Vaccinia Virus among Domestic Dogs and Wild Coatis, Brazil, 2013-2015.

To determine their potential role as a source of human infection, we tested domestic dogs (urban) and wild coatis (wild) in Brazil for vaccinia virus. Our findings of positive neutralizing antibodies and quantitative PCR results for 35/184 dogs and 13/90 coatis highlight a potential public health risk.

An enzyme immunoassay for determining epidermal growth factor (EGF) in human serum samples using an ultramicroanalytical system.

Human epidermal growth factor is a small peptide consisting of 53 amino acid residues, which stimulates cell proliferation and is associated with several human carcinomas. A simple sandwich-type ultramicroELISA assay (UMELISA), based on the advantages of high affinity reaction between streptavidin and biotin has been developed for the measurement of EGF in human serum samples. Strips coated with a high affinity monoclonal antibody directed against EGF are used as solid phase, to ensure the specificity of the assay. The EGF assay was completed in 18 hr, with a measuring range of 39-2500 pg/mL. The intra- and inter-assay coefficients of variation were 4.4-7.3% and 0-5.1%, respectively, depending on the EGF concentrations evaluated. Percentage recovery ranged from 96-104%. Regression analysis showed a good correlation with the commercially available Human EGF Immunoassay Quantikine® ELISA kit (n = 130, r = 0.92, P < 0.01). The analytical performance characteristics of our UMELISA EGF endorse its use for the quantification of EGF in human serum samples.

Ischemic versus pharmacologic hepatic preconditioning.

BACKGROUND: Hepatic ischemia-reperfusion injury has a significant impact on liver resection and transplantation. Many strategies have been developed to reduce the effects of ischemia-reperfusion injury, including pharmacologic and ischemic preconditioning; however, studies comparing these two methods are lacking. MATERIAL AND METHODS: An experimental study was performed in a swine model. Eighteen swine were randomly assigned to three different groups: an ischemic preconditioning (IschPC) group, a pharmacologic preconditioning (PharmPC) group, and a control group. All animals underwent a 40-min liver ischemia, followed by 40 min of reperfusion. The IschPC group received a short period of ischemia (10 min) and a short period of reperfusion (15 min) before prolonged ischemia. The PharmPC group received inhaled sevoflurane for 30 min before prolonged ischemia. The control group did not receive any intervention before prolonged ischemia. Blood samples and liver tissue were obtained after ischemic and reperfusion periods. Injury was evaluated by measure of DNA damage (using COMET assay) and serum biochemical markers (transaminases, alkaline phosphatase, amylase, bilirubin, and C-reactive protein [CRP]). RESULTS: No significant difference was found in serum biochemical markers, except for the C-reactive protein level that was lower in the PharmPC group than in the control group soon after hepatic ischemia. Soon after prolonged ischemia, DNA damage index, both in blood samples and in liver tissue samples, was similar among the groups. However, an increase in DNA damage after reperfusion was higher in the control group than in the PharmPC group (P < 0.05). The increase in DNA damage in the IschPC group was half of that observed in the control, but this difference was not statistically significant. CONCLUSIONS: Our results suggest an early protective effect of PharmPC (lower levels of C-reactive protein soon after ischemia). The protective effect observed after reperfusion was higher with PharmPC than with ischemic preconditioning. The simultaneous use of both methods could potentiate protection for ischemia-reperfusion.

Carriage of Clostridium difficile in free-living South American coati (Nasua nasua) in Brazil.

The objective of this study was to isolate and characterize Clostridium difficile strains in stool samples from a wild urban mammal, a South American coati (Nasua nasua) in Brazil. Forty-six free-living N. nasua were trapped, and stool samples were collected. C. difficile was isolated from three (6.5%) sampled animals, two strains were toxigenic (A+B+CDT-, PCR ribotype 014/020 and 106) and one was non toxigenic (A-B-CDT-, PCR ribotype 053). The present work confirms that ring-tailed coati (N. nasua) could harbor C. difficile strains, including those PCR ribotypes commonly reported in C. difficile infection in humans.

Clostridium difficile and Clostridium perfringens from wild carnivore species in Brazil.

Despite some case reports, the importance of Clostridium perfringens and Clostridium difficile for wild carnivores remains unclear. Thus, the objective of this study was to identify C. perfringens and C. difficile strains in stool samples from wild carnivore species in Brazil. A total of 34 stool samples were collected and subjected to C. perfringens and C. difficile isolation. Suggestive colonies of C. perfringens were then analyzed for genes encoding the major C. perfringens toxins (alpha, beta, epsilon and iota) and the beta-2 toxin (cpb2), enterotoxin (cpe) and NetB (netb) genes. C. difficile strains were analyzed by multiplex-PCR for toxins A (tcdA) and B (tcdB) and a binary toxin gene (cdtB) and also submitted to a PCR ribotyping. Unthawed aliquots of samples positive for C. difficile isolation were subjected to the detection of A/B toxins by a cytotoxicity assay (CTA). C. perfringens was isolated from 26 samples (76.5%), all of which were genotyped as type A. The netb gene was not detected, whereas the cpb2 and cpe genes were found in nine and three C. perfringens strains, respectively. C. difficile was isolated from two (5.9%) samples. A non-toxigenic strain was recovered from a non-diarrheic maned wolf (Chrysocyon brachyurus). Conversely, a toxigenic strain was found in the sample of a diarrheic ocelot (Leopardus pardallis); an unthawed stool sample was also positive for A/B toxins by CTA, indicating a diagnosis of C. difficile-associated diarrhea in this animal. The present work suggests that wild carnivore species could carry C. difficile strains and that they could be susceptible to C. difficile infection.

Clostridium difficile-associated diarrhea in an ocelot (Leopardus pardalis).

The aim of this study is to report a case of Clostridium difficile-associated diarrhea in an ocelot (Leopardus pardalis) in the state of Mato Grosso do Sul, Brazil. The animal, a 24-month-old male, was referred to the Centro de Reabilitação de Animais Silvestres (CRAS) with a history of having been run over and tibia and fibula fractures. After a surgery to repair the fractures, the ocelot underwent antibiotic therapy with two doses of sodium cefovecin, during which he presented with diarrhea. A stool sample was positive for A/B toxins by a cytotoxicity assay, and a toxigenic strain of C. difficile was isolated. No other enteropathogens were detected. The association between the history, clinical signs and laboratory exams confirmed the diagnosis of C. difficile-associated diarrhea. The present report confirms C. difficile as a potential pathogen for wild felids and suggests that the C. difficile-associated diarrhea should be considered in diarrhea cases, especially when the clinical signs began after antimicrobial use.

Effectiveness of the mass use of deltamethrin-impregnated dog collars for preventing transmission of canine leishmaniasis by Lutzomyia spp.: A cluster randomized controlled trial.

Visceral leishmaniasis (VL) is a zoonosis that greatly impacts public health, especially in developing countries. The effectiveness of deltamethrin-impregnated dog collars for preventing transmission of canine leishmaniasis (CanL) by the sand fly Lutzomyia spp. in countries with high incidence of the disease, such as Brazil, remains unknown. A cluster randomized controlled trial was carried out to evaluate the effectiveness of these collars in preventing infection of dogs with Leishmania infantum. Two areas were selected in a municipality endemic for VL in Brazil: an experimental area (EA) and a control area (CA). In both areas, blood samples were collected from dogs for detection of CanL. Seronegative dogs from the EA received collars impregnated with deltamethrin 4%. After six months, the dogs were retested for CanL and the incidence of the disease, the relative risk, and the effectiveness of collaring determined. In the baseline survey, 1020 dogs from the EA and 589 from the CA were examined. The prevalence calculated for EA was 4.41% (CI95% 3.27-5.91), and 7.13% (CI95% 5.25-9.59) for CA. In the second evaluation, 454 dogs from the EA and 292 from the CA were examined. The person-time incidence calculated for EA was 3.51% (CI95% 1.30-7.48), and for CA was 11.81% (CI95% 7.03-18.23). The difference between the calculated incidences was statistically significant (p = 0.009). The relative risk was 0.2972 (CI95% 0.1204-0.7339; p = 0.009), and the efficacy of the proposed measure was estimated in 70.27%. This report evaluated the effectiveness of the mass use of deltamethrin-impregnated dog collars in areas of transmission of VL by Lutzomyia spp. Collaring was able to protect dogs and may represent an effective strategy that could be implemented to control infection in both dogs and humans in Brazil and perhaps other countries where Lutzomyia spp. is the main vector of the disease.

An enzyme immunoassay for determining 17alpha-hydroxyprogesterone in dried blood spots on filter paper using an ultramicroanalytical system.

BACKGROUND: 17alpha-hydroxyprogesterone has been used for the diagnosis of congenital adrenal hyperplasia (CAH) in the newborn period. METHODS: A simple and rapid competitive ultramicro ELISA assay based on competition between 17-OHP-alkaline phosphatase conjugate and 17-OHP in blood specimens for a limited number of binding sites on specific polyclonal rabbit anti-17-OHP antibodies, has been developed for the measurement of 17-OHP in dried blood spots on filter paper. The assay buffer contains danazol to displace 17-OHP from steroid-binding proteins. RESULTS: The 17-OHP assay was completed in 3 h, with a measuring range of 10-250 nmol/l. The intra- and inter-assay CV were 5.5-8.2% and 6.4-9.1%, respectively, depending on the 17-OHP concentrations. The recovery ranged from 98-103%. Of 3750 newborn samples collected on filter paper, 903 from the national neonatal screening program were analyzed, and the mean 17-OHP concentration was 12.2 nmol/l. Our assay showed high Pearson and concordance correlations with the commercially available ICN Neoscreen ELISA 17alpha-hydroxyprogesterone kit. CONCLUSIONS: The analytical performance characteristics of our 17-OHP Neonatal UMELISA suggest that it can be used for the neonatal screening of CAH.

Retroperitoneal Ectopic Pregnancy.

BACKGROUND: Retroperitoneal ectopic pregnancies are rare and have a tendency to implant along major vessels of the abdomen and pelvis. Clinical manifestations can vary from no symptoms to hypovolemic shock and death. CASE: A 30-year-old woman presented in hypovolemic shock with left flank and abdominal pain after attempting to medically terminate her first-trimester pregnancy. Intraoperative findings revealed normal pelvic organs with a massive retroperitoneal hematoma containing chorionic villi confirmed on histopathology. CONCLUSIONS: In a woman with a pregnancy of unknown location presenting with worsening flank pain, a retroperitoneal ectopic pregnancy must be considered. Prompt diagnosis and management in a low-resource setting presents additional challenges.

Programa cubano de pesquisa neonatal de la hiperplasia suprarrenal congénita: una realidad. 2005-2014 / Cuban neonatal screening program for congenital adrenal hyperplasia: a reality. 2005­2014

Introducción: la hiperplasia suprarrenal congénita (HSC) es el desorden adrenal más común en la infancia y la causa más frecuente de ambigüedad sexual. La forma clásica, que representa los casos más severos de este déficit, se asocia en un 75 % con pérdida de sal. Por otra parte, en los recién nacidos (RN) del sexo femenino se pueden presentar grados severos de virilización de genitales. Objetivo: presentar los resultados (durante diez años), del Programa Cubano de Pesquisa Neonatal (PN) de la HSC, soportado en un procedimiento inmunoenzimático desarrollado en Cuba. Resultados: en el período de enero 2005 a diciembre 2014, se han estudiado 1 140 882 RN y se detectaron 56 niños con HSC, para una incidencia de 1:20 373 RN. La cobertura del programa se ha incrementado hasta llegar en el año 2013 al 99.34 % de todos los RN cubanos. Conclusiones: la existencia del Programa Cubano de PN de HSC, ha permitido estimar la incidencia e incrementar el conocimiento de esta enfermedad. La PN ha posibilitado el diagnóstico precoz en la variedad perdedora de sal, contribuyendo a la disminución de la mortalidad infantil. El Programa ha favorecido a pacientes con formas virilizantes de la enfermedad, mediante la asignación correcta del sexo

Introduction: Congenital Adrenal Hyperplasia (HSC) is the most common adrenal disorder in childhood and the most frequent cause of sexual ambiguity. The classic form, which represents the most severe causes of this deficit, is associated in 75 % with loss of salt. On the other hand, in the NB of the female sex that present severe degrees of virilization of the genitals. Objective: To present result of the application for ten years of the Cuban Neonatal Research Program of the HSC, supported by an inmmunoenzymatic procedure developed in Cuba. Result: In the period from January 2005 to December 2014, using the UMELISA 17 OH PROGESTERONA NEONATAL, 1 140882 RN were detected, for an incidence of 1:20373 RN. The coverage of the program has been increasing until2013 reaching 99.34 % of all Cuban RN. Conclusion: The existence of the Cuban HSC PN Program, has allowed estimating the incidence and increase knowledge of this disease in our country. PN has made possible the early diagnosis of patients with the salt losing variety, contributing to decrease in infant mortality nationwide. The Program has favored patients with virilizing form of the disease, through the correct assignment of sex

From Cellulosic Based Liquid Crystalline Sheared Solutions to 1D and 2D Soft Materials.

Liquid crystalline cellulosic-based solutions described by distinctive properties are at the origin of different kinds of multifunctional materials with unique characteristics. These solutions can form chiral nematic phases at rest, with tuneable photonic behavior, and exhibit a complex behavior associated with the onset of a network of director field defects under shear. Techniques, such as Nuclear Magnetic Resonance (NMR), Rheology coupled with NMR (Rheo-NMR), rheology, optical methods, Magnetic Resonance Imaging (MRI), Wide Angle X-rays Scattering (WAXS), were extensively used to enlighten the liquid crystalline characteristics of these cellulosic solutions. Cellulosic films produced by shear casting and fibers by electrospinning, from these liquid crystalline solutions, have regained wider attention due to recognition of their innovative properties associated to their biocompatibility. Electrospun membranes composed by helical and spiral shape fibers allow the achievement of large surface areas, leading to the improvement of the performance of this kind of systems. The moisture response, light modulated, wettability and the capability of orienting protein and cellulose crystals, opened a wide range of new applications to the shear casted films. Characterization by NMR, X-rays, tensile tests, AFM, and optical methods allowed detailed characterization of those soft cellulosic materials. In this work, special attention will be given to recent developments, including, among others, a moisture driven cellulosic motor and electro-optical devices.

A cellulose liquid crystal motor: a steam engine of the second kind.

The salient feature of liquid crystal elastomers and networks is strong coupling between orientational order and mechanical strain. Orientational order can be changed by a wide variety of stimuli, including the presence of moisture. Changes in the orientation of constituents give rise to stresses and strains, which result in changes in sample shape. We have utilized this effect to build soft cellulose-based motor driven by humidity. The motor consists of a circular loop of cellulose film, which passes over two wheels. When humid air is present near one of the wheels on one side of the film, with drier air elsewhere, rotation of the wheels results. As the wheels rotate, the humid film dries. The motor runs so long as the difference in humidity is maintained. Our cellulose liquid crystal motor thus extracts mechanical work from a difference in humidity.

Newborn screening for congenital adrenal hyperplasia in Cuba: six years of experience.

BACKGROUND: Since 2005, a newborn screening program for congenital adrenal hyperplasia (CAH) by measuring 17-alpha-hydroxyprogesterone (17OHP) in dried blood spots was introduced in Cuba. METHODS: The hormone was measured by the 17OHP Neonatal UMELISA method, in samples collected on the 5th day as average. Confirmatory test was performed to those neonates with 17OHP values above 55 nmol/l. Some perinatal factors that can influence on 17OHP levels were studied. RESULTS: From January 2005 to December 2010, 621,303 newborns were screened and 39 CAH cases were detected. Coverage of the program reached 98%. The incidence of CAH in Cuba was 1:15,931, similar to that reported by other programs. A recall for suspected CAH was performed in 10,799 cases (1.74%). Therapy in classical CAH patients was started at the mean age of 22 days. 17OHP levels were significantly higher in newborns with lower birth-weight (BW) and/or gestational age (GA). In addition, 17OHP values were affected by the gender, twin status or mode of delivery. CONCLUSIONS: In Cuba, the nationwide newborn screening program has allowed the early detection of CAH. The use of an optimized cut-off level for BW or GA could lead to a reduction in the percentage of recalled babies.

Diagnóstico de animais ilegais recebidos no centro de triagem de animais silvestres de Belo Horizonte, Estado de Minas Gerais, no ano de 2011 / Diagnosis of illegal animals received at the wildlife rehabilitation center of Belo Horizonte, Minas Gerais State, Brazil in 2011

O centro de triagem de animais silvestres em Belo Horizonte (CETAS-BH) é um dos órgãos responsáveis pelo recebimento de animais silvestres em Minas Gerais (MG). O conhecimento da fauna mantida ilegal é ferramenta importante para a conservação dos animais silvestres, pois permite o aprimoramento da educação ambiental e das ações fiscalizadoras. Objetivou-se diagnosticar as espécies da fauna silvestre apreendidas ou entregues voluntariamente no CETAS-BH e analisar espacialmente sua distribuição. As espécies foram identificadas e classificadas de acordo com a chave taxonômica e o risco de extinção. Na análise descritiva espacial, foram identificadas as coordenadas geográficas dos municípios dos animais para a construção dos mapas de distribuição e de densidade de Kernel. No ano de 2011, foram recebidos 7.426 animais vivos, dos quais 91,5% eram aves, 7% répteis e 1,5% mamíferos. Verificou-se ampla variedade de espécies (166), correspondendo às aves a maioria (79,5%). As espécies mais recebidas foram Sicalis flaveola e Saltator similis. Do total de espécies recebidas, 15% estavam ameaçadas de extinção. A principal procedência foi apreensão (82,7%) e, delas, 79% foram realizadas pela Polícia Militar do Meio Ambiente. Os animais eram de 94 municípios de MG. A análise espacial identificou a Região Metropolitana de BH como o local de maior concentração de ocorrências. A partir dos resultados obtidos, espera-se o aprimoramento e a intensificação das ações de educação ambiental e de fiscalização para essas áreas específicas.

The wildlife rehabilitation center in Belo Horizonte (CETAS - BH) is one of the responsible for the receipt of wild animals in Minas Gerais (MG). The knowledge about the animals traffic is an important instrument for the wild species conservation as it allows the improvement of environmental education and the enforcement actions. The research aimed to diagnose the species of wildlife seized or voluntarily surrendered in CETAS-BH and analyze their spatial distribution. The species were identified and classified according to the taxonomic key and the risk of extinction. Descriptive spatial analysis identified the geographic coordinates of the cities where the animals were found to make maps showing their spatial distribution and the Kernel density. In 2011 were received 7426 live animals, 91.5 % of these birds, 7% reptiles and 1.5% mammals. It was found a range of species (166), most of them birds (79.5%). The most received species were Sicalis flaveola and Saltator similis. Fifteen percent of the species were endangered. The main category of receiving was the seizure (82.7%), and 79% of them were conducted by Environment Police. The animals were from 94 cities in the state of Minas Gerais. The spatial analysis identified the Belo Horizonte Metropolitan Region as the region with the highest concentration of occurrences. In this context, this study suggests the improvement and intensification of environmental education and supervision in those specific areas.

Emprego do saco herniário no reforço parietal nas hérnias inguinais indiretas do adulto / Repair of inguinal indirect hernia using the hernial sac in reinforcement wall of adult

O estudo prospectivo de oitenta pacientes com hérnia inguinal indireta primária teve como objetivo determinar as possíveis vantagens e desvantagens do uso do saco herniário próprio como um reforço adicional da fáscia transversal e do anel interno, no reparo cirúrgico da hérnia inguinal indireta, num período de acompanhamento de 24 meses após a operação. Todos os pacientes eram do sexo masculino, na faixa etária de 18 a 65 anos, com o diâmetro do anel inguinal variando de 1,0cm a 3,5cm, correspondente ao tipo 2 da classificação de Nyhus para hérnias ínguino-femorais. Foram divididos em dois grupos de quarenta cada e operados pelo mesmo cirurgião, de março de 1995. No grupo A, o reforço parietal posterior foi feito pela sutura do arco do transverso ao ligamento iliopúbico. No grupo B, acrescentou-se o saco herniário próprio, aberto e livre, fixado sobre o citado reforço às estruturas músculo-aponeuróticas do triângulo inguinal. A preparação e sutura do saco herniário foi de fácil execução, acrescentando um tempo médio de nove minutos à operação. Na comparação entre os grupos A e B, os testes estatísticos demonstraram não haver diferença significativa entre as médias de idade, peso, altura e de diâmetro do anel inguinal interno. As complicações pós-operatórias foram semelhantes nos dois grupos, sendo estas de fácil resolução. A diferença na recidiva precoce de dois casos (5 por cento) no grupo A para um caso (2,5 por cento) no grupo B não é estatisticamente significativa. No exame histopatológico dos sacos herniários, verificou-se que suas paredes constituem-se de tecido conjuntivo, vasos sanguíneos, células adiposas e, em 25 por cento dos casos, de fibras musculares lisas. Na reoperação do paciente número 12, do grupo B, observou-se que o saco herniário suturado, há 19 meses na região inguinal, apresentava-se como uma camada espessa e resistente, firmemente aderida à fáscia transversal. A proposta de utilização do saco herniário, como um reforço da hérnia inguinal, mostrou-se um recurso de fácil e rápida execução, com custo operacional baixo, não expondo o paciente ao aumento do número de complicações

Uso do saco herniário autólogo no erforço das hernioplastias incisional e inguinal / Use of autologous hernia sac in the reinforcment of inguinal and incisional hernia plastic surgery

A carência de recursos protéticos em nosso mei .o, sua difícil aquisição e os maiores riscos no seu uso nos levaram ao emprego de uma plástica na correção da hérnia incisíonal longitudinal e que no evoluir das idéias e das experiências foi também estendido à hemiorrafia inguínal Trata-se do aproveitamento do saco hemiárío no reforço paríetal das hérnias incísional e ínguinal como uma prótese autóloga, inócua e que não demonstrou nenhum sinal de aumento de complicações. Analisamos os prontuários de 132 pacientes (31 homens e 1O1 mulheres) operados de hérnia incisional pela técnica da transposição peritônio-aponeurótíca bilateral (Transpaib) e que foram acompanhados no pós-operatório tardio. Todas as hérnias eram longitudinais e apresentavam volumes médios e grandes, predominando em pacientes obesos e com perda de domicílio. O tempo de controle variou de 6 meses a 22 anos, com média de quatro anos e dez meses. Os tempos opera- tórios foram padronizados, sem o uso prévio de pneumoperitônio. Houve uma taxa de recidiva média de 13,22 por cento e todos os pacientes apresentaram hérnias recidivadas peque- nas e nos ângulos da cicatriz bem menores do que as originais. Comparando os nossos resultados com a literatura, a técnica em estudo tem o menor índice de recidiva. Ela oferece uma excelente alternativa aos cirurgiões, é exeqüível, sem auxílios tecnológicos, pode ser realizada em qualquer ambiente modesto, mas responsável e por cirurgiões de experiência mediana. No tocante aos conhecimentos aplicados à hérnia inguinal, avan- çamos no estudo histopatológico e observamos que o saco hemiário é constituído de peritônio, tecido fibroso, colágeno, parte de aponeurose, fibras reticulares, elásticas e de músculo liso. Por isso tudo parece-nos que o melhor reforço é obtido à custa dos tecidos do próprio saco hemiário.

Emprego do saco herniário, no reforço parietal nas hérnias inguinais indiretas do adulto / Hernioplasty using the hernial sac in repair of inguinal hernia in adult

O estudo prospectivo de 80 pacientes operados teve como objetivo determinar as possíveis vantagens e desvantagens do uso do saco herniário próprio como um reforço adicional da fáscia transversal e do anel interno, no reparo cirúrgico da hérnia inguinal indireta. Os pacientes foram divididos em dois grupos de 40 cada. No grupo A, o reforço parietal posterior foi feito pela sutura do arco do transverso ao ligamento iliopúbico. No grupo B, acrescentou-se o saco herniário próprio, aberto e livre, fixado sobre o citado refoço às estruturas músculo-aponeuróticas do triângulo inguinal. A preparaçäo e sutura do saco herniário foi de fácil execuçäo, acrescentou um tempo médio de nove minutos à operaçäo. Näo houve aumento do número de complicaçöes no grupo B. No exame histopatológico dos sacos herniários, verificou-se que suas paredes constituem-se de tecido conjuntivo, vasos sanguíneos, células adiposas e, em 25 por cento dos casos, também de fibras musculares lisas. Na reoperaçäo do paciente número 12, do grupo B, verificou-se que o saco herniário suturado, há 19 meses na regiäo inguinal, apresentava-se como uma camada espessa e resistente, firmemente aderida à fáscia transversal. A proposta de utilizaçäo do saco herniário, como um reforço da hérnia inguinal, mostrou-se como um recurso de fácil e rápida execuçäo, com custo operacional baixo, näo expondo o paciente ao aumento do número de complicações.