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Biomedical subjects

Y Singh

Publications and source records attributed to Y Singh.

At least 55 records · Page 3Linked to original sources
PubMed1998

Effect of prenatal diazepam, phenobarbital, haloperidol and fluoxetine exposure on foot shock induced aggression in rats.

Different groups of pregnant rats were treated with diazepam (10 mg/kg), phenobarbital (10 mg/kg), haloperidol (0.1 mg/kg), fluoxetine (10 mg/kg) and vehicle (normal saline) intraperitoneally once a day during gestation days 13 to 21. After birth these pups were culled to 8 pups/dam and foster-nursed by lactating mothers for 3 weeks and were reared in colony cages thereafter. Sex and weight matched pairs of rat offsprings were subjected to foot shock induced aggression test at 8 weeks of age. Two parameters of aggressive behaviour were recorded namely, the latency to fight and total number of fighting bouts. The results indicate that prenatal exposure to diazepam, phenobarbital, haloperidol and fluoxetine caused significantly enhanced aggression in terms of number of fighting bouts.

Aggression↗
PubMed1998

Isolation and characterization of Salmonella enterica Weltevreden cytotoxin.

Cytotoxin of Salmonella enterica subspecies enterica serovar Weltevreden (BM-1643), isolated from buffalo meat, was purified and characterized physicochemically and immunologically. Cell-free culture supernatant (CFCS) of the organism showing marked cytotoxicity to Vero cells and least enterotoxicity to rabbit ligated ileal loop (RLIL) model, was salt precipitated with ammonium sulphate (60% saturation level) and dialysed. Precipitated dialysed preparation (60% PDP) when filtered through Sephadex G-100 column yielded two peaks, of which second peak (SG-100 SP) contained the cytotoxic activity. Upon filtration of SG-100 SP through SG-200 column, three peaks were obtained. Second peak (SG-200 SP), which was cytotoxic, yielded a single protein band of approximately 60-70 kDa in polyacrylamide gel electrophoresis and 3 protein bands of lower, molecular weight (13.5-56 kDa) in SDS-PAGE analysis. Cytotoxic preparation was maximally active at pH 7 to 8. On heating above 60 degrees C, cytotoxicity decreased gradually with insignificant activity left after treatment at 121 degrees C (15 min). Cytotoxin was inactivated by treatment with trypsin and protease but not by papain or lipase enzymes. It was immunogenic in rabbit and antiserum neutralized the cytotoxicity of cytotoxic preparations of homologous as well as heterologous Salmonella serovars.

Animals↗
PubMed1997

Characterisation of virulence factors of Serratia strains isolated from foods.

Out of 21 Serratia strains isolated from fresh juice and fish samples, five S. marcescens and two S. rubidaea caused lethality in mice on parenteral inoculation, but none through oral feeding. Three S. marcescens and one S. rubidaea produced heat-labile enterotoxin, detectible with the rabbit ligated ileal loop test, the mouse foot pad test and the vasopermeability factor test. Cell free culture filtrate (CFCF) of two enterotoxigenic S. mearcescens strains induced cytotoxic effect on a monolayer of Vero-cells, but CFCF of other enterotoxigenic strains could only induce cytotonic changes in Vero-cells. All strains possessed fimbriae type 3 while, only pathogenic strains had type 4 fimbriae and a colonization factor. All pathogenic Serratia strains were agglutinated at comparatively lower salt concentrations than non pathogenic strains (< 1.3 M), and had multiple drug resistance. Their public health significance is also discussed.

Animals↗
PubMed1997

Evaluation of various diagnostic techniques for Trypanosoma evansi infections in naturally infected camels.

One hundred and eight camels (Camelus dromedarius) from Trypanosoma evansi endemic areas of the Thar Desert of Rajasthan State, India, were evaluated by various diagnostic tests including parasitological tests (wet blood film-WBF, stained thick blood film), chemical test (mercuric chloride), biological test (mouse subinoculation-MSI), and immunodiagnostic tests based on antibody detection (double immunodiffusion test-DID, card agglutination test-CATT), antigen detection (double antibody sandwich enzyme linked immunosorbent assay-Ag-ELISA). Of the tested camels 49 were found infected using the WBF of which nine gave false negative results with the mercuric chloride test. The efficacy of MSI was 87.03 percent, while the mercuric chloride test was 60.18 percent efficient. The diagnostic efficacy of CATT (72.22 percent) was found to be much better than DID (28.70 percent). Ag-ELISA was 86.11 percent efficient in detecting trypanosomal antigens. A good correlation was found between the positive results obtained by wet blood film, CATT and Ag-ELISA. It was inferred that CATT can be used to study the seroprevalence of T. evansi with great ease, however, trypanosome antigen detection may give a more accurate idea of the prevalence of T. evansi in an endemic area.

Agglutination Tests↗
PubMed1997

Inhibition of tumor growth and metastasis by angiogenesis inhibitor TNP-470 on breast cancer cell lines in vitro and in vivo.

Antitumor and antimetastatic activity of the angiogenesis inhibitor O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), a semisynthetic analogue of fumagillin, was evaluated in breast cancer cell lines. In an in vitro MTT assay, after 72 hrs continuous exposure to TNP-470, growth inhibition was observed in all seven cell lines of murine (JYG-A, JYG-B, DD-762, and BALB/c-MC) or human (KPL-1, MDA-MB-231, and MKL-F) origin, in which the 50% inhibitory concentrations (IC50) at 72 hrs treatment were 4.6, 4.4, 4.6, 10.1, 35.0, 25.3, and 33.4 micrograms/ml, respectively. In an in vivo assay using JYG-A, JYG-B, KPL-1, and MDA-MB-231 cells by orthotopic (right thoracic mammary fat pad) transplantation in female nude mice, TNP-470 at 30 or 50 mg/kg body weight was injected s.c. every other day from the day of tumor cell inoculation until the end of the experiment. The inhibitory effect on primary tumor growth was obtained in all four cell lines in a dose-dependent manner. In the 50 mg/kg TNP-470-treated group, the reductions in tumor weight of the JYG-A, JYG-B, KPL-1, and MDA-MB-231 cells with respect to the controls were 50%, 30%, 4%, and 49%, respectively. Metastasis was seen in the JYG-A, JYG-B, and KPL-1 cells. The numbers of mice bearing pulmonary metastases of JYG-A and JYG-B cells and regional axillary lymph node metastases of KPL-1 cells were reduced, and TNP-470 at the 50 mg/kg dose to KPL-1 cells significantly reduced lymph node metastases compared with the control. Although the weight gain was retarded in the TNP-470-treated mice, weight loss was not seen. TNP-470 was highly effective in the treatment of breast cancer cells. These results suggest that the clinical use of TNP-470 may be a promising treatment for breast cancer patients.

Animals↗
PubMed1997

Effect of prenatal haloperidol administration on anxiety patterns in rats.

Haloperidol (0.1 mg/kg, i.p.) treatment was given from day 12 to 20 of gestation to pregnant rats, this being the critical period for neural development in this species. The pups born were subjected to open-field exploratory behaviour, tunnel-board exploratory behaviour, elevated zero-maze and elevated plus maze behaviour tests at 7-8 weeks of age. The results indicate that prenatal haloperidol treatment induces a significant increase in open-field ambulations and rearings, decrease in scratching and licking/washing behaviours whereas grooming and faecal droppings remain unchanged. Significantly reduced activity in the centre and increased activity in the periphery of the tunnel board was noted. These suggest presence of anxiety in these animals. Significant anxiogenic behavioural patterns were also observed on elevated zero-maze and plus-maze in the prenatally haloperidol treated offsprings. The results suggest that prenatal exposure of haloperidol leaves a lasting effect on offsprings resulting in hyper-emotional responsiveness and anxiety state.

Animals↗
PubMed1996

Inhibitory Effect of the Angiogenesis Inhibitor O-(Chloroacetyl-carbamoyl)fumagillol(TNP-470) on Tumor Growth and Metastasis of Murine Mammary Tumor Cell Line (JYG-B)Inoculated in Female Nude Mice.

The effect of the potent angiogenesis inhibitor O-(chloroacetyl-carbamoyl) fumagillol(TNP-470), a semisynthetic analogue of fumagillin on tumor growth and metastasis was studied using murine mammary tumor cells (JYG-B) inoculated into female nude mice. Injection of TNP-470 at 10 and 30 mg/kg doses, 3 times a week, until the end of the experiment (41 days)inhibited the s.c.inoculated primary tumor growth in a dose-dependent manner (45%and 65% in volume, respectively). TNP-470 at 30 and 60 mg/kg doses, 3 times a week, until the end of the experiment (35 days), reduced the weight of the i.p. inoculated total tumor mass of abdominal dissemination in a dose-dependent manner (54% and 76%, respectively). Pulmonary metastasis was found in all mice but TNP-470 significantly reduced the lung weight reflecting the total volume of the metastatic foci, and the numbers of metastatic foci in the liver and kidneys was reduced by TNP-470 treatment. These findings show that the angiogenesis inhibitor (TNP-470)has a strong inhibitory effect on tumor grwoth and metastasis in vivo.

Journal Article↗
PubMed1996

Angiosarcoma of the Breast: Immunohistochemical Demonstration of Steroid Receptors and Literature Review.

A 44-year-old female with bilateral angiosarcoma of the breast with multiple skin metastases is reported. She presented with a 6x8 cm and 1x1 cm mass in her left and right breasts, respectively, and multiple skin lesions around the chest. Histologically, the excised tumor was diagnosed as angiosarcoma of the breast with skin metastasis. Tumor cells were immunohistochemically positive with factor VIII-related antigen, CD34, alpha-smooth muscle actin and vimentin, and positive with UEA-I lectin. The majority of tumor cell nuclei expressed progesterone receptor (PgR) protein but estrogen receptro (ER) protein and androgen receptor (AR) protein were nagative. According to the literature, angiosarcoma of the breast mostly affects females in their fourth (33%) and fifth (40%) decades (mean age, 39.5 +/- 10.7 years). The survival rate depends on tumor size and tumor differentiation. The 3-year survival rate is 86% in patients with a tumor size less than 5 cm, and 90% in well-differentiated group I lesions.

Journal Article↗
PubMed1996

Expression and purification of anthrax toxin protective antigen from Escherichia coli.

Anthrax toxin consists of three separate proteins, protective antigen (PA), lethal factor (LF), and edema factor (EF). PA binds to the receptor on mammalian cells and facilitates translocation of EF or LF into the cytosol. PA is the primary component of several anthrax vaccines. In this study we expressed and purified PA from Escherichia coli. The purification of PA from E. coli was possible after transporting the protein into the periplasmic space using the outer membrane protein A signal sequence. The purification involved sequential chromatography through hydroxyapatite, DEAE Sepharose CL-4B, followed by Sephadex G-100. The typical yield of purified PA from this procedure was 500 microg/liter. PA expressed and purified from E. coli was similar to the PA purified from Bacillus anthracis in its ability to lyse a macrophage cell line (J774A.1). The present results suggest that a signal sequence is required for the efficient translocation of PA into E. coli periplasmic space.

Amino Acid Sequence↗
PubMed1996

Effect of ethanol on esophageal cell proliferation and the development of N-methyl-N'-nitro-N-nitrosoguanidine induced-esophageal carcinoma in shrews.

The effect of ethanol (EtOH) on esophageal cell proliferation and the development of esophageal cancers induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in shrews were investigated. Sequential histological examination was done, and cell proliferation was assessed by BrdU labeling. At 5 weeks of age, animals were given tap water, 2% EtOH, 50 ppm MNNG, or 50 ppm MNNG plus 2%, 5% or 10% EtOH in the drinking water. Administration of 10% and 5% EtOH simultaneously with MNNG caused death in 40% (10/25) within 4 days and in 20% (6/30) within 7 days respectively, whereas other treatment were well tolerated with no sudden deaths. Administration of 2% EtOH for 30 weeks caused a 2-fold increase, and that of MNNG caused a 4.5-fold increase in the proliferation index of the basal cells of the esophagus compared with control shrews, and MNNG plus 2% EtOH caused a 5.5-fold increase. In MNNG-treated shrews, with or without 2% EtOH administration, sequential histological examination of esophageal tissue revealed a similar change; dysplasia appeared at 30 weeks of age, squamous cell carcinoma occurred at 35 weeks of age, and the depth of invasion extended to adventitia at 45 weeks of age. These finding indicate that treatment with 2% EtOH promoted the proliferation of esophageal basal cells but did not alter the tumor induction period and did not have tumor-promoting activity. EtOH per se was not carcinogenic; no tumors were seen in shrews not administered MNNG.

Animals↗
PubMed1996

Involvement of prostaglandins in the down-regulation of allergic plasma leakage observed in rats undergoing pleural eosinophilia.

1. Recent evidence has implicated eosinophils in the inhibition of allergen-induced rat pleurisy, but the mechanism of this negative modulation is not completely understood. This study was undertaken in order to define the potential role of prostaglandins in this phenomenon. 2. Wistar rats were actively sensitized by subcutaneous injection of a mixture of ovalbumin and AI(OH)3 and challenged with an intrapleural (i.pl.) injection of ovalbumin (12 micrograms/cavity) 14 days later. 3. Analysis of the pleural fluid effluent revealed a massive mast cell degranulation and plasma protein extravasation 4 h post-challenge. We confirmed that concurrently with selective pleural fluid eosinophilia caused by platelet-activating factor (PAF), the pleural cavity became hyporesponsive to allergen-induced protein exudation and to the parallel reduction in the number of intact mast cells. 4. These hyporesponsive animals presented a significant augmentation in the pleural effluent level of prostaglandin E2 (PGE2), which increased with increasing numbers of eosinophils in the pleural cavity. Furthermore, pretreatment with either indomethacin or aspirin failed to modify allergen-induced exudation but reversed the exudatory hyporesponsiveness associated with eosinophil recruitment. 5. Allergic exudation was clearly down-regulated by the following pretreatments: (i) PGE2 (10 micrograms/cavity, i.pl.) plus rolipram (40 micrograms/cavity, i.pl.), (ii) misoprostol (200 micrograms kg-1, p.o.) or (iii) dibutyryl cyclic AMP (80 micrograms/cavity, i.pl.). 6. We conclude that prostaglandins may be implicated in the eosinophil-mediated inhibition of allergic pleurisy, probably acting via cyclic AMP signalling pathway activation.

Animals↗
PubMed1996

Trout liver slices for metabolism and toxicity studies.

Aquatic species are increasingly used in metabolism and toxicity studies, both from the perspective of potential for chemical exposure and usefulness as nonmammalian model systems. In the present study, trout liver slices were compared with freshly isolated trout hepatocytes with regard to metabolic capabilities and biochemical indices of cell health. Liver slices were also used to discern toxicant-induced changes in liver cell histology. Levels of ATP and glutathione were similar between liver slice and isolated hepatocyte preparations. The cytochrome P450-dependent rate of formation of biphenyl metabolites was 0.48 +/- 0.04 nmol/min/mg protein in slices and 0.43 +/- 0.06 nmol/min/mg protein in isolated cells. 7-Ethoxycoumarin metabolism was also comparable between preparations (1.36 vs. 1.22 nmol/min/mg protein). For conjugative metabolism, glucuronidation of 7-hydroxycoumarin or 1-naphthol did not differ in the two in vitro systems. However, neither slices nor isolated hepatocytes sulfated 7-hydroxycoumarin, whereas 1-naphthylsulfate represented as much as 20% of total 1-naphthol metabolites in both preparations. Histological evaluation of control liver slices after a 24-hr incubation indicated only minor changes. Response to the hepatotoxicants allyl formate and allyl alcohol was evaluated in slices only. Both compounds, after a 4-hr treatment and at concentrations between 0.1 and 1.0 mM, caused extensive depletion of glutathione, but ATP levels were unchanged. Histopathological damage was seen in slices incubated for 24 hr with either toxicant, but was most pronounced with allyl alcohol. These data indicate that liver slices are an excellent in vitro model for metabolism and toxicity studies in aquatic species.

1-Propanol↗
PubMed1996

Behavioural effects of prenatal diazepam administration on anxiety patterns in rats.

Diazepam (10 mg/kg, ip) treatment was given from day 13 to 20 of gestation to pregnant rats, this being the critical period for neural development in this species. The pups born were subjected to open-field exploratory behaviour, tunnel-board exploratory behaviour, elevated zero maze behaviour and social interaction tests at 8-9 weeks of age. The results indicate that prenatal diazepam treatment induces a significant increase in open-field ambulation, grooming, scratching and licking/washing, whereas rearing and faecal dropping remain unchanged. Significant reduction in tunnel-board exploratory activity, activity on zero-maze and social interaction were also observed in the prenatally diazepam treated offsprings. The results suggests that prenatal interference in the form of diazepam leaves a lasting imprint on offsprings resulting in hyper-emotional responsiveness and anxiety state.

Animals↗