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Biomedical subjects

B Singh

Publications and source records attributed to B Singh.

At least 415 records · Page 23Linked to original sources

Novel and potent adenosine 3',5'-cyclic phosphate phosphodiesterase III inhibitors: thiazolo[4,5-b][1,6]naphthyridin-2-ones.

The transformation of 3-bromo-1,6-naphthyridin-2(1H)-ones 8 to thiazolo[4,5-b][1,6]naphthyridin-2(1H)-ones 12 resulted in a 2-9-fold increase in cAMP phosphodiesterase (PDE) III inhibitory potency. Unlike the secondary binding sites on the cAMP PDE III isozyme which interact with the methyl group of milrinone (2) and CI-930 (4), the site which interacts with the 5-substituents of 1,6-naphthyridin-2(1H)-ones and the 8-substituents of thiazolo[4,5-b][1,6]naphthyridin-2(1H)-ones 12 is able to accommodate a diverse group of substituents which have different steric and electronic requirements.

3',5'-Cyclic-AMP Phosphodiesterases↗

Localization of epidermal growth factor (EGF) receptor in the rat corpus luteum, and EGF and transforming growth factor-alpha stimulation of luteal cell steroidogenesis in vitro.

Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) have potent mitogenic effects on granulosa and theca cells. However, their effects on steroidogenesis by these cells is controversial, and there is limited information regarding their effects on luteal cell steroidogenesis. The present study investigated the cellular distribution of the EGF receptor (EGF-R) in the rat corpus luteum (CL) by immunocytochemical staining, and the effects of EGF and TGF-alpha on progesterone and 20 alpha-dihydroprogesterone (20 alpha-OH-P) production in cultures of luteal cells. Using a primary antibody directed against the human EGF-R peptide, specific EGF-R staining was obtained in the CL. Both small and large luteal cells had EGF-R staining. In initial cell culture experiments, treatment of freshly isolated luteal cells with EGF or TGF-alpha (0.5-50 ng/ml) for 24 h had no effect on progesterone and 20 alpha-OH-P accumulation. Addition of LH (250 ng/ml) alone caused a 3.5-fold increase in both progestins, but co-treatment with EGF or TGF-alpha produced no further enhancement of progestin accumulation. However, when cells were seeded overnight and the attached cells were washed prior to growth factor treatment for 3 days with media change every 24 h, both EGF and TGF-alpha caused dose-dependent increases in progesterone accumulation/24 h period (up to 2-fold at 50 ng/ml growth factor) on days 1 and 2 but not day 3 of treatment. 20 alpha-OH-P accumulation was similarly stimulated (up to 2.5-fold) by EGF and TGF-alpha under these conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

20-alpha-Dihydroprogesterone↗

Characterization of MEK1 phosphorylation by the v-Mos protein.

Activation of MAP kinase/Erk Kinase (MEK) via direct phosphorylation by Mos may be crucial for cellular transformation by the activated c-mos or v-mos gene. Recent studies on a number of different protein kinases showed that phosphorylation within a subdomain of the catalytic domain may represent a common mode of activation. In this regard, activation of MEK1 by Raf involves phosphorylation of serine residues 218 and 222. Here we show that recombinant kinase-inactive MEK1 is phosphorylated by v-Mos with equal efficiency at both Ser 218 and Ser 222 in vitro. Tryptic phosphopeptide analysis of glutathione-S-transferase (GST)-MEK1 K97R and its alanine-for-serine mutants indicated that Ser 222 is the preferred phosphorylation site. Wild-type GST-MEK1 was phosphorylated at the same sites but contained a significantly lower amount of doubly phosphorylated species then its K97R kinase-inactive mutant. The ratio of GST-MEK1 species phosphorylated at two serines to those phosphorylated at one serine was similar in auto-phosphorylated and v-Mos-phosphorylated GST-MEK1. Consistent with the in vitro data, phosphopeptide mapping of MEK1 immunoprecipitated from mos transformed cells showed an increased amount of singly phosphorylated phosphopeptide compared to nontransformed cels. MEK1 was found to be more highly activated in NIH3T3 cells transformed by an activated c-mos or v-mos gene than in cells growing normally in medium containing serum. Our data indicate that Mos activated MEK1 in vitro as well as in vivo by phosphorylating Ser 222.

3T3 Cells↗

Implementation of the patient self-determination act in a hospital setting. An initial evaluation.

BACKGROUND: The Patient Self-Determination Act aims to enhance patient awareness of advance directives by requiring health-care institutions to ask patients whether they have advance directives and to inform patients of their rights to prepare these documents. We investigated the following: (1) compliance of the hospital staff with implementing this act, (2) the effects of this act on the extent to which patients discuss and prepare advance directives, and (3) variables that might influence patient discussions on advance planning and preparation of advance directives. METHODS: We surveyed 219 patients from a university hospital that implemented a nurse-dependent advance directive program. We also conducted a telephone interview with 57% of these patients at least 6 months after hospital discharge. RESULTS: Nurses asked 70% of the patients about the existence of an advance directive and of these patients, only 57% remembered the inquiry. Only 57% of the patients received the brochure on advance directives and of these patients, only 55% read the brochure. Only 2% of the patients requested to receive additional information on advance directives. Less than one quarter of the patients had discussions on advance planning while in the hospital and of those patients who were contacted within 6 months after hospital discharge, 39% had discussions on advance planning and 15% prepared an advance directive. Race was an independent predictor for hospital discussions, and educational level was an independent predictor for discussions and preparation of advance directives after hospital discharge. CONCLUSIONS: To enhance the effectiveness of a nurse-dependent advance directive program, hospitals may need (1) to strengthen the quality of the patient-nurse encounter in which the issue of advance directives is raised to more effectively promote patient interest, discussions, and preparation of advance directives and (2) to account for the social diversity of their patient population.

Adult↗

Surface coat of sheep pulmonary intravascular macrophages: reconstitution, and implication of a glycosyl-phosphatidylinositol anchor.

BACKGROUND: Pulmonary intravascular macrophages (PIMs) of sheep have a globular surface coat that facilitates endocytosis of tracer particles and Escherichia coli lipopolysaccharide, and is disrupted by the heparin and Brefeldin A treatments. The present study investigated the in vivo dynamics of the coat globules following heparin-mediated removal, and the mechanism of globule organization on the plasma membrane of PIMs in vitro. METHODS: Sheep were administered heparin at a dose of 50 IU/kg body weight IV, and euthanised at 30 min, 3, 6, 12, 48, and 120 hr (n = 2 for each treatment) after the treatment. Control sheep (n = 2) were injected with normal saline solution. The tissues were processed for an ultrastructural examination and acid phosphatase (ACPase) cytochemistry. Heparin-treated lungs were subjected to morphometric analysis of the coat globules. Lung tissues from normal sheep (n = 2) were incubated with phosphatidylinositol-specific-phospholipase C (PIPLC; 2 IU/ml PBS) in vitro for 30 and 75 min. RESULTS: Heparin study: The ultrastructural and morphometric data showed that the coat globules were removed at 30 min and reconstituted within 48 hr of the treatment. The PIMs showed prominent Golgi complexes associated with secretory vesicles, microtubules, and centriole between 3-12 hr of heparin treatment. Acid phosphatase cytochemistry also demonstrated secretory activity in the Golgi complexes of PIMs during the coat reconstitution. PIPLC study: The coat globules of PIMs were removed in a time-dependent mode by the PIPLC treatment without damage to other cell organelles. CONCLUSIONS: This study demonstrates a time-dependent reconstitution of the coat of PIMs in conjunction with secretory activity following heparin-mediated removal, probably through sequestration of the globules from blood. This ability is of functional significance as the coat mediates particle endocytosis by the PIMs. The results also suggest the presence of a glycosyl-phosphatidylinositol (GPI) anchor in tethering of globules on the plasma membrane of PIMs to offer a structural basis for their integrity in pulmonary vascular flow.

Animals↗

Carbon-laden macrophages in pleural fluid of crack smokers.

Carbon-laden macrophages in bronchoalveolar lavage have been noted to be associated with a history of crack smoking. We report herein the finding of carbon-laden macrophages in four cytological preparations of pleural fluid from two crack smokers. The etiology of the two patients' pleural effusions differed; neither had a bronchopleural fistula. Patient 1 had AIDS, Pneumocystis carinii pneumonia, and Kaposi's sarcoma of the right lung with an associated bilateral pleural effusion. Patient 2 was HIV seropositive, had pulmonary tuberculosis, hepatitis A, B, and C, cardiomyopathy, pulmonary embolism, and bilateral pleural effusions, the latter of which were probably due to cardio-pulmonary dysfunction. An additional two crack smokers with pleural effusions due to malignancy, one primary pulmonary adenocarcinoma and the other metastatic melanoma, did not have carbon-like material in their pleural fluid cytology. We hypothesize that intracellular accumulation of carbonaceous material in the lung parenchyma and pleural space occurs when normal clearance mechanisms are overwhelmed.

Acquired Immunodeficiency Syndrome↗

Epidermal growth factor and its receptor gene expression and peptide localization in porcine ovarian follicles.

The present study was undertaken to determine the expression of genes for epidermal growth factor (EGF) and its receptor (EGF-R) in various components of medium-sized porcine ovarian follicles by reverse transcription-polymerase chain reaction (RT-PCR), and to localize their peptides during folliculogenesis by immunocytochemistry. A strong band for EGF mRNA transcript was detected in the oocyte, whereas the signal in cumulus, granulosa, and theca cells was very weak but detectable. In contrast, a very strong EGF-R mRNA signal was observed in cumulus, granulosa, and theca cells, whereas the signal in the oocyte was very weak. EGF peptide was localized in the oocyte, cumulus, and granulosa cells of all stages of follicle. In the oocyte, the intensity of immunostaining was more pronounced in primordial and primary follicles, compared to atrial follicles. In large antral follicles, immunostaining was pronounced in granulosa cells, whereas theca cells showed little or no detectable staining for EGF. EGF staining was also observed in the cumulus and granulosa cells of follicles undergoing atresia. EGF-R immunostaining was observed in the oocytes of primordial and primary follicles, and in cumulus, granulosa, and theca cells of all stages of follicle, including atretic follicles. In large antral follicles, the intensity of immunostaining was more pronounced in theca cells than in granulosa cells, and the oocyte showed little or no detectable staining. No immunostaining was observed when the primary antibody was replaced with preimmune serum (EGF), or preabsorbed with the control peptide (EGF-R), confirming the specificity of the staining procedures.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

X-ray fluorescence in the assessment of inter-elemental interactions in rat liver following lead treatment.

Energy dispersive X-ray fluorescence technique was employed to study the interactions of lead (50 and 100 mg/kg body wt) with K, Fe, Cu, Zn, Br and Rb in rat liver. Lead was administered orally to rats daily for dosage periods of 1 and 4 months (short and long terms). Hepatic Fe levels were found to increase significantly with the supplementation of low and high doses of lead for both the treatment periods, although the increase was more pronounced following long-term treatment. The levels of hepatic K, Cu and Br were seen to decrease significantly over both time intervals. Moreover, hepatic Rb contents were lowered with the short-term supplementation of low doses of lead. In contrast, Rb and Zn levels were increased when lead was administered for the longer period at both dose levels.

Animals↗

Suitability of full fat and defatted rice brain obtained from Indian rice for use in food products.

The effect of blending different types of rice bran in wheat flour on the rheological, pasting and baking properties is reported. Dough development time and strength as measured through farinograph decreased with the blending of unstabilized full fat and defatted bran; whereas, blending of stabilized full fat and defatted bran improved the dough strength. Pasting properties revealed an increase in gelatinization temperature and decrease in paste viscosities and set-back values with the blending of various types of rice bran. Bread volume and cookie spread decreased with blending of different types of rice bran; however, the decrease was more pronounced with the defatted bran. Muffin volume improved with the blending of rice bran. Stabilized full fat rice bran up to 20% level and unstabilized full fat or stabilized defatted rice bran up to 10% was found suitable in various food products.

Dietary Fats↗

Pathological studies in broiler chicks fed aflatoxin or ochratoxin and inoculated with inclusion body hepatitis virus singly and in concurrence.

Day-old broiler chicks found negative for maternal antibodies against inclusion body hepatitis (IBH) virus by agar gel precipitation test and viral antigen in cloacal swabs by dot enzyme immunoassay were divided into 6 groups of 20 chicks each. Group A was fed aflatoxin B1 at 1.25 ppm from 3 to 38 days of age; group O was fed ochratoxin A at 0.5 ppm from 3 to 38 days of age; group V was inoculated with 1 ml of IBH virus of titre log10 6.5 EID50 per 0.2 ml. Groups AV and OV were given aflatoxin B1 and ochratoxin A, respectively, and also infected with the virus. Group C served as control. There was mild enlargement and paleness of the liver up to 18 days post inoculation in group V; there were no lesions in group A; and there was gradual enlargement of the kidneys from 10 days post feeding of mycotoxin onwards in group O. In the combined groups AV and OV the gross lesions were slightly more severe. In group V, varying degrees of degenerative histopathological changes, congestion and haemorrhages were seen particularly in the liver, followed by the kidneys, bursa, spleen, myocardium and lungs, along with intranuclear inclusion bodies in the hepatocytes, mostly in the early stages of infection. Similar microscopic changes, but without inclusion bodies, were seen in groups A and O and the changes were pronounced in the later stages. In group O, the kidney lesions were more pronounced than the liver lesions. In the concurrently infected groups, AV and OV, the changes were similar but slightly more marked than in the corresponding individual groups. Inclusion bodies in hepatocytes were more frequent, more prominent and appeared earlier in the concurrent groups.

Adenoviridae Infections↗

Pathogenicity of quail's inclusion body hepatitis virus (avian adenovirus-1) for Japanese quails and broiler chicks.

Quail (Coturnix coturnix japonica) and broiler (Gallus domesticus) chicks were inoculated experimentally with IBH virus (avian adenovirus-1) derived from quails to determine its pathogenicity. Quail chicks were inoculated by the intraperitoneal route at 3, 4, 5, 6 or 7 weeks of age. Lesions were encountered most frequently in the liver, kidneys and lungs. These included pale, swollen and mottled liver, swollen nephrotic kidneys, and congested and pneumonic lungs. The lesions were severe in birds inoculated at 5 weeks of age. Large basophilic intranuclear inclusion bodies were seen in hepatocytes and occasionally in the renal epithelium. The results showed that this isolate is pathogenic for quails above 3 weeks of age. Broiler chicks were inoculated at 4 weeks of age by the intraperitoneal route. The lesions produced in these chicks were similar to those of adenovirus-induced inclusion body hepatitis. Viral antigen was also demonstrated by dot-ELISA in suspension of liver tissue from both quail and broiler chicks.

Adenoviridae Infections↗

A comparison of intravenous ranitidine and omeprazole on gastric volume and pH in women undergoing emergency caesarean section.

We have compared the effect of intravenously administered omeprazole and ranitidine on gastric contents in a double-blind study in 80 consecutive women undergoing emergency Caesarean section. When the decision to perform emergency Caesarean section was made, patients were randomly assigned to receive either ranitidine 50 mg or omeprazole 40 mg intravenously. The volume and pH of the gastric contents were measured immediately after tracheal intubation and again before extubation. The gastric pH was found to be higher after omeprazole than after ranitidine immediately after intubation (5.89 +/- 1.46 and 5.21 +/- 1.36 respectively) (P < 0.05) and before extubation (5.97 +/- 1.38 and 5.32 +/- 1.24 respectively) (P < 0.05). However, the gastric volumes were comparable in both the groups. The number of patients with gastric volume > 25 ml and pH < 2.5 were 3 (7.5%) in the ranitidine group and 1 (2.5%) in the omeprazole group after intubation and none in either of the groups before extubation. We conclude that omeprazole 40 mg iv administered at the time of the decision to operate, results in higher gastric pH than ranitidine in obstetric patients undergoing emergency Caesarean section.

Cesarean Section↗

An N-formyl cyclopeptide alkaloid from Zizyphus nummularia bark.

A 13-membered N-formylcyclopeptide alkaloid, nummularine-T, has been isolated from the bark of Zizyphus nummularia and its structure established by spectroscopic and chemical methods. It provides the fourth example of a naturally occurring N-formyl cyclopeptide alkaloid.

Alkaloids↗