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Biomedical subjects

R Anand

Publications and source records attributed to R Anand.

At least 199 records · Page 11Linked to original sources

Communicating oesophageal duplication: a report of two cases.

Alimentary duplications which include a variety of alimentary cysts, diverticula and tubular malformations are rare abnormalities. Most of them, especially in relation to the oesophagus, do not communicate with the lumen. We present two very rare cases of communicating oesophageal duplication. Both are left sided and unassociated with any other congenital abnormality. These features again are uncommon with such duplications.

Esophagus↗

Molecular characterization of a beta zero-thalassemia resulting from a 1.4 kilobase deletion.

We report the characterization of a beta zero-thalassemia in an American Black with unusually high HbA2 and HbF levels. Genomic southern analysis indicated that the individual was heterozygous for a deletion that began within the second intervening sequence of the beta-globin gene and extended approximately 1.4 kb in the 5' direction. A clone spanning the breakpoint on the abnormal chromosome was isolated and further mapped, and the deletion joint was sequenced. Comparison of the normal beta-globin gene and its 5' flanking region with the deletion joint sequence indicated that the 5' breakpoint for this deletion was 484 base pairs (bp) 5' to the transcriptional start site for the beta-globin gene and the 3' breakpoint was 908 bp into the beta-globin gene; the deletion removed a total of 1,393 bp. Comparison of the normal 5' and 3' breakpoint sequences indicated that this deletion was the result of a "clean" nonhomologous breakage and reunion event; ie, no spurious bases were added during the recombinational event. Analysis of the breakpoints of this deletion together with the breakpoints of two other small deletions involving the beta-globin gene suggests that the breakpoints may occur at DNA polymerase alpha pause sites.

Base Sequence↗

Iniencephaly.

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Abnormalities, Multiple↗

A model for the separation of large DNA molecules by crossed field gel electrophoresis.

The idea that large DNA molecules adopt a stretched conformation as they pass through gels suggests a simple mechanism for the separation of DNA by crossed field electrophoresis: at each change in field direction a DNA molecule takes off in the new direction of the field by a movement which is led by what was formerly its back end. The effect of this ratcheting motion is to subtract from the DNA molecule's forward movement, at each step, an amount which is proportional to its length. We find that this model explains most of the features of the separation, and we describe experiments, using a novel electrophoresis apparatus, which support the model. The apparatus turns the gel between two preset orientations in a uniform electric field at preset time intervals. This separation method has the practical advantage over some others that the DNA molecules follow straight tracks. A further advantage is that the parameters which determine the separation are readily predicted from the simple theory describing their motion.

DNA↗

Non-cytocidal natural variants of human immunodeficiency virus isolated from AIDS patients with neurological disorders.

To understand the mechanism of HIV-mediated neuropathology five viral isolates were obtained from four AIDS cases with central nervous system manifestations as the primary involvement. The isolates were identified as HIVs by antigenic cross-reactivity and nucleic acid hybridizations to HIV-specific antibodies and DNA probes. The replication and cytopathic properties of these isolates were studied and compared with lymphadenopathy-associated virus (HIVLAV). All isolates had replication competence equivalent to LAV, but four isolates did not kill T4 (CD-4) cells. This isolation of non-cytocidal natural variants of HIV raises the possibility that in some AIDS cases the neurological disorders might be due to HIV variants that are non-cytocidal to T4 cells. The results also indicate that virus replication and cytotoxicity are not always concordant functions in HIV.

Acquired Immunodeficiency Syndrome↗

Identification and characterisation of proteins associated with the rhoptry organelles of Plasmodium falciparum merozoites.

We describe antigens of Plasmodium falciparum recognised by murine monoclonal antibodies which by immunofluorescence react with the rhoptry organelles of the extracellular merozoite stage. Immunoblotting shows that the antibodies recognise two major parasite antigens of Mr 82 and 65 kilodaltons (kDa). Immunoprecipitations from detergent extracts of [35S]-methionine-labelled parasites show that the 82-kDa and 65-kDa antigens are parasite proteins. Pulse-chase experiments on synchronous parasite cultures show that the 82-kDa protein is synthesised during early schizogony and is later processed into the 65-kDa antigen in segmenting schizonts. In Nonidet P-40, these antigens are non-covalently associated with two other proteins of 40 kDa and 42 kDa. The 40/42-kDa doublet is synthesised in parallel with the 82 kDa antigen and persists, apparently unchanged, till the end of the cell cycle.

Animals↗

Infection and transformation of Fv-2rr erythroprogenitor cells with Friend virus.

Friend virus was used to infect and transform Fv-2rr erythroprogenitor cells in vivo. The RB (Fv-2rr) cell line was characteristic of Friend virus-induced cell lines in Fv-2ss mice, i.e., it produced infectious Friend virus and synthesized hemoglogin. The RB (Fv-2rr) cell line expressed the envelope protein of the spleen focus-forming virus (gp52) and a novel, related envelope protein (gp48). The results demonstrate that Fv-2rr erythroprogenitor cells can be infected and transformed in vivo.

Animals↗

Characterization of cell lines derived from enlarged spleens induced in C57BL/6 mice by the variant BSB strain of Friend erythroleukemia virus.

BSB is a variant strain of Friend virus selected for pathogenicity in C57BL/6 mice that are resistant to parental Friend virus strains by virtue of their homozygosity for the recessive Fv-2r allele (Steeves et al., 1970, Int. J. Cancer 5, 346-356). Lines and clones of erythroleukemia cells could readily be established in culture from the enlarged spleens of BSB-infected Fv-2r homozygotes. All lines expressed viral gene products and could be induced to express hemoglobin. Some lines produced infectious virus. In addition to the viral envelope-related proteins (gPr85, gp70, and gp52) detected by precipitation with goat anti-Rauscher gp70 antiserum from tumor cell lines induced by parental Friend virus strains, BSB-induced cell lines also expressed gp80, p52, and gp45 products precipitable with the same antiserum. A rat monoclonal antibody that recognizes an epitope of an amino-terminal region of gp52 (Wolff et al., 1982, J. Virol. 43, 472-481) also precipitated the gp80 and gp45 viral proteins. The data indicate that the BSB strain of Friend virus is oncogenic in Fv-2r homozygotes. Transformation is correlated with the expression of an altered SFFV env-gene product.

Animals↗

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↗

Topography of epitopes on a polymorphic schizont antigen of Plasmodium falciparum determined by the binding of monoclonal antibodies in a two-site radioimmunoassay.

The topographic distribution of common and variant epitopes on two divergent allelic forms of the 185-205K schizont glycoprotein of Plasmodium falciparum were studied by a two-site radioimmunoassay using monoclonal antibodies. Similarities in the conformation of the two molecules were apparent. On both antigens two distinct regions were mapped, each comprising of both strain-common and polymorphic epitopes. Epitopes common to the two PSAs were found to be closely associated with different variable epitopes in tertiary structure. It is suggested that this may contribute to parasite evasion of the host immune response.

Animals↗

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↗

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↗

DNA and protein heterogeneity in serial isolates of human immunodeficiency virus (HIV-1): indication of change in vivo.

Structural heterogeneity in human immunodeficiency virus (HIV-1) isolates from different sources has been reported. In order to investigate if the virus exhibits heterogeneity in vivo, HIV-1 was isolated over a 3 year period (1983-1985) from a blood donor who progressed from the asymptomatic carrier state to frank AIDS. The HIV-1 specific proteins were characterized and compared by metabolic labelling and immunoprecipitation of infected cells and by Western blot analysis of gradient purified isolates. The data indicate polymorphism in the apparent size of gag gene-encoded p55 and env gene-encoded p41 proteins. The possibility of genomic variations was evaluated by studying the restriction enzyme polymorphism of integrated proviral DNA. Genetic diversity between 1983, 1984 and 1985 isolates was demonstrated by a change in the number and/or size of restriction fragments. In addition, genetic variation was also observed in HIV-1 isolated from the blood transfusion recipient of this donor. Subsequent analysis of the quantitative and qualitative titre of HIV-1 specific antibodies in the donor recipient sera demonstrated a concomitant change with the observed structural variation in the virus. These studies may be useful in elucidating virus or host-related critical events which lead to the onset of AIDS following infection with HIV-1.

Antibodies, Monoclonal↗