Profile of emergency department overuse in hospitalized patients with pulmonary disease and its impact on mortality.

INTRODUCTION AND OBJECTIVES: Portugal is one of the countries with the highest number of visits to the emergency department (ED), 31% classified as "non-urgent" or "avoidable." The objectives of our study were to evaluate the size and characteristics of patients with pulmonary disease who overuse the ED, and identify factors associated with mortality. MATERIALS AND METHODS: A retrospective cohort study was conducted, based on the medical records of ED frequent users (ED-FU) with pulmonary disease who attended a university hospital center in the northern inner city of Lisbon from January 1 to December 31, 2019. To evaluate mortality, a follow-up until December 31, 2020 was performed. RESULTS: Over 5,567 (4.3%) patients were identified as ED-FU and 174 (0.14%) had pulmonary disease as the main clinical condition, accounting for 1,030 ED visits. 77.2% of ED visits were categorized as "urgent/very urgent." A high mean age (67.8 years), male gender, social and economic vulnerability, high burden of chronic disease and comorbidities, with a high degree of dependency, characterized the profile of these patients. A high proportion (33.9%) of patients did not have a family physician assigned and this was the most important factor associated with mortality (p<0.001; OR: 24.394; CI 95%: 6.777-87.805). Advanced cancer disease and autonomy deficit were other clinical factors that most determined the prognosis. CONCLUSIONS: Pulmonary ED-FU are a small group of ED-FU who constitute an aged and heterogeneous group with a high burden of chronic disease and disability. The lack of an assigned family physician was the most important factor associated with mortality, as well as advanced cancer disease and autonomy deficit.

Transfusion transmissible pathogens are prevalent in healthy cats eligible to become blood donors.

OBJECTIVES: This study aims to determine the prevalence of subclinical infectious agents considered core pathogens for worldwide screening in healthy, client-owned, indoor cats eligible to become blood donors in Spain and Portugal. MATERIALS AND METHODS: Blood samples of healthy, indoor, domestic cats selected to be potential blood donors were tested for feline leukaemia virus antigens, feline immunodeficiency virus antibodies and polymerase chain reactions for Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum, Candidatus Mycoplasma turicensis, feline leukaemia virus provirus, Leishmania spp. and Bartonella spp. Not all donors were tested for all agents. RESULTS: Overall, 5105 healthy indoor cats were tested and 8.1% (414/5105) had at least one subclinical infectious agent that is transmissible through blood product transfusion. 1.5% (77/5105) were positive for feline leukaemia virus antigens and 2.9% (148/5105) were positive for feline immunodeficiency virus antibodies, therefore they were excluded as donors. The overall prevalence of haemoplasmas in feline leukaemia virus and feline immunodeficiency virus SNAP-negative feline blood donors was 3.7% (181/4880) [1.3% (63/4880) were positive for Mycoplasma haemofelis; 2.3% (112/4880) to Candidatus Mycoplasma haemominutum and 0.12% (6/4880) to Candidatus Mycoplasma turicensis]. The prevalence of feline leukaemia provirus was 5.2% (9/173) and of Bartonella spp. was 0.2% (2/1122). None of the 173 cats were positive for Leishmania spp. CLINICAL SIGNIFICANCE: The prevalence of many transfusion transmissible pathogens was relatively high in this healthy, client-owned, indoor cats eligible to become blood donors. Performing an extended screening panel that includes at least polymerase chain reactions for Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum, Candidatus Mycoplasma turicensis, feline leukaemia virus provirus, and Bartonella spp., in addition to feline leukaemia virus antigens and feline immunodeficiency virus antibodies, is recommended in pet blood banks from analogous regions.

Productive characteristics in dairy buffalo (Bubalus bubalis) in the Eastern Amazon / Características produtivas em búfalos leiteiros (Bubalus bubalis) na Amazônia

The objective of this study was to determine factors affecting phenotypic characteristics related to milk production and reproductive efficiency of dairy buffalo. Phenotypic records of 2,459 Murrah, Mediterranean and crossbred buffaloes from the Embrapa Eastern Amazônia herd, from 1969 to 2013 were used. Traits included: total milk production during lactation (TMP), milk production corrected for 305 days (P305); cow weight at calving (CW), fat milk percentage (F), age at first calving (AFC), calving interval (CI), calf birth weight (BW), as well as lactation (LP) and service periods (SP). Most traits were affected by genetic group as well as year and season or interaction between them. Genetic group affected F, CI, SP and CW. Very young and older cows showed poorer production traits. There was interaction between season and year for F, CI and CW. Correlations traits were generally medium to low. The principal component analysis showed that higher fat milk percentage was associated with higher percentage of Murrah breed, longer lactation period and older cows. Phenotypic characteristics in buffaloes reared in the Brazilian Amazon were significantly affected by genetic and management factors. This information can be used to help build sustainable production systems for the region and guide farmer decisions.(AU)

O objetivo deste estudo foi determinar os fatores que afetam as características fenotípicas relacionadas à produção de leite e a eficiência reprodutiva de búfalas leiteiras. Foram utilizados os registros fenotípicos de 2.459 búfalas das raças Murrah, Mediterrâneo e seus cruzamentos do rebanho da Embrapa Amazônia Oriental, de 1969 a 2013. As características incluídas foram: produção total de leite durante a lactação (PTL), produção de leite corrigida para 305 dias (P305), peso da búfala ao parto (PV), percentual de gordura do leite (G), idade ao primeiro parto (IPP), intervalo entre partos (IP), peso do bezerro ao nascimento (PN), assim como os períodos de lactação (PL) e de serviço (PS). A maioria das características foi afetada pelo grupo genético, bem como pelo ano e pela estação ou pela interação entre eles. O grupo genético afetou o G, IP, PS e PV. Búfalas muito jovens e mais velhas mostraram características de produção mais baixas. Houve interação entre estação e o ano para G, IP e PV. As correlações entre as características foram geralmente de médias a baixas. A análise de componentes principais mostrou que a maior porcentagem de gordura no leite foi associada com a maior porcentagem de sangue Murrah, com o maior período de lactação e com búfalas mais velhas. As características fenotípicas das búfalas criadas na Amazônia brasileira foram significativamente afetadas por fatores genéticos e de manejo. Essas informações podem ser usadas para ajudar a construir sistemas de produção sustentáveis para a região e orientar as decisões dos produtores.(AU)

Ménétrier's disease : a case of successful treatment using long-acting octreotide.

The authors describe a 31 years old male, admitted for hematemesis, epigastric pain and lower limb edema. Laboratorial data showed haemoglobin 18.4g/dl, total proteins 2.8g/dl, albumin 1.6g/dl and hipogammaglobulinaemia. 24h urinary proteins were normal. HIV and CMV serology were negative. Upper GI endoscopy revealed markedly enlarged gastric folds covered by abundant exudative fluid. Endoscopic ultrasound showed ascites, pleural effusion and gastric wall thickening with mucosa expansion and intact submucosa. In gastric biopsies foveolar hyperplastic and regenerative mucosa were observed being suggestive of Ménétrier´s disease. Helicobacter pylori was not detected. Albumin replacement and diuretics corrected anasarca and long-acting octreotide was instituted. Nine months later, the patient was asymptomatic, serum proteins were normal (albumin 4.6g/dl and total proteins 6.5g/dl), signs of endoscopic improvement were observed with marked reduction in gastric folds and mucosal inflammation and no ultrastructural changes were detected in gastric specimens sent for electron microscopy. Ménétrier´s Disease (MD) is a rare form of hypertrophic gastropathy characterized by massive enlargement of gastric folds causing marked protein exudation. The increase in tight junction diameter is the most consistent ultraestrutural change. Octreotide is a somatostatin analogue that acts by modulating TGFαEGFR pathway, which has been associated with the pathogenic mechanisms. As well as other cases reported in literature, this case report highlights the potential of long-acting octreotide for MD treatment avoiding more expensive therapies like cetuximab and gastrectomy.

Genetic evaluation of milking buffaloes (Bubalus bubalis): bull ranking / Avaliação genética de búfalas leiteiras (Bubalus bubalis): classificação de touros

The objective of this study was to evaluate genetic aspects related to production and reproductive efficiency of Murrah and Mediterranean buffaloes and their crosses. A ranking of bulls from Embrapa Eastern Amazonia was also composed to guide assisted mating. Birth records of 2,322 Murrah, Mediterranean, and crossbred buffaloes from the Embrapa Eastern Amazon herd, from 1953 to 2013, as well as information on production and reproductive traits were used. Genetic analyzes were performed in the WOMBAT software using the animal model with two-trait analysis. While heritability (h2) for total milk production (TMP) and fat milk percentage (F) were generally high, for reproductive traits h2 tended to be low. Genetic correlations for TMP with the other traits were low and negative, except for TMP with calving interval (CI) and service period (SP) in the Mediterranean breed and with age at first calving (AFC) and SP in crossbred, which were positive and high. Bull 1001 had high predicted transmitting ability (PTA) for TMP, so it should transmit a greater volume of milk to his offspring, although it had a lower PTA for F. There was sufficient variability within the herd to work with genetic management for both production and reproductive efficiency.(AU)

O objetivo deste estudo foi avaliar os aspectos genéticos relacionados à produção e à eficiência reprodutiva de búfalas das raças Murrah, Mediterrâneo e suas cruzas. Uma classificação de touros da Embrapa Amazônia Oriental também foi composta para orientar os cruzamentos assistidos. Foram utilizados 2.322 registros de nascimento de búfalas das raças Murrah, Mediterrâneo e cruzadas do rebanho da Embrapa Amazônia Oriental, de 1953 a 2013, bem como características produtivas e reprodutivas. As análises genéticas foram realizadas pelo software Wombat, utilizando-se o modelo animal com análise de duas características. Enquanto a herdabilidade (h 2 ) para a produção total de leite (PTL) e para a porcentagem de gordura (G) foi alta, para as características reprodutivas a h 2 tendeu a ser baixa. As correlações genéticas da PTL com as demais características foram baixas e negativas, exceto para a PTL com intervalo entre partos (IP) e período de serviço (PS) na raça Mediterrâneo e com idade ao primeiro parto (IPP) e PS nas cruzadas, que foram positivas e altas. O touro 1001 apresentou alta capacidade de transmissão predita (CTP) para a PTL, então deve transmitir um maior volume de leite para seus descendentes, embora com um menor conteúdo transmissível de CTP para G. Portanto, existe variabilidade suficiente dentro do rebanho para trabalhar com o manejo genético para a produção e a eficiência reprodutiva.(AU)

Ocorrência de anticorpos anti-Leishmania spp. em felinos em área endêmica do estado de São Paulo / Occurrence of anti-Leishmania spp antibodies in felines in an endemic area of the State of São Paulo

A leishmaniose visceral (LV) é uma zoonose de grande impacto em saúde pública. A infecção nos gatos tem sido relatada nos países onde a doença é endêmica. Seu papel como reservatório não está satisfatoriamente elucidado, embora a transmissão do parasito de um felino infectado para vetor tenha sido reportada por xenodiagnóstico. O objetivo do trabalho foi avaliar a presença de anticorpos anti-Leishmania spp. em animais da espécie felina em área endêmica para LV (Bauru-SP), por meio dos testes sorológicos de reação de imunofluorescência indireta (RIFI) e ensaio imunoenzimático (ELISA), e associá-los às variáveis: gênero, idade, raça e forma de criação. Foram testados soros de 276 felinos, dos quais 82 foram reagentes pelo método ELISA (29,71%), 17 pelo RIFI (6,15%) e 10 em ambos os testes (3,6%). Houve associação estatística significativa para a variável forma de criação, em que 100% dos animais errantes foram soropositivos a pelo menos um dos testes (P<0,005). Tal associação não foi encontrada para as demais variáveis analisadas (P>0,05). Não houve concordância entre o resultado dos testes, pois o método ELISA é mais sensível que o método RIFI.(AU)

Visceral leishmaniasis (VL) is a zoonosis with a great impact on public health. Infection in cats has been reported in countries where the disease is endemic. Its role as reservoir is not satisfactorily elucidated, although transmission of the parasite from an infected feline to vector has been reported by xenodiagnosis. The objective of this study was to evaluate the presence of anti-Leishmania spp antibodies in feline animals in an area endemic to LV (Bauru-SP), using the serological tests of Indirect Immunofluorescence Reaction (IFR) and ELISA and variables: gender, age, race and form of creation. Samples of 276 felines were tested, of which 82 were ELISA reagents (29,71%), 17 by IFR (6,15%) and 10 in both tests (3,6%). There was a significant statistical association for the variable form of breeding, where 100% of the wandering animals were seropositive to at least one of the tests (P <0,005). Such association was not found for the other variables analyzed (P >0,05). There was no concordance between the results of the tests, since the ELISA method is more sensitive than the RIFI method.(AU)

Incidence, characteristics and geographical distributions of canine and human non-Hodgkin's lymphoma in the Porto region (North West Portugal).

Lymphoma is one of the most common neoplasms in dogs and it is one of the top five causes of cancer-related deaths, similar to human lymphoma. Companion animal epidemiological studies define dogs as sentinels of potential risk factors for human health, mainly due to shared environments, shorter disease latencies, and spontaneous disease. The aims of this study were to describe human and canine epidemiologic features of non-Hodgkin's lymphoma (NHL) and their similarities, and to investigate a possible geographical association in the incidence risks in the Greater Porto area, in north-western Portugal. The postal codes of human NHL patients diagnosed between 2005 and 2010 residing in the Greater Porto, Portugal, were obtained from North and Central Region Cancer Registries of Portugal. Available data from dogs diagnosed with lymphoma between 2005 and 2016 from several veterinary centres were also collected. Descriptive epidemiology, mapping cases, and age-standardised risks of NHL incidence (ASR) were determined for both species. The results showed a higher risk (P<0.05) of NHL in men (ASR men: 18.1 cases/100,000 inhabitants; women: 14.2 cases/100,000 inhabitants) and in male dogs (ASR males: 82 cases/100,000 dogs; females: 70 cases/100,000 dogs). The geographical distribution of human and canine ASR was well correlated (r=0.664, P<0.05), with the highest values for human and canine ASR detected in the same urban municipalities of the Greater Porto: Porto, Matosinhos and Maia. These findings suggest the existence of exposure similarities, supporting the relevance of cancer surveillance in pet animals as efficient tools to predict health hazards for humans.

Immunocytochemical study of canine lymphomas and its correlation with exposure to tobacco smoke.

AIM: Canine lymphoma is one of the most common canine neoplasms, but little is known regarding the effects of exposure to tobacco smoke on their biologic behavior. As cytology is the most frequent diagnostic method of canine lymphoma, the aims of this study were to perform an immunocytochemical study of canine lymphomas, including subtyping and cell proliferation analysis, and to establish their correlation with tobacco smoke exposure. MATERIALS AND METHODS: A total of 23 dogs diagnosed with lymphoma were subjected to careful fine-needle biopsies of enlarged lymph nodes. The smears were air-dried, fixed with cold acetone, and immunocytochemically stained using CD3, PAX5, and Ki-67. Owners were requested to complete an epidemiologic questionnaire. RESULTS: According to the updated Kiel classification, 65% were B-cell lymphomas - three low grade (LG) and 12 high grade (HG) and 35% were T-cell -two LG and six HG. Thirteen tumors presented high Ki67 indexes (>40%) (11 HG and 2 LG), two revealed moderate ones (20-40%) (1 HG and 1 LG), and three had low indexes (≤20%) (1 HG and 2 LG). Both a significant positive correlation and a significant linear-by-linear association (p=0.018) were observed between high Ki67 indexes and smoking owners (r=0.753, p=0.002) as well as with the number of smokers in the household (r=0.641, p=0.001). Moreover, the mean percentage of Ki67+ cells from the group of "smoker owners" was statically higher (p=0.011) than that from the "non-smoker owners." CONCLUSION: The results suggest that cytological diagnosis of canine lymphomas benefits from being complemented with immunocytochemical studies that include subtyping and assessment of proliferative activity, both contributing for the prognosis and therapeutic planning. Furthermore, exposure to tobacco smoke seems to be related to the biological behavior of canine lymphomas.

Evidence-based guides in tracheostomy use in critical patients

Objetivos: Proporcionar guías de traqueostomía para el paciente crítico, basadas en la evidencia científica disponible, y facilitar la identificación de áreas en las cuales se requieren mayores estudios. Métodos: Un grupo de trabajo formado con representantes de 10 países pertenecientes a la Federación Panamericana e Ibérica de Sociedades de Medicina Crítica y Terapia Intensiva y a la Latin American Critical Care Trial Investigators Network(LACCTIN) desarrollaron estas recomendaciones basadas en el sistema Grading of Recommendations Assessment, Development and Evaluation (GRADE). Resultados: El grupo identificó 23 preguntas relevantes entre las 87 preguntas planteadas inicialmente. En la búsqueda inicial de la literatura se identificaron 333 estudios, de los cuales se escogieron un total de 226. El equipo de trabajo generó un total de 19 recomendaciones: 10 positivas (1B = 3, 2C = 3, 2D = 4) y 9 negativas (1B = 8, 2C = 1). En 6 ocasiones no se pudieron establecer recomendaciones. Conclusión: La traqueostomía percutánea se asocia a menor riesgo de infecciones en comparación con la traqueostomía quirúrgica. La traqueostomía precoz solo parece reducir la duración de la ventilación mecánica pero no la incidencia de neumonía, la duración de la estancia hospitalaria o la mortalidad a largo plazo. La evidencia no apoya el uso de broncoscopia de forma rutinaria ni el uso de máscara laríngea durante el procedimiento. Finalmente, el entrenamiento adecuado previo es tanto o más importante que la técnica utilizada para disminuir las complicaciones.(AU)

OBJECTIVES: Provide evidence based guidelines for tracheostomy in critically ill adult patients and identify areas needing further research. METHODS: A task force composed of representatives of 10 member countries of the Pan-American and Iberic Federation of Societies of Critical and Intensive Therapy Medicine and of the Latin American Critical Care Trial Investigators Network developed recommendations based on the Grading of Recommendations Assessment, Development and Evaluation system. RESULTS: The group identified 23 relevant questions among 87 issues that were initially identified. In the initial search, 333 relevant publications were identified of which 226 publications were chosen. The task force generated a total of 19 recommendations: 10 positive (1B=3, 2C=3, 2D=4) and 9 negative (1B=8, 2C=1). A recommendation was not possible in six questions. CONCLUSION: Percutaneous techniques are associated with a lower risk of infections compared to surgical tracheostomy. Early tracheostomy only seems to reduce the duration of ventilator use but not the incidence of pneumonia, the length of stay, or the long-term mortality rate. The evidence does not support the use of routine bronchoscopy guidance or laryngeal masks during the procedure. Finally, proper prior training is as important or even a more significant factor in reducing complications than the technique used.(AU)

Composición química e indicadores de calidad del frijol de soya (Glycine max) integral procesado con vapor para la alimentación de aves y cerdos / Chemical composition and quality indicators of full-fat soybean steam processed for poultry and swine feeding

Este estudio se realizó para determinar la composición química e indicadores de calidad del frijol de soya (Glycine max) integral (FSI) procesado con vapor en un autoclave bajo condiciones estándares de trabajo a 2,30 bar de presión a 103°C/30 min. Se tomaron seis muestras después del procesamiento de cada uno de tres lotes de soya provenientes de Estados Unidos. Se obtuvo la media por lote y general para cada variable con el error estándar (± EE). Las medias generales de materia seca, proteína, grasa, fibra, ceniza, Ca y P (g/100 g) y energía bruta (kcal/kg) fueron 91,40±0,40; 36,82±0,18; 20,88±0,49; 6,69±0,21; 4,80±0,05; 0,30±0,06; 0,53±0,006 y 5732±44,14, respectivamente. Hubo variaciones entre lotes para materia seca, P (P<0,01), fibra, ceniza y energía bruta (P≤0,05). Se observó predominio de lisina, arginina y leucina cuyas medias generales fueron 2,26±0,01; 2,70±0,01 y 2,81±0,01, respectivamente. Los ácidos grasos predominantes fueron oleico y linoleico con medias generales (g/100g) de 21,65±0,12 y 51,07±0,13, respectivamente. Se observaron variaciones (P≤0,01) entre lotes en la acidez y en los índices de saponificación, yodo y acidez. Las medias generales para inhibidores de tripsina, solubilidad de la proteína en KOH, actividad de ureasa y lisina reactiva fueron 3,04 ± 0,36 mg/g de muestra; 80,29 ± 1,09%; 0,05 ± 0,01% y 1,87±0,06%, respectivamente. Aunque hubo variaciones entre lotes para algunos componentes e indicadores de calidad, el FSI procesado con vapor es una fuente proteica de buena calidad, con buen contenido de lisina, grasa, energía bruta y ácidos grasos esenciales para la alimentación de aves y cerdos(AU)

This study was conducted to determine the chemical composition and quality indicators of full-fat soybean (Glycine max) (FFSB) steam processed in an autoclave under standard contitions of work of 2.30 bar of pressure and 103°C/30 min. Six samples were taken after the processing of each of three soybean batches from the United States. The average was obtained by batch and overall for each variable with the standard error of the mean (± SEM). The total averages of dry matter, protein, fat, fiber, ash, Ca and P (g/100g) and gross energy (kcal/kg) were 91.40±0.40, 36.82 ± 0.18; 20.88±0.49, 6.69 ± 0.21, 4.80±0,05, 0.30±0.06, 0.53±0.006, and 5732±44.14, respectively. There were variations among batches for dry matter, P (P<0.01), fiber, ash and gross energy (P≤0.05). There was a predominance of lysine, arginine and leucine (g/100 g) with average of 2.26±0.01, 2.70±0.01 and 2.81±0.01, respectively. The predominant fatty acids (g/100 g) were oleic and linoleic with general averages of 21.65 ± 0.12 and 51.07 ± 0.13, respectively. There were variations (P≤0.01) among batches in acidity and saponification, iodine and acidity index. The average for trypsin inhibitors, protein solubility in KOH, urease activity and reactive lysine were 3.04 ± 0.36 mg/g of sample; 80.29 ± 1.09%, 0.05 ± 0.01% y 1.87±0.06%, respectively. Although there were variations among batches for some components and quaility indicators, the FFSB steam processed is a protein source of good quality with good content of lysine, fat, gross energy and essential fatty acids for feeding poultry and swine(AU)

Evidence-based guides in tracheostomy use in critical patients. / Guías basadas en la evidencia para el uso de traqueostomía en el paciente crítico.

OBJECTIVES: Provide evidence based guidelines for tracheostomy in critically ill adult patients and identify areas needing further research. METHODS: A task force composed of representatives of 10 member countries of the Pan-American and Iberic Federation of Societies of Critical and Intensive Therapy Medicine and of the Latin American Critical Care Trial Investigators Network developed recommendations based on the Grading of Recommendations Assessment, Development and Evaluation system. RESULTS: The group identified 23 relevant questions among 87 issues that were initially identified. In the initial search, 333 relevant publications were identified of which 226 publications were chosen. The task force generated a total of 19 recommendations: 10 positive (1B=3, 2C=3, 2D=4) and 9 negative (1B=8, 2C=1). A recommendation was not possible in six questions. CONCLUSION: Percutaneous techniques are associated with a lower risk of infections compared to surgical tracheostomy. Early tracheostomy only seems to reduce the duration of ventilator use but not the incidence of pneumonia, the length of stay, or the long-term mortality rate. The evidence does not support the use of routine bronchoscopy guidance or laryngeal masks during the procedure. Finally, proper prior training is as important or even a more significant factor in reducing complications than the technique used.

Accuracy of ultrasound in the diagnosis of acute cholecystitis with coexistent acute pancreatitis.

PURPOSE: Acute cholecystitis and pancreatitis are acute forms of cholecystolithiasis. The presence of acute cholecystitis can lead to important changes in therapy in the early course of acute pancreatitis. The aim of this study was to identify the accuracy of ultrasonography in diagnosing acute cholecystitis with coexistent acute pancreatitis. METHODS: Subjects were all those patients admitted to our hospital with a diagnosis of acute pancreatitis between 1998 and 2015 who underwent cholecystectomy within 15 days of the ultrasonography performed on admittance. Patient data were analyzed retrospectively to compare the ultrasound findings with the pathological findings of the resected gallbladders. Patients were allocated to two groups according to the signs of acute cholecystitis on ultrasonography: group 1 negative and group 2 positive. RESULTS: One hundred and twenty patients were enrolled in the study: 77 in group 1 and 43 in group 2. Similar results were found for the two groups with respect to the pathological diagnosis of acute cholecystitis, i.e., 31.2 % for group 1 and 27.9 % for group 2. Analysis indicated that there was no correlation between the ultrasonography data and pathological findings (p = 0.708). CONCLUSIONS: On the basis of our study, ultrasound findings alone cannot be used to accurately diagnose acute cholecystitis in the setting of acute pancreatitis.

Cdc2-like kinase 2 in the hypothalamus is necessary to maintain energy homeostasis.

OBJECTIVE: To investigate whether the Cdc2-like kinase 2 (CLK2) is expressed in hypothalamic neurons and if it is, whether the hypothalamic CLK2 has a role in the regulation of energy balance. SUBJECTS: Swiss mice on chow or high-fat diet (HFD) and db/db mice on chow diet were used to address the role of CLK2 in the hypothalamus. RESULTS: Hypothalamic CLK2Thr343 phosphorylation, which induces CLK2 activity, is regulated in vivo by refeeding, insulin and leptin, in a PI3K (phosphoinositide 3-kinase)-dependent manner. The reduction of CLK2 expression in the hypothalamus, by chronic pharmacological inhibition with TG003 or by chronic knockdown with small interfering RNA was sufficient to abolish the anorexigenic effect of insulin and leptin, to increase body weight, fat mass, food intake and to decrease energy expenditure in mice on chow. In contrast, CLK2Thr343 phosphorylation in the hypothalamus in response to insulin, leptin or refeeding was impaired in mice on HFD or in db/db mice. Chronic CLK2 inhibition in the hypothalamus was associated with a slight increase in the fasting blood glucose levels, reduction in PEPCK (phosphoenolpyruvate carboxykinase) expression in the liver and enhanced glucose production from pyruvate, suggesting a regulation of hepatic glucose production. Further, overexpressing CLK2 in the mediobasal hypothalami of mice on HFD or in db/db mice by adenovirus partially reversed the obese phenotype. CONCLUSIONS: Thus, our results suggest that protein CLK2 integrates some important hypothalamic pathways, and may be a promising molecule for new therapeutic approaches for obesity and diabetes.

Comparison of simple sucrose and percoll based methodologies for synaptosome enrichment.

Synaptosomes are isolated nerve terminals. They represent an extremely attractive in vitro model system to study synaptic physiology since they preserve morphological and functional characteristics of the synapse. As such they have been used to investigate synaptic dysfunctions associated with neuropathologies like Alzheimer's disease. In the present work two simple methodologies for isolating synaptosomal-enriched fractions were compared for the first time. The starting points of both protocols were rat cortical or hippocampal homogenized tissues that underwent several differential centrifugation steps followed by a final purification of synaptosomal-enriched fractions using either a Percoll gradient or a Sucrose gradient. Comparison of the fractions obtained was carried out, using both biochemical and electron microscopy approaches. In the biochemical analysis the protein levels of pre-synaptic, post-synaptic, nuclear and mitochondrial markers were evaluated. Additional characterization of the synaptosomal-enriched fractions was performed using transmission electron microscopy. In summary, the results indicate that under the conditions tested the Sucrose based protocol is more efficient for the isolation of synaptosomal-enriched fractions from both neuronal tissues, being particularly efficient for hippocampus that is a less abundant brain tissue. Further, the sucrose protocol apparently results in a higher yield of viable synaptosomes suitable for further assays, including structural and functional studies of synapses; making this an attractive procedure to study processes associated with neuropathologies.

Mesenteric lymph node granulomatous lesions in naturally infected wild boar (Sus scrofa) in Portugal--Histological, immunohistochemical and molecular aspects.

Several studies have demonstrated that wildlife reservoirs of mycobacteria are responsible for the maintenance and spreading of the infection to livestock and wildlife counterparts. Recent data report the role of wild boar (Sus scrofa) as a reservoir for Mycobacterium bovis. This study was conducted to evaluate the chronic inflammatory response in the mesenteric lymph nodes (MLN) of wild boar with granulomatous lymphadenitis (n=30). Morphological parameters of the lesions were recorded. The expression of CD3 and CD79α molecules was evaluated by immunohistochemistry. Molecular genotyping and culture to identify mycobacteria were performed. The lesions consisted mainly of stage III and stage IV granulomas. CD3 and CD79α positive cells were observed in 15 (50%) and in 11 (36.6%) MLN, respectively. In these lesions, higher percentages of T lymphocytes were found and a limited number of animals exhibited a tendency for an increased percentage of B lymphocytes. Our results suggest that there are similar percentages and distribution patterns of CD3 and CD79α in the lesions, regardless of the presence of Mycobacterium avium subsp. paratuberculosis (Map), M. bovis or Map-M. bovis co-infection, and confirm that wild boar is both susceptible and could be an important Map and M. bovis wild reservoir in the study area.

RNA sequencing reveals region-specific molecular mechanisms associated with epileptogenesis in a model of classical hippocampal sclerosis.

We report here the first complete transcriptome analysis of the dorsal (dDG) and ventral dentate gyrus (vDG) of a rat epilepsy model presenting a hippocampal lesion with a strict resemblance to classical hippocampal sclerosis (HS). We collected the dDG and vDG by laser microdissection 15 days after electrical stimulation and performed high-throughput RNA-sequencing. There were many differentially regulated genes, some of which were specific to either of the two sub-regions in stimulated animals. Gene ontology analysis indicated an enrichment of inflammation-related processes in both sub-regions and of axonal guidance and calcium signaling processes exclusively in the vDG. There was also a differential regulation of genes encoding molecules involved in synaptic function, neural electrical activity and neuropeptides in stimulated rats. The data presented here suggests, in the time point analyzed, a remarkable interaction among several molecular components which takes place in the damaged hippocampi. Furthermore, even though similar mechanisms may function in different regions of the DG, the molecular components involved seem to be region specific.

VEGFR-2 expression in malignant tumours of the canine mammary gland: a prospective survival study.

Vascular endothelial growth factor receptor-2 (VEGFR-2) is the main receptor activated by vascular endothelial growth factor -A (VEGF-A) to promote tumour angiogenesis. Its clinical prognostic value has not been studied in canine mammary tumours (CMTs). Dogs with mammary cancer were enrolled in a survival study and the immunohistochemical expressions of VEGFR-2 and VEGF-A were analysed and associated with clinicopathological features. VEGFR-2 expression was associated with VEGF immunoreactivity in cancer cells, supporting the presence of an autocrine loop that may be involved in CMTs growth and survival. VEGFR-2 was also expressed by endothelial cells from tumour vasculature and positively associated with stromal matrix metalloproteinase-9 (MMP-9), suggesting the existence of a link between endothelial cells activation and up-regulation of matrix degrading proteins. Carcinosarcomas exhibited high VEGFR-2 expression suggesting that it may be one of the activated molecular pathways in this aggressive histological type and that VEGFR-2 inhibitors may constitute a potential treatment to improve the prognosis of these patients. Both VEGF and VEGFR-2 immunoreactivities were independent of patients' overall survival (OS) and disease-free survival (DFS).