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Biomedical subjects

J Canon

Publications and source records attributed to J Canon.

10 recordsLinked to original sources
PubMed2001

HIV type 1 Gag and nucleocapsid proteins: cytoskeletal localization and effects on cell motility.

Cell motility is likely to play a pivotal role in HIV infection by promoting the dissemination of infected cells. On the basis of observations indicating an interaction between HIV-1 Gag and target cell filamentous actin, we hypothesized that these interactions would promote cell motility of HIV-infected cells. Indeed, we have found that HIV-1 infection enhances the chemotactic response of macrophages. To specifically investigate the significance of the interactions between Gag and cellular actin, we transfected NIH 3T3 fibroblasts and HeLa cells with a construct that permits the expression of HIV-1 Gag in the absence of any other viral protein. Fractionation experiments showed that Gag was present in cytoskeletal fraction containing long actin filaments and in a high-speed postcytoskeletal fraction with short actin filaments. We have also localized HIV-1 Gag to the lamellipodia of chemoattractant-stimulated cells. Significantly, the motility of Gag-expressing cells was enhanced in chemotaxis assays. In vitro mutagenesis experiments showed that HIV-1 Gag binds filamentous actin through the nucleocapsid domain (NC). An NC-green fluorescent protein fusion had the same cellular distribution as the complete protein, and its expression increased cell motility. These data suggest that interactions between HIV-1 Gag and actin in infected cells enhance cell motility. Ultimately this enhanced motility of infected cells could promote the dissemination of virus into the brain and other tissues.

3T3 Cells↗
PubMed2000

Runt and Lozenge function in Drosophila development.

Runt and Lozenge (LZ) are members of the Runt domain family of transcriptional regulators and control a large number of developmental processes in Drosophila. Runt is a pair-rule gene, and is part of the network of genes that control pattern formation in the embryo. In the central nervous system, Runt function is necessary for the development of a subset of neurons. Runt is also a key regulator of sex determination, and directly controls Sex-lethal, a master gene that determines sex of the animal and controls dosage compensation. The LZ protein also participates in several key processes. LZ controls pre-patterning and cell-fate choices in the development of the visual system by regulating the expression of several fate-specifying transcription factors, and works in conjunction with general signaling pathways. LZ function is also required in hematopoiesis for the specification of a Drosophila blood cell lineage.

Animals↗
PubMed2000

Highly purified CD34+ cells isolated using magnetically activated cell selection provide rapid engraftment following high-dose chemotherapy in breast cancer patients.

The primary objective of this study was to evaluate the safety of infusion of CD34+ cells, selected using a clinical scale magnetically activated cell sorting device, assessed by time to hematological engraftment and incidence of adverse events. Secondary objectives included evaluation of device performance in terms of purity and recovery of the CD34+ cell product. Breast cancer patients suitable for transplantation received cyclophosphamide and filgrastim for mobilisation, followed by three leukaphereses. The products of the first two leukaphereses underwent CD34+ cell selection. The product of the third leukapheresis was cryopreserved unmanipulated. Following high-dose cyclophosphamide, thiotepa and carboplatin, selected CD34+ cells were infused. In 54 patients who received selected cells only, the median time to platelet recovery and neutrophil recovery was 11 days (range 5-51) and 9 days (range 5-51), respectively. There were no adverse events associated with infusion of selected cells. A total of 126 leukapheresis samples was available before and after selection for central CD34+ analysis. The median purity was 96.1% (27.4-99.4) and the median recovery was 52. 3% (15.2-146.3). These data show that cells selected using magnetically activated cell selection provide safe and rapid engraftment after high-dose therapy. Bone Marrow Transplantation (2000) 25, 243-249.

Adolescent↗
PubMed1998

Cells with high cyclophilin A content support replication of human immunodeficiency virus type 1 Gag mutants with decreased ability to incorporate cyclophilin A.

Gag polyprotein-mediated incorporation of cellular cyclophilin A (CyPA) into virions is essential for the formation of infectious human immunodeficiency virus type 1 (HIV-1) virions. Either a point mutation in Gag (P222A) or drugs which bind CyPA decrease virion incorporation of CyPA and interfere with HIV-1 replication. We have found that lymphoid cells varied greatly in their CyPA content and that cells with high CyPA content supported the replication of P222A HIV-1 Gag mutants. These experiments demonstrated that a higher cellular CyPA content of some cells was able to compensate for the decreased binding affinity of P222A mutant Gag for CyPA, allowing virus replication to occur.

Animals↗
PubMed1998

Cryoelectron microscopic examination of human immunodeficiency virus type 1 virions with mutations in the cyclophilin A binding loop.

The human immunodeficiency virus type 1 capsid protein contains a conserved P217X4PX2PX5P231 motif. Mutation at Pro-222 decreases virion incorporation of cyclophilin A, while mutation at Pro-231 abolishes infectivity. Although viral RNA incorporation and protease cleavage of the Gag precursor were not affected by these mutations, cryoelectron microscopy revealed a loss of virion maturation in P231A particles.

Binding Sites↗
PubMed1997

Towards interbreed IBD fine mapping of the mh locus: double-muscling in the Asturiana de los Valles breed involves the same locus as in the Belgian Blue cattle breed.

The Spanish "Asturiana" cattle breed is characterized by the segregation of a genetically determined muscular hypertrophy referred to as double-muscling or "culones". We demonstrate by linkage analysis that this muscular hypertrophy involves the mh locus previously shown to cause double-muscling in the Belgian Blue cattle breed, pointing towards locus homogeneity of this trait across both breeds. Moreover, using a twopoint and multipoint maximum likelihood approach, we show that flanking microsatellite markers are in linkage disequilibrium with the mh locus in both breeds albeit with different alleles. Finally, we discuss how allelic homogeneity across breeds might be exploited to achieve efficient genetic fine-mapping of the mh locus.

Animals↗
PubMed1996

Quantitative analysis of the endogenous reverse transcriptase reactions of HIV type 1 variants with decreased susceptibility to azidothymidine and nevirapine.

A large number of nucleoside analog and nonnucleoside inhibitors of HIV-1 reverse transcriptase (RT) have been developed for clinical use. Data confirm that resistant variants of HIV-1 rapidly emerge in response to the selective pressure of treatment with these agents. Detection of drug resistance generally involves detection of specific mutations in the viral genome or demonstrating a failure of the drug to suppress virus replication in culture. We have developed a PCR-based method to quantitatively examine HIV-1 DNA synthesis in vitro in endogenous reverse transcription reactions and tested it as a method to detect resistance to RT inhibitors. Under certain conditions, we were able to distinguish HIV strains with high-level resistance to azidothymidine triphosphate inhibition from sensitive strains. This method was quite useful as an assay to detect resistance to nevirapine, a nonnucleoside RT inhibitor; in reconstruction experiments, nevirapine-resistant virus was detectable when it represented 10 to 25% of the total amount of virus present in reaction mixtures. These data are examined in the light of current models of the mechanisms of action of nucleoside nonnucleoside RT inhibitors. This assay may be useful for detecting the emergence of drug-resistant HIV-1 variants during therapy.

Acquired Immunodeficiency Syndrome↗
PubMed1996

HIV-1 Gag protein associates with F-actin present in microfilaments.

Several studies have provided evidence that the cellular cytoskeleton may be involved in the assembly and budding of retroviruses. In fractionation studies of HIV-1-infected CEM cells, the majority of the unprocessed Gag polyprotein cofractionated with the cellular cytoskeleton. In vivo and in vitro analyses of this interaction indicated that the unprocessed Gag polyprotein is capable of association with polymerized actin (F-actin). Binding of Gag to F-actin may be involved in the assembly or budding of HIV-1.

Actin Cytoskeleton↗
PubMed1986

Preclinical evaluation of behavioral deficits following hypnotic drug treatment.

Most hypnotic medications result in performance decrements, or "hangover effects," the morning after bedtime ingestion. However, the severity of hangover effects with new compounds is difficult to predict prior to clinical trials. This paper describes a preclinical procedure that is predictive of human hangover effects following bedtime ingestion of hypnotic medication. Four cebus monkeys were shaped to discriminate between two levers such that rapid responses following the cue light maximized the probability of reinforcement. Subsequently, performance was evaluated one hr (immediate condition) and 8 3/4 hr (delayed condition) after the administration of flurazepam, triazolam, pentobarbital and placebo. All drugs caused dose-dependent performance decrements in both conditions. Significantly higher doses of triazolam were required to impair performance during the delayed condition as compared to the immediate condition, but no such differences occurred with either flurazepam or pentobarbital. These findings parallel human data which indicate that sleep-inducing doses of flurazepam and many barbiturates, but not triazolam, cause performance decrements the morning after bedtime ingestion.

Animals↗