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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

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

A birth control vaccine is on the horizon for family planning.

Vaccines for control of fertility are likely to have an important impact on family planning methods. They are designed to act by mobilization of an internal physiological process and do not require external medication on a continuous basis. A number of birth control vaccines are at different stages of development, the most advanced being a vaccine inducing antibodies against human chorionic gonadotrophin (hCG). This vaccine consists of a heterospecies dimer (HSD, beta hCG associated with alpha-subunit of ovine luteinizing hormone, beta hCG:alpha oLH) linked to tetanus toxoid (TT) or diphtheria toxoid (DT) as carriers. The vaccine has recently passed an important milestone; it has completed the first leg of phase II efficacy trials. Women of proven fertility leading active sexual life were protected from becoming pregnant at antibody titres > or = 50 ng of hCG bioneutralization capacity per ml. This vaccine has previously been demonstrated to be reversible in its effect. It is free from any notable side-effects on endocrine, cardiovascular and other body functions. Ovulation was not disturbed and menstrual regularity was maintained. A logistic disadvantage of the present vaccine is the requirement for multiple injections. This is expected to be overcome by encapsulation of the requisite doses of the vaccine in biodegradable microspheres, which could be given at a single contact point for sustained antibody titres lasting over a year. A live recombinant vaccine has also been made that elicits high anti-hCG titres in monkeys for nearly 2 years following primary immunization and a booster at 8-9 months.

Adult

Anxiolytic activity of piracetam, a nootropic agent, following subchronic administration in rodents.

Piracetam (PIR), a cyclic GABA derivative without GABA-mimetic activity, is classified as a nootropic agent, a new class of psychotropic drugs which augment learning acquisition and retention of memory. The present study indicates that PIR has significant anxiolytic activity in rodents following subchronic, but not acute administration, when tested against several paradigms of experimental anxiety. Thus, PIR (250 and 500 mg/kg), administered orally for 7 and 14 days, exhibited anxiolytic activity in the open-field, elevated plus-maze and footshock-induced fighting in paired mice paradigms, as well as in the Vogel's conflict test in rats. In addition, PIR induced significant reduction in rat brain tribulin levels, a putative endocoid marker for anxiety, produced by pentylenetetrazole, an anxiogenic agent. On the contrary, single acute administration of PIR failed to induce any anxiolytic effect. The present study, thus, confirms clinical reports that PIR can induce a delayed antianxiety effect in psychogeriatric individuals and in chronic alcoholism.

Animals

Latency of memory consolidation induced in mice by piracetam, a nootropic agent.

Piracetam (PIR), a cyclic GABA derivative, is the prototype of a new class of psychoactive drugs, the nootropic agents, which improve learning acquisition and the retention of the learning as memory. It was proposed that nootropics act on processes essentially involved in information storage, thus facilitating memory. This property can best be investigated by drug administration after the learning trial and assessing subsequent retention performance. The present study was designed to evaluate the effective time period of memory consolidation, induced by piracetam, by assessing the retention of a learned task in two behavioural paradigms, following administration of the drug after learning acquisition. Physostigmine was used as the standard drug because of its well established facilitation of memory storage. PIR (250 and 500 mg/kg, ip) and physostigmine (0.05 mg/kg, ip) were administered in different groups of mice 5 min, 1, 2, 4, 8, 12 and 24 hr after learning acquistion of two passive avoidance tasks and the retention performance was evaluated 3 days later. The results indicate that, while physostigmine induced significant memory consolidation when administered up to 2 hr after learning acquisition, PIR induced retention of learning beyond this period. Thus, the highest effective post-trial interval for the lower and higher dose of the drug was 8 and 12 hr, respectively, in both the test paradigms. The results confirm that nootropics, like piracetam, are capable of memory consolidation, as assessed by retention of learning, even after intervals of 8 to 12 hr between learning and drug treatment.

Animals

Effect of piracetam on electroshock induced amnesia and decrease in brain acetylcholine in rats.

Piracetam, a prototype of a new class of psychotropic agents, the nootropic agents, which improve learning ability and memory retention, was found to induce a dose-related prevention of disruption of acquisition of a passive avoidance response produced by electroshock application. The amnesia attenuating effect of piracetam was accompanied by prevention of the decrease in acetylcholine concentrations of rat brain induced by electroshock. The study indicates that the cognition enhancing effect of piracetam may be due to a facilitatory effect on cholinergic transmission.

Acetylcholine

Immunomodulatory effects of neem (Azadirachta indica) oil.

Immunomodulatory effects of neem oil were studied in mice. The animals were treated intraperitoneally (i.p.) with neem oil; control animals received the emulsifying agent with or without peanut oil. Peritoneal lavage, collected on subsequent days, showed a maximum number of leukocytic cells on day 3 following treatment with neem oil; peritoneal macrophages exhibited enhanced phagocytic activity and expression of MHC class-II antigens. Neem oil treatment also induced the production of gamma interferon. Spleen cells of neem oil-treated animals showed a significantly higher lymphocyte proliferative response to in vitro challenge with Con A or tetanus toxoid (TT) than that of the controls. Pre-treatment with neem oil, however, did not augment the anti-TT antibody response. The results of this study indicate that neem oil acts as a non-specific immunostimulant and that it selectively activates the cell-mediated immune (CMI) mechanisms to elicit an enhanced response to subsequent mitogenic or antigenic challenge.

Adjuvants, Immunologic

Effect of dihydroergotoxine, a cerebral vasodilator, on cognitive deficits induced by prenatal undernutrition and environmental impoverishment in young rats.

The study was conducted on 64 Charles Foster strain albino rats, which were equally distributed into 8 evenly matched groups, following a 2 x 2 x 2 factorial design, by varying three independent factors at two levels: nutrition--normal and undernutrition; environment--enrichment and impoverishment, and drug treatment--vehicle and dihydroergotoxine (3 mg/kg, i.p.). Prenatal undernutrition was induced by restricting the mother's food intake. The environmental enrichment/impoverishment and the vehicle/dihydroergotoxine treatments were given during the postweaning period of the pups. The rats were subjected to original and subsequent reversal brightness discrimination learning tests in a single unit T-maze at 8-9 weeks of age. Thereafter, the animals were tested for passive avoidance learning. The results indicate that undernutrition caused significant original and reversal discrimination learning, deficits whereas environmental deprivation attenuated only the original discrimination learning performance. Dihydroergotoxine treatment facilitated the learning performance of rats in both the original and reversal learning tests. Nutritional, environmental and dihydroergotoxine treatments had no effect on the retention of the passive avoidance learning, both at 24 hr and 1 week intervals. Dihydroergotoxine treatment attenuated the learning deficits induced by prenatal undernutrition. The results indicate that dihydroergotoxine is not likely to be useful in cognitive deficits, induced by malnutrition, though it facilitated learning acquisition, since it had no effect on retention.

Animals

Antifertility effects of neem (Azadirachta indica) oil by single intrauterine administration: a novel method for contraception.

A novel use of neem (Azadirachta indica) oil, a traditional plant product, for long-term and reversible blocking of fertility after a single intrauterine application is described. Female Wistar rats of proven fertility were given a single dose (100 microliters) of neem oil by intrauterine route; control animals received the same volume of peanut oil. Whereas all control animals became pregnant and delivered normal litters, the rats treated with neem oil remained infertile for variable periods ranging from 107 to 180 days even after repeated matings with males of proven fertility. The block in fertility was, however, reversible as half of the animals regained fertility and delivered normal litters by five months after treatment, without any apparent teratogenic effects. Unilateral administration of neem oil in the uterus blocked pregnancy only on the side of application whereas the contralateral uterine horn treated with peanut oil had normally developing foetuses; no sign of implantation or foetal resorption was noted in the neem-oil-treated horn. The ovaries on both sides had 4-6 corpora lutea indicating no effect of treatment on ovarian functions. The animals treated with neem oil showed a significant leukocytic infiltration in the uterine epithelium between days 3 and 5 post coitum, i.e. during the pre-implantation period. Intrauterine application of neem oil appears to induce a pre-implantation block in fertility; the possible mechanisms of the antifertility action are discussed.

Animals

Effect of pyritinol, a cerebral protector, on learning and memory deficits induced by prenatal undernutrition and environmental impoverishment in young rats.

The study was conducted on 64 CF strain albino rats, which were equally distributed into 8 evenly matched groups following a 2 x 2 x 2 factorial design, by varying three independent factors at two levels: nutrition--normal and undernutrition; environment--enrichment and impoverishment, and drug treatment--vehicle and pyritinol (100 mg/kg, ip). Prenatal undernutrition was induced by restricting the mother's food intake. The environmental enrichment/impoverishment and the vehicle/pyritinol treatments were given during the postweaning period of the pups. The rats were subjected to original and subsequent reversal brightness discrimination learning tests in a single unit T-maze at 8-9 weeks of age. Thereafter, the animals were tested for the passive avoidance learning. The results indicate that undernutrition caused significant original discrimination learning deficits whereas environmental deprivation attenuated both the original and reversal learning performance. Environmental impoverishment attenuated the retention of passive avoidance behaviour but undernutrition had no effect on this paradigm. Pyritinol treatment improved the learning and retention performance of normally reared rats and also attenuated the original and reversal learning deficits induced by parental undernutrition and postweaning environmental impoverishment. The results indicate that pyritinol may be useful in learning and memory deficits induced by malnutrition and environmental deprivation.

Animals

Functional morphology of testis and its excurrent ducts in rats immunized with synthetic luteinizing hormone releasing hormone conjugated to tetanus toxoid.

The effects of active immunization against luteinizing hormone releasing hormone (LHRH) on the testis and its excurrent ducts in rats were studied using a synthetic LHRH peptide conjugated to tetanus toxoid (TT), with or without an adjuvant. The study demonstrated that LHRH-TT given alone did not elicit an appreciable immune response and that its immunogenicity could be enhanced by administration with an adjuvant, such as species-specific IgG or SPLPS. The anti-LHRH antibody response showed a corresponding regressive effect on the morphology of the testis and its excurrent ducts. The functional involution of the testis was reflected in the form of a block in spermatogenesis and the lowering of testosterone production; the basic structural organization of the testicular components was, however, maintained. Epididymal ducts displayed rather drastic atrophic changes, indicating that prolonged immunization may result in irreversible damage to the excurrent ducts and could, therefore, adversely affect the restoration of fertility even after the return of testicular function following decline in the immune response against LHRH. This could be an important consideration if LHRH immunization is to be used for male contraception.

Animals