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Biomedical subjects

W C Black

Publications and source records attributed to W C Black.

At least 19 recordsLinked to original sources

Phylogenetic relationships among tick subfamilies (Ixodida: Ixodidae: Argasidae) based on the 18S nuclear rDNA gene.

Phylogenetic relationships among tick subfamilies have been estimated using morphological and molecular characters. However, the phylogeny based on a portion of the 16S mitochondrial rDNA gene differed from the morphologically based phylogeny in a number of important respects. The entire 18S rDNA gene was examined in 18 taxa from all tick subfamilies to test the 16S rDNA based phylogeny. The 18S phylogeny supports the earlier 16S based phylogeny in placing members of Hyalomminae on a common branch with members of the Rhipicephalinae and in indicating long branch lengths among soft tick taxa. However, unlike the 16S phylogeny, Amblyomminae was monophyletic and members of Haemaphysalinae did not arise within Amblyomminae. Argasinae formed a monophyletic group within Argasidae and was not a sister taxon of the hard ticks. In most respects, the phylogeny based on the 18S rDNA gene resembles the morphologically based phylogeny.

Animals

Screening for disease.

Screening for disease has become a major activity over the last few decades, and the potential for future growth is almost boundless. However, the necessity for and effectiveness of screening has often been overstated with misused survival statistics. Nevertheless, screening with certain radiologic tests may provide modest benefits at acceptable costs under certain conditions. The challenge for the future is to better identify the tests, populations, and other conditions under which screening is appropriate. Various quantitative techniques for eliciting patient preferences [76] and analyzing benefits, harms, and costs over time [77-80] may help us meet this challenge.

Diagnostic Imaging

Pleural fluid volume estimation: a chest radiograph prediction rule.

RATIONALE AND OBJECTIVES: We devised a prediction rule for estimating pleural effusion volume on the basis of posteroanterior and lateral chest radiographs. METHODS: A prediction rule was devised for estimating pleural effusion volume on the basis of the presence or absence of a meniscus on chest radiographs. The rule was tested and validated using separate data sets obtained from a retrospective review of patients having both a chest radiograph and computed tomography (CT) scan (the gold standard) within 24 hr of each other. The accuracy of the prediction rule and the degree of interobserver agreement between the two independent readers were determined. RESULTS: For the test and validation sets, the weighted accuracies of the prediction rule were 86% and 85%, respectively. The respective weighted interobserver agreements were 97% and 88%. Pleural effusions became visible as a meniscus on the lateral chest radiograph at a volume of approximately 50 ml; at a volume of 200 ml, the meniscus could be identified on the posteroanterior radiograph. At a volume of about 500 ml, the meniscus obscured the hemidiaphragm. CONCLUSION: The volume of a pleural effusion can be estimated from the chest radiograph appearance with a reasonable degree of accuracy.

Humans

Population genetics with RAPD-PCR markers: the breeding structure of Aedes aegypti in Puerto Rico.

RAPD-PCR polymorphisms at 57 presumptive loci were used to examine the breeding structure of the mosquito Aedes aegypti in Puerto Rico. Mosquitoes were sampled from 16 locations in six cities and samples were located in a nested spatial design to examine local patterns of gene flow. Allele frequencies were estimated assuming (1) that genomic regions amplified by RAPD-PCR segregate as dominant alleles, (2) that genotypes at RAPD loci are in Hardy-Weinberg proportions, (3) identity in state (iis) among dominant amplified alleles and (4) iis among null alleles. The average genic heterozygosity was 0.354, more than twice the level detected in earlier allozyme surveys. Nested analysis of variance indicated extensive genetic differentiation among locations within cities. Effective migration rates (Nm) among cities were estimated from FST assuming an island model of migration. Estimates of Nm ranged from 9.7 to 12.2 indicating a high dispersal rate. The large number of polymorphisms revealed by RAPD-PCR allowed the distribution of FST and linkage disequilibrium to be examined among loci and demonstrated that small samples inflate FST and linkage disequilibrium. No linkage disequilibrium maintained through epistasis was detected among alleles at the 57 loci.

Aedes

Intensive linkage mapping in a wasp (Bracon hebetor) and a mosquito (Aedes aegypti) with single-strand conformation polymorphism analysis of random amplified polymorphic DNA markers.

The use of random amplified polymorphic DNA from the polymerase chain reaction (RAPD-PCR) allows efficient construction of saturated linkage maps. However, when analyzed by agarose gel electrophoresis, most RAPD-PCR markers segregate as dominant alleles, reducing the amount of linkage information obtained. We describe the use of single strand conformation polymorphism (SSCP) analysis of RAPD markers to generate linkage maps in a haplodiploid parasitic wasp Bracon (Habrobracon) hebetor and a diploid mosquito. Aedes aegypti. RAPD-SSCP analysis revealed segregation of codominant alleles at markers that appeared to segregate as dominant (band presence/band absence) markers or appeared invariant on agarose gels. Our SSCP protocol uses silver staining to detect DNA fractionated on large thin polyacrylamide gels and reveals more polymorphic markers than agarose gel electrophoresis. In B. hebetor, 79 markers were mapped with 12 RAPD primers in six weeks; in A aygpti, 94 markers were mapped with 10 RAPD primers in five weeks. Forty-five percent of markers segregated as codominant loci in B. hebetor, while 11% segregated as codominant loci in A. aegypti. SSCP analysis of RAPD-PCR markers offers a rapid and inexpensive means of constructing intensive linkage maps of many species.

Aedes

Population genetics of Ixodes scapularis (Acari: Ixodidae) based on mitochondrial 16S and 12S genes.

Blacklegged tick, Ixodes scapularis Say, is the principal vector of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner in the eastern half of the United States. Populations exhibit extreme variation in morphology, host usage, development time, and behavior. We examined sequence variation in the 16S and 12S mitochondrial ribosomal DNA genes to determine genetic relationships among I. scapularis collections from throughout its range. Single strand conformation polymorphism analysis of 300 bp of the 16S molecule was used to identify different haplotypes and estimate their relative frequencies among 198 ticks. Eleven different haplotypes were detected. Haplotype diversity was least in northeastern collections and greatest in the southeast. The 11 haplotypes were sequenced in 24 specimens. In total, 462 bp in the 16S gene and 420 bp in the 12S gene were sequenced to reveal 66 informative sites. Phylogenetic analysis, using I. ricinus L. and I. pacificus Cooley & Kohls as outgroups, revealed 2 clades within I. scapularis. One clade was limited to the South and the other was distributed throughout the range of I. scapularis. Specimens from the Southern United States were basal in the broadly distributed clade. Random amplified polymorphic DNA by polymerase chain reaction patterns examined between members of the 2 clades provided no evidence for reproductive isolation. These patterns suggest that I. scapularis arose in the South but that a large geographic split gave rise to 2 distinct lineages. These lineages now interbreed and are partially sympatric.

Animals

Evolution of ticks.

Evolutionary patterns in ticks have traditionally been cast in terms of host associations. Largely untested assumptions of cospeciation and observations of current host associations are used to estimate the age of different taxa. Several recent phylogenetic studies of supraspecific relationships in ticks, based on both morphological and DNA-sequence data, allow the first rigorous testing of these assumptions. Reanalysis of patterns of tick-host associations suggests that the perception of host specificity in ticks may be an artifact of incomplete sampling. An analysis of tick-host and -habitat associations and biogeographical patterns, in the context of the newly derived phylogenies, suggests that much of the existing host-association patterns may be explained as artifacts of biogeography and ecological specificity.

Animals

Cyclooxygenase-2 inhibitors. Synthesis and pharmacological activities of 5-methanesulfonamido-1-indanone derivatives.

The recent discovery of an alternative form cyclooxygenase (cyclooxygenase-2, COX-2), which has been proposed to play a significant role in inflammatory conditions, may provide an opportunity to develop anti-inflammatory drugs with fewer side effects than existing non-steroidal anti-inflammatory drugs (NSAIDs). We have now identified 6-[(2,4-difluorophenyl)-thio]-5-methanesulfonamido-1-indanone++ + (20) (L-745,337) as a potent, selective, and orally active COX-2 inhibitor. The structure-activity relationships in this series have been extensively studied. Ortho- and para-substituted 6-phenyl substitutents are optimal for in vitro potency. Replacement of this phenyl ring by a variety of heterocycles gave compounds that were less active. The methanesulfonamido group seems to be the optimal group at the 5-position of the indanone system. Compound 20 has an efficacy profile that is superior or comparable to that of the nonselective COX inhibitor indomethacin in animal models of inflammation, pain, and fever and appears to be nonulcerogenic within the dosage ranges required for functional efficacy. Although 20 and its oxygen linkage analog 2 (flosulide) are equipotent in the in vitro assays, compound 20 is more potent in the rat paw edema assay, has a longer t1/2 in squirrel monkeys, and seems less ulcergenic than 2 in rats.

Animals

Perceptions of breast cancer risk and screening effectiveness in women younger than 50 years of age.

BACKGROUND: The National Cancer Institute has recently changed its approach and has substituted summary-of-evidence statements for specific recommendations for breast cancer screening in women 40-50 years of age, leaving these women with a greater share of decision-making responsibility. To make an informed decision about breast cancer screening, women need accurate information about their breast cancer risk and the benefit of screening. Although it has been suggested that women younger than 50 years of age overestimate this risk and benefit, their estimates have not been quantified. PURPOSE: The purpose of this study was to determine how women 40-50 years of age perceive their risk of breast cancer and the effectiveness of screening and how these perceptions compare with estimates derived from epidemiologic studies of breast cancer incidence and randomized clinical trials of screening. METHODS: We mailed a questionnaire to 200 women, identified through the computerized medical records of Dartmouth-Hitchock Medical Center, who were between 40 and 50 years of age and had no history of breast cancer. Each woman was asked about her risk factors for breast cancer and asked to estimate her probabilities of developing breast cancer and dying of it within 10 years, with and without screening. The women's answers were compared with individual probabilities derived from the Gail et al. model, age-specific probabilities of developing and dying of breast cancer in the United States, and the results of randomized clinical trials of screening. RESULTS: The mailed questionnaire was completed and returned by 145 (73%) of the 200 women. Respondents over-estimated their probability of dying of breast cancer within 10 years by more than 20-fold (median, 22.3; interquartile range, 11.1-74.2). Assuming a 10% relative risk reduction from screening, respondents overestimated the relative risk reduction by sixfold (median, 6.0; interquartile range, 5.0-7.5) and the absolute risk reduction more than 100-fold (median, 127.5; interquartile range, 47.1-399.6). The median perceived estimate of absolute risk reduction was 6.0 breast cancer deaths per 100 women; the median calculated estimate was only 0.04 per 100 women. CONCLUSION: These findings suggest that many women younger than 50 years of age substantially overestimate their breast cancer risk and the effectiveness of screening. IMPLICATIONS: A balanced presentation of information about breast cancer risk and screening effectiveness may improve decision making for women younger than 50 years of age and reduce their anxiety about breast cancer, regardless of whether they choose to be screened.

Adult

Typing of LaCrosse, snowshoe hare, and Tahyna viruses by analyses of single-strand conformation polymorphisms of the small RNA segments.

Single-strand conformation polymorphism analysis was developed to differentiate the small RNA segments of three California serogroup bunyaviruses. The small RNA segments of La Crosse, snowshoe hare, and Tahyna viruses were reverse transcribed and PCR amplified. The cDNAs were then denatured, rapidly chilled to promote intrastrand reassociation, separated electrophoretically on a nondenaturing gel at room temperature, and silver stained. The resulting single-strand conformation polymorphism patterns were specific for the respective viruses. This molecular technique offers great potential for virus typing and taxonomic studies.

Base Sequence

Cost-effectiveness of head CT in patients with lung cancer without clinical evidence of metastases.

OBJECTIVE: To estimate the cost-effectiveness of CT for detecting brain lesions in patients with lung cancer without clinical evidence of metastases. DESIGN: Decision analysis model comparing two different strategies for detecting brain metastases: brain CT routinely (CT-first) or brain CT only when patients develop neurologic signs and/or symptoms (CT-deferred). PATIENTS: Hypothetical cohort of patients with lung cancer with an unremarkable screening clinical evaluation for metastases. MEASUREMENTS: Net costs are calculated as the difference in costs between the two limbs of the decision tree. Net benefits are expressed as the difference in calculated years of life expectancy between the two strategies. Net costs are divided by net benefits, yielding the marginal cost per quality adjusted year of added life expectancy (C/QALY) for the CT-first strategy. RESULTS: In the baseline analysis, the C/QALY for the CT-first strategy is about $70,000. Improving the clinical evaluation as a screen for detecting brain metastases markedly increases the C/QALY. Increasing the cost of brain CT magnifies this effect. More effective treatment for asymptomatic brain metastases and better accuracy of CT for identifying resectable and unresectable brain metastases lower C/QALY. CONCLUSIONS: Although a threshold cost-effectiveness has not been defined for identifying "cost-effective" diagnostic procedures, the marginal C/QALY of the CT-first strategy is substantially higher than many accepted medical interventions. At current costs, the routine use of brain CT is not warranted in patients with lung cancer who have normal findings on a standardized clinical evaluation for metastases.

Brain Neoplasms

Variation in ribosomal DNA intergenic spacers among populations of Anopheles albimanus in South and Central America.

Variation in the length and copy number of intergenic spacers (IGS) of nuclear ribosomal DNA were examined to test for genetic differentiation among Anopheles albimanus populations. Extensive collections were made in Guatemala but populations were also sampled over a large range of its distribution in Central and South America. Discriminant analysis of IGS patterns in individual mosquitoes indicated that populations generally had unique sets of IGS length variants. The IGS patterns from populations on the Pacific side of Central America were distinct from those on the Atlantic side or from South America. Cluster analysis indicated a similar trend. The IGS diversity in Central America was 50% greater than in South America. These results suggest that barriers to gene flow exist among Atlantic and Pacific coast populations of An. albimanus. No gene flow barriers were detected among populations from Colombia, Ecuador, Peru, and Venezuela.

Animals

Phylogeny of hard- and soft-tick taxa (Acari: Ixodida) based on mitochondrial 16S rDNA sequences.

Ticks are parasitiform mites that are obligate hematophagous ectoparasites of amphibians, reptiles, birds, and mammals. A phylogeny for tick families, subfamilies, and genera has been described based on morphological characters, life histories, and host associations. To test the existing phylogeny, we sequenced approximately 460 bp from the 3' end of the mitochondrial 16S rRNA gene (rDNA) in 36 hard- and soft-tick species; a mesostigmatid mite, Dermanyssus gallinae, was used as an outgroup. Phylogenies derived using distance, maximum-parsimony, or maximum-likelihood methods were congruent. The existing phylogeny was largely supported with four exceptions. In hard ticks (Ixodidae), members of Haemaphysalinae were monophyletic with the primitive Amblyomminae and members of Hyalomminae grouped within the Rhipicephalinae. In soft ticks (Argasidae), the derived phylogeny failed to support a monophyletic relationship among members of Ornithodorinae and supported placement of Argasinae as basal to the Ixodidae, suggesting that hard ticks may have originated from an Argas-like ancestor. Because most Argas species are obligate bird octoparasites, this result supports earlier suggestions that hard ticks did not evolve until the late Cretaceous.

Animals