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At least 325 records · Page 18Linked to original sources

A rapid manual method for CD4+ T-cell quantitation for use in developing countries.

OBJECTIVE: To evaluate a manual method (Cytosphere) for quantifying CD4+ T-cell numbers. DESIGN: Cross-sectional study of HIV-1-seronegative and HIV-1-seropositive individuals evaluated for absolute CD4 counts by both standardized flow cytometric measurements and manual Cytosphere technology using a hemacytometer. SETTING: University research hospitals in both the United States and Africa. PATIENTS, PARTICIPANTS: Blood specimens from 382 patients were evaluated. These were broken down into 294 samples obtained from HIV-1-seropositive patients and 88 samples obtained from HIV-1-seronegative patients. INTERVENTIONS: None. OUTCOME MEASURED: Absolute CD4 cell number. RESULTS: Evaluation of samples obtained from HIV-1 patients in both the United States and Africa demonstrated an overall correlation of the Cytosphere assay with flow cytometry of 0.912 (95% confidence interval, 0.895-0.928; P < 0.001). When samples were stratified based on CD4+ T-cell counts determined by flow cytometry, the Cytosphere assay had a 96% predictive value for correctly identifying individuals with CD4 T-cell counts > 200 x 10(6)/l and a 92% predictive value for correctly identifying individuals with CD4 T-cell counts < 200 x 10(6)/l. CONCLUSIONS: This assay appears to have the potential for the quantitation of CD4 cells in the limited laboratory facilities in developing countries and to have a strong correlation with standard flow cytometric technology.

Adult↗

Factors associated with active, refractory epistaxis.

This study addresses the underlying causes responsible for the severity and persistence of active, refractory epistaxis. Seventy-five patients referred because of treatment failure by primary care physicians showed hypertension and aspirin and alcohol abuse to be major factors in the refractory nature of their epistaxis. The majority of bleeding was located near the posterior floor of the nasal cavity and just posterior to Kiesselbach's plexus and was only associated with septal deviation, spurring, or mucosal abnormalities in 16 of the 75 patients. Seventeen of 67 outpatients required hospitalization. Standard laboratory tests were often inadequate determinants of etiology. Intractable epistaxis should be a signal for a thorough investigation of factors that influence clotting.

Anti-Inflammatory Agents, Non-Steroidal↗

Studies on the antibody response of mice and humans after immunization with potential influenza virus A (H1N1) vaccines.

The antibody response of mice and adult humans to immunization with subunit vaccines derived from a pair of antigenically distinct influenza A H1N1 viruses isolated in eggs was investigated. Although the haemagglutinin molecule of each virus differed by only three amino acid residues, highly specific antibody responses were elicited in mice as determined by haemagglutination inhibition and radioimmunoprecipitation assays. Results from competitive radioimmunoassays using monoclonal antibodies of known specificity and a study of the reactivity of mouse antisera with H1N1 field strains indicated that the marked differences in the antibody responses to the two vaccines was due to an amino acid substitution in the distal tip of the haemagglutinin molecule. In contrast, crossreactive antibody responses were elicited in humans presumably due to exposure to viruses related to the candidate vaccine prior to vaccination. Although immunogenic differences are apparent in this pair of antigenically distinct viruses in naive laboratory animals, these differences are not apparent following vaccination of humans that had prior exposure to related viruses.

Amino Acid Sequence↗

A new microprocessor-controlled anaesthetic machine.

This paper describes the development of a microprocessor controlled anaesthetic machine comprising an integrated anaesthetic apparatus and monitoring system. Following prolonged reliability trials in the laboratory, changes have been made to major components which were described in earlier publications.

Anesthesiology↗

Improving the outcomes of anticoagulation in rural Australia: an evaluation of pharmacist-assisted monitoring of warfarin therapy.

OBJECTIVE: The aim of this project was to assess whether rural pharmacist involvement in the management of patients receiving warfarin has the potential to lead to safer and more effective anticoagulation, and is valued and welcomed by patients and their general practitioners (GPs). METHODS: A convenience sample of rural pharmacists was trained in the use of the CoaguChek S International Normalized Ratio (INR) monitor and then conducted pharmacy-based testing for approximately 3 months. Two types of testing were performed in the pharmacy: (i) comparison testing was defined as pharmacy-based tests taken within 4 h of conventional laboratory testing or (ii) additional testing, which was a pharmacy-based test with no direct comparison laboratory test taken. Pharmacists, GPs and patients completed anonymous satisfaction surveys after the completion of the pharmacy-based testing. RESULTS: Pharmacists from 16 rural pharmacies were trained to use the CoaguChek S monitor. During the trial period, 518 INR tests were performed in the pharmacies on 137 different patients. A total of 120 tests were evaluated against results from laboratory testing. The pharmacy-based INR values were significantly correlated with the laboratory INR values (mean of 2.32+/-0.77 and 2.32+/-0.59 respectively; r=0.88, P<0.0001). A total of 398 additional pharmacy-based tests were conducted in the pharmacy and 8.5% of the additional tests resulted in a subsequent dosage change. The monitoring was well received by pharmacists, GPs and patients. CONCLUSIONS: The results of the trial were very positive. The CoaguChek S monitor in pharmacy-based testing performed accurately compared with conventional laboratory testing. Further research needs to be conducted on the impact of community pharmacy-conducted INR monitoring on patient care and outcomes.

Adult↗

Performance characteristics of the QUANTIPLEX HIV-1 RNA 3.0 assay for detection and quantitation of human immunodeficiency virus type 1 RNA in plasma.

The QUANTIPLEX HIV-1 RNA assay, version 3.0 (a branched DNA, version 3.0, assay [bDNA 3.0 assay]), was evaluated by analyzing spiked and clinical plasma samples and was compared with the AMPLICOR HIV-1 MONITOR Ultrasensitive (ultrasensitive reverse transcription-PCR [US-RT-PCR]) method. A panel of spiked plasma samples that contained 0 to 750,000 copies of human immunodeficiency virus type 1 (HIV-1) RNA per ml was tested four times in each of four laboratories (1,344 assays). Negative results (<50 copies/ml) were obtained in 30 of 32 (94%) assays with seronegative samples, 66 of 128 (52%) assays with HIV-1 RNA at 50 copies/ml, and 5 of 128 (4%) assays with HIV-1 RNA at 100 copies/ml. The assay was linear from 100 to 500,000 copies/ml. The within-run standard deviation (SD) of the log(10) estimated HIV-1 RNA concentration was 0.08 at 1,000 to 500,000 copies/ml, increased below 1,000 copies/ml, and was 0.17 at 100 copies/ml. Between-run reproducibility at 100 to 500 copies/ml was <0.10 log(10) in most comparisons. Interlaboratory differences across runs were </=0.10 log(10) at all concentrations examined. A subset of the panel (25 to 500 copies/ml) was also analyzed by the US-RT-PCR assay. The within-run SD varied inversely with the log(10) HIV-1 RNA concentration but was higher than the SD for the bDNA 3.0 assay at all concentrations. Log-log regression analysis indicated that the two methods produced very similar estimates at 100 to 500 copies/ml. In parallel testing of clinical specimens with low HIV-1 RNA levels, 80 plasma samples with <50 copies/ml by the US-RT-PCR assay had <50 copies/ml when they were retested by the bDNA 3.0 assay. In contrast, 11 of 78 (14%) plasma samples with <50 copies/ml by the bDNA 3.0 assay had >/=50 copies/ml when they were retested by the US-RT-PCR assay (median, 86 copies/ml; range, 50 to 217 copies/ml). Estimation of bDNA 3.0 values of <50 copies/ml by extending the standard curve of the assay showed that these samples with discrepant results had higher HIV-1 RNA levels than the samples with concordant results (median, 34 versus 17 copies/ml; P = 0.0051 by the Wilcoxon two-sample test). The excellent reproducibility, broad linear range, and good sensitivity of the bDNA 3.0 assay make it a very attractive method for quantitation of HIV-1 RNA levels in plasma.

DNA Probes↗

Quality control in the flow cytometric measurement of T-lymphocyte subsets: the multicenter AIDS cohort study experience. The Multicenter AIDS Cohort Study Group.

Since 1984, the Multicenter AIDS Cohort Study (MACS) has utilized four flow cytometry laboratories to measure T-lymphocyte subset levels semiannually in a large cohort of homosexual men. This report summarizes the steps taken in the MACS laboratories to attain comparability of lymphocyte subset determinations across the centers and over time. Identical flow cytometers, monoclonal antibodies, and analytic procedures have been used, and over a period of time, the procedure for sample preparation was also standardized. Interlaboratory proficiency testing utilizing identical specimens analyzed in the four laboratories was performed to evaluate the comparability of the data among the laboratories. Our results verify that such testing can identify technical bias in flow cytometric evaluations performed at different laboratories. Temporal laboratory consistency in flow cytometric measurements was evaluated using data from each site's HIV-seronegative homosexual reference group. Both sequential 95% confidence intervals (mean +/- 2 x SEM) and the within-person standard deviations of the immune measurements were considered. Significant variation in CD3, CD4, and CD8 lymphocyte subset percentages over time in the seronegative reference population was observed. Our observations indicate that the lymphocyte subset values of this seronegative group should be used to adjust those obtained on the seropositive study participants during a particular time period, thereby allowing improved discrimination of the effects of HIV on T cells in infected individuals. The data presented are of use for designing epidemiologic and intervention studies in HIV-1-infected individuals, especially for calculating sample sizes. The methods we have used to assess the quality of data in the MACS have general application to quality control programs in flow cytometry laboratories. In particular, comparison of sequential confidence intervals and within-person standard deviations for lymphocyte subset determinations on control populations are essential to a comprehensive proficiency testing program because they permit assessment of consistency within a laboratory over time.

Acquired Immunodeficiency Syndrome↗

Comparison of polymerase chain reaction with other laboratory methods for the diagnosis of American cutaneous leishmaniasis: diagnosis of cutaneous leishmaniasis by polymerase chain reaction.

An evaluation of 5 laboratory methods for diagnosing American cutaneous leishmaniasis (ACL) was carried out on patients from an endemic area of Brazil. From 164 patients presenting cutaneous lesions, and suspected to have ACL, 133 (81.1%) were confirmed for the disease by Montenegro skin test (MST) and/or parasitologic examination (PE). In both groups of patients, the positivity of polymerase chain reaction (PCR) was similar to that of immunofluorescence assay and enzyme-linked immunosorbent assay, and higher than that of MST and PE (P < .05). In the group of patients suspected to have ACL, PCR presented the same positivity as PE and MST together. No correlation between positivity of the laboratory methods and clinical or epidemiologic aspects was observed. Our data confirmed the value of PCR as an alternative laboratory method for diagnosing ACL, especially for those patients with negative PE and MST.

Animals↗

Genetic characterization of Pseudomonas fluorescens SBW25 rsp gene expression in the phytosphere and in vitro.

The plant-colonizing Pseudomonas fluorescens strain SBW25 harbors a gene cluster (rsp) whose products show similarity to type III protein secretion systems found in plant and animal pathogens. Here we report a detailed analysis of the expression and regulation of the P. fluorescens rsp pathway, both in the phytosphere and in vitro. A combination of chromosomally integrated transcriptional reporter fusions, overexpressed regulatory genes, and specific mutants reveal that promoters controlling expression of rsp are actively transcribed in the plant rhizosphere but not (with the exception of the rspC promoter) in the phyllosphere. In synthetic medium, regulatory (rspL and rspR) and structural (rspU, plus the putative effector ropE) genes are poorly expressed; the rspC promoter is subject to an additional level of regulatory control. Ectopic expression of regulatory genes in wild-type and mutant backgrounds showed that RspR controls transcription of the alternate sigma factor, rspL, and that RspL controls expression of gene clusters encoding structural genes. Mutation of rspV did not affect RspR-mediated expression of rspU. A search for additional regulators revealed two candidates--one with a role in the conversion of alanine to pyruvate--suggesting that expression of rsp is partly dependent upon the metabolic status of the cell. Mutations in rsp regulators resulted in a significant reduction in competitive colonization of the root tips of sugar beet seedlings but also caused a marked increase in the lag phase of laboratory-grown cultures, indicating that rsp regulatory genes play a more significant general role in the function of P. fluorescens SBW25 than previously appreciated.

Arabidopsis↗

Acute flaccid paralysis and West Nile virus infection.

Acute weakness associated with West Nile virus (WNV) infection has previously been attributed to a peripheral demyelinating process (Guillain-Barré syndrome); however, the exact etiology of this acute flaccid paralysis has not been systematically assessed. To thoroughly describe the clinical, laboratory, and electrodiagnostic features of this paralysis syndrome, we evaluated acute flaccid paralysis that developed in seven patients in the setting of acute WNV infection, consecutively identified in four hospitals in St. Tammany Parish and New Orleans, Louisiana, and Jackson, Mississippi. All patients had acute onset of asymmetric weakness and areflexia but no sensory abnormalities. Clinical and electrodiagnostic data suggested the involvement of spinal anterior horn cells, resulting in a poliomyelitis-like syndrome. In areas in which transmission is occurring, WNV infection should be considered in patients with acute flaccid paralysis. Recognition that such weakness may be of spinal origin may prevent inappropriate treatment and diagnostic testing.

Adult↗

Blockage of progesterone-induced release of LH by intrabrain implants of actinomycin D.

Implants of cocoa butter (CB) or CB and actinomycin D (AD) were placed in the hypothalamus, amygdala, or anterior pituitary gland of testosterone-pretreated ovariectomized rats. The animals were then given an injection of 2.5 mg progesterone (P) at 11.00 h and sacriviced at approximately 16.30 h. Serum LH was determined by radioimmunoassay. Implants of AD placed in or near the arcuate nucleus and in other sites in the ventromedial hypothalamus blocked p-induced LH release, but those placed in the preoptic area or ventrolateral hypothalamus were only partially effective. Implants of AD in the amygdala or anterior pituitary gland had no detectable effect on LH release. Approximately 70 percent of the radioactivity in implants of 3H-AD diffused from the implant site within 8 h. of that recovered, most was found within 1 mm of the implant site. These results suggest that the ventromedial hypothalamus is one site at which AD acts to block P-induced LH release.

Amygdala↗

Quality assurance and use of PCR in clinical trials.

Qualitative and quantitative HIV-1 DNA and RNA PCR assays are proving to be useful in the diagnosis of HIV-1 infection in infants and in assessing the in vivo antiviral activity of new therapies and regimens in clinical trials. The use of these standardized commercial assays in conjunction with an external quality assurance program has ensured that results from different laboratories are comparable. In addition, real-time proficiency monitoring has the potential to detect problems immediately before patient data are compromised.

Clinical Trials as Topic↗

Pathways to diagnosis of cervical cancer: screening history, delay in follow up, and smear reading.

BACKGROUND: The aim of this study was to determine the most important ways to reduce incidence of and mortality from cervical cancer by a nationally co-ordinated screening programme. DESIGN: Descriptive study. SETTING: The New Zealand National Cervical Screening Programme: a nationally organised and co-ordinated programme. SAMPLE: Women aged younger than 80 years with histologically proven primary invasive cervical cancer, including microinvasive disease, diagnosed between 1 January 2000 and 30 September 2002. Consent for access to medical records was gained for 371 of 445 eligible women (83%). A total of 359 (81%) of eligible women or their next of kin consented to interview. METHODS: Data on events prior to diagnosis were obtained from routine sources, interview, medical record review and slide reread. MAIN OUTCOME MEASURES: Frequency of screening in the 7 years prior to diagnosis, time from abnormal smear or symptoms to appropriate diagnostic confirmation, proportion of negative smears upgraded to high grade on reread. RESULTS: Half of the 371 participants (83% of 445 eligible women) had not had a screening smear in the 3 years prior to diagnosis, and 80% were defined as inadequately screened. A maximum of 17% of women overall or within any defined subgroup experienced delays in follow up of abnormal smears or bleeding. Only 11% of women overall had had a high-grade smear, which was originally read as negative. CONCLUSIONS: The most important factor in women's pathways to a diagnosis of cervical cancer was inadequate screening. While delays in diagnosis could be reduced and laboratory performance improved, priority must be given to improving uptake and frequency of screening.

Adult↗

Oral ribavirin treatment of influenza A and B.

A loading dose and short-term administration of oral ribavirin significantly improved symptoms and signs of influenza type A or B infection in 25 patients. The antiviral effect was not significant. No adverse clinical effects or significant laboratory values were observed. Oral treatment of patients with influenza A or B infection might be possible with ribavirin.

Administration, Oral↗

HIV-1 plasma RNA level and CD4 cell count in a large urban HIV-1-infected patient population from 1997 to 2000.

We evaluated the change in plasma HIV-1 RNA level and CD4 cell counts in an HIV-1-infected population between 1997 and 2000. Both the mean and the median values of plasma HIV-1 RNA level decreased with time with the exception of 1998. The mean and medians for CD4 cell count appear to be fairly stable. While the percentage of patients with plasma HIV-1 RNA level <400 copies/mL increased from 29.12% in 1997 to 41.31% in 2000, the percentage of patients with plasma HIV-1 RNA level >100,000 copies/mL remained fairly constant. The availability of HAART had impacted the level of plasma HIV-1 RNA, although many patients still have plasma HIV-1 RNA level >100,000 copies/mL. Most patients with plasma HIV-1 RNA level >400 copies/mL in 1997 still have plasma HIV-1 RNA level >400 copies/mL in 2000. These laboratory findings, however, do not necessarily mean that there had been a lack of clinical benefit.

Antiretroviral Therapy, Highly Active↗

Validity of hair mineral testing.

The variance of testing was compared between the College of American Pathologists clinical survey and that of a recent review about hair mineral testing. The review suggested that the accuracy of hair mineral testing was unreliable. In general, there was a greater range of variance in the College of American Pathologists testing results. These latter results are based on laboratory testing and are used as a "yardstick" to determine if a laboratory passes or fails that analyte and are considered a "gold standard." An extract, which resulted from a method that avoided the washing step, was compared among five laboratories. Very good precision resulted, indicating that the varied washing steps used by the laboratories in a recent review were probably the source of much variance. Analysis of hair analysis seemed to yield important information in several historical or forensic cases involving Ludwig von Beethoven, Napoleon Bonaparte, ex-US-presidents Zachary Taylor and Andrew Jackson, and Charles Hall, an Arctic explorer. Several elements that were reviewed, including arsenic, cadmium, cobalt, germanium, lead, lithium, manganese, mercury, nickel, and thallium, showed relationships between body burden, dosage, and exposure or toxicity. Evidence of toxicity could not be found by measuring hair aluminum or vanadium. Chromium, selenium, and zinc seemed to have nutritional value. Ratios of hair elements with clinical importance could not be found.

Famous Persons↗