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Biomedical subjects

G Kumar

Publications and source records attributed to G Kumar.

At least 55 records · Page 3Linked to original sources

Human peripheral mononuclear cell responses to UV damage are affected by radiation-induced changes in plasma.

To evaluate the effects of environmental or therapeutic stress adequately, it is important to study cells or tissues under conditions that simulate as closely as possible the in vivo environment. To determine whether the responses of irradiated cells are significantly affected by radiation-induced changes in plasma, human mononuclear cells were isolated from peripheral blood and cultured in their autologous plasma. Freshly isolated cells were irradiated in phosphate-buffered saline. The plasma was irradiated separately. Irradiation of the plasma suppressed mitogen-induced DNA synthesis in unirradiated cells. For cells that were UV-damaged and subsequently stimulated with mitogen, DNA synthesis was enhanced by irradiation of the plasma. Medium in which irradiated cells had previously been incubated enhanced DNA, synthesis in unirradiated cells that had been mitogen stimulated but did not affect the UV-induced shutoff of DNA synthesis in replicating cells or unscheduled DNA synthesis in irradiated cells.

Cell Division↗

Pharmacokinetic enhancement of inhibitors of the human immunodeficiency virus protease by coadministration with ritonavir.

Coadministration with the human immunodeficiency virus (HIV) protease inhibitor ritonavir was investigated as a method for enhancing the levels of other peptidomimetic HIV protease inhibitors in plasma. In rat and human liver microsomes, ritonavir potently inhibited the cytochrome P450 (CYP)-mediated metabolism of saquinavir, indinavir, nelfinavir, and VX-478. The structural features of ritonavir responsible for CYP binding and inhibition were examined. Coadministration of other protease inhibitors with ritonavir in rats and dogs produced elevated and sustained plasma drug levels 8 to 12 h after a single dose. Drug exposure in rats was elevated by 8- to 46-fold. A > 50-fold enhancement of the concentrations of saquinavir in plasma was observed in humans following a single codose of ritonavir (600 mg) and saquinavir (200 mg). These results indicate that ritonavir can favorably alter the pharmacokinetic profiles of other protease inhibitors. Combination regimens of ritonavir and other protease inhibitors may thus play a role in the treatment of HIV infection. Because of potentially substantial drug level increases, however, such combinations require further investigation to establish safe regimens for clinical use.

Animals↗

Porcine S-antigen: cDNA sequence and expression in retina, ciliary epithelium and iris.

cDNA clones encoding S-antigen (arrestin) were isolated from the expression library constructed from porcine retina and sequenced. The 1490 base pair fragment contained a 1215 base pair open reading frame. From the nucleotide sequence, an amino acid sequence consisting of 405 residues was deduced and a molecular mass of 45,102 daltons was calculated. In order to determine whether the S-antigen mRNA transcript was expressed in anterior eye tissues, mRNA from ciliary non-pigmented epithelial cells and pigmented epithelial cells and iris was analysed by the reverse transcription polymerase chain reaction (PCR) using primers taken from sequences flanking the coding and non-coding regions of retinal S-antigen. Sequence analysis of the expected 611 base pairs in the 5' region and 672 base pairs in the 3' region of DNA fragments indicated that an identical mRNA for S-antigen was expressed in the anterior tissues examined. To investigate the in situ expression of S-antigen mRNA, 35S-labeled sense and antisense RNA probes were synthesized from the cDNA to label frozen sections of retina, ciliary body and iris and the treated sections were examined by autoradiography. The antisense probe labeled the layer between retinal pigmented epithelium and the outer nuclear layer of the retina, ciliary epithelium, and iris epithelium. From the results of sequencing PCR products and in situ hybridization, we concluded that, in porcine eye, the mRNA for S-antigen is expressed not only in the retina but also in the anterior tissues such as the ciliary epithelium and iris epithelium.

Animals↗

Biological denitration of propylene glycol dinitrate by Bacillus sp. ATCC 51912.

In previous studies, bacterial cultures were isolated that had the ability to degrade the nitrate ester glyceryl trinitrate (i.e., nitroglycerin). The goal of the present study was to examine the ability of resting cells and cell-free extracts of the isolate Bacillus sp. ATCC 51912 to degrade the more recalcitrant nitrate ester propylene glycol dinitrate (PGDN). It was observed that the PGDN-denitrating activity was expressed during growth even when cells were cultured in the absence of nitrate esters. This indicates that nitrate esters are not required for expression of denitration activity. Using cell-free extracts, PGDN was observed to be sequentially denitrated to propylene glycol mononitrate (PGMN) and propylene glycol with the second denitration step proceeding more slowly than the first. Also it was observed that dialysis of the cell-free extracts did not affect denitration activity indicating that regenerable cofactors [e.g., NAD(P)H or ATP] are not required for denitration.

Bacillus↗

Problems in differentiating sexually from nonsexually abused adolescent psychiatric inpatients by self-reported anxiety, depression, internalization, and externalization.

To ascertain whether self-reported psychopathology differentiated sexually and nonsexually abused adolescents, the Beck Depression Inventory, Beck Anxiety Inventory, and the Achenbach Youth Self-Report were administered to 111 psychiatric inpatients between 13 and 17 years of age who were diagnosed with various psychiatric disorders. Data about 14 background and clinical characteristics that were purported to be associated with sexual abuse were also collected. Forty (67%) of the 60 girls reported sexually abusive experiences, whereas six (12%) of the 51 boys reported such experiences. None of the scales were correlated with sexual abuse in either sex, and a history of physical abuse was the only characteristic that was significantly correlated with sexual abuse for both sexes. Furthermore, none of the scales was correlated with identity of sexual abuser, age of first abuse, age of last abuse, number of abuses, days of abuse, penile insertion, and the reporting of the abuse to the authorities in the sexually abused girls.

Adolescent↗

Zinc deficiency affects cell cycle and deoxythymidine kinase gene expression in HUT-78 cells.

Although zinc is known to be involved in cell proliferation and DNA synthesis, the mechanism by which zinc may regulate these processes is not understood. We have studied the role of zinc on cell proliferation and gene expression of a DNA synthesizing enzyme, deoxythymidine kinase (TK), in a T helper human malignant lymphoblastoid cell line (HUT-78). In zinc-deficient and zinc-sufficient media, the cell doubling time (mean +/- SD) of HUT-78 was 59 +/- 8 hours and 32.6 +/- 6 hours, respectively. The effect of zinc was T cell specific, inasmuch as the cell growth of another T malignant lymphoblastoid cell line, MOLT-3 (immature T cells), was not affected by zinc deficiency. Iron, copper, or manganese did not completely correct the cell growth of zinc-deficient HUT-78 cells. TK activity and the relative accumulation of TK-mRNA were significantly decreased in zinc-deficient cells during the G1 phase of cell cycle in comparison with zinc-sufficient cells. Nuclear run-on experiments and actinomycin-D studies showed that the transcription of TK-mRNA was affected adversely by zinc deficiency. Cell cycle studies showed that more zinc-deficient cells remained in S phase and did not undergo mitosis in comparison with zinc-sufficient cells. In conclusion, our data show that zinc is a T cell-specific growth factor and that a decreased gene expression of DNA-synthesizing enzyme TK in zinc-deficient HUT-78 cells in G1 phase affected adversely the DNA synthesis in S phase and delayed cell cycle.

Cell Cycle↗

The cerebellum-enriched form of nuclear factor I is functionally different from ubiquitous nuclear factor I in glial-specific promoter regulation.

Nuclear factor I (NFI) binding sites are present in a wide range of brain-specific gene enhancer and promoter sequences and appear to play a role in establishing cell type-specific expression within the CNS. The precise mechanisms used by various members of the NFI family of proteins to confer brain-specific expression are unclear. We have addressed this issue by comparing the transactivating capabilities of two forms of NFI in directing gliotropic expression from two different JC virus (JCV) promoter configurations. The JCV is an opportunistic pathogen of humans that causes lytic destruction of the oligodendrocytes and thus demyelination in immunocompromised patients. Our results show that the cerebellum-enriched form of NFI (NFI-A1) transactivates two gliotropic JCV early promoters to a greater extent than the ubiquitous form of NFI (NFI-C1). Activation by NFI-A1 was dramatically greater in glial than in nonglial cells. These results suggest that NFI proteins direct brain-specific expression through combinatorial interactions with cell specific coactivators and/or transcription factors that recognize adjacent sites within brain specific promoters.

Animals↗

Use of mid-arm and chest circumferences to predict birth weight in rural north India.

STUDY OBJECTIVE: To determine the most appropriate surrogate indicator and its cut off point for identifying low birthweight babies in northern India. STUDY SETTING: A secondary level hospital at Ballabgarh. The patients were from nearby rural and urban areas and mostly belonged to lower and middle socioeconomic strata. PARTICIPANTS: These comprised 733 singleton newborns delivered in the hospital between April and December 1991. DESIGN: Birth weight, arm circumference, and chest circumference were measured in all the newborns. Different cut off points for each index were identified and their validity was tested. Based on the regression equations, a simple chart was drawn up and was used to predict weights for different arm and chest circumferences in the hospital and community settings. MAIN RESULTS: Cut off points for arm and chest circumferences of 8.5 cm and 29.5 cm respectively gave a sensitivity and specificity of around 80%. When the chart based on the regression equations was tested in both the hospital and the community, chest circumference was found to be the better of the two indicators. CONCLUSION: Chest circumference seems to be the most appropriate surrogate measure for birth weight. Cut off points of 29.5 cm and 27.5 cm seem to be satisfactory for predicting birth weight below 2500 g and 1800 g respectively. The birthweight prediction card using chest circumference was effective in predicting birth weight.

Birth Weight↗

Frontoethmoid mucocele: one-year follow-up after endoscopic frontoethmoidectomy.

Endoscopic sinus surgery for frontal sinus mucocele is gaining popularity. A case study, and the CT scan of a large frontal mucocele with 1-year follow up is presented showing the lining of the sinus cavity well healed and the frontal sinus ostium as patent. Endoscopic sinus surgery, therefore, is a potentially useful method for treating large frontal mucoceles.

Endoscopy↗

Hepatic mesenchymal hamartoma: a case report and radiological findings.

Primary tumours of the liver are uncommon in childhood. Of these, more than two-thirds are malignant. As such, benign hepatic tumours are often not considered in the differential diagnosis of a hepatic mass in childhood. We report a case of hepatic mesenchymal hamartoma, a rare benign tumour, in a 10-month-old infant. This tumour is characterised by an admixture of ductal structures within a copious loose connective tissue stroma. Only approximately 160 cases had been reported in the literature. Awareness of the ultrasound (U/S) and computed tomography (CT) features, although not diagnostic, is helpful in distinguishing it from the more common malignant tumours. A correct preoperative diagnosis is important as surgical excision is often curative.

Diagnosis, Differential↗

The membrane immunoglobulin receptor utilizes a Shc/Grb2/hSOS complex for activation of the mitogen-activated protein kinase cascade in a B-cell line.

Ligation of membrane immunoglobulin M (mIgM) receptor in the Ramos B-cell line induced tyrosine phosphorylation of several intracellular substrates, including the adaptor protein. Shc. Phosphorylated Shc could be seen to associate with Grb2 in a complex which included hSOS. Inasmuch as hSOS is involved in p21ras activation, we also demonstrated that mIgM ligation activated a Ras-dependent kinase cascade in which sequential activation of Raf-1 and MEK-1 culminates in the activation of p42 mitogen-activated protein (MAP) kinase (ERK-2). The tumour promoter and protein kinase C agonist, phorbol 12-myristate 13-acetate (PMA), also activated Raf-1, MEK-1, and MAP kinase in Ramos cells, but did not induce tyrosine phosphorylation of Shc or Shc/Grb2 association. Okadaic acid, another tumour promoter and serine/threonine phosphatase inhibitor, activated p42 MAP kinase without activating Raf-1 or MEK-1, suggesting the existence of a serine/threonine phosphatase which directly regulates MAP kinase activity.

Adaptor Proteins, Signal Transducing↗

Evidence for LFA-1/ICAM-1 dependent stimulation of protein tyrosine phosphorylation in human B lymphoid cell lines during homotypic adhesion.

JK32.1 and SKW6.4 are Epstein-Barr virus (EBV)-positive human B cell lines that undergo spontaneous, lymphocyte function-associated antigen 1 (LFA-1) dependent homotypic adhesion in culture. This process is associated with induction of tyrosine phosphoproteins of molecular mass 90, 106, and 120 kDa and could be reproduced when these cells were centrifugationally aggregated. Antibodies to the beta 2 (CD18) chain of LFA-1 interfered with induction of p120, p106, and p90 during cellular aggregation. Response induction was abrogated when cells were incubated with protein tyrosine kinase (PTK) inhibitors (erbstatin, genistein, and geldanomycin) or cytochalasin B prior to aggregation. An in vitro kinase assay did not reveal activation of focal adhesion kinase. Although the role of LFA-1-dependent tyrosine phosphorylation in B cells is uncertain, patients with the leukocyte adhesion defect (LAD) exhibit humoral abnormalities. Moreover, aggregation did not induce specific tyrosine phosphoproteins in an EBV-transformed B cell line from a LAD patient. These results suggest that an LFA-1-dependent PTK pathway may play an important role in human B cell function.

Antibodies, Monoclonal↗

Comparative in vitro cytotoxic effects of taxol and its major human metabolite 6 alpha-hydroxytaxol.

Taxol is metabolized by the liver microsomal cytochrome P450 enzyme system into its principal metabolite 6 alpha-hydroxytaxol (6HT). In the present in vitro studies 6HT was compared to taxol with respect to its effects on tubulin depolymerization, mitotic arrest, clonogenic survival and apoptosis in HL-60 cells. 6HT was generated by incubating taxol with human liver microsomes in a NADPH-generating system. HL-60 cells were incubated for 24 h with either taxol or 6HT, washed and placed in drug-free suspension or cultured for colony growth in agarose. For the suspension and colony culture growth of the cells, the IC50 concentrations of 6HT were 500 +/- 46 and 350 +/- 37 nM, while those of taxol were 3.2 +/- 0.2 and 2.8 +/- 0.5 nM, respectively. Immediately after a 24-h exposure of HL-60 cells to 50 nM taxol, electrophoresis of genomic DNA from HL-60 cells revealed an internucleosomal DNA fragmentation 'ladder'. In addition, 39% of the cells were arrested in mitosis and 16% showed the morphologic features of apoptosis. In contrast, an identical treatment with 6HT resulted in the mitotic arrest of only 2.8% of the cells, with 4.0% displaying apoptosis (P < 0.01); internucleosomal DNA fragmentation was not observed. 6HT was also significantly less effective than taxol in inhibiting the temperature-induced depolymerization of microtubules in a cell-free system. However, at equipotent concentrations, the effect of 6HT on tubulin depolymerization, mitotic arrest or apoptosis was similar to that of taxol. In addition, at concentrations of taxol or 6HT at or below their IC50, there was little tubulin depolymerization, mitotic arrest or apoptosis. The results presented here show that the biotransformation of taxol to 6HT substantially detoxifies taxol.

Apoptosis↗

Denitration of glycerol trinitrate by resting cells and cell extracts of Bacillus thuringiensis/cereus and Enterobacter agglomerans.

A number of microorganisms were selected from soil and sediment samples which were known to have been previously exposed to nitrate ester contaminants. The two most effective bacteria for transforming glycerol trinitrate (GTN) were identified as Bacillus thuringiensis/cereus and Enterobacter agglomerans. For both isolates, denitration activities were expressed constitutively and GTN was not required for induction. Dialysis of cell extracts from both isolates did not affect denitration, which indicates that dissociable and depletable cofactors are not required for denitration. With thin-layer chromatography and high-performance liquid chromatography, the denitration pathway for both isolates was shown to be a sequential denitration of GTN to glycerol dinitrate isomers, glycerol mononitrate isomers, and ultimately to glycerol. GTN was observed to be completely converted to glycerol during a long-term incubation of cell extracts.

Bacillus cereus↗

The JC virus minimal core promoter is glial cell specific in vivo.

The glial cell specificity of the human papovavirus JC (JCV), an etiologic agent for progressive multifocal leukoencephalopathy, is thought to be due to the presence of both positive and negative regulatory elements upstream of the TATA region within the JCV promoter. Here we report that the JCV minimal core promoter, containing only the TATA box and an 8-bp poly(T) region immediately upstream, is sufficient to initiate transcription of an attached gene in glial cells and functions as an autonomously active initiator. We further define the sequences required for this core promoter's glial cell specificity by appropriate substitution and point mutation analysis. Ectopic expression of Tst-1, a POU domain transcription factor that has been implicated in the regulation of oligodendrocyte development, leads to higher activation of the JCV minimal core promoter in Tst-1-deficient glial cells than in non-glial HeLa cells. These results suggest a requirement for a glial cell coactivator(s) for the optimum activation of the JCV minimal core promoter by Tst-1. A discrete affinity of Tst-1 for the JCV core promoter (Kd, 1.4 x 10(-8) M) is also shown to be optimal for its promoter strength. Mutations within the core promoter that maintain this affinity for Tst-1 show maintenance of promoter strength, whereas mutants carrying a change that results in an increased affinity for Tst-1 show reduced transcriptional activity. These results suggest that moderate affinity of Tst-1 for the JCV TATA region may allow the interaction of some glial cell-specific coactivator(s) along with the basal transcription machinery to direct glial cell-specific transcription from the JCV core promoter.

Animals↗

Use of the Beck Anxiety Inventory with adolescent psychiatric outpatients.

The Beck Anxiety Inventory was administered to 105 outpatients between 13 and 17 years old who were diagnosed with various types of psychiatric disorders. A principal factor analysis was performed, and two factors were found representing subjective and somatic symptoms of anxiety. The item compositions of these factors were comparable to those previously described for adolescent inpatients. The results are discussed as supporting the use of the inventory for evaluating self-reported anxiety in outpatient adolescents.

Adolescent↗

Psychosocial correlates of suicidal ideation in adolescent psychiatric inpatients.

The Beck Scale for Suicide Ideation (BSI) was administered to 121 inpatients between 12 and 17 years old who were diagnosed with mixed psychiatric disorders. Twelve background and clinical characteristics, previously found to be associated with adolescent suicidal ideation, were entered into a multiple regression to estimate the BSI scores, along with the Beck Anxiety Inventory, the revised Beck Depression Inventory (BDI), the Beck Hopelessness Scale (BHS), and the Youth Self-Report. The BHS and the BDI were the only two variables that contributed unique variance to the explanation of the BSI scores.

Adolescent↗