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Biomedical subjects

S N Upadhyay

Publications and source records attributed to S N Upadhyay.

At least 19 recordsLinked to original sources

Optimization of nutrients for gellan gum production by Sphingomonas paucimobilis ATCC-31461 in molasses based medium using response surface methodology.

A molasses based medium for the production of gellan by Sphingomonas paucimobilis ATCC-31461 was developed. Placket-Burman design criterion was applied to study the effect of various nutrient supplements on gellan production using molasses. Among the 20 variables tested, molasses, tryptone, casaminoacid, disodium hydrogen orthophosphate and manganese chloride showed significant effect on gellan production. A central composite design was applied to determine the optimum concentrations of the significant variables obtained from Placket-Burman design. Most suitable medium composition for production of gellan was (g/l): molasses-112.5; tryptone-1; casaminoacid-1; disodium hydrogen orthophosphate-1; manganese chloride-0.947 and the optimum gellan production was 13.814 g/l.

Culture Media↗

Removal of toluene vapour using agro-waste as biofilter media.

Biodegradation of toluene vapour was investigated in a laboratory scale biofilter packed with cylindrical pieces of yellow-gram (Cajanus cajan) stalk. Inlet concentrations and volumetric flow rates of toluene were varied from 2.56 to 34.73 g/m3 and 0.18 to 0.24 m3/h, respectively. The steady state was achieved within seven days and the degradation of toluene followed an exponential behaviour with time. Elimination capacity increased and tended towards a constant value but removal efficiency decreased with increase in inlet toluene loading. Depending upon loading rate, the process was either mass transfer or reaction-controlled.

Air Pollutants↗

Performance evaluation of an agro waste based biofilter treating toluene vapours.

Biodegradation of toluene vapour was investigated for 162 days in a laboratory scale biofilter packed with maize waste. Inlet concentration of toluene was varied from 2.86 to 5.79 g m(-3) and the volumetric flow rate of toluene loaded air from 0.09 to 0.3 m3 h(-1). The media bed height used in the study was 0.63 m. Plug flow behaviour of the bed was confirmed by RTD experiments. The steady state was achieved in 18 days. Depending upon initial loading rates, removal efficiency ranging from 54.5 to 85.3% and elimination capacity ranging from 43.47 to 174.6 g h(-1) m(-3) were observed during steady state operation. The dynamic behaviour of the biofilter was evident from response to change in process conditions. The stability of the biomass was evident from the fast response of the biofilter to shutdown and restart operations. High value of carbon recovery confirms the effective biodegradation in the biofilter. In the startup period, the pressure drop in the bed increased for some time after that it tended towards a constant value.

Air Pollutants↗

Studies on the immunogenic potential of plant-expressed cholera toxin B subunit.

Nicotiana tabacum var. Samsun was transformed via Agrobacterium-mediated transformation with a gene encoding the cholera toxin B subunit (CTB) of Vibrio cholerae, modified to contain a sequence coding for an endoplasmic reticulum retention signal (SEKDEL), under the control of the cauliflower mosaic virus 35S promoter. Total protein from the transgenic leaf tissue was isolated and an aliquot containing 5 microg recombinant CTB was injected intradermally into Balb/c (H2K(d)) mice. CTB-specific serum IgG was detected in animals that had been administered plant-expressed or native purified CTB. A T-cell proliferation study using splenocytes and cytokine estimations in supernatants generated by in vitro stimulation of macrophages isolated from the immuno-primed animals was carried out. Inhibition of proliferation of T lymphocytes was observed in splenic T lymphocytes isolated from animals injected with either native or plant-expressed CTB. Macrophages isolated from mice immunised with native or plant-expressed CTB showed enhanced secretion of interleukin-10 but secretion of lipopolysaccharide-induced interleukin-12 and tumor necrosis factor alpha was inhibited. These studies suggest that plant-expressed protein behaved like native CTB with regards to effects on T-cell proliferation and cytokine levels, indicating the suitability of plant expression systems for the production of bacterial antigens, which could be used as edible vaccine. The transgene was found to be inherited in the progeny and was expressed to yield a pentameric form of CTB as evident by its interaction with G(M1) ganglioside.

Animals↗

Microbial corrosion monitoring by an amperometric microbial biosensor developed using whole cell of Pseudomonas sp.

A microbial biosensor was developed for monitoring microbiologically influenced corrosion (MIC) of metallic materials in industrial systems. The Pseudomonas sp. isolated from corroded metal surface was immobilized on acetylcellulose membrane and its respiratory activity was estimated by measuring oxygen consumption. The microbial biosensor was used for the measurement of sulfuric acid in a batch culture medium contaminated by microorganisms. A linear relationship between the microbial sensor response and the concentration of sulfuric acid was observed. The response time of biosensor was 5 min and was dependent on the immobilized cell loading of Pseudomonas sp., pH, temperature and corrosive environments. The microbial biosensor response was stable, reproducible and specific for sensing of sulfur oxidizing bacterial activity.

Biosensing Techniques↗

In vitro propagation of Withania somnifera and isolation of withanolides with immunosuppressive activity.

Withania somnifera plantlets were produced in vitro from the shoot-tip of aseptically germinated seedlings. Culture conditions were optimized using different plant growth regulators which gave rise to 120 shoots from a single bud. The plantlets were then transferred to pots and maintained in greenhouse for 4 months. 90% of these in vitro propagated plantlets survived and showed normal growth. Leaves from these plants were used for isolation of the withanolides. Methanolic extract of leaves from plantlets growing in tissue culture and those transferred to the greenhouse were evaluated for immunomodulatory activity. While the extract from greenhouse samples showed potent immunosuppressive activity, those from tissue cultures samples did not show any activity. Fractionation and characterization of withanolides, using HPLC, NMR, MS methods revealed the presence of withaferin A in the greenhouse samples. Our results indicate that Withania species may require longer time and better differentiation and also natural environment for the production of withaferin A.

Adjuvants, Immunologic↗

Effect of gamma-irradiation on H3 histone and DNA in solution.

Three methods, namely, absorbance of colour by reaction with Folin-Ciocalteau reagent, UV absorbance and fluorescence intensity measurements for detection of H3 histone in 0.15 M standard saline citrate (SSC) solution were compared. Maximum sensitivity was found with the Folin-Ciocalteau method. Effect of varying pH and of gamma- radiation on H3 histone and on interaction of H3 histone with DNA were studied. For this, solutions of H3 histone in SSC, in 0.9% NaCl, H3 histone + DNA in 0.9% NaCl were subjected to varying pH (1-10) and gamma- radiation (dose 10-50 Gy) and lambda(max) and Alambda(max) were monitored. From the molar ratios of histone and DNA in the complex, it was observed that at gamma -radiation dose of 50 Gy and pH 8.54, there was a depletion of 6-8 microg/ml of histone from the histone-DNA complex.

Animals↗

Comparative effects of prenatal and postnatal undernutrition on learning and memory in rats.

Effects of pre- and post-natal undernutrition on learning and memory parameters were studied in albino rats. Prenatal undernutrition was induced in rat pups by restricting the mother's diet by 50% during the entire gestation period, whereas postnatal undernutrition was induced in rat pups by restriction of their diet by rotating them between lactating and non-lactating maternalised females for 12 hr each day during suckling period from 2nd day to 18th day after birth. At 2.5 to 3 months of age all the rat offsprings were subjected to (i) original and reversal discrimination learning, (ii) passive avoidance, and (iii) active avoidance and its retention tests. The results indicate that both pre- and post-natal undernutrition in rat pups caused significant deficits in original and reversal discrimination learning, retention of passive avoidance after one week retention interval, and retention of active of avoidance learning. However, both pre- and post-natal undernutrition did not show significant effect on acquisition of active avoidance and retention of passive avoidance after 24 hr retention interval.

Animals↗

Immunocontraceptive activity guided fractionation and characterization of active constituents of neem (Azadirachta indica) seed extracts.

A novel approach for immunocontraception by intervention of local cell mediated immunity in the reproductive system by using single intrauterine application of neem oil has been described earlier. The reversible block in fertility was reported to last for 107-180 days in female Wistar rats (Upadhyay et al., 1990. Antifertility effects of neem oil by single intrauterine administration: A novel method of contraception. Proceedings Of The Royal Society Of London B 242, 175-180) and 7-11 months in monkeys (Upadhyay et al., 1994. Long term contraceptive effects of intrauterine neem treatment (IUNT) in bonnet monkeys: An alternative to intrauterine contraceptive devices. Contraception 49, 161-167). The present study, describes the identification and characterization of the biologically active fraction from neem seeds (Azadirachta indica A. Juss. Family Meliaceae), responsible for the above activity in adult female Wistar rats. Initial studies with the mechanically extracted oil and solvent extracts of neem seeds have revealed that the antifertility activity was present in constituents of low to intermediate polarity. A hexane extract of neem seeds was reported to be biologically active (Garg et al., 1994. Comparison of extraction procedures on the immunocontraceptive activity of neem seed extracts. Journal of Ethnopharmacology 22, 87-92). Subsequently, hexane extract was sequentially fractionated through the last active fraction using various separation techniques and tested for antifertility activity at each step. Preparative HPLC was used for isolating individual components of the active fraction in quantities, sufficient for characterization. An analytical HPLC method was developed for standardization of the fraction. The active fraction was identified to be a mixture of six components, which comprises of saturated, mono and di-unsaturated free fatty acids and their methyl esters. Dose response study was performed with the last active fractions. The antifertility activity with the active fraction was reversible in nature and it was completely active until 5% concentration. There was no systemic toxic effect following the administration of the active fraction. This study, for the first time, proposes an active fraction from neem seeds, responsible for long term and reversible blocking of fertility after a single intrauterine administration with high efficacy.

Animals↗

Radioprotective efficacy of HT + AET in encapsulated form.

Entrapment of 5-hydroxyl-L-tryptophan (HT) in erythrocyte ghost prepared by hypotonic method and high voltage electric discharge method are nearly same. Release of HT with beta-aminoethylisothiuronium bromide hydrobromide (HT + AET) in in vitro system is rapid but only a portion of the entrapped amount is released. Release of HT + AET in serum marginally increases at 2 hr. Compared to release in in vitro medium the release in serum is less. Survival studies with Swiss albino mice indicates that compared to HT alone, the combination of HT + AET shows about 9 times percentage survival. The same combination in the encapsulated form show comparable percentage survival though the amount needed is 1/200th times compared to free form.

Animals↗

Behavioural effects of prenatal and postnatal undernutrition in rats.

Effects of pre- and post-natal undernutrition on anxiety and depression paradigms were studied in albino rats. Prenatal undernutrition was induced in rat pups by restricting the dam's daily food during the gestation period whereas postnatal undernutrition in rat pups was induced by rotating them between lactating and non-lactating maternalised females daily for 12 hr during suckling period from 2nd to 18th day after birth. At 2.5 to 3 months of age all the rat pups were subjected to (i) elevated plus maze behaviour, (ii) open-field behaviour, and (iii) swimming induced behavioural despair tests. The results indicate that postnatal undernutrition caused significantly increased anxiety in the elevated plus maze as well as in open-field behaviour tests. Whereas prenatal undernourishment caused lesser degree of anxiogenic behaviours in the elevated plus maze test. Prenatally undernourished rats showed increased anxiety in the open-field behaviour test. Both, pre- and post-natal undernutrition also lead to increased depressive behaviour in the behavioural despair test and postnatal undernourishment caused greater degree of behavioural despair.

Animal Nutritional Physiological Phenomena↗

Safety of intrauterine administration of purified neem seed oil (Praneem Vilci) in women & effect of its co-administration with the heterospecies dimer birth control vaccine on antibody response to human chorionic gonadotropin.

Praneem Vilci (PV), purified neem oil was reported to exercise a reversible antifertility effect after a single intrauterine instillation in rodents and primates without any adverse effects. After toxicology, drug regulatory and ethical clearances, a phase I clinical trial was conducted on PV. Eighteen healthy tubectomised women were enrolled to evaluate the safety of a single intrauterine instillation of PV and to determine the effect of its co-administration on anti-hCG response to the heterospecies dimer (HSD) hCG vaccine. Eight women received PV alone and ten women were given the HSD-hCG vaccine in addition. Base-line and post-treatment haematological and biochemical profiles were determined as also the mid-luteal serum progesterone. Endometrial biopsies were examined to assess ovulatory status and the effect of intrauterine treatment with PV on the endometrium. Anti-hCG antibody titres were estimated in women who were concurrently immunized with the HSD vaccine. No untoward reaction was observed in any woman. Menstrual pattern and ovulatory status remained unaltered. Endometrial biopsy after PV instillation in one woman showed non-specific endometritis but she remained asymptomatic. Mild eosinophilia was seen in two women and this reverted to normal on its own. All women receiving PV and the HSD vaccine generated antibodies against hCG. Our data show that intrauterine administration of PV is safe and does not prevent the antibody response to HSD-hCG vaccine.

Adult↗

Synergistic spermicidal activity of neem seed extract, reetha saponins and quinine hydrochloride.

In order to identify potent spermicidal agents which are free from the side effects of currently available agents, spermicidal activity of purified neem seeds extract (Praneem), reetha saponins and quinine hydrochloride was studied individually and in combination. Sander-Cramer test was used to assess the activity on human sperm. Under the test conditions, minimum effective spermicidal concentrations for Praneem, reetha saponins and quinine hydrochloride were 25%, 0.05% and 0.346%, respectively. At these concentrations, 100% of the sperm were immobilised within 20 seconds. A positive synergistic effect in the spermicidal activity of these components, if used in combination, was observed which implies the use of reduced concentrations of each to bring about the desired action. The selected combination formulated into a suitable dosage form is likely to offer dual benefit of a potent contraceptive and an antimicrobial preparation.

Drug Combinations↗

Comparison of extraction procedures on the immunocontraceptive activity of neem seed extracts.

Azadirachta indica (Neem) seed extracts are known to activate the local cell-mediated immune reactions after a single intrauterine administration, leading to a long term reversible block of fertility. In order to identify and characterize the active fraction responsible for this activity, neem seeds were extracted by both mechanical expression and solvent extraction using a range of polar to non-polar solvents which yielded 3 broad fractions. The mechanically expressed oil was fractionated using different approaches and studied for antifertility activity. The hexane extract and a corresponding column fraction showed potent and reproducible antifertility activity. Other fractions were less stable with regard to reproducibility of effects and composition. It is our conclusion that for subsequent fractionation to reach the last active fraction, the hexane extract is the most useful starting material.

Animals↗

Studies on the contraceptive efficacy of Praneem polyherbal cream.

Praneem polyherbal cream, a spermicidal formulation, has been developed at the National Institute of Immunology, which makes use of Praneem, a purified extract from the dried seeds of an ancient Indian plant Azadirachta indica (Neem), extract from the pericarp of fruits of Sapindus species and quinine hydrochloride. These ingredients have a synergistic spermicidal activity and an optimised formula was derived. The components were made into a water-soluble cream base prepared by using pharmaceutically acceptable base and stabilised by addition of IP grade antioxidant and preservatives. The cream is devoid of irritation and sensitization potential, as seen with standard Draize test on normal and abraded skin of rabbits and by 21-day cumulative skin sensitivity in human volunteers. The formulation was found to be safe under subacute toxicity studies in monkeys. The formulation has shown high contraceptive efficacy in rabbits and in monkeys after intravaginal application. The shelf-life of the cream at room temperature is estimated to be 18 months by accelerated stability studies.

Animals↗

Cellular immune responsiveness in rabbits with Setaria digitata filarial antigen and TDM adjuvant.

The purified surface antigens of the bovine filarial parasite Setaria digitata were used as an antigen to immunize rabbits. The aqueous suspensions of trehalose 6-6' dimycolate (TDM) has been successfully used as an effective immunomodulator in experimental studies on filariasis. The effectiveness of such an antigen-TDM combination was demonstrated by enhanced humoral and cellular immunity. Administration of antigen alone shows only humoral immunity. The detectable cellular immune responses further confirm the effect of filarial antigen-TDM combination. The cell-mediated immunity was expressed in vivo by delayed skin reaction and in vitro by leukocyte and macrophage migration inhibition tests.

Adjuvants, Immunologic↗