Decentralized clinical trial design using blood microsampling technology for serum bioanalysis.

Background: Alternatives to phlebotomy in clinical trials increase options for patients and clinicians by simplifying and increasing accessibility to clinical trials. The authors investigated the technical and logistical considerations of one technology compared with phlebotomy. Methodology: Paired samples were collected from 16 donors via a second-generation serum gel microsampling device and conventional phlebotomy. Microsamples were subject to alternative sample handling conditions and were evaluated for quality, clinical testing and proteome profiling. Results: Timely centrifugation of blood serum microsamples largely preserved analyte stability. Conclusion: Centrifugation timing of serum microsamples impacts the quality of specific clinical chemistry and protein biomarkers. Microsampling devices with remote centrifugation and refrigerated shipping can decrease patient burden, expand clinical trial populations and aid clinical decisions.

A low cost, re-usable electricity-free infant warmer: evaluation of safety, effectiveness and feasibiliy.

Setting: Rural Rwandan hospitals, where thermoregulation is critical yet a challenge for pre-term, low-birth-weight (LBW) or sick newborns. Objective: To assess the safety, effectiveness, and feasibility of an inexpensive, reusable, non-electric warmer to complement kangaroo mother care (KMC). Methods: Prospective single-arm, non-randomized intervention study. Enrolled infants were hypothermic or at risk of hypothermia due to prematurity/LBW. Infants used the warmer in conjunction with KMC or as the sole source of external heat. Temperatures of the infant, warmer and air were measured for up to 6 h. Results: Overall, 33 patients used the warmer for 102 encounters: 43 hypothermic and 59 at risk of hypothermia. In 7/102 encounters (7%), the infant developed a temperature of >37.5°C (37.6°-38.2°C). For 43 hypothermic encounters and 59 at-risk encounters, hypothermia was corrected/prevented in respectively 41 (95%) and 59 (100%) instances. The warmer maintained goal temperature for the study duration in ⩾85% of uses. Two/12 warmers broke down after <10 uses. In no instances was the warmer used incorrectly. Conclusion: Our results are promising for this prototype design, and warrant testing on a wider scale.

Contexte : Des hôpitaux ruraux du Rwanda où la thermorégulation est cruciale mais complexe pour les nouveaux-nés prématurés, de faible poids de naissance (LBW) ou malades.Objectif : Evaluer la sécurité, l'efficacité et la faisabilité d'un réchauffeur peu coûteux, réutilisable et non électrique pour compléter la méthode kangourou (KMC).Méthode : Etude rétrospective d'intervention à un seul bras, non randomisée. Les nouveaux-nés enrôlés étaient en hypothermie ou à risque d'hypothermie liée à la prématurité ou au LBW. Les nouveaux-nés ont bénéficié du réchauffeur en conjonction avec la méthode KMC ou comme source unique de chaleur externe. Les températures des bébés, du réchauffeur et de l'air ont été mesurées pendant 6 h.Résultats : Ont bénéficié du réchauffeur 33 patients pour un total de 102 utilisations ; 43 étaient en hypothermie et 59 à risque d'hypothermie. Dans 7/102 utilisations (7%), le bébé a atteint une température de >37,5°C (37,6°­38,2°C). Dans 43 cas d'hypothermie et 59 cas à risque, l'hypothermie a été corrigée/prévenue dans 41 (95%) et 59 (100%) instances, respectivement. Le réchauffeur a maintenu la température souhaitée pendant la durée de l'étude dans ≥85% des utilisations. Deux réchauffeurs sur 12 ont été hors d'usage après moins de 10 utilisations. Il n'y a jamais eu d'utilisation incorrecte.Conclusion : Nos résultats sont prometteurs en ce qui concerne la conception de ce prototype et ils justifient une évaluation à plus grande échelle.

Marco de Referencia: En varios hospitales rurales de Rwanda, la termorregulación que es fundamental para los recién nacidos con bajo peso al nacer o enfermos, plantea dificultades.Objetivo: Evaluar la seguridad, la eficacia y la factibilidad de un dispositivo no eléctrico, de bajo costo y reutilizable que genera calor como complemento al método de la madre canguro (KMC).Métodos: Fue este un estudio prospectivo de intervención con un solo grupo, no aleatorizado. Se incluyeron lactantes que ya sea, estaban hipotérmicos o expuestos a la hipotermia debido a su prematuridad o el bajo peso al nacer. Con estos lactantes, se utilizó el calentador como fuente externa exclusiva de calor o en asociación con el KMC. Se midieron las temperaturas del lactante, el calentador y la temperatura ambiente durante un máximo de 6 h.Resultados: Se utilizó el dispositivo en 102 encuentros con 33 pacientes, de los cuales 43 estaban hipotérmicos y 59 estaban en riesgo de entrar en hipotermia. En siete de los 102 encuentros (7%), el lactante alcanzó una temperatura superior a 37,5°C (37,6°­38,2°C). La hipotermia se corrigió en 41 de los 43 encuentros con lactantes hipotérmicos (95%) y se evitó en 59 de las 59 ocasiones con bebés expuestos (100%). El calentador mantuvo la temperatura buscada durante todo el estudio en ≥85% de los encuentros en los cuales se utilizó. Dos de los 12 dispositivos exhibieron degradación después de menos de 10 utilizaciones. En ningún caso se utilizó el calentador de manera incorrecta.Conclusión: Los resultados obtenidos con este método prototipo son promisorios y se justifica realizar un ensayo clínico de mayor escala.

Facial Action Recognition Combining Heterogeneous Features via Multikernel Learning.

This paper presents our response to the first international challenge on facial emotion recognition and analysis. We propose to combine different types of features to automatically detect action units (AUs) in facial images. We use one multikernel support vector machine (SVM) for each AU we want to detect. The first kernel matrix is computed using local Gabor binary pattern histograms and a histogram intersection kernel. The second kernel matrix is computed from active appearance model coefficients and a radial basis function kernel. During the training step, we combine these two types of features using the recently proposed SimpleMKL algorithm. SVM outputs are then averaged to exploit temporal information in the sequence. To evaluate our system, we perform deep experimentation on several key issues: influence of features and kernel function in histogram-based SVM approaches, influence of spatially independent information versus geometric local appearance information and benefits of combining both, sensitivity to training data, and interest of temporal context adaptation. We also compare our results with those of the other participants and try to explain why our method had the best performance during the facial expression recognition and analysis challenge.

Clonal diversity in Haemophilus pleuropneumoniae.

Genetic diversity among 135 isolates of nine serotypes of Haemophilus pleuropneumoniae recovered from pigs with pleuropneumonia or other invasive diseases in 14 countries was estimated by multilocus enzyme electrophoresis, which detects allelic variation in structural genes. Thirty-two multilocus genotypes (electrophoretic types [ETs]) were distinguished on the basis of allele profiles at 15 enzyme loci, and 36 distinctive combinations of ET and serotype were identified. The recovery of isolates with identical properties in widely separated geographic regions and over a 20-year period indicated that the population structure of H. pleuropneumoniae is clonal. Isolates of the same ET generally shared the same serotype and electrophoretic pattern of the outer membrane proteins, but some ETs were represented by isolates of several different serotypes, outer membrane protein patterns, or both. On average, the genetic diversity among ETs of the same serotype was 56% of the total genetic diversity in the species. Isolates of serotype 1, which are unusually pathogenic, belong to a distinctive group of clones that are closely related to clones marked by serotype 9.

Antibody response of swine to outer membrane components of Haemophilus pleuropneumoniae during infection.

Sera from pigs infected with Haemophilus (Actinobacillus) pleuropneumoniae were tested for antibodies to outer membrane proteins (OMPs) of the organism by immunoblotting. Convalescent sera were produced in naturally born, colostrum-fed pigs and in cesarean-derived, colostrum-deprived pigs given H. pleuropneumoniae serotype 5 intranasally twice at 5-week intervals. Sera, collected at weekly intervals, were reacted with Sarkosyl-insoluble, OMP-enriched preparations of H. pleuropneumoniae which had been separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to nitrocellulose. Antibodies were detected to OMPs with an apparent molecular weight of 16,500 (16.5K OMP); to 29K, 38.5K, 43.5K, 45K, 49.5K, and 66.5K OMPs; and to several high-molecular-weight (greater than or equal to 94,000) OMPs, but not to the major 42K OMP. Antibodies to the heat-modifiable OMP (29K/43.5K) and the 38.5K OMP were detected in sera from noninfected pigs. Antibodies were also detected to two broad 54,000- and 95,000-molecular-weight bands which did not stain with Coomassie blue, stained with silver nitrate, resisted proteinase K digestion, and were eliminated by oxidation with sodium metaperiodate. This indicates that the 54,000- and 95,000-molecular-weight bands represent polysaccharide, possibly capsular or lipopolysaccharide immunogens. Adsorption of sera with cells from the homologous serotype 5 strain removed antibodies to the 45K, 49.5K, 66.5K, and greater than or equal to 94K OMPs and to the two polysaccharide bands, indicating that these antibodies were directed primarily to surface-exposed epitopes. When tested with OMP preparations from other serotype 5 strains, heterogeneity was apparent, both in the reactions with OMPs and with the polysaccharide bands. Silver staining of proteinase K-treated, whole-cell lysates from serotype 5 strains also indicated variable expression of the polysaccharide bands. Sera also reacted with OMPs from H. pleuropneumoniae serotypes 1 and 7; however, several OMPs and the lipopolysaccharide or polysaccharide determinants of these serotypes appeared to be type specific.

Outer membrane protein profiles of Haemophilus pleuropneumoniae.

Outer membrane protein profiles of Haemophilus pleuropneumoniae were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cells were disrupted by sonication, and outer membrane-enriched fractions were prepared by differential centrifugation and selective solubilization of the inner membrane with sodium N-lauroyl sarcosinate. Colony type, growth medium, time of harvest, and in vitro or in vivo passage had no appreciable effect on the protein profiles of the strains examined. Seven patterns were distinguished among the reference strains of the nine capsular serotypes. These patterns were based on the mobility of the major outer membrane proteins migrating in the 39,000- to 44,000-molecular-weight region of the gel, a 16K to 16.5K protein, and a heat-modifiable 29K protein. Strains of serotypes 1 and 9 had identical outer membrane protein profiles, as did strains of serotypes 2 and 6. The reference strains of the remaining five serotypes each had a distinct pattern. The outer membrane protein profiles of 95 field isolates belonging to serotypes 1, 5, 7, and 9 from swine in the midwestern United States were determined and compared with the reference patterns. The results indicate that the population of H. pleuropneumoniae is clonal, with three predominant clones distinguished by both serotype and outer membrane protein profile responsible for the majority of H. pleuropneumoniae disease occurring in swine in the United States.

Characterization of Haemophilus spp. isolated from healthy swine and evaluation of cross-reactivity of complement-fixing antibodies to Haemophilus pleuropneumoniae and Haemophilus taxon "minor group".

Of 30 sows from a herd believed to be free of Haemophilus pleuropneumoniae infection, 2 had complement-fixing antibodies to H. pleuropneumoniae serotype 5. Necropsy and microbiological examination of the two sows revealed no evidence of H. pleuropneumoniae infection; however, Haemophilus taxon "minor group" and a urease-negative, indole-positive Haemophilus sp. were isolated from numerous respiratory tract sites in both sows. Isolation of these Haemophilus spp. was facilitated by serially diluting specimens in two broth media. Pigs from a closed, respiratory disease-free herd were inoculated with four strains of Haemophilus taxon "minor group" to determine whether the organism induces antibodies which cross-react with H. pleuropneumoniae in the complement fixation test. Antigenic heterogeneity among the taxon "minor group" strains was apparent; however, antibodies cross-reacting between these strains and H. pleuropneumoniae serotypes 1 through 5 and 7 were not detected.

Microorganisms associated with pneumonia in slaughter weight swine.

The lungs of 334 pigs were obtained from two slaughter plants in Minnesota and examined in detail. Macroscopic and microscopic evaluation, direct fluorescence for Mycoplasma hyopneumoniae and bacterial culture were done on all of them and a subsample of 50 were selected for virus culture. Mycoplasma hyopneumoniae, Pasteurella multocida and Haemophilus spp. were detected in 24.0%, 34.1% and 27.0% of the lungs, commonly in conjunction with each other. One isolate of Haemophilus pleuropneumoniae serotype 2 was detected and this represents the first report of its presence in the United States. No virus was detected in any of the lungs. Lungs with both M. hyopneumoniae and Pasteurella multocida had the greatest amount of macroscopic pneumonia (9.8% of the lung). Lungs with M. hyopneumoniae or P. multocida alone had 4.9% and 5.2% of the lung involved with pneumonia respectively. Lungs with Haemophilus sp. Taxon "minor group" had 3.8% of the lung involved which was not significantly different from lungs with none of these organisms being detected (1.6%). There was a positive correlation between the extent of M. hyopneumoniae infection, as scored by FAT and the amount of macroscopic pneumonia present (r = 0.46; P less than 0.001). Likewise, there was a positive correlation between the estimated concentration of P. multocida present, as scored by the relative number of colonies on blood agar and the amount of macroscopic pneumonia present (r = 0.60; P less than 0.001). Microscopically, the amount of lymphoreticular proliferation, polymorphonuclear cells and alveolar macrophages were evaluated.(ABSTRACT TRUNCATED AT 250 WORDS)

Serotyping of Haemophilus pleuropneumoniae by rapid slide agglutination and indirect fluorescent antibody tests in swine.

One hundred and forty-one isolates of Haemophilus pleuropneumoniae from Iowa and Illinois swine were characterized morphologically and biochemically and serotyped by rapid slide agglutination (RSA) and indirect fluorescent antibody (IFA) tests. Hyperimmune antisera were produced in rabbits using inactivated whole-cell suspensions of the reference strains for H pleuropneumoniae serotypes 1 to 7 and strain 202, representing the taxon "minor group." Cross testing of the reference strains and reference antisera indicated the antisera to be essentially serotype-specific, although reactivity of some antisera with heterologous strains was observed. Cultures of the 141 isolates formed adherent or smooth colonies or mixtures of these colony forms. Adherent and smooth colony types were found in all serotypes identified. Microscopic and biochemical characteristics of all isolates were typical of those previously described for H pleuropneumoniae. The overall incidence of H pleuropneumoniae serotypes was serotype 5, 55.3%; serotype 1, 34.0%; serotype 7, 7.8%; and nontypeable, 2.8%. Comparing the 2 test procedures, 87.2% of the isolates could be typed by RSA, and 66.0% could be typed by IFA. Cross-reactions between serotype 4 antisera and serotype 5 and 7 isolates were common with the IFA test. The reactions with serotype 7, but not serotype 5, were eliminated by cross adsorption of serotype 4 antisera. There was good correlation between the 2 test procedures, but RSA was judged to be more specific and sensitive than IFA.