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D P Brown

Publications and source records attributed to D P Brown.

At least 19 recordsLinked to original sources
PubMed1996

Insertional mutagenesis and recovery of interrupted genes of Streptococcus mutans by using transposon Tn917: preliminary characterization of mutants displaying acid sensitivity and nutritional requirements.

New vectors were constructed for efficient transposon Tn917-mediated mutagenesis of poorly transformable strains of Streptococcus mutans(pTV1-OK) and subsequent recovery of interrupted genes in Escherichia coli (pT21delta2TetM). In this report, we demonstrate the utility of Tn917 mutagenesis of a poorly transformable strain of S. mutans (JH1005) by showing (i) the conditional replication of pTV1-OK, a repA(Ts) derivative of the broad-host-range plasmid pWVO1 harboring Tn9l7, in JH1005 at the permissive temperature (30 degrees C) versus that at the nonpermissive temperature (45 degrees C); (ii) transposition frequencies similar to those reported for Bacillus subtilis (10(-5) to 10(-4)) with efficient plasmid curing in 90 to 97% of the erythromycin-resistant survivors following a temperature shift to 42 to 45 degrees C; and (iii) the apparent randomness of Tn917 insertion as determined by Southern hybridization analysis and the ability to isolate nutritional mutants, mutants in acid tolerance, and mutants in bacteriocin production, at frequencies ranging from 0.1 to 0.7%. Recovery of transposon-interrupted genes was achieved by two methods: (i) marker rescue in E. coli with the recovery vector pTV21delta2TetM, a tetracycline-resistant and ampicillin-sensitive Tn9l7-pBR322 hybrid, and (ii) "shotgun" cloning of genomic libraries of Tn917 mutants into pUC19. Sequence analyses revealed insertions at five different genetic loci in sequences displaying homologies to Clostridium spp.fhs (66% identity), E. coli dfp (43% identity), and B. subtilis ylxM-ffh (58% identity), icd (citC [69% identity]), and argD (61% identity). Insertions in icd and argD caused nutritional requirements; the one in ylxM-ffh caused acid sensitivity, while those in fhs and dfp caused both acid sensitivity and nutritional requirements. This paper describes the construction of pTV1-OK and demonstrates that it can be efficiently employed to deliver Tn917 into S. mutans for genetic analyses with some degree of randomness and that insertions in the chromosome can be easily recovered for subsequent characterization. This represents the first published report of successful Tn9l7 mutagenesis in the genus Streptococcus.

Bacterial Proteins↗
PubMed1994

Characterization of the macromolecular synthesis (MMS) operon from Listeria monocytogenes.

The macromolecular synthesis (MMS) operon consists of three genes: rpsU, which encodes the S21 ribosomal protein in Bacillus subtilis (Bs), rpsU is replaced by orfP23 which encodes a protein of unknown function), dnaG, encoding the DNA primase involved in the initiation of chromosome replication, and rpoD, which encodes the principal sigma subunit of RNA polymerase. The operon was cloned in three segments from Listeria monocytogenes (Lm), initially using a probe designed from a highly conserved region of RpoD. Analysis of the nucleotide sequence revealed three genes: orfP17 (whose product, P17, is homologous to Bs P23), dnaG and rpoD. The Lm DnaG resembles the primase from Escherichia coli through the first two-thirds of the sequence. C-terminal similarity was observed between DnaG from Lm and Bs. Lm RpoD is similar to Bs SigA, shares identical DNA-binding domains with SigA, and is a member of the sigma 43 subgroup of the sigma 70 family.

Amino Acid Sequence↗
PubMed1994

Follow-up study of chrysotile asbestos textile workers: cohort mortality and case-control analyses.

Previous studies of mortality among white males employed in a Charleston, South Carolina asbestos textile plant using chrysotile demonstrated significant excess mortality due to asbestos-related disease and a steep exposure-response relationship for lung cancer. This cohort was further studied by adding 15 years of follow-up and including mortality among white female and black male workers. Nested case-control analyses were undertaken to further explore possible differences in lung cancer risk by textile operation as well as possible confounding by mineral oil exposures. Preliminary data for white males have been previously published. White males experienced statistically significant excess mortality due to lung cancer (standardized mortality ratio [SMR] = 2.30; confidence interval [CI] = 1.88-2.79), all causes (SMR = 1.48; CI = 1.38-158), all cancers (SMR = 1.50; CI = 1.29-1.72), diabetes mellitus (SMR = 2.05; CI = 1.18-3.33), heart disease (SMR = 1.41; CI = 1.26-1.58), cerebrovascular disease (SMR = 1.50; CI = 1.08-2.02), pneumoconiosis and other respiratory diseases (SMR = 4.10; CI = 3.10-5.31), and accidents (SMR = 1.49; CI = 1.15-1.91). Among white females, statistically significant excesses occurred for lung cancer (SMR = 2.75; CI = 2.06-3.61), all causes (SMR = 1.21; CI = 1.11-1.32), pneumoconiosis and other respiratory diseases (SMR = 2.40; CI = 1.53-3.60), and other respiratory cancers (SMR = 14.98; CI = 4.08-38.7). Among the total cohort of black males, the only statistically significant excess observed was for pneumoconiosis (SMR = 2.19; CI = 1.23-3.62). Based on historical exposure measurements at the plant, there was a positive exposure-response relationship for both lung cancer and pneumoconiosis. Data for the entire cohort demonstrate an increase in the lung cancer relative risk of 2-3% for each fiber/cc-year of cumulative chrysotile exposure. This relationship was more consistent for the white male workers. The excess risk for lung cancer among white males and females appeared to occur at cumulative exposures lower than those for black males. Possible reasons for the lesser lung cancer risk among black males include less smoking and differences in airborne fiber characteristics experienced by black males as a result of plant job placement patterns. The case-control analysis found employment in preparation and carding operations (where most of the black males worked) to be associated with a slightly reduced lung cancer risk, although not statistically significant, whereas spinning and twisting employment was associated with a statistically significant increased lung cancer risk compared to other plant operations.(ABSTRACT TRUNCATED AT 400 WORDS)

Black or African American↗
PubMed1994

Characterization of the genes and attachment sites for site-specific integration of plasmid pSE101 in Saccharopolyspora erythraea and Streptomyces lividans.

The 11.3 kb plasmid pSE101 integrates into the chromosome of Saccharopolyspora erythraea at a specific attB site and into the chromosome of Streptomyces lividans at many sites. Multisite integration in S. lividans was also observed when a 1.9 kb segment of pSE101 containing attP and adjacent plasmid sequence was used to transform a pSE101- S. lividans host. Nucleotide sequencing of this segment revealed the presence of a complete open reading frame (ORF) designated int, encoding a putative polypeptide of 448 amino acids that shows similarities to site-specific recombinases of the integrase family. Sequencing of the 1.3 kb segment upstream of int revealed the presence of three additional ORFs: the one most distal to int encodes a putative 76 amino acid basic polypeptide analogous to the Xis proteins of a number of bacteriophages. Nucleotide sequencing of attP, and the attB, attL and attR sites from Sac. erythraea revealed a 46 bp sequence common to all sites with no duplications of chromosomal sequences in the integrated state. A putative structural gene for a tRNA(Thr) was found to overlap the 46 bp common sequence at attB. Sequencing of four pSE101 integration sites (attB') and corresponding attL' and attR' sites in S. lividans showed that the 46 bp sequence was present at each attR' site, whereas only the first three bases, CTT, were retained at each attL' and attB' site. A feature common to the four attB' sites and to attB is a highly conserved 21 bp segment with inverted repeats flanking the CTT sequence.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1994

Lung cancer mortality among asbestos textile workers: a review and update.

In an update of the mortality of the cohort of 1200 South Carolina textile workers, of whom almost half died, there were 185 excess deaths (SMR = 1.44), which included 71 cardiovascular diseases (SMR = 1.37), 43 non-malignant respiratory diseases (SMR = 2.25) and 41 lung cancers (SMR = 2.25). Only two definite mesotheliomas were observed. Other possible cases may have occurred but no confirmatory pathology was available. Strong exposure-response relationships have been found for lung cancer and for non-malignant respiratory diseases. The data suggest a doubling of the lung cancer risk at an exposure of approximately 30 fibre years. Mortality from pneumoconiosis and other respiratory diseases was elevated at even the lowest cumulative exposure category (< 2 f ml-1 years). A nested case-control analysis failed to demonstrate a significant role for mineral oil exposure in the etiology of lung cancer. Differences in airborne fibre sizes may be important in explaining different lung cancer and pneumoconiosis risks in various industries. In particular, the data on airborne fibres in textile manufacturing industries suggested 11-27% were longer than 5 microns compared to 2-5% for mining and milling.

Asbestos, Serpentine↗
PubMed1994

Characterization of spo0A homologues in diverse Bacillus and Clostridium species identifies a probable DNA-binding domain.

Spo0A is a phosphorylation-activated transcription factor of Bacillus subtilis. It is a member of the response regulator superfamily of bacterial signal transduction proteins and controls many of the changes in gene expression that occur during the transition into stationary phase and during the initiation of sporulation. To identify the domains of Spo0A most critical for determining its structural and functional features, presumptive homologues of the spo0A gene were characterized in a collection of eight Bacillus species and six Clostridium species representing phylogenetically diverse members of these genera. An alignment of the partial or complete DNA sequences of these homologues revealed three regions of especially high conservation in the effector domain. We speculate that the most highly conserved of these corresponds to the recognition helix of a putative helix-turn-helix motif, and, therefore, represents the actual DNA-containing surface of the protein. In the case of homologues identified in Bacillus anthracis and Clostridium acetobutylicum and retrieved by polymerase chain reaction amplification, we confirmed by gene-disruption analysis that the homologue actually is required for initiation of sporulation. Apparent homologues of the B. subtilis spoIVB gene were also discovered immediately upstream from the spo0A homologues in all Bacillus and Clostridium species examined. The discovery of homologues of B. subtilis sporulation genes in these diverse species implies that the gene products required for specifying pathways of sporulation-specific gene activation and for determining key morphogenetic changes may be highly conserved and suggests that an approach similar to that undertaken here might be used as a general strategy to retrieve and compare their gene sequences. Exhaustive efforts to detect a spo0A-like gene in non-endospore formers, including close relatives of Bacillus such as Listeria and Staphylococcus, were uniformly unsuccessful, suggesting that regulation of gene activity during the transition into stationary phase mediated by Spo0A-like proteins may be exclusive to the endospore-forming bacteria.

Amino Acid Sequence↗
PubMed1994

Auditory P300 abnormalities and leukocyte activation in HIV infection.

To evaluate whether P300 testing might serve as a screening modality for the early detection of HIV-related neuropathology, we tested 26 HIV-infected men (23 without neurologic symptoms, 2 with peripheral neuropathy, 1 with AIDS-associated dementia) and 15 controls. Although they had no overt neurologic symptoms, the P300 latency was delayed or undetectable in 30% of patients without clinically evident neurologic disease. P300 latencies did not correlate with peripheral blood CD4 T-cell count, serum quinolinic acid or p24 antigen levels, or the numbers of activated peripheral blood monocytes. Three individuals with abnormal P300 latencies had been HIV-seropositive for < or = 1 year, suggesting that delayed evoked responses detect early neurologic dysfunction. P300 responses do not predict imminent dementia. In only one previously asymptomatic individual with abnormal P300 waveforms have overt neurologic symptoms developed during a 2-year followup. Extended longitudinal studies will be necessary to define the predictive value of P300 latencies in the development of AIDS-related dementia. However, the sensitivity, quantitative nature, and speed of administration of this test suggest that it may be useful for identification of early neurologic involvement in HIV infection.

Acoustic Stimulation↗
PubMed1994

Cancer mortality in female and male dry-cleaning workers.

A cohort study of dry-cleaning workers (1109 women, 592 men) in the mid-1980s revealed significant excess bladder cancer mortality. This article updates vital status through 1990. Significant excesses were seen for bladder cancer (nine deaths, standardized mortality ratio [SMR] = 2.54, 95% confidence interval [CI] = 1.16-4.82), esophageal cancer (10 deaths, SMR = 2.14, 95% CI = 1.02-3.94), and intestinal cancer (26 deaths, SMR = 1.56, 95% CI = 1.02-2.29). In a subcohort exposed only to perchloroethylene (PCE), those with 5 or more years of employment and 20 or more years since first exposure had a significant increased risk of esophageal cancer (four deaths, SMR = 7.17, 95% CI = 1.92-19.82). Women had significant excess esophageal cancer (five deaths, SMR = 3.24, 95% CI = 1.05-7.58) and elevated SMRs for intestinal, pancreatic, and bladder cancer mortality. This study confirms the esophageal cancer risk among dry-cleaning workers seen in another study and suggests an association with PCE. It further documents the risks for intestinal, pancreatic, and bladder cancers in this industry.

Cause of Death↗
PubMed1994

Mortality patterns among female and male chrysotile asbestos textile workers.

This study updates a retrospective cohort mortality analysis of workers from a South Carolina textile plant where chrysotile asbestos was the primary exposure. The update adds 15 years of observation to the original study, adds analyses of white women and black men, and allows comparison of mortality risks between race/gender groups. The total cohort includes 3,022 workers: 1,229 white women (363 deaths), 1,247 white men (607 deaths), and 546 black men (289 deaths). Statistically significant risks for lung cancer were observed among white women (standardized mortality ratio [SMR] = 2.07; 90% confidence interval [CI] = 1.55-2.71) and white men (SMR = 2.24; 90% CI = 1.83-2.72); both of these groups exhibited positive exposure-response trends. Although the lung cancer risk among black men was lower than expected (SMR = 0.70; 90% CI = 0.42-1.08), a statistically significant increase was observed at high levels of exposure. Statistically significant excess risk for pneumoconiosis and other respiratory diseases were observed for all race/gender groups. Despite the relatively high percentage of white women lost to follow-up and missing death certificates, both of which allow underestimation of the true relative risk, statistically significant excess risks were observed for lung cancer and pneumoconiosis among this group.

Asbestos, Serpentine↗
PubMed1994

Speech recognition in recurrent otitis media: results in a set of identical twins.

Performance-intensity functions for Pediatric Speech Intelligibility Test (PSI) word and sentence materials presented in competition were obtained for a set of identical female twins, aged 4 years, 2 months. Twin B had a medical history significant for recurrent otitis media, while Twin A had only one reported occurrence of the disease. Twin A yielded normal functions for both word and sentence materials. Twin B, however, yielded an essentially normal function for sentences, but the function for monosyllabic words was grossly depressed. These findings support the hypothesis that transient conductive hearing loss and auditory deprivation resulting from recurrent otitis media can affect the processing of phonetic information essential to the development of word recognition skills.

Audiometry, Pure-Tone↗
PubMed1992

High-level expression and purification of mature HIV-1 protease in Escherichia coli under control of the araBAD promoter.

A 1.3-kb segment of Escherichia coli DNA containing the regulatory gene, araC, and the promoter of the araBAD operon was amplified by the polymerase chain reaction (PCR) and cloned into pUC18, resulting in plasmid pKB130 that produced the alpha fragment of beta-galactosidase upon addition of L-arabinose (L-ara). A synthetic gene for human immunodeficiency virus (HIV)-1 preprotease was placed downstream of the ara-BAD promoter in pKB130 to create a translational fusion inducible by addition of L-ara. The fusion protein correctly autoprocessed in vivo to yield a mature 99-amino-acid HIV-1 protease, which was found predominantly in inclusion bodies. This material could be refolded to an active form, which was purified to homogeneity. A small fraction of the protease was expressed in vivo as a soluble active form, which allowed the monitoring of expression during fermentation by a rapid and simple whole cell assay employing an HIV-1 protease-specific fluorogenic substrate.

Amino Acid Sequence↗
PubMed1992

Mortality of workers employed at organochlorine pesticide manufacturing plants--an update.

A previous mortality study among four organochlorine pesticide manufacturers was updated through 1987. The organochlorine pesticides included chlordane; heptachlor and endrin; aldrin, dieldrin and endrin; and dichlorodiphenyltrichloroethane. The mortality for all causes and all malignant neoplasms at each of the plants was lower than expected. There was a statistically significant increase in liver and biliary tract cancer among workers at plant 3 (5 observed, standardized mortality ratio 393, 95% confidence interval 1.27-9.20). These results are somewhat consistent with experimental animal findings showing benign and malignant tumors of the liver after exposure to aldrin and dieldrin. However, the deaths were due to a mixture of intra- and extra-hepatic tumors, and the dose-response analysis was limited because of the small number of deaths and lack of exposure data. Additional study of this group should include continued follow-up of the total cohort and a case-referent analysis of the deaths from liver and biliary tract cancer.

Bile Duct Neoplasms↗
PubMed1991

Site-specific recombination in Escherichia coli between the att sites of plasmid pSE211 from Saccharopolyspora erythraea.

pSE211 from Saccharopolyspora erythraea integrates site-specifically into the chromosome through conservative recombination between attP and attB, the plasmid and chromosomal attachment sites. Integration depends on the presence of int, an open reading frame (ORF) that lies adjacent to attP and encodes the putative integrase. Immediately upstream of int lies xis (formerly called orf2) which encodes a basic protein that is thought to exhibit DNA binding. xis and int were cloned in various combinations in pUC18 and expressed constitutively in Escherichia coli from the lac promoter. attP and attB were cloned in Streptomyces or E. coli plasmids containing kanamycin resistance (KmR) or chloramphenicol resistance (CmR) markers. Stable KmR CmR cointegrates formed by attP x attB or attP x attP recombination (integration) were obtained in E. coli hosts that expressed int. Co-integrates were not found in hosts expressing int + xis. Excision (intraplasmid att site recombination) was examined by constructing plasmids carrying attL and attR or two attP sites separating CmR from KmR and by following segregation of the markers in various hosts. Both attL x attR and attP x attP excision depended on both xis and int in E. coli. pSE211 att site integration and excision were not affected by a deletion in himA, the gene encoding a subunit of integration host factor.

Actinomycetales↗
PubMed1991

Audiologic findings in a set of fraternal twins with CHARGE association.

CHARGE association refers to a constellation of congenital malformations of unknown etiology that includes ocular Coloboma, Heart defects, choanal Atresia, Retarded growth/development, Genital hypoplasia, and Ear anomalies and/or hearing loss. This paper discusses audiologic results in a set of fraternal twins with a diagnosis of CHARGE. Both infants exhibit external ear defects and audiologic patterns characteristic of CHARGE and demonstrate the variable and often asymmetric findings associated with this disorder. Early identification and evaluation are imperative for appropriate audiologic management of individuals with a diagnosis of CHARGE and for persons with CHARGE-like anomalies.

Abnormalities, Multiple↗
PubMed1990

Evidence for an effect of ELF electromagnetic fields on human pineal gland function.

A study was carried out to determine possible effects of 60-Hz electromagnetic-field exposure on pineal gland function in humans. Overnight excretion of urinary 6-hydroxymelatonin sulfate (6-OHMS), a stable urinary metabolite of the pineal hormone melatonin, was used to assess pineal gland function in 42 volunteers who used standard (conventional) or modified continuous polymer wire (CPW) electric blankets for approximately 8 weeks. Volunteers using conventional electric blankets showed no variations in 6-OHMS excretion as either a group or individuals during the study period. Serving as their own controls, 7 of 28 volunteers using the CPW blankets showed statistically significant changes in their mean nighttime 6-OHMS excretion. The CPW blankets switched on and off approximately twice as often when in service and produced magnetic fields that were 50% stronger than those from the conventional electric blankets. On the basis of these findings, we hypothesize that periodic exposure to pulsed DC or extremely low frequency electric or magnetic fields of sufficient intensity and duration can affect pineal gland function in certain individuals.

Adult↗
PubMed1990

Characterization of the genetic elements required for site-specific integration of plasmid pSE211 in Saccharopolyspora erythraea.

The 18.1-kilobase plasmid pSE211 integrates into the chromosome of Saccharopolyspora erythraea at a specific attB site. Restriction analysis of the integrated plasmid, pSE211int, and adjacent chromosomal sequences allowed identification of attP, the plasmid attachment site. Nucleotide sequencing of attP, attB, attL, and attR revealed a 57-base-pair sequence common to all sites with no duplications of adjacent plasmid or chromosomal sequences in the integrated state, indicating that integration takes place through conservative, reciprocal strand exchange. An analysis of the sequences indicated the presence of a putative gene for Phe-tRNA at attB which is preserved at attL after integration has occurred. A comparison of the attB site for a number of actinomycete plasmids is presented. Integration at attB was also observed when a 2.4-kilobase segment of pSE211 containing attP and the adjacent plasmid sequence was used to transform a pSE211- host. Nucleotide sequencing of this segment revealed the presence of two complete open reading frames (ORFs) and a segment of a third ORF. The ORF adjacent to attP encodes a putative polypeptide 437 amino acids in length that shows similarity, at its C-terminal domain, to sequences of site-specific recombinases of the integrase family. The adjacent ORF encodes a putative 98-amino-acid basic polypeptide that contains a helix-turn-helix motif at its N terminus which corresponds to domains in the Xis proteins of a number of bacteriophages. A proposal for the function of this polypeptide is presented. The deduced amino acid sequence of the third ORF did not reveal similarities to polypeptide sequences in the current data banks.

Amino Acid Sequence↗
PubMed1990

Environmental epidemiologic investigations in the styrene-butadiene rubber production industry.

A review of the literature and an update that is in progress of a previous retrospective cohort mortality study of the styrene-1,3-butadiene industry are discussed. The follow-up has now been extended from April 1, 1976, through December 31, 1981, for plant B and December 31, 1982, for plant A. The person-years at risk of death have gone from 34,187 to 43,341 in plant A and from 19,742 to 26,314 in plant B. Among the death certificates received to date, observed deaths have increased in both plants, with increases in cancers of the trachea, bronchus and lung and in lymphosarcomas, reticulosarcomas, and cancers of the overall lymphatic and hematopoietic system.

Bronchial Neoplasms↗