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Biomedical subjects

L Hall

Publications and source records attributed to L Hall.

At least 19 recordsLinked to original sources

Identification of an abundant monkey epididymal transcript encoding a homologue of human CAMPATH-1 antigen precursor.

A number of cDNA clones encoding a small (0.5-0.6 kb) transcript have been isolated from a monkey (Macaca fascicularis) epididymal cDNA library. DNA sequence analysis indicates that this abundant epididymal transcript is homologous to the human CAMPATH-1 (CDw52) antigen precursor, a GPI-anchored membrane glycoprotein previously described on lymphocytes and monocytes.

Amino Acid Sequence

A mammalian epididymal protein with remarkable sequence similarity to snake venom haemorrhagic peptides.

Following spermatogenesis in the testis, mammalian spermatozoa pass into the epididymis, where they undergo changes which confer on them forward motility and the ability to recognize and penetrate the egg. Many of these maturation events involve androgen-regulated epididymal proteins which become associated with the sperm membrane, and/or effect changes to integral sperm membrane proteins. Here we report the sequence of an 89 kDa androgen-regulated protein from rat (Rattus norvegicus) and monkey (Macaca fascicularis) epididymis that is synthesized exclusively in the caput region and is localized on the apical surface of its principal epithelial cells. This protein shows remarkable similarity to a variety of proteases and disintegrins found in snake venoms and is similar to, but distinct from, the guinea-pig sperm surface PH-30 alpha/beta complex recently implicated in sperm-egg recognition and fusion.

ADAM Proteins

Genetic evidence for an androgen-regulated epididymal secretory glutathione peroxidase whose transcript does not contain a selenocysteine codon.

Epididymal glutathione peroxidase (GPX) has been suggested as a major factor in combating loss of fertility of spermatozoa due to lipid peroxidation. We report here the isolation and sequence of putative GPX cDNAs from rat (Rattus rattus) and cynomolgus-monkey (Macaca fascicularis) epididymis, which exhibit marked sequence identity with known GPXs. In both species the cDNAs encode predicted preproteins containing 221 amino acid residues. Unlike other characterized GPX sequences, epididymal GPX mRNA does not contain a selenocysteine codon (UGA). However, sequence comparison and molecular-modelling studies suggest a high degree of structural conservation between epididymal and other GPXs. Transcripts corresponding to epididymal GPX are not detected in a variety of other tissues (liver, spleen, kidney and testis) and appear to be androgen-regulated in the epididymis.

Amino Acid Sequence

Rod opsin cDNA sequence from the sand goby (Pomatoschistus minutus) compared with those of other vertebrates.

The absorbance spectra of rods from the sand goby were measured by using microspectrophotometry. Analysis of the averaged spectra shows that the rod visual pigment has a maximum absorbance (lambda max) at approximately 501 nm. A sand goby retinal cDNA library was constructed and then screened with a partial sand goby rod opsin clone obtained by the polymerase chain reaction (PCR). The screening of the library yielded a full length rod opsin clone. The cDNA sequence and deduced amino acid sequence of this clone are compared with those of other vertebrate rod opsins.

Amino Acid Sequence

Structure and expression of the rat epididymal secretory protein I gene. An androgen-regulated member of the lipocalin superfamily with a rare splice donor site.

The complete rat epididymal secretory protein I (ESP I) gene was isolated from a genomic library constructed in bacteriophage lambda Charon 4A. The complete nucleotide sequence of the gene and its immediate 5' and 3' flanking sequences were determined. Interesting features include the presence of a rare, but functional, splice donor site (...GC) and the presence of a putative androgen-receptor-binding element. A detailed analysis of ESP I regulation was carried out after castration and subsequent testosterone treatment, demonstrating the requirement for androgens. Efferent-duct ligation and cryptorchism, on the other hand, had no effect on the steady-state concentrations of ESP I transcripts. Comparison of the exon/intron organization of the ESP I gene with those of members of the lipocalin superfamily provides strong support for a common ancestral origin.

Amino Acid Sequence

Effect of reciprocal complementation of two defective human immunodeficiency virus type 1 (HIV-1) molecular clones on HIV-1 cell tropism and virulence.

Human immunodeficiency virus type 1 (HIV-1) displays both interstrain and intrastrain genetic variability. Virus populations with extensive microheterogeneity have been defined as swarms or quasispecies. Many of the genomes within HIV-1 swarms appear to be defective in one or more genes required for viral replication. It is unclear to what extent defective viruses play a role in the process of HIV-1 infection or in the pathogenesis of AIDS. We have isolated two biologically active HIV-1 clones: LW 12.3, which contains defects in the vif and vpr genes, and MN ST.1, which has a defect in the vpu gene. LW 12.3 is unable to replicate in peripheral blood mononuclear cells (PBMC). The growth of MN-ST.1 in SupT1 cells is marked by a 3-week lag in extracellular virus production and by the presence of unusually abundant viral buds. We demonstrate here that coinfection of PBMC with these two partially defective HIV-1 clones extends the cellular host range of LW 12.3, significantly increases the replication rate of both viral genomes, and eliminates the delay in production observed with the vpu-defective MN ST.1. When the lesions in vpr and vif of LW 12.3 are repaired, the resultant virus grows normally in PBMC. This is also the case when only vif is repaired, indicating that complementation of LW 12.3 in PBMC by MN ST.1 is mediated by vif in trans. The reciprocal complementation results in a dramatic increase of HIV-1 virulence. This two-component model represents a simplified version of the in vivo situation and illustrates one way in which interaction of defective viruses could increase the spread of infection and progression of disease.

Base Sequence

Use of coculture of colonic mucosal biopsies to investigate the release of eicosanoids by inflamed and uninflamed mucosa from patients with inflammatory bowel disease.

Eicosanoid production was measured in cultured biopsies of colonic mucosa from control patients, with the irritable bowel syndrome, and from patients with proctosigmoiditis and with colonic Crohn's disease. Cultured inflamed colonic mucosa from patients with proctosigmoiditis and Crohn's disease produced more prostaglandin E2 and leukotrienes C4 than control tissues. In addition, eicosanoid production by macroscopically uninflamed or 'quiescent' mucosa from the right colon was examined in patients with proctosigmoiditis and between skip lesions in Crohn's disease patients. In the proctosigmoiditis group quiescent mucosa produced eicosanoids in similar quantities to control tissue. Coculture of quiescent plus inflamed tissue however, generated a marked increase in eicosanoid output in 12 of 20 of the patients and this was similar to the quantity obtained from two pieces of inflamed tissue. In the Crohn's disease group, quiescent mucosa produced more eicosanoids than control mucosa but production was markedly stimulated by coculture with inflamed mucosa in all patients. These findings suggest that in some patients with proctosigmoiditis and in all patients with Crohn's disease quiescent mucosa appears to be sensitised. A small but significant increase in the macrophage population may be partly responsible but it is likely that these and other cells are primed to release eicosanoids, and may be induced to do so by soluble mediators produced by actively inflamed tissue.

Adult

Venous stenoses in patients who undergo hemodialysis: treatment with self-expandable endovascular stents.

During a 32-month period, 25 modified self-expandable endovascular stents were placed in 20 hemodialysis access sites to treat 21 stenoses and four occlusions in 19 patients. All stenoses were initially dilated with a high-pressure balloon. The initial success rate was 90% (18 of 20 access sites). The stents were patent from 0 to 960 days (mean duration of follow-up, 309 days). At 2 years follow-up, the patency rates were as follows: primary, 25%; secondary, 34%; and tertiary, 42%. The morbidity rate associated with this procedure was 15% (three of 20 sites); the mortality rate, 5% (one of 19 patients). All five complications (graft or native-vein thrombosis [n = 2], nerve deficits [n = 2], and death secondary to sepsis [n = 1]) occurred in three patients early in the study, before the use of prophylactic antibiotics and refinements in technique. The best results occurred in patients in whom the stenosis or occlusion affected a large vein that had no acute angle and was away from venous confluences such as the femoral and iliac regions.

Constriction, Pathologic

Clinical reasoning process for service provision in the public school.

This paper outlines the clinical reasoning process used to guide decisions on the provision of occupational therapy services in the Wake County Public School System in North Carolina. The process is based on a theoretical framework derived from occupational therapy theory and public law. Benefits of using the clinical reasoning process include (a) increased consistency of decision making among therapists; (b) increased appropriateness of decisions regarding whether a student needs educationally based occupational therapy services, what type of occupational therapy service would meet the student's need, and how often this service should be provided; and (c) improved ability of therapists to articulate to all those involved with a student the reasoning behind decisions to provide educationally based occupational therapy services. The schematic diagrams that depict this process provide a useful tool for therapists with varied work experiences entering school-based practice.

Child

Sequence and expression of the gene encoding 3-phosphoglycerate kinase from Bacillus stearothermophilus.

The structural gene (pgk) encoding 3-phosphoglycerate (PGK) from Bacillus stearothermophilus NCA1503, has been cloned in Escherichia coli and its complete nucleotide sequence determined. The gene consists of an open reading frame corresponding to a protein of 394 amino acids (aa) (calculated Mr 42,703) and, in common with other prokaryotic pgk genes, is preceded by the structural gene encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Constructs containing the B. stearothermophilus pgk gene and its flanking sequences in the high-copy plasmid, pUC9, co-express both PGK and GAPDH at high levels in transformed E. coli cells, typically producing PGK at levels of up to 30% of the soluble cell protein. The deduced aa sequence of B. stearothermophilus PGK is compared with those of the mesophilic (yeast) and extreme thermophilic (Thermus thermophilus) enzymes since the crystal structure of these PGKs are known or are in the process of being determined. Changes in the sequences of the three enzymes, as they appear to relate to the enhancement of thermal stability, are discussed.

Amino Acid Sequence

A 23 kDa protein from rat sperm plasma membranes shows sequence similarity and phospholipid binding properties to a bovine brain cytosolic protein.

A 23 kDa protein that is a major component of rat epididymal secretions and sperm plasma membranes has been purified and partially sequenced. A data-base search revealed approximately 85% sequence similarity with a phosphatidyl-ethanolamine-binding protein from bovine brain cytosol. The rat 23 kDa protein also showed selective affinity for phosphatidylethanolamine (Kd = 1.6 x 10(-5) M) with lower activity towards phosphatidylinositol and phosphatidylcholine. It is suggested that differential affinity of protein antigens towards asymmetrically aligned phospholipids in sperm plasma membranes could account for their organisation into specific regional domains.

Amino Acid Sequence

cDNA cloning of porcine interleukin 2 by polymerase chain reaction.

Porcine interleukin 2 (IL-2) cDNA was cloned by polymerase chain reaction (PCR), using primers derived from the corresponding bovine sequence. The resulting porcine DNA sequence encodes a 154 residue IL-2 primary translation product. Comparison of the mature, secreted form of porcine IL-2 with those of other species was carried out in an attempt to identify differences that might contribute to the observed differing species specificities.

Amino Acid Sequence

Conformational changes in yeast phosphoglycerate kinase upon ligand binding: fluorescence of a linked probe and chemical reactivity of genetically introduced cysteinyl residues.

The effects of ligands on the conformation of yeast phosphoglycerate kinase were explored by introducing cysteinyl residues at different positions in the molecule by site-directed mutagenesis. Thus several mutants were constructed, each containing a unique cysteinyl residue. Neither the conformation nor the enzyme activity was affected by the substitutions. The reactivity of the thiol groups and the fluorescence of N-acetyl-N'-(5-sulfo-1-naphthyl)ethylene-diamine covalently linked to these thiols were used to monitor the conformational changes induced upon ligand binding. It was found that the observed changes mainly involve the part of the protein located in the cleft, particularly the environment of residues 35 and 183. No alteration was observed on the external side of the protein. Only 3-Phosphoglycerate induced these conformational changes. However, when the fluorescent probe was attached to residue 377, the binding of the two substrates was required to induce a modification in the fluorescence of the probe. These results indicate that the substrates separately or together induce discrete molecular motions in phosphoglycerate kinase.

Adenosine Triphosphate

Do children with severe head injury benefit from intensive care?

Intensive management of severe closed head injury is pursued in most neurosurgical units, as it no doubt reduces mortality. However, the fate of those salvaged from death is less clearly defined. The impact of aggressive management on the psychological recovery is even less clear. This retrospective study of children admitted to a regional paediatric neurosurgical service attempts to address these issues. Eighty-four children between the ages of 3 to 16 years with severe head injuries were studied and categorized according to conservative or intensive treatment groups. The availability of intensive management led to a greater number of such children referred. The mortality in those children sustaining more severe injury, i.e. Glasgow Coma Score (GCS) 3 or 4, was halved (44%) with intensive management from 80% in the conservative group. For those with less severe injury (GCS 5-8), it was unchanged at about 23%. The reduced mortality is at the expense of increased morbidity. Psychological testing showed that the cognitive recovery was comparable.

Adolescent

A genetic linkage map of human chromosome 5 with 60 RFLP loci.

A genetic map of human chromosome 5 that contains 60 restriction fragment length polymorphism (RFLP) loci in one linkage group has been constructed. Segregation data using these markers and 40 large multigenerational families supplied by the Centre d'Etude du Polymorphisme Humain have been collected. Linkage analyses were performed with the program package CRI-MAP; using odds greater than 1000:1, 30 RFLP loci could be placed on the map. This genetic map spans 289 cM sex-equal, 353 cM in females, and 244 cM in males. While the relative rate of recombination for female meioses is nearly twice that of males over much of the chromosome, several instances of statistically significant excess male recombination were observed. The order of probes on the genetic map has been confirmed by their physical order as determined by somatic cell hybrid lines containing deletions of normal chromosome 5. There is concordance between the physical positions of markers and their genetic positions. Our most distal probes on the genetic map are cytologically localized to the most distal portions of the chromosome. This suggests that our genetic map spans most of chromosome 5.

Blotting, Southern