Biomedical subjects

S Sinha

Publications and source records attributed to S Sinha.

At least 253 records · Page 14Linked to original sources

PubMed1998

Effect of nutritional status on vecuronium induced neuromuscular blockade.

Based on simple clinical and biochemical parameters of nutritional status, seventy adult patients scheduled for elective surgery under general anaesthesia were categorized as having normal nutrition, mild, moderate or severe malnutrition or obesity. Under anaesthesia, evoked responses on train-of-four nerve stimulation were recorded every 15 seconds on a mechanomyograph. Vecuronium 0.1 mg.kg-1 was used to achieve neuromuscular blockade. Compared with patients having normal nutrition, the time to onset of action was significantly prolonged in the moderate and severely malnourished groups; the time to no response on train-of-four stimulation was delayed only in severely malnourished groups (P < 0.001). The duration of action of the initial dose was shorter in the moderate and severely malnourished groups. The obese group had an earlier onset of action and a longer duration of action compared with patients of normal nutrition (P < 0.001). No significant difference in recovery time to a train-of-four ratio of 0.70 was observed between the malnourished and patients with normal nutrition. Malnutrition has a marked effect on vecuronium-induced neuromuscular blockade.

Adolescent↗

PubMed1998

A leg ulcer clinic: audit of the first three years.

This study documents the establishment in 1994 of a leg ulcer clinic in Hobart, Tasmania. Information relating to patient demographics, results of treatment and costs required to successfully operate the clinic were studied prospectively. Over a 40-month period, 345 patients attended the clinic on 1154 occasions. Ulcers were classified as venous (59%), arterial (19%), mixed venous and arterial disease (13%), pressure injury (3%), traumatic injury (2%) or rheumatoid disease (1%). A wide range of dressings and bandages were used during treatment. The clinic cost an average of A$42,683 per annum to run. This information has the potential to be of use as a guide for any organisation planning to open a similar clinic.

Adult↗

PubMed1998

Picroliv, the iridoid glycoside fraction of Picrorhiza kurroa, selectively augments human T cell response to mycobacterial protein antigens.

Mycobacterial and other intracellular parasitic diseases are characterised by a deficiency in antigen specific host T cell responses. We have studied the effect of Picroliv, a standardised fraction of root and rhizome of Picrorhiza kurroa, on proliferative T cell response to the mycobacterial 'Purified Protein Derivative (PPD)' antigen in subjects infected with or exposed to mycobacteria (tuberculoid leprosy patients and endemic normals). Coculture of their peripheral blood mononuclear cells with the optimal concentration of Picroliv (0.5 microgram/ml) significantly enhanced the proliferative response to 1/10 optimal PPD dose, as determined by [3H] thymidine incorporation, in the group of 'low' responders. The response to PPD of cells from 'high responders' and to PHA (phytohaëmagglutinin, a non-specific T cell mitogen) remained unaffected by Picroliv which did also not induce cell proliferation on its own. The selective, antigen specific augmentation of human T cell response suggests that Picroliv could be useful as an adjunct to chemotherapy or as a short term prophylactic agent.

Antigens, Bacterial↗

PubMed1998

Matrix metalloproteinases and abdominal aortic aneurysms: a potential therapeutic target.

Abdominal aortic aneurysm (AAA) is a leading cause of death in the United States, and there is no effective treatment in the early course of disease. Therapy to retard or reverse aneurysmal growth requires an understanding of the underlying vascular pathology. Recent research has indicated that enzymatic degradation of structural matrix proteins plays a large role in the formation of AAAs. Specifically, many studies have implicated a family of matrix degrading enzymes, known as matrix metalloproteinases (MMPs), as vital factors in the disease. Although AAA was once thought to be purely secondary to atherosclerosis, investigators have demonstrated various differences between the diseases in both levels and distribution of MMPs, suggesting independent mechanisms. Experimental models have shown that inhibition of these proteinases may slow aortic wall matrix breakdown. The purpose of this article is to review the current literature regarding the role of individual MMPs in AAA, including their complex regulatory mechanisms and possible cellular sources, the importance of MMPs as a potential therapeutic target in the prevention and treatment of AAA, and their inhibition using novel pharmacologic interventions in animal models.

Aortic Aneurysm, Abdominal↗

PubMed1998

Serology and immunoglobulin profile in rheumatoid arthritis.

One hundred and twenty cases of clinically diagnosed rheumatoid arthritis, 80 non-rheumatoid cases suffering from various other diseases and 40 healthy individuals were investigated for the presence of rheumatoid factor, quantitation of serum immunoglobulin, demonstration of ANA and LE cell phenomenon. Microlatex agglutination test of serum for rheumatoid factor showed 56.6% positivity in rheumatoid group and 3.7% positivity in non-rheumatoid group. All three serum immunoglobulins (IgG, IgM, IgA) were raised in serum in significant titre in cases of rheumatoid arthritis, whereas only IgA lever was elevated in the group of non-rheumatoid diseases. ANA and LE cell phenomenon were observed in 11.7% and 4.4% cases of rheumatoid arthritis who had severe underlying disease. In non-rheumatoid group, only one of 6 cases of systemic lupus erythematosus showed rheumatoid factor and that too in an insignificant titre (less than 1:20). Synovium and synovial fluid contained plenty of plasma cells and lymphocytes. It has been observed that RF appears first in synovial fluid and it may take several months to a year to attain detectable level in serum.

Adolescent↗

PubMed1998

Fluchloralin is cytotoxic and genotoxic and induces apoptosis in mammalian cells.

The genotoxic and cytotoxic effects of a widely used herbicide, fluchloralin, were assessed using cultured mammalian cells. Treatment of cells for 8-12 hr with fluchloralin resulted in a significant increase in the frequency of metaphase cells with chromosomal damage. At higher concentrations, the herbicide also induced an increase in the frequency of sister chromatid exchange. A 50% loss in viability was observed when cells were exposed to the herbicide for 72 hr. To understand the mechanism of cell death caused by fluchloralin, its effect on DNA synthesis and its ability to induce apoptosis were investigated. Even short (6 hr) treatment of cells with fluchloralin resulted in a 30-50% inhibition of DNA synthesis. Agarose gel electrophoresis of DNA from herbicide-treated cells and cytochemical staining indicate the induction of apoptosis by fluchloralin.

Aniline Compounds↗

PubMed1998

Hepatoprotective effect of a protein isolated from Cajanus indicus (Spreng) on carbon tetrachloride induced hepatotoxicity in mice.

Treatment with hepatotoxin namely carbon tetrachloride (CCl4) (0.1 ml/100 g of body weight; twice a week) induced acute hepatic necrosis in Swiss albino mice (male; body weight 30 g +/- 2), with significant alteration in the activities of glutamic oxaloacetic transaminase (GOT); glutamic pyruvic transaminase (GPT); alkaline phosphatase (AP) and serum bilirubin. Administration of a protein fraction isolated from the leaves of C. indicus counteracted the action of CCl4 on transaminase, phosphatase showing hepatoprotection. Daily treatment with a purified protein fraction 'X' from the above plant (0.5 mouse ml i.p; 50-60 micrograms/ml) for a period of 7, 14, 21 days respectively showed decreased activities of serum transaminases alkaline phosphatase and decreased levels of serum bilirubin. These findings were further confirmed by histopathological study of liver.

Animals↗

PubMed1997

Magnetic bead enzyme-linked immunosorbent assay (ELISA) detects antigen-specific binding by phage-displayed scFv antibodies that are not detected with conventional ELISA.

An efficient means for the detection of antigen-specific binding by phage-displayed antibodies would facilitate the selection of such phage, especially from libraries with large repertoires of V-genes. We report the development and characterization of a magnetic bead phage ELISA which detects antigen binding phage which could not be detected by conventional ELISA. We were attempting to select phage binding to the oncodevelopmental antigen, heat-stable alkaline phosphatase (HSAP). Although there was an obvious enrichment in the phage titers after successive rounds of selection, we were unable to detect antigen-binding phage by ELISA on a plastic surface. However, ELISA with a suspension of superparamagnetic particles covalently conjugated to HSAP effectively identified antigen-binding phage after the fourth round of selection. This method could also detect antigen-specific binding of individual phage clones. Some of the phage clones bound to either amino- or carboxy-terminal-conjugated HSAP, perhaps reflecting the differences in the exposed epitopes. It is suggested that a sensitive method such as magnetic bead phage ELISA be tried before declaring a phage selection as unsuccessful or concluding that a phage clone does not bind antigen.

Alkaline Phosphatase↗

PubMed1997

Neuropeptide Y receptor genes mapped in human and mouse: receptors with high affinity for pancreatic polypeptide are not clustered with receptors specific for neuropeptide Y and peptide YY.

Ppyr1, Npy5r, and Npy6r, the genes encoding mouse type 4, type 5, and type 6 members of the neuropeptide Y receptor family, have been mapped by interspecific backcross analysis to conserved linkage groups on mouse Chr 14, Chr 8, and Chr 18, respectively. The human genes, PPYR1 and NPY5R, have been localized to chromosomes 10q and 4q, respectively, by analysis of a panel of rodent-human somatic cell hybrids and yeast artificial chromosomes. These studies complete the mapping of the cloned NPY receptor subtypes in human and mouse and, together with previous studies, establish that the genes encoding receptors with high affinity for pancreatic polypeptide are not clustered with the genes encoding receptors specific for neuropeptide Y and peptide YY. The physical association of these receptor genes correlates with ligand-binding properties, rather than sequence identity, and suggests a complex evolutionary relationship.

Animals↗

PubMed1997

Comparison of caspase activation and subcellular localization in HL-60 and K562 cells undergoing etoposide-induced apoptosis.

Previous studies have shown that K562 chronic myelogenous leukemia cells are resistant to induction of apoptosis by a variety of agents, including the topoisomerase II (topo II) poison etoposide, when examined 4 to 24 hours after treatment with an initiating stimulus. In the present study, the responses of K562 cells and apoptosis-proficient HL-60 acute myelomonocytic leukemia cells to etoposide were compared, with particular emphasis on determining the long-term fate of the cells. When cells were treated with varying concentrations of etoposide for 1 hour and subsequently plated in soft agar, the two cell lines displayed similar sensitivities, with a 90% reduction in colony formation at 5 to 10 mu mol/L etoposide. After treatment with 17 mu mol/L etoposide for 1 hour, cleavage of the caspase substrate poly(ADP-ribose) polymerase (PARP), DNA fragmentation, and apoptotic morphological changes were evident in HL-60 cells in less than 6 hours. After the same treatment, K562 cells arrested in G2 phase of the cell cycle but otherwise appeared normal for 3 to 4 days before developing similar apoptotic changes. When the etoposide dose was increased to 68 mu mol/L, apoptotic changes were evident in HL-60 cells after 2 to 3 hours, whereas the same changes were observed in K562 cells after 24 to 48 hours. This delay in the development of apoptotic changes in K562 cells was accompanied by delayed release of cytochrome c to the cytosol and delayed appearance of peptidase activity that cleaved the fluorogenic substrates Asp-Glu-Val-Asp-aminotrifluoromethylcoumarin (DEVD-AFC) and Val-Glu-Ile-Asp-aminomethylcoumarin (VEID-AMC) as well as an altered spectrum of active caspases that were affinity labeled with N-(Nalpha-benzyloxycarbonylglutamyl-Nepsilon-biotin yllysyl) aspartic acid [(2,6-dimethylbenzoyl)oxy]methyl ketone [z-EK(bio)D-aomk]. On the other hand, the activation of caspase-3 under cell-free conditions occurred with indistinguishable kinetics in cytosol prepared from the two cell lines. Collectively, these results suggest that a delay in the signaling cascade upstream of cytochrome c release and caspase activation leads to a long latent period before the active phase of apoptosis is initiated in etoposide-treated K562 cells. Once the active phase of apoptosis is initiated, the spectrum and subcellular distribution of active caspase species differ between HL-60 and K562 cells, but a similar proportion of cells are ultimately killed in both cell lines.

Antineoplastic Agents, Phytogenic↗

PubMed1997

Loss of heterozygosity of a locus on 17p13.3, independent of p53, is associated with higher grades of astrocytic tumours.

Amongst the human astrocytic tumours, the commonest of primary brain tumours, the clinical outcome of astrocytoma (AS) is significantly better than anaplastic astrocytoma (AA) and glioblastoma multiforme (GBM). Often, low grade tumours can progress to or recur with a more malignant phenotype. Recent loss of heterozygosity (LOH) reports suspect the involvement of a tumour suppressor gene, different from p53, in the 17p13.3 region of the human chromosome. However, the effect of LOH of 17p13.3 region on tumour histology at presentation and prognosis is as yet undefined. As a first step to define the role of this putative oncogene in astrocytic tumour progression, we correlated the LOH of a locus, D17S379, in 17p13.3 region and the p53 locus in 17p13.1 region with the histopathology of astrocytic tumours by PCR based microsatellite and restriction fragment length polymorphism of DNA extracted from microdissected paraffin sections of 45 astrocytic tumours of different histopathological grades. LOH of D17S379 was significantly associated (P=0.02) with AA and GBM (high grade malignancy), while no such preferential association was found with LOH of p53. There were no mutations in the exons 5 to 9 of p53 gene in the five tumours with LOH of D17S379 but not of p53 region. In a case of AA with a heterogenous microscopic appearance, heterozygosity of D17S379 was lost only in the area with a more malignant histology while both areas had no LOH or mutation of p53. A locus at the 17p13.3 region, independent of the p53 locus, is involved in a large subset of astrocytic tumours during transformation into a more malignant phenotype, and thus may be a link in the chain of genetic events occurring in astrocytic tumour progression.

Astrocytoma↗

PubMed1997

Molecular cloning of a novel human gene encoding a 63-kDa protein and its sublocalization within the 11q13 locus.

A human cDNA previously isolated by virtue of its ability to complement partially the ultraviolet sensitivity of a xeroderma pigmentosum cell line was further characterized. The transcription unit is expressed as a single 4.0-kb mRNA that encodes a novel 63-kDa cytoplasmic protein, possibly initiating from an internal AUG codon. The gene encoding this protein, named UVRAG, has been extremely well conserved during evolution, implying an important role for this gene product in cell metabolism. The transcribed mRNA is constitutively expressed in a wide variety of human tissues. The protein encoded by this gene is predicted to contain a coiled-coil structure and is likely to be metabolically unstable based on the occurrence of a strong PEST domain. UVRAG was assigned to human chromosome 11 by Southern hybridization to a somatic cell hybrid panel. Fluorescence in situ hybridization coupled with PCR analysis of human/rodent somatic cell hybrids containing segments of human chromosome 11 has localized this gene to a subregion of 11q13 in between the D11S916 and the D11S906 loci. Importantly, this region has been shown to be amplified in a variety of human malignancies, including breast cancer.

Amino Acid Sequence↗

PubMed1997

Rectal response of cardiac origin in the cat: involvement of nitric oxide and acetylcholine.

Local application of nicotine over the surface of the left ventricle and also occlusion of the left anterior descending coronary artery in the lightly anaesthetised, open-chested, artificially ventilated cat resulted a biphasic rectal movement--initial relaxation followed by sustained contraction. However, distension of the atrial appendage did not evoke any change in rectal motility, indicating the non-involvement of atrial volume receptors in initiating this rectal response of cardiac origin. The relaxation phase of this response was not abolished by pretreatment with atropine or with phentolamine or propranolol but was abolished by the nitric oxide inhibitor, N(G)-nitro-L-arginine (LNNA), and this blockade of the relaxation phase by LNNA was reversed by L-arginine. The contraction phase, however, was abolished by atropine. From these observations it is clear that the relaxation phase of the rectal response to coronary occlusion or epicardial nicotine is mediated through neither cholinergic nor adrenergic pathways but through the release of nitric oxide whereas the contraction phase of such a cardio-rectal response is mediated through the release of the neurotransmitter, acetylcholine.

Acetylcholine↗

PubMed1997

Activation of multiple interleukin-1beta converting enzyme homologues in cytosol and nuclei of HL-60 cells during etoposide-induced apoptosis.

Recent genetic and biochemical studies have implicated cysteine-dependent aspartate-directed proteases (caspases) in the active phase of apoptosis. In the present study, three complementary techniques were utilized to follow caspase activation during the course of etoposide-induced apoptosis in HL-60 human leukemia cells. Immunoblotting revealed that levels of procaspase-2 did not change during etoposide-induced apoptosis, whereas levels of procaspase-3 diminished markedly 2-3 h after etoposide addition. At the same time, cytosolic peptidase activities that cleaved DEVD-aminotrifluoromethylcoumarin and VEID-aminomethylcoumarin increased 100- and 20-fold, respectively; but there was only a 1. 5-fold increase in YVAD-aminotrifluoromethylcoumarin cleavage activity. Affinity labeling with N-(Nalpha-benzyloxycarbonylglutamyl-Nepsilon-biotin yllysyl)aspartic acid [(2,6-dimethylbenzoyl)oxy]methyl ketone indicated that multiple active caspase species sequentially appeared in the cytosol during the first 6 h after the addition of etoposide. Analysis on one- and two-dimensional gels revealed that two species comigrated with caspase-6 and three comigrated with active caspase-3 species, suggesting that several splice or modification variants of these enzymes are active during apoptosis. Polypeptides that comigrate with the cytosolic caspases were also labeled in nuclei of apoptotic HL-60 cells. These results not only indicate that etoposide-induced apoptosis in HL-60 cells is accompanied by the selective activation of multiple caspases in cytosol and nuclei, but also suggest that other caspase precursors such as procaspase-2 are present but not activated during apoptosis.

Antineoplastic Agents, Phytogenic↗

PubMed1997

Oxidative stress induced by iron in Hydrilla verticillata (l.f.) Royle: response of antioxidants.

The effect of iron (FeCl3) on chlorophyll content, lipid peroxidation product, potassium ion leakage (a measure of damage to the permeability barrier), and antioxidants was studied in Hydrilla verticillata. The effect of iron-induced damage to the plant was compared with those of N-ethyl maleimide (NEM), a sulfhydryl reagent, and cumene hydroperoxide (CHP), an organic peroxide known to induce lipid peroxidation by free radical formation. The level of lipid peroxidation product was increased in the plants treated with Fe, CHP, and CHP + NEM but not with NEM alone. A significant increase in potassium ion leakage to the external solution was observed by the addition of Fe, CHP, and CHP + NEM, while this did not increase significantly in NEM-treated plants. When NEM and CHP were added simultaneously, the results were the same as those obtained with high iron concentrations, suggesting a combined effect of thiol depletion and lipid peroxidation by Fe ions. In addition, the results indicated loss of glutathione (GSH) and increased oxidized glutathione (GSSG) under Fe stress, indicative of oxidative stress. The oxidative stress may increase the production of free radicals and subsequently resulted in peroxidation of lipids. Further, addition of iron increased the activity of superoxide dismutase (SOD) which may be due to enhanced production of oxygen free radical and related tissue damage. The results suggest that iron-induced damage in plants can be ascribed to a direct metal action on thiols and by toxic oxygen species. An increase in lipid peroxidation product and K+ leakage are the primary responses of iron toxicity on membrane damage. However, the decrease in chlorophyll content is part of the overall expression of iron toxicity.

Antioxidants↗

PubMed1997

Proportion and pattern of dystrophin gene deletions in north Indian Duchenne and Becker muscular dystrophy patients.

Population-based variations in frequency and distribution of dystrophin gene deletions have been recognized in Duchenne/Becker (DMD/BMD) muscular dystrophy patients. In the present study, DNA samples from 121 unrelated DMD/BMD patients from North India were analyzed for deletional studies with multiplex PCR and Southern hybridization. A total of 88 (73%) patients showed intragenic deletions in the dystrophin gene. The observed proportion of gene deletions is relatively high, particularly compared with that of Asian counterparts. However, the distribution of breakpoints across the gene does not show significant variations.

Dystrophin↗

PubMed1997

Coronary artery calcium: alternate methods for accurate and reproducible quantitation.

RATIONALE AND OBJECTIVES: The aim of this study was to determine a more precise and accurate method of quantitating coronary artery calcium (CAC) detected with electron-beam computed tomography (CT) in patients with low CAC scores. MATERIALS AND METHODS: Two 40-section, 3-mm-collimation, electrocardiographically gated electron-beam CT examinations of the heart were performed in each patient. Fifty patients with average scores between 2 and 100, as determined with the conventional scoring algorithm, were selected. The modified conventional scoring algorithm was compared with two techniques: calculated calcium volume and approximated calcium mass. RESULTS: The percentage difference between scans ranged from 37.2% for the conventional scoring method to 28.2% and 28.4% for volume- and mass-based methods, respectively. Increasing lesion size thresholds does not improve quantitative precision and reduces accuracy in patients with small amounts of CAC. CONCLUSION: Quantification methods based on calcification volume or mass decrease score variation compared with the conventional scoring method, and increased size threshold does not improve accuracy.

Adult↗

PubMed1997

Fractionation of mycobacterial integral membrane proteins by continuous elution SDS-PAGE reveals the immunodominance of low molecular weight subunits for human T cells.

Integral membrane proteins (IMP) represent a serologically distinct class of mycobacterial antigens which are potent stimulators of human T cells (Mehrotra et al., Clin Exp Immunol 1995; 102:626). The range of IMP from Mycobacterium fortuitum was resolved by continuous elution SDS-PAGE to recover 31 discrete fractions covering bands up to approximately 58 kD. The fractions, after removal of SDS, were subjected to human T cell proliferation assays for the identification of immunodominant molecule(s). A low molecular weight (<20kD) fraction was able to stimulate T cells from 11 out of 12 donors comprising mainly tuberculoid leprosy patients. The described protocol is well suited to situations where large quantities of antigenic protein mixtures must be processed in order to get the purified molecules/fractions in amounts required for immunoepidemiological studies.

Bacterial Proteins↗