Biomedical subjects

V Kumar

Publications and source records attributed to V Kumar.

At least 145 records · Page 8Linked to original sources

PubMed2000

Anxiolytic activity of Indian Abies pindrow Royle leaves in rodents: an experimental study.

Putative anxiolytic activity of ethanolic extract of Indian A. pindrow Royle leaf was investigated in rats using various experimental paradigms of anxiety viz. open field exploratory behaviour, elevated plus maze (EPM) and elevated zero maze (EZM) tests. Pilot studies indicated that single dose administration of extract had little to no acute behavioural effects, hence the extract was administered orally at different dose levels once daily for three consecutive days, while lorazepam (LR) (0.5 mg/kg, i.p.) was administered acutely. Ethanolic extract of A. pindrow (AP) leaves (50 and 100 mg/kg, p.o.) showed significant anxiolytic effects on all the paradigms of anxiety. The results indicate that AP and LR induced a significant increase in open field ambulation and slight increase in rearings and activity in center, whereas grooming and faecal droppings remain unchanged. In EPM, significant augmentation of open arm entries, and time spent on open arms was noted in AP treated rats. In EZM test, significant increase in time spent on open arms and entries in open arms was observed, whereas slight increase in head dips and stretched attend postures was also observed. The AP extract showed consistent and significant anxiolytic activity in all the tests. The effects induced by ethanolic extract of AP were less marked than those of lorazepam were.

Animals↗

PubMed2000

Anxiolytic activity of Indian Hypericum perforatum Linn: an experimental study.

The putative anxiolytic activity of 50% ethanolic extract of Indian Hypericum perforatum (IHp) was investigated in rats using various experimental paradigms of anxiety viz. open field exploratory behaviour (OFB), elevated plus maze (EPM), elevated zero maze (EZM), novelty induced suppressed feeding latency (FL) and social interaction (SI) tests. Pilot studies indicated that single dose administration of IHp had little to no acute behavioural effects, hence the extract of IHp was administered orally at different dose levels once daily for three consecutive days, while lorazepam (LR) (0.5 mg/kg, i.p.) was administered acutely. IHp extract (100 and 200 mg/kg, p.o.) showed significant anxiolytic effects on all the paradigms of anxiety. The results indicate that IHp and LR induced a significant increase in open field ambulation and slight increase in rearings and activity in centre, whereas grooming and fecal droppings remain unchanged. In EPM, significant augmentation of open arm entries, open arm/closed arm entries ratio and time spent on open arms was noted in IHp treated rats. In EZM test, significant increase in time spent on open arms and entries in open arms were observed, whereas slight increase in head dips and stretched attend postures were also observed. IHp and LR significantly attenuated the novelty induced increase in feeding latency. IHp treated rats also showed significant increase in social interaction in the novel environment. The IHp extracts showed consistent and significant anxiolytic activity in all the tests. The effects induced by 50% ethanolic extract of IHp were less marked than those of lorazepam were.

Administration, Oral↗

PubMed2000

Hypericum perforatum: nature's mood stabilizer.

Hypericum perforatum (HP), better known as St. John's Wort, has been used clinically for centuries. Modern usage is still quite diverse and includes kidney and lung ailments, insomnia and depression. Standardised extracts of HP are widely used in the treatment of psychovegetative disorders and especially for mild forms of depression. Several bioactive constituents of this plant may play important role in its well-known antidepressant activity, which are discussed in the present article. Furthermore, emphasis is also given on its botany, chemistry, pharmacology and clinical efficacy.

Antidepressive Agents↗

PubMed2000

An entomological field evaluation of larval biology of sandfly in Kala-azar endemic focus of Bihar--exploration of larval control tool.

Knowing the exact breeding places inside the habitat is very important to plan the larval control strategy. Information regarding larval biology in relation to different seasons will be more useful to organize insecticide spray schedule at a particular month of maximum immature density to bring down the adult sandfly density. In the present study, maximum number of soil samples were found positive in the month of January and minimum in the month of September. Maximum positive soil samples were collected from cattle sheds, minimum in mixed dwellings and in case of human dwellings all soil samples were negative. Comparison of two methods for the isolation of immature stages showed that direct microscopic observation is superior to sugar flotation technique.

Animals↗

PubMed1999

Perforin gene defects in familial hemophagocytic lymphohistiocytosis.

Familial hemophagocytic lymphohistiocytosis (FHL) is a rare, rapidly fatal, autosomal recessive immune disorder characterized by uncontrolled activation of T cells and macrophages and overproduction of inflammatory cytokines. Linkage analyses indicate that FHL is genetically heterogeneous and linked to 9q21.3-22, 10q21-22, or another as yet undefined locus. Sequencing of the coding regions of the perforin gene of eight unrelated 10q21-22-linked FHL patients revealed homozygous nonsense mutations in four patients and missense mutations in the other four patients. Cultured lymphocytes from patients had defective cytotoxic activity, and immunostaining revealed little or no perforin in the granules. Thus, defects in perforin are responsible for 10q21-22-linked FHL. Perforin-based effector systems are, therefore, involved not only in the lysis of abnormal cells but also in the down-regulation of cellular immune activation.

Antigen-Presenting Cells↗

PubMed1999

Interpolymer complexation. I. Preparation and characterization of a polyvinyl acetate phthalate-polyvinylpyrrolidone (PVAP-PVP) complex.

Polyvinyl acetate phthalate (PVAP) and polyvinylpyrrolidone (PVP) readily reacted in ethanol and acidic aqueous solutions to produce an insoluble PVAP-PVP complex. The complex has a pK(a) of 3.8. It is practically insoluble in common organic solvents, but dissolves in dimethylsulfoxide, an alkali or ammonical solution, and a 4:1 (v/v) mixture of methylene chloride and methanol. The powder X-ray diffraction analysis revealed the complex to be an amorphous material. The Fourier-transform infrared spectrum of the complex exhibited characteristics carbonyl stretching vibrations at 1724 and 1657 cm(-1) due to phthalate and acetate moieties in PVAP and cyclic amide groups in PVP, respectively, and at 1632 cm(-1) (appeared as a shoulder) due to cyclic amide groups of PVP bound to PVAP. The proton and carbon-13 (solution and solid-state) nuclear magnetic resonance spectra of the complex showed peak profiles that were linear combinations of those of PVAP and PVP. No new peaks appeared and no change in chemical shifts was observed due to complexation. The spectral data suggest that the interaction between PVAP and PVP probably initially involves the formation of hydrogen bonds between carbonyl groups of PVP and carboxylic groups of PVAP at some point of the polymer chains, causing the hydrophilic parts of the two flexible polymer chains strongly hydrophobic. As a result, the two polymer chains coil up into a compact structure and, consequently, precipitate out from the solution as an insoluble complex.

Chemistry, Pharmaceutical↗

PubMed1999

Differentiation of NK1.1+, Ly49+ NK cells from flt3+ multipotent marrow progenitor cells.

To delineate factors involved in NK cell development, we established an in vitro system in which lineage marker (Lin)-, c-kit+, Sca2+ bone marrow cells differentiate into lytic NK1.1+ but Ly49- cells upon culture in IL-7, stem cell factor (SCF), and flt3 ligand (flt3L), followed by IL-15 alone. A comparison of the ability of IL-7, SCF, and flt3L to generate IL-15-responsive precursors suggested that NK progenitors express the receptor for flt3L. In support of this, when Lin-, c-kit+, flt3+ or Lin-, c-kit+, flt3- progenitors were utilized, 3-fold more NK cells arose from the flt3+ than from the flt3- progenitors. Furthermore, NK cells that arose from flt3- progenitors showed an immature NK1.1dim, CD2-, c-kit+ phenotype as compared with the more mature NK1.1bright, CD2+/-, c-kit- phenotype displayed by NK cells derived from flt3+ progenitors. Both progenitors, however, gave rise to NK cells that were Ly49 negative. To test the hypothesis that additional marrow-derived signals are necessary for Ly49 expression on developing NK cells, flt3+ progenitors were grown in IL-7, SCF, and flt3L followed by culture with IL-15 and a marrow-derived stromal cell line. Expression of Ly49 molecules, including those of which the MHC class I ligands were expressed on the stromal or progenitor cells, as well as others of which the known ligands were absent, was induced within 6-13 days. Thus, we have established an in vitro system in which Ly49 expression on developing NK cells can be analyzed and possibly experimentally manipulated.

Animals↗

PubMed1999

Tolerance and alloreactivity of the Ly49D subset of murine NK cells.

Class I-specific stimulatory and inhibitory receptors expressed by NK cell subsets contribute to the alloreactive potential of the self-tolerant murine NK cell repertoire. In this report, we have studied potential mechanisms of tolerance to the function of the positive signaling Ly49D receptor in mice that express one of its ligands, H2-Dd. Our results demonstrate that H2-Dd-expressing mice possess a large Ly49D+ subset of NK cells that is functionally capable of rejecting bone marrow cell (BMC) allografts in vivo and lysing allogeneic Con A lymphoblasts in vitro. Also, we show that the Ly49D receptor is responsible for the ability of H2b/d F1 hybrid mice to reject H2d/d parental BMC (hybrid resistance). Thus, deletion or anergy of Ly49D+ cells in H2-Dd+ hosts cannot explain self tolerance. Our functional studies revealed that coexpression of the Dd-specific Ly49A or Ly49G2 inhibitory receptors by Ly49D+ cells resulted in tolerance to Dd+ targets, while coexpression of Kb-specific inhibitory receptors Ly49C/I resulted in tolerance to Kb+ targets. Only in H2d/d cells did Ly49C/I dominantly inhibit Ly49D-Dd stimulation. This correlated with an increased mean fluorescence intensity of Ly49C expression, as well as an increased percentage of Ly49C+ cells in the Ly49D+A/G2- compartment. Therefore, we conclude that self tolerance of the Ly49D subset can be achieved through coexpression of a sufficient level of self-specific inhibitory receptors.

Animals↗

PubMed1999

The X gene of hepatitis B virus shows a high level stimulation of the Rous sarcoma virus long terminal repeat in the methylotropic yeast, Pichia pastoris.

In order to study the transactivational property of the X gene in the methylotropic yeast Pichia pastoris, a Rous sarcoma virus-chloramphenicol acetyltransferase (RSV-CAT) cassette was co-transformed and integrated into the host yeast strain as a reporter which showed an overwhelming CAT activity. Immunoprecipitation of the yeast cell extracts with an X-specific monoclonal antibody, however, showed a low level expression of the X gene. Therefore besides a trans-effect of the X protein, the enhanced reporter activity could be a manifestation of a cis-effect of the X gene sequences also. Therefore, unlike the transactivation studies with X gene in animal cells where limited functional activity is observed, P. pastoris appears to be an excellent system to study cis- and trans-aspects of gene regulation by the X gene.

Avian Sarcoma Viruses↗

PubMed1999

Analysis of carboxyl content in oxidized celluloses by solid-state 13C CP/MAS NMR spectroscopy.

A noninvasive method to determine the carboxyl content in oxidized celluloses, using solid-state carbon-13 cross-polarization-magic angle spinning nuclear magnetic resonance (13C CP/MAS NMR) spectroscopy, has been developed. Standard samples containing 0, 4, 8, 12, 16, and 20% carboxyl content were prepared by mixing appropriate amounts of powdered cellulose, a non-oxidized cellulose standard prepared from cotton linter by ball-milling for 96 h, and a commercial oxidized cellulose sample that had a 20% carboxyl content. Standard curves were constructed by plotting the percent carboxyl content against the peak area at 171 ppm, normalized with (i) a peak area at 104 ppm and (ii) the sum of peak areas at 171, 62, and 65 ppm. The regression analysis of the curves yielded a linear relationship with correlation coefficient (R2) values of 0.9868 and 0.9863, respectively. To validate the methods, five new samples of oxidized cellulose were prepared and analyzed. The values obtained were comparable to those determined by the calcium acetate method described in the United States Pharmacopoeia (USP), indicating that the solid-state 13C CP/MAS NMR can be used to analyze carboxyl content in oxidized celluloses.

Acetates↗

PubMed1999

Cutting edge: expression of functional CD94/NKG2A inhibitory receptors on fetal NK1.1+Ly-49- cells: a possible mechanism of tolerance during NK cell development.

Fetal liver- and thymus-derived NK1.1+ cells do not express known Ly-49 receptors. Despite the absence of Ly-49 inhibitory receptors, fetal and neonatal NK1.1+Ly-49- cells can distinguish between class Ihigh and class Ilow target cells, suggesting the existence of other class I-specific inhibitory receptors. We demonstrate that fetal NK1. 1+Ly-49- cell lysates contain CD94 protein and that a significant proportion of fetal NK cells are bound by Qa1b tetramers. Fetal and adult NK cells efficiently lyse lymphoblasts from Kb-/-Db-/- mice. Qa1b-specific peptides Qdm and HLA-CW4 leader peptide specifically inhibited the lysis of these blasts by adult and fetal NK cells. Qdm peptide also inhibited the lysis of Qa1b-transfected human 721.221 cells by fetal NK cells. Taken together, these results suggest that the CD94/NKG2A receptor complex is the major known inhibitory receptor for class I (Qa1b) molecules on developing fetal NK cells.

Aging↗

PubMed1999

Activation of p38 mitogen-activated protein kinase by PYK2/related adhesion focal tyrosine kinase-dependent mechanism.

The stress-activated p38 mitogen-activated protein kinase (p38 MAPK), a member of the subgroup of mammalian kinases, appears to play an important role in regulating inflammatory responses, including cytokine secretion and apoptosis. The upstream mediators that link extracellular signals with the p38 MAPK signaling pathway are currently unknown. Here we demonstrate that pp125 focal adhesion kinase-related tyrosine kinase RAFTK (also known as PYK2, CADTK) is activated specifically by methylmethane sulfonate (MMS) and hyperosmolarity but not by ultraviolet radiation, ionizing radiation, or cis-platinum. Overexpression of RAFTK leads to the activation of p38 MAPK. Furthermore, overexpression of a dominant-negative mutant of RAFTK (RAFTK K-M) inhibits MMS-induced p38 MAPK activation. MKK3 and MKK6 are known potential constituents of p38 MAPK signaling pathway, whereas SEK1 and MEK1 are upstream activators of SAPK/JNK and ERK pathways, respectively. We observe that the dominant-negative mutant of MKK3 but not of MKK6, SEK1, or MEK1 inhibits RAFTK-induced p38 MAPK activity. Furthermore, the results demonstrate that treatment of cells with 1, 2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, tetra(acetoxymethyl)-ester, a membrane-permeable calcium chelator, inhibits MMS-induced activation of RAFTK and p38 MAPK. Taken together, these findings indicate that RAFTK represents a stress-sensitive mediator of the p38 MAPK signaling pathway in response to certain cytotoxic agents.

Animals↗

PubMed1999

Characterization of inhibitory and stimulatory forms of the murine natural killer cell receptor 2B4.

The receptor 2B4 belongs to the Ig superfamily and is found on the surface of all murine natural killer (NK) cells as well as T cells displaying non-MHC-restricted cytotoxicity. Previous studies have suggested that 2B4 is an activating molecule because cross-linking of this receptor results in increased cytotoxicity and gamma-interferon secretion as well as granule exocytosis. However, it was recently shown that the gene for 2B4 encodes two different products that arise by alternative splicing. These gene products differ solely in their cytoplasmic domains. One form has a cytoplasmic tail of 150 amino acids (2B4L) and the other has a tail of 93 amino acids (2B4S). To determine the function of each receptor, cDNAs for 2B4S and 2B4L were transfected into the rat NK cell line RNK-16. Interestingly, the two forms of 2B4 had opposing functions. 2B4S was able to mediate redirected lysis of P815 tumor targets, suggesting that this form represents an activating receptor. However, 2B4L expression led to an inhibition of redirected lysis of P815 targets when the mAb 3.2.3 (specific for rat NKRP1) was used. In addition, 2B4L constitutively inhibits lysis of YAC-1 tumor targets. 2B4L is a tyrosine phosphoprotein, and removal of domains containing these residues abrogates its inhibitory function. Like other inhibitory receptors, 2B4L associates with the tyrosine phosphatase SHP-2. Thus, 2B4L is an inhibitory receptor belonging to the Ig superfamily.

Animals↗

PubMed1999

Bcl-xL blocks activation of related adhesion focal tyrosine kinase/proline-rich tyrosine kinase 2 and stress-activated protein kinase/c-Jun N-terminal protein kinase in the cellular response to methylmethane sulfonate.

The stress-activated protein kinase/c-Jun N-terminal protein kinase (JNK) is induced in response to ionizing radiation and other DNA-damaging agents. Recent studies indicate that activation of JNK is necessary for induction of apoptosis in response to diverse agents. Here we demonstrate that methylmethane sulfonate (MMS)-induced activation of JNK is inhibited by overexpression of the anti-apoptotic protein Bcl-xL, but not by caspase inhibitors CrmA and p35. By contrast, UV-induced JNK activity is insensitive to Bcl-xL. The results demonstrate that treatment with MMS is associated with an increase in tyrosine phosphorylation of related adhesion focal tyrosine kinase (RAFTK)/proline-rich tyrosine kinase 2 (PYK2), an upstream effector of JNK and that this phosphorylation is inhibited by overexpression of Bcl-xL. Furthermore, overexpression of a dominant-negative mutant of RAFTK (RAFTK K-M) inhibits MMS-induced JNK activation. The results indicate that inhibition of RAFTK phosphorylation by MMS in Bcl-xL cells is attributed to an increase in tyrosine phosphatase activity in these cells. Hence, treatment of Bcl-xL cells with sodium vanadate, a tyrosine phosphatase inhibitor, restores MMS-induced activation of RAFTK and JNK. These findings indicate that RAFTK-dependent induction of JNK in response to MMS is sensitive to Bcl-xL, but not to CrmA and p35, by a mechanism that inhibits tyrosine phosphorylation and thereby activation of RAFTK. Taken together, these findings support a novel role for Bcl-xL that is independent of the caspase cascade.

Calcium-Calmodulin-Dependent Protein Kinases↗

PubMed1999

Biological parameters of Trichostrongylus colubriformis in Meriones unguiculatus.

In a series of three experiments, 64 jirds, Meriones unguiculatus, were infected with 700 infective larvae of Trichostrongylus colubriformis for the study of several biological parameters of this laboratory host-parasite model. In jirds the third stage larvae of T. colubriformis were shown to reach the fourth larval stage by the Day 6 post-infection (PI). By Day 10 PI, all the worms harvested had reached the immature adult stage. All immature adult stages of T. colubriformis developed to sexually mature adult stage by Day 12 PI when the female worms showed developing eggs in their uteri. Developed eggs were observed in the uteri of females on Days 13 and 14 PI. The first eggs of T. colubriformis appeared in the faeces of jirds on Day 13 PI. The peaks of egg production were recorded between Day 21 and Day 31 PI. Immunosuppression of jirds infected with 700 L3, by administration of dexamethasone from Day 57 to Day 94 PI led to increased faecal egg count when compared to untreated controls. All the worms were located in the small intestine. The jirds of dexamethasone treated group harboured higher number of adult worms than those of the untreated group. The number of adult worms was significantly higher in the first part of the small intestine than in the three other similar parts of the small intestine. The coefficient of correlation between the faecal egg count and worm number on the day of necropsy of jirds ranged between r = 0.58 and r = 0.89. Patent infections in jirds were maintained till the end of experiment on Day 100 PI, indicating that in this host and unlike other laboratory hosts, T. colubriformis is responsible for long lasting infections similar to what happens in domestic ruminants. The results of the present study suggest that the jird is a suitable laboratory model to study various aspects of this host-parasite relationship.

Animals↗

PubMed1999

Positive recognition of MHC class I molecules by the Ly49D receptor of murine NK cells.

Members of the murine Ly49 family of receptors have been shown to inhibit and activate NK cell function. Subsets of Ly49-expressing NK cells mediate the rejection of bone marrow cell allografts and the lysis of allogeneic lymphoblasts. In this report we have studied Ly49-mediated positive and negative signaling in an in vitro cytotoxicity assay using sorted NK cell subsets as effectors and a panel of 51Cr-labeled Con A lymphoblasts as targets in the presence or the absence of Abs to Ly49 and/or class I molecules. Our results demonstrate that the activating receptor Ly49D delivers stimulatory signals for target cell lysis upon interacting with H2-Dd, Dr, and Dsp2, but not H2b or H2k class I Ags. On the other hand, the inhibitory receptor Ly49G2 delivers negative signals for target cell lysis upon interacting with Dd, Dr, and H2k, but not H2b or Dsp2, class I Ags. Furthermore, Ly49-mediated negative signaling dominates Ly49D-mediated positive signaling. Thus, lysis of class I MHC-bearing targets by NK cells is not merely the consequence of the absence of an Ly49-mediated negative signal, but also requires positive recognition of class I molecules by certain Ly49 receptors. Activation of NK cells by nonself class I molecules was not predicted by the missing self hypothesis.

Animals↗

PubMed1999

Effect of compressional force on the crystallinity of directly compressible cellulose excipients.

The effect of compressional force on the crystallinity of low crystallinity cellulose (LCPC), microcrystalline celluloses (Avicel PH-101, PH-102 and PH-302 grades) and powdered cellulose (Solka Floc BW-100) has been investigated using an X-ray diffraction method. Microcrystalline and powdered celluloses showed an increase of about 10% in their crystallinities, compared to the values for the corresponding powders, at a compression pressure of 5-10 MPa. The increase in the crystallinity of LCPC was gradual and reached the maximum value of 5% at a compression pressure of 15 MPa. Further increase in compression pressure (to 77 MPa) had no effect on the crystallinity of LCPC, Avicel PH-101, Avicel PH-302 and Solka Floc BW-100. Avicel PH-102, on the other hand, showed a decrease in crystallinity at 15 MPa. Beyond 15 MPa, however, no statistically significant change in the crystallinity of the product was noted.

Cellulose↗

PubMed1999

The role of LY49 NK cell subsets in the regulation of murine cytomegalovirus infections.

The distributions and functions of NK cell subsets, as defined by the expression of Ly49 NK cell receptors, were examined in murine CMV (MCMV)-infected mice. MCMV induced a reduction in NK1.1+ cell number in the spleen and an increase in the peritoneal exudate cells. Within the splenic NK1.1+ population, proportional increases in Ly49A+ and Ly49G2+ cells but decreases in Ly49C+ and Ly49D+ cells were observed 3 days post-MCMV infection, but within the peritoneal NK1.1+ cell populations there were proportional decreases in Ly49A+ cells and increases in Ly49C+, Ly49D+, and Ly49G2+ cells. Lymphocytic choriomeningitis virus did not elicit a comparable NK cell subset distribution. Lymphokine-activated killer cells were sorted into different Ly49 NK cell subsets and adoptively transferred into C57BL/6 suckling mice. Regulation of MCMV synthesis in these suckling mice was shown to be an IFN-gamma-dependent, perforin- and Cmv-1-independent process, and each NK cell subset mediated anti-viral activity. In adult C57BL/6 mice, the control of MCMV in the spleen is mediated by a perforin-dependent mechanism, regulated in part by the Cmv-1 gene, which maps closely to the Ly49 family. In vivo depletions of either one or two of the Ly49 subsets in adult mice did not affect the ability of the residual NK cells to regulate MCMV synthesis. These data provide evidence of NK cell subset distribution and function in MCMV infection, but no individual subset was required for the Cmv-1-like regulation of MCMV synthesis.

Adoptive Transfer↗