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Variable selection and multivariate methods for the identification of microorganisms by flow cytometry.

BACKGROUND: When exploited fully, flow cytometry can be used to provide multiparametric data for each cell in the sample of interest. While this makes flow cytometry a powerful technique for discriminating between different cell types, the data can be difficult to interpret. Traditionally, dual-parameter plots are used to visualize flow cytometric data, and for a data set consisting of seven parameters, one should examine 21 of these plots. A more efficient method is to reduce the dimensionality of the data (e.g., using unsupervised methods such as principal components analysis) so that fewer graphs need to be examined, or to use supervised multivariate data analysis methods to give a prediction of the identity of the analyzed particles. MATERIALS AND METHODS: We collected multiparametric data sets for microbiological samples stained with six cocktails of fluorescent stains. Multivariate data analysis methods were explored as a means of microbial detection and identification. RESULTS: We show that while all cocktails and all methods gave good accuracy of predictions (>94%), careful selection of both the stains and the analysis method could improve this figure (to > 99% accuracy), even in a data set that was not used in the formation of the supervised multivariate calibration model. CONCLUSIONS: Flow cytometry provides a rapid method of obtaining multiparametric data for distinguishing between microorganisms. Multivariate data analysis methods have an important role to play in extracting the information from the data obtained. Artificial neural networks proved to be the most suitable method of data analysis.

Bacillus subtilis↗

Survival in patients with pulmonary arterial hypertension treated with first-line bosentan.

BACKGROUND: Pulmonary arterial hypertension (PAH) is a devastating disease of the small pulmonary arteries and arterioles, characterized by intimal fibrosis, medial hypertrophy and plexiform lesions. When untreated both the idiopathic form (IPAH, formerly termed primary pulmonary hypertension, PPH) and PAH related to various other conditions such as scleroderma (SSc) often take a progressive course with high mortality. There is ongoing search for disease-specific treatments that are able to improve survival in these patients. The oral dual endothelin (ET(A)/ET(B)) antagonist bosentan has been shown to improve exercise capacity, time to clinical worsening, haemodynamics and quality of life in short-term studies. MATERIALS AND METHODS: To determine the long-term effects of bosentan on survival, patients from the two double-blind, randomized trials and their open-label extensions, treated with first-line bosentan, were followed for up to 3 years. Data on survival were collected between September 1999 (first patient included in the placebo-controlled trials) and December 2002. Vital status was verified in each patient. The survival cohorts of these patients were compared with either the predicted survival for each patient based on an equation from the National Institutes of Health (NIH) PPH registry or with historical controls. RESULTS: Observed survival up to 36 months was reported as a Kaplan-Meier estimate in three cohorts: (1) In 169 PPH patients treated with first-line bosentan, 1- and 2-year survival was 96% and 89%, respectively, vs. predicted untreated survival at 1 and 2 years of 69% and 57%, respectively; (2) in 50 patients with PAH associated with SSc (PAH-SSc), 1-, 2- and 3-year survival was 82%, 67% and 64%, respectively, vs. approximately 45%, approximately 35% and approximately 28%, respectively, from registry data of untreated PAH-SSc patients; and (3) in 139 PPH patients in WHO functional class III, 1- and 2-year survival was 97% and 91%, respectively, vs. 91% and 84% in a historical cohort of 346 patients treated with epoprostenol in five major referral centres. CONCLUSIONS: The present analyses suggest that first-line bosentan therapy, followed by the addition of other disease-specific therapies as required, improves survival in patients with advanced PAH.

Adolescent↗

[Soft tissue changes in adults with acquired growth hormone deficiency during substitution treatment. A double-blind, randomized, placebo-controlled study after a year of treatment].

The aim of the study was to assess the influence of growth hormone (GH) on body composition in 29 adult persons with acquired growth hormone deficiency (GHD). The study was a double-blind placebo-controlled study of one year's duration. The treatment consisted of daily subcutaneous injections of GH (2.0 IU/m2, Norditropin) or placebo. Before and after one year's treatment, the patients were hospitalized for measurement of Dual-Energy X-ray Absorptiometry (DXA) in order to assess body composition (Fat mass (FM) + Lean Body Mass (LBM). In addition, fasting serum Insulin-like Growth Factor (IGF-I) was collected. The study showed a 200% increase in serum IGF-I. Regarding LBM a 5.7% increase was seen, and this was primarily due to an increase in LBM of the extremities, whereas FM was reduced by 21.5% mainly located truncally. Our data demonstrate that favorable changes in body composition are maintained during long-term, controlled GH treatment, which loads support to the concept of GH substitution in GH-deficient adults.

Absorptiometry, Photon↗

Dual-color total internal reflection fluorescence cross-correlation spectroscopy.

We present the development and first application of a novel dual-color total internal reflection (TIR) fluorescence system for single-molecule coincidence analysis and fluorescence cross-correlation spectroscopy (FCCS). As a performance analysis, we measured a synthetic DNA-binding assay, demonstrating this dual-color TIR-FCCS approach to be a suitable method for measuring coincidence assays such as biochemical binding, fusion, or signal transduction at solid/liquid interfaces. Due to the very high numerical aperture of the epi-illumination configuration, our setup provides a very high fluorescence collection efficiency resulting in a two- to three-fold increase in molecular brightness compared to conventional confocal FCCS. Further improvements have been achieved through global analysis of the spectroscopic data.

DNA↗

Effects of loss of classical estrogen response element signaling on bone in male mice.

The role of estrogen signaling in the male skeleton via estrogen receptor (ER)-alpha is now well established. ERalpha can elicit responses through either classical estrogen response elements (ERE) pathways or nonclassical, non-ERE pathways. In the present study, we examined the effects of either the attenuation or loss of classical ERalpha signaling on the murine male skeleton. To accomplish this, we crossed male mice heterozygous for a knock-in mutation [nonclassical ERalpha knock-in (NERKI)], which abolishes the ERE-mediated pathway with female heterozygous ERalpha knockout mice (ERalpha+/-) and studied the F1 generation ERalpha+/+, ERalpha+/-, ERalpha+/NERKI, and ERalpha-/NERKI male progeny longitudinally using bone density and histomorphometry. The only ERalpha allele present in ERalpha-/NERKI mice is incapable of classical ERE-mediated signaling, whereas the heterozygous ERalpha+/NERKI mice have both one intact ERalpha and one NERKI allele. As compared with ERalpha+/+ littermates (n=10/genotype), male ERalpha+/NERKI and ERalpha-/NERKI mice displayed axial and appendicular skeletal osteopenia at 6, 12, 20, and 25 wk of age, as demonstrated by significant reductions in total bone mineral density (BMD) at representative sites (areal BMD by dual-energy x-ray absorptiometry at the lumbar vertebrae and femur and volumetric BMD by peripheral quantitative computed tomography at the tibia; P<0.05-0.001 vs. ERalpha+/+). The observed osteopenia in these mice was evident in both trabecular and cortical bone compartments. However, these decreases were more severe in mice lacking classical ERalpha signaling (ERalpha-/NERKI mice), compared with mice in which one wild-type ERalpha allele was present (ERalpha+/NERKI mice). Collectively, these data demonstrate that classical ERalpha signaling is crucial for the development of the murine male skeleton.

Animals↗

Man and polar bear in Svalbard: a solvable ecological conflict?

The objective of this study is twofold. First, it is to assess the nature and magnitude of the polar bear-human conflict with respect to injuries to man and bear. Second, a major concern has been to minimize injurious interactions in order to safeguard the people who live and work in the Arctic, and, at the same time, secure the future of the polar bear in one of the last relatively unspoiled habitats on earth for big carnivores. From 1971 to 1995, approximately 80 bears were involved in serious bear-human interactions. Of these, 77 bears were killed and 3 escaped after having injured people. During the same period, 10 people were injured, 4 of them fatally, in 7 separate interactions, each involving a single bear. None of the victims carried an appropriate firearm. The circumstances leading up to the confrontations give strong reasons for supposing that the majority of the attacks were predatory in nature. Seven of the injured, including the four who were killed, sustained bites to the head and neck. Correct use of firearms could probably have prevented all the fatalities. However, the keeping and use of firearms caused two accidental deaths in the same period. We conclude that alertness, the absence of attractants (food, garbage), and appropriate bear repellents to secure field camps are important items in preventing conflicts and should always be available. However, as a last but indispensable resort, a firearm (rifle or shotgun) carried by an experienced user is the only safe precaution for avoiding injuries in polar bear country. Killing a bear on the rare occasions when humans are in danger presents no threat to the bear population. With regard to physical injury to people, the problem is a minor one. Bears have a dual impact on everyday life in the Svalbard settlements. While there is some anxiety related to the presence of bears, the polar bear is a source of breathtaking adventure highly valued by both residents and visitors.

Adult↗

MKP-3 has essential roles as a negative regulator of the Ras/mitogen-activated protein kinase pathway during Drosophila development.

Mitogen-activated protein kinase (MAPK) phosphatase 3 (MKP-3) is a well-known negative regulator in the Ras/extracellular signal-regulated kinase (ERK)-MAPK signaling pathway responsible for cell fate determination and proliferation during development. However, the physiological roles of MKP-3 and the mechanism by which MKP-3 regulates Ras/Drosophila ERK (DERK) signaling in vivo have not been determined. Here, we demonstrated that Drosophila MKP-3 (DMKP-3) is critically involved in cell differentiation, proliferation, and gene expression by suppressing the Ras/DERK pathway, specifically binding to DERK via the N-terminal ERK-binding domain of DMKP-3. Overexpression of DMKP-3 reduced the number of photoreceptor cells and inhibited wing vein differentiation. Conversely, DMKP-3 hypomorphic mutants exhibited extra photoreceptor cells and wing veins, and its null mutants showed striking phenotypes, such as embryonic lethality and severe defects in oogenesis. All of these phenotypes were highly similar to those of the gain-of-function mutants of DERK/rl. The functional interaction between DMKP-3 and the Ras/DERK pathway was further confirmed by genetic interactions between DMKP-3 loss-of-function mutants or overexpressing transgenic flies and various mutants of the Ras/DERK pathway. Collectively, these data provide the direct evidences that DMKP-3 is indispensable to the regulation of DERK signaling activity during Drosophila development.

Animals↗

Frequency of osteoporosis in 187 men with rheumatoid arthritis followed in a university hospital.

OBJECTIVE: Although there is relevant information on frequency of osteoporosis in women with rheumatoid arthritis (RA), data about male patients are limited. We evaluated the frequency of osteoporosis in a group of Spanish men with RA followed in a university hospital. METHODS: From the database of our bone densitometry unit, we searched for men with RA evaluated between January 1991 and December 2004 and identified 187 patients, 156 of whom were older than 50 years. Previously recorded demographic, disease, and treatment-related variables were collected. Bone mineral density (BMD) was measured by dual x-ray absorptiometry (DEXA). Osteoporosis was defined according to the criteria of the World Health Organization (WHO), recommended for postmenopausal Caucasian women, as a T score <or= -2.5 SD, and the International Society for Clinical Densitometry (ISCD), which indicates the same diagnostic threshold, but only in men over age 50 years. RESULTS: BMD was lower than in the general population in both lumbar spine [Z score -0.41 +/- 1.00 (95% CI -0.55 to -0.26)] and femoral neck [Z score -0.46 +/- 0.95 (95% CI -0.60 to -0.31)]. When the WHO threshold for postmenopausal women was applied, frequency of osteoporosis was 13% at lumbar spine, 12% at the femoral neck, and 21% in at least one of the evaluated sites. When ISCD criteria were applied, the frequency of osteoporosis was 13%, 14%, and 23%, respectively. CONCLUSION: Frequency of osteoporosis in men is considerably lower than that traditionally established in women with RA, independent of the diagnostic criteria applied.

Absorptiometry, Photon↗

Epidemiology of Diabetes Interventions and Complications (EDIC). Design, implementation, and preliminary results of a long-term follow-up of the Diabetes Control and Complications Trial cohort.

OBJECTIVE: The Diabetes Control and Complications Trial (DCCT) demonstrated the powerful impact of glycemic control on the early manifestations of microvascular complications. Contemporary prospective data on the evolution of macrovascular and late microvascular complications of type 1 diabetes are limited. The Epidemiology of Diabetes Interventions and Complications (EDIC) study is a multicenter, longitudinal, observational study designed to use the well-characterized DCCT cohort of > 1,400 patients to determine the long-term effects of prior separation of glycemic levels on micro- and macrovascular outcomes. RESEARCH DESIGN AND METHODS: Using a standardized annual history and physical examination, 28 EDIC clinical centers that were DCCT clinics will follow the EDIC cohort for 10 years. Annual evaluation also includes resting electrocardiogram. Doppler ultrasound measurements of ankle/arm blood pressure, and screening for nephropathy. At regular intervals, a timed 4-h urine is collected, lipid profiles are obtained, and stereoscopic fundus photographs are taken. In addition, dual B-mode Doppler ultrasound scans of the common and internal carotid arteries will be performed at years 1 and 6 and at study end. RESULTS: Written informed consent was obtained from 96% of the DCCT subjects. The participants, compared with nonparticipants, tended to have better glycemic control at the completion of the DCCT and were more likely to have their diabetes care provided by DCCT personnel. The EDIC baseline measurement stratified by sex delineates multiple cardiovascular disease risk factor differences such as age (older in men), waist-to-hip ratio (higher in men). HDL cholesterol (lower in men), hypertension (more prevalent in men), and maximum intimal-medial thickness of common and internal carotid arteries (thicker in men). Of the original conventional treatment group, 69% have changed to continuous subcutaneous insulin infusion or multiple daily injections. Although the mean HbA1c difference between the intensive and conventional treatment groups narrowed at EDIC years 1 and 2, HbA1c remained significantly lower in the intensive group. Of all expected clinic visits, 95% were completed, and the quality of EDIC data is very similar to that observed in the DCCT. CONCLUSIONS: Although obvious problems exist in extended follow-up studies of completed clinical trials, these are balanced by the value of continued systematic observation of the DCCT cohort. In contrast to other epidemiologic studies, EDIC will provide 1) definitive data on type 1 as distinct from type 2 diabetes; 2) reliance on prospective rather than on cross-sectional analysis; 3) long-term follow-up in a large population; 4) consistent use of objective, reliable measures of outcomes and glycemia; and 5) observation of patients from before the onset of complications.

Adult↗

Television advertisement format and the provision of risk information about prescription drug products.

BACKGROUND: Considerable attention has been afforded to analyzing the content of and assessing consumers' reaction to print direct-to-consumer drug ads, but not so for televised ads. OBJECTIVE: To determine whether advertisements with different risk severity and risk presentation would significantly affect viewers' (1) recall of information contained in the advertisement, (2) evaluation of the advertisement, and (3) perceptions of the advertised product's risks. METHODS: Data were collected from a sample of 135 first-year pharmacy students at a Midwestern college of pharmacy. After viewing 1 of the 6 advertisements designed for this study, participants were asked to complete a self-administered survey. Chi-square and analysis of variance were used to analyze the data. A 2x3 between subjects design was used to test the effects of 2 levels of risk severity (high- vs low-risk severity) and 3 levels of risk presentation (original ad containing integrated risk message, deintegrated risk message/dual modality using male voice-over, deintegrated risk message/dual modality using female voice-over). RESULTS: Results of analysis of variance procedures revealed that deintegrating risk information by placing it at the end of the advertisement and the use of captions in addition to oral messages (dual modality) (1) improved the recall of general and specific side effect information, (2) led to a perception that the advertisement had greater informational content, (3) resulted in lower Advertisement Distraction, and (4) lessened cognitive and affective aspects of information overload for the advertisement containing the high-risk severity medication. However, this pattern of findings was not found for the low-risk severity medication. CONCLUSION: Alternative methods for presenting risk information in direct-to-consumer ads affected some aspects of information recall and advertisement evaluation, but were not shown to affect risk perceptions regarding the advertised products.

Adult↗

Dual natriuretic peptide response to volume load in the fetal circulation.

OBJECTIVE: To measure atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in control fetuses and fetuses with Rhesus isoimmunisation before and after intravascular transfusion. The current study was designed to investigate the response of ANP and BNP to cardiac short-term and long-term volume load in the human fetus. METHODS: Fetal blood samples were collected from 18 human fetuses (nine controls, nine anemic fetuses with Rhesus isoimmunisation before and after intravascular transfusion). Fetal ANP and BNP concentrations were measured and compared to maternal plasma levels. RESULTS: Both ANP and BNP were significantly higher in fetal blood compared to the mothers. Fetuses with Rhesus isoimmunisation, characterized by long-term cardiac overload, showed significantly elevated ANP but not BNP concentration compared to the fetal controls (ANP: 80.8+/-16.6 vs. 31.6+/-7.7 pg/ml, P<0.05). However, short-term volume load due to intravascular transfusion leads to a significant increase in the fetal BNP- but not ANP-plasma level (BNP: 112.9+/-14.1 vs. 64.8+/-6.6 pg/ml, P<0.05). CONCLUSION: ANP and BNP respond differently to cardiac short- and long-term volume load in the fetal circulation. Therefore, the data suggest that in the fetus, similar to adults, ANP and BNP constitute a dual natriuretic peptide system responsive to changes in cardiac filling pressure.

Analysis of Variance↗

Clinical decision rules in rheumatoid arthritis: do they identify patients at high risk for osteoporosis? Testing clinical criteria in a population based cohort of patients with rheumatoid arthritis recruited from the Oslo Rheumatoid Arthritis Register.

BACKGROUND: Preliminary clinical criteria based on age, inflammation, and immobility have been proposed to identify which patients with rheumatoid arthritis (RA) should be examined by dual energy x ray absorptiometry (DXA) to diagnose osteoporosis. The three item criteria have not been evaluated in male patients with RA or in the entire female RA population. OBJECTIVES: (1) To test the proposed criteria in a cohort of men and women thought to be representative of the entire underlying RA population. (2) To develop clinical decision rules, which could be applied to all patients with RA irrespective of corticosteroid use. METHODS: Clinical and demographic data were collected from a total of 287 representative patients with RA (235 (82%) women, 52 (18%) men, age range 25.3-73.1 years) from the Oslo RA register (completeness 85%). Bone mineral density (BMD) was measured in spine L2-4 (anterior-posterior view) and femoral neck by DXA. The criteria were applied and sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated. RESULTS: Mean age (SD) for the women and men with RA was 56.8 (11.0) years and 61.5 (10.2) years; disease duration was 15.5 (9.5) years and 14.7 (8.6) years. Of the women 163 (69%) were postmenopausal. One hundred and seventeen (50%) women and 28 (54%) men fulfilled the three item criteria. For the diagnosis of osteoporosis (T score <or=-2.5) using the original three item criteria sensitivity in women and men was 74% and 67%, specificity 57% and 50%, PPV 32% and 29% and NPV 89% and 83%, and including weight and ever use of corticosteroids in a five item criteria sensitivity increased to 82% and 83%, specificity decreased to 45% and 45%, PPV was 29% and 31%, and NPV was 90% and 90% respectively. CONCLUSION: The novel five item criteria (age, weight, inflammation, immobility, and ever use of corticosteroids) are a more accurate tool to identify patients with RA and osteoporosis than the original three item criteria (age, inflammation, and immobility). The clinical decision rules have an acceptable sensitivity and provide a practical tool for the doctor to identify patients with RA who should have a DXA measurement performed.

Absorptiometry, Photon↗

Schistosomiasis and HIV-1 infection in rural Zimbabwe: implications of coinfection for excretion of eggs.

BACKGROUND: Stunted development and reduced fecundity of Schistosoma parasites in immunodeficient mice and the impaired ability of human immunodeficiency virus 1 (HIV-1)-infected humans to excrete schistosome eggs have been described. This study explores the effect that HIV-1-associated immunodeficiency has on the excretion of schistosome eggs in a large cohort of coinfected individuals. METHODS: In a cross-sectional survey, urine and stool samples were obtained from and HIV-1 status was determined for 1545 individuals. More extensive data, including quantitative measures of intensity of infection in schistosomiasis and immunodeficiency, were collected in the Mupfure schistosomiasis and HIV longitudinal cohort, composed of 379 participants of whom 154 were coinfected with HIV-1 and Schistosoma parasites. RESULTS: In the cross-sectional survey, the overall prevalence of schistosomiasis was 43.4%, and 26.3% of the participants were infected with HIV-1. Schistosome infections were due to Schistosoma haematobium in 63.6% of cases, S. mansoni in 18.1% of cases, and dual infections in 18.4% of cases. Intensities of Schistosoma infections, measured by the number of eggs excreted and by the level of circulating anodic antigens, did not differ between HIV-1-negative and HIV-1-positive participants coinfected with S. haematobium, S. mansoni, or both. CD4 cell counts were significantly lower in HIV-1-positive participants and in S. mansoni-infected HIV-1-negative participants than in other participants. CONCLUSION: The present study suggests that adult HIV-1-related immunodeficiency does not impair the ability to excrete eggs in low-intensity infection with S. haematobium, S. mansoni, or both and that infection with HIV-1 may not have major implications for diagnosis and surveillance of schistosomiasis.

Adult↗

The barley ERF-type transcription factor HvRAF confers enhanced pathogen resistance and salt tolerance in Arabidopsis.

We isolated HvRAF (Hordeum vulgare root abundant factor), a cDNA encoding a novel ethylene response factor (ERF)-type transcription factor, from young seedlings of barley. In addition to the most highly conserved APETALA2/ERF DNA-binding domain, the encoded protein contained an N-terminal MCGGAIL signature sequence, a putative nuclear localization sequence, and a C-terminal acidic transcription activation domain containing a novel mammalian hemopexin domain signature-like sequence. Their homologous sequences were found in AAK92635 from rice and RAP2.2 from Arabidopsis; the ERF proteins most closely related to HvRAF, reflecting their functional importance. RNA blot analyses revealed that HvRAF transcripts were more abundant in roots than in leaves. HvRAF expression was induced in barley seedlings by various treatment regimes such as salicylic acid, ethephon, methyl jasmonate, cellulase, and methyl viologen. In a subcellular localization assay, the HvRAF-GFP fusion protein was targeted to the nucleus. The fusion protein of HvRAF with the GAL4 DNA-binding domain strongly activated transcription in yeast. Various deletion mutants of HvRAF indicated that the transactivating activity was localized to the acidic domain of the C-terminal region, and that the hemopexin domain signature-like sequence was important for the activity. Overexpression of the HvRAF gene in Arabidopsis plants induced the activation of various stress-responsive genes, including PDF1.2, JR3, PR1, PR5, KIN2, and GSH1. Furthermore, the transgenic Arabidopsis plants showed enhanced resistance to Ralstonia solanacearum strain GMI1000, as well as seed germination and root growth tolerance to high salinity. These results collectively indicate that HvRAF is a transcription factor that plays dual regulatory roles in response to biotic and abiotic stresses in plants.

Adaptation, Physiological↗

The World Survey of Cardiac Pacing and Implantable Cardioverter Defibrillators: calendar year 1997--Europe.

The registry of the European Working Group on Cardiac Pacing (EWGCP) is based on the European Pacemaker Identification Card originally designed in July 1978. National registration centers collect the local data and send aggregated annual data to the EWGCP. For 1997, data were obtained from 2,887 hospitals in 20 European countries representing a population of 568 million. Across all participating countries, the median value for all implanted pacemakers was 378 per million population. For initial pacemaker implants, the median value was 290 per million population. Single chamber atrial pacing was important in Denmark, the Netherlands, Poland, Slovak Republic, Spain, and Sweden for the treatment of sick sinus syndrome. Dual chamber pacing accounted for < 50% of initial implants in only 5 of 14 countries for atrioventricular block, and in only 3 of 15 countries for sick sinus syndrome. In 7 of 15 countries, unipolar ventricular leads were used in > or = 50% of cases. In 6 of 14 countries, there was > 15% use of unipolar atrial leads. Nine of 13 countries frequently used atrial active-fixation leads. For the 1997 survey, ICD data were obtained from 16 countries. The total number of ICDs per million population was a median value of 14. Initial ICD implants per million population was 11. Only 3 of 16 countries implanted a total of 30 or more ICDs per million population. Pacing and ICD practices were dependent on the availability of medical and technical resources and influenced by economic constraints inherent in health care administration and insurance coverage patterns.

Aged↗

An outbreak of Mayaro virus disease in Belterra, Brazil. III. Entomological and ecological studies.

Results in entomological and vertebrate host investigations made during dual outbreaks of Mayaro (MAY) and yellow fever (YF) viruses in Belterra, Pará, Brazil in 1978 are reported. Over 9,000 insects representing 26 species were assayed in 396 pools for the presence of arboviruses. Pools of Haemagogus janthinomys Dyar yielded the only isolates of either MAY or YF virus. The minimum field infection rate for nine isolates of MAY virus from Hg. janthinomys was 1:82, and for two isolates of YF virus was 1:368. Analysis of collection data showed Hg. janthinomys to be attracted to man as a blood source and present in all habitats sampled, although most abundant in the forest canopy. Twelve hundred bird sera and 584 mammal sera were tested by hemagglutination-inhibition (HI) tests for antibody to MAY virus. Highest MAY antibody prevalence rates were found among marmosets (Calithrix argentata, 32 positive of 119 tested, 27%). Mayaro virus was also isolated from the blood of a sylvan marmoset captured at the peak of the MAY virus outbreak. Experimental infection of marmosets with MAY virus confirmed that a substantial viremia follows infection with this virus. Marmosets were also found with HI antibody to YF virus (5/119, 4%). The results presented indicate that Hg. janthinomys was the principal vector of both MAY and YF viruses and that marmosets were the main amplifying hosts for MAY virus, and perhaps for YF virus as well.

Alphavirus↗

Dual-spectra ultrasonography: an attenuation-compensating technique for myocardial perfusion analysis.

OBJECTIVE: To validate the harmonic-fundamental frequency ratio peak for user-independent differentiation of myocardial perfusion under clinically relevant levels of signal attenuation. METHODS: Radio frequency data were obtained by using apical long-axis scans in 11 open-chest pigs during continuous infusion of a contrast agent after left anterior descending artery occlusion. Silicone pads were interposed between the transducer and the heart to simulate levels of thoracic wall attenuation. Samples of image data from perfused and nonperfused regions were collected; values using harmonic-fundamental frequency ratio peak and conventional harmonic gray scale intensity techniques were calculated. RESULTS: At each attenuation level, the harmonic-fundamental frequency ratio peak value of perfused myocardium was higher than that of nonperfused myocardium (P < .0001). The variance of these values was smaller than that of the gray scale intensity values (P < .0001), with smaller overlap between harmonic-fundamental frequency ratio peak values differentiating perfused and nonperfused regions. In the receiver operating characteristic curves, this analysis had better diagnostic performance than gray scale analysis. In the optimal cutoff value, harmonic-fundamental frequency ratio peak analysis provided 87% sensitivity and 91% specificity; gray scale analysis had 80% sensitivity and 78% specificity. CONCLUSIONS: Harmonic-fundamental frequency ratio peak analysis differentiated perfused from nonperfused myocardium under clinically relevant attenuation conditions and provided higher sensitivity and specificity for perfusion determination in attenuated myocardium than did gray scale intensity analysis.

Animals↗

Pharmacodynamic model for joint exogenous and endogenous corticosteroid suppression of lymphocyte trafficking.

The circadian pattern of the immune system correlates with that of circulating T-helper cells and inversely with cortisol concentrations. Corticosteroids, both endogenous and exogenous, cause lymphocyte dimunition in blood by retention of cells in the lymphatic circulation. A physiologic pharmacodynamic model was developed to describe changes in circulating lymphocytes as a function of both endogenous cortisol and methylprednisolone concentrations. The model was applied to T-helper and T-suppressor cell data collected from six asthmatic men during baseline, after single-dose, and after 6 days of 20 mg daily methylprednisolone. The model described all phases of the study well. Baseline circadian rhythm of lymphocytes was related to cortisol concentrations. Multiple-dosing of methylprednisolone caused apparent tolerance and decreased the sensitivity of lymphocytes to corticosteroids by 116% and markedly reduced endogenous cortisol concentrations. A 60% increase in circulating T-helper cells was observed which could be accounted for by dual changes in receptor sensitivity and endogenous cortisol.

Adrenal Cortex Hormones↗