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Biomedical subjects

J Singh

Publications and source records attributed to J Singh.

At least 91 records · Page 5Linked to original sources
PubMed2001

Morphine-induced degradation of the host defense barrier: role of macrophage injury.

The effect of morphine on the degradation of the host defense barrier in rats and mice was studied. Mice received either 3 or 11 doses of morphine. Mice receiving 11 doses of morphine showed gram-negative bacteremia and bacterial growth in samples of peritoneal fluid (PF), liver, spleen, kidneys, heart, and lungs; PF and tissue samples from only 1 control mouse showed bacterial growth, and no control mice had bacteremia. Mice receiving 11 doses also had suppressed bone marrow macrophage colony formation. Monocytes and peritoneal macrophages harvested from morphine-treated mice showed greater injury than did those from control mice. Pretreatment of mice with naloxone inhibited morphine-induced macrophage injury and degradation of the host defense barrier. In in vitro studies, morphine attenuated the killing of bacteria phagocytosed by macrophages and also facilitated their escape. This study indicates that morphine-induced monocyte and macrophage injury may be linked to degradation of the host defense barrier.

Analgesics, Opioid↗
PubMed2001

Chromodomain protein Swi6-mediated role of DNA polymerase alpha in establishment of silencing in fission Yeast.

Although DNA replication has been thought to play an important role in the silencing of mating type loci in Saccharomyces cerevisiae, recent studies indicate that silencing can be decoupled from replication. In Schizosaccharomyces pombe, mating type silencing is brought about by the trans-acting proteins, namely Swi6, Clr1-Clr4, and Rhp6, in cooperation with the cis-acting silencers. The latter contain an autonomous replication sequence, suggesting that DNA replication may be critical for silencing in S. pombe. To investigate the connection between DNA replication and silencing in S. pombe, we analyzed several temperature-sensitive mutants of DNA polymerase alpha. We find that one such mutant, swi7H4, exhibits silencing defects at mat, centromere, and telomere loci. This effect is independent of the checkpoint and replication defects of the mutant. Interestingly, the extent of the silencing defect in the swi7H4 mutant at the silent mat2 locus is further enhanced in absence of the cis-acting, centromere-proximal silencer. The chromodomain protein Swi6, which is required for silencing and is localized to mat and other heterochromatin loci, interacts with DNA polymerase alpha in vivo and in vitro in wild type cells. However, it does not interact with the mutant pol alpha and is delocalized away from the silent mat loci in the mutant. Our results demonstrate a role of DNA polymerase alpha in the establishment of silencing. We propose a recruitment model for the coupling of DNA replication with the establishment of silencing by the chromodomain protein Swi6, which may be applicable to higher eukaryotes.

Base Sequence↗
PubMed2001

Lipid extraction and transport of hydrophilic solutes through porcine epidermis.

The purpose of this study was to investigate the effect of delipidization of the stratum corneum (SC) on the in vitro percutaneous absorption of hydrophilic solutes (i.e. water, urea, and inulin). Fourier transform infrared (FT-IR) spectroscopy was employed to study the extent of delipidization of porcine SC due to chloroform:methanol (2:1) (C:M (2:1)) treatments for various time periods. In vitro percutaneous absorption of [3H] water, [14C] urea, and [3H] inulin were studied through C:M (2:1) treated epidermis in Franz diffusion cells. There was a greater decrease in peak areas of the asymmetric and symmetric C-H stretching absorbances (i.e. increase in lipid extraction) with increasing exposure times of the SC with C:M (2:1). After 40-min treatment, asymmetric and symmetric C-H stretching peak area showed a decrease of 75.9 and 89.9%, respectively. The permeability coefficient of water, urea, and inulin increased with increasing lipid extraction. Enhancement in the permeability coefficient, through 40 min C:M (2:1) treated epidermis in comparison to the control, for water, urea, and inulin was 48.72, 215.65, and 3.90, respectively. Log (permeability coefficient) and log (mol. wt.) for test solutes and leuprolide acetate were found to be inversely related (R(2)=0.9974). In conclusion, this study implies that penetration enhancers that are safe and extract the SC lipids can be selected in order to enhance the percutaneous absorption of polar solutes through the skin.

Animals↗
PubMed2001

Id-1, ITF-2, and Id-2 comprise a network of helix-loop-helix proteins that regulate mammary epithelial cell proliferation, differentiation, and apoptosis.

Mammary epithelial cells proliferate, invade the stroma, differentiate, and die in adult mammals by mechanisms that are poorly understood. We found that Id-1, an inhibitor of basic helix-loop-helix transcription factors, regulates mammary epithelial cell growth, differentiation, and invasion in culture. Here, we show that Id-1 is expressed highly during mammary development in virgin mice and during early pregnancy, when proliferation and invasion are high. During mid-pregnancy, Id-1 expression declined to undetectable levels as the epithelium differentiated fully. Surprisingly, Id-1 increased during involution, when the epithelium undergoes extensive apoptosis. To determine whether Id-1 regulates both proliferation and apoptosis, we constitutively expressed Id-1 in mammary epithelial cell cultures. Id-1 stimulated proliferation in sparse cultures but induced apoptosis in dense cultures, which reflect epithelial cell density during early pregnancy and involution, respectively. To understand how Id-1 acts, we screened a yeast two-hybrid library from differentiating mammary epithelial cells and identified ITF-2, a basic helix-loop-helix transcription factor, as an Id-1-interacting protein. Overexpression of ITF-2 significantly reduced Id-1-stimulated proliferation and apoptosis. We show further that, in contrast to Id-1, Id-2 was expressed highly in differentiated mammary epithelial cells in vivo and in culture. In culture, Id-2 antisense transcripts blocked differentiation. Our results suggest that Id-1, ITF-2, and Id-2 comprise a network of interacting molecular switches that govern mammary epithelial cell phenotypes.

Animals↗
PubMed2001

Global modulation of cellular transcription by human cytomegalovirus is initiated by viral glycoprotein B.

Human cytomegalovirus (HCMV) infection alters the expression of many cellular genes, including IFN-stimulated genes (ISGs) [Zhu, H., Cong, J.-P., Mamtora, G., Gingeras, T. & Shenk, T. (1998) Proc. Natl. Acad. Sci. USA 95, 14470-14475]. By using high-density cDNA microarrays, we show that the HCMV-regulated gene expression profile in fibroblasts does not differ substantially from the response generated by IFN. Furthermore, we identified the specific viral component triggering this response as the envelope glycoprotein B (gB). Cells treated with gB, but not other herpesviral glycoproteins, exhibited the same transcriptional profile as HCMV-infected cells. Thus, the interaction of gB with its as yet unidentified cellular receptor is the principal mechanism by which HCMV alters cellular gene expression early during infection. These findings highlight a pioneering paradigm for the consequences of virus-receptor interactions.

Cytomegalovirus↗
PubMed2001

Effect of menthone on the in vitro percutaneous absorption of tamoxifen and skin reversibility.

The effect of penetration enhancer (i.e., 1, 2, 3 and 5% menthone in combination with 50% ethanol (EtOH)) was investigated on the in vitro percutaneous absorption of tamoxifen, and post-recovery epidermal permeability after removal of the above enhancer. The flux of tamoxifen with menthone in combination with 50% EtOH was significantly greater (P<0.05) than the control (50% EtOH). The flux of tamoxifen increased with increasing concentrations of menthone. The post-recovery flux through enhancer exposed epidermis was significantly decreased (P<0.05) as compared to pre-recovery. However, post-recovery flux of tamoxifen through the enhancer-exposed epidermis did not completely recover to the baseline (i.e., post-recovery flux through phosphate buffered saline, pH 7.4 treated epidermis).

Administration, Topical↗
PubMed2001

Observation of phonon bottleneck in quantum dot electronic relaxation.

Time-resolved differential transmission measurements of self-assembled In0.4Ga0.6As quantum dots clearly indicate a phonon bottleneck between the n = 2 and n = 1 electronic levels. The key to this observation is the generation of electrons in dots where there are no holes so that electron-hole scattering does not mask the bottleneck. We use a simple carrier capture model consisting of two capture configurations to explain the bottleneck signal and offer arguments to rule out other possible sources of the signal.

Journal Article↗
PubMed2001

Lipid extraction and iontophoretic transport of leuprolide acetate through porcine epidermis.

The purpose of this study was to explore the effect of lipid extraction by the simple alkyl acetates of increasing carbon chain lengths (e.g. methyl, ethyl, propyl, butyl, pentyl, hexyl, and octyl acetates) and iontophoresis on the in-vitro transport of leuprolide acetate through porcine epidermis. The extent of lipid extraction from the stratum corneum (SC) by alkyl acetates was studied by Fourier transform infrared (FT-IR) spectroscopy. Ethyl, propyl, pentyl, hexyl, and octyl acetates significantly increased (P < 0.05) the permeability of leuprolide acetate through the epidermis in comparison to the control (epidermis without alkyl acetate treatment). Iontophoresis further increased (P < 0.05) the permeability of leuprolide acetate for all the alkyl acetates studied, when compared to their corresponding passive permeability. Ethyl acetate produced the maximum passive (13.47 microg/cm(2)/h) and iontophoretic (89.79 microg/cm(2)/h) flux among all the alkyl acetates studied. The SC treated with alkyl acetates showed a decrease in peak heights and areas of asymmetric and symmetric C--H stretching absorbances in comparison to untreated SC. A greater percentage decrease in peak heights and areas was obtained by ethyl acetate. Chloroform:methanol(2:1) [C:M(2:1)] was used as a positive control for lipid extraction. Our findings provide evidence that alkyl acetates cause lipid extraction, which leads to an enhancement in the passive and iontophoretic permeability of leuprolide acetate.

Animals↗
PubMed2001

In vitro transdermal iontophoretic transport of timolol maleate: effect of age and species.

Transdermal iontophoresis would be a promising method for the systemic delivery of water soluble and ionic drugs of relatively high molecular size, including peptides. In the present study, the effect of biological parameters such as age of the animal and species variation (rat, rabbit, mouse, guinea pig and human) on the transdermal iontophoretic transport was studied using timolol maleate (TM) as a model drug. The iontophoretic transport of TM across the skins obtained from the rats of different age groups was found to be similar. The results of the present study suggest that the age of the animal (Wistar rats: 1-8 months) did not appear to influence the transdermal iontophoretic transport of TM significantly. The amount of TM transported during iontophoresis (2 h) was significantly different among the different skin species. But the total amount of TM transported up to 24 h (2 h iontophoresis+22 h post-iontophoretic passive diffusion) was not significantly different among the different species studied. The present study provides further evidence that iontophoresis technique reduces the interspecies differences in the transdermal permeation of drugs, which is normally observed in passive diffusion of drugs. However, it must be noted that excised skins have been used in the present study to investigate the role of age and species variation on the iontophoretic transport of TM. The influence of these parameters under in vivo conditions might be different considering the physiological differences in different species and in the animals of different age groups.

Administration, Cutaneous↗
PubMed2001

Molecular cloning and characterization of a zinc finger protein involved in Id-1-stimulated mammary epithelial cell growth.

Id proteins are dominant negative regulators of basic helix-loop-helix transcription factors. Previous work in our laboratory has shown that constitutive expression of Id-1 in SCp2 mouse mammary epithelial cells inhibits their differentiation and induces proliferation, invasion, and migration. Id-1 expression also correlates with the invasive and aggressive potential of human breast cancer cells. However, little is known about Id-1 target genes that are important for regulating normal and transformed breast epithelial cell phenotypes. Now we report the cloning of a novel zinc finger protein, Zfp289, using degenerate primers to specifically amplify cDNAs from Id-1-transfected SCp2 cells. Zfp289 has homology with a yeast zinc finger protein, the GTPase-activating protein Gcs-1, which was initially identified as a gene required for the re-entry of cells into the cell cycle after stationary phase growth. Zfp289 mRNA expression pattern correlates with Id-1 expression in SCp2 mammary epithelial cells under various experimental conditions as well as in the mouse mammary gland at different stages of development. It is predominantly present in the cytoplasm of the cells as evident from green fluorescent protein fusion protein localization. SCp2 mammary epithelial cells with constitutive expression of Zfp289 have a higher S-phase index, compared with control cells, when cultured in a serum-free medium. We conclude that the novel zinc finger protein Zfp289, which may represent the mammalian homologue of Gcs-1, is potentially an important mediator of the Id-1-induced proliferation pathway in mammary epithelial cells.

Amino Acid Sequence↗
PubMed2001

Expression of Fos immunoreactivity in some catecholaminergic brainstem neurons in rats following high-altitude exposure.

This study examined the response of neurons of the cardiorespiratory centers, i.e., the nucleus tractus solitarius and the ventrolateral medulla as well as the area postrema in adult and postnatal rats subjected to high-altitude exposure at 4,000 m and 8,000 m. In adult control rats, sporadic Fos-positive neurons were detected in the above-mentioned areas. On exposure to 4,000 m altitude, the number of Fos-positive neurons was noticeably increased. At 8,000 m, the incidence of labeled cells was markedly increased, with many of them doubly labeled for tyrosine hydroxylase. In postnatal rats, Fos expression was not detected in these areas in either control rats or rats exposed to 4,000 m altitude. Fos-positive cells, however, were observed in the these areas in postnatal rats exposed to 8,000 m. In the latter, tyrosine hydroxylase labeling was observed in some Fos-positive cells in the nucleus tractus solitarius and ventrolateral medulla. In rats killed at 24 hr after exposure to high altitude, Fos expression in both the adult and the postnatal rats was comparable to that in their corresponding control rats. Present results suggest that Fos expression in various brainstem areas was induced by reduced oxygen tension in the ambient air at high altitude. Double labeling of some Fos-positive neurons with tyrosine hydroxylase indicates an increased sympathetic activation, which may be involved in the mediation of cardiorespiratory responses to hypoxia. This, however, was less evident in the postnatal animals. It is possible that the peripheral chemoreceptors or the regulation of autonomic functions is not fully developed in this age group.

Altitude Sickness↗
PubMed2001

Effect of lindane on the growth and metabolic activities of cyanobacteria.

The effect of lindane was studied in Cyanobacteria (Anabaena) by estimating its growth pattern, biomass yield, chlorophyll content, and total starch and protein content. The results reveal that exposure of Anabaena with 0.5, 1.0, 1.5, and 2.0 ppm levels of lindane will have drastic effects on biomass production and photosynthetic rate. Since Cyanobacteria are a primary source of aquatic food web and important biofertilizer for rice cultivation, their protection from residual effects of lindane is essential for enriched soil fertility.

Algal Proteins↗
PubMed2001

Nasal mucociliary clearance in healthy children in a tropical country.

INTRODUCTION: Nasal mucociliary clearance is an important physiological function of nasal cavity that helps in protecting the lower respiratory tract from undesirable organic and inorganic matter including the micro organisms. The study was designed to establish normal mucociliary clearance time in healthy children in a tropical environment. MATERIAL AND METHODS: The study was carried out in 100 randomly selected normal school children aged 4--15 years using saccharin method. The diseases that are known to affect the nasal mucociliary clearance were excluded. The study variables were age and sex. RESULTS: Mean nasal mucociliary clearance time was 5.7+/-2.59 min with no significant difference between males and females. The clearance time was found to be impaired in groups A and B including children of 4--7 and 8--11 years of age respectively. CONCLUSION: Nasal mucociliary clearance is impaired in children of either sex between 4 and 11 years probably due to subclinical adenoiditis. However clearance returns to normal level at the time of puberty, which coincides with adenoids involution.

Adolescent↗
PubMed2001

Distribution of vasoactive intestinal polypeptide, neuropeptide-Y and substance P and their effects on insulin secretion from the in vitro pancreas of normal and diabetic rats.

This study examined the pattern of distribution of vasoactive intestinal polypeptide (VIP), neuropeptide-Y (NPY) and substance P (SP) in the pancreas of diabetic rat to determine whether there are changes in the number and pattern of distribution of these neuropeptides after the onset of diabetes. Moreover, the effect of VIP, NPY and SP on insulin secretion from the pancreas of normal and diabetic rats was also examined. Diabetes mellitus (DM) was induced by a single dose of streptozotocin (STZ) given intraperitoneally (i.p.) (60 mg kg body weight(-1)). Four weeks after the induction of DM, diabetic (n = 6) and normal (n = 6) rats were anesthetized with chloral hydrate and their pancreases removed and processed for immunohistochemistry and insulin secretion. The number of insulin-positive cells in the islets of Langerhans was reduced while that of VIP and NPY increased significantly after the onset of diabetes. The pattern of distribution of VIP, NPY and SP in the nerves innervating the pancreas was similar in both normal and diabetic rats. VIP-evoked large and significant (P < 0.02) increases in insulin secretion from the pancreas of normal and diabetic rats. NPY also induced a marked (P < 0.005) increase in insulin release from pancreatic tissue fragments of normal rat. Stimulation of pancreatic tissue fragments of diabetic rat with NPY resulted in a slight but not significant increase in insulin release. SP induced a large and significant (P < 0.005) increase in insulin secretion from the pancreas of normal rat but inhibited insulin secretion significantly (P < 0.03) from isolated pancreas of diabetic rat. In summary, VIP and NPY can stimulate insulin secretion from the pancreas after the onset of diabetes. The stimulatory effect of SP on insulin secretion is reversed to inhibitory in diabetic rats.

Animals↗
PubMed2001

Regional variations in skin barrier function and cutaneous irritation due to iontophoresis in human subjects.

The effect of saline iontophoresis on skin barrier function and irritation was investigated on three body sites (abdomen, chest and upper arm) in order to select an appropriate site for iontophoretic delivery of drugs. Thirty healthy human volunteers were recruited according to specific entry criteria. Ten subjects, five males and five females, were assigned to each body site group. Skin barrier function and irritation was examined after 4 h of saline iontophoresis at a current density of 0.2 mA/cm(2) on a 6.5 cm(2) area in terms of the measured responses: transepidermal water loss (TEWL), skin capacitance, skin temperature and visual scores. Alterations in TEWL due to iontophoresis were not observed in the upper arm and chest; however, changes in TEWL at the abdomen were observed and returned to baseline 2 h after patch removal. Similarly, changes in capacitance due to iontophoresis returned to baseline (P>0.05) at the three body sites 2 h after patch removal except under the anode at the abdomen (P<0.05). There was a significant increase in skin temperature due to iontophoresis at the anode and the cathode (P<0.05) at the upper arm. Edema was not observed. At patch removal, the erythema score was significantly (P<0.001) elevated in comparison to baseline at the three body sites. Erythema resolved within 24 h except at the chest under the anode, where the erythema score was still higher (P<0.01) than the baseline. Papules appeared in five subjects at the active anode site on the chest. In three of the subjects, these papules did not resolve until 24 h post patch removal. Thus, there was regional variation in the function of the skin and irritation due to iontophoresis. Irritation was greater at the chest than at the abdomen or upper arm.

Abdomen↗
PubMed2001

The relation of the peak Doppler E-wave to peak mitral annulus velocity ratio to diastolic function.

Previous observations (Garcia et al. 1998; Sohn et al. 1997; Nagueh et al. 1997) indicate that mitral annulus velocity may be viewed as a "load-independent" index of filling and that wedge pressure is linearly related to the transmitral flow velocity (V(E)) to mitral annulus velocity (V(MA)) ratio (V(E)/V(MA)) measured at maximum velocity. In healthy subjects, the mean value observed for [V(E)](max)/[V(MA)](max) was 1:0.23 approximately 4. No prior physiologic or physical explanation for the basis of, or for the magnitude of, the ratio has been proposed. We propose a physiologic, model-based, quantitative explanation for these observations and test our simplified model's prediction in an invasive (n = 30) and noninvasive (n = 34) test groups of subjects. The simplified geometric model is based on the known constant volume (within a few percentage points) attribute of the four-chambered heart. Accordingly, left-atrial and left-ventricular volumes reciprocate so that their sum is constant throughout the cardiac cycle. The model predicts that: 1. the ratio (V(E)/V(MA)) is a constant approximately 3 in healthy hearts; and 2. V(E)/V(MA) should be linearly proportional to left ventricular end-diastolic pressure (LVEDP). Model prediction was tested using V(E) and V(MA) echocardiographic data from 34 subjects (noninvasive group), and simultaneous echocardiographic and high-fidelity hemodynamic (LVEDP) data in 30 subjects (invasive group). Excellent agreement was observed between model prediction and observed data. For the noninvasive (healthy) group, [V(E)](max)/[V(MA)](max) = 4.20 +/- 1.11. For the invasive group, [V(E)](max)/[V(MA)](max) was observed to be linearly related to LVEDP, [V(E)](max)/[V(MA)](max) = 0.19 (LVEDP) + 1.54, r = 0.92. Hence, [V(E)](max)/[V(MA)](max) is a legitimate flow-derived index of diastolic function because it is a derivable consequence of the heart's constant-volume pump attribute.

Blood Flow Velocity↗
PubMed2001

Screening of various Swertia species extracts in primary monolayer cultures of rat hepatocytes against carbon tetrachloride- and paracetamol-induced toxicity.

Swertia chirata Buch-Ham. (Gentianaceae), one of the oldest medicinal herbs of India, is a source of the Indian ayurvedic drug 'chirata' used for the treatment of liver disorders and malarial fevers. In this study, eight species of Swertia were collected. Each of the dry whole plant was extracted into methanol, the aqueous extract of which was sequentially extracted into hexane, chloroform and butanol extracts. The extracts were screened for their anti-hepatotoxic activity against carbon tetrachloride (CCl4) and paracetamol (acetaminophen (AAP)) toxicity in primary monolayer cultures of rat hepatocytes. The primary cultures, 2.5 x 10(6) cells /3 ml medium/60 mm collagen-coated plates, were exposed to 2.5 mM CCl4 or 12 mM AAP in the presence or absence of plant extracts (100 microg/ml culture medium). Cells and medium were harvested after 22 h of treatment for the assay of cellular reduced gluthathione (GSH) content and leakage of lactate dehydrogenase as biological end-points of toxicity. Both CCl4 and AAP at the indicated concentrations reduced GSH by almost 50 and 80%, respectively, while the enzyme leakage was almost 15% above the untreated control. Hexane and methanol extracts of most of the species in general offered relatively good protection. The anti-hepatotoxic activity, nevertheless, was evident in all Swertia species against both the toxicants. However, Swertia purpurascens, Swertia chirata, Swertia paniculata and Swertia cordata exhibited better activity compared with other species investigated. In addition, influence of various extracts (10-100 microg/ml medium) was examined on cellular growth of rat Reuber hepatoma cell line H4IIEC3/G-. Except for the butanol extract of S. chirata, no other extracts exerted toxicity in terms of neutral red uptake by the cells.

Acetaminophen↗