PubMed Health⌕ Search

Biomedical subjects

Y Becker

Publications and source records attributed to Y Becker.

At least 91 records · Page 5Linked to original sources
PubMed1989

Ataxia-telangiectasia fibroblasts have less fibronectin mRNA than control cells but have the same levels of integrin and beta-actin mRNA.

The expression of fibronectin, integrin and beta-actin genes in skin fibroblasts from patients with the genetic disorder ataxia-telangiectasia (A-T) was studied. These three genes were selected because their protein products contribute to the shape and function of the fibroblast. Expression of mRNA by these genes was compared with that in fibroblasts from normal individuals and from patients with the genetic disorder, hereditary hemorrhagic telangiectasia (HHT). A-T fibroblasts were found to produce less fibronectin mRNA than normal and HHT fibroblasts. A-T fibroblasts senesce around passage level 15 while normal and HHT fibroblasts can be propagated for many more passages in vitro. However, the expression of the integrin gene in A-T fibroblasts was similar to that in normal fibroblasts, while the beta-actin gene was expressed at a higher level. The increased beta-actin mRNA levels were similar in fibroblasts of patients with the two genetic disorders, A-T and HHT, but higher than in normal fibroblasts. HHT fibroblasts differed markedly from A-T fibroblasts in having a high level of fibronectin and integrin mRNA expression. The results indicate that regulation of the fibronectin gene in A-T fibroblasts differs from that of the integrin and beta-actin genes, and that the decline in fibronectin mRNA may be linked to the shortened in vitro life-span of these cells.

Actins↗
PubMed1989

Langerhans cell density and activity in mouse skin and lymph nodes affect herpes simplex type 1 (HSV-1) pathogenicity.

Langerhans cells are epidermal antigen-presenting cells that function by taking up antigens in the skin, migrating to the lymph nodes, where they are designated interdigitating cells, and triggering the immune response. The role of interdigitating cells (IDC) was investigated in a murine model of herpes simplex virus-1 infection in the skin. The number of IDC in the lymph nodes began to increase on the first day following infection and reached a peak three days p.i. Low titers of infectious virus were recovered from the fraction of lymph node cells that consisted of 60-80% IDC at one day p.i. Lymph node cells that were obtained from mice immunized with HSV-1 proliferated in vitro in response to viral antigens but did not respond to mock antigens. When mice were immunized with HSV-1 inoculated into skin that had been depleted of Langerhans cells, this in vitro proliferative response was abolished. Thus, the present results suggest that Langerhans cells function in the immune defense of the skin against HSV-1 infection by transporting the virus to the peripheral lymph nodes where an immune response is initiated. Injection of the immunomodulator OK-432 into the footpad skin caused a local increase in the number of Langerhans cells in the epidermis and led to an increased migration of dendritic cells to the lymph nodes. Under these conditions, a decrease in HSV-1 pathogenicity was noted. These observations indicate that the pathogenicity of herpes simplex virus type 1 in the skin is affected by Langerhans cell density and activity in the epidermis and the lymph nodes.

Adenosine Triphosphatases↗
PubMed1989

Langerhans cells in vaccinia virus infection in mouse skin.

Langerhans cells function as potent antigen-presenting cells in the epidermis. They were shown to play an essential role in the mechanisms of defense of the skin against viral infections. In the present study, the response of Langerhans cells to infection of the skin with vaccinia virus was investigated. Decrease in Langerhans cell density in the skin was accompanied by an increase in the pathogenicity of the WR and Noguchi but not of the Lister strain of vaccinia virus. Langerhans cell density was shown to increase rapidly at the site of inoculation with the two pathogenic strains of vaccinia virus.

Animals↗
PubMed1989

Characterization of RNA transcripts from herpes simplex virus-1 DNA fragment BamHI-B.

Herpes simplex virus type 1 (HSV-1) DNA fragment BamHI-B (0.738-0.809 map units) was recently reported to be associated with the phenotype of intraperitoneal pathogenicity and to encode a latency-associated RNA transcript. Part of this fragment resides within the internal repeat sequence of the long (L) region of the viral genome. In this study, RNA transcripts from BamHI-B were characterized. In addition to immediate-early mRNAs IE-1 and IE-2, eight novel RNA species were found. Three transcripts were mapped in the repeat regions of this fragment and five transcripts in the unique L region of BamHI-B. In addition, transcription activity from these regions was compared in several HSV-1 strains. These included the intraperitoneal virulent F and KOS strains, the avirulent strain HFEM, as well as the HFEM/F intratypic virulent recombinant R-MIC1. Several differences were noted and their possible relevance to virulence is discussed.

Animals↗
PubMed1989

Cytofluorometric analysis of thymic interdigitating cells from C57BL/6 mice prior and after leukemogenic X-irradiation.

The thymus is populated by various Ia+ cell populations, including epithelial cells, macrophages and dendritic cells. Thymic cell suspensions were stained with an anti-Ia antibody and shown by cytofluorometry to contain a small number of strongly Ia+ cells characterized by a large diameter. The cell population was separated with the aid of the fluorescence-activated cell sorter (FACS) and characterized. They were shown to express high levels of membranal Ia antigens; they demonstrated ATPase activity and displayed the ultrastructural features characteristic of the previously described thymic interdigitating cells. C57BL/6 mice were submitted to various regimens of X-irradiation. Whereas exposure to a single dose of X-irradiation was followed by an increase in the percentage of strongly Ia+ cells, exposure to a leukemogenic regimen of fractionated X-irradiation led to a decrease in the percentage and absolute numbers of these cells in the thymus. Of the C57BL/6 mice that were irradiated with fractionated X-irradiation, 77% developed leukemia. Intravenous injection of syngeneic bone marrow one day following the last irradiation or protection of the femur during irradiation prevented both the appearance of leukemia and the disappearance of interdigitating cells. Therefore an inverse correlation between the presence of thymic dendritic cells and the incidence of leukemia in C57BL/6 mice could be demonstrated. These findings are discussed in relation to the putative role of dendritic cells in the thymus.

Animals↗
PubMed1989

Skin fibroblasts from patients with the genetic disorder hereditary hemorrhagic telangiectasia compared with ataxia-telangiectasia fibroblasts in their response to the radiomimetic drug neocarzinostatin.

Skin fibroblasts from patients with hereditary hemorrhagic telangiectasia (HHT) were compared with fibroblasts from patients with the genetic disorder ataxia-telangiectasia (A-T), and with SV40-transformed A-T fibroblasts, regarding their sensitivity to the radiomimetic drug neocarzinostatin (NCS). Whereas A-T fibroblasts were found to be hypersensitive to NCS at low concentrations (as measured by the cellular survival test), as previously reported, the HHT fibroblasts were more resistant to NCS and behaved as normal fibroblasts. The SV40-transformed A-T cells also resembled normal fibroblasts in their response to NCS in the colony formation test. In the DNA synthesis test, HHT strains of fibroblasts did not fully resemble untransformed and SV40-transformed A-T cells that continued to synthesize DNA following NCS treatment, since NCS inhibited DNA synthesis in HHT fibroblasts by 5 to 15%; whereas the same treatment in normal fibroblasts reduced DNA synthesis by 40%.

Antibiotics, Antineoplastic↗
PubMed1989

Computer modeling of membrane-anchored cellular and viral proteins: organization and function.

1. Although secondary structure predictions are only 60% correct, the computer programs provide some idea as to peptide conformation. 2. The detection of hydrophobic domains in membrane-bound proteins allowed the identification of membrane insertional domains in the polypeptide. 3. A combination of computer analyses of protein conformation and the identification of functional domains in polypeptides must be substantiated by experimental data. 4. Computer analyses of membrane proteins may provide a theoretical approach to the selection and development of antibiotics capable of interfering with the insertion and function of viral proteins in cellular membranes.

Algorithms↗
PubMed1988

Computer-assisted primary and secondary structure analyses of DNA polymerases of herpes simplex, Epstein-Barr and varicella zoster viruses reveal conserved domains with some homology to DNA-binding domain in E. coli DNA pol I.

The primary and secondary structure of herpes simplex virus type 1 (HSV-1), varicella-zoster (VZV) and Epstein-Barr virus (EBV) DNA polymerases was calculated by means of computer programs. The comparison of HSV-1 polymerase (pol) sequence to the known primary and tertiary structure of E. coli DNA pol I revealed five short homologous sequences, one of which coincided with the alpha-helical structure of the DNA-binding domain of E. coli DNA pol. Comparison by primary and secondary structure computer programs of the three DNA polymerases coded by herpesviruses HSV-1, VZV and EBV led to the identification of polypeptide sequences shared by the three DNA pols. In a similar way, the secondary structure of the DNA pol polypeptide in the vicinity of the mutation leading to PAA resistance in HSV-1 DNA pol helped to identify the role of this sequence in the binding of phosphate donated by the nucleoside triphosphate molecule which binds to the DNA pol. Although the computer secondary structure programs are about 60% accurate, it was possible to obtain new information on the properties of certain domains in the DNA polymerases of HSV-1, VZV and EBV.

Amino Acid Sequence↗
PubMed1988

Heterogeneity of BamHi DNA fragments B and E in several HSV-1 strains and recombinants.

The restriction cleavage sites of the BamHI-B and BamHI-E DNA fragments of several Herpes simplex virus type 1 (HVS-1) strains were mapped. These fragments are situated at the ends of the long unique regions and share homologous sequences in the repeat components (TRL and IRL) of the genome. All the strains analyzed were found to have deletions in the Hpal-P fragment, situated in the BamHI-B fragment. Five strains were further analyzed and the deletions were located in the Smal-A fragment (within the Hpal-P fragment). The BamHI-E fragment of four recombinants (obtained by recombination between the HFEM genome and the BamHI-B fragment or part of it from the HSV-1 F strain) were almost identical but differed from another strain [NIH(LP)]. Comparison of the BamHI-B and the BamHI-E fragments of the same strain revealed that the fragments were not identical in all cases.

Cloning, Molecular↗
PubMed1988

Marek's disease virus serotype-1 antigens A and B and their unglycosylated precursors detected by Western blot analysis of infected cells.

The antigenic profile of cell cultures infected with Marek's disease virus (MDV) was determined by the immunoblotting method using convalescent immune serum obtained from chickens that survived infection with MDV strain GA5. The MDV antigen profile in infected cell lysates could be accurately determined since this method has advantages over the immunoprecipitation method used in other studies. We studied six very virulent MDV isolates and the prototype of serotype 1 MDV, the GA5 strain. Immunoblots of NaDodSO4-polyacrylamide gel electrophoresis (PAGE) performed under reducing conditions revealed a main viral antigen (B) of 120-130 kD, which was present in all cell lysates infected with MDV isolates. Analysis of infected cell proteins by NaDodSO4-PAGE performed under nonreducing conditions, revealed a 205 kD major MDV antigen, which, under reducing conditions, was identical to the 130 kD major antigen. The unglycosylated precursors of the major MDV antigens were elucidated. Two polypeptides of 43 and 45 kD were found to be the unglycosylated precursors of MDV antigen A (the glycosylated form of which appears in 4 distinct bands). The unglycosylated precursors of the MDV major antigen B were found to be three polypeptides of 80, 110, and 125 kD.

Animals↗
PubMed1988

RNA transcripts of Marek's disease virus (MDV) serotype-1 in infected and transformed cells.

RNA was isolated from two strains of Marek's disease virus (MDV-Z and MDV-B). The virus was grown in duck embryo fibroblasts (DEF) for 96 hr, 72 hr in the presence of phosphonoacetic acid (PAA) and 24 hr in the presence of cycloheximide added at the time of infection. With the use of DNA probes representing about 80% of the MDV genome, an extensive Northern blot analysis of the RNA was carried out. A similar analysis was done with RNA extracted from the MDV-transformed cell line MSB-1. This study revealed 42, 25 and 29 discrete viral RNA transcripts in MDV-Z and MDV-B-infected DEF and in the MSB-1 cell line, respectively, ranging in size from 0.8 to 13 kb. In MDV-Z-infected DEF, there were twelve late RNA species, two early and eight immediate-early viral transcripts. In MDV-B-infected DEF there were eleven late RNA species, two early and seven immediate-early viral transcripts. The RNA species were homologous for all the probes used except the BamHI-G DNA fragment where no RNA transcripts were detected in the MSB-1 cell line. The RNA transcripts were used to produce a preliminary viral RNA map. Comparison of the location and sizes of the viral RNA transcripts in MDV-infected and MDV-transformed cells revealed several differences.

Animals↗
PubMed1988

Role of epidermal Langerhans cells in viral infections.

Langerhans cells function as highly potent antigen-presenting cells in the epidermis. In the last few years, their role in viral infections has been studied in various experimental systems. They have been shown to be involved in the pathogenesis of a number of infections of viral origin. These include vaccinia virus, human papilloma virus, herpes simplex virus, foot and mouth disease virus and human retrovirus infections. Studies on the effect of various factors, that are known to modulate the activity and density of Langerhans cell in the epidermis, may lead in the future to the development of new strategies aimed at inhibiting virus infections or even eradicating latent infection.

Animals↗
PubMed1988

Role of Langerhans cells and Thy. 1+ effector cells in herpes simplex virus-1 infection in the skin of newborn mice.

Langerhans cells function in the epidermis as very potent accessory cells. Their role as antigen-presenting cells in the immune response following herpes simplex virus type 1 (HSV-1) skin infection of newborn mice was studied. Newborn C 57 BL/6 mice were found to be susceptible while adult mice are resistant to HSV-1 infection in the skin. Because the immune response to HSV-1 infection in the skin is mainly cell-mediated, and therefore dependent on the presence of functional accessory cells, the state of Langerhans cells in the skin of newborns was studied. Staining of whole epidermal mounts revealed similar numbers of Ia+ and ATPase+ Langerhans cells in the epidermis of newborn and adult mice. In a skin lymphocyte reaction assay, Langerhans cells derived from newborn mice were shown to stimulate proliferation of T cells derived from adult allogeneic mice to the same degree as adult-derived Langerhans cells. HSV-1 was injected into the skin of C 57 BL/6 newborn mice together with various preparations of adult-derived spleen cells devoid of antigen-presenting cells. The injected adult-derived lymphocytes were found to confer protection against HSV-1 infection in newborn mice, despite the lack of detectable antigen-presenting cell (APC) function in this cell preparation. The cell subset involved in the transfer of resistance was found to be Ia- and Thy. 1+.

Animals↗
PubMed1988

HSV-1 DNA sequence determining intraperitoneal pathogenicity in mice is required for transcription of viral immediate-early genes in macrophages.

The relationship between intraperitoneal (ip) pathogenicity in vivo of herpes simplex virus type 1 (HSV-1) and infection of macrophages (m phi) in vitro was studied. The apathogenic HSV-1 strain HFEM disappeared from the peritoneum of infected mice following ip inoculation, while the pathogenic F strain persisted in the peritoneum and penetrated the mouse nervous system, and eventually the mice died, showing severe neurological signs. When peritoneal m phi were infected in vitro, a direct correlation with pathogenicity in vivo was found with several HSV-1 strains and recombinants. HSV-1 strains (F, KOS, R-M1C1) which were pathogenic for mice by the ip route, induced cytopathic effect (CPE) in m phi in vitro. Strain F transcribed viral immediate-early genes and synthesized viral DNA in m phi that were treated with L-cell conditioned medium (as a source of colony-stimulating factor) prior to infection. Apathogenic HSV-1 strains (HFEM, R-15, R-19) did not cause CPE in m phi. The HFEM strain was already blocked in the transcription of viral alpha genes in the infected m phi, but replicated well in control BSC-1 cells. An intratypic recombinant (R-M1C1), produced by cotransfection of HFEM DNA with a cloned Mlul-Mlul DNA fragment (coordinates 0.7615-0.796) from HSV-1 strain F, that was shown [Becker et al. (1986). Virology 149, 255-259] to have regained partial ip virulence for mice, now transcribed alpha genes, synthesized viral DNA, and induced CPE in m phi. It appears that the viral DNA fragment responsible for ip virulence is involved in tissue-specific recognition of virus by infected m phi, a function necessary for transcription of viral alpha genes.

Animals↗
PubMed1988

Comparative analysis of the transcripts mapped in the BamHI DNA fragment B of avirulent HSV-1 HFEM, virulent HSV-1 F, and their intratypic recombinant viruses.

HSV-1 HFEM, whose genome harbors a deletion of 4.1 kbp (0.762 to 0.789 map units (mu] is avirulent for mice and tree shrews by the intraperitoneal (i.p.) application route. Insertion of the BamHI DNA fragment B (0.738 to 0.809 mu) and/or the MluI DNA fragment (0.7615 to 0.796 mu) molecularly cloned from virulent HSV-1 F, restored the i.p. pathogenicity to strain HFEM and led to the isolation of virulent intratypic recombinants. In order to determine the RNA transcripts mapped in the BamHI DNA fragment B of the HSV-1 HFEM, HSV-1 F, and their intratypic recombinants R15, R19, R26, and R-Ml-C1, a comparative analysis was performed using Northern blot hybridizations. Two novel RNA transcripts of 3.5 and 1.5 kb were detected which hybridize to the left terminus (0.738 to 0.746 mu) of the BamHI DNA fragment B. The 1.5 kb RNA transcript was missing in the avirulent HSV-1 HFEM. Hybridization with the BssHII DNA fragment F (0.760 to 0.762 mu) led to detection of a 3.5 kb RNA transcript by HSV-1 HFEM which was missing in all other viruses tested. In contrast a 1.5 kb RNA transcript was detectable in all other virus strains with the exception of HSV-1 HFEM. The 3.5 kb transcript hybridized to the right-hand flank of the deleted region in the genome of HSV-1 HFEM (Asp718/SalI DNA fragment; 0.786 to 0.79 mu). The detection of the novel 1.5 kb RNA, which is missing in HSV-1 HFEM, and the appearance of the newly transcribed 3.5 kb RNA in HSV-1 HFEM only, indicates a new open reading frame in this particular region as a consequence of the fusion of the DNA sequences at both ends of the deletion in the genome of HSV-1 HFEM.

Chromosome Deletion↗
PubMed1988

Multiple sclerosis autoantibodies and antibodies in AIDS may deplete a brain peptide hormone.

Computerized Chou-Fasman analysis of the secondary structure of human T-cell leukemia viruses (HTLV-I, HTLV-II) and human immunodeficiency virus (HIV) envelope proteins revealed that only one antigenic epitope (amino acids EAL) is shared by the three viruses. A similar antigenic epitope is also found in human and rat brain hormone vasopressin-neurophysin. If autoantibodies in multiple sclerosis (MS) are made to the epitope EAL, they may cross-react with the envelope proteins of HTVL. It is speculated that in AIDS patients, antibodies to the antigenic epitope EAL of HIV may cross-react with brain vasopressin-neurophysin, leading to a decline in this brain peptide hormone. Thus it is hypothesized that treatment of both MS and AIDS patients with a synthetic polymer containing the amino acids EAL might eliminate the antibodies to vasopressin-neurophysin and thus alleviate some of the clinical symptoms.

Acquired Immunodeficiency Syndrome↗