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At least 379 records · Page 21Linked to original sources
PubMed1988

Content and organization of the human Ig VH locus: definition of three new VH families and linkage to the Ig CH locus.

We present a detailed analysis of the content and organization of the human immunoglobulin VH locus. Human VH genes representing five distinct families were isolated, including novel members belonging to two out of three of the known VH gene families (VH1 and VH3) as well as members of three new families (VH4, VH5, and VH6). We report the nucleotide sequence of 21 novel human VH genes, many of which belong to the three new VH gene families. In addition, we provide a preliminary analysis of the organization of these gene segments over the full extent of the locus. We find that the five multi-segment families (VH1-5) have members interspersed over nearly the full 1500-2000 kb of the VH locus, and estimate that the entire heavy chain locus covers 2500 kb or less. Finally, we provide the first report of the physical linkage of the variable and constant loci of a human Ig gene family by demonstrating that the most proximal known human VH segments lie within 100 kb of the constant region locus.

Animals↗
PubMed1988

Effects of nitrosation on the chemical composition and epidermal carcinogenicity of the nitrogen-rich fraction of a high-boiling coal liquid.

An 800-850 degrees F solvent-refined coal-II liquid was fractionated into chemical classes to obtain the aliphatic hydrocarbons, polycyclic aromatic hydrocarbons (PAH), nitrogen-containing polycyclic aromatic compounds (NPAC), and hydroxy-substituted PAH (hydroxy-PAH). The isolated NPAC fraction was refractionated by chemical class both before and after undergoing a nitrosation reaction. The nitrosated and non-nitrosated refractionated NPAC fractions were further subfractionated into secondary amine (pyrroles), primary amine-enriched (amino-PAH), and tertiary amine (azaarene) classes. The PAH and hydroxy-PAH composition of the NPAC fraction increased upon nitrosation, whereas the amino-PAH fraction composition decreased upon nitrosation. Nitrosation of standards indicated the amino-PAH compounds reacted to form parent PAH, chloro-substituted PAH, and methoxy-substituted PAH when analyzed by high-resolution gas chromatography (GC) and GC/mass spectrometry (MS). Some easily oxidized PAH compounds reacted to form ketones and quinones. All other standard reference compounds, chosen to be representative of the major chemical classes of compounds present in coal liquefaction materials, were unchanged by the nitrosation reaction. The amino-PAH of the nitrosated NPAC fraction reacted to form parent and some chloro-substituted PAH when analyzed by low-voltage direct-probe MS in addition to the methods given above. Epidermal carcinogenesis studies with the PAH, NPAC, nitrosated NPAC, and hydroxy-PAH fractions isolated from the 800-850 degrees F coal liquid indicated the PAH and NPAC were the most important determiners of skin carcinogenesis, with the PAH giving a higher response than the NPAC. The tumorigenicity of the NPAC was drastically reduced by nitrosation, probably due to the destruction of the amino-PAH upon nitrosation.

Animals↗
PubMed2002

Growth arrest in G1 protects against oxygen-induced DNA damage and cell death.

Although oxygen is required for normal aerobic respiration, hyperoxia (95% O(2)/5% CO(2)) damages DNA, inhibits proliferation in G1, S and G2 phases of the cell cycle, and induces necrosis. The current study examines whether growth arrest in G1 protects pulmonary epithelial cells from oxidative DNA damage and cell death. Mv1Lu pulmonary adenocarcinoma cells were chosen for studies because hyperoxia inhibits their proliferation in S and G2 phase, while they can be induced to arrest in G1 by altering culture conditions. Hyperoxia inhibited proliferation, increased intracellular redox, and rapidly reduced clonogenic survival. In contrast, Mv1Lu cells treated with transforming growth factor (TGF)-beta1, deprived of serum or grown to confluency, arrested and remained predominantly in G1 even during exposure. Growth arrest in G1 significantly enhanced clonogenic survival by 10-50-fold. Enhanced survival was not due to reduction in the intracellular redox-state of the cells, but instead was associated with reduced DNA strand breaks and p53 expression. Our findings suggest that the protective effects of G1 is mediated not simply by a reduction in intracellular ROS, but rather through an enhanced ability to limit or rapidly recognize and repair damaged DNA.

Adenocarcinoma↗
PubMed1995

Ligation of CD40 on fibroblasts induces CD54 (ICAM-1) and CD106 (VCAM-1) up-regulation and IL-6 production and proliferation.

CD40 was originally described as a functionally significant B cell surface molecule. However, CD40 is also expressed on monocytes, dendritic cells, epithelial cells, and basophils. We now report that synovial membrane (SM) or dermal fibroblasts also express cell surface CD40 in vitro. Fibroblast CD40 expression declines with increasing time in culture and recombinant interferon-gamma (rINF-gamma) induces fibroblast CD40 up-regulation. This effect of rINF-gamma is augmented by recombinant interleukin-1 alpha or recombinant tumor necrosis factor-alpha. CD40 expression on fibroblasts is functionally significant because CD40L-CD40 interactions induce SM fibroblast CD54 (intercellular adhesion molecule-1) and CD106 (vascular cell adhesion molecule-1) up-regulation. Moreover, ligation of CD40 augments IL-6 production by SM fibroblasts and induces fibroblasts to proliferate. In addition, rINF-gamma enhances the effect of CD40L-CD40 interactions on fibroblast proliferation. Taken together, these studies show that fibroblasts can express CD40, cytokines can regulate fibroblast CD40 expression, and CD40 ligation induces fibroblast activation and proliferation.

Animals↗
PubMed1991

An evaluation of three loading configurations for the in vitro testing of femoral strains in total hip arthroplasty.

Despite recent advances in total hip arthroplasty, proximal femoral resorption and osteopenia remain problems. To analyze the proximal strain effects of three different loading conditions, strains produced in intact and postarthroplasty femora have been compared. Ten adult cadaveric femora of similar size, shape, and rigidity were tested. Ten strain-gauge rosettes were positioned on each femur. To simulate the "single leg support" phase of gait, fixtures were developed to load the femora under three different configuration: the VS (vertical shaft) configuration, with a vertically orientated femur having rotational freedom proximally and distally; the ITB (iliotibial band) configuration of a femur with rotational freedom positioned 11 degrees from the vertical, with a strain-gauge adjustable metallic simulation of the lateral muscles of the thigh; and the ABD (abductor) configuration of a femur with rotational freedom positioned 11 degrees from the vertical, with a strain-gauge adjustable metallic simulation of the abductor muscles. Each femur was loaded less than or equal to 600 N through the medial point, located at one sixth of the transcondylar distance. Strain patterns and magnitudes produced by the three loading configurations were quite different in both the intact and postarthroplasty femora. Both the ITB and the ABD configurations resulted in greater proximal medial compression and lateral tension than did the VS configuration. The magnitudes of the proximal strains were significantly greater in the ABD configuration (p less than 0.05). Postarthroplasty femora showed similar proximal results. It is proposed that meaningful strain data for the physiologically loaded femur can be obtained only with simulations that include the forces produced by the iliotibial band. To overcome the indeterminate nature and biological variation in these forces, the studies have to consider a range of forces.

Femur↗
PubMed2005

Determination of chemical agent optical clearing potential using in vitro human skin.

BACKGROUND AND OBJECTIVES: Optical clearing is a method that temporarily reduces the optical scattering of biological tissues. STUDY DESIGN/MATERIALS AND METHODS: To determine the optical clearing potential (OCP) of various chemical agents, we examined the change in reduced scattering coefficients of in vitro human skin after application directly to the dermal side. RESULTS/CONCLUSIONS: We found that the mean agent OCP did not correlate with refractive index or osmolality. Lipophilic hyperosmotic agents had a mean OCP less than unity, indicating increased optical scattering.

Culture Techniques↗
PubMed2003

Role of beta-adrenoceptor subtypes in mediating relaxation of the pig bladder trigonal muscle in vitro.

AIMS: To investigate the role of beta-adrenoceptor subtypes in mediating relaxation of the pig bladder trigonal muscle in vitro. MATERIALS AND METHODS: Longitudinal strips of trigonal muscle were isolated, and the mucosa and serosa removed. Tissues were precontracted with KCl, and beta-adrenoceptor agonists (isoprenaline or salbutamol) were added cumulatively, and concentration-relaxation curves (CRCs) were obtained. CRCs to agonists were obtained in the absence and presence of antagonists and antagonist affinity values were calculated. RESULTS: Isoprenaline (non-selective beta-agonist) relaxed with high potency (pEC(50) = 7.2). Propranolol antagonized CRCs to isoprenaline with a high affinity (apparent pK(B) = 8.8), but the Schild plot had a slope significantly (P < 0.01) less than unity (0.61), suggesting that responses were mediated by more than one beta-adrenoceptor subtype. CGP20712A (beta(1)-antagonist) antagonized responses to isoprenaline with a low affinity (apparent pK(B) = 5.13), indicating beta(1)-adrenoceptors did not participate in the response. The affinity of ICI118551 (beta(2)-antagonist) for antagonism of responses to isoprenaline was also relatively low (apparent pK(B) = 6.9), and the Schild slopes were significantly (P < 0.01) less than unity (0.58). SR59230A (beta(3)-antagonist) antagonized CRCs to isoprenaline with a relatively low affinity (apparent pK(B) = 7.5), and with a Schild slope significantly (P < 0.01) less than unity (0.86), indicating that responses may be mediated by more than one beta-adrenoceptor subtype. In contrast to that observed with isoprenaline, the beta(2)-adrenoceptor selective agonist salbutamol induced relaxation with a relatively low potency (pEC(50) = 6.6) and with maximum responses of 80% of that to isoprenaline. ICI118551 competitively antagonized (Schild plot of unity) responses to salbutamol with a high affinity (pA2 = 8.3). CONCLUSIONS: These data suggest that beta-adrenoceptor mediated responses to isoprenaline of the bladder trigone are mediated via both the beta(2)- and beta(3)-adrenoceptor subtypes whilst responses to salbutamol appear to be mediated only via the beta(2)-adrenoceptor.

Adrenergic beta-Agonists↗
PubMed2000

Sequence determination and analysis of the full-length genome of colorado tick fever virus, the type species of genus Coltivirus (Family Reoviridae).

The Colorado tick fever virus (CTFV) is the type species of genus Coltivirus, family Reoviridae. Its genome consisting of 12 segments of dsRNA was completely sequenced. It was found to be 29,174 nucleotides long (the longest of all Reoviridae genomes characterized to date). Conserved sequences at the 5' end (SACUUUUGY) and at the 3' end (WUGCAGUS) of the 12 segments were identified. The analysis of the putative proteins deduced from the nucleotide sequences permitted to identify functional motifs. In particular, the VP1 was identified unambiguously as the viral RNA dependent RNA pylmerase (RDRP) (VP1pol), with a GDD located at a similar position to Reoviridae RDRPs. In other genes, RGD cell-binding, NTPAse, single strand binding protein and kinase motifs were identified. Comparison with Reoviridae proteins showed significant similarities to RDRPs (CTFV-VP1) and sigma C protein of orthoreovirus (CTFV-VP6). Similarities to nonviral enzymatic proteins, such as methyltransferases, NTPAses, RNA replication factors, were also identified.

Animals↗
PubMed2001

Complete coding sequence of the Alkhurma virus, a tick-borne flavivirus causing severe hemorrhagic fever in humans in Saudi Arabia.

To date, tick-borne flaviviruses responsible for hemorrhagic fever in humans have been isolated in Siberia (Omsk hemorrhagic fever virus), India (Kyasanur Forest disease virus, KFDV), and in Saudi Arabia (Alkhurma virus, ALKV). Prior to this study, only partial coding sequences of these severe pathogens had been determined. We report here the complete coding sequence of ALK virus, which was determined to be 10,248 nucleotides (nt) long, and to encode a single 3,416 amino acid polyprotein. Independent analyses of the complete polyprotein and the envelope protein provided genetic and phylogenetic evidence that ALKV belongs to the tick-borne flavivirus group, within which it is most closely related to KFDV. Analysis of structural genes, genetic distances, and evolutionary relationship indicate that ALKV and KFDV derived from a common phylogenetic ancestor and constitute two genetic subtypes of the same virus species according to current genetic criteria of classification.

Flavivirus↗
PubMed2002

Lymphocytes expressing alpha1beta1 integrin (very late antigen-1) in peripheral blood of patients with arthritis are a subset of CD45RO(+) T-cells primed for rapid adhesion to collagen IV.

We report that very late antigen-1 (VLA-1(+)) CD3(+)CD45RO(+) T-cells are selectively segregated from VLA-1(-) peripheral blood (PB) mononuclear cells (MC), in which CD3(+) T-cells are evenly CD45RO(+) and CD45RO(-), when PBMC are stained with a monoclonal antibody (mAb) to VLA-1 and passaged on immunomagnetic columns. In contrast, both VLA-1(+) and VLA-1(-) MC isolated from synovial fluid (SF) are mainly CD45RO(+)CD3(+) T-cells. VLA-1(+) MC formed 13 +/- 5.3% of MC eluting from columns loaded with PBMC of patients with seropositive rheumatoid arthritis (n = 6) and 2.3 +/- 1.6% of patients (n = 4) with other arthritides (P < 0.022). Importantly, only the VLA-1(+) MC from PB and SF adhered to collagen IV upon triggering with phorbol 12-myristate 13-acetate. Moreover, adhesion and migration on collagen IV were preferentially maintained in lines cultured from VLA-1(+) T-cells, and both were inhibited by mAb to the VLA-1 alpha1 I domain. These results suggest that VLA-1(+) CD45RO(+) T-cells in patients with arthritis could play a role in both systemic and local inflammation by rapidly adhering to collagen IV.

Antibodies, Monoclonal↗
PubMed1999

Identification of candidate Drosophila olfactory receptors from genomic DNA sequence.

We have taken advantage of the availability of a large amount of Drosophila genomic DNA sequence in the Berkeley Drosophila Genome Project database ( approximately 1/5 of the genome) to identify a family of novel seven transmembrane domain encoding genes that are putative Drosophila olfactory receptors. Members of the family are expressed in distinct subsets of olfactory neurons, and certain family members are restricted to distinct portions of the olfactory system. This pattern of expression has interesting similarities to and differences from the expression patterns observed for olfactory receptors in vertebrates. The Drosophila olfactory system is simpler than mammalian systems, yet it is complex enough to present a fascinating system in which to study neural information processing. Moreover, the powerful genetic manipulations available in Drosophila, when combined with electrophysiological and behavioral analyses, make this an attractive model system in which to study olfactory discrimination.

Amino Acid Sequence↗
PubMed1984

Construction of human T-cell hybrids with helper function.

Human T-cell hybrids with helper activity were obtained after fusion of phytohemagglutinin-activated normal human T cells with a 6-thioguanine-resistant, aminopterin-sensitive human T-cell line. This mutant line, designated CEM-T15, was derived from the human T-cell line CEM after mutagenesis with ethyl methanesulfonate. The polyethylene glycol induced fusion and the selection in hypoxanthine- aminopeterin -thymidine medium were performed by modification of standard somatic cell hybridization techniques. After fusion, the strategy for selecting hybrids consisted in screening growing cultures for the presence of cells expressing the OKT3 cell surface differentiation antigen. OKT3 was chosen because it is present in 85-95% of normal human T cells but absent from CEM-T15 cells. Thus, OKT3+ cells growing 5-7 weeks after fusion most likely represented hybrids between normal T cells (OKT3+) and continuously growing CEM-T15 cells (OKT3-). Several of the hybrids were tested for their capacity to promote pokeweed mitogen-induced antibody production by B cells. These experiments demonstrated that many of the hybrids had helper activity. Periodical testing of these uncloned hybrids for helper activity revealed functional instability, with most of the hybrids losing helper activity after 20 weeks of continuous culture. However, early and repeated cloning of the same hybrids resulted in a series of hybrid clones with helper activity still present more than 8 months after fusion. In more recent fusions, we have demonstrated that human helper hybrids producing helper factor(s) can also be obtained. These and similar hybrids with different functions will be of considerable importance in further studies of the immunobiology of human T lymphocytes.

Antigens, Surface↗
PubMed1987

Cardiac alpha- and beta-adrenoceptor sensitivity and binding characteristics after chronic reserpine pretreatment.

Cardiac alpha- and beta-adrenoceptor sensitivities were examined after chronic pretreatment of rats with reserpine. Increases in sensitivity would indicate that the receptor is under the influence of the sympathetic innervation, removal by catecholamine depletion with reserpine of the tonic effect of neurotransmitter release would permit receptor upregulation. The positive inotropic responses of paced left atria and papillary muscles and the positive chronotropic responses of spontaneously beating right atria were recorded. A concentration-response curve to isoprenaline (beta-adrenoceptor-mediated) was followed, in the presence of beta-blockade, by one to methoxamine (alpha-adrenoceptor-mediated). Methoxamine exerted positive inotropy of left atria and papillary muscles, the maxima being 43.2 +/- 2.7 and 26.8 +/- 4.4% of the isoprenaline maxima. A small positive chronotropy (16.5 +/- 5.6% maximum) of right atria occurred. After pretreatment with reserpine (1.0 mg kg-1 i.p. daily) for 7 days, the three preparations displayed supersensitivity to isoprenaline, revealed as a significant displacement (P less than 0.05) of the concentration-response curves to the left of those for control rats. Reserpine pretreatment, however, had no effect on the sensitivity to methoxamine. The increase in beta-adrenoceptor sensitivity to isoprenaline after reserpine pretreatment was accompanied by a significant 41.3% increase (P less than 0.05) in the number of [3H]-dihydroalprenolol [( 3H]-DHA) binding sites (Bmax) in ventricular membranes, although the dissociation constant (KD) was unaffected. There were more alpha-adrenoceptor [3H]-prazosin binding sites in ventricular than atrial membranes. However, there was no difference in KD or Bmax between reserpine-pretreated and control tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Cardiac postjunctional supersensitivity to beta-agonists after chronic chemical sympathectomy with 6-hydroxydopamine.

The sensitivity to sympathomimetic amines of isolated atria removed from sham-injected and 6-hydroxydopamine-treated (6-OHDA) guinea-pigs was examined in the presence of an extraneuronal uptake blocker and an alpha-adrenoceptor antagonist. Three weeks of pretreatment with 6-OHDA resulted in leftwards shifts of the dose-response curves for the positive chronotropic and inotropic responses of right and left atria to isoprenaline. The responses to the partial agonist salbutamol were also potentiated after 6-OHDA pretreatment, revealed as an increase in the maximum response relative to isoprenaline. The supersensitivity was post-synaptic in origin and independent of changes in disposition or metabolism, since it was observed with agonists immune to neuronal uptake and O-methylation, and in the presence of extraneuronal uptake inhibition by metanephrine. It was also specific for the beta-adrenoceptor, no supersensitivity to histamine being found. In the right atria, the supersensitivity was partially masked by an opposing depressant effect after 6-OHDA pretreatment which was observed with histamine. Dissociation constants (KA) for the left atrial inotropic responses to orciprenaline were determined by use of the antagonist Ro 03-7894. Atria from 6-OHDA-pretreated animals were supersensitive to orciprenaline, but the KA value did not differ from that after sham injection. It could therefore be concluded that the increase in sensitivity was not due to an increase in affinity for the beta-adrenoceptor.

Adrenergic beta-Agonists↗