Biomedical subjects

G S Gupta

Publications and source records attributed to G S Gupta.

At least 19 recordsLinked to original sources

PubMed1998

Role of GABA receptor complex in low dose lindane (HCH) induced neurotoxicity: neurobehavioural, neurochemical and electrophysiological studies.

Lindane is widely used as an insecticide and scabicide in mammals. High doses in chronic exposures caused hyperexcitability and convulsions and impaired motor activity involving GABA-ergic mechanism. To investigate the role of GABA/Benzodiazepine mechanism in the neurotoxicity of low doses of lindane, rats were administered 2, 3, or 5 mg/kg orally for 90 days and behavioural, electrophysiological, and neurochemical studies were conducted. The animals exposed to lindane exhibited increased geotaxis and decreased spontaneous drug-induced locomotor activity (which further potentiated by phenobarbitone and increased after leptazol). The EEG of the treated rats showed high voltage slow-wave activity (HVSA) patterns with occasional spindles (9-10 HZ-amplitude of 100 uv). A significant increase (p < 0.01) in GABA levels in cerebellum and significant increase in benzodiazepine receptors in cerebellar membrane measured by (3H)flunitrazepam binding were observed in the animals exposed to 3 and 5 mg lindane. The study suggests that low dose chronic exposure of lindane causes neurobehavioral, neurochemical, and electrophysiological effects involving GABA-ergic mechanism(s).

Animals↗

PubMed1997

Immune responses of chemically modified homologous LDH-C4 and their effect on fertility regulation in mice.

PROBLEM: Lactate dehydrogenase-C4 (LDH-C4) is an iso-, allo- and auto-antigen of mammalian sperm. In spite of being cell specific, LDH-C4 does not induce infertility in females of homologous species after immunization. The present study examines the application of homologous LDH-C4 after chemical modifications in the immunological infertility of female mice. METHODS: LDH-C4 from testes of LACA mice was chemically modified by interacting it with gossypol (gossy-LDH-C4) and glucosylation with lactose (Glu-LDH-C4) in vitro and evaluated for immune responses and induced immunological infertility in allogeneic Balb/c mice after inoculation through intrarectal route using A1(OH)3 as adjuvant. RESULTS: Native LDH-C4, which elicited high antibody response after immunization with a primary (50 microgram) and three secondary doses (30 micrograms each) at an interval of 15 days each, did not reduce fertility significantly in mice. In contrast, study provides evidence that chemically modified LDH-C4 induces high infertility, since 85-100% of mice failed to conceive in two independent sets of experiments. Mice inoculated with modified LDH-C4 were associated with 2-3 fold anti-LDH-C4 antibody titre compared to antibody response elicited by native LDH-C4. Splenocytes from immunized non-pregnant mice were evaluated for polyclonal activation using Con A as mitogen. It was found that splenocytes primed with native LDH-C4 were significantly more stimulated than the non-immune control cultures. However cells primed with gossy-LDH-C4 were non-reactive to Con A and cells primed with glu-LDH-C4 were suppressed for ConA proliferation. CONCLUSIONS: It is concluded that LDH-C4-gossypol adduct offers a potential application in the induction of infertility of homologous species in marked contrast to native LDH-C4. Application of A1(OH)3 as adjuvant in the induction of immune response through intra-rectal route has been suggested.

Adjuvants, Immunologic↗

PubMed1997

LDH-C4-substrate binary complexes studied by intrinsic fluorescence method.

Lactate dehydrogenase-C4 (LDH-C4) has been studied in presence of substrates using intrinsic fluorescence measurements. Excitation maximum of LDH-C4 occurred at 282 nm whereas fluorescence emission maximum was obtained at 340 nm. Fluorescence intensities at 340 nm showed that ligands viz. NAD+, NADH, pyruvate and lactate quench the relative fluorescence intensities of LDH-C4 in a concentration dependent manner. NAD+ and NADH produced a maximum quenching between 92-93% while pyruvate and lactate quenched the fluorescence of LDH up to 29% and 21% respectively. Association constants (Ka) based on fluorescence measurements were 6.05 x 10(4)M-1, 20 x 10(4)M-1, 0.113 x 10(4)M-1 and 0.3 x 10(4)M-1, for NAD+, NADH, lactate and pyruvate respectively. Stern-Volmer constants (Ksv) show that NAD+ and NADH have single Ksv of 4.07 x 10(4)M-1 and 1.47 x 10(5)M-1, whereas lactate and pyruvate indicated quenching reaction to be made up of two components. Ksv at low and high concentration of lactate respectively were 0.645 x 10(2)M-1 and 0.05 x 10(2)M-1, whereas corresponding Ksv with pyruvate were 1.008 x 10(3)M-1 and 0.408 x 10(3)M-1. Low Ksv at higher concentrations suggested that the aromatic chromophores are located within a hydrophobic environment. Red shift in fluorescence maximum (lambda max) by 2nm with lactate and 6nm with pyruvate showed that interaction of these ligands with LDH-C4 exposes some buried chromophores of the enzyme to the surface.

Animals↗

PubMed1997

Quenching of intrinsic fluorescence of sperm specific LDH by optical isomers of gossypol.

Intrinsic fluorescence of LDH-C4 has been studied in the presence of optical isomers of gossypol. The study showed that fluorescence due to tryptophan residues after excitation of LDH at 282 nm is quenched by each gossypol enantiomere in a concentration dependent manner. Half of the maximum quench (Q50%) of enzyme occurred with gossypol (-) at 0.9 x 10(-4) mol/l and with gossypol (+) at 1.4 x 10(-4) mol/l showing a maximum quench (Qmax) of 45% and 65% respectively, with a corresponding association constant (Ka) of 1.0 x 10(4) l/mol and 0.4 x 10(4) l/mol. Stern-Volmer constant (Ksv) inferred that quenching of LDH comprises at least two components with two different Ksv values. Ksv(I) and Ksv(II) between LDH-C4 and gossypol (-) were 1.97 x 10(3) l/mol and 1.22 x 10(3) l/mol, and those between LDH-C4 and gossypol(+) were 2.3 x 10(3) l/mol and 1.56 x 10(3) l/mol. Smaller Ksv at higher concentrations of gossypol indicated that some of the tryptophan residues in LDH-C4 are deeply buried within a hydrophobic environment. There was no blue or red shift of LDH-C4 when interacting with either of the gossypol enantiomeres.

Animals↗

PubMed1996

Sex dependent immune responses by allogenic LDH isozymes.

Regulatory effects on polyclonal activation of primed splenocytes have been studied following immunization through the intrarectal route with allogenic sperm specific lactate dehydrogenase (LDH-C4) and somatic LDH from kidney. Results indicate that LDH primed cell proliferation by mitogens is dependent on the nature of the isozyme and sex of donor cells. Compared to somatic LDH, LDH-C4 was immunosuppressive for T cell proliferation in vitro and the effect was more significant with female splenocytes as compared to male spleen cells. However, the suppressive effect of LDH-C4, on B cell function was identical in both males and females. In contrast to the somatic LDH which did not produce alloantibody in significant amount, LDH-C4 was highly immunogenic in production of humoral antibody in female mice. Alloantibody formation in dams was substantiated with a similar degree of immune regulation of B cell functions as shown by lipopolysaccharide stimulation. The role of LDH-C4 in protection of allogenic sperm in the female genital tract has been suggested. However, it is concluded that recipients of sperm constituents through the intrarectal route are at greater risk for immune suppression and bacterial/viral infection.

Animals↗

PubMed1995

Renal toxicity after oral administration of lead acetate during pre- and post-implantation periods: effects on trace metal composition, metallo-enzymes and glutathione.

The present study was undertaken to compare the effects of 10-50 mg/kg b.wt. of Pb acetate after chronic treatment through oral gavage on: (a) the distribution of trace elements such as Fe, Cu, Zn, and Mn, (b) enzyme activity of delta-amino levulinic acid dehydratase (delta-ALAD) and alkaline phosphatase, and (c) glutathione (GSH) in kidney and (d) delta-ALAD in blood of pregnant and non-pregnant mice. Treatment with Pb acetate was given on every alternate day for 4 weeks prior to mating and for 3-4 weeks until pregnancy became apparent and confirmed by laporatomy. Lead administration reduced the rate of reproduction as assessed by number of living viable embryos. During normal pregnancy renal Cu, Fe and GSH tended to decline although non-significantly and continued to do so after lead administration. Mn was considerably and significantly elevated, whereas activity of delta-ALAD (non-activated) was quite low in pregnant mice. Following administration of Pb acetate, kidneys of pregnant and non-pregnant dams accumulated Pb in a dose-dependent manner, but as compared to non-pregnant mice, Pb increase in pregnant dams was significantly lower. Pb toxicity was associated with the loss of delta-ALAD in blood and kidney, but unlike the non-activated form of delta-ALAD, the dithiothreitol-activated form of delta-ALAD declined in a significant amount. The residual activity showed a high degree of negative correlation with endogenous Pb as well as with Zn/Pb ratio. Pb toxicity did not modify renal Fe, Cu, and Zn in the pregnant state, but reduced renal Fe during pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

PubMed1995

Molecular inactivation of testicular hyaluronidase in solid state after proton irradiation: a study based on target size, substrate binding and thermodynamic analysis of heat denaturation.

Dose response activity curve of testicular hyaluronidase (HDase) following proton irradiation in dry state follows complicated mechanisms which may involve multiple hits and multiple targets of variable sizes giving a constant G value of 1.66. Target analysis appears to be modified by slow recovery of activity when irradiated enzyme is brought to aqueous phase. However, pattern of irradiation at a dose of 1 x 10(5) to 8 x 10(5) Gy reveals that though binding affinity of enzyme to the substrate (hyaluronic acid) increases as shown by declining Km from 500 mg/l to 300-70 mg/l, the reaction rate of catalysis by irradiated HDase is decreased due to decrease in reaction velocity (Vmax: 266 versus 76 units at 8 x 10(5) Gy). Activation analysis of heat denaturation of nonirradiated HDase suggested the involvement of 78 kcal/mole of energy of activation (E*a) which declined to 63-52 k cal/mole after irradiation at 1 x 10(5) to 8 x 10(5) Gy for residual enzyme. The corresponding change in entropy of activation (delta S*) increased from a control value of -291 eu to -236 eu at 8 x 10(5) Gy. From thermodynamic analysis in association with recovery in aqueous phase, it is concluded that HDase is inactivated due to dissipation of proton energy among weak forces including H bonds associated with secondary/tertiary structure of molecules.

Animals↗

PubMed1994

Circadian rhythms of renal phosphohydrolases analysed by Cosinor method.

Rhythmometric analysis of hydrolytic enzymes of mouse kidney has been performed on circadian time scale using the F test. Significant rhythms were detected in glucose-6-phosphatase (G6Pase), inorganic pyrophosphatase (InPPase) and alkaline phosphatase (AlPase) on protein and fresh weight basis. Acrophase (time for peak activity) of G6Pase, InPPase and AlPase per mg protein was at 9.9 degrees (1.0 hr), 88.5 degrees (6.0 hr) and at 342.3 degrees (20 hr) respectively. ATPase, which did not show significant rhythm (mean +/- SD = 4.51 +/- 0.30), had a peak value at 32.1 degrees (2.14 hr) with an amplitude of 0.31 units on protein basis. However, G6Pase and AlPase oscillated with high amplitudes (0.18 and 0.71) across the mean value (mesor) of 0.68 +/- 0.3 and 1.43 +/- 0.46 units respectively and with a phase shift of 5 hr. Since G6Pase is a multicomponent and multifunctional enzyme having several overlapping enzyme activities (viz. InPPase), coordinated events of G6Pase, InPPase and ATPase in the regulation of daily renal functions have been mapped in the intact animals, along a physiologic time scale.

Animals↗

PubMed1994

Autoimmune-like activity of sperm specific LDH: a pathophysiological and electron microscopic study of atrophied testis and epididymis.

Hyper-immunization of male mice with human LDH-C4 evoked autoimmune reactions illustrated by the loss of LDH activity, associated histopathological changes in testes and epididymis and induction of sterility in mice. This was substantiated by the altered morphology of sperm mitochondria and plasma membrane, and by reduced number of cytoplasmic droplets as observed by electron microscopy. However, the presence of lymphoblasts and other lymphoid cells in testes indicated that the testicular damage is accentuated by activated T lymphocytes. It is concluded that immunization with human LDH-C4 produces lesions in mouse testis and epididymis, which are similar to experimentally induced autoimmune orchitis.

Animals↗

PubMed1994

Circadian rhythmometric analysis of hepatic phosphohydrolases with special reference to glucose-6-phosphatase and inorganic pyrophosphatase.

Rhythmometric analysis of a group of phosphohydrolases in mouse liver has been performed along a single 24 hr time scale. The presence of the rhythm was conducted by F test. Statistically significant circadian rhythm was detected in glucose-6-phosphatase (G6Pase) and inorganic pyrophosphatase (InPPase) activity expressed on fresh weight and protein basis. Both G6Pase and InPPase oscillated with a high amplitude of 0.44 U and 1.15 U respectively across the mean value (mesor) of 0.40 +/- 0.42 U and 2.81 +/- 1.14 U per mg protein and with a phase shift of 80 degrees (5.34 hr) among them. On the other hand, alkaline phosphatase (AlPase) did not show any rhythm whereas adenosine triphosphatase (ATPase) showed rhythmic activity on protein basis and oscillated across mesor of 1.84 +/- 0.5 U with an amplitude of 0.52. Acrophase (time for peak activity/mg protein) of G6Pase, InPPase and ATPase was found at 194.2 degrees (13.34 hr), 114.1 degrees (8.0 hr) and at 306.1 degrees (20.4 hr) respectively. AlPase, though did not show significant rhythm, had peak value at 231.8 degrees. Since hepatic G6Pase is a multicomponent and multifunctional enzyme with several overlapping activities (viz. InPPase), coordinated action of G6Pase and InPPase in the regulation of hepatic cell functions has been suggested.

Animals↗

PubMed1994

Effects of immunization with sperm specific lactate dehydrogenase with & without muramyl dipeptide as adjuvant.

The effect of sperm specific lactate dehydrogenase-C4 (LDH-C4) alone or with muramyl dipeptide (MDP) and Freund's complete adjuvant (FCA) on immune responses and breeding capacity have been studied in isogeneic C57 BI/Ks (H-2d) mice. Results per se suggested that LDH-C4 generates isoantibodies even in absence of adjuvant. Though MDP could be substituted for FCA as adjuvant, amplification by MDP of the antibody levels is reduced to half that obtained with FCA. LDH-B4 from kidney did not produce any antibody response under similar conditions. Systemic immunization with LDH-C4 did not reduce the overall pregnancy rate but reduced the frequency of embryo resorption and increased litter size significantly. However, mothers with foetal non-resorption had in general, lower antibody titres than the mothers showing foetal resorptions. The popliteal lymph node (PLN) assay for local graft versus host (LGVH) reaction revealed that the maternal lymphocyte cell competence was depressed by the sperm specific isozyme during gestation; depression in stimulation index was associated with the low antibody titre group. MDP, like LDH-C4 alone did not modify LGVH reaction significantly although it exerted a similar effect as LDH-C4 in embryo protection.

Acetylmuramyl-Alanyl-Isoglutamine↗

PubMed1994

Regulation of fertility in female mice after immunization with human sperm specific LDH: role in conception and contraception.

Effects on fertility of female mice after hyper-immunization with human lactate dehydrogenase-C4 have been studied at (50 + 25 x 4) and (50 + 50 x 4) micrograms doses of protein for two consecutive cycles of pregnancy. Results of heterologous immunization are described in relation to the role of LDH-C4 in conception and contraception. (i) Low dose of Immunization produced higher antibody (Ab) titer as tested before mating followed by higher infertility (70%) than by a higher dose of immunogen (46%), taking pregnancy as the end point. (ii) However, the results were reversed after second mating. For example, low dose of immunogen produced insignificant infertility, whereas high dose of immunogen gave 80% infertility. Similar order was shown by humoral Ab titer before mating i.e., high infertility was associated with high Ab titer and viceversa. (iii) It shows that the LDH-C4 induced infertility is reversible as is evidenced by low dose of immunization. (iv) In contrary to infertile dams, immunized fertile dams delivered a significantly higher litter size as compared to non-immunized control dams. It is concluded that immunity due to LDH-C4 is beneficial for embryo survival suggesting an immuno-suppressive nature of LDH-C4. Nonetheless, infertility ensues when immune-suppression is overcome by its own immune-activation.

Animals↗

PubMed1993

Modulation of allo-immune responses in vivo and in vitro by sperm specific lactate dehydrogenase-C4.

The role of sperm specific lactate dehydrogenase-C4 (LDH-C4) in allo-immune responses using mixed lymphocyte cultures (MLC) and cytotoxic T cell (CTL) generation in vitro and local graft versus host (LGVH) reaction and allograft enhancement in vivo has been ascertained. LDH was purified from testes (LDH-C4) and kidney (LDH-B4) of C57Bl/Ks mice. MLC and CTL were performed using C57Bl/Ks-anti A/J lymphocytes in presence of 10(-3)-1 micrograms LDH-B4 or LDH-C4 per culture. The MLC and CTL responses showed biphasic action depending on the dose of LDH-C4. Early MLC culture gave significantly low stimulation index at 10(-2)-10(-1) micrograms LDH-C4 as compared to non-treated control cultures. However, the MLC response in presence of LDH-C4 was not different from the LDH-B4 treated one which showed a similar biphasic trend. On the other hand, 51Cr release from YAC-222 target cells was practically abolished by LDH-C4 at 10(-3)-1(-1) micrograms, and this was strikingly different from LDH-B4 or non-treated cultures. LGVH reactivity as performed by using C57Bl/Ks lymphocytes along with LDH-C4 in (C57Bl/Ks x A/J) F1 hybrids indicated a suppression of stimulation index in primary and secondary (i.e. preimmunized in presence of LDH-C4 or LDH-B4) LGVH. Allograft enhancement of Sa I (A/J) in C57Bl/Ks mice in presence of LDH-C4, was delayed slightly but significantly during primary or secondary transplantation reaction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Modulation of biochemical and cytological profile of bronchoalveolar lavage constituents in rats following split-dose multiple inhalation exposure to methyl isocyanate.

1. Studies were carried out to explore the acute pulmonary effects of equal, split-dose, multiple inhalation exposures of rats to methyl isocyanate (MIC), (0.32 mg l-1, 8 min x 10 exposures) as reflected by alterations in bronchoalveolar lavage fluid (BALF) constituents and to evaluate recovery, if any, following survival in a MIC-free environment, 10 d after the last MIC exposure. 2. In the BALF of MIC-exposed rats, there was an increase in the total number of cells and the number of cells showing enhanced dye uptake and reduction of nitroblue tetrazolium chloride. The cell-free BALF showed increases in total protein, sialic acids and lactic acid contents and lactate dehydrogenase activity. 3. In rats exposed to MIC and sacrificed 10 d after survival in a MIC-free environment, there was a reduction in the cellular and biochemical constituents of BALF. The phagocytic potential of macrophages was, however, also decreased under this regime.

Administration, Inhalation↗

PubMed1993

Expression and regulation of insulin-like growth factor-I in the rat incisor.

Growth factors play an important role in the regulation of cell growth, division and differentiation. In this study the distribution and regulation of insulin-like growth factor-I (IGF-I) in the continuously erupting rat incisor was determined by immunohistochemistry. Results were evaluated both visually and with a computer-based image analysis system. The distribution and intensity of IGF-I immunoreactivity varied with developmental stage of the rat incisor. Strong IGF-I immunoreactivity was observed in differentiating odontoblasts and ameloblasts. The most intense immunoreactivity was observed in secretory ameloblasts, secretory odontoblasts and in maturation ameloblasts. Staining was weak or absent in post-secretory ameloblasts but persisted in post-secretory odontoblasts. Weak to moderate immunoreactivity was also seen in cells of the stratum intermedium and in the reduced enamel epithelium. Surrounding alveolar bone showed strong IGF-I immunoreactivity in osteoblasts and in the stratum basale and stratum spinosum of the adjacent labial gingival epithelium. In order to assess the role of GH in IGF-I expression, GH (65 micrograms/100 g bw) was administered for six days to dwarf GH deficient rats, producing a significant increase in body weight (P < 0.01). Measurements at different stages of odontogenesis showed that the staining intensity of secretory ameloblasts (P < 0.01) and maturation ameloblasts (P < 0.001) was significantly different between untreated and treated animals. These results indicate that IGF-I is present in cell populations of the enamel organ of the rat incisor found previously to exhibit growth hormone receptors, and that expression of IGF is GH dependent.(ABSTRACT TRUNCATED AT 250 WORDS)

Ameloblasts↗

PubMed1993

Histomorphological changes in lung of rats following exposure to wood smoke.

Rats were exposed to repeated, intermittent exposure to smoke generated from combustion of 1g wood/15 min, total period for 75 min daily under dynamic exposure conditions, over a period of 15, 30 and 45 days. First 15 days exposure caused mild bronchiolitis, hyperplasia and hypertrophy of bronchiolar epithelial lining cells, some necrosed lining cells desquamated into lumens, congestion of parenchymatous blood vessels, oedema, hyperplasia of lymphoid follicles, peribronchiolar and perivascular infiltration of polymorphonuclear cells, and mild emphysema. These lesions progressed further during 30 and 45 days of exposure, though emphysematous changes remain constant. By 30 days and 45 days, hyperplastic and hypertrophic changes of bronchioles become quite marked, with mononuclear cells infiltration and alveolar septa thickening. Hematological studies show marginal alterations in hemoglobin levels, ESR, PCV and TLCS during 15 days, where as significant changes in eosinophil were observed during 30 and 45 days, and ESR during 45 days only. The results indicate progressive pathomorphological pulmonary lesions with subsequent exposure to wood smoke in controlled conditions.

Animals↗

PubMed1992

Toxic effects of omega chrome red ME and its treatment by adsorption.

Toxic effects of Omega Chrome Red ME, a popular textile dye, on the nitrogen fixing cyanobacterium Nostoc calcicola were studied. The growth of N. calcicola was found to be suppressed at 10 and 20 mg liter-1 initial concentrations of dye, whereas a low initial concentration of 5 mg liter-1 slightly favors growth. Removal of the dye was carried out by adsorption using some cheap and unconventional adsorbents like coal, fly ash, wollastonite, and china clay. It has been observed that, in all cases, the low adsorbate concentration, the low temperature, and an acidic medium favor the dye removal process. The process of uptake follows first-order adsorption rate expression and obeys Langmuir's model of adsorption. The removal process is also partially diffusion controlled. Thermodynamic and pH studies were run to explain the results.

Adsorption↗

PubMed1992

Ranking toxicity of industrial dusts by bronchoalveolar lavage fluid analysis.

Female wistar rats were inoculated intratracheally with 10 mg/ml suspensions of various dusts, viz: quartz, fly ash, mica and corundum in physiological saline. Biochemical markers of bronchoalveolar lavage fluid (BALF) were analysed 8 days after the instillation of the dusts. Elevated levels of proteins, sialic acid and phospholipid contents and the activity of lactate dehydrogenase correlated well with the degree of the known fibrogenic potential of different dusts in the lungs in the following order, quartz greater than fly ash greater than mica greater than corundum. beta-glucuronidase activity, was however, only elevated in the quartz inoculated group of rats. It is suggested that biochemical constituents of BALF analysed shortly after the exposure to different dusts can be useful to mirror alterations in the tissue response to mineral dusts.

Aluminum Oxide↗