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Biomedical subjects

L J Ross

Publications and source records attributed to L J Ross.

At least 37 records · Page 2Linked to original sources

African-American women and abortion: a neglected history.

The history of African-American women's efforts to control their fertility is largely unknown. From slavery to the present, the growth rate of the African-American population has been cut in half. Demographers and historians frequently attribute this change to external factors such as poverty, disease, and coerced birth control, rather than the deliberate agency of African-American women. This essay assembles a brief historical record of the ways African-American women have sought to control their fertility through the use of abortion and birth control. It also examines the activism of African-American women in the establishment of family planning clinics and in defense of abortion rights.

Abortion, Illegal↗

Mechanism of inhibition of human immunodeficiency virus type 1 reverse transcriptase and human DNA polymerases alpha, beta, and gamma by the 5'-triphosphates of carbovir, 3'-azido-3'-deoxythymidine, 2',3'-dideoxyguanosine and 3'-deoxythymidine. A novel RNA template for the evaluation of antiretroviral drugs.

Carbovir (the carbocyclic analog of 2'-3'-didehydro-2',3'-dideoxyguanosine) is a potent inhibitor of human immunodeficiency virus type 1 (HIV-1) replication. Assays were developed to assess the mechanism of inhibition by the 5'-triphosphate of carbovir of HIV-1 reverse transcriptase using either RNA or DNA templates that contain all four natural nucleotides. Carbovir-TP was a potent inhibitor of HIV-1 reverse transcriptase using either template with Ki values similar to that observed by AZT-TP, ddGTP, and ddTTP. The kinetic constants for incorporation of these nucleotide analogs into DNA by HIV-1 reverse transcriptase using either template were similar to the values seen for their respective natural nucleotides. In addition, the incorporation of either carbovir-TP or AZT-TP in the presence of dGTP or dTTP, respectively, indicated that the mechanism of inhibition by these two nucleotide analogs was due to their incorporation into the DNA resulting in chain termination. Carbovir-TP was not a potent inhibitor of DNA polymerase alpha, beta, or gamma, or DNA primase. Given the potent activity of carbovir-TP against HIV-1 reverse transcriptase and its lack of activity against human DNA polymerases, we believe that further evaluation of this compound as a potential drug for the treatment of HIV-1 infection is warranted.

Antiviral Agents↗

A TIBO derivative, R82913, is a potent inhibitor of HIV-1 reverse transcriptase with heteropolymer templates.

R82913, (+)-S-4,5,6,7-tetrahydro-9-chloro-5-methyl-6-(3-methyl-2-butenyl)- imidazo[4,5,1-jk][1,4]-benzodiazepin-2(1H)-thione (a TIBO derivative), inhibited the replication of thirteen different strains of HIV-1 in CEM cells with a median IC50 of 0.15 microM. The concentration of compound that killed 50% of the cells was much higher (46 microM), indicating that R82913 has a high selectivity index. R82913 was 20-fold more potent than AZT-TP in the inhibition of HIV-1 reverse transcriptase in an assay using a naturally occurring template (ribosomal RNA) that more accurately resembles native viral RNA than a synthetic homopolymer. With this template, R82913 inhibited HIV-1 reverse transcriptase with an ID50 (0.01 microM) that is equal to, or lower than, the IC50 for this compound in all of our cell culture assays (0.01-0.65 microM). R82913 has no effect on the replication of HIV-2 in CEM cells and does not inhibit the reverse transcriptase from this virus.

Base Sequence↗

Properties and evolutionary relationships of the Marek's disease virus homologues of protein kinase, glycoprotein D and glycoprotein I of herpes simplex virus.

The deduced amino acid sequences of the open reading frames (ORFs) mapping in the short unique segment (US) of Marek's disease virus (MDV) reported in the accompanying paper have been analysed using computer programs to determine their relationships to herpesvirus proteins. Analysis of the catalytic domains of protein kinases showed that the MDV kinase (MDV PK) was closely related to the alphaherpesvirus protein kinase mapping in US. The results also showed that the MDV PK was more closely related to the cellular kinases that control cell division than to the proto-oncogenes c-src and c-mos and it was predicted that the MDV PK would phosphorylate serine/threonine. The MDV homologue of herpes simplex virus (HSV) glycoprotein D (gD) contained several residues that were conserved in mammalian herpesviruses. In particular, six cysteines were perfectly aligned in all the gDs and there were numerous conservative substitutions. Although only approximately 65% of the MDV homologue of glycoprotein I (gI) of HSV has been sequenced, it was clear that a significant number of amino acid residues including four cysteines were conserved in the gI homologues of MDV and mammalian herpesviruses. Further analysis suggested that MDV gD was more closely related to the gDs of pseudorabies virus (PRV) and equine herpesvirus 1 than to the gD of HSV-1 and HSV-2. It was noted that HSV-2 glycoprotein G (gG), PRV gX and MDV gD were related and that MDV ORF4 was related to MDV gD and probably to HSV-1 gG. The results have shown a clear relationship between the genes of MDV and their counterparts in mammalian alphaherpesviruses and are consistent with the idea that MDV glycoprotein genes in US might have arisen by a process of gene duplication and independent evolution.

Amino Acid Sequence↗

DNA sequence and organization of genes in a 5.5 kbp EcoRI fragment mapping in the short unique segment of Marek's disease virus (strain RB1B).

The DNA sequence of a 5.5 kbp EcoRI fragment located in the short unique region (US) of the 'highly oncogenic' strain RB1B of Marek's disease virus (MDV) was determined. The sequence contained six open reading frames (ORFs), four of which were homologous to proteins mapping in the US region of herpes simplex virus type 1 (HSV-1). These include the homologues of HSV-1 protein kinase, glycoprotein D (gD), glycoprotein I (gI) and US2 which is of unknown function. The MDV ORFs had a marked bias for A or T in the third codon position and analysis of the dinucleotide frequencies showed a marginal deficit in ApG/CpT but no overall deviation of CpG from random expectations. Comparison of genes in the US region of MDV to herpesvirus proteins confirmed and extended our previous observation that MDV is more closely related to alphaherpesviruses than to gamma-herpesviruses. We also showed that MDV possessed a homologue of HSV-1 gD which is lacking in varicella-zoster virus (VZV) but that MDV probably lacked homologues of US4 and US5 of HSV-1. These results show that in contrast to the genes in the long unique region which were grossly collinear in HSV, VZV and MDV, those mapping in US show greater diversity.

Amino Acid Sequence↗

Partial transcription map of Marek's disease herpesvirus in lytically infected cells and lymphoblastoid cell lines.

Marek's disease herpesvirus (MDV) can cause either a productive-restrictive or lytic infection, a latent infection or can transform thymus-derived lymphocytes. RNA was extracted from infected chicken embryo fibroblasts (CEF) or from lymphoblastoid tumour cell lines. Some of the infected CEF were treated with 200 micrograms/ml cycloheximide to identify immediate early (IE) transcripts, and others with 1 microM 1-(2-fluoro-2-deoxy-B-D-arabinofuranosyl)-5-methyluracil (FMAU), an inhibitor of herpesvirus DNA synthesis to identify early transcripts. An extensive Northern blot analysis was carried out using DNA probes spanning almost the complete MDV genome. In the lytically infected CEF at least 66 discrete transcripts were detected, ranging in size from 9.1 kb to 0.6 kb. Eleven IE transcripts were identified, of which 8 were mapped in the genome segment consisting of the IRL, IRS, US and TRS. Six transcripts were identified as early genes. In the MD lymphoblastoid cell lines MDCC-HPI, a non-producer cell line, and MDCC-CU41, a non-expression cell line, 4 and 7 transcripts were detected, respectively. These RNAs were transcribed from IE genes located mainly in the repeat sequences flanking UL and US and in US. Treatment of the lymphoblastoid cell lines with 20 micrograms/ml 5-iodo-2-deoxyuridine resulted in the additional transcription of 1 RNA species in HPI and 9 in CU41. Most of the transcripts present in lytically infected cells were also detected in MDCC-CU36, a cell line with a high percentage of antigen-positive cells (expression cell line).

Cell Line↗

Nucleotide sequence and characterization of the Marek's disease virus homologue of glycoprotein B of herpes simplex virus.

The Marek's disease virus (MDV) homologue of the herpes simplex virus (HSV) gene encoding glycoprotein B (gB) has been identified within BamHI fragments I3 and K3 of the 'highly oncogenic' strain RB1B of MDV. The entire nucleotide sequence of the gene has been determined and its predicted amino acid sequence shown to share gross overall structural features with the gB genes of HSV, varicella-zoster virus (VZV) and other mammalian herpesviruses. In particular, all 10 cysteine residues were conserved in MDV gB and there was extensive homology throughout the gene with VZV, HSV and pseudorabies virus except for the N and C termini. The overall percentage amino acid identity between MDV gB and gB of the alphaherpesviruses had a mean of 50% which was almost twice that between cytomegalovirus and Epstein-Barr virus. Northern blot analysis showed that the main RNA transcribed from this gene is approx. 2.7 kb in size. Antibodies raised against synthetic peptides (residues 250 to 271 and 304 to 330) allowed the identification of a family of serologically related glycoproteins of Mr 110K, 64K and 48K in extracts of MDV-infected cells using immunoblots. Furthermore, the antisera were able to differentiate between the antigens of MDV and herpesvirus of turkeys in immunoblots. Immunofluorescence tests indicated that MDV gB is associated with granules in the cytoplasm and is present at the surface of MDV-infected cells.

Amino Acid Sequence↗

Further characterization of Marek's disease virus-infected lymphocytes. I. In vivo infection.

Previous reports from this laboratory identified bursa-derived lymphocytes (B cells) and non-B cells as the predominant cell types respectively involved in the early cytolytic and subsequent latent infection of chickens with Marek's disease virus (MDV). It was not known whether these differences were qualitative or quantitative or if the method for detection of latent infection (viral antigen production after 48 h of in vitro cultivation) was sensitive enough. To further define the cells involved in the various phases of MDV infection, we used monoclonal antibodies which specifically react with B cells, or T cells, or la-antigen-bearing cells. Dual fluorescence tests to detect surface markers and viral internal antigen (VIA) were conducted with infected spleen cells freshly collected from MDV-infected chickens or after in vitro cultivation of those cells. The same antibodies were also used for a rosetting procedure to yield fractions enriched or depleted of T cells, B cells or la-bearing cells. These were examined directly for viral DNA by in situ hybridization or dot blot DNA hybridization and for VIA cultivation. We learned that infected T cells also comprise part of the early cytolytic phase of MDV infection but constitute a minority population (approximately 2-3%) compared to B cells (83-92%) at 3 or 4 days post infection. Latently infected cells were definitively identified as mostly la-bearing T cells, although a few (2-4%) were B cells. Prior to in vitro cultivation, latently infected cells apparently had insufficient viral DNA for detection by in situ hybridization, but the more sensitive dot blot procedure revealed viral DNA in fractions later found positive by VIA expression after in vitro cultivation. Viral DNA replication in latently infected cells apparently had occurred after 48 h cultivation because in situ hybridization detected infected cells at that time. Treatment of cell cultures with iodo-deoxyuridine, 12-O-tetradecanoyl phorbol-13-acetate or n-butyrate failed to increase the number of spleen cells which expressed VIA.

Animals↗

Further characterization of Marek's disease virus-infected lymphocytes. II. In vitro infection.

Lymphocyte cultures from chicken spleens had been shown to be susceptible to in vitro infection by Marek's disease virus (MDV)4 in an earlier report from this laboratory. In that study, virus infection was evidenced by virus isolation and detection of viral internal antigen (VIA) 2 days post inoculation (DPI), and serial passage was accomplished by adding fresh spleen cells at 2-day intervals. The susceptible cells were identified as bursa-derived lymphocytes (B cells). Using a dual fluorescence technique to identify surface markers for B cells, thymus-derived lymphocytes (T cells) or Ia antigen on VIA-positive cells, we have now shown that a small proportion (generally less than 10%) of VIA-positive cells observed 2 DPI are T cells, and that a low level of infection can be maintained by serial passage of MDV in cultures totally free of B cells. Most infected T cells in this study had Ia antigen. As the incubation period for infected cultures was extended from 2 to 4 or 5 days, the average number of viable cells decreased but the percentage of viable cells infected with MDV (VIA-positive) increased. Also, both the proportion and the actual number of infected T cells increased, significantly more so in cultures from genetically susceptible P-2 donors than from resistant N-2 donors. Spleen-cell cultures from resistant Line 6 chickens were markedly less susceptible than those from susceptible Line 7 chickens to in vitro MDV infection, as assessed by numbers of VIA-positive cells at 5 DPI.

Animals↗

Restriction endonuclease analysis of Marek's disease virus DNA and homology between strains.

The restriction endonuclease patterns of viral DNA obtained from serotypes 1 and 2 strains of Marek's disease virus have been compared and homology between the strains examined by hybridization. The results have shown that HPRS 16 (serotype 1) DNA has a structure similar to its attenuated variant HPRS 16/att except for a few fragments that are present only in the virulent strain. Evidence was obtained which suggested that these restriction fragments contained repeat sequences and that insertion of heterogeneous DNA occurred at these sites during attenuation of HPRS 16. The restriction enzyme patterns of HPRS 24 (serotype 2) differed substantially from those of HPRS 16 and HPRS 16/att and reassociation experiments showed that HPRS 24 shares less than 10% homology with either HPRS 16 or herpesvirus of turkeys (serotype 3).

Base Sequence↗

Susceptibility of Marek's disease lymphoblastoid cell lines to infection with influenza and pseudorabies viruses and the protective effect of immunization with influenza virus-infected lymphoblastoid cells.

The susceptibility of 3 lymphoblastoid cell lines (MDCC-MSB1, MDCC-HP1 and MDCC-RP1) derived from Marek's disease tumours to infection with influenza (AoPR8) and pseudorabies viruses was studied. MSB1 and HP1 were more susceptible to infection with influenza virus than was RP1. It was shown in the case of HP1, which was studied in greater detail, that although the majority of the cells synthesized influenza virus haemagglutinin when infected at high multiplicity, only a small proportion produced mature infectious particles. The results obtained from both high and low multiplicities of infection suggested that infection with AoPRs was abortive in the majority of the cells of all 3 cell lines. Infected cultures remained viable when subcultured over several days. All 3 cell lines were resistant to pseudorabies virus. The block in pseudorabies virus replication probably occurred after adsorption. Immunization of chickens with HP1 lymphoblastoid cells superinfected with influenza virus AoPR8 and inactivated with glutaraldehyde gave better protection against Marek's disease than immunization with uninfected HP1 cells.

Animals↗

Characterization of an antigen associated with the Marek's disease lymphoblastoid cell line MSB-1.

A Marek's disease lymphoblastoid cell line (MSB-1) has been analysed by immunoprecipitation for expression of tumour-associated antigen, Marek's disease virus (MDV)-specific antigens and antigens specific to avian leukosis-sarcoma viruses. Rabbit antisera raised against two independently derived cell lines after extensive absorption with normal chick cells reacted with a polypeptide of mol. wt. 40 000 (40K) in extracts of MSB-1 cells. The 40K polypeptide was not present in myeloblasts or in chick embryo fibroblasts (CEF) infected with MDV and did not react with antiserum raised against normal chicken thymus antigens. The possibility that the 40K polypeptide is a tumour-associated antigen is discussed. Seven MDV-specific antigens were noted in infected CEF (mol. wt. 110K, 100K, 80K, 70K, 50K, 35K and 32K) but none of these was detected in MSB-1 cells. The avian leukosis-sarcoma group-specific antigen P27gag and its precursor Pr76gag were not found in MSB-1 cells, confirming that expression of mature gag protein is not required for transformation by MDV. However, two polypeptides of unknown origin and function (mol. wt. 180K and 110K) were precipitated from MSB-1 cells with a rabbit anti-Rous sarcoma (Schmidt-Rupin, subgroup D) antiserum.

Alpharetrovirus↗

Trisomy after pregestational contact with communicable disease.

Six hundred nineteen mothers who had had household contact with rubella, infectious hepatitis, measles, mumps or varicella at a known time before or after their last menstrual periods were identified in a study in West Jerusalem, Israel. The mother's subsequently-born babies included four with Down's syndrome and one with trisomy 18. The mothers of these five children had been exposed to a communicable disease during the five months preceding pregnancy. The possibility that maternal pregestational contact with communicable disease might have contributed to autosomal trisomy in the official offspring is considered.

Adult↗

Nature of genetic resistance to Marek's disease in chickens.

Infection of line 6 resistant and line 7 susceptible chickens with Marek's disease virus (MDV) resulted in a depressed phytohemagglutinin (PHA) response and the presence of Marek's disease (MD) tumor-associated surface antigen (MATSA) in the spleens. At 6-10 weeks after infection, recovery in PHA response, diminution in the number of MATSA cells, and the presence of significant anti-MATSA immunity were observed in line 6 but not in line 7 chickens. Both lines had antibody-dependent cell-mediated antiviral immunity, but T-cell-mediated antiviral immunity was detected only in line 6 and the surviving line 7 chickens. Lymphoproliferative lesions were found only in line 7 chickens, an virus titers were significantly higher in line 7 than in line 6 chickens and embryos. Lymphoid organ weights and the number of lymphocytes of line 6 were significantly lower than those of line 7. These data suggests that resistance to MD in line 6 chickens was due to a) a deficiency in the aggregate number of target lymphocytes, b) a restriction in the ability to lymphocytes to nonproductively replicate MDV, and c) the involvement of cellular antiviral and antitumor immune responses.

Animals↗

The effect of bursectomy on the adoptive transfer of resistance to Marek's disease.

Immunization with inactivated viral antigens protected chickens against Marek's disease. Non-immunized chickens could be protected by injections of spleen cells but not of serum from immunized, histocompatible donors. Chickens rendered agamma-globulinaemic by bursectomy and irradiation could also be immunized against Marek's disease by inoculation with viral antigens, but spleen cells from these immunized, bursectomized and irradiated donors did not confer protection on the recipients into which they were injected. It was concluded that, although in the bursectomized, immunized donors cell-mediated immunity alone was able to provide a fair degree of protection against Marek's disease, the protection afforded against the disease by spleen-cell transfer was at least partly attributable to the transfer of antibody-producing cells, and that humoral immunity, while not being an absolute requirement for resistance, is normally an important component of the resistance mechanism.

Animals↗

The effects of ultraviolet and ionizing radiation on herpesviruses, SV40 and adenoviruses in relation to the small-plaque effect.

The small-plaque effect occurs with a wide range of herpesviruses following irradiation with ultraviolet light. The 37 per cent survival (D37) values, or dose required for one lethal hit (e-1), for herpes simplex, pseudorabies and pigeon herpesviruses in different cells indicate a broad spectrum of host-cell repair capacity. Other DNA-containing viruses such as SV40 and adenoviruses, which also replicate in the cell nucleus, show the small-plaque effect. Ionizing irradiation of herpes simplex virus type 1 (HSV-1) showed but little reduction in plaque-size.

Adenoviruses, Human↗

Fingerprints in congenital rubella following maternal gamma globulin.

The fingertip skin ridge patterns of ten children with the congenital rubella syndrome whose mothers had received gamma globulin following exposure to german measles during pregnancy were compared with those of 29 patients whose mothers had not received gamma globulin and with those of 162 control children. Statistical evaluation of the pattern profiles by the Mann-Whitney U-test showed that the dermatoglyphics of the two groups of patients differed significantly from each other. Each group also differed from the control children: there was a significant (p less than 0.001) increase in whorls in patients whose mothers had not had gamma globulin, but an increase (p = 0.001) in ulnar loops in those whose mothers had received gamma globulin.

Dermatoglyphics↗