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Biomedical subjects

R Fotedar

Publications and source records attributed to R Fotedar.

At least 37 records · Page 2Linked to original sources
PubMed1996

p21 contains independent binding sites for cyclin and cdk2: both sites are required to inhibit cdk2 kinase activity.

Cyclin dependent kinases regulate the progression of eukaryotic cells through the cell cycle. p21Cip1/Waf1/Sdi1 is an inhibitor of cdk-cyclin kinase activity, and has been shown to form complexes with cdk-cyclins and with PCNA, an accessory protein of DNA polymerase delta. The kinase inhibitory domain maps to the N-terminus (1-82) and contains the cdk2 binding site (28-82). We have generated a panel of deletion mutants of p21. A functional characterization of p21 mutants in the N-terminal domain reveals that cyclins bind to this domain independently of cdk2. Correlating with these results we find that p21 can associate with cyclin-cdk kinases in two functionally distinct forms, one in which the kinase activity is inhibited and the other in which the kinase is still active. The cdk2 and cyclin binding sites on p21 are both required to inhibit kinase activity. The second type of interaction, in which an active cyclin-cdk complex only interacts with p21 either via the cyclin or the cdk2 binding site but not through both, does not lead to inhibition of cyclin kinase activity. These results thus provide a basis for understanding the mechanism by which p21, and perhaps other cdk-cyclin kinase inhibitory proteins, suppress kinase activity.

Amino Acid Sequence↗
PubMed1996

Apoptosis and the cell cycle.

Apoptosis is a genetically controlled response by which eukaryotic cells undergo programmed cell death. This phenomenon plays a major role in developmental pathways (1), provides a homeostatic balance of cell populations, and is deregulated in many diseases including cancer. Control of cell number is determined by an intricate balance of cell death and cell proliferation. Accumulation of cells through suppression of death can contribute to cancer and to persistent viral infections, while excessive death can result in impaired development and in degenerative diseases. Identification of genes that control cell death, and understanding of the impact of apoptosis in both development and disease has advanced our knowledge of apoptosis in the past few years. There appears to be a linkage between apoptosis and cell cycle control mechanisms. Elucidating the mechanisms that link cell cycle control with apoptosis will be of key importance in understanding tumour progression and designing new models of effective tumour therapy.

Animals↗
PubMed1996

Changing pattern of Candida species in a bone marrow transplant patient.

A report on a patient with multiple myeloma, undergoing bone marrow transplant, who developed systemic fungal infection due to Candida krusei after bone marrow transplantation. The subsequent appearance of Ketoconazole resistant C. krusei is probably the first report of its kind from India. The significance of the findings is discussed.

Adult↗
PubMed1995

Cell cycle control of DNA replication.

The cell cycle is driven by the sequential activation of a family of cyclin-dependent kinases (cdk), which phosphorylate and activate proteins that execute events critical to cell cycle progression. In mammalian cells cdk2-cyclin A has a role in S phase. Many replication proteins are potential substrates for this cdk kinase, suggesting that initiation, elongation and checkpoint control of replication could all be regulated by cdk2. The association of PCNA, a replication protein, with cdk-cyclins during G-1 to S phase transition and with cdk-cyclin inhibitors, adds an interesting complexity to regulation of DNA replication.

Animals↗
PubMed1995

Changes in the subcellular localization of replication initiation proteins and cell cycle proteins during G1- to S-phase transition in mammalian cells.

DNA replication in eukaryotic cells is restricted to the S-phase of the cell cycle. In a cell-free replication model system, using SV40 origin-containing DNA, extracts from G1 cells are inefficient in supporting DNA replication. We have undertaken a detailed analysis of the subcellular localization of replication proteins and cell cycle regulators to determine when these proteins are present in the nucleus and therefore available for DNA replication. Cyclin A and cdk2 have been implicated in regulating DNA replication, and may be responsible for activating components of the DNA replication initiation complex on entry into S-phase. G1 cell extracts used for in vitro replication contain the replication proteins RPA (the eukaryotic single-stranded DNA binding protein) and DNA polymerase alpha as well as cdk2, but lack cyclin A. On localizing these components in G1 cells we find that both RPA and DNA polymerase alpha are present as nuclear proteins, while cdk2 is primarily cytoplasmic and there is no detectable cyclin A. An apparent change in the distribution of these proteins occurs as the cell enters S-phase. Cyclin A becomes abundant and both cyclin A and cdk2 become localized to the nucleus in S-phase. In contrast, the RPA-34 and RPA-70 subunits of RPA, which are already nuclear, undergo a transition from the uniform nuclear distribution observed during G1, and now display a distinct punctate nuclear pattern. The initiation of DNA replication therefore most likely occurs by modification and activation of these replication initiation proteins rather than by their recruitment to the nuclear compartment.

Base Sequence↗
PubMed1995

Activation-induced T-cell death is cell cycle dependent and regulated by cyclin B.

Developing thymocytes and some T-cell hybridomas undergo activation-dependent programmed cell death. Although recent studies have identified some critical regulators in programmed cell death, the role of cell cycle regulation in activation-induced cell death in T cells has not been addressed. We demonstrate that synchronized T-cell hybridomas, irrespective of the point in the cell cycle at which they are activated, stop cycling shortly after they reach G2/M. These cells exhibit the diagnostic characteristics of apoptotic cell death. Although p34cdc2 levels are not perturbed after activation of synchronously cycling T cells, cyclin B- and p34cdc2-associated histone H1 kinase activity is persistently elevated. This activation-dependent induction of H1 kinase activity in T cells is associated with a decrease in the phosphotyrosine content of p34cdc2. We also demonstrate that transient inappropriate coexpression of cyclin B with p34cdc2 induces DNA fragmentation in a heterologous cell type. Finally, in T cells, cyclin B-specific antisense oligonucleotides suppress activation-induced cell death but not cell death induced by exposure to dexamethasone. We therefore conclude that a persistent elevation of the level of cyclin B kinase is required for activation-induced programmed T-cell death.

Amino Acid Sequence↗
PubMed1993

p70 lupus autoantigen binds the enhancer of the T-cell receptor beta-chain gene.

The p70 (Ku) autoantigen has been described as a nonhistone nuclear protein recognized by antibodies from lupus patients. In our studies on the regulation of T-cell receptor (TCR) beta-chain gene expression we have identified the p70 lupus autoantigen as a DNA-binding protein that binds the enhancer of the TCR beta-chain gene. This enhancer is essential for expression of the TCR beta gene. The core TCR beta enhancer contains the E3 motif, which we show here is essential for enhancer activity. The protection of the E3 motif in T cells and the marked reduction in enhancer activity when the E3 motif is mutated underline its physiological importance in regulating beta enhancer activity. The p70 lupus autoantigen gene was identified by screening T-cell lambda gt11 libraries with an E3 probe. The gene encodes a protein which binds the E3 motif in a sequence-specific manner. The identification of a 70-kDa protein as a major E3-binding protein by UV crosslinking is consistent with the conclusion that the p70 lupus autoantigen binds the beta enhancer. Finally, we have shown that T-cell nuclear proteins which bind the E3 motif bear p70 (Ku) lupus autoantigenic determinants. Together these data suggest that the p70 autoantigen binds a critical motif in the beta enhancer and probably regulates TCR beta gene expression.

Animals↗
PubMed1993

Vector potential of the German cockroach in dissemination of Pseudomonas aeruginosa.

Survival time studies of Pseudomonas aeruginosa in cockroaches (Blattella germanica) were carried out under standard laboratory conditions. Cockroaches were fed with graded doses (10(2), 10(5) or 10(7) per insect) of P. aeruginosa. Depending on the excretion of P. aeruginosa the cockroaches were categorized as non-excretors, intermittent excretors or continuous excretors. When a dose of 10(2) P. aeruginosa was used all the insects were non-excretors but with doses of 10(5) or 10(7) per insect, P. aeruginosa multiplied in the gut of the cockroaches and was excreted for up to 114 days. The significance of these findings is discussed.

Animals↗
PubMed1992

Mapping of an inducible element in the T cell receptor V beta 2 promoter.

The murine V beta 2 promoter was analyzed for an element regulating phorbol ester inducibility of the TCR beta chain gene. In transient expression analysis of 5' nested deleted fragments of the V beta 2 promoter, the TPA-inducible element mapped between -85 and -42. The -85 to -62 oligo conferred 12-0-tetradecanoylphorbol-13-acetate (TPA) inducibility to the heterologous TPA-uninducible thymidine kinase promoter. The -85 to -62 region contained an AP-1 site (-85 to -72) and inverted repeat motif (-72 to -62). The AP-1 site required the 3' flanking inverted repeat region for conferring optimal inducibility. In vitro transcribed and translated jun/fos heterodimers bind to the V beta 2 AP-1 motif with a 16-fold lower affinity as compared to the collagenase AP-1 motif. This explains the inability of the V beta 2 AP-1 motif to confer optimal TPA inducibility by itself. The affinity of jun/fos heterodimers for the V beta 2 AP-1 motif was not increased by the presence in cis of the inverted repeat motif. The 3' flanking inverted repeat binds the ets transactivator but not jun/fos heterodimers. The demonstrated cooperativity between the AP-1 and the 3' flanking sequence to confer TPA inducibility can thus be explained by the individual contributions of jun/fos and ets transactivators.

Animals↗
PubMed1992

Cell cycle regulated phosphorylation of RPA-32 occurs within the replication initiation complex.

The transition from G1 to S phase of the cell cycle may be regulated by modification of proteins which are essential for initiating DNA replication. One of the first events during initiation is to unwind the origin DNA and this requires a single-stranded DNA binding protein. RPA, a highly conserved multi-subunit single-stranded DNA binding protein, was first identified as a cellular protein necessary for the initiation of SV40 DNA replication. The 32 kDa subunit of RPA has been shown to be phosphorylated at the start of S phase. Using SV40 replication as a model, we have reproduced in vitro the S phase-dependent phosphorylation of RPA-32 and show that it occurs specifically within the replication initiation complex. Phosphorylated RPA-32 is predominantly associated with DNA. Phosphorylation is not a pre-requisite for association with DNA, but occurs after RPA binds to single-stranded DNA formed at the origin during the initiation phase. The protein kinase(s) which phosphorylates RPA-32 is present at all stages of the cell cycle but RPA-32 does not bind to the SV40 origin or become phosphorylated in extracts from G1 cells. Therefore, the cell cycle-dependent phosphorylation of RPA-32 may be regulated by its binding to single-stranded origin DNA during replication initiation.

Animals↗
PubMed1992

The housefly (Musca domestica) as a carrier of pathogenic microorganisms in a hospital environment.

Houseflies have long been regarded as potential carriers of microorganisms. Since pathogenic microorganisms are widespread in the hospital environment, there is abundant opportunity for flies to become contaminated and, in turn, to contaminate the patient environment. In the present study, an attempt was made to isolate and identify pathogenic bacteria, fungi and parasites from the housefly Musca domestica collected in the surgical ward of the All India Institute of Medical Sciences Hospital and also in a remote residential area located 5 km from the hospital. A total of 113 flies were collected: 65 from a surgical ward (test) and 48 from a residential area for comparison. Ten genera of bacteria were isolated from the test group of flies compared with nine from the control group. In primary isolations, it was observed that the load of bacteria carried by the test group of flies was significantly more (P less than 0.001) than for the control flies. Pseudomonas aeruginosa, Enterococcus faecalis and viridans streptococci were isolated only from the test flies. The isolation rate of Staphylococcus aureus was significantly higher (P less than 0.001) in test houseflies than in the control houseflies. There was no significant difference in isolation of parasitic ova and cysts from test and control houseflies. Candida spp. were isolated in almost equal numbers from both groups of houseflies, yet none of these was Candida albicans. Houseflies therefore may act as vectors of potentially pathogenic bacteria in a hospital environment.

Animals↗
PubMed1992

Vector potential of hospital houseflies with special reference to Klebsiella species.

The vector potential of houseflies (Musca domestica) for Klebsiella spp. was investigated. Klebsiella spp. (mostly Klebsiella pneumoniae) were isolated from 36.7% of hospital flies and 28.1% of infected wounds of patients. Antibiograms of Klebsiella spp. showed that 82.0% of isolates from hospital flies and 96.3% from infected wounds were resistant to four or more commonly used antimicrobials. In contrast, from the control group, only 8.7% klebsiella isolates showed similar antimicrobial resistance pattern (P less than 0.001). Similar strains of Klebsiella spp. were encountered among patients and hospital houseflies. The results indicate that flies in the hospital environs are potential vectors of hospital resistant strains of Klebsiella spp.

Animals↗
PubMed1991

Nosocomial infections: cockroaches as possible vectors of drug-resistant Klebsiella.

The possibility that hospital cockroaches may act as vectors of drug-resistant Klebsiella spp. was investigated during Nov 1985 to April 1989, at the All India Institute of Medical Sciences (AIIMS) hospital. Klebsiella spp. (majority Klebsiella pneumoniae) were isolated from 28.3% of hospital cockroaches and 28.1% of infected wounds of patients. Most of Klebsiella isolates from patients (96.3%), and hospital cockroaches (85.9%) showed multiple drug resistance to four or more antimicrobials. Similar strains of Klebsiella spp. were encountered among patients and hospital cockroaches. These findings suggest that hospital cockroaches may act as vectors of drug-resistant Klebsiella spp. and may contribute to the epidemiology of nosocomial infections.

Animals↗
PubMed1991

Cockroaches (Blattella germanica) as carriers of microorganisms of medical importance in hospitals.

A study was conducted to isolate and identify microorganisms of medical importance from cockroaches (Blattella germanica) and to ascertain their vector potential in the epidemiology of nosocomial infections. Bacteria, fungi and parasites of medical importance were isolated and identified. Important bacterial pathogens responsible for wound infections, were further studies by antibiograms. One hundred and fifty-eight out of 159 (99.4%) cockroaches collected from hospital (test) and 113 out of 120 (94.2%) cockroaches collected from residential areas (control) were carrying medically important microorganisms (P less than 0.05). significantly higher (P less than 0.001) number of test cockroaches were carrying a higher bacterial load (1 x 10(4) and 1 x 10(5] as compared to control cockroaches. Multiple drug-resistant bacterial were isolated from test cockroaches. The diversity of drug-resistant bacterial species isolated from test cockroaches suggests their involvement in the transmission of drug-resistant bacteria. Various fungi and parasitic cysts of medical importance were also isolated from the test and control cockroaches, but the carriage rates were low. The findings suggest that cockroaches, in hospitals, can act as potential vectors of medically important bacteria/parasites/fungi.

Animals↗
PubMed1991

Human cutaneous myiasis due to mixed infestation in a drug addict.

Soft tissue infestation by maggots of flies (myiasis) is a well-recognized complication of neglected wounds. Mixed infestation by larvae of more than one species of fly is a rarity. We report, for the first time in India, one such case of cutaneous myiasis due to mixed infestation--in this instance by Chrysomya bezziana and Sarcophaga sp. in a drug addict.

Adult↗