Serologic reactivity using conserved envelope epitopes in feline lentivirus-infected felids.

An enzyme-linked immunosorbent assay (ELISA) based on synthetic peptides identical to lentivirus envelope protein amino acid sequences was used to study serologic reactivity of lentivirus-infected domestic cats and nondomestic felids. One feline immunodeficiency virus (FIV) peptide, P237, was consistently recognized by antibodies from FIV-infected cats, but 2 other FIV peptide antigens were not. The molecular basis for this serologic reactivity was examined.

NMR evidence for slow collective motions in cyanometmyoglobin.

Residual dipolar couplings observed in NMR spectra at very high magnetic fields have been measured to a high degree of accuracy for the paramagnetic protein cyanometmyoglobin. Deviations of these measurements from predictions based on available crystallographic and solution structures are largely systematic and well correlated within a given helix of this highly alpha-helical protein. These observations can be explained by invoking collective motion and small displacements of representative helices from their reported average positions in the solid state.

Mutations in the adrenoleukodystrophy gene.

Adrenoleukodystrophy (ALD) is a peroxisomal disorder that commonly manifests as demyelination of the central nervous system (CNS). The isolation of the ALD gene by positional cloning has led to the identification of a variety of mutations in the ALD gene. One hundred and ten mutations have been identified to date, of which approximately 50% are missense mutations.

A multiphase 113Cd NMR investigation of metalloporphyrin reorientation in cadmium-substituted myoglobin.

113Cd NMR spectroscopy in both the solution and solid state has been used to investigate the role of the metal ion and the proximal histidine on metalloporphyrin reorientation in myoglobin. Heme disorder has been known to exist for many years but understanding its mechanism has proved difficult due to the short-lived nature of the minor porphyrin isomer in native myoglobin. Cadmium-substituted myoglobin can be generated in one form which contains different insertion isomers or in another form which contains predominantly only one of these species.

Identification of war victims from mass graves in Croatia, Bosnia, and Herzegovina by use of standard forensic methods and DNA typing.

The postmortem remains of sixty-one war victims were excavated from 6 mass graves in Bosnia and Herzegovina one and a half years after interment Using standard identification methods, including the matching of medical and dental records, the recognition of distinguishing characteristics such as the use of clothing and belongings, and video superimposition, 35 persons were identified. For the remaining 26 persons identification efforts continue. DNA typing was performed at the HLA DQA1 locus and five PM system loci.

DNA-based diagnostics for adrenoleukodystrophy in a large New Zealand family.

Aim: To develop a DNA-based diagnostic test for adrenoleukodystrophy (ALD) in a large New Zealand family.

Methods: Mutation screening of the X chromosome-linked ALD gene was undertaken by direct sequencing of PCR amplified products encompassing defined exons of the ALD gene. The identification of a mutation led to the development of a simple restriction enzyme digestion protocol of a PCR amplified product to identify those individuals with the mutation.

Structure and location of the murine adrenoleukodystrophy gene.

X-linked adrenoleukodystrophy (ALD) is a degenerative neurological disease characterized by the accumulation of very long chain fatty acids in various tissues and demyelination of the central nervous system. The human gene responsible for the disease encodes a membrane-bound ATP-binding transporter protein that is located in peroxisomes. We isolated the mouse adrenoleukodystrophy gene, determined its structure, and mapped it both cytogenetically and genetically.

The sites of gene expression of atrial, brain, and C-type natriuretic peptides in mouse fetal development: temporal changes in embryos and placenta.

Atrial (ANP), brain (BNP), and C-type natriuretic peptide (CNP) belong to a family of hormones important in blood pressure and sodium homeostasis. Expression of ANP has been reported in embryo hearts, but BNP and CNP expression during development has not been described. We used in situ hybridization to identify the sites of gene expression of ANP, BNP, and CNP during development in mouse embryos at daily intervals from midgestation.

Effect of adenine methylation on the structure and dynamics of TpA steps in DNA: NMR structure determination of [d(CGAGGTTTAAACCTCG)]2 and its A9-methylated derivative at 750 MHz.

At TpA steps in DNA, the adenine base experiences exceptionally large amplitude (20 degrees-50 degrees) and slow (10 ms-1 microsecond) motion [Kennedy et al. (1993) Biochemistry 32, 8022-8035] which has been correlated with transitions between multiple conformational states [Lefevre et al. (1985) FEBS Lett.

The effects of sequence context on base dynamics at TpA steps in DNA studied by NMR.

Base dynamics, heretofore observed only at TpA steps in DNA, were investigated as a function of sequence context by NMR spectroscopy. The large amplitude conformational dynamics have been previously observed in TnAn segments where n > or = 2. In order to determine whether the dynamic characteristics occur in more general sequence contexts, we examined four self-complementary DNA sequences, [d(CTTTA-NATNTAAAG)2] (where N = A, C, T, G and N = complement of N).

Nuclear magnetic dipole interactions in field-oriented proteins: information for structure determination in solution.

The measurement of dipolar contributions to the splitting of 15N resonances of 1H-15N amide pairs in multidimensional high-field NMR spectra of field-oriented cyanometmyoglobin is reported. The splittings appear as small field-dependent perturbations of normal scalar couplings. Assignment of more than 90 resonances to specific sequential sites in the protein allows correlation of the dipolar contributions with predictions based on the known susceptibility and known structure of the protein.

Virucidal efficacy of the newer quaternary ammonium compounds.

The virucidal activity of several disinfectants containing newer generation quaternary ammonium compounds (QACs) as their active ingredients was evaluated. Disinfectants were used at the manufacturers' recommended dilutions with isolates of feline herpesvirus, feline calicivirus, and canine parvovirus, and a contact time of 10 minutes at room temperature. Detoxification of virus/disinfectant solutions was done by dialysis prior to virus assay in cell cultures.

A solid-state 2H NMR investigation of purine motion in a 12 base pair RNA duplex.

Solid-state 2H NMR spectroscopy has been used to investigate the base dynamics of a RNA oligonucleotide with a defined sequence, [r(CGCGAAUUCGCG)]2, which contains the RNA analogue of the EcoRI binding site. The C8 protons of all purines in the self-complementary dodecamer were exchanged for deuterons. The quadrupole-echo lineshapes and spin-lattice relaxation times as a function of hydration for the sample in the form of the Na salt have previously been reported.

Genomic structure, promoter sequence, and revised translation of human homeobox gene HLX1.

The human homeobox gene HLX1 appears to be involved in hemopoietic development and may represent a candidate gene for various developmental or hemopoietic disorders. We have isolated genomic clones for the gene, determined its intron-exon organization, and confirmed its map location on chromosome 1q41-q42. The transcription initiation sites of HLX1 were identified, and DNA sequences upstream of these sites were established.

Relationship of oral microflora with oral health status in Parkinson's disease.

Parkinson's disease (PD) patients report an increased craving for sweets, which may have an effect on microflora. We compared patients of PD who crave sweets with PD patients who do not. Age- and sex-matched control subjects were used, with 14 subjects in each group.

Cloning, sequencing, and in vitro expression of glycoprotein gp48 of a noncytopathogenic strain of bovine viral diarrhea virus.

Total cellular and viral RNA isolated from cells infected with noncytopathic bovine viral diarrhea virus (BVDV) strain 2724 was used for reverse transcription of viral specific sequences encoding the putative signal sequence and protein-encoding region of gp48. The cDNA template was amplified twice by the polymerase chain reaction with a nested set of primers designed from nucleotide sequences of cytopathic BVDV strains NADL and 72, and ligated into a plasmid vector. Nucleotide sequence analysis of the cloned cDNA indicated it was 921 base pairs long, encoded 307 amino acid residues, had high sequence homology to other pestiviruses, and had no significant homology to members of the Flaviviridae.

Photosensitized inactivation of infectious DNA by urocanic acid, indoleacrylic acid and rhodium complexes.

Naked, infectious single-stranded (ss) and double-stranded (ds) DNA from phages S13 and G4 were irradiated with 308 nm UV radiation in the absence and presence of several photobiologically active compounds: E- and Z-urocanic acid (E- and Z-UA), their methyl esters (E- and Z-MU), E- and Z-indoleacrylic acid (E- and Z-IA), cis-dichloro-bis(1,10-phenanthroline)rhodium(III) chloride (cDCBPR) and tris(1,10-phenanthroline)rhodium (III) perchlorate (TPR). E-urocanic acid protects against cyclobutane pyrimidine dimer formation in ssDNA but concomitantly photosensitizes the formation of other lesions that inactivate ssDNA. Z-urocanic acid also protects ssDNA against such dimerization but without the associated sensitized damage.

DNA polymerase II of Escherichia coli in the bypass of abasic sites in vivo.

The function of DNA polymerase II of Escherichia coli is an old question. Any phenotypic character that Pol II may confer upon the cell has escaped detection since the polymerase was discovered 24 yr ago. Although it has been shown that Pol II enables DNA synthesis to proceed past abasic sites in vitro, no role is known for it in the bypass of those lesions in vivo.

Unusual kinetics of uracil formation in single and double-stranded DNA by deamination of cytosine in cyclobutane pyrimidine dimers.

Mutagenesis studies have indicated that the deamination of cytosine in UV-induced cyclobutane pyrimidine dimers is a key part of an error-free process that can account for most of the C-->T base specificity that frequently characterizes UV-induced mutagenesis. The kinetics of deamination, as inferred from the mutagenic effect of delayed photoreactivation, is remarkable in its resemblance to a step function. To study the kinetics from a different point of view, we used an enzymatic approach combining photolyase and uracil-N-glycosylase treatment to detect the formation of uracil in UV-irradiated single or double-stranded infectious DNA of phage S13.

Cloning and sequencing of a t(14;19) breakpoint that involves the C mu switch region.

The t(14;19) is a recurring translocation found in a small number of cases of chronic B-cell leukemia (CLL). We have cloned and sequenced the breakpoint in a patient with a t(14;19) and shown that the breakpoint on chromosome 14 occurred in the C mu switch region, and that the breakpoint on chromosome 19 occurred in the 5' untranslated region of the BCL3 gene. This is in contrast to all the other reported cases with a t(14;19) in which the breakpoints on chromosome 14 occurred in the C alpha 1 or C alpha 2 switch region, and the breakpoints on chromosome 19 occurred upstream of the BCL3 gene.

Mobility at the TpA cleavage site in the T3A3-containing AhaIII and PmeI restriction sequences.

Lefevre et al. originally observed conformational transitions at the TpA step in the TTAA Pribnow box sequence of the trp promoter [Lefevre, J.-F.