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Biomedical subjects

R Pal

Publications and source records attributed to R Pal.

At least 73 records · Page 4Linked to original sources
PubMed1993

Immunochemical and immunohistochemical characterization of a synaptic membrane protein that binds the competitive antagonists of NMDA receptors.

An approximately 54-kDa protein that has binding sites for the competitive N-methyl-D-aspartate (NMDA) receptor antagonists 3-((+-)-2-carboxypiperazine-4-yl)-propyl-1-phosphonic acid (CPP) and (+-)-(E)-2-amino-4-propyl-5-phosphonopentenoic acid (CGP 39653) was purified from rat brain synaptic membranes. Polyclonal antibodies to this protein reacted specifically with an approximately 54-kDa protein in synaptic membranes and immunoextracted approximately 60% of [3H]CGP 39653 binding sites associated with solubilized membrane proteins. The antibodies also labeled antigenic sites in the perikaryon and apical and basilar dendrites of pyramidal neurons of the hippocampus and cerebral cortex.

2-Amino-5-phosphonovalerate↗
PubMed1993

Temporal nature of the promoter and not relative strength determines the expression of an extensively processed protein in a baculovirus system.

We demonstrate that the expression of extensively modified and secreted heterologous proteins synthesized in the baculovirus expression vector system (BEVS) depends on the temporal nature of the promoter transcribing the foreign gene. The beta subunit of the human chorionic gonadotropin, an extensively modified secretory glycoprotein hormone was expressed under the transcriptional control of the AcNPV basic protein gene promoter (MP) and the polyhedrin gene promoter (POL), respectively. MP, activated late in the infection cycle, is a weaker promoter when compared to the stronger very late POL promoter. Levels of secretion, immunoreactivity and bioactivity of recombinant proteins, beta hCGMP and beta hCGPOL synthesized under control of the MP and POL promoter were compared. Secretion of beta h CGMP was relatively higher. Enzymatic analysis revealed that the synthesized protein was sialylated. Receptor binding assays and testosterone stimulation assays in a mouse Leydig cell system demonstrated that on a unit protein basis, beta hCGMP was biologically more active than beta hCGPOL.

Animals↗
PubMed1993

Conformational perturbation of the envelope glycoprotein gp120 of human immunodeficiency virus type 1 by soluble CD4 and the lectin succinyl Con A.

We have studied perturbation of the gp120/gp41 envelope complex of HIV-1 in the presence of the mannose-specific lectin succinyl Con A (SC) and compared the effect with that observed in the presence of soluble CD4 (sCD4). SC did not inhibit the binding of gp120 to CD4. Both sCD4 and SC inhibited syncytium formation induced by HIV-1-infected Molt3/HIV-1IIIB cells. The infectivity of HIV-1 was markedly reduced when the virions were preincubated with SC or when SC was mixed simultaneously with virus and cells. The conformation of gp120 was altered in the presence of SC as evidenced by an increased susceptibility of the principal neutralizing epitope (V3 loop) to thrombin digestion. SC treatment of [35S]-methionine-labeled virions derived from Molt3/HIV-1IIIB cells resulted in the dissociation of gp120 from the viral membrane. The effect was less pronounced than that observed with sCD4. These results suggest that although interacting with different regions of gp120, the mannose-specific lectin alters the conformation of the glycoprotein in a manner similar to that induced by sCD4, causing destabilization of the gp120/gp41 complex.

CD4 Antigens↗
PubMed1993

Antibody response against epitopes on hCG mapped by monoclonal antibodies in women immunized with an anti-hCG vaccine and its implications for bioneutralization.

The immunological determinants on hCG, to which an antibody response is generated in women by the contraceptive HSD vaccine, were mapped by using a panel of anti-hCG monoclonal antibodies. Two types of inhibition enzyme immunoassays (EIAs) were used to analyze 126 serum samples from 18 subjects immunized with the vaccine and protected from pregnancy. Monoclonal antibody (MAb) 357-2, which recognizes beta-hCG loop peptide 38-57, did not inhibit significantly the binding of immune sera to hCG. Moreover, none of the sera reacted with this loop peptide in a direct binding EIA, suggesting weak immunogenicity of this epitope. All sera competed with MAb 206 and a preponderance of antibodies was seen against this epitopic region on beta-hCG. The antibody titers against the MAb 206 epitope showed a good correlation with the bioneutralization capacity of the sera throughout the course of immunization. These studies indicate the presence of an immunodominant antigenic determinant on hCG as recognized by the human immune system.

Animals↗
PubMed1993

Influence of ethanol on cadmium accumulation and its impact on lipid peroxidation and membrane bound functional enzymes (Na+, K(+)-ATPase and acetylcholinesterase) in various regions of adult rat brain.

Influence of ethanol on cadmium accumulation and its effect on metallothionein induction, binding of cadmium to metallothionein, lipid peroxidation and membrane bound functional enzymes such as (Na(+)-K+)-ATPase and acetylcholinesterase in various regions of adult rat brain was investigated. Ethanol (2 g/kg body wt) and cadmium (1 mg/kg body wt) were administered alone as well as in combination to different groups of rats, i.p., for a period of 1 week. It was observed that cadmium when co-administered with ethanol led to pronounced increase in cadmium accumulation in various regions of the brain. This ethanol induced accumulation of cadmium did not induce the synthesis of metallothionein and also did not bind to this protein in brain and mainly was present as non-metallothionein bound cadmium. It lead to a significant increase in lipid peroxidation and inhibition of membrane bound functional enzymes; (Na(+)-K+)-ATPase and acetylcholinesterase in various regions of the brain indicating functional impairment. The results of the present study imply that ethanol renders the adult brain more susceptible to cadmium neurotoxicity. Corpus striatum and cerebral cortex are more vulnerable regions than other areas of the brain.

Acetylcholinesterase↗
PubMed1993

Lipid peroxidation and antioxidant defense enzymes in various regions of adult rat brain after co-exposure to cadmium and ethanol.

Effect of cadmium (1 mg/kg body weight) and ethanol (2 g/kg body weight) exposure, alone as well as in combination, on essential trace metal homeostasis, lipid peroxidation and antioxidant defense enzymes in various regions of the adult rat brain was investigated. It was observed that cadmium when administered along with ethanol accumulated significantly in corpus striatum (3.5 fold) and cerebral cortex (3.0 fold) compared to the cadmium treated group. The ethanol induced accumulation of cadmium led to significant depletion in the levels of essential trace metals like zinc and copper in these regions of the brain. Further, cadmium or ethanol alone did not show any significant effect on lipid peroxidation and antioxidant defense enzymes in any of the regions of the adult brain but when given in combination, caused a significant increase in lipid peroxidation and markedly decreased the activities of antioxidant defense enzymes like glutathione peroxidase, superoxide dismutase and catalase particularly in corpus striatum and cerebral cortex. Structural alterations produced by increased lipid peroxidation after cadmium and ethanol co-exposure may have profound effect on the activities of membrane bound enzymes and hence may lead to functional impairment. The results of the present study imply that ethanol renders the adult brain more susceptible to the neurotoxic effects of cadmium. Corpus striatum and cerebral cortex are more vulnerable to the toxic effects of cadmium under the influence of ethanol than other regions of the brain.

Animals↗
PubMed1993

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↗
PubMed1992

Simultaneous synthesis of enzymatically active luciferase and biologically active beta subunit of human chorionic gonadotropin in caterpillars infected with a recombinant baculovirus.

The beta subunit of human chorionic gonadotropin (beta hCG), a secretory and extensively glycosylated hormone, and firefly luciferase, a non-secretory enzyme, were simultaneously synthesized in Spodoptera larvae upon infection with a dual expression recombinant baculovirus, vAc beta hCG-luc. Luciferase was retained predominantly in the body tissue while beta hCG was secreted into the hemolymph of infected larvae. Both the proteins were similar to their authentic counterparts in terms of immunoreactivity and bioactivity. The caterpillar-derived recombinant hCG exhibited reduced electrophoretic mobility on SDS-PAGE and increased biological activity as compared to the hCG expressed in insect cells in culture. The implications of using the larval system for expressing an extensively glycosylated protein are discussed.

Animals↗
PubMed1992

Neurotoxicity of dichlorvos: effect on antioxidant defense system in the rat central nervous system.

The effect of dichlorvos exposure (5 mg kg-1 body wt, ip) on lipid peroxidation and antioxidant defense system in different regions of the rat central nervous system was studied. In the present paper an inhibition of acetylcholinesterase activity was used as an index of dichlorvos neurotoxicity. We observed significant increases in the activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase which were accompanied by a decrease in the values of lipid peroxidation. Dichlorvos exposure also resulted in a significant decrease in glutathione peroxidase activity. The decreased levels of both reduced and oxidized glutathione as observed on dichlorvos exposure affected the GSH/GSSG ratio. These results indicate that the enzymes SOD and catalase may enhance the disposal of potentially toxic radicals. Furthermore, the decrease in GSH levels may be a mechanism for the detoxification of dichlorvos in the brain.

Acetylcholinesterase↗
PubMed1992

Biological actions of monoclonal antibodies to bovine lutropin receptor.

Monoclonal antibodies (Mabs) were produced against bovine Lutropin receptor (LH-R). Antibodies were detected by an enzyme linked immunosorbant assay (ELISA). Hybridomas were subcloned to achieve monoclonality. Ascites were developed in Balb/c mice. Hybridoma supernatants were purified by ammonium sulfate precipitation and chromatography on hydroxylapatite columns. LH-R antibodies showed upto 50% inhibition of 125I-labeled hCG binding to bovine luteal cell membranes and up to 80% inhibition of testosterone production by hCG stimulated mouse Leydig cells. LH-R antibodies were predominantly IgM isotype. Purified antibodies showed a 78-kDa band, in SDS-PAGE, as the heavy chain of the immunoglobulin. LH-R antibodies were localized specifically in the thecal and luteal cells of the rat ovaries as well as in the Leydig cell of mouse testes. Injections of the LH-R antibody caused a constant estrus in normal rats. One month after the cessation of the injections the animals returned to normal estrus cycle and fertility. Pregnant mice injected with LH-R antibodies produced only 3 viable pregnancies and 10 pups, as compared to 8 pregnancies and 45 pups born to normal controls. LH-R antibodies also caused, approximately, a 50% reduction in testosterone production in normal male rats. These observations indicate a high degree of specificity of the Mab to LH-R and their potential in studies on gonadal function.

Animals↗
PubMed1992

Active immunization of baboons (Papio anubis) with the bovine LH receptor.

Four baboons (Papio anubis) were actively immunized with bovine LH receptor for periods of 6-22 months. Serum antibody levels were measured by an enzyme immunoabsorbant assay (ELISA). Antibodies against the receptor were detected 2 weeks after the first injection. Antisera caused an inhibition in the binding of human chorionic gonadotropin (hCG) to its receptor as well as inhibited the production of hCG induced testosterone by rat Leydig cells in culture. Serum estradiol and progesterone levels were determined by radioimmunoassay (RIA). Progesterone levels were suppressed during the post immunization period. Two baboons experienced periods of anovulation. Serum estradiol levels were cyclic and appeared elevated. Baboons were mated with males of proven fertility; none of the immunized females conceived over eight cycles of observation. Fertility parameters returned to normal, when antibodies against LH-receptor were undetectable in the serum.

Animals↗
PubMed1992

Vaccines for control of fertility and hormone dependent cancers.

Two vaccines, namely one inducing antibodies against hCG and the other against GnRH, are now in clinical trials. The hCG vaccine has entered Phase II clinical trials in three centres in India after successfully completing Phase I clinical studies in several centres in India and in four countries abroad. The vaccine was found to be devoid of side-effects; its effect was reversible. The available data on 179 cycles indicate that the vaccine prevents pregnancy at antibody titres above 50 ng/ml. A genetically engineered version of the vaccine has also been approved for trials in human lung cancer patients of the type which make hCG. hCG is observed to be a growth factor for such tumours. The GnRH vaccine is usable in both males and females as the deca-peptide is common to both sexes. Following suitable experimental and toxicology studies, the vaccine is currently in Phase I/Phase II clinical trials in patients of prostate carcinoma. Where antibody GnRH antibodies were induced, the LH, FSH and testosterone levels declined. This was accompanied by a reduction in prostate specific antigen. Clinical improvement was observed in many cases. The vaccine has also entered Phase I clinical studies in postpartum women, with the objective to extend the lactational amenorrhoea and extend inter-child interval.

Chorionic Gonadotropin↗
PubMed1992

Delineation of immunoreactive, conserved regions in the external glycoprotein of the human immunodeficiency virus type 1.

Immunization of mice and rats with purified external glycoprotein gp120 from two divergent human immunodeficiency virus type 1 (HIV-1) isolates resulted in the development of seven hybridomas secreting monoclonal antibodies able to recognize regions of gp120 which are common among divergent strains of HIV-1. These monoclonal antibodies cross-reacted with env glycoproteins from one African (Rutz), one Haitian (RF), and three North American viral isolates, namely IIIB, MN, and 451 by either immunoblot or radioimmunoprecipitation assays. All recognized denatured gp120 in immunoblots with the exception of one which required a conformationally intact glycoprotein for reactivity. The gp120 epitopes identified by these antibodies were mapped by screening of an env gene library in the lambda gt11 expression system. Three out of four epitopes were found to reside in the amino-terminal half of gp120 (Cys9 to Cys35, Thr44 to Glu72 and Val108 to Met130), the other was located in the middle region (Thr221 to Ser255). By virtue of their extent of cross-reactivity these reagents might provide a unique resource for the detection of new viral isolates related to HIV-1.

Amino Acid Sequence↗
PubMed1992

Anti-hCG vaccines are in clinical trials.

Two vaccines inducing antibodies against human chorionic gonadotropin (hCG) have completed Phase I clinical trials, indicating the reversibility and safety of these vaccines. One is currently in Phase II efficacy trials in women in three major centres in India. The available data suggest that the vaccine prevents pregnancy above antibody titres of 50 ng/ml hCG bioneutralization capacity.

Antibodies↗
PubMed1992

Characterization of a neutralizing monoclonal antibody to the external glycoprotein of HIV-1.

The major neutralizing epitope on the external glycoprotein of HIV-1 was studied with an envelope-specific monoclonal antibody and with a human serum positive for antibodies to HIV-1 proteins, both of which were able to neutralize virus infectivity. The monoclonal antibody reacted specifically with gp120 from HIV-1IIIB, and was shown to neutralize infection of CEM cells by cell-free virions, and inhibited the formation of syncytia normally observed when uninfected cells are cocultured with HIV-1-infected cells. Similar neutralization of viral infection and inhibition of syncytia formation was also demonstrated by the HIV-1-antibody-positive human serum. By examining a number of overlapping peptides from a region of HIV-1 gp120 known to contain a neutralizing epitope, this epitope was localized between amino acids 307 and 320 (V3 loop) in the external glycoprotein molecule. The monoclonal antibody did not interfere with the binding of gp120 to CD4, or with the subsequent step of CD4-induced shedding of gp120 from the viral envelope. However, it blocked the proteolytic cleavage of the V3 loop by thrombin, suggesting that the antibody may be inhibiting the interaction of the loop with other membrane-bound proteins.

Amino Acid Sequence↗
PubMed1992

The Indian langur: preliminary report of a new nonhuman primate host for visceral leishmaniasis.

Described are the susceptibility of the Indian langur (Presbytis entellus) to Leishmania donovani and the consequent haematological and serum biochemical changes. The host response to antileishmanial chemotherapy and the immunological profile were also examined. Each langur was inoculated intravenously with 1 x 10(8) amastigotes; a spleen biopsy carried out on day 35 post-infection (p.i.) revealed 10-13 L. donovani bodies per 500 cell nuclei, which reached a maximum of 130-195 at death (day 105-110 p.i.). The infected monkeys lost body weight, developed severe anaemia, lymphocytosis, hyperproteinaemia, hypergammaglobulinaemia, hypoalbuminaemia and an increase in the level of alkaline phosphatase and alanine aminotransferase (AAT). Treatment with sodium stibogluconate (60 mg Sb5+ per kg body weight intramuscularly for 10 days) reduced the number of spleen parasites (0-1 amastigotes per 500 cell nuclei) but after the therapy the parasites appeared in the skin, which had previously been free of infection. Relapse occurred on day 30 post-treatment (10-24 amastigotes per 500 cell nuclei) and the parasites were resistant to repeat intensive therapy (120 mg Sb5+ per kg per day x 30 days). The stibogluconate treatment caused a proportionate reduction in the haematological and biochemical parameters to normal values except for alkaline phosphatase and AAT, which remained elevated. The level of IgG antibodies, which rose during the infection, rapidly fell to the pretreatment value following the first therapeutic schedule and then increased a second time coinciding with relapse. Our findings suggest that langurs could serve as acceptable models for human visceral leishmaniasis.

Animals↗