaceous contamination upon air plasma treatment. This study demonstrates the effectiveness of air plasma treatment for removing proteinaceous contamination from both model and practical surfaces and offers a method for ensuring that no molecular residual contamination such as prions is transferred upon re-use of surgical and dental instruments.

Study of age of fusion of hyoid bone.

The present study was conducted to determine the age of fusion of greater cornua with the body of hyoid bone. Age of fusion of hyoid bone can help in determining the age of an individual, especially of unknown dead bodies where only skeletal remains are available. A victim of compression of neck will more likely have fracture of hyoid bone if his hyoid bone is fused. Indian authors have reported that the fusion of hyoid bone occurs after 40 years of age. Studies done by foreign workers observed that hyoid bone fused at an earlier age (30-40 years). A total of 170 excised hyoid bones from dead bodies belonging to the age group of 20-65 years were studied. Fusion occurred earlier in females as compared to males by about 5 years. The mean age of unilateral and bilateral fusion in males was 38.25 and 53.16 years, respectively. The mean age of unilateral and bilateral fusion in females was 38.00 and 48.50 years, respectively. All the hyoid bones were fused after the age of 60 years. No significant differences were found between the fusion on right and left side.

Pattern of injuries in fatal falls from buildings.

When bodies are brought for autopsy it is sometimes unclear whether the injuries are due to a fall from a height or due to blunt trauma from other causes, especially when the bodies are found near buildings with no eyewitnesses available. Studies discussing the injury patterns in adults after falls exclusively from buildings are rare. A five-year retrospective study was carried out on 151 cases of fatal falls from buildings brought for autopsy. The aim was to assess the pattern of these injuries and identify features helpful in discriminating between these and injuries due to blunt trauma from other causes. The majority of cases comprised subjects who fell from heights of 10-20 feet (3-6m) with most falls occurring late at night or in the early morning. The pattern that emerged is quite distinct from ground level falls and pedestrian injuries. Abrasions are the commonest injury and bruises very rare. Lacerations are mostly on the head and skull fractures are evenly distributed between the vertex, base and vertex plus base. Subarachnoid haemorrhage is the commonest intracranial lesion. Extradural haemorrhage alone is rare. Fractures of ribs and cervical vertebrae are common and fractures of thoracic vertebrae and long bones are uncommon. Safety measures to prevent such falls have been suggested.

Risk assessment of cardiovascular diseases among bank employees--a biochemical approach.

In different epidemiological studies, an association between sedentary life style and incidence of cardiovascular diseases has been demonstrated. This study was carried out among 50 sedentary bank employees working at different nationalized banks within Calcutta City, India and 50 individuals with non-sedentary jobs matched for age group and sex for the control group. Body mass index (BMI), blood pressure (BP), plasma level of glucose, total cholesterol, low density lipoprotein (LDL), very low density lipoprotein (VLDL) and triglycerides (Tgs) were found to be significantly higher whereas high density lipoprotein (HDL) and reduced glutathione were found to be significantly lower among the bank employees as compared with the control subjects. Further, significant alteration in the activity of different enzymes (e.g., glutathione reductase, glutathione S-transferase, glucose 6 phosphate dehydrogenase, catalase and superoxide dismutase) involved in glutathione-mediated antioxidant scavenging systems and increases in lipid peroxidation were also observed in the bank employees. From the study, it appears that the relative risk for cardiovascular disease is increased among bank employees due to the sedentary nature of their jobs.

Study of head injury victims in fatal road traffic accidents in Delhi.

The study revealed 31% were the victims of Head injury injury in this part of Delhi in vehicular accidents. Although majority of the fatalities were on the spot quite a number of such victims survived for a varied period. A multipronged approach including the preventive and curative measures for this man-made calamity is the need of the hour. Amongst various preventive measure are wearing helmet be made compulsory for both the driver and pillion riders of two-wheelers, development of a safe traffic sense amongst the road users. Traffic Police should be honest, exemplary punishment for the reckless driver, decongestion of Delhi roads etc. The curative measures are facilities of CT scanners and neurosurgeons in all the major hospitals, provision of first-aid in the PCR (Police control room) vans, to institute immediate treatment to the victim without waiting for the medico-legal formalities, opening of more fully equipped Accident and trauma centres. It is high time the concerned authorities gave some serious thought towards this avoidable epidemic on Delhi roads.

Influence of cigarette smoking on Vitamin C, glutathione and lipid peroxidation status.

There has been a growing interest during recent years in the role of free radicals and lipid-peroxidation at tissue-level for the causation of cancer and other age-related diseases like atherosclerosis, rheumatoid arthritis, cataract etc. Free radicals and increased lipid peroxidation play a significant role for causation of human diseases by oxidative damage and functional degeneration of the tissues. Vitamin C, a well-known dietary antioxidant, and other enzymatic antioxidants like glutathione can protect the lipids of lipoproteins and other biomembranes against peroxidative damage by intercepting oxidants before they can attack the tissues. But cigarette smoking was found to affect the antioxidant protective action of Vitamin C, glutathione etc. A group of adult male smokers in this study were found to have lowered Vitamin 'C' & glutathione levels, but increased lipid-peroxide levels in their blood. Thus the increased pathogenicity of the smoking may also be due to indirect biochemical effect of enhanced oxidative stress by increased lipid-peroxidation and lowered Vitamin C & other antioxidants at tissue-level.

Estimation of age from epiphyseal union at the wrist and ankle joints in the capital city of India.

Various studies conducted in and outside India for estimation of age from long bone joints revealed that unlike other vital parameters no uniform pattern exists for epiphyseal union of long bones in different countries of the world and also in different parts of the same country. A radiological study was conducted in 180 boys and girls of the capital city of India, Delhi, between the age group of 14-20 years to find out the age at which epiphyseal union at wrist and ankle joint takes place. The result of the study showed that the wrist joint epiphyseal union is completed in all cases in the age group of 19-20 years in males and 18-19 years in females. In the ankle joint, 17-18 years was the age group in males and 16-17 years the age group in females which showed complete epiphyseal union in all cases.

Effect of thiocyanate ingestion through milk on thyroid hormone homeostasis in women.

A thyroid-hormonal evaluation of thirty-five women consuming commercially packed milk containing thiocyanate was carried out. The mean serum thiocyanate concentration, which was measured by the FeCl3 colour test, was significantly higher (P < 0.01) than that of control subjects. Serum thyroxine (T4), triiodothyronine (T3) and thyroid-stimulating hormone (TSH) concentrations of exposed women were compared with those of thirty-five control subjects. Thiocyanate ingestion was associated with lower levels of T4 (P < 0.01) and higher levels of TSH (P < 0.01) compared with the control subjects. T3 was found to be higher in the women consuming thiocyanate-containing milk but the difference was not significant. The serum T4 level was found to be negatively correlated (r -0.359, P < 0.05) while the TSH level was positively correlated (r 0.354, P < 0.05) with thiocyanate concentration in the exposed group. From this study, it appears that ingestion of milk with added thiocyanate impairs thyroid function.

Evaluation of cyanide exposure and its effect on thyroid function of workers in a cable industry.

A thyroid-hormone evaluation of workers dealing with cyanide compounds in an electroplating process of a cable industry was carried out. Serum thiocyanate (SCN) levels of 35 nonsmoking copper-ply employees were assayed by a ferric-chloride color test. The mean SCN concentration of these employees was 316 +/- 15 mumol/L, which was significantly (P < 0.01) higher than that of control subjects (90.8 +/- 9.02 mumol/L). Serum thyroxine (T4), triiodothyronine (T3), and thyroid-stimulating hormone (TSH) concentrations of exposed workers were compared with those of 35 control subjects. Cyanide exposure resulted in a decrease in T4 and T3 concentrations (P < 0.05) and an increase in TSH concentration (P < 0.05), compared with the control subjects. The serum T4 level was found to be negatively correlated (r = -0.363, P < 0.05), whereas the TSH level was positively correlated (r = 0.354, P < 0.05), with SCN concentration in the exposed group. The study suggests that occupational cyanide exposure in the industry impairs thyroid function.

Carbohydrate-mediated regulation of interaction of Vibrio cholerae hemolysin with erythrocyte and phospholipid vesicle.

Vibrio cholerae hemolysin is an extracellular pore-forming monomeric protein with a native molecular weight of about 60,000. In this study, we showed that the hemolysin interacted with immobilized phospholipids and cholesterol and formed oligomers in vesicles constituted from phospholipids alone with a stoichiometry identical to those produced in rabbit erythrocyte membrane. However, the hemolysin bound to glycoproteins with terminal beta1-galactosyl residues and an association constant of 9.4 x 10(7) M(-1) was estimated for the hemolysin-asialofetuin complex by solid phase binding assay. Oligomerization of the hemolysin in lipid bilayer converted the sugar-binding monomer to a lectin with strong carbohydrate-dependent hemagglutinating activity accompanied by inactivation of hemolytic activity and loss in ability to interact with phospholipids. There was no evidence for erythrocyte surface carbohydrates playing an essential role in interaction of the hemolysin with the cell. However, specific glycoproteins inhibited hemolysis of rabbit erythrocytes as well as interaction of the hemolysin with phospholipid. The results suggest (i) V. cholerae hemolysin is a monomer with distinct domains associated with specific binding to carbohydrates and interaction with lipids, (ii) the pore-forming property depends solely on the protein-lipid interaction with no evidence for involvement of sugars, and (iii) specific sugars can down-regulate the ability of the hemolysin to form pores in lipid bilayers.

Effects of human placental extract on brain monoamines and monoamine oxidase activity in rats.

Human placental extract, an agent clinically used world-wide in a number of physiological anomalies, has been claimed to be effective in children of slow learners. Since the monoaminergic neurotransmitter systems in the brain play an important role in the processes of learning and memory, we examined the effects of human placental extract on the levels of norepinephrine, dopamine and serotonine in rat brain as an attempt to evaluate the possible underlying biochemical mechanism of action of the extract. We also determined the changes of brain monoamine oxidase (MAO) activity following placental extract treatment. The results showed that subchronic (5, 10, 15 or 20) administration of placental extract (2-4 ml/kg/day) had the effect of increasing all the monoamines and decreasing the MAO activity which could be the possible mode of action of the extract in slow learners.

L-rhamnose inhibits proliferation of murine splenocytes by the lipopolysaccharide and polysaccharide moiety of Shigella dysenteriae type 1 lipopolysaccharide.

The induction of proliferation of murine splenocytes by lipopolysaccharide (LPS) of Shigella dysenteriae type 1 and its polysaccharide (PS) and lipid A fractions was investigated. The LPS-induced proliferation reached a maximum at a concentration of 30 ng/ml. The PS and lipid A induced proliferation of murine splenocytes at similar concentrations. Preincubation of murine splenocytes with varying concentrations of L-rhamnose blocked LPS- and PS-induced proliferation in a dose-dependent manner. The lipid A-induced stimulation, on the contrary, was not affected by preincubation of the cells with L-rhamnose. These data suggest that activation of splenocytes by LPS and PS is mechanistically different from that induced by lipid A and is presumably involved in the specific recognition of carbohydrate structures on LPS and PS.

Carbohydrate-dependent binding of the cell-free hemagglutinin of Vibrio cholerae to glycoprotein and glycolipid.

The carbohydrate-binding specificity of the cell-free hemagglutinin (HA) of Vibrio cholerae (K.K. Banerjee, A.N. Ghose, K. Datta-Roy, S.C. Pal, and A.C. Ghose, Infect. Immun.58:3698-3705, 1990) was studied by using glycoconjugates with defined sugar sequences. The HA was not inhibited by simple sugars including glucobiose, galabiose, and their N-acetylated derivatives. The hemagglutination of rabbit erythrocytes by the HA was inhibited moderately by fetuin, calf thyroglobulin, and human alpha 1-acid glycoprotein, all of which contain multiple asparagine-linked complex-type oligosaccharide units alone or in combination with serine/threonine-linked oligosaccharide units. The inhibitory potencies of the glycoproteins increased approximately 10-fold following removal of the terminal sialic acid and were completely destroyed by exhausative proteolysis. The HA agglutinated phosphatidylcholine liposomes containing GM1-ganglioside or its asialo-derivative in the presence of Ca2+ ions. The association constants of the complexes of the HA with asialofetuin, asialothyroglobulin, GM1-ganglioside, and asialo-GM1-ganglioside were determined by an enzyme-linked immunosorbent assay-based assay and found to be 1.7 x 10(7) M-1, 1.5 x 10(7) M-1, 1.8 x 10(7) M-1, and 2.4 x 10(7) M-1, respectively. Studies using chemically modified glycoproteins and plant lectins with defined sugar specificity revealed that the HA recognized the terminal beta 1-galactosyl moiety of these glycoconjugates. There was no evidence for the presence of an extended carbohydrate-binding domain in the HA molecule or a preference of the HA for a complex, branched oligosaccharide structure. Similar to the mechanisms proposed for the binding of cholera toxin and Shiga toxin to glycolipids and neoglycoproteins, the strong interaction of V. cholerae cell-free HA with glycoconjugates appeared to be a consequence of multiple weak binding to terminal beta1-galactosyl moieties of the glycoproteins or glycolipids.

Effects of human placental extract on hepatic drug metabolizing enzyme.

Effects of acute or subchronic administration of human placental extract (HPE), a worldwide clinically used agent, on hepatic drug metabolizing enzyme activities were evaluated in rats. Hepatic microsomal cytochrome P-450 (Cyt. P450) and cytochrome b5 (Cyt. b5) contents and cytosolic glutathione S-transferase (GST) activities were maximally induced after various periods of time following a single intraperitoneal injection of HPE (4 ml/kg) whereas microsomal UDP-glucuronyltransferase (UDPGT) activities were inhibited significantly. All these altered effects were returned almost to the basal levels after 96 h of treatment. Subchronic treatment (30 days) with HPE (1,2 or 4 ml/kg) afforded a significant induction of Cyt. P-450 and Cyt. b5 levels and that of GST activities with a concurrent suppression of the activities of UDPGT and these results were found to be dose-dependent. However, microsomal NADPH cytochrome c reductase activity was not affected either by acute or subchronic treatment. The observed variations in the levels and activities of above house-keeping enzymes were discussed in relation to the possible carcinogenic risk of long-term treatment with this pharmaceutical agent.

Role of human placental extract on succinic dehydrogenase activity in carrageenin-induced edema in rats in vivo and its effect on erythrocyte lysis, platelet aggregation and trypsin activity in vitro.

Significant increase of liver succinic dehydrogenase (SDH, EC 1.3.99.1) activity was produced by carrageenin-induced edema in rats. Pretreatment with human placental extract inhibited the increased liver SDH activity in a dose-dependent manner. Placental extract was found to have little or no effect on the liver SDH activity in normal rats. Furthermore, heat-induced erythrocyte lysis was inhibited to a substantial extent by the extract and was found to be dose-responsive. However, adenosine diphosphate (ADP)-induced platelet aggregation and trypsin activity were not changed by the placental extract in vitro. The study indicates that the membrane stabilization and depletion of adenosine triphosphate (ATP) synthesis may contribute to antiinflammatory effect of the extract.

Effect of cigarette smoking on thyroid hormone homeostasis.

Serum thyroxine (T4), triiodothyronine (T3), thyroid stimulating hormone (TSH) and thiocyanate (SCN) levels in 40 smokers smoking filter or non-filter cigarettes were compared to those in 20 control subjects. Both types of cigarette smoking resulted in a decrease in T4 (P < 0.01) and increase in TSH (P < 0.05) to the same degree as compared to control. T3 was found to be higher in the smokers but the rise was not significant. SCN was elevated significantly in the smokers. The filter attached to the cigarette was found to have little or no effect on the alteration of the levels of the biochemical indices studied.

Elevated lipid peroxidation, decreased glutathione levels and changes in glutathione-related enzymes in rats treated with human placental extract.

The in vivo effects of human placental extract (1-4 ml/kg) on hepatic lipid peroxidation, blood and liver glutathione (GSH) levels and several enzymes associated with the antioxidant defence mechanism; i.e., catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase, together with some blood biochemical responses were investigated in rats. At an optimal dose level (4 ml/kg), a single acute intraperitoneal administration of the extract caused a significant enhancement (49.9%; p < 0.001) of lipid peroxidation with a decline in GSH level both in blood (45.1%; p < 0.001) and liver (61.0%; p < 0.001) in comparison to control animals. Activities of catalase, glutathione peroxidase and glutathione reductase were inhibited in a dose-responsive way by the treatment with the extract which also increased the activity of glutathione S-transferase in a dose-dependent manner. The extract was found to be hepatotoxic in terms of elevation of serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, serum lactate dehydrogenase and blood methemoglobin concentration. Results of this study suggest the adverse consequences of the administration of the extract due to its substantial ability to alter normal cellular processes.

Anti-inflammatory effect of human placental extract: a biochemical mechanistic approach.

Significant increase of liver succinic dehydrogenase (SHD) activity was produced by carrageenin-induced edema in rats. Pretreatment with human placental extract "Placentrex" inhibited the increased liver SHD activity in a dose-dependent manner. "Placentrex" was found to have no effect on the liver SHD activity in normal rats. Furthermore, heat-induced erythrocyte lysis was inhibited to a substantial extent by "Placentrex" and was found to be almost dose-responsive. However, adenosine diphosphate (ADP)-induced platelet aggregation and trypsin activity were not changed in vitro by the "Placentrex". No alkaline phosphatase activity was found in this preparation. All these studies indicate that the membrane stabilization and depletion of adenosine triphosphate (ATP) synthesis may be the basis of anti-inflammatory effect of this drug.

Purification and characterization of a novel hemagglutinin from Vibrio cholerae.

A lectin with strong hemagglutinating activity toward erythrocytes of several animal species was isolated from an 18-h culture supernatant of a diarrheagenic strain, V2, of non-O1 Vibrio cholerae. The hemagglutinin (HA) was purified free of lipopolysaccharide by salt fractionation followed by gel filtration, hydrophobic interaction chromatography, and, finally, gel filtration in the presence of urea and deoxycholate. The purification procedure resulted in an HA preparation with 80-fold enhancement of specific activity. The HA consisted of noncovalently bound subunits of Mr 62,000 and behaved essentially as a single component with pI 6.0. Nonpolar and acidic amino acids contributed 46 and 24%, respectively, to the total amino acid residues. Electron micrographs of the HA showed it to consist of large, nonstoichiometric aggregates' of disklike molecules of 10-nm diameter. Inhibition of the HA by the glycoproteins fetuin, asialofetuin, and mucin, but not by ovalbumin and simple sugars, suggested the specific requirement of complex carbohydrates for binding. Rabbit antisera to the purified HA inhibited the hemagglutinating activities of the crude cell-free HA preparations, but not cell-associated HA activities of the parent (V2) or of other O1 and non-O1 V. cholerae strains. This suggested that the released and cell-associated HA activities were mediated by antigenically distinct components. Immunoblotting experiments showed that the antisera recognized a polypeptide component of Mr 62,000 in the cell envelope preparations of the parent and several other V. cholerae O1 and non-O1 strains. These data suggested that the HA was a nonfimbrial lectin of somatic origin with no protease activity and was apparently distinct from V. cholerae HAs described so far.