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Unsatisfactory performance of the existing BCG vaccines, especially against the adult pulmonary disease, has urged the need for an effective vaccine against tuberculosis (TB). In this study, we employed differential proteomics to obtain a list of antigens as potential vaccine candidates. Bacterial epitopes being presented at early stages on MHC class I and class II molecules of macrophages infected with Mycobacterium tuberculosis (M. tb) were identified using iTRAQ labelling and reverse phase LC-MS/MS. The putative vaccine candidates, thus identified, were tested as plasmid DNA vaccines in mice to ascertain their protective efficacy against the aerosolized M. tb challenge, based on their ability to reduce the bacterial load in the lungs of infected mice. Here, we observed that 4 out of the 17 selected antigens imparted significant protection against the challenge of M. tb. The four shortlisted antigens were further assessed in a more stringent guinea pig model, where too, they demonstrated.significant protection. It concludes that combining a proteomics approach with the in vivo assessment of vaccine candidates in animal models can be valuable in identifying new potential candidates to expand the antigenic repertoire for novel vaccines against TB.

Sorghum is one of the commercially feasible lignocellulosic biomass and has a great potential of being sustainable feedstock for renewable energy. As with any lignocellulosic biomass, sorghum also requires pretreatment which increases its susceptibility to hydrolysis by enzymes for generating sugars which can be further fermented to alcohol. In the present study, sorghum biomass was evaluated for deriving maximum fermentable sugars by optimizing various pretreatment parameters using statistical optimization methods. Pretreatment studies were done with H2SO4, followed by enzymatic saccharification. The efficiency of the process was evaluated on the basis of production of the total reducing sugars released during the process. Compositional analysis was done for native as well as pretreated biomass and compared. The biomass pretreated with the optimized conditions could yield 0.408 g of reducing sugars /g of pretreated biomass upon enzymatic hydrolysis. The cellulose content in the solid portion obtained after pretreatment using optimised conditions was found to be increased by 43.37% with lesser production of inhibitors in acid pretreated liquor.

The anti proliferative potential of siRNA26, targeted to Aurora kinase B, in prostate cancer cells is known from a previous study from our laboratory. Here we first show that siRNA26 cleaves at the same position of the target mRNA in the prostate cancer and hepatocellular carcinoma cell lines, PC3 and HepG2 respectively. Aurorakinase B specific siRNA, but not a control siRNA, inhibited PC3 and HepG2 cell proliferation and cell migration. These effects correlated to RNA silencing of Aurorakinase B in both the cell lines. Intra-tumoral administration of HiPerfect complexed siRNA26 inhibited the growth of HepG2 xenografts in SCID mice. In an orthotopic setting, intravenous administration of HiPerfect encapsulated siRNA26 appeared to reduce the severity of multifocal lesions.

Administration of cadmium (Cd) significantly increased the peroxidation markers such as malondialdehyde and protein carbonyls along with significant decrease in antioxidant markers such as super oxide dismutase and reduced glutathione in liver and kidney tissues. Cadmium also caused a significant alteration in hepatic and renal functional markers in serum viz. total protein, albumin, alanine transaminase, blood urea nitrogen and creatinine. Prominent pathological changes observed in liver were severe vascular and sinusoidal congestion with diffuse degenerative changes and mononuclear infiltration into peripheral areas, while the kidney showed vascular and glomerular congestion, cloudy swelling of tubular epithelium. Coadministration of ethonolic extract of T. terrestris or vitamin E along with Cd significantly reversed the Cd induced changes along with significant reduction in Cd load.

Significant increase in the salivary gland weight was observed after exposure to single therapeutic dose of 3.7 MBq of 131I in mice. Pre-supplementation of antioxidants, O. sanctum leaf extract, turmeric extract and vitamin E for 15 days before 131I exposure demonstrated significant reduction in the salivary gland weight. No major histopathological changes were observed in the salivary gland of experimental animals at 24 h of exposure. Micronuclei index in the bone marrow of polychromatic (PCEs) and normochromatic erythrocytes (NCEs) remained unchanged in all the experimental groups. However, PCE/NCE ratio in the bone marrow decreased significantly in all the 131I exposed animals irrespective of antioxidant supplementation status. The normalization of salivary gland weight by antioxidant pre-supplementation in radioiodine exposed mice is suggestive of the possible ameliorating effect of antioxidants on the salivary gland weight recommending further detailed studies regarding the functional aspect of the salivary gland in higher animals.

With a view to examine the effect of chronic maternal stress on cognitive function in the offspring during young age, pregnant Wistar rats were subjected to restraint stress from embryonic day 11 till delivery. Male and female pups born to these stressed rats were subjected to passive avoidance test on postnatal day 30 and 31. Results were compared with rats of the same age and sex born to control mothers, which were not stressed. The results showed that prenatal maternal restraint stress impairs the memory retention during young age in both sexes. The memory retention deficit induced by maternal restraint stress was evident in the decreased latency to enter the dark compartment of passive avoidance apparatus by the rats born to stressed mothers. The observed behavioral deficit may be due to the insult of stress on the developing hippocampus, a structure of the brain concerned with learning and memory. The results suggest that prolonged prenatal stress leads to long lasting malfunction in the behavioral development during young age in both male and female young rats. However when compared to their respective stress naïve controls, it seems evident that prenatal restraint stress has a less effect on females which could be due to their oesterogenic effects. These data reinforce the view that prenatal stress affects cognitive development in a sex-specific manner.

Epilepsy is a common health problem. Although variety of factors influence the incidence and prevalence of seizures, cytokines are considered to play an important role in seizures. Cytokines are also known to be involved in other neurodegenerative disorders. Proinflammatory cytokines like IL-1, IL-6, TNF-alpha and growth factor vascular endothelial growth factor (VEGF) as well as anti-inflammatory cytokine IL-10 and related molecules have been described in CNS and plasma of experimental models of seizures and clinical cases of epilepsy. There are reports suggesting more predispositions to seizures during inflammatory conditions like colitis, pneumonia and rheumatoid arthritis. These inflammatory cytokines and growth factors are also known to have dual roles in affecting seizure susceptibility. It remains to be seen if cytokine modulators can be therapeutically exploited for patients with inflammatory disorder and suffering from epilepsy.

The radiosensitizing effect of a plant withanolide, withaferin A, on the B16F1 mouse melanoma was studied in vivo. Treatment of 100 mm3 tumours with 10 to 60 mg/kg withaferin A intraperitoneally produced a dose dependent increase in growth delay and volume doubling time. Injection of 30-50 mg/kg withaferin A, followed by 30 Gy local gamma irradiation, significantly enhanced the tumour response. No systemic or local adverse reactions were noted in these groups. The drug was most effective when injected intraperitoneally 1 h before irradiation. However, neither the individual agents nor their combination could produce any complete response (tumour cure). Melanoma is a relatively radioresistant tumour. The present results indicate that the radiation response of this tumour can be significantly enhanced by pretreatment with withaferin A.

Relationship between body size and bioaccumulation of copper, zinc, lead and cadmium in the fouling bivalve, M. sallei (Recluz) in Visakhapatnam harbour was studied. While concentration of copper, zinc and lead decrease with increasing size, no such relationship is observed for cadmium.

Experiments were performed in weaning rats to understand the influence of thiocyanate, an hydrolytic product of glucosinolates present in foods, on the generation of T3 in situ by type II 5'deiodinase and the binding of 125I T3 to specific nuclear receptors in developing brain. Feeding of thiocyanate through gestation and lactation resulted in an increase in type II 5'deiodinase activity in cerebrum, cerebellum and brainstem of the 21 day old pups compared to controls. Hypothyroidism induced by thiocyanate further resulted in augmentation of the maximum binding capacity of receptors in the cerebrum of the weaning pups. Affinity constants for binding of 125I T3 were however, unaltered. Increase in type II 5'deiodinase activity and the number of binding sites point to an adaptive increase in response to thiocyanate induced hypothyroidism to maintain the cellular T3 levels within a narrow limit.

The effect of plumbagin, a naphthoquinone from the roots of the Indian medicinal plant Plumbago rosea, and Cobalt-60 gamma radiation was studied on Ehrlich ascites carcinoma in vivo, taking cytogenetic damage and cell cycle changes as experimental endpoints. Plumbagin (5 mg/kg body wt, P1) administered intraperitoneally produced a significant increase in the percentage of S-phase as well as G2-M cells with a corresponding decrease in the G1 phase at different post-treatment times. Radiation (7.5 Gy, RT) alone produced the classical G2 block at 1 hr, which persisted with a continuous increase throughout the post-treatment observation period. The combination treatment produced a similar effect as that of RT on G2-M cells, but its effect on the G1 phase was more pronounced than the latter. While P1 treatment produced a small increase in the percentage of labeled S-phase cells, combination treatment significantly reduced the labeled S-phase cells with a corresponding increase in the unlabeled fraction. Drug or radiation alone significantly increased micronuclei induction at various post-treatment times and the combination of the two further enhanced this effect additively. The mechanism of interaction of P1 with radiation in bringing about this effect is not clear.

Role of transition metal ions in expression of benzene toxicity has been suggested. Intraperitoneal administration of benzene to female albino rats daily for 10 days resulted in accumulation of iron in liver nuclei, without any change in copper content. Incubation of hydroquinone (HQ), one of the principal metabolites of benzene with rat liver nuclei resulted in formation of thiobarbituric acid reactive products (TBAR). However, presence of bathocuproine, a copper chelator and EDTA, an iron chelator caused significant inhibition of TBAR release. Thus, the present study revealed that iron accumulation and involvement of copper in nuclear damage induced by HQ.

Effect of pre- and post-irradiation hyperthermia on the induction of gene conversion (non-reciprocal recombination) in diploid yeast cells was investigated. Post irradiation heat treatment does not significantly modify the frequency of gene conversion. Both low-level (51 degrees C-10 min) and lethal (51 degrees C, 40 min) pre irradiation heat treatments enhanced the gene conversion frequency by 17-49%. There was no quantitative correlation between the observed enhancement and the radiation dose. These results suggest that hyperthermia given prior to radiation not only sensitizes the cells to killing but also increases their convertogenic response. It has been suggested that recombination may be involved in carcinogenesis and tumour promotion. Based on these observations along with other reports, wherein hyperthermia was shown to modify the carcinogenic effects in animals as well as the genetic effects of radiation in vitro, it is possible to suggest that hyperthermia may have a potential to increase the second primary cancers, especially in long term survivors. Certain differences in the mechanism of interaction between radiation and heat in the pre- irradiation and post irradiation modalities appear likely.

DNA ploidy of plasma cells in bone marrow has been indicated to play a role in treatment response of multiple myeloma. Therefore, a prospective study was done to test this correlation. Univariate DNA flow cytometry was done on 13 proved multiple myeloma patients. Patients aged below 50 years showed hypodiploidy, irrespective of 'S' phase population, where as all patients above 50 years had diploidy or hyperdiploidy, except for one patient. Early stage patients (III) with less than 25% plasma cells in bone marrow were all aneuploids. Patients belonging to advanced clinical stage with more than 60% plasma cells in bone marrow with aneuploidy, especially hyperdiploidy (DI1.15), carried a poor prognosis. It was difficult to correlate the 'S' phase fraction with other parameters from the present data. Further study with BrdU labelling to determine the proliferative status of the 'S' phase cells is needed.

Glutamate or deoxyuridine in the presence of Cu2+ and delta-aminolevulinic acid (ALA) reaction resulted in the formation of thiobarbituric acid reactive compounds (TBAR), which was linear up to 2 hr. TBAR increased linearly with the increase of both ALA and copper ion concentration. Oxygen consumption was directly proportional to the concentration of formaldehyde perhaps formed by secondary oxidation of dimethyl sulfoxide. Addition of oxyradical scavengers in the reaction mixture resulted in the inhibition of TBAR formation.

Mouse toxicity due to microcystin (hepatotoxin) was detected in water bloom of a local lake, comprising mainly Microcystis aeruginosa. Both toxicity and biomass content of the phytoplankton varied seasonally. Incubations with an algicidal metabolite from Oscillatoria late-virens and a herbicide 3-(3,4-dichlorophenyl)-1, 1-dimethyl urea (DCMU) caused depigmentation and detoxification in bloom samples. Algicide-incubation also encouraged growth of protozoa and bacteria, pathogenic form included.

IDPH-8261, methyl alpha-methyl-4-(3-thienyl)benzeneacetate, exhibited marked anti-inflammatory activity in acute, subacute and chronic models of inflammation. In rats, IDPH-8261 exhibited a dose related inhibition of carrageenin-induced rat paw edema and the inhibition was greater than ibuprofen, phenylbutazone, but was three times less than indomethacin. It exhibited anti-inflammatory activity in normal and adrenalectomized rats. It also exhibited the activity against various phlogistic agents. IDPH-8261 exhibited AI activity in subacute granuloma tests. In adjuvant-induced established polyarthritis. IDPH-8261 exhibited anti-arthritic effect at a very low dose (ED50 = 4 mg/kg, p.o.). Ulcerogenic liability was the lowest (UD50 = 180 mg/kg, p.o.), when compared to reference standard drugs. Low toxicity and high efficacy may make this compound a potentially useful therapeutic agent.

Effect of Septilin, an ayurvedic formulation proven to be effective in the therapy of chronic infections, was investigated on the phagocytic system and humoral response in rats and mice. Septilin exhibited significant protection in E. coli-induced abdominal sepsis in normal mice and in Staphylococcus aureus-induced sepsis in neutropenic mice. It significantly reduced the viable E. coli cells when incubated with neutrophils in rats. Septilin stimulated the phagocytic function of the reticuloendothelial system in mice. In normal rats, Septilin enhanced anti-SRBC hemagglutination antibody titre by 5.7-fold and showed significant protection in cyclophosphamide-induced humoral suppression.

Administration of benzene (ip, 0.5 ml/kg body wt or sc, 1 ml/kg body wt) consecutively for 10 days to male and female rats resulted in decrease in antioxidant potentials in serum. Serum uric acid and albumin showed significant decrease in all groups exposed to benzene. alpha-Tocopherol levels did not exhibit significant change in any of the groups when compared to control. Increase in liver lipid peroxidation and decrease in content of free sulphydryl group were observed in rats exposed to benzene. Serum ferroxidase activity, total iron content (TIC) and total iron binding capacity (TIBC) in female rats exposed to benzene showed significant decrease in ferroxidase activity without any change in TIC or TIBC when compared to control. The decrease in antioxidant potentials observed may be due to oxidation reactions exhibited by benzene metabolites, particularly, hydroquinone and 1, 2, 4-benzenetriol, resulting in oxidative stress in treated rats.

Subchronic oral exposure to styrene in rodents (25 or 50 mg/kg/day in mice; 160 or 320 mg/kg/day in rats and guinea pigs, 5 days/week) for 4 weeks resulted in moderate congestion of pancreatic lobules, focal inflammatory reactions around islets (in mice) and altered serum insulin level while blood glucose levels remained unaffected. Increased beta cell degranulation together with characteristic neoformation of islets were predominantly seen in pancreas of guinea pigs.

Median lethal dose (LD50) of undiluted liquid insect repellent N,N-diethylphenylacetamide (DEPA) in male mice, rats and rabbits was 900, 825 and 635 mg/kg respectively when administered by gavage. Signs of DEPA intoxication point to stimulation of central nervous system (CNS). Acetazolamide (10 mg/kg), sodium bicarbonate (40 mg/kg), and atropine (5 mg/kg) when injected (ip) 5 min after a lethal oral dose of DEPA (1700 mg/kg) did not prevent mortality, while sodium pentobarbital (SPB; 20 mg/kg) when injected 5 min after or 15 min before DEPA provided greater protection to the animals. SPB pretreatment elevated the LD50 of DEPA to 1780 and 1535 mg/kg in mice and rats respectively and 85% rats survived when SPB was injected 5 min after acute oral exposure to DEPA (1000 mg/kg). Carboxylesterase (CaE) inhibition is not a factor in the protection mechanism of SPB. DEPA (1000 mg/kg) when given orally elevated blood PCO2 and reduced pH, O2 content and per cent O2 saturation, while administration of SPB after the same dose of DEPA reduced the degree of acidosis and raised PCO2, and increased the O2 content and per cent O2 saturation to near normal status. The CNS depressant action of SPB may be a crucial factor in protection of rats from DEPA poisoning.

Halotolerant fungus, A. repens, showed a considerable difference in its growth rate, morphology, ultrastructural and molecular composition under NaCl stress as compared to control i.e. non-stressed condition. Light microscopic observations revealed significant differences in their mycelial thickness, their branching and septa. Transmission electron microscopic observations of both the conditions depicted significant differences in the qualitative and quantitative changes in mitochondria. Frequent pinocytotic vesiculation (vacuoles) of plasma membrane was observed in fungus under stress but no such vesiculation in control. The multivesiculate structures observed under stress with their origin from the cell membranes and subsequent release into vacuoles have not been reported in fungi under normal physiological conditions. The observations on pinocytosis are discussed in relation to ion compartmentation and salt tolerance in A. repens.

Oral administration of trichloroethylene (TCE; 0, 500, 1000 and 2000 mg/kg/day) to male mice once daily, 5 days a week for a period of 28 days, caused a significant increase in liver weight, degeneration/necrosis of hepatocytes and characteristics proliferation of endothelial cells of hepatic sinusoids. Increase in kidney weight, glomerular nephrosis, degeneration/desquamation of tubular epithelium and characteristic amyloid deposition in glomeruli were observed only in the group of mice treated with 2000 mg/kg TCE. These changes occurred concurrently with a significant increase in total protein and free sulphydryl contents, elevated activities of acid phosphatase and catalase and decreased activity of delta-aminolevulinic acid dehydratase (delta-ALAD) indicating the sensitivity of liver and kidney as target tissues in TCE-toxicity. Hematological studies showed a significant increase in RBC counts and a reduction in WBC counts without any statistically significant change in the hemoglobin, urea nitrogen, creatinine and uric acid levels in the blood of TCE-exposed mice. A dose-related increase in cell density and acid phosphatase activity with a parallel significant decrease in the activity of delta-ALAD were observed in the bone marrow, which appear to be responsible for hematological alterations in TCE-exposed mice. The results suggest that early metabolic, pathological and hematological perturbations following a short-term exposure of TCE in mice, can provide the basis for its documented potential for chronic effects like blood dyscrasia and cancer.

Carrageenin (2%) was used to produce edema and hyperalgesia; indomethacin, phenylbutazone, aspirin, ibuprofen, analgin, paracetamol and phenacetin were tested at different doses for anti-inflammatory and analgesic activity in the same rats as the peak for the edema reached at the end of 3rd hr and for the hyperalgesia at the end of 4th hr. Indomethacin, phenylbutazone and ibuprofen reduced edema and increased the pain threshold. Analgin and aspirin increased the pain threshold relatively at a low dose. Paracetamol and phenacetin were inactive in the doses tested. Carrageenin (2%) was observed to possess both phlogistic and allogenic properties.

Valproic acid in 100, 200 and 400 mg/kg doses produced a significant dose dependent decrease in exploratory behaviour, tested as number of head dips on the hole board. In the open field test, control mice entered less number of peripheral squares and more number of central squares on day 4 as compared to day 1 of the test. In the lower doses (100 and 200 mg/kg) valproic acid increased central square entries on day 1 with significant decrease by all doses on subsequent days indicating inhibition of exploratory behaviour. However, in peripheral square entry they followed the same pattern as control mice. Neither carbamazepine (10 and 20 mg/kg) nor ethosuccimide (100 and 200 mg/kg) affected exploratory behaviour in either the hole board or open field test.