Cryopreservation of Coffea canephora Pierre ex A. Froehner seeds: importance of drying rate and moisture content.

Seeds of the Coffea canephora species are considered more recalcitrant than those of the Coffea arabica species. They tolerate partial dehydration and they cannot be stored in conventional seed banks at -18ºC because they are sensitive to freezing temperatures. Cryopreservation is a reliable method for storing seeds with these characteristics for long periods. However, preliminary studies are necessary to determine ideal storage conditions. The aim of this study was to investigate the ideal physical and physiological conditions for cryopreservation of Coffea canephora seeds to reduce seed mortality caused by the formation of intracellular ice crystals and to avoid cell damage caused by excessive desiccation. Seeds were subjected to rapid drying in silica gel and slow drying in saturated NaCl solution to moisture contents of 0.20, 0.25, and 0.28 g.g-¹ (dry basis), followed by direct immersion in liquid nitrogen for rapid freezing. Physiological and biochemical analyses were performed to evaluate seed quality before and after cryopreservation. Rapid drying of Coffea canephora seeds to values near 0.20 g.g-1 (db) does not cause reduction in physiological quality. Moisture content of 0.25 g.g-¹ brings about greater survival of Coffea canephora seeds after cryopreservation. Drying rate affects the physiological quality of cryopreserved Coffea canephora seeds, and rapid drying in silica gel is more favorable than slow drying in a saturated NaCl solution. The activity of the enzymes catalase, esterase, glutamic oxaloacetic transaminase, and polyphenol oxidase are indicators of seed quality for Coffea canephora seeds subjected to cryopreservation

Tinea Capitis in Adults. A Diagnosis to Consider?

A tinea capitis, uma infecção por fungos dermatófitos do couro cabeludo, é uma patologia rara no adulto, mesmo em áreas geográficas onde a doença é prevalente. Nestes casos está muitas vezes, mas não exclusivamente, associada a imunodepressão e habitualmente surge por exposição ao agente infeccioso em contactantes próximos. São fundamentais uma história clínica e um exame objectivo minuciosos que permitam equacionar a tinha do couro cabeludo como hipótese diagnóstica e proceder à colheita de amostras para exame micológico. Esta metodologia pode evitar a realização de exames complementares exaustivos ou a prescrição de tratamentos inadequados. Os autores descrevem três casos de tinea capitis em mulheres adultas – dois casos de infecção por Microsporum audouinii e um caso por Trichophyton soudanense. É apresentado o registo iconográfico dos casos e é efectuada breve revisão da literatura

Diagnostic accuracy of single baseline measurement of Elecsys Troponin T high-sensitive assay for diagnosis of acute myocardial infarction in emergency department: systematic review and meta-analysis

Published onlineJournal ArticleMeta-AnalysisResearch Support, Non-U.S. Gov'tReviewOBJECTIVE: To obtain summary estimates of the accuracy of a single baseline measurement of the Elecsys Troponin T high-sensitive assay (Roche Diagnostics) for the diagnosis of acute myocardial infarction in patients presenting to the emergency department. DESIGN: Systematic review and meta-analysis of diagnostic test accuracy studies. DATA SOURCES: Medline, Embase, and other relevant electronic databases were searched for papers published between January 2006 and December 2013. STUDY SELECTION: Studies were included if they evaluated the diagnostic accuracy of a single baseline measurement of Elecsys Troponin T high-sensitive assay for the diagnosis of acute myocardial infarction in patients presenting to the emergency department with suspected acute coronary syndrome. STUDY APPRAISAL AND DATA SYNTHESIS: The first author screened all titles and abstracts identified through the searches and selected all potentially relevant papers. The screening of the full texts, the data extraction, and the methodological quality assessment, using the adapted QUADAS-2 tool, were conducted independently by two reviewers with disagreements being resolved through discussion or arbitration. If appropriate, meta-analysis was conducted using the hierarchical bivariate model. RESULTS: Twenty three studies reported the performance of the evaluated assay at presentation. The results for 14 ng/L and 3-5 ng/L cut-off values were pooled separately. At 14 ng/L (20 papers), the summary sensitivity was 89.5% (95% confidence interval 86.3% to 92.1%) and the summary specificity was 77.1% (68.7% to 83.7%). At 3-5 ng/L (six papers), the summary sensitivity was 97.4% (94.9% to 98.7%) and the summary specificity was 42.4% (31.2% to 54.5%). This means that if 21 of 100 consecutive patients have the target condition (21%, the median prevalence across the studies), 2 (95% confidence interval 2 to 3) of 21 patients with acute myocardial infarction will be missed (false negatives) if 14 ng/L is used as a cut-off value and 18 (13 to 25) of 79 patients without acute myocardial infarction will test positive (false positives). If the 3-5 ng/L cut-off value is used, <1 (0 to 1) patient with acute myocardial infarction will be missed and 46 (36 to 54) patients without acute myocardial infarction will test positive. CONCLUSIONS: The results indicate that a single baseline measurement of the Elecsys Troponin T high-sensitive assay could be used to rule out acute myocardial infarction if lower cut-off values such as 3 ng/L or 5 ng/L are used. However, this method should be part of a comprehensive triage strategy and may not be appropriate for patients who present less than three hours after symptom onset. Care must also be exercised because of the higher imprecision of the evaluated assay and the greater effect of lot-to-lot reagent variation at low troponin concentrations. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration number CRD42013003926.This research was funded by the South West Academic Health Science Network (AHSN) and the National Institute for Health Research (NIHR) Collaboration for Leadership for Applied Health Research and Care for the South West Peninsula

Construção e caracterização eletroquímica de imunossensor para diagnóstico de doenças em bovinos.

O objetivo desse estudo é desenvolver e investigar o comportamento de um dispositivo eletroquímico altamente sensível e reprodutível, um biossensor, capaz de realizar detecção direta de proteína, enzima ou outras biomoléculas em solução

Analyzing power in nucleon-deuteron scattering and three-nucleon forces

Three-nucleon forces have been considered to be one possibility to resolve the well known discrepancy between experimental values and theoretical calculations of the nucleon analyzing power in low energy nucleon-deuteron scattering. In this paper, we investigate possible effects of two-pion exchange three-nucleon forces on the analyzing power and the differential cross section. We found that the reason for different effects on the analyzing power by different three-nucleon forces found in previous calculations is related to the existence of the contact term. Effects of some variations of two-pion exchange three-nucleon forces are investigated. Also, an expression for the measure of the nucleon analyzing power with quartet P-wave phase shifts is presented.Comment: 11 pages including 2 eps figures, use epsfig.sty, to appear in Phys. Rev.

Neutron-3H and Proton-3He Zero Energy Scattering

The Kohn variational principle and the (correlated) Hyperspherical Harmonics technique are applied to study the n-3H and p-3He scattering at zero energy. Predictions for the singlet and triplet scattering lengths are obtained for non-relativistic nuclear Hamiltonians including two- and three-body potentials. The calculated n-3H total cross section agrees well with the measured value, while some small discrepancy is found for the coherent scattering length. For the p-3He channel, the calculated scattering lengths are in reasonable agreement with the values extrapolated from the measurements made above 1 MeV.Comment: 13 pages, REVTEX, 1 figur

Household transmission of invasive group A Streptococcus infections in England: a population-based study, 2009, 2011 to 13

Invasive group A s treptococcal infection has a 15% case fatality rate and a risk of secondary transmission. This retrospective study us ed two national data sources from England ; enhanced surveillance ( 2009) and a case management system ( 2011 - 13) to identify clusters of sever e group A streptococcal disease . 2 4 household pairs were identified. The median onset interval between cases was 2 days (range 0 - 28) with simultaneous onset in 8 pairs . The attack rate during the 30 days after first exposure to a primary case was 4 52 0 per 100000 person - years at risk (95% CI 2 900 - 673 0 ) a 19 40 ( 12 40 - 28 80 ) fold elevation over the background incidence . The theoretical number needed to treat ( NNT ) to prevent one secondary case using antibiotic prophylaxis was 2 71 ( 194 - 454 ) overall, 50 for mother - neonate pairs ( 2 7 - 3 93 ) and 8 2 for couples aged 75 years and over ( 46 - 417 ). Whilst a dramatic increased risk of infection was noted in all household contacts, increased ris k was greatest for mother - neonate pairs and couples aged 75 and over , suggesting targeted prophylaxis c ould be considered. Offering prophylaxis is challenging due to the short time interval between cases emphasising the importance of immediate notification and assessment of contacts

Differential protection induced by immunization with variable doses of a Leishmania antigenic extract against Leishmania amazonensis.

Experimental vaccines have been developed to protect against leishmaniasis using the BALB/c mice model immunized with different immunogens, but an effective vaccine still does not exist. To determine factors inherent to immunogens that might abrogate vaccine-induced efficacy, our research sought to investigate the impact of immunization using variable doses (low, medium and high) of a known soluble Leishmania antigenic extracts (SLA), associated or not with alum, in order to determine the best dose of this vaccine immunogen able to induce the best level of protection in BALB/c mice against L. amazonensis infection. This work shows that the immunization´ model using a high inoculum (100 µg) of SLA results in the best level of protection against challenge. These mice presented significant reductions in the footpad swelling and parasite load; high levels of IFN-&#947; and IL-12, and low levels of IL-4, IL-10, TGF-&#946; and Leishmania-specific IgG and IgE antibodies. Mice immunized with 50 µg of SLA present intermediate results of protection; on the other hand, mice immunized with 1 µg showed the worst results. Considering all the elements, it could be concluded that the model employing a high dose of SLA in BALB/c mice can bring about the development of a protective immune response in the animals, thus allowing for the protection against the disease. In addition, we understand that the definition of an ideal dose for each vaccine candidate appears to be fundamental to determining the phenotype of resistance and/or susceptibility in murine models to study leishmaniasis

Genetic diversity among Colletotrichum sublineolum pathotypes isolated from sorghum (Sorghum bicolor).

Sorghum anthracnose, caused by Colletotrichum sublineolum is a serious disease in Brazil. The fungus is highly variable pathogenically with many physiological races. The present study was made to evaluate the usefulness of molecular methods (SDS-PAGE, RAPD, ARDRA and rDNA sequencing) for identifying C. sublineolum pathotypes isolated from sorghum. Although protein profile (SDS-PAGE) showed a relative low level of polymorphism, differences could be observed due to presence/absence of some polypeptides as well as to intensity of some bands among pathotypes. DNA profile of arbitrarily amplified sequences (RAPD) using sixteen random primers revealed a high degree of polymorphism while amplified rDNA (ITS region and 18S gene) digested with six different restriction enzymes (ARDRA) showed similar patterns. Nucleotide sequencing analysis of the complete ITS region and partial sequencing of 18S rRNA gene were useful for revealing genetic differences among the five pathotypes. In the present study, RAPD and rDNA sequencing were the most profitable methods for identifying C. sublineolum pathotypes. A antracnose do sorgo, causada por Colletotrichum sublineolum, é séria doença, no Brasil. O fungo é altamente variável do ponto de vista patogênico e várias raças têm sido noticiadas. Este estudo foi realizado com o objetivo de avaliar a utilidade de métodos moleculares (SDS-PAGE, RAPD, ARDRA e seqüenciamento de rDNA) para identificar patótipos de C. sublineolum isolados do sorgo. Embora o perfil protéico (SDS-PAGE) tenha mostrado baixo grau de polimorfismo, foi possível observar diferenças devido à presença/ausência de alguns polipeptídios, bem como diferenças quanto à intensidade de algumas bandas. O perfil de DNA amplificado arbitrariamente (RAPD), usando dezeseis primers aleatórios, revelou elevado grau de polimorfismo entre os patótipos, enquanto que o produto da amplificação de rDNA (região ITS e gene 18S), digerido com seis enzimas de restrição diferentes (ARDRA), apresentou padrão similar. Análise de nucleotídeos da seqüência completa da região ITS e do seqüenciamento parcial do gene 18S rRNA revelaram diferenças genéticas entre os cinco patótipos. Entre os métodos testados no presente estudo, RAPD e seqüenciamento de rDNA foram mais efetivos para identificar patótipos de C. sublineolum