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Biomedical subjects

D York

Publications and source records attributed to D York.

At least 55 records · Page 3Linked to original sources
PubMed1991

Pancreatic procolipase propeptide, enterostatin, specifically inhibits fat intake.

Pancreatic procolipase is activated by trypsin forming colipase, a cofactor for pancreatic lipase involved in intestinal fat digestion and a pentapeptide named enterostatin. Enterostatin with the sequence Val-Pro-Asp-Pro-Arg (VPDPR) was previously shown to decrease food intake in rats both after peripheral and central injection. In this work enterostatin has been shown to reduce specifically the consumption of a high-fat diet as opposed to a low-fat diet after central injection of Sprague-Dawley rats. After starvation for 18 hours the rats were given a free choice of a low-fat diet (5.2% fat by weight; 14.1% by energy) and a high-fat diet (17.8% fat by weight; 32.8% by energy) in separate containers. After injection of 200 ng of VPDPR into the lateral ventricle, the rats selectively decreased the intake of the high-fat diet by 45% (p less than 0.005), while the intake of the low-fat diet was unaffected compared to saline injection. VPDP after intracerebroventricular injection had totally lost the selective effect on the consumption of a high- fat and a low-fat diet. It is suggested that enterostatin formed during fat digestion from pancreatic procolipase may provide a feed-back signal for the intake of lipid.

Animals↗
PubMed1990

Recombinant gamma interferon in treatment of patients with atopic dermatitis and elevated IgE levels.

PURPOSE: Recombinant gamma interferon (rIFN-gamma) inhibits IgE synthesis in vitro by human peripheral blood mononuclear cells (PBMC). These data suggest a role for rIFN-gamma in the treatment of patients with severe atopic dermatitis (AD) and elevated IgE levels. The purpose of this study was to determine the effect of rIFN-gamma treatment on IgE production in patients with AD. PATIENTS AND METHODS: Twenty-two patients with chronic severe AD were treated with rIFN-gamma. In part I of the study, 14 patients were treated with daily subcutaneous injections at three successive dose levels (0.01 mg/m2, 0.05 mg/m2, and 0.1 mg/m2) for 5 days with 2 days off between each dose level. In part II, eight patients received rIFN-gamma at 0.05 mg/m2, daily for 6 weeks. One patient from part I and eight patients from part II of the study received three times per week maintenance thereby for up to 14 months. Prior to and at selected times during and after treatment, the clinical and immunologic status of the patients was assessed. RESULTS: In part I, spontaneous de novo IgE synthesis by PBMC was inhibited in 10 patients receiving rIFN-gamma at 0.01 mg/m2 (p = 0.038) and in nine at 0.1 mg/m2 (p = 0.066). There was no reduction of serum IgE levels at any of the three dose levels. Total clinical severity showed improvement at each dose level (p less than 0.04) with worsening 3 days after discontinuation of treatment. In part II, there was no significant inhibition of spontaneous IgE synthesis by PBMC nor was there any reduction of serum IgE. Nevertheless, there was a progressive and significant reduction (p less than 0.01) in total clinical severity over the 6 weeks of daily rIFN-gamma with a sustained improvement during maintenance therapy. CONCLUSION: The results of this pilot study suggest that rIFN-gamma may be efficacious in the treatment of AD and that further clinical trials are warranted.

Adolescent↗
PubMed1989

Generation of hybrid human immunodeficiency virus by homologous recombination.

Human immunodeficiency virus (HIV) type 1, isolated from diverse sources, exhibits genomic diversity. The mechanisms by which the genomic diversity takes place in individuals exposed to multiple virus isolates is yet to be elucidated. Genetic variation, in general, might result from mutagenic events such as point mutations, rearrangements (insertions and deletions), and recombination. In an attempt to evaluate the process of genetic diversity, we designed experiments to analyze recombination between HIV DNAs by using DNA transfection in cell cultures. Here we report the successful recombination between truncated HIV proviral DNAs with an overlap homology of 53 base pairs that leads to the formation of viable hybrid virus. Recombination was also seen between exogenous DNA introduced into cells and homologous HIV sequences resident in the cells. These results indicate that recombination among various HIV isolates may play a significant role in the generation of genetic diversity of HIV. Further, the method used here enables the construction of hybrid HIV genomes to identify the viral determinants responsible for tropism, replication, and cytopathic effects.

Base Sequence↗
PubMed1989

Molecular characterization of HIV-1 isolated from a serum collected in 1976: nucleotide sequence comparison to recent isolates and generation of hybrid HIV.

Human immunodeficiency virus type 1 (Z321 designate, HIV-1Z321), the oldest known HIV, was isolated from a serum sample collected in Zaire in 1976 and was molecularly cloned. Restriction enzyme analysis of unintegrated viral DNA revealed the presence of conserved restriction enzyme cleavage sites in the long terminal repeat sequences. Nucleotide sequence analysis of the 3' end of the viral DNA revealed a pattern similar to other HIV-1 isolates described. However, some of the common restriction sites present in other isolates were absent in HIV-1Z321. The extent of differences between HIV-1Z321 and recent isolates from North America and Zaire was 17.86-18.36% on the nucleotide sequence level and 26.5-33.2% difference in the predicted amino acid sequence in the envelope gene. Differences were also noted in 3'-orf (nef: according to HIV gene nomenclature; see Ref. 42) gene and U3 region of the long terminal repeat sequences of HIV-1Z321 and other isolates. Nucleotide sequence of a HIV-1 isolate, 12 years apart from the present isolates, will provide an important time calibration point for the evolutionary divergence of HIV isolates. Hybrid HIV was also generated by transfecting HIV-1Z321 and HIV-1HTLV-III viral DNAs into cells.

Amino Acid Sequence↗
PubMed1989

Replication of HIV in human fetal retinal cultures and established pigment epithelial cell lines.

The ability of the human immunodeficiency virus type 1 (HIV-1) to replicate in cells derived from ocular tissue was studied. Primary retinal cultures (containing both glial and neuronal cells) were found to support the replication of HIV upon transfection with molecularly cloned proviral DNA. In addition, established retinal pigment epithelial (RPE) cell lines also produced HIV particles upon transfection. HIV released by these cell lines was able to infect and induce characteristic cytopathic effects in T4+ cells. An indicator plasmid containing the HIV long terminal repeat sequences (LTR) linked to the chloramphenicol acetyltransferase gene showed barely detectable activity in RPE cells and was transactivated by the addition of the HIV "tat" gene. Based on these observations, direct infection of ocular tissue derived cells such as RPE, fetal retinal cells, retinoblastoma cells (Y 79, WER1), choroidal endothelial cells (Chor 55) (mix culture) and corneal fibroblasts (K61) by HIV was attempted. HIV replication in these cells was not detected by reverse transcriptase, antigen and transactivation function assays.

Adult↗
PubMed1988

Homologous recombination between human immunodeficiency viral DNAs in cultured human cells: analysis of the factors influencing recombination.

Recombination between HIV DNAs was analyzed using DNA transfection in cell cultures and the optimal conditions for efficient recombination were determined. Recombinant plasmid DNA substrates were constructed from HIV proviral DNAs and the success of recombination was measured by the production of viable hybrid virus. The process of recombination between HIV DNAs was shown to be i) dependent on homology between the truncated HIV DNAs and ii) maximum with concentrations of the truncated DNAs 3ug and above. HIV isolates with heterogeneity in their primary sequence, thus offer an ideal system for the analysis of the requirement of homologous recombination. In addition, recombination methodology would be useful for generating hybrid HIVs for the analysis of specific viral gene functions.

Calcium Phosphates↗
PubMed1988

Studies on human immunodeficiency virus-induced cytopathic effects: use of human rhabdomyosarcoma (RD) cells.

A full-length molecular clone of human immunodeficiency virus (HIV) proviral DNA was transferred to human rhabdomyosarcoma (RD) cells by gene transfer method. RD cells released infectious virus within 12 hours after transfection and the viral particles present in the culture medium could be quantitated by monitoring reverse transcriptase activity. Chronic low level viral producer cell lines of RD were also established. Southern hybridization analysis revealed the presence of HIV sequences in transfected RD cells. Electron microscopic studies of the transfected cell revealed intracellular budding of HIV and also showed structural abnormalities such as giant cell phenotype and vacuolation. These features qualify RD cells as a useful system for studying the regulation and cytopathic effects of HIV.

Cytopathogenic Effect, Viral↗
PubMed1988

Human immunodeficiency virus replication in human brain cells. Brief report.

Human immunodeficiency virus (HIV), the etiologic agent of AIDS, was found to infect and replicate in human brain cells. The extent of HIV replication was minimal in human brain-derived cells in comparison to T4 lymphoid cells. These results suggest that direct infection of glial/neuronal cells by HIV may contribute to the CNS dysfunction frequently observed in HIV infected individuals.

Brain↗
PubMed1987

Sodium butyrate activates human immunodeficiency virus long terminal repeat--directed expression.

To study the effect of sodium butyrate on human immunodeficiency virus (HIV) long terminal repeat (LTR)--directed expression, we constructed a chimeric plasmid (pLTR-CAT) in which the LTR sequences derived from a molecular clone of HIV were fused to the chloramphenicol acetyltransferase (CAT) gene. We used transient expression assays in transfected tissue culture cells to monitor the activity of the LTR. The expression of the pLTR-CAT plasmid was activated when the cells were exposed to butyrate after transfection. The magnitude of butyrate-induced increase was linear up to an 8 mM concentration and was different with regard to the target promoters used. Recombinant plasmids linked to marker genes may be useful models for studying the effects on HIV of various agents of chemical and biological origin.

Butyrates↗
PubMed1987

Characteristics of a novel lentivirus derived from South African sheep with pulmonary adenocarcinoma (jaagsiekte).

A novel lentivirus was isolated from South African sheep with experimentally transmitted lung adenocarcinoma. Similar to visna virus and caprine arthritis encephalitis virus, this new strain induced cytopathic effects on ovine plexus choroid cultures. In contrast to a recent Israeli isolate from sheep with adenocarcinoma, the South African lentivirus could not transform fibroblast cultures. The antigenic relatedness between the new isolate and visna virus was assessed by immunoprecipitation of radiolabeled viral proteins, using monospecific antisera against visna virus proteins. The results indicate that the new virus contains four major structural proteins of sizes similar to those of visna virus (i.e., gp135, p30, p16, and p14) and have some common antigenic determinants (about 90% in the major core antigen p30). However, the nucleotidic sequences of the novel lentivirus were found to be only 16.5 to 27.4% homologous to visna virus and 8.3 to 15% homologous to caprine arthritis encephalitis virus, by means of liquid hybridization under stringent conditions. The genetic divergence indicated by this last result was confirmed by the dissimilar restriction endonuclease cleavage map of the new virus in comparison to those of visna virus and three caprine arthritis encephalitis virus strains. The demonstration of a third type of ovine lentivirus supports the concept of an important genetic variation among the lentiviruses infecting one animal species.

Animals↗
PubMed1987

Molecular characterization of human immunodeficiency virus from Zaire: nucleotide sequence analysis identifies conserved and variable domains in the envelope gene.

To examine the genetic relatedness of human immunodeficiency viruses (HIV) from different geographic locations, we molecularly cloned the genome of HIV isolated from a Zairian AIDS patient. Restriction mapping of the recombinant clone, designated HIV-Zr6, revealed both common (as observed in other HIV isolates) and unique restriction sites. The DNA clone of HIV-Zr6, shown to give rise to infectious cytopathic virus after transfection of cultured lymphoid cells, was sequenced in several regions. The long terminal repeat (LTR), open reading frame 1 (ORF1), C-terminal envelope (env) gene domain, and ORF2 showed less than 6% difference in nucleotide sequence when compared to other HIV isolates including human T-lymphotropic virus-type III (HTLV-III) clone B10, lymphadenopathy-associated virus-1 (LAV-1), and AIDS-associated retrovirus-2 (ARV-2). About 15% difference in nucleotide sequences was noted in the N-terminal env gene domain. Alignments of env gene sequences revealed conserved, moderately variable, and hypervariable stretches in the predicted amino acid sequences. This model provides a basis for assessing the significance of sequence variation on properties controlled by the viral Env glycoproteins such as cell tropism and immunogenicity.

Acquired Immunodeficiency Syndrome↗
PubMed1987

Rodent and primate monolayer cells support the expression and assembly of viral particles directed by human immunodeficiency virus (HIV) proviral DNA.

A molecular clone of human immunodeficiency virus (HIV) was transferred to rodent and primate monolayer cells by DNA transfection. Of the cell lines tested, all of them released viral particles as shown by reverse transcriptase assay and dot-blot hybridization. The viruses released from transfected cultures were able to establish infection of human T-cell line HUT 78. These results suggest that factors present in cells of non-lymphoid origin support the expression and assembly of HIV. Thus, transfection with cloned HIV DNA provides an approach to analyse the consequences of viral gene expression in cell types which lack a mechanism for efficient uptake of HIV particles.

Animals↗
PubMed1987

Transfusion-associated AIDS: donor-recipient human immunodeficiency virus exhibits genetic heterogeneity.

The genetic diversity of the human immunodeficiency virus (HIV) isolated from transfusion-associated AIDS patients has been examined. Restriction enzyme mapping studies of integrated proviral DNA of donor and recipient origin demonstrated genomic variation between isolates. Analysis of the molecularly cloned viral genomes of one donor-recipient pair showed that virus from the recipient had restriction enzyme site differences from the donor, noticeably clustered in the env and orf-2 regions, and also had a greater number of restriction sites in common with the donor as well. These results suggest that HIV may undergo genomic variation in vivo. Comparison of donor-recipient viruses may further the understanding of the molecular basis for AIDS pathogenesis.

Acquired Immunodeficiency Syndrome↗
PubMed1986

Studies on the mechanism of membrane fusion: site-specific mutagenesis of the hemagglutinin of influenza virus.

Oligonucleotide-directed mutagenesis of a cDNA encoding the hemagglutinin of influenza virus has been used to introduce single base changes into the sequence that codes for the conserved apolar "fusion peptide" at the amino-terminus of the HA2 subunit. The mutant sequences replaced the wild-type gene in SV40-HA recombinant virus vectors, and the altered HA proteins were expressed in simian cells. Three mutants have been constructed that introduce single, nonconservative amino acid changes in the fusion peptide, and three fusion phenotypes were observed: substitution of glutamic acid for the glycine residue at the amino-terminus of HA2 abolished all fusion activity; substitution of glutamic acid for the glycine residue at position 4 in HA2 raised the threshold pH and decreased the efficiency of fusion; and, finally, extension of the hydrophobic stretch by replacement of the glutamic acid at position 11 with glycine yielded a mutant protein that induced fusion of erythrocytes with cells with the same efficiency and pH profile as the wild-type protein. However, the ability of this mutant to induce polykaryon formation was greatly impaired. Nevertheless, all the mutant proteins underwent a pH-dependent conformational change and bound to liposomes. These results are discussed in terms of the mechanism of HA-induced membrane fusion.

Animals↗
PubMed1986

Motor evoked potential in cats with acute spinal cord injury.

We have previously reported that a motor evoked potential (MEP) can be produced by transcranial stimulation of the motor cortex in cats and humans. This signal travels in both dorsolateral and ventral spinal cord. We report here the evaluation of this evoked potential in comparison to the somatosensory evoked potential (SEP) in an acute spinal cord weight drop model. In all animals, the peripheral nerve signal was the component of the MEP most sensitive to injury. Often, it was significantly reduced in amplitude by incidental manipulation of the spinal cord during a careful laminectomy and then returned. It was lost first in animals with weight drop spinal cord injury and was abolished with as little as 50 to 75 g/cm of force. The spinal cord signal of the MEP was consistently more sensitive to injury than the SEP and was abolished at about 100 to 150 g/cm of impact. The cortical SEP was abolished at about 200 to 250 g/cm, and the spinal cord SEP was abolished at similar levels. The SEP returned earlier after injury than the MEP. Anesthetic agents had an effect on the MEP in the spinal cord and substantially changed the peripheral nerve signal, in both wave form shape and optimal stimulation frequency. Marginal cord injury and abnormal metabolic conditions caused the peripheral nerve signal to decrement in amplitude with increasing trial numbers during a run and become unstable. These latter effects need further characterization and are critical guides to investigative and clinical use of this test. This study indicates that the MEP is more sensitive than the SEP in detecting spinal cord injury.

Animals↗