Histophysiological responses in ovary and liver of Cyprinus carpio after short term exposure to safe concentration of mercuric chloride and recovery pattern.

Effect of short term (7 days) exposure to safe concentration of HgCl2 (0.5 ppm) and changes 7 days after withdrawl of the treatment on his ophysiologyof ovary and liver in yearlings of Cyprinus carpio were assessed during active phase of reproductive cycle. Noticeable degenerative histophysiological changes were observed in both ovary and liver after exposure which were more prominent in the group with abnormal behaviour. After withdrawl of the HgCl2 treatment the recovery was apparent in both organs but was more appreciable in liver. These observations indicated that even safe concentration of HgCl2 might not be fully devoid of deleterious influence on reproductive functions in Cyprinus carpio.

Ovarian development in Labeo dyocheilus (McClelland) during active reproductive phase under captive and wild conditions.

Ovarian development in Labeo dyocheilus was assessed during active reproductive phase under ambient environmental conditions in captivity and wild. Increasing day length and water temperature seemed favourable for ovarian development in female L. dyocheilus under both conditions. Gonadosomatic index (GSI) was lower in May and higher in July in captivity (6.168 and 13.366) and wild (5.798 and 16.166) respectively Ovarian development started little bit in advance in captivity with late yolk vesicle stage oocyte in May when yolk globule stage oocytes were prominent in wild. Fully grown oocytes with germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) stages were observed in July in both conditions. Degree of transformation of developing oogonials into advance stage oocytes was observed to be better in wild fish compared to captivity reared ones. The histophysiological changes in liver corresponded well to the progression of ovarian development. Successive granulation and vacuolization of hepatocyte cytoplasm were indicative of augmented synthetic activity and probably mobilization of energy content for oocyte growth. These observations indicated that normal ovarian development of L. dyocheilus under captivity in Tarai region of Uttarakhand would be useful for success of its seed production in captivity for stock augmentation in wild or species diversification in aquaculture.

Behavioural and hematological responses of Cyprinus carpio exposed to mercurial chloride.

The static bioassay experiments were conducted to study the hematological and behavioural responses of Cyprinus carpio after exposure to mercuric chloride. Three different concentrations i.e. 0.5, 1.0 and 1.5 ppm of mercuric chloride (HgCl2) were used for 8, 16, 24, 48, 72, 96 and 124 hrs for evaluating behavioural responses. Hematological responses were assessed after 60 days exposure to 0.1 ppm of HgCl2. The changes in body colour, movement, sluggishness or activeness, disbalance etc. constituted the observations on behavioural responses. Hematological parameters included total erythrocyte (TEC) and leukocyte (TLC) counts and hemoglobin (Hb) levels. Body colour had changed in all groups after 48 hours. The fish exposed to 0.5 ppm HgCl2 did not show any abnormal activity except colour change throughout the experiment. Though no sign of distress was observed initially in groups exposed to 1.0 and 1.5 ppm HgCl2 but abnormal posturing, disbalance and sluggishness became apparent after 72 hrs and all specimens of 1.5 ppm group had died within 124 hours. TEC and Hb levels decreased whereas TLC increased in both male and female specimens of C. carpio after exposure to 0.1 ppm HgCl2 for 60 days. These observations indicated that mercurial toxicity even at low levels, caused adverse effects on body colour, behavioural responses and hematological parameters like TEC, TLC and hemoglobin levels in C. carpio.

Seasonal ovarian cycle in freshwater teleost, Labeo rohita (Ham.) in Tarai region of Uttaranchal.

Seasonal ovarian cycle, based on histophysiological changes, and correlative variations in liver were studied in an Indian major carp, Labeo rohita reared in captivity under ambient environmental conditions in Tarai region of Uttaranchal. The oocyte features used for marking ovarian cycle included nuclear-ooplasmic ratio, yolk vesicle incorporation into ooplasm, number and size of nucleolus and differentiation of follicular layer. Based on these features the ovarian cycle in L. rohita was divided into five phases i.e. resting, preparatory, pre-spawning, spawning and post-spawning/regression. The resting and preparatory phases were characterized by the dominating presence of chromatin-nucleolar and yolk-vesicle stage oocytes. The pre-spawning phase ovary was marked by the prevalence of yolk globule stage oocytes having cortical alveoli and differentiating follicular wall. The oocytes containing densely packed yolk-granules and large masses of yolk vesicles and also exhibiting germinal vesicle movement (GVM) signified the ovary of spawning phase. The ovary in regression phase contained pre-ovulatory atretic oocytes and significant number of oogonia. The histophysiological changes in liver showing augmented biosynthetic activity during preparatory and pre-spawning phases seemed well correlated with the gonadal development. The increasing water temperature and day-length apparently was favourable for the gonadal development in L. rohita.

Fibroblast growth factor in chick osteogenesis.

In vivo effects of FGF on osteogenesis in the chick embryo were evaluated. Day-11 embryos were injected with FGF followed by radiotracers. 3H-thymidine labelling of osteogenic cells was significantly higher following FGF administration; 3H-proline labelling over bone matrix was greater in the controls. Cartilage cells and matrix were sparsely labelled indicating a low level of metabolic activity. These data provide the first in vivo evidence that FGF stimulates osteogenic cell proliferation, while inhibiting production of bone matrix collagen. A role for FGF in embryonic osteogenesis and in fracture repair is suggested.

An autoradiographic study of the uptake of tritiated proline by osteoblasts during hibernation.

Twenty-four LSH and LVG strain golden hamsters, Mesocricetus auratus, were used. Experimental animals were maintained at 5 C and allowed to hibernate. Control animals were kept at 27 C. Six animals (3 experimental, 3 control) were injected subcutaneously with 1 microCi of 3H-proline/gm body wt. (Spec. act. 3 Ci/mM) after hibernation lasting 12 hours, 1 day, 3 days, or 7 days. Animals were killed 1 hour after injection and autoradiographs were prepared from 5 microns thick decalcified sections of femurs. A greater number of endosteal cells were labeled than periosteal cells and also exhibited a greater magnitude of labeling throughout the study. Differences between endosteal and periosteal cells both in percentage of cells labeled and magnitude of labeling were maximum in control animals and progressively decreased with increasing periods of hibernation. A reduction in synthesis of matrix proteins during the early period of hibernation was seen and was attributed to a significant reduction both in average cell activity and in the number of active cells during hibernation. The latter phenomenon apparently made a large contribution to the reduced matrical synthesis. 3H-proline uptake by osteoblasts probably reflects the reduced requirements of matrical synthesis during hibernation.

Autoradiographic investigation of circadian rhythms in alveolar bone periosteum and cementum in young mice.

This report presents circadian rhythms in cell proliferation of alveolar bone periosteum and cementum of the maxillary first molars of male 5-week-old BNL, Swiss albino mice which were maintained on a 12 hr light/dark cycle. Mice were injected with 3H-TDR (luCi/gm. body wt.) 1 hr prior to sacrifice and killed every 3 hrs for 24 hrs starting at 9 a.m. Maxillae were decalcified, routinely processed histologically and autoradiographs prepared. Cell labeling indices of alveolar bone and cementum mesial to the first molar were determined. Alveolar bone periosteal and cemental cells show circadian rhythm in their DNA synthetic processes. Peaks in percent labeling exhibit higher values than previously reported for nontraumatized, normal dental periosteum and cementum. While the outer periosteum reveals a single 24 hr peak (6 p.m.), inner periosteum and cementum reveal two ultradian peaks 9 to 12 hrs apart involving both light and dark periods. Rodents are nocturnal, but high peaks are also evident in the light periods, consequently, not all peaks are synonymous with the period of animal activity and feeding. Although the single daylight peak of the outer periosteum may indicate growth of that surface at night to about noon, the double peaks exhibited by inner periosteum and cementum indicate light/dark, continuously active surfaces in terms of DNA synthesis and growth.

A comparative study of the polypeptides of mammalian peripheral nerve myelin.

Polypeptide components of peripheral nerve myelin prepared from sciatic nerves of mouse, rat, rabbit, cat, guinea pig, monkey, dog and chicken were analyzed using a 20% polyacrylamide slab-gel electrophoresis system. Using this technique, we have demonstrated that there are at least two polypeptides present in the region of P1 protein (mol. wt. 18,000 and 16,000), and two closely moving components in the region of P2 protein (mol. wt. 13,500). A polypeptide (mol. wt. 10,500) is present in significant amounts only in mouse sciatic nerve myelin. Staining with periodic acid-Schiff reagent shows that in addition to P0 protein (mol. wt. 28,000), the major glycoprotein of peripheral nerve myelin, there are several minor glycoprotein components in both the high and low molecular weight regions.

The effects of cold-stress. Hibernation, and prolonged inactivity on bone dynamics in the golden hamster, Mesocricetus auratus.

The effects of cold-stress and hibernation on bone dynamics in the femurs of hamsters were investigated using histometric analyses. Control animals were maintained at 27 degrees C for 90 days; experimental animals were kept at 5 degrees C and hibernated for 7, 15, 21, 50, or 90 days. Histometric analyses of cross sections indicated that bone diameter and cortical thickness at the femoral midshaft increased after 83 days of extreme cold and 7 days of hibernation but decreased significantly after 69 days of cold stress and 21 days of hibernation. Osteoporosis was evident although the number of osteons per unit area of bone increased during hibernation. An initial decrease in the number of non-Haversian longitudinal vessels per unit area of bone was seen in experimental animals which was apparently related to a corresponding reduction in cortical thickness. Lacunar area increased in these animals, suggesting that osteocytic osteolysis may be a significant mechanism for calcium regulation during hibernation.

Circadian rhythms of gingival epithelium in young mice.

Five-week-old Brookhaven National Laboratory (BNL), Swiss-albino male mice were maintained on a 12-hour light/dark cycle and were subcutaneously injected with 3H-thymidine (1 micro Ci/gm body weight) one hour prior to sacrifice. Twenty-four mice (three each time-period) were killed at 3 hour intervals for 24 hours. Autoradiographs were prepared from 5-micron thick paraffin embedded decalcified sections of maxillary first molars and surrounding tissues. Labeling index was determined over the cells of the basal layer of the gingival epithelium. Crevicular and crestal gingival epithelium revealed a circadian rhythmic pattern in DNA synthesis in which two peaks were evident. Peak labeling of 5-week-old mouse gingiva exhibited a higher value than previously reported for nontraumatized gingiva. Rodents are nocturnal, as was peak activity, but a second peak is also evident in the light period. Consequently, not all peaks are synonymous with the period of animal activity and feeding. The observed continuous high labeling indices represent a significant potential of gingival epithelium to replenish its cells in response to physiological wear and tear.

Autoradiography of [3H]-proline uptake by fibroblasts of the periodontal ligament and odontoblasts of the rat incisor after guanethidine-induced sympathectomy.

The uptake of [3H]-proline was monitored by quantitative autoradiography at 30, 45 and 90 days of age. Grain counts in sympathectomized rats did not vary significantly from control values at any time. Thus the sympathetic nervous system does not influence the metabolic activity of these cells, which agrees with morphologic evidence that the innervation of the rat incisor may not be autonomic.

Autoradiographic evaluation of circadian periodicity in cell proliferation in the periodontal ligament of the young mouse.

Five-week-old BNL male mice were maintained on a 12-h light/dark cycle, subcutaneously injected with [3H]-thymidine 1 h before death and killed every 3 h for 24 h. Autoradiographs were made from demineralized, paraffin-wax embedded sagittal sections of maxillae. The labelling index was calculated for fibroblast-like cells of the periodontal ligament mesial to the first molar. A bimodal distribution of labelled cells was seen. Labelling index was highest at 0900 h (light period); a second, lower, peak was seen at 2400 h (dark period). Cell proliferation was significantly increased during the light period. The increased activity during the dark period may represent a superimposed secondary, ultradian, rhythm, differences in periodontal cell subtypes or may be related to the feeding and activity cycle of the mouse. Thus there is a 24-h cyclic rhythmicity in the proliferation of periodontal-ligament cells in the mouse.

The early effects of surgical sympathectomy on bone resorption in the rat incisor socket.

Earlier studies have shown that one week after sympathectomy in the rat, both bone apposition and resorption are significantly altered. Resorption in the incisor socket was now examined 1, 3 and 5 days after surgical sympathectomy. Morphometric analyses indicated that the total resorbing surface, active resorbing surface and number of osteoclasts per socket were increased one day after sympathectomy; the area and length of the bone surface of the socket were not significantly altered. Sympathetic denervation thus affects bone resorption within the first few hours after surgery.

Clinical response to the use of the HTR polymer implant in human intrabony lesions.

Fifteen patients, men and women aged 18 to 65 years, were treated for periodontitis. Each patient had at least two bilateral intrabony defects; one was designated the experimental site and the other the control site. Both sites were surgically debrided, but one osseous defect received an HTR polymer implant. Patients were followed for 12 months postoperatively. Maximum pocket reduction occurred within 2 months of initial therapy in both experimental and control sites. Initially, pocket reduction was significantly greater in implanted sites than in control sites, but this difference did not occur subsequently. At the end of the 12 months, pocket reduction responses were similar at control and implanted sites.

Combination therapy in Kala-azar.

128 untreated cases of Kala-azar were divided in 4 equal groups of 32, Group A was treated with Sodium Stibogluconate (SSG) in the dose of 20 mg/kg/body wt. for 30 days. Group B was treated SSG plus allopurinol in the dose of 20 mg/kg/body wt. orally in divided dosage for 30 days. Group C received SSG plus Ketoconazole 600 mg orally in divided dosage for 30 days. Group D in addition to SSG also received levamisole in single oral daily dose of 13 mg/kg/body wt. for 30 days. Response of Group B, C and D was compared to Group A. Results from this study revealed combination of allopurinol with SSG to be statistically not superior to SSG alone.

Direct agglutination test for early diagnosis of Indian visceral leishmaniasis.

In a prospective study, 80 cases of fever with hepatosplenomegaly, anemia and leucopaenia coming from the hyperendemic zones for visceral leishmaniasis of North-Bihar, India were screened and subjected to bone marrow or splenic puncture for demonstration of Leishman-donovan bodies (LDB) and DIRECT AGGLUTINATION TEST (DAT) with antigen prepared by Harith et al. 59 cases were confirmed for Visceral Leishmaniasis (VL) by demonstration of LDB in which DAT was also positive in different titres ranging from 1:1600 onwards. Out of 21 cases in which the bone marrow was negative for parasite, DAT was positive in 10 cases. 8 Out of 10 cases responded to WHO regimen of treatment with sodium stibogluconate (SSG). Remaining two cases who did not respond to this therapy became positive for parasites on subsequent splenic aspirate. They were treated with pentamidine isethionate and were cured. 11 out of 80 cases showing a titre of 1:400 or lower in DAT, 6 proved to be cases of enteric fever and 5 of malaria. Thus DAT using Harith's antigen was found to be 100% sensitive and specific in detection of early cases of Indian VL.

Randomised controlled trial of aminosidine (paromomycin) v sodium stibogluconate for treating visceral leishmaniasis in North Bihar, India.

OBJECTIVES: To assess the efficacy and tolerability of aminosidine compared with sodium stibogluconate for treating visceral leishmaniasis. DESIGN: Randomised, unblinded, controlled trial with 180 day follow up. SETTING: Kala-Azar Research Centre, Brahmpura, Muzaffarpur, Bihar, India. SUBJECTS: People of either sex aged 6-50 years with symptoms and signs suggestive of visceral leishmaniasis (fever, loss of appetite, enlarged spleen) with leishmania amastigotes detected in Giemsa stained aspirates of spleen or bone marrow. INTERVENTIONS: Aminosidine at three daily doses (12, 16, and 20 mg/kg) for 21 days and sodium stibogluconate 20 mg/kg/day for 30 days. MAIN OUTCOME MEASURES: Laboratory measures of efficacy: parasite count, haemoglobin concentration, white cell count, platelet count, serum albumin concentration. Clinical measures of efficacy: spleen size, fever, body weight, and liver size. Measures of safety: liver and renal function tests, reports of adverse events. RESULTS: Of the 120 patients enrolled (30 per treatment arm), 119 completed treatment and follow up. Cure at end of follow up was achieved in 23 (77%), 28 (93%), and 29 (97%) patients treated with 12, 16, and 20 mg aminosidine/kg/day respectively, and in 19 (63%) patients given sodium stibogluconate. At 16 and 20 mg/kg/day, aminosidine was significantly more active than sodium stibogluconate in both clinical and laboratory measures of efficacy. No significant clinical or laboratory toxicity occurred in any treatment group. CONCLUSIONS: A 21 day course of aminosidine 16 or 20 mg/kg/day should be considered as first line treatment for visceral leishmaniasis in Bihar.

Changes in gonadotrophin, lipid and cholesterol levels during annual reproductive cycle in the freshwater teleost, Cirrhinus mrigala (Ham.).

Pituitary gonadotrophin started increasing with gonadosomatic index from preparatory phase but increase in serum gonadotrophin became evident from prespawning phase. However, their peaks were noticed in spawning phase. During other phases of reproductive cycle their values were low except for female pituitary gonadotrophin potency which remained comparatively high during postspawning phase. An increase in ovarian and decrease in testicular lipid started from preparatory phase which continued till spawning phase. Liver lipid in either sex decreased during prespawning and spawning phases. Lipid level was low in ovary but was high in testis and liver during the remaining period of the year. Liver cholesterol was low during spawning and high in prespawning phase whereas serum cholesterol was low during prespawning and spawning phases and remained high during other and peak was recorded during spawning phase. Testicular cholesterol was low during preparatory and postspawning phases and high during the rest of the period of reproductive cycle.

First ovarian maturity and recovery response in common carp, Cyprinus carpio after exposure to carbofuran.

The present study embodies the observations on duration (45, 75 and 105 days) dependent effects of toxicologically low concentration (16 ppm) of a carbamate pesticide, carbofuran, on first ovarian maturity with correlative recovery response by withdrawal of treatment for 30 days in virgin yearlings of common carp, Cyprinus carpio var. Communis. In control fish histological organization of ovary and changes in gonado-somatic index (GSI) revealed the initiation of oocyte differentiation and development at the commencement of experiment in December-January and peak of maturation with fully grown yolky oocytes was attained by the time of termination of experiment in the end of March. In carbofuran exposed groups ovaries exhibited retardation of development and growth evident by significantly decreased GSI and predominace of immature oocytes passing through nuclear-cytoplasmic degeneration of variable magnitude. Severity of degenerative changes was more pronounced with prolonged duration. In recovery group ovaries had experienced regeneration as clued by the presence of oocytes of differential stages including vitellogenic and fully grown oocytes. In liver histopathological changes coupled with significant decrease in hepato-somatic index (HSI) attributed hepatic inactivation in all exposure groups whereas in recovery group an increase in HSI value with marked sign of hepatic reactivation was evidenced. No significant change in values of condition (CF) and somatic condition (SCF) factors substantiated the over all normal health of fish. Lipid and water content of ovary, liver and muscle registered the significant decrease at 105th day of exposure with concomitant augmented value for recovery group. These observations suggest that the commercial formulation of carbofuran, even at this toxicologically low concentration, is equally capable of affecting the first ovarian maturity with associated physiological implications but with reversibility in toxicity by withdrawal of pesticide exposure attributing regeneration in affected organs in due course time.