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Biomedical subjects

B N Joshi

Publications and source records attributed to B N Joshi.

At least 19 recordsLinked to original sources
PubMed2000

Melatonin counteracts the stimulatory effects of blinding or exposure to red light on reproduction in the skipper frog Rana cyanophlyctis.

An earlier study reported that blinding or exposure to red light stimulates reproduction in R. cyanophlyctis. The present study investigates whether melatonin counteracted blinding- or red-light-induced ovarian stimulation. The study consisted of two experiments carried out during the prebreeding (March/April) and breeding phase (August/September) of the reproductive cycle of this species. In experiment 1, exposure to red light for 30 days increased (P<0.01) the gonadosomatic index (GSI) and oviductal weights, whereas exposure to continuous darkness for the same duration decreased (P<0.01) the GSI. Data on follicular kinetics revealed an increase in vitellogenic oocytes and decrease in previtellogenic oocyte numbers following exposure to red lights. Exposure to continuous dark on the other hand increased previtellogenic oocyte and decreased vitellogenic oocyte numbers. In experiment 2, the GSI and the number of vitellogenic oocytes increased in blinded and red-light-exposed frogs. However, treatment with melatonin (15 microg sc) for 30 days to both these groups of frogs reduced the GSI, oviductal weights, and the number of vitellogenic oocytes. The results both confirm earlier finding that blinding and exposure to red light stimulate ovarian growth and also demonstrate that melatonin counteracts blinding or red-light-induced stimulation of ovarian activity. This apart, the findings from experiment 1 demonstrate for the first time that exposure to continuous dark inhibits ovarian growth in this species. Ocular melatonin seems to play an important role in regulation of reproduction in this species.

Animals↗
PubMed1999

Pearl millet cysteine protease inhibitor. Evidence for the presence of two distinct sites responsible for anti-fungal and anti-feedent activities.

Recently, pearl millet cysteine protease inhibitor (CPI) was, for the first time, shown to possess anti-fungal activity in addition to its anti-feedent (protease inhibitory) activity [Joshi, B.N. et al. (1998) Biochem. Biophys. Res. Commun. 246, 382-387]. Characterization of CPI revealed that it has a reversible mode of action for protease inhibition. The CD spectrum exhibited a 35% alpha helix and 65% random coil structure. The intrinsic fluorescence spectrum was typical of a protein devoid of tryptophan residues. Demetallation of Zn2+ resulted in a substantial change in the secondary and tertiary structure of CPI accompanied by the complete loss of anti-fungal and inhibitory activity indicating that Zn2+ plays an important role in maintaining both structural integrity and biological function. The differential response of anti-fungal and inhibitory activities to specific modifiers showed that there are two different reactive sites associated with anti-fungal and anti-feedent activity in CPI located on a single protein as revealed from its N-terminal sequence data (AGVCYGVLGNNLP). Modification of cysteine, glutamic/aspartic acid or argnine resulted in abolition of the anti-fungal activity of CPI, whereas modification of arginine led to an enhancement of the inhibitory activity in solution. Modification of histidine resulted in a twofold increase in the protease inhibitory activity without affecting the anti-fungal activity, whereas modification of serine led to selective inhibition of the protease inhibitory activity. The differential nature of the two activities was further supported by differences in the temperature stabilities of the anti-fungal (60 degrees C) and inhibitory (40 degrees C) activities. Binding of papain to CPI did not abolish the anti-fungal activity of CPI, supporting the presence of two active sites on CPI. The differential behavior of CPI towards anti-fungal and anti-feedent activity cannot be attributed to changes in conformation, as assessed by their CD and fluorescence spectra. The interaction of CPI modified for arginine or histidine with papain resulted in an enhancement of CPI activity accompanied by a slight decrease in fluorescence intensity of 15-20% at 343 nm. In contrast, modification of serine resulted in inhibition of CPI activity with a concomitant increase of 20% in the fluorescence intensity when complexed by the enzyme. This implies the involvement of enzyme-based tryptophan in the formation of a biologically active enzyme-inhibitor complex. The presence of anti-fungal and anti-feedent activity on a single protein, as evidenced in pearl millet CPI, opens up a new possibility of raising a transgenic plant resistant to pathogens, as well as pests, by transfer of a single CPI gene.

Antifungal Agents↗
PubMed1999

Purification and characterization of a cell-surface lectin (Lectin II) from Agrobacterium radiobacter NCIM 2443.

A lectin was isolated from Agrobacterium radiobacter cell surface and purified. It is a monomer of 40 kDa as shown by SDS-PAGE. The lectin has a pI of 9.15 and amino acid composition of the lectin shows that 44% of the amino acids are hydrophobic. The lectin agglutinates rabbit erythrocytes but does not agglutinate human erythrocytes. It does not show specificity for monosaccharides except for D-glucosamine. Fetuin and its N-linked glycopeptide also inhibit the activity of the lectin but greater inhibition is shown by locust bean gum and Nicotiana tobaccum (tobacco) tissue extracts.

Amino Acids↗
PubMed1998

Cysteine protease inhibitor from pearl millet: a new class of antifungal protein.

A cysteine protease inhibitor exhibiting antifungal activity from pearl millet seeds has been purified to homogeneity by ammonium sulphate precipitation and chromatographic procedures involving CM- sephadex and SP-sepharose cation exchange columns. The molecular characterization has revealed its molecular mass as 24 kD and isoelectric point 9.8. The amino acid composition data shows presence of high content of serine and glycine (34 residues/mole) and absence of tryptophan. The inhibitor exhibits potent antifungal activity against Trichoderma reesei, a dead wood fungus with minimum inhibitory dose to inhibit mycelial growth or spore germination is as low as 1 microgram/ml (250 ng/disc). In addition to Trichoderma reesei, the antifungal activity is observed against some important phytopathogenic fungi, namely, Claviceps, Helminthosporium, Curvularia, Alternaria and Fusarium species. To the best of our knowledge, a cysteine protease inhibitor as an antifungal protein is reported for the first time from a plant system.

Amino Acids↗
PubMed1998

Changes in ovarian follicular kinetics in intact blinded and parietal shielded frogs exposed to different spectra of light.

Ovarian follicular kinetics and gravimetric changes in the ovary and oviduct were studied in intact, blinded (BL), and parietal shielded (PS) skipper frog Rana cyanophlyctis exposed to different light spectra. The gonadosomatic index (GSI) increased (P < 0.01) in intact and PS frogs, held in green, yellow, and red light. The maximum increase was in red light followed by yellow and green. The GSI of BL frogs also increased (P < 0.001) in white, green, yellow, and red light. However, the GSI of blinded and parietal shielded (BLPS) frogs increased (P < 0.001) in white and all the colored lights compared with controls. The percentage changes of oviductal weights were similar to the ovarian weights. Data on follicular kinetics revealed a decrease in previtellogenic oocytes in intact and PS frogs placed in blue, green, yellow, and red lights. The previtellogenic oocytes of BL and BLPS frogs decreased in white light, while their number did not vary significantly in other spectra. The vitellogenic oocytes of intact and PS frogs increased in all the spectra, with a maximum increase in red. The vitellogenic oocyte count increased in both BL and BLPS frogs held in white light. The green, yellow, and red spectra stimulated vitellogenic oocyte count of BL frogs. The response of BPLS frogs was similar except for a slightly decreased oocyte count in red light. The pattern of vitellogenic oocyte counts in general exhibited a negative correlation with previtellogenic oocyte counts in all the frogs. The atretic follicular numbers did vary significantly in all of the groups. Red light stimulated ovarian activity maximally, followed by yellow and green. As the ovary exhibited varied response to the different spectra of light even in BL frogs, it may be concluded that extraretinal perception of colored light occurs in this species.

Animals↗
PubMed1997

Melatonin and exposure to constant light/darkness affects ovarian follicular kinetics and estrous cycle in Indian desert gerbil Meriones hurrianae.

Melatonin mediates photoperiodic influence on reproduction and constant light and darkness affect pineal biosynthesis of melatonin. The present study was undertaken to assess the effects of melatonin and drastic photoperiodic changes on reproduction in a tropical desert species with a fossorial lifestyle. Ovarian follicular kinetics and estrous cycle were studied in the Indian desert gerbil Meriones hurrianae, after treatment with melatonin and exposure to constant light (LL) and darkness (DD) regimes. Melatonin treatment increased (P < 0.001) ovarian weights without changing the uterine weights. While exposure to LL decreased (P < 0.001) both ovarian and uterine weights, exposure to DD had no effect on these weights. Follicular kinetics of growing and regressing follicles revealed that ovaries of melatonin-treated and DD-exposed animals had significantly more growing follicles. Melatonin treatment increased all types of growing follicles, especially antral and Graafian follicles. Exposure to DD increased all types of growing follicles, with the medium sized antral and Graafian follicles being significant (P < 0.01). In contrast to stimulation of follicular growth by melatonin and DD, LL caused regression of all stages of follicular growth and also reduced the number of small preantral follicles. Melatonin treatment increased (P < 0.001) the length of estrous cycle (5.08 to 7.29 days). Gerbils treated with melatonin, exposed to LL and DD, had a longer (P < 0.001) metestrus. Animals held in LL, had the least number (P < 0.001) of estrous smears (1 in 30 days). The results suggest that melatonin is involved in growth of ovarian follicles in the Indian desert gerbil.

Animals↗
PubMed1997

Photoperiod and melatonin-induced changes in male reproduction in Indian desert gerbil, Meriones hurrianae (Jerdon).

Exposure to continuous darkness and chronic treatment with melatonin, for six weeks, stimulated reproduction in the male Indian desert gerbil as evidenced by morphometric data. Exposure to continuous light, for same duration, on the other hand inhibited reproduction. The results are opposite to those reported from similar studies on temperate zone species. Surface areas of abdominal scent glands increased following both, exposures to continuous darkness and the treatment with melatonin. Exposure to continuous light decreased the scent gland surface area. Assessment of scent gland activity could be useful in evaluation of reproductive function as they are dependent on sex steroids. Melatonin that mediates photoperiodic influence on reproduction is not always inhibitory to gonads.

Animals↗
PubMed1996

Ovarian follicular kinetics in response to enucleation and parietal shielding in Rana cyanophlyctis.

Ovarian follicular kinetics, gravimetric changes in the ovary and the oviduct, were studied in the skipper frog, Rana cyanophlyctis, following enucleation (ENX) and parietal shielding (PS), during the breeding reason. PS was to prevent extraoptic photoreception by the pineal via the frontal organ. The gonadosomatic index (GSI) was high in ENX and PS frogs (P < 0.01). The highest GSI occurred in frogs that were both ENX and PS (P < 0.01). Percentage of oviductal weight decreased (P < 0.01) in PS frogs and increased (P < 0.01) in those subjected to both ENX and PS. Oviductal weight of ENX frogs was unchanged. Data on follicular kinetics revealed a decrease (P < 0. 01) in the ENX group in the number of first growth phase (FGP) oocytes and atretic follicles (AF), whereas the number of medium-sized secondary growth phase (MSGP) oocytes and large-sized secondary growth phase (LSGP) oocytes increased (P < 0.01). There was a similar pattern of follicular kinetics in ENX+PS frogs. However, following PS, the number of FGP oocytes and AF increased (P < 0.01) and decreased (P < 0.01) in the MSGP and LSGP oocytes. The lower oviductal weight in PS frogs correlated with reduced SGP oocytes, the major source of estrogen. The importance of lateral eyes and the frontal organs in the transduction of environmental signals to the neuroendocrine-reproductive axis is indicated in this species. That recruitment of FGP to SGP is inhibited only in the PS group suggests that the frontal organ plays an important role in ovarian follicular kinetics.

Animals↗
PubMed1996

Influence of melatonin on blood morphology in male albino mice.

Influence of melatonin on diurnal changes in hematological profiles was examined in male albino mice, at six hourly interval. Melatonin treatment either once daily (25 micrograms at 1700 hrs) or twice daily (25 micrograms at 0900 and 1700 hrs) for two weeks resulted in an alternation in the RBC rhythm. Generally the effect was suppressive though at some times the counts were marginally increased. Total WBC counts were increased and there was apparently a change in their rhythm too. Interestingly the differential WBC counts exhibited different patterns in, once and twice daily melatonin-treated mice. Melatonin given once daily predominantly stimulated the absolute lymphocyte count whereas the twice daily regimen predominantly stimulated the neutrophil count. This perhaps is related to the varied exposures of the animals to melatonin in the two experimental setups. Erythrocyte indices like mean cell volume, mean cell hemolglobin and mean cell hemoglobin concentration in both the experiments correlated with hemoglobin and hematocrit values. It may be concluded that melatonin has a modulatory role in hemopoiesis and its rhythms. The stimulatory effect of melatonin on WBC supports its purported immunopotentiating action.

Analysis of Variance↗
PubMed1995

Effect of long-term administration of melatonin on eyes and harderian glands in albino rat.

Pineal hormone melatonin is also synthesized by the ocular structures, retina and Harderian gland. The physiological significance of such production is not fully understood. Daily treatment with melatonin (25 micrograms/rat) for 8 weeks led to an increase in weight of eye balls and fluid within it (P < 0.05). Weight of the Harderian gland in the control and treated rats was not significantly different, though the histological observation revealed an increase in cell height and reduction in lumen diameter in the treated animals. Inter luminal deposits of porphyrin content appeared to be more in the Harderian glands of melatonin treated rats. The retinal tissue indicated degenerative changes in the melatonin treated rats. The results suggest melatonin's influence on level of ocular fluid, structure/function of retina and harderian gland.

Animals↗
PubMed1995

Effect of long-term administration of melatonin on adrenal histomorphology of the Indian desert gerbil Meriones hurrianae Jerdon.

Melatonin treatment for 8 weeks resulted in a significant increase (P < 0.01) in weights of the adrenal glands, in both male and female, M. hurrianae. The weight increase was not related to the time of administration of melatonin. Histology of adrenal glands in melatonin treated gerbils revealed a stimulatory response. In treated gerbils, adrenal cortex exhibited significant hypertrophy with enlarged nuclei and degranulated cytoplasm. Signs of hypertrophy were also evident in the adrenal medulla. The physiological significance of this response is not yet clear. It remains to be clarified if the hypertrophy indicates enhanced synthesis of corticosteroids.

Adrenal Glands↗
PubMed1993

Studies on the migration of micronucleated erythrocytes from bone marrow to the peripheral blood in irradiated Swiss mice.

Micronucleated polychromatic (mn-PCE) and normochromatic erythrocytes (nm-NCE) were enumerated in the bone marrow and peripheral blood of Swiss male mice at different time intervals following whole-body (1.0 Gy) gamma-irradiation. Polychromatic cells migrated to the peripheral blood soon after their formation in the bone marrow and mn-PCE achieved a frequency close to that of the bone marrow with a delay of about 12 h. The optimal time for peripheral sampling was found to be about 36 h after radiation exposure. The frequency of mn-NCE in bone marrow and peripheral blood showed only a moderate and gradual increase till 60 h, and was much lower in the latter. In another experiment, mice irradiated with 0.42 Gy gamma-rays (0.21 Gy/h) once a day for 5, 10 or 15 days (5 days per week) showed a cumulative dose-dependent increase in the levels of mn-NCE in the peripheral blood, sampled at 7 or 21 days after the last exposure. These observations demonstrate persistence and accumulation of mn-PCE in the peripheral blood of mice during repeated exposure to ionizing radiation, and the sampling could be delayed up to several days after the last exposure. Thus, peripheral mn-PCE, scored between 24-48 h following irradiation, can be conveniently used to measure acute chromosomal damage induced by ionizing radiation in the bone marrow erythroblasts of mice, while peripheral mn-NCE are suited to monitor accumulated damage during chronic/repeated exposure.

Animals↗
PubMed1989

Salmonella newport isolations from a general hospital in Pune.

Salmonella newport had not been reported from the Pune (Maharashtra) area till 1983. After the first isolate in 1984, the isolation rate showed a significant increase. A total of 64 isolates todate (two isolates in 1984, six in 1985 and 56 in 1986) have been encountered at Sassoon General Hospital, Pune. It has been commonly isolated from adults (54.7 per cent). Most of the strains were multi-drug resistant.

Adolescent↗
PubMed1988

Effects of aggressive encounters on pineal melatonin formation in male gerbils (Meriones unguiculatus, Cricetidae).

Pineal N-acetyl-transferase activity and radioimmunoassayable melatonin levels were determined in adult male gerbils subjected to aggressive encounters using the intruder-model. In the first experiment, a single encounter of 3 min was applied in the afternoon to intact and to animals with sympathetically denervated pineal organs. Compared with controls, both stressed groups demonstrated a drastic decrease in N-acetyl-transferase activity followed by a slow recovery. In both groups there also occurred a marked change in pineal melatonin content: in intact animals pineal melatonin levels were elevated immediately after the encounter; thereafter, melatonin values decreased. In animals bearing denervated pineal organs melatonin levels fell as a consequence of the encounter. In a second experiment, intact gerbils experienced four daily encounters of 1 min for one week. Thereafter the nocturnal formation of melatonin was studied. In comparison with untreated controls, the repeatedly stressed animals demonstrated a temporal delay in the rise of both N-acetyl-transferase activity and melatonin. Since the pineal organ is able to transduce events of the social environment into an endocrine message--as set forth by both our experiments--the pineal organ might play an important role within central processing of social stress.

Acetyltransferases↗