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C W Bell

Publications and source records attributed to C W Bell.

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Characterization of the glycoprotein D gene products of equine herpesvirus 1 using a prokaryotic cell expression vector.

The gene encoding equine herpesvirus 1 (equine abortion virus; EHV-1) glycoprotein D was engineered into the prokaryotic vector pEX, and expressed as a beta-galactosidase fusion product, which was recognized by pooled equine sera and anti-EHV-1 rabbit sera. Antibodies raised against the EHV-1 gD fusion product identified strong bands in infected cells at 66 and 68 K and at 138 K in purified virus, thus characterizing the several forms of this major envelope glycoprotein which is an important candidate for inclusion in subunit vaccines.

Animals

Sequence characteristics of a gene in equine herpesvirus 1 homologous to glycoprotein H of herpes simplex virus.

A gene in equine herpesvirus 1 (EHV-1, equine abortion virus) homologous to the glycoprotein H gene of herpes simplex virus (HSV) was identified and characterised by its nucleotide and derived amino acid sequence. The EHV-1 gH gene is located at 0.47-0.49 map units and contains an open reading frame capable of specifying a polypeptide of 848 amino acids, including N- and C-terminal hydrophobic domains consistent with signal and membrane anchor regions respectively, and 11 potential sites for N-glycosylation. Alignment of the amino acid sequence with those published for HSV gH, varicella zoster virus gpIII, Epstein Barr virus gp85 and human cytomegalovirus p86 shows similarity of the EHV gene with the 2 other alpha-herpesviruses over most of the polypeptide, but only the C-terminal half could be aligned for all 5 viruses. The identical positioning of 6 cysteine residues and a number of highly conserved amino acid motifs supports a common evolutionary origin of this gene and is consistent with its role as an essential glycoprotein of the herpesvirus family. An origin of replication is predicted to occur at approximately 300 nucleotides downstream of the EHV-1 gH coding region, on the basis of similarity to other herpesvirus origins.

Amino Acid Sequence

Transcript analysis of the equine herpesvirus 1 glycoprotein B gene homologue and its expression by a recombinant vaccinia virus.

Transcript mapping of the equine herpesvirus 1 (EHV-1) glycoprotein B (gB) gene homologue by Northern blot, S1 nuclease and primer extension analyses indicated that two overlapping transcripts of 3.4 and 4.6 kb originated from the same strand and were transcribed from left to right between coordinates 0.40 and 0.43 of the EHV-1 genome. The 3.4 kb transcript encoded EHV-1 gB and the 5' RNA terminus was located approximately 30 bases downstream from a probable TATA element. The coding region of the gB gene homologue was reconstructed from two subclones using oligonucleotide mutagenesis and inserted into vaccinia virus by homologous recombination. Cells infected with the recombinant virus synthesized EHV-1 gB antigen, which was detectable in the cytoplasm and on the cell surface by immunofluorescence using an EHV-1 neutralizing horse serum and EHV-1 monoclonal antibodies. On Western blots, bands of 138K to 143K, 80K to 90K and 55K to 57K were identified in recombinant virus-infected cells, by both EHV-1 monoclonal antibodies and the polyclonal horse serum. These were similar in Mr to bands identified by these sera in EHV-1-infected cells. Mice vaccinated with the recombinant virus produced antibodies which recognized proteins of the same Mr as EHV-1 gB, on Western blots, but did not have in vitro neutralizing activity.

Animals

Smell and taste function in subjects with chronic obstructive pulmonary disease. Effect of long-term oxygen via nasal cannulas.

It was hypothesized that subjects with chronic obstructive pulmonary disease (COPD) receiving long-term oxygen via nasal cannulas have an impaired sense of smell and/or taste. To objectively evaluate the sense of smell and taste, this study used the University of Pennsylvania Identification Test (UPSIT), a 40-item "scratch-n-sniff" test and a 20-item taste test using the four basic taste sensations of sweet, salt, sour, and bitter. Twenty subjects (15 male, 5 female) with severe COPD receiving long-term oxygen therapy (group 1), and an equal number of age- and sex-matched subjects with COPD not receiving oxygen therapy (group 2), and a healthy control group (group 3) were studied. Twelve subjects (seven male, five female) from group 1 subsequently underwent transtracheal oxygen catheter installation. Mean +/- SD for the basic smell test was significantly greater in group 3 (35.35 +/- 3.58) as compared with group 1 (27.70 +/- 6.07) or group 2 (31.10 +/- 4.95) (p less than 0.005). The difference between group 1 and 2 was not significant (p = 0.066). However, when adjusted for pack years of smoking, there were no significant differences between the three groups. Mean +/- SD correct responses for the basic taste test were significantly greater in group 3 (15.75 +/- 1.81) as compared with group 2 (12.8 +/- 2.78) (p less than 0.005) and group 1 (14.00 +/- 2.33) (p less than 0.05). There was no significant difference between group 1 and 2. The corrected data for taste, adjusted for years since quitting smoking, did not alter the basic differences between the groups. Mean smell and taste test scores were essentially unchanged in 12 subjects after six months of transtracheal oxygen therapy. Long-term oxygen use via nasal cannulas in this group of subjects with COPD did not appear to impair their sense of smell and taste. Smoking had a significant but variable effect on the sense of smell and taste.

Female

Analysis of the length of S-phase required to show sister chromatid differential staining.

In vitro studies of BrdU-dependent sister chromatid differential staining typically employ two cycles of BrdU incorporation. Experiments are described which determined the actual fraction of both S-phases that the rat embryonic fibroblasts (Rat-1) cells had to traverse in order to show distinctive differential staining. Following synchronization of cells by a combination of serum deprivation and hydroxyurea blockage, sister chromatid differential staining, labelling index, mitotic index, and per cent DNA replication are determined. Results indicate that only approximately 50% of the first S-phase is necessary in order to show distinctive differential staining. The importance of this finding to studies of cellular proliferation using BrdU incorporation is discussed.

Animals

Polypeptide subunits of dynein 1 from sea urchin sperm flagella.

A high-resolution sodium dodecyl sulfate polyacrylamide gel electrophoresis system has been used to show the presence, in both whole sperm and isolated flagellar axonemes, of eight polypeptides migrating in the 300,000--350,000 molecular weight range characteristic of the heavy chains of dynein ATPase. Previously, only five such chains have been discernible. Extraction of isolated axonemes for 10 min at 4 degrees C with a solution containing 0.6 M NaCl, ph 7, releases a mixture of particles that separate, in sucrose density gradient centrifugation, into a major peak, dynein 1 ATPase, sedimenting at 21S and a minor peak at 12--14S. The polypeptide compositions of these two peaks are different. The dynein 1 peak, which contains most of the protein on the gradient, contains approximately equal quantities of two closely migrating heavy chains, with a small amount of a third, more slowly migrating chain; no other heavy chains appear in this peak. Two groups of smaller polypeptides (three intermediate chains, within the apparent molecular weight range 76,000--122,000 and four newly discovered light chains, within the apparent molecular weight range 14,000--24,000) cosediment with the 21S peak. The heavy chain composition of the 12--14S peak is more complex, all eight heavy chains occurring approximately the same ratios as occur in intact axonemes.

Adenosine Triphosphatases

Immediate carbon monoxide estimates and self-reported smoking.

Because carbon monoxide (CO) is an easily understood danger of cigarette smoking, new breath CO analyzers may be useful in cessation programs. Self-reported cigarette consumption and breath CO correlated .70 using a 90-sec. CO-determination method. Smokers averaging 17 daily cigarettes had average breath CO of 24 parts per million (ppm), falling to 7.71 ppm for smokers claiming at least 24-hr. abstinence. Corresponding carboxyhemoglobin (COHb) estimates were 4.34% (slightly above World Health Organization 4% safety limit) and 1.39%. Smoking clinics can use CO-monitoring devices for feedback or surveillance, although feedback would provide the most benefits for participants.

Adolescent

Neurofilaments.

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Animals