ALS-associated VRK1 R321C mutation causes proteostatic imbalance and mitochondrial defects in iPSC-derived motor neurons.

Vaccinia-related kinase 1 (VRK1) is a gene which has been implicated in the pathological process of a broad range of neurodevelopmental disorders as well as neuropathies, such as Amyotrophic Lateral Sclerosis (ALS). Here we report a family presenting ALS in an autosomal recessive mode of inheritance, segregating with a homozygous missense mutation located in VRK1 gene (p.R321C; Arg321Cys). Proteomic analyses from iPSC-derived motor neurons identified 720 proteins eligible for subsequent investigation, and our exploration of protein profiles revealed significant enrichments in pathways such as mTOR signaling, E2F, MYC targets, DNA repair response, cell proliferation and energetic metabolism. Functional studies further validated such alterations, showing that affected motor neurons presented decreased levels of global protein output, ER stress and downregulation of mTOR signaling. Mitochondrial alterations also pointed to decreased reserve capacity and increased non-mitochondrial oxygen consumption. Taken together, our results present the main pathological alterations associated with VRK1 mutation in ALS.

Exploring Ductility in Dental Ceramics.

Two damage regimes-"brittle" and "ductile"-have been identified in the literature on ceramic grinding, machining, grit blasting, and wear. In the brittle regime, the damage mechanism is essentially crack formation, while in the ductile region, it is quasiplasticity. Onset of the brittle mode poses the greater threat to strength, so it becomes important to understand the mechanics of ductile-brittle thresholds in these materials. Controlled microcontact tests with a sharp indenter are employed to establish such thresholds for a suite of contemporary computer-aided design/computer-aided manufacturing dental ceramics. Plots of flexural strength S versus indentation load P show a steep decline beyond the threshold, consistent with well-established contact mechanics relations. Threshold dimensions occur on a scale of order 1 µm and contact load of order 1 N, values pertinent to practical grit finishing protocols. The ductile side of ceramic shaping is accessed by reducing grit sizes, applied loads, and depths of cut below critical levels. It is advocated that critical conditions for ductile shaping may be most readily quantified on analogous S(P) plots, but with appropriate machining variable (grit size, depths of cut, infeed rate) replacing load P. Working in the ductile region offers the promise of compelling time and cost economies in prosthesis fabrication and preparation.

A sustainable ultra-high strength Fe18Mn3Ti maraging steel through controlled solute segregation and α-Mn nanoprecipitation.

The enormous magnitude of 2 billion tons of alloys produced per year demands a change in design philosophy to make materials environmentally, economically, and socially more sustainable. This disqualifies the use of critical elements that are rare or have questionable origin. Amongst the major alloy strengthening mechanisms, a high-dispersion of second-phase precipitates with sizes in the nanometre range is particularly effective for achieving ultra-high strength. Here, we propose an alternative segregation-based strategy for sustainable steels, free of critical elements, which are rendered ultrastrong by second-phase nano-precipitation. We increase the Mn-content in a supersaturated, metastable Fe-Mn solid solution to trigger compositional fluctuations and nano-segregation in the bulk. These fluctuations act as precursors for the nucleation of an unexpected α-Mn phase, which impedes dislocation motion, thus enabling precipitation strengthening. Our steel outperforms most common commercial alloys, yet it is free of critical elements, making it a new platform for sustainable alloy design.

Boat noise impacts early life stages in the Lusitanian toadfish: A field experiment.

Marine traffic is the most common and chronic source of ocean noise pollution. Despite the evidence of detrimental effects of noise exposure on fish, knowledge about the effects on the critical early life stages - embryos and larvae - is still scarce. Here, we take a natural habitat-based approach to examine potential impacts of boat noise exposure in early life stages in a wild fish population of the Lusitanian toadfish (Halobatrachus didactylus). In-situ experiments were carried out in the Tagus estuary, an estuary with significant commercial and recreational boat traffic. Nests with eggs were exposed to either ambient (control) or boat noise (treatment), for 1 fortnight. Eggs were photographed before being assigned to each treatment, and after exposure, to count number of eggs and/or larvae to assess survival, and sampled to study development and oxidative stress and energy metabolism-related biomarkers. Data concerns 4 sampling periods (fortnights) from 2 years. Results indicate that offspring survival did not differ between treatments, but boat noise induced a detrimental effect on embryos and larvae stress response, and on larvae development. Embryos showed reduced levels of electron transport system (ETS), an energy metabolism-related biomarker, while larvae showed higher overall stress responses, with increased levels of superoxide dismutase (SOD) and DNA damage (oxidative stress related responses), ETS, and reduced growth. With this study, we provided the first evidence of detrimental effects of boat noise exposure on fish development in the field and on stress biomarker responses. If these critical early stages are not able to compensate and/or acclimate to the noise stress later in the ontogeny, then anthropogenic noise has the potential to severely affect this and likely other marine fishes, with further consequences for populations resilience and dynamics.

Failure probability and stress distribution of milled porcelain-zirconia crowns with bioinspired/traditional design and graded interface.

OBJECTIVE: To evaluate the failure probability and stress distribution of traditional and bioinspired porcelain-zirconia milled crowns, with and without silica infiltration (graded zirconia). METHODS: Traditional crown design had a zirconia infrastructure veneered with porcelain; Bioinspired, had a porcelain infrastructure with translucent-zirconia veneer; Graded and Graded Bioinspired crowns had their zirconia layer infiltrated by silica (n = 25). The cameo surface of each crown (porcelain or zirconia) was glazed. The restoration layers were fused by a vitreous connector and the crowns were adhesively cemented to dies. The specimens were then mechanically cycled in a sliding machine using 100 N load at 4 Hz. The specimens were tested until 2 × 106 cycles, and every 0.5 × 106 cycles the crowns were evaluated under stereomicroscopy for the presence of failures. The stress distribution was inspected with Finite Element Analyses. RESULTS: The predominant failure modes for the Traditional and Graded crowns were delamination and cracking, respectively. The Weibull parameters beta and eta were, respectively: Traditional 1.30 and 2.3 × 106 cycles, and Graded 1.95 and 2.3 × 106 cycles. Thus, the Traditional and Graded crowns presented greater susceptibility to failure due to fatigue, while the Bioinspired and Graded Bioinspired crowns showed no fatigue effect using 100N load, showing beta = 1 and eta of approximately 17 × 106 cycles. Also, through finite element analyses, it was verified that the Bioinspired and Graded Bioinspired crowns presented the best stress distribution on both crowns and dental structures. SIGNIFICANCE: Bioinspired and Graded Bioinspired crowns had the lowest failure probability and better stress distribution and may be considered robust long lasting restorations.

Bactericidal Effect of Entomopathogenic Bacterium Pseudomonas entomophila Against Xanthomonas citri Reduces Citrus Canker Disease Severity.

The bacterium Pseudomonas entomophila has been recognized as an exceptional species within the Pseudomonas genus, capable of naturally infecting and killing insects from at least three different orders. P. entomophila ingestion leads to irreversible gut damage resulting from a global blockage of translation, which impairs both immune and tissue repair systems in the insect intestine. In this study we isolated a P. entomophila bacterial strain from soil samples which displayed a strong activity against Xanthomonas citri subsp, citri (Xcc), the etiological agent of citrus canker disease. The antagonism potential of isolated bacteria against Xcc and its ability to reduce citrus canker severity was assessed both ex planta and in planta. Our findings show that pathogenicity assays in Citrus x limonia by pressure infiltration and spray with a mixture of P. entomophila and Xcc leaded to a significant reduction in the number of canker lesions in high susceptible citrus leaves, at 21 days post-infection. To the best of our knowledge this is the first report of antibacterial activity of P. entomophila against a phytopathogenic bacterium. Collective action of P. entomophila factors such as diketopiperazine production and the type 6 secretion system (T6SS) may be involved in this type of biological control of citrus canker. The results suggest that the P. entomophila strain could be a promising biocontrol agent acting directly against Xcc.

Effects of the organophosphorus pesticide Folisuper 600 (methyl parathion) on the heart function of bullfrog tadpoles, Lithobates catesbeianus (Shaw, 1802).

The aim of this work was to evaluate whether the heart function of bullfrog tadpoles (25 Gosner stage) is affected by their acute exposure (48 h) to a sub-lethal concentration (10 µg.L-1) of the active principle of the organophosphorus pesticide Folisuper 600R (methyl parathion - MP). Our results demonstrated that MP causes not only a reduction in tadpoles' cardiac ventricular mass, resulting in a marked reduction in their cardiac twitch force, but also impairs their swimming performance, irrespective of increasing their heart rate. Together, these findings indicate that low and realistic concentration of MP have a negative impact on tadpoles' performance, jeopardizing their survival.

Effects of the organophosphorus pesticide Folisuper 600 (methyl parathion) on the heart function of bullfrog tadpoles, Lithobates catesbeianus (Shaw, 1802)

Abstract The aim of this work was to evaluate whether the heart function of bullfrog tadpoles (25 Gosner stage) is affected by their acute exposure (48 h) to a sub-lethal concentration (10 µg.L1) of the active principle of the organophosphorus pesticide Folisuper 600R (methyl parathion - MP). Our results demonstrated that MP causes not only a reduction in tadpoles cardiac ventricular mass, resulting in a marked reduction in their cardiac twitch force, but also impairs their swimming performance, irrespective of increasing their heart rate. Together, these findings indicate that low and realistic concentration of MP have a negative impact on tadpoles performance, jeopardizing their survival.

Resumo O objetivo do presente estudo foi avaliar se a função cardíaca de girinos de rãs-touro (estágio 25 de Gosner) é afetada pela exposição aguda (48h) a uma concentração sub-letal (10 µg.L1) do princípio ativo do pesticida organofosforado Folisuper 600R (metil paration MP). Nossos resultados demonstraram que o MP ocasionou não apenas uma redução na massa ventricular cardíaca dos girinos, como também provocou uma redução na sua força de contração cardíaca e de seu desempenho natatório, a despeito de ter sido observado um aumento de sua freqüência cardíaca. Conjuntamente, os achados aqui obtidos indicam que uma baixa e realística concentração de MP exerce um impacto negativo sobre o desempenho dos girinos, ameaçando sua sobrevivência.

Effects of the organophosphorus pesticide Folisuper 600 (methyl parathion) on the heart function of bullfrog tadpoles, Lithobates catesbeianus (Shaw, 1802) / Efeitos do pesticida organofosforado Folisuper 600 (metil paration) sobre a função cardíaca de girinos de rã-touro, Lithobates catesbeianus (Shaw, 1802)

Abstract The aim of this work was to evaluate whether the heart function of bullfrog tadpoles (25 Gosner stage) is affected by their acute exposure (48 h) to a sub-lethal concentration (10 µg.L–1) of the active principle of the organophosphorus pesticide Folisuper 600R (methyl parathion - MP). Our results demonstrated that MP causes not only a reduction in tadpoles’ cardiac ventricular mass, resulting in a marked reduction in their cardiac twitch force, but also impairs their swimming performance, irrespective of increasing their heart rate. Together, these findings indicate that low and realistic concentration of MP have a negative impact on tadpoles’ performance, jeopardizing their survival.

Resumo O objetivo do presente estudo foi avaliar se a função cardíaca de girinos de rãs-touro (estágio 25 de Gosner) é afetada pela exposição aguda (48h) a uma concentração sub-letal (10 µg.L–1) do princípio ativo do pesticida organofosforado Folisuper 600R (metil paration – MP). Nossos resultados demonstraram que o MP ocasionou não apenas uma redução na massa ventricular cardíaca dos girinos, como também provocou uma redução na sua força de contração cardíaca e de seu desempenho natatório, a despeito de ter sido observado um aumento de sua freqüência cardíaca. Conjuntamente, os achados aqui obtidos indicam que uma baixa e realística concentração de MP exerce um impacto negativo sobre o desempenho dos girinos, ameaçando sua sobrevivência.

Uso de biomarcador histopatológico em brânquias de Centropomus undecimalis (Bloch, 1972) na avaliação da qualidade da água do Parque Ecológico Laguna da Jansen, São Luís - MA / Using histopathological biomarker in gills of Centropomus undecimalis (Bloch, 1972) in assessing the water quality at Ecological Park Laguna da Jansen, São Luís -MA

Com o objetivo de analisar a intensidade das alterações histológicas em brânquias de Centropomus undecimalis para avaliar a qualidade da água do Parque Ecológico Laguna da Jansen, em São Luís, Estado do Maranhão, coletaram-se exemplares de peixes no período chuvoso de 2012 e amostras de água nos períodos seco de 2011 e chuvoso de 2012 para análises físico-química e bacteriológica. Os resultados mostraram que a maioria dos parâmetros físico-químicos e bacteriológicos estava de acordo com os valores estabelecidos pela Resolução do CONAMA nº 357 de 2005. As alterações histológicas observadas foram: elevação do epitélio lamelar; proliferação excessiva de células dos epitélios filamentar e lamelar causando fusão total ou parcial das lamelas; alteração da estrutura das lamelas; hiperplasia das células mucosas; ruptura do epitélio lamelar; espessamento descontrolado do tecido proliferativo filamentar e lamelar e aneurismas de vários tamanhos. Também foi observada a presença de parasito em alguns exemplares. O Índice de Alteração Histológica (IAH) por indivíduo variou de 2 a 114. O valor médio do IAH obtido foi de 40,3, demonstrando que as brânquias dos indivíduos amostrados da espécie C. undecimalis apresentaram alterações de moderadas para severas no tecido. As alterações histológicas encontradas indicam que esses indivíduos desenvolveram mecanismos de defesa contra a ação de estressores presentes na água da laguna e a um provável desequilíbrio parasito-hospedeiro-ambiente associado à baixa qualidade do ambiente...

Aiming to analyze the intensity of the histological changes in the gills of Centropomus undecimalis to assess the water quality at Ecological Park of the Laguna da Jansen, in São Luís, State of Maranhão, specimens of fish were collected during the rainy season of 2012 and water samples during the dry season in 2011 and rainy season in 2012 for physical-chemical and bacteriological analyses. The results showed that most of the physical-chemical and bacteriological parameters analyzed agreed with the values established by CONAMA Resolution nº 357 of 2005. The histological changes were elevation of the lamellar epithelium, excessive cell proliferation of the filamentum and lamellar epithelium causing total or partial melting of the lamellar; changing the structure of the lamellar, hyperplasia of mucous cells, disruption of the lamellar epithelium, uncontrolled proliferative tissue filamentum thickening and lamellar and aneurysms in various sizes. We noted the presence of parasites in some samples. The Index Histological Amendment (IAH) per individual ranged from 2 to 114. The mean IAH obtained was 40.3, demonstrating that the gills of the individuals of the C. undecimalis species showed moderate to severe changes in the tissue. The histological findings indicate that these individuals have developed defense mechanisms against the action of stressors present in the water of the Laguna da Jansen and a probable parasite-host-environment imbalance associated with the environment's low quality...

Healthcare context and nursing workforce in a main city of Angola.

BACKGROUND: Angola is one of the largest African countries with continuing levels of insecurity, considerable weakness in terms of respect for human rights, destroyed infrastructure and low transparency and social accountability levels. The health system displays gaps and nursing represents the main contingent among human resources in health. AIM: This research aims to understand the healthcare context in Angola from the perspective of Brazilian nurses who were involved in helping their Angolan colleagues. This general view of health services is followed by a description of nursing workforce particularities at a tertiary health service in the province of Luanda. METHODS: Data were extracted from the database of the Global Network of World Health Organization Collaborating Centres for Nursing and Midwifery Development, constructed based on technical visits to Angola in 2009. Information related to health service characteristics was used, focusing on nursing human resource activities at two tertiary, one secondary and one primary health institutions located in the province of Luanda. The study data were analysed through descriptive statistics. FINDINGS: Among the problems the nursing workforce faces, the lack of human, material and financial resources stands out, as well as insufficient professional qualification, excessive work journeys, low remunerations, non-valuation of professionals, leading to unsatisfactory work environments and discouraged human resources. CONCLUSIONS: Nursing in Angola is conquering its professional space. Therefore, regulatory policies are fundamental, defining the rights and obligations of all categories involved, with a view to determining nurses' function in the health team, including respect for and acknowledgement of their role in the community.

Effect of bolus taste on the esophageal transit of patients with stroke.

Stroke is a frequent cause of oropharyngeal dysphagia but may also cause alterations in esophageal motility. The aim of this investigation was to evaluate the effect of bolus taste on the esophageal transit of patients with stroke and controls. Esophageal transit and clearance were evaluated by scintigraphy in 36 patients in the chronic phase of stroke (44-82 years, mean: 63 years) and in 30 controls (33-85 years, mean: 59 years). The patients had a stroke 1-84 months (median: 5.5 months) before the evaluation of esophageal transit. Eight had dysphagia. Each subject swallowed in random order and in the sitting position 5 mL of liquid boluses with bitter (pH=6.0), sour (pH=3.0), sweet (pH=6.9), and neutral (pH=6.8) taste. Transit and clearance duration and the amount of residues were measured in the proximal, middle, and distal esophageal body. There was no difference between patients and controls in esophageal transit or clearance duration. In the distal esophagus, the transit and clearance durations were longer with the sour bolus than with the other boluses in both patients and controls. The amount of residues in the esophageal body was greater in patients than in controls after swallows of the neutral bolus. In control subjects, after swallows of a sour bolus, there was an increase in the amount of residues in the middle and distal esophagus compared with the other boluses. In conclusion, a sour bolus with low pH causes a longer transit and clearance duration in the distal esophageal body. There was no effect of bolus taste or pH on the esophageal transit of patients in the chronic phase of stroke compared with normal volunteers. The longer transit and clearance duration in the distal esophageal body with the sour bolus appears to be a consequence of the low pH of the bolus.

Low somatostatin receptor subtype 2, but not dopamine receptor subtype 2 expression predicts the lack of biochemical response of somatotropinomas to treatment with somatostatin analogs.

OBJECTIVES: To evaluate somatostatin receptor 2A (SSTR2A) and dopamine receptor 2 (DR2) protein expression in somatotropinomas and to relate it to response to somatostatin analogues (SA). DESIGN AND PATIENTS: SSTR2A and DR2 expression was analyzed by immunohistochemistry in 88 somatotropinomas from patients submitted to either pre-surgical or adjuvant SA treatment. Tumors were scored according to percentage of immunostained cells: 0 (< 25%), 1 (25-50%), and 2 (> 50%). Relation between protein expression and response to SA was performed in 66 patients. Response to SA was assessed by percent IGF-I reduction, being considered as an IGF-I per cent reduction higher than 50%. Disease control was also assessed (GH < 1.0 ng/ml and normal IGF-I). RESULTS: SSTR2A and DR2 were expressed in 100% and 98% of tumors, respectively. Biochemical response and disease control rates were 48% and 32%, respectively. Median IGF-I percent reduction after 3 months of SA treatment was lower in the SSTR2A score 0 than in the scores 1 and 2 (p < 0.001, both), and after 6 months in the score 0 than in the score 1 (p = 0.001) and 2 (p < 0.001). Biochemical response and disease control were associated with SSTR2 expression (p < 0.001 and p = 0.004, respectively). A negative predictive value for biochemical response of 100% was found when a SSTR2A expression < 25%of immunostained cells cut-off point was considered. No relation was found between DR2 expression and biochemical response and disease control. CONCLUSION: SSTR2A and DR2 are highly expressed in somatotropinomas. Low SSTR2A, but not DR2, expression is a negative predictive factor to response to SA.

Validation of immunohistochemistry for somatostatin receptor subtype 2A in human somatotropinomas: comparison between quantitative real time RT-PCR and immunohistochemistry.

Somatostatin receptors subtype 2 (SSTR2) expression in somatotropinomas is recognized as a predictor of response to the currently available somatostatin analogs and may be analyzed, mainly, by quantitative RT-PCR or immunohistochemistry (IHC). The former has the advantages of a higher sensitivity and of being quantitative, while the latter, although semi-quantitative, evaluates protein expression and is routinely used in the evaluation of pituitary adenomas. We aimed to evaluate the SSTR2A protein expression in somatotropinomas and to compare it to our previous data regarding mRNA expression, assessed by quantitative real time RTPCR. Thirteen somatotropinomas were analyzed by IHC and the tumors were scored according to percent of immunostained cells: 0 (<25%), 1 (25-50%) and 2 (>50%). SSTR2A immunostaining was present in all but one somatotropinoma, 4 (31%) tumors were classified as score 0, 4 (31%) as score 1, and 5 (38%) as score 2. Median SSTR2 mRNA content was significantly different among the three IHC scores (p=0.036) and was lower in the score 0 than in the score 2 (p=0.016). The finding that there is a positive correlation between RT-PCR and IHC indicates that IHC can be applied in order to assess the SSTR2A content in somatotropinomas.

Estradiol modulates TGF-ß1 expression and its signaling pathway in thyroid stromal cells.

The higher prevalence of thyroid disease in women suggests that estrogen (E2) might be involved in the pathophysiology of thyroid dysfunction. To approach the question of the effect of stromal cells in the modulation of thyroid epithelial cells activity, we established and characterized a homogeneous stromal cell population (TS7 cells) of rat thyroid gland. These fibroblastic cells synthesize the cytoskeleton proteins α-smooth muscle actin and vimentin, produce basement membrane components and express the cytokine transforming growth factor beta 1 (TGF-ß1). Here, we hypothesized that the effects of E2 on follicular thyroid cells are mediated by TGF-ß1 synthesis and secretion by stromal cells (paracrine action). Thus we investigated the effect of E2 on TGF-ß1 synthesis and its signaling pathway in TS7 cells. In addition, we analyzed the role of TGF-ß1 signaling pathway as mediator of TS7-PC CL3 thyroid epithelial cells interactions. We report that TS7 stromal cells expressed α and ß estrogen receptors (ERα and ERß). Further, both isoforms of TGF-ß1 receptors, TGFRI and TGFRII, were also identified in TS7 cells, suggesting that these cells might be a target for this cytokine in vitro. Treatment of TS7 cells with E2 induced both synthesis and secretion of TGF-ß1. This event was followed by phosphorylation of the transcription factor Smad2, a hallmark of TGF-ß1 pathway activation. Co-culture of PC CL3 cells onto TS7 cells monolayers yielded round aggregates of PC CL3 cells surrounded by TS7 cells. TS7 cells induced a decrease in iodide uptake by PC CL3 cells, probably by a mechanism involving TGF-ß1. Moreover, E2 affected synthesis and organization of the extracellular matrix (ECM) components, tenascin C and chondroitin sulfate, in these co-culture cells. Our results point to the TGF-ß1/Smad-2 signaling pathway as a putative target of estrogen actions on thyroid stromal cells and contribute to understanding the interplay between stromal and follicular cells in thyroid physiology.

Effect of bolus volume on proximal esophageal contractions of patients with Chagas' disease and patients with idiopathic achalasia.

Chagas' disease and idiopathic achalasia patients have similar impairment of distal esophageal motility. In Chagas' disease, the contractions occurring in the distal esophageal body are similar after wet or dry swallows. Our aim in this investigation was to evaluate the effect of wet swallows and dry swallows on proximal esophageal contractions of patients with Chagas' disease and with idiopathic achalasia. We studied 49 patients with Chagas' disease, 25 patients with idiopathic achalasia, and 33 normal volunteers. We recorded by the manometric method with continuous water perfusion the pharyngeal contractions 1 cm above the upper esophageal sphincter and the proximal esophageal contractions 5 cm from the pharyngeal recording point. Each subject performed in duplicate swallows of 3-mL and 6-mL boluses of water and dry swallows. We measured the time between the onset of pharyngeal contractions and the onset of proximal esophageal contractions (pharyngeal-esophageal time [PET]), and the amplitude, duration, and area under the curve (AUC) of proximal esophageal contractions. Patients with Chagas' disease and with achalasia had longer PET, lower esophageal proximal contraction amplitude, and lower AUC than controls (P≤ 0.02). In Chagas' disease, wet swallows caused shorter PET, higher amplitude, and higher AUC than dry swallows (P≤ 0.03).There was no difference between swallows of 3- or 6-mL boluses. There was no difference between patients with Chagas' disease and with idiopathic achalasia. We conclude that patients with Chagas' disease and with idiopathic achalasia have a delay in the proximal esophageal response and lower amplitude of the proximal esophageal contractions.

Detecção de Salmonella Albany, Staphylococcus coagulase positivos e micro-organismos mesófilos em carcaças de frango in natura / Decteccion of Salmonella Albany, Coagulase positive Staphylococcus e mesophilic microrganisms in carcasses chicken in natura

Com o objetivo de avaliar as condições higiênico-sanitárias das carcaças de frango in natura comercializadas em mercados públicos do Município de São Luís, MA, foram colhidas quarenta amostras de carcaças de frango e analisadas quanto à pesquisa de Salmonella spp., Staphylococcus coagulase positivos e contagem padrão de micro-organismos mesófilos pelos métodos convencionais. Do total de amostras analisadas, 18 (45%) apresentaram contaminação por Staphylococcus coagulase positivos, 5 (12,5%) estavam contaminadas por Salmonella spp., identificadas como Salmonella Albany. A contagem padrão de micro-organismos mesófilos variou de 103 a 107 UFC/g no alimento analisado. Os resultados obtidos indicam que as condições higiênico-sanitárias das carcaças de frango in natura analisadas são insatisfatórias e que este produto pode ser um importante veículo de toxi-infecção alimentar.

The objective of this study was to evaluate the hygienic-sanitary conditions of in natura broiler carcasses commercialized in the market from São Luiz City, MA. Forty samples were collected and analyzed to research of Salmonella spp., coagulase positive Staphylococcus and of mesophilic microorganisms count. Among the analyzed samples, eighteen (45%) were contaminated with coagulase positive Staphylococcus, five (12,5%) were contaminated with Salmonella sp, being identified Salmonella Albany and counting of 103 to 107 UFC/g of mesophilic microorganisms. The results showed the hygienic-sanitary conditions samples analyzed were unsatisfactory and the consumption of this products can lead to an intoxication.

Carvedilol: just another Beta-blocker or a powerful cardioprotector?

Beta-blockers have been used to treat ischemic heart disease, due to negative chronotropic and inotropic properties, thus inducing a decrease in myocardial consumption of oxygen and nutrients, allowing a better balance between nutritional needs and the supply provided by the coronary blood flow. Recent developments in cell biology allowed us to understand that not all beta-blockers are equal, as their intracellular mechanisms of action can be very different. This paper will focus on carvedilol, a non-selective beta-blocker with alfa-blocker properties, currently used to treat hypertension, heart failure and coronary artery disease. Effects of carvedilol on cardiac mitochondria, their relation to its antioxidant properties, and how these can improve cardiomyocyte resistance to aggression and cardiac function will be discussed. We will begin by depicting the effect of carvedilol on mitochondrial parameters, namely oxidative phosphorylation, calcium homeostasis and energy production. Then we will focus on the mitochondrial permeability transition (MPT) and how the antioxidant properties of carvedilol can be used to minimize oxidative stress, a powerful inducer of MPT. Carvedilol will also be highlighted as an enzyme modulator, focusing on its importance to prevent doxorubicin (DOX) cardiotoxicity. The mitochondrial-related mechanism of cardioprotection involving carvedilol will also be addressed, as we will discuss some clinical pieces of evidence showing the importance of mechanisms previously depicted. In conclusion, based upon its molecular mechanisms of action, carvedilol seems to be a unique beta-blocker. These unique characteristics can help us understand the positive impact of carvedilol on the prognosis of patients with heart disease.

A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers.

AIMS: Detection of Xylella fastidiosa in citrus plants and insect vectors. METHODS AND RESULTS: Chelex 100 resin matrix was successfully standardized allowing a fast DNA extraction of X. fastidiosa. An amplicon of 500 bp was observed in samples of citrus leaf and citrus xylem extract, with and without symptoms of citrus variegated chlorosis, using PCR with a specific primer set indicating the presence of X. fastidiosa. The addition of insoluble acid-washed polyvinylpyrrolidone (PVPP) prior to DNA extraction of insect samples using Chelex 100 resin together with nested-PCR permitted the detection of X. fastidiosa within sharpshooter heads with great sensitivity. It was possible to detect up to two bacteria per reaction. From 250 sharpshooter samples comprising four species (Dilobopterus costalimai, Oncometopia facialis, Bucephalogonia xanthopis and Acrogonia sp.), 87 individuals showed positive results for X. fastidiosa in a nested-PCR assay. CONCLUSIONS: The use of Chelex 100 resin allowed a fast and efficient DNA extraction to be used in the detection of X. fastidiosa in citrus plants and insect vectors by PCR and nested-PCR assays, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The employment of efficient and sensitive methods to detect X. fastidiosa in citrus plants and insect vectors will greatly assist epidemiological studies.

Comparative analyses of the complete genome sequences of Pierce's disease and citrus variegated chlorosis strains of Xylella fastidiosa.

Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X. fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grape-growing region of California. Comparative analyses with a previously sequenced X. fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X. fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X. fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.