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Objetivo. Estimar el efecto de la pandemia de COVID-19 en la provisión, la utilización, la cobertura y la oportunidad de vacunación en niños menores de 5 años. Método. Se realizó un estudio de cohorte retrospectivo con un componente cuantitativo y un componente cualitativo con dos ciclos rápidos de evaluación. En cada ciclo se aplicaron cuestionarios y entrevistas semiestructuradas a miembros del personal de salud y a madres de menores de cinco años. Los instrumentos exploraron la provisión (disponibilidad de insumos y personal) y utilización (demanda y acceso) del programa de inmunizaciones. Para estimar la cobertura de vacunación se realizó un muestreo aleatorio en menores de cinco años en cinco provincias y se analizaron registros administrativos mensuales del programa para el período 2017-2020. Resultados. Aproximadamente 50% de los informantes percibieron una reducción de personal e insumos para el programa. La demanda de atención se redujo en 26% entre el 2020 y el 2021, explicado principalmente por temor al contagio de COVID-19 en unidades de salud. El acceso a la atención del programa se vio afectado por el cambio de horario, espacios ventilados y atención virtual, que no fue bien aceptado por la población. Solo 50% de menores de 1 año tuvieron esquema completo de vacunación y la mayoría de las vacunas no se aplicaron en forma oportuna. Conclusiones. La pandemia de COVID-19 afectó de manera negativa la provisión de vacunas y la utilización del programa de inmunizaciones; en consecuencia, se intensificó la caída de la coberturas y oportunidad de la vacunación, que alcanzaron niveles subóptimos en menores de cinco años en Ecuador. Esta situación aumentó el riesgo de que los niños contraigan enfermedades prevenibles por vacunación.

Context: Ecuador was one of the most affected counties in Latin America during coronavirus disease 2019 (COVID-19). Its health system already faced financing and organizational problems before the pandemic. Aims: To analyze the effects of the pandemic on the provision and utilization of maternal health services in units of the Public Health of Ecuador as perceived by the health personnel. Material and Methods: This is a mixed methods study with two rapid evaluation cycles. The first cycle collected information during January–March and the second one during April–June, 2021. An online questionnaire and semistructured interviews were applied to professionals involved in the management and provision of maternal health programs. Both instruments explored provision and utilization of maternal health services. Questionnaires were processed using SPSS and descriptive statistics. Interviews were exported to Atlas-Ti and were codified using the explored categories. Results: A total of 207 health workers participated in the first cycle, and 200 in the second one. They recognized that during the pandemic, healthcare personnel were reduced by more than 30% due to the presence of risk factors among them as well as the lack of budget for new contracts. The availability of materials and supplies was also reduced by nearly 50%, mainly due to the lack of budget. The demand for maternal healthcare in public health units was reduced because people feared being infected by COVID-19 and nearly 25% goes to the private sector. Conclusions: The pandemic has had a negative effect on the provision as well as utilization of maternal healthcare services. Nevertheless, health professionals implemented diverse strategies to reduce this negative effect on maternal care.

Al hablar o escribir acerca de Daniel Alcides Carrión García surge su sacrificio, pero esta noche del 13 de agosto del 2007 dejando de lado cual fue su contribución a la ciencia, me limitaré a exponerles algunas facetas pocas conocidas y hechos que aún faltan rectificar en relación a su biografía y a su entorno familiar.

La malacoplaquia tiene afinidad especial por el tracto urinario, casi todos los casos en vejiga y aproximadamente el 80-90% tiene cultivos positivos para organismos gramnegativos, la mayoría de los pacientes presentan cierto grado de inmunocompromiso, es importante tenerla en cuenta en pacientes con dichas características aun cuando los estudios de extensión implican otra patología. El porcentaje de falsos positivos en biopsias prostáticas de lesiones categorizadas como PI-RADS (Prostate Imaging-Report and Data System) 5 no es despreciable, sin embargo no es común encontrarlo como diagnóstico diferencial. Paciente de 69 años sin inmunocompromiso, quien presenta inicialmente retención aguda de orina y antígeno prostático específico elevado (8.35 ng/ml), complicado con choque séptico urinario. En la resonancia magnética multiparamétrica se identificó absceso prostático asociado a lesión PI-RADS 5 en zona periférica prostática, biopsiada con diagnóstico histopatológico de malacoplaquia. Es una patología granulomatosa rara, puede ocurrir en cualquier parte del cuerpo, sin embargo la primera vez que se describió la localización prostática fue en 1959, por Carruthers. El conocimiento y sospecha de una entidad inflamatoria, benigna y rara como la malacoplaquia prostática, incluso en el contexto un paciente no inmunocomprometido, con una lesión caracterizada como PI-RADS 5 puede prevenir el mal diagnóstico y sobretratamiento de estos pacientes.

Abstract Inflammatory states and body composition changes are associated with a poor prognosis in many diseases, but their role in coronavirus disease 2019 (COVID-19) is not fully understood. To assess the impact of low skeletal muscle radiodensity (SMD), high neutrophil-to-lymphocyte ratio (NLR) and a composite score based on both variables, on complications, use of ventilatory support, and survival in patients with COVID-19. Medical records of patients hospitalized between May 1, 2020, and July 31, 2020, with a laboratory diagnosis of COVID-19 who underwent computed tomography (CT) were retrospectively reviewed. CT-derived body composition measurements assessed at the first lumbar vertebra level, and laboratory tests performed at diagnosis, were used to calculate SMD and NLR. Prognostic values were estimated via univariate and multivariate logistic regression analyses and the Kaplan–Meier curve. The study was approved by the local Institutional Review Board (CAAE 36276620.2.0000.5404). A total of 200 patients were included. Among the patients assessed, median age was 59 years, 58% were men and 45% required ICU care. A total of 45 (22.5%) patients died. Multivariate logistic analysis demonstrated that a low SMD (OR 2.94; 95% CI 1.13–7.66, P = 0.027), high NLR (OR 3.96; 95% CI 1.24–12.69, P = 0.021) and both low SMD and high NLR (OR 25.58; 95% CI 2.37–276.71, P = 0.008) combined, were associated with an increased risk of death. Patients who had both low SMD and high NLR required more mechanical ventilation (P

Abstract Hepatocellular carcinoma (HCC) is amongst the cancers with highest mortality rates and is the most common malignancy of the liver. Early detection is vital to provide the best treatment possible and liquid biopsies combined with analysis of circulating tumour DNA methylation show great promise as a non-invasive approach for early cancer diagnosis and monitoring with low false negative rates. To identify reliable diagnostic biomarkers of early HCC, we performed a systematic analysis of multiple hepatocellular studies and datasets comprising > 1500 genome-wide DNA methylation arrays, to define a methylation signature predictive of HCC in both tissue and cell-free DNA liquid biopsy samples. Our machine learning pipeline identified differentially methylated regions in HCC, some associated with transcriptional repression of genes related with cancer progression, that benchmarked positively against independent methylation signatures. Combining our signature of 38 DNA methylation regions, we derived a HCC detection score which confirmed the utility of our approach by identifying in an independent dataset 96% of HCC tissue samples with a precision of 98%, and most importantly successfully separated cfDNA of tumour samples from healthy controls. Notably, our risk score could identify cell-free DNA samples from patients with other tumours, including colorectal cancer. Taken together, we propose a comprehensive HCC DNA methylation fingerprint and an associated risk score for detection of HCC from tissue and liquid biopsies.

Abstract In patients with diabetes, it has been suggested that physical exercise may reduce albuminuria and the progression of renal disease. However, the molecular mechanism by which physical exercise protects the kidney in diabetes remains poorly understood. The aim of the present study was to determine the contribution of muscle irisin secretion induced by aerobic physical exercise with the subsequent activation of AMPK for kidney protection under diabetic conditions. Aerobic physical exercise in rats protected the kidney in streptozotocin-induced diabetes. It reduced albuminuria, glomerular hypertrophy, and glomerular expression of collagen IV and fibronectin, as well as markers of kidney inflammation, when compared to sedentary diabetic rats. These effects were associated with elevation in muscle FNDC5/irisin and activity of AMPK in the diabetic kidney. However, the beneficial effects of exercise were lost when the diabetic rats were treated with CycloRGDyK, that in the bone it has been described as an irisin receptor blocker. In cultured human tubular (HK-2) cells, treatment with recombinant irisin counteracted the effect of high glucose in a dose-dependent manner. Irisin, per se, also activated AMPK in HK-2 cells. It is concluded that in diabetes, the renal protective effect of exercise may be mediated by the irisin/AMPK pathway.

The quality of drug products may be affected from manufacture to dispensing, particularly at high temperature and humidity as in Mauritania. This country is not included in the World Health Organization reports on poor quality products due to the lack of a qualified laboratory and monitoring system. Ensuring the quality of medicine is even more relevant in the case of diseases such as Tuberculosis, due to its high prevalence, complex treatment and continuous bacterial resistance. The aim was to develop a monitoring system to assess the quality of antituberculosis drugs products, by the substandard detection based on European and United States Pharmacopeial recommendations regarding quality control. In addition to studying the influence of accelerated storage conditions (40 ± 2°C/75 ± 5% relative humidity) on their qualities and comparing the dissolution profiles to contrast the quality. 18 antituberculosis drug products were taken from Europe and Mauritania, and quality was studied through visual inspection and according to the compliance of the mass uniformity, uniformity of dosage units, dissolution, disintegration and friability pharmacopeial tests. Furthermore, a dissolution profile comparison was carried out to examine quality. A stability study was conducted to assess the influence of climatic conditions on the content and the dissolved amount of the active pharmaceutical ingredients, which were determined by an ultra-performance liquid chromatography system. As result, 69.3% of 13 Mauritanian formulations had a substandard quality mainly due to non-compliance with the test for friability or content uniformity of these medicines. All European drug products complied with pharmacopeia specifications. In addition, storage conditions affected the dissolution rate of ethambutol and the uniformity of the 4 antituberculosis combination drug products.

Cells recognize both foreign and host-derived double-stranded RNA (dsRNA) via a signaling pathway that is usually studied in the context of viral infection. It has become increasingly clear that the sensing and handling of endogenous dsRNA is also critical for cellular differentiation and development. The adenosine RNA deaminase, ADAR1, has been implicated as a central regulator of the dsRNA response, but how regulation of the dsRNA response might mediate cell fate during injury and whether such signaling is cell intrinsic remain unclear. Here, we show that the ADAR1-mediated response to dsRNA was dramatically induced in 2 distinct injury models of gastric metaplasia. Mouse organoid and in vivo genetic models showed that ADAR1 coordinated a cell-intrinsic, epithelium-autonomous, and interferon signaling–independent dsRNA response. In addition, dsRNA accumulated within a differentiated epithelial population (chief cells) in mouse and human stomachs as these cells reprogrammed to a proliferative, reparative (metaplastic) state. Finally, chief cells required ADAR1 to reenter the cell cycle during metaplasia. Thus, cell-intrinsic ADAR1 signaling is critical for the induction of metaplasia. Because metaplasia increases cancer risk, these findings support roles for ADAR1 and the response to dsRNA in oncogenesis.

Plants have developed various mechanisms to respond specifically to each biotrophic attack. It has been shown that the electrical signals emitted by plants are associated with herbivory stress responses and can lead to the activation of multiple defences. Bt cotton is a genetically modified pest-resistant plant that produces an insecticide from Bacillus thuringiensis (Bt) to control Lepidopteran species. Surprisingly, there is no study-yet, that characterizes the signalling mechanisms in transgenic cotton plants attacked by non-target insects, such as aphids. In this study, we characterized the production of electrical signals on Bt and non-Bt cotton plants infested with Aphis gossypii and, in addition, we characterized the dispersal behaviour of aphids to correlate this behaviour to plant signalling responses. Electrical signalling of the plants was recorded with an extracellular measurement technique. Impressively, our results showed that both Bt and non-Bt cotton varieties, when attacked by A. gossypii, emitted potential variation-type electrical signals and clearly showed the presence of distinct responses regarding their perception and the behaviour of aphids, with evidence of delay, in terms of signal amount, and almost twice the amount of Cry1F protein was observed on Bt cotton plants at the highest density of insects/plant. We present in our article some hypotheses that are based on plant physiology and insect behaviour to explain the responses found on Bt cotton plants under aphid stress.

Objective To analyse how non-adherence to prescribed treatments might be prevented, screened, assessed and managed in people with rheumatic and musculoskeletal diseases (RMDs).Methods An overview of systematic reviews (SR) was performed in four bibliographic databases. Research questions focused on: (1) effective interventions or strategies, (2) associated factors, (3) impact of shared decision making and effective communication, (4) practical things to prevent non-adherence, (5) effect of non-adherence on outcome, (6) screening and assessment tools and (7) responsible healthcare providers. The methodological quality of the reviews was assessed using AMSTAR-2. The qualitative synthesis focused on results and on the level of evidence attained from the studies included in the reviews.Results After reviewing 9908 titles, the overview included 38 SR on medication, 29 on non-pharmacological interventions and 28 on assessment. Content and quality of the included SR was very heterogeneous. The number of factors that may influence adherence exceed 700. Among 53 intervention studies, 54.7% showed a small statistically significant effect on adherence, and all three multicomponent interventions, including different modes of patient education and delivered by a variety of healthcare providers, showed a positive result in adherence to medication. No single assessment provided a comprehensive measure of adherence to either medication or exercise.Conclusions The results underscore the complexity of non-adherence, its changing pattern and dependence on multi-level factors, the need to involve all stakeholders in all steps, the absence of a gold standard for screening and the requirement of multi-component interventions to manage it.

The centrosome is composed of two centrioles surrounded by a microtubule-nucleating pericentriolar material (PCM). Although centrioles are known to regulate PCM assembly, it is less known whether and how the PCM contributes to centriole assembly. Here we investigate the interaction between centriole components and the PCM by taking advantage of fission yeast, which has a centriole-free, PCM-containing centrosome, the SPB. Surprisingly, we observed that several ectopically-expressed animal centriole components such as SAS-6 are recruited to the SPB. We revealed that a conserved PCM component, Pcp1/pericentrin, interacts with and recruits SAS-6. This interaction is conserved and important for centriole assembly, particularly its elongation. We further explored how yeasts kept this interaction even after centriole loss and showed that the conserved calmodulin-binding region of Pcp1/pericentrin is critical for SAS-6 interaction. Our work suggests that the PCM not only recruits and concentrates microtubule-nucleators, but also the centriole assembly machinery, promoting biogenesis close by.

Fusarium kuroshium is the fungal symbiont associated with the ambrosia beetle Euwallacea kuroshio, a plague complex that attacks avocado, among other hosts, causing a disease named Fusarium dieback (FD). However, the contribution of F. kuroshium to the establishment of this disease remains unknown. To advance the understanding of F. kuroshium pathogenicity, we profiled its exo-metabolome through metabolomics tools based on accurate mass spectrometry. We found that F. kuroshium can produce several key metabolites with phytotoxicity properties and other compounds with unknown functions. Among the metabolites identified in the fungal exo-metabolome, fusaric acid (FA) was further studied due to its phytotoxicity and relevance as a virulence factor. We tested both FA and organic extracts from F. kuroshium at various dilutions in avocado foliar tissue and found that they caused necrosis and chlorosis, resembling symptoms similar to those observed in FD. This study reports for first-time insights regarding F. kuroshium associated with its virulence, which could lead to the potential development of diagnostic and management tools of FD disease and provides a basis for understanding the interaction of F. kuroshium with its host plants.

La eficacia de la terapia de resincronización (RSC) en pacientes con insuficiencia cardíaca avanzada y asincronismo sistólico está ampliamente demostrada, aun cuando sus resultados se ven limitados por la variabilidad anatómica del sistema venoso coronario, el cual no siempre permite una adecuada colocación del electrodo ventricular izquierdo (VI). El asincronismo ventricular agudo es un factor de mal pronóstico que está presente en un considerable número de pacientes con indicación de cirugía cardíaca y disfunción sistólica. La cirugía con circulación extracorpórea (CEC) proporciona el escenario adecuado para implantar el electrodo epicárdico en la base de la pared libre del VI, permitiendo realizar una RSC epicárdica completa concomitante, diferida o profiláctica. En la mayoría de los casos de cardiopatía isquémica con asincronismo del VI, la revascularización (RVC) miocárdica ni revierte el asincronismo ni mejora la función sistólica. Tras la cirugía, la indicación de RSC se mantiene en la mayoría de los casos. La implantación epicárdica del sistema de RSC asociada a la RVC miocárdica facilita el manejo del paciente en el postoperatorio inmediato, mejora de forma significativa la función sistólica del VI y aumenta la supervivencia. La insuficiencia mitral funcional (IMF) del ventrículo dilatado puede precisar de un enfoque terapéutico múltiple por su frecuente asociación con el asincronismo papilar y que incluye RVC miocárdica, reparación mitral, RSC e incluso remodelado ventricular. La insuficiencia mitral funcional isquémica moderada sometida a RVC miocárdica solo mejora en los casos con presencia de miocardio viable y ausencia de asincronismo papilar. La determinación preoperatoria de estos hallazgos indica que la RVC puede tener que acompañarse de RSC para restablecer el sincronismo interpapilar. La indicación de RSC asociada a la cirugía de la cardiopatía isquémica ha demostrado ser una estrategia quirúrgica de probada eficacia y durabilidad. El implante de electrodos epicárdicos de forma concomitante debe realizarse cuando existe indicación de RSC o de un upgrading.

El manejo perioperatorio, tanto tras la circulación extracorpórea (CEC) como en el postoperatorio inmediato, es complejo y cambiante, pudiendo requerir de diferentes niveles de soporte hemodinámico, administración de volumen, fármacos vasoactivos e inotrópicos, estimulación eléctrica optimizada e incluso asistencia cardiocirculatoria mecánica. Estudios recientes han mostrado los efectos deletéreos de la administración de fármacos inotrópicos que afectan al equilibrio consumo de oxígeno hístico/aporte. La optimización de la estimulación eléctrica temporal para mejorar la función ventricular y minimizar las arritmias puede proporcionar estabilidad hemodinámica, y ha demostrado ser beneficiosa selectivamente, aun cuando en general sigue siendo una herramienta infrautilizada por su especificidad y desconocimiento de las distintas estrategias y técnicas de estimulación a emplear. En esta revisión se proporciona una información actualizada de la estimulación eléctrica perioperatoria optimizada, que sirva de guía para su utilización rutinaria como parte del arsenal terapéutico perioperatorio.

OBJETIVO: Avaliar os resultados da retificação do eixo mecânico dos membros inferiores por meio da correção aguda de deformidade distal do fêmur usando um tipo de fixação externa híbrida dos fragmentos. MÉTODOS: Quinze joelhos de 11 pacientes com deformidade na extremidade distal do fêmur foram submetidos à correção do ângulo frontal do joelho por apresentarem deformidades em varo ou valgo, sem desvios no plano sagital. A técnica incluiu planejamento pré-operatório, osteotomia percutânea na metáfise distal do fêmur, correção aguda da deformidade angular associada à translação, e fixação da osteotomia por fixador externo híbrido com montagem modificada. A avaliação radiográfica foi realizada em períodos intermediários ao longo dos seis meses do período pós-operatório, com acompanhamento clínico médio de 17 meses. RESULTADOS: Todas as osteotomias consolidaram-se, em tempo médio de três meses. Em apenas um caso houve complicação clínica significativa representada por fratura após a retirada do fixador, o que levou a nova colocação. O ângulo mecânico femoral distal lateral (mLDFA) retornou aos valores normais em nove de 15 joelhos, com média de 71,5%. Em quatro extremidades, desvios residuais leves do eixo mecânico foram causados por deformidade tibial e em uma por deformidade em varo da extremidade proximal do fêmur. CONCLUSÃO: A técnica de correção do desvio do segmento distal do fêmur, por osteotomia distal compensada pela translação do fragmento, foi adequada para restaurar o eixo mecânico do membro inferior e o fixador externo híbrido com a montagem modificada foi eficiente para manter as correções agudas obtidas no momento da cirurgia e propiciar a consolidação óssea.OBJECTIVE: To evaluate the results achieved with the correction of the mechanical axis of lower limbs through acute correction of a distal femur deformity using a hybrid type of external fixation to fixate the fragments. METHODS: 15 knees of 11 patients with deformity in the distal femur extremity were submitted to correction of the frontal angle of the knee because they had either varus or valgus deformity, without displacement in the sagittal plan. The technique involved pre-operative planning, percutaneous osteotomy in the distal femoral metaphysis, acute correction of the angular deformity associated to translation, fixation of the osteotomy using a hybrid external fixator with a modified assembly. Radiographic evaluation was performed in intermediary periods along the six-month period after surgery, with mean clinical follow-up of 17 months. RESULTS: Union was achieved in all of the osteotomies in a mean time of three months. Only one case presented a significant clinical complication represented by a fracture after the fixator was removed, and a new fixator had to be placed. The lateral distal femoral mechanical angle (mLDFA) returned to normal values in 9 of the 15 knees, with a mean of 71.5%. In four extremities, slight residual displacement of the mechanical axis was caused by tibial deformity and one by a varus deformity in the proximal femur extremity. CONCLUSION: The technique of correcting the displacement in the distal segment of the femur using distal osteotomy offset by the fragment translation was adequate to recover the mechanical axis of the lower limb, and the hybrid external fixator with the modified assembly was efficient to maintain the acute corrections achieved by the surgery and to provide for bone union.

OBJETIVO: Analisar, por meio da histomorfometria, o efeito do alendronato de sódio sobre o trabeculado ósseo de ratos, quando administrado simultaneamente com imobilização gessada. MÉTODOS: Foram utilizados quatro grupos com cinco fêmeas de ratos Wistar: 1) imobilizado; 2) não imobilizado + alendronato; 3) imobilizado + alendronato; e 4) controle. A imobilização foi feita com gesso pelvipodálico aplicado até o membro posterior direito e o alendronato foi administrado em doses semanais. O período de observação foi de 28 dias e realizada histomorfometria da metáfise proximal da tíbia, com análise do número de trabéculas, volume ósseo, espessura trabecular e separação trabecular. RESULTADOS: O grupo imobilizado apresentou volume ósseo menor que os demais grupos. Os animais que receberam alendronato semanal, tanto imobilizados, quanto não imobilizados, apresentaram volume ósseo maior que o controle. A espessura trabecular no grupo imobilizado foi menor do que nos grupos controle e não imobilizado que recebeu alendronato, mas não apresentou diferença significativa em relação ao imobilizado com alendronato. O grupo imobilizado apresentou separação trabecular maior que os demais grupos. Os grupos não imobilizado, sem imobilização que recebeu alendronato e imobilizado que recebeu alendronato apresentaram aumento no número de trabéculas em relação ao grupo imobilizado. CONCLUSÃO: A imobilização empregada efetivamente levou à osteopenia, verificada pela diminuição de todos os principais parâmetros histomorfométricos estudados. Estas alterações foram prevenidas pela administração concomitante de alendronato sódico, exceto com relação à espessura trabecular. O alendronato de sódio foi capaz de aumentar os parâmetros morfométricos, mesmo em animais não imobilizados.OBJECTIVE: Using histomorphometric means to analyze the effect of alendronate sodium on the bone trabeculate of rats administered concomitantly with cast immobilization. METHODS: Female Wistar rats were distributed in four groups with five animals each: 1) cast-immobilized; 2) no immobilization + alendronate; 3) cast-immobilized + alendronate; and 4) control. Immobilization was done with pelvipodalic cast applied till the right hind limb and alendronate was administered in weekly doses. The observation period was 28 days and histomorphometric evaluations were performed in the proximal tibial metaphysis, analyzing the number of trabeculae, bone volume, trabecular thickness, and trabecular separation. RESULTS: The immobilized group presented less bone volume than the other groups. The animals receiving alendronate every week, whether or not immobilized, presented a greater bone volume than the control group. Trabecular thickness in the immobilized group was less than in the control and in the non-immobilized groups that received alendronate, but had no significant difference when compared to the immobilized with alendronate group. The immobilized group presented greater trabecular separation than the other groups. In the non-immobilized groups, the non-immobilized group that received alendronate and the immobilized group that received alendronate presented an increased number of trabeculae when compared to the immobilized group. CONCLUSION: The immobilization used led to osteopenia, as confirmed by the decrease in all of the main histomorphometric parameters studied. Such changes were prevented with the concomitant administration of alendronate sodium, exception being made to the trabecular thickness. Alendronate sodium was able to increase morphometric parameters, even in non-immobilized animals.

Acrostichum is considered today an opportunistic fern in disturbed areas, which indicates the first stages of colonisation of such zones. However, in the fossil record, Acrostichum appears related to fluvio-lacustrine environments, freshwater marshes and mangrove deposits. We report here for first time fossil evidence of Acrostichum that reveals a pioneering behaviour of this fern in the colonisation of perturbed communities in Europe, which corroborates previous assumptions about the paleobiology of Acrostichum. Plant remains were collected from the Chattian (late Oligocene) La Val fossil site (Estadilla, Huesca, northeastern Spain) belonging to the Sariñena Formation, which mainly embraces crevasse splays, levees and floodplain deposits. Evidence shows that Acrostichum grew within the levee's vegetal community or close to/on the river banks as well as on floodplain areas and closer to/on the shores of ephemeral ponds. But most importantly, the observed co-existence of Equisetum and Acrostichum remains in the same beds indicates that such strata represent short-lived inundated terrains, e.g., floodplains where the water table was temporarily stagnant. Evidence shows wetland environments dominated by pioneering taxa, implying a pioneering role for Acrostichum during the late Oligocene in the Iberian Peninsula.

Bone marrow-derived cells were demonstrated to improve organ function, but the lack of cell retention within injured organs suggests that the protective effects are due to factors released by the cells. Herein, we tested cell therapy using early outgrowth cells (EOCs) or their conditioned media (CM) to protect the retina of diabetic animal models (type 1 and type 2) and assessed the mechanisms by in vitro study. Control and diabetic (db/db) mice (8 weeks of age) were randomized to receive a unique intravenous injection of 5×105EOCs or 0.25 ml thrice weekly tail-vein injections of 10x concentrated CM and Wystar Kyoto rats rendered diabetic were randomized to receive 0.50 ml thrice weekly tail-vein injections of 10x concentrated CM. Four weeks later, the animals were euthanized and the eyes were enucleated. Rat retinal Müller cells (rMCs) were exposed for 24 h to high glucose (HG), combined or not with EOC-conditioned medium (EOC-CM) from db/m EOC cultures. Diabetic animals showed increase in diabetic retinopathy (DR) and oxidative damage markers; the treatment with EOCs or CM infusions significantly reduced this damage and re-established the retinal function. In rMCs exposed to diabetic milieu conditions (HG), the presence of EOC-CM reduced reactive oxygen species production by modulating the NADPH-oxidase 4 system, thus upregulating SIRT1 activity and deacetylating Lys-310-p65-NFκB, decreasing GFAP and VEGF expressions. The antioxidant capacity of EOC-CM led to the prevention of carbonylation and nitrosylation posttranslational modifications on the SIRT1 molecule, preserving its activity. The pivotal role of SIRT1 on the mode of action of EOCs or their CM was also demonstrated on diabetic retina. These findings suggest that EOCs are effective as a form of systemic delivery for preventing the early molecular markers of DR and its conditioned medium is equally protective revealing a novel possibility for cell-free therapy for the treatment of DR.

Barrett's esophagus is the major risk factor for esophageal adenocarcinoma. It has a low but non-neglectable risk, high surveillance costs and no reliable risk stratification markers. We sought to identify early biomarkers, predictive of Barrett's malignant progression, using a meta-analysis approach on gene expression data. This in silico strategy was followed by experimental validation in a cohort of patients with extended follow up from the Instituto Português de Oncologia de Lisboa de Francisco Gentil EPE (Portugal). Bioinformatics and systems biology approaches singled out two candidate predictive markers for Barrett's progression, CYR61 and TAZ. Although previously implicated in other malignancies and in epithelial-to-mesenchymal transition phenotypes, our experimental validation shows for the first time that CYR61 and TAZ have the potential to be predictive biomarkers for cancer progression. Experimental validation by reverse transcriptase quantitative PCR and immunohistochemistry confirmed the up-regulation of both genes in Barrett's samples associated with high-grade dysplasia/adenocarcinoma. In our cohort CYR61 and TAZ up-regulation ranged from one to ten years prior to progression to adenocarcinoma in Barrett's esophagus index samples. Finally, we found that CYR61 and TAZ over-expression is correlated with early focal signs of epithelial to mesenchymal transition. Our results highlight both CYR61 and TAZ genes as potential predictive biomarkers for stratification of the risk for development of adenocarcinoma and suggest a potential mechanistic route for Barrett's esophagus neoplastic progression.

La infección de los electrodos de marcapasos endocavitarios es una complicación grave que puede dar lugar a endocarditis, embolia pulmonar y septicemia. El tratamiento de elección consiste en el explante completo del dispositivo ya sea quirúrgico o percutáneo. Presentamos a continuación nuestra experiencia en la extracción de electrodos de marcapasos infectados mediante cirugía cardíaca abierta con circulación extracorpórea. Pacientes y métodos: Se realizó un estudio retrospectivo sobre un total de 27 pacientes consecutivos con infección de electrodos de marcapasos extraídos bajo circulación extracorpórea entre 2000–2006. Se analizaron las variables clínicas, microbiológicas y quirúrgicas. Resultados: La edad media de los pacientes fue de 66,7 ± 11 años, con un 74% de varones. Todos los pacientes habían sido tratados antes de la intervención con antibióticos y en el 55,5% se había extraído parcialmente el sistema de marcapasos. El germen más frecuente fue el S. epidermidis, seguido por el S. aureus. La ecocardiografía transtorácica y/o transesofágica preoperatoria identificó vegetaciones en el 93,6% de los casos. La mortalidad hospitalaria fue del 3,7%. Presentaron insuficiencia renal postoperatoria que precisó hemodiafiltración el 25,9% e insuficiencia respiratoria con ventilación mecánica prolongada el 22%. En el seguimiento postoperatorio todos los pacientes se encuentran libres de recurrencia de endocarditis. Conclusiones: El explante bajo circulación extracorpórea del sistema de marcapasos en pacientes con endocarditis sobre electrodos es una técnica efectiva. Aunque presenta una morbimortalidad significativa, en ocasiones es el único método capaz de extraer por completo el material infectado, por lo que constituye una herramienta terapéutica de primera línea junto con los procedimientos percutáneos.

Parafusos confeccionados de osso são pouco estudados na literatura. Aspectos estruturais, mecânicos e relativos a osteointegração e osteoindução são importantes tópicos de pesquisa a serem abrangidos previamente ao uso in vivo de implantes de osso. Entretanto, o fator inicial a ser avaliado é a usinabilidade do tecido ósseo. Outro ponto de relevância é a verificação da praticabilidade de confeccionar parafusos ósseos de dimensões pré-estabelecidas. Nestas pesquisas, os parafusos foram confeccionados a partir de amostras de osso cortical retiradas de forma padronizada da porção médio-diafisária da tíbia bovina. A usinagem do osso foi realizada em torno horizontal com uso de duas ferramentas: bits de aço e rebolo de ponta montada de óxido de alumínio, produzindo-se 22 parafusos. A avaliação das dimensões dos parafusos ósseos foi realizada em projetor de perfil, tendo-se como referência 10 parafusos metálicos. De forma geral a análise metrológica não mostrou variações significativas dentro do grupo dos parafusos ósseos e metálicos. Os principais problemas dimensionais encontrados nos dois grupos de parafusos foram: ângulos alfa muito elevados nos implantes ósseos e os similares metálicos apresentaram um diâmetro de corpo com valor muito abaixo do esperado. Concluímos que o tecido ósseo foi usinável e a confecção de parafusos de dimensões pré-estabelecidas, mostrou-se possível no osso.Screws made of bone are little studied in literature. Structural, mechanical and osseointegration/ osteoinduction-related aspects are important research topics to be addressed prior to the use of in vivo bone grafts. However, the first issue that should be addressed is the machineability of bone tissues. Another relevant issue is regarding the feasibility of building bone screws in pre-determined dimensions. In those researches, screws were made of cortical bone samples removed from the medial-diaphyseal portion of bovine tibiae in a standardized fashion. Bone machining was performed in a horizontal lathe by using two pieces of tool: steel bits and aluminum-oxide tip-mounted grindstone, producing 22 screws. The evaluation of bone screws' dimensions was performed in a profile projector, taking 10 metal screws as reference. In general, the metrological analysis showed no significant changes within bone and metal screws groups. Major dimensional problems found in both screw groups were: very high alpha angles in bone grafts and similar metal grafts presenting a body diameter in a much lower value than expected. We concluded that bone tissue is machineable and the production of screws with pre-defined dimensions was shown to be possible in the bone.

Aphids cause significant damage to crop plants. Studies regarding predator-prey relationships in fennel (Foeniculum vulgare Mill.) and cotton (Gossypium hirsutum L.) crops are important for understanding essential ecological interactions in the context of intercropping and for establishing pest management programs for aphids. This study evaluated the association among Hyadaphis foeniculi (Passerini) (Hemiptera: Aphididae), Aphis gossypii Glover (Hemiptera: Aphididae) and Cycloneda sanguinea (L.) (Coleoptera: Coccinellidae) in cotton with coloured fibres, fennel and cotton intercropped with fennel. Association analysis was used to investigate whether the presence or absence of prey and predator species can indicate possible interactions between aphids and ladybugs. Significant associations among both apterous and alate H. foeniculi and C. sanguinea were observed in both the fennel and fennel-cotton intercropping systems. The similarity analysis showed that the presence of aphids and ladybugs in the same system is significantly dependent on the type of crop. A substantial amount of evidence indicates that the presence of the ladybug C. sanguinea, is associated with apterous or alate A. gossypii and H. foeniculi in fennel-cotton intercropping system. We recommend that future research vising integrated aphid management taking into account these associations for take decisions.

Fingerprinting information can be used to elucidate in a robust manner the genetic structure of germplasm collections, allowing a more rational and fine assessment of genetic resources. Bayesian model-based approaches are nowadays majorly preferred to infer genetic structure, but it is still largely unresolved how marker sets should be built in order to obtain a robust inference. The objective was to evaluate, in Pyrus germplasm collections, the influence of the SSR marker set size on the genetic structure inferred, also evaluating the influence of the criterion used to select those markers. Inferences were performed considering an increasing number of SSR markers that ranged from just two up to 25, incorporated one at a time into the analysis. The influence of the number of SSR markers used was evaluated comparing the number of populations and the strength of the signal detected, and also the similarity of the genotype assignments to populations between analyses. In order to test if those results were influenced by the criterion used to select the SSRs, several choosing scenarios based on the discrimination power or the fixation index values of the SSRs were tested. Our results indicate that population structure could be inferred accurately once a certain SSR number threshold was reached, which depended on the underlying structure within the genotypes, but the method used to select the markers included on each set appeared not to be very relevant. The minimum number of SSRs required to provide robust structure inferences and adequate measurements of the differentiation, even when low differentiation levels exist within populations, was proved similar to that of the complete list of recommended markers for fingerprinting. When a SSR set size similar to the minimum marker sets recommended for fingerprinting it is used, only major divisions or moderate (FST>0.05) differentiation of the germplasm are detected.

The FOXP3 gene encodes for a protein (Foxp3) involved in the development and functional activity of regulatory T cells (CD4+/CD25+/Foxp3+), which exert regulatory and suppressive roles over the immune system. After allogeneic stem cell transplantation, regulatory T cells are known to mitigate graft versus host disease while probably maintaining a graft versus leukemia effect. Short alleles (≤(GT)15) for the (GT)n polymorphism in the promoter/enhancer of FOXP3 are associated with a higher expression of FOXP3, and hypothetically with an increase of regulatory T cell activity. This polymorphism has been related to the development of auto- or alloimmune conditions including type 1 diabetes or graft rejection in renal transplant recipients. However, its impact in the allo-transplant setting has not been analyzed. In the present study, which includes 252 myeloablative HLA-identical allo-transplants, multivariate analysis revealed a lower incidence of grade III-IV acute graft versus host disease (GVHD) in patients transplanted from donors harboring short alleles (OR = 0.26, CI 0.08-0.82, p = 0.021); without affecting chronic GVHD or graft versus leukemia effect, since cumulative incidence of relapse, event free survival and overall survival rates are similar in both groups of patients.

Hyadaphis foeniculi (Passerini) (Hemiptera: Aphididae) is a cosmopolitan species and the main pest of fennel in northeastern Brazil. Understanding the relationship between temperature variations and the population growth rates of H. foeniculi is essential to predict the population dynamics of this aphid in the fennel crop. The aim of this study was to measure the effect of constant temperature on the adult prereproductive period and the life table fertility parameters (infinitesimal increase ratio (rm), gross reproduction rate (GRR), net reproduction rate (R0), finite increase ratio (λ), generation time (GT), the time required for the population to double in the number of individuals (DT), and the reproduction value (RVx)) of the fennel pest H. foeniculi. The values of lx (survival of nymphs at age x) increased as the temperature rose from 15 to 28°C and fell at 30°C, whereas mx (number of nymphs produced by each nymph of age x) increased from 15 to 25°C and fell at 28 and 30°C. The net reproduction rates (R0) of populations of H. foeniculi increased with temperature and ranged from 1.9 at 15°C to 12.23 at 28°C for each generation. The highest population increase occurred with the apterous aphids at 28°C. The rate of population increase per unit time (rm) (day) ranged from 0.0033 (15°C) to 0.1995 (28°C). The highest values of rm were recorded at temperatures of 28°C and 30°C. The rm values were a good fit to the models tested, with R2 > 0.91 and R2adj > 0.88. The models tested (Davidson, Sharpe and DeMichele modified by Schoolfield et al., Logan et al., Lamb, and Briere et al.) were very good fits for the rm values observed, with R2 > 0.91 and R2adj > 0.88. The only exception was the Davidson model. Of the parameters studied, the reproductive capacity was higher in the apterous aphids, with the unique exception of daily fecundity at 28°C, which was higher in the alate aphids of H. foeniculi. Parameters relating to the age-specific fertility table for H. foeniculi were heavily influenced by temperature, with the highest biotic potential and population growth capacity found at 34°C. Therefore, the results obtained in this study could be of practical significance for predicting outbreaks of fennel aphids and improving the management of this aphid in fennel crops.

Metabolic syndrome is a cluster of metabolic risk factors such as obesity, diabetes and cardiovascular diseases. Mitochondria is the main site of ATP production and its dysfunction leads to decreased oxidative phosphorylation, resulting in lipid accumulation and insulin resistance. Our group has demonstrated that kinins can modulate glucose and lipid metabolism as well as skeletal muscle mass. By using B2 receptor knockout mice (B2R-/-) we investigated whether kinin action affects weight gain and physical performance of the animals. Our results show that B2R-/- mice are resistant to high fat diet-induced obesity, have higher glucose tolerance as well as increased mitochondrial mass. These features are accompanied by higher energy expenditure and a lower feed efficiency associated with an increase in the proportion of type I fibers and intermediary fibers characterized by higher mitochondrial content and increased expression of genes related to oxidative metabolism. Additionally, the increased percentage of oxidative skeletal muscle fibers and mitochondrial apparatus in B2R-/- mice is coupled with a higher aerobic exercise performance. Taken together, our data give support to the involvement of kinins in skeletal muscle fiber type distribution and muscle metabolism, which ultimately protects against fat-induced obesity and improves aerobic exercise performance.

Erythrocytic nuclear alterations have been considered as an indicative of organism's exposure to genotoxic agents. Due to their close relationship among their frequencies and DNA damages, they are considered excellent markers of exposure in eukaryotes. However, poor data has been found in literature concerning their genesis, differential occurrence and their life span. In this study, we use markers of cell viability; genotoxicity and cellular turn over in order to shed light to these events. Tilapia and their blood were exposed to cadmium in acute exposure and in vitro assays. They were analyzed using flow cytometry for oxidative stress and membrane disruption, optical microscopy for erythrocytic nuclear alteration, graphite furnace atomic absorption spectrometry for cadmium content in aquaria water, blood and cytochemical and analytical electron microscopy techniques for the hemocateretic aspects. The results showed a close relationship among the total nuclear alterations and cadmium content in the total blood and melanomacrophage centres area, mismatching reactive oxygen species and membrane damages. Moreover, nuclear alterations frequencies (vacuolated, condensed and blebbed) showed to be associated to cadmium exposure whereas others (lobed and bud) were associated to depuration period. Decrease on nuclear alterations frequencies was also associated with hemosiderin increase inside spleen and head kidney macrophages mainly during depurative processes. These data disclosure in temporal fashion the main processes that drive the nuclear alterations frequencies and their relationship with some cellular and systemic biomarkers.

The host acceptance of neonate Alabama argillacea (Hübner) (Lepidoptera: Noctuidae) larvae to Bt cotton plants exerts a strong influence on the potential risk that this pest will develop resistance to Bt cotton. This will also determine the efficiency of management strategies to prevent its resistance such as the "refuge-in-the-bag" strategy. In this study, we assessed the acceptance of neonate A. argillacea larvae to Bt and non-Bt cotton plants at different temperatures during the first 24 h after hatching. Two cotton cultivars were used in the study, one a Bt DP 404 BG (Bollgard) cultivar, and the other, an untransformed isoline, DP 4049 cultivar. There was a greater acceptance by live neonate A. argillacea larvae for the non-Bt cotton plants compared with the Bt cotton plants, especially in the time interval between 18 and 24 h. The percentages of neonate A. argillacea larvae found on Bt or non-Bt plants were lower when exposed to temperatures of 31 and 34 °C. The low acceptance of A. argillacea larvae for Bt cotton plants at high temperatures stimulated the dispersion of A. argillacea larvae. Our results support the hypothesis that the dispersion and/or feeding behavior of neonate A. argillacea larvae is different between Bt and non-Bt cotton. The presence of the Cry1Ac toxin in Bt cotton plants, and its probable detection by the A. argillacea larvae tasting or eating it, increases the probability of dispersion from the plant where the larvae began. These findings may help to understand how the A. argillacea larvae detect the Cry1Ac toxin in Bt cotton and how the toxin affects the dispersion behavior of the larvae over time. Therefore, our results are extremely important for the management of resistance in populations of A. argillacea on Bt cotton.

The relationship between the insect development rate and temperature was established very early and represents an important ecological variable for modeling the population dynamics of insects. The accurate determination of thermal constant values and the lower and upper developmental thresholds of Hyadaphis foeniculi (Passerini) (Hemiptera: Aphididae) on fennel (Foeniculum vulgare Miller (Apiales: Apiaceae)) crops would obviously benefit the effective application of control measures. This paper is a study of the biology and thermal requirements of H. foeniculi. Winged insects were collected from fennel crops at the Embrapa Algodão in Campina Grande, Paraíba. Nymphs (age ≤24 h) produced by winged insects were subjected to constant temperatures of 15, 20, 25, 28, 30 or 33°C, a photophase of 12 h and a relative humidity of 70±10%. The results of the study showed that at temperatures between 15 and 30°C, H. foeniculi nymphs were able to develop normally. The four instars were found at all temperatures tested. However, temperatures of 3 and 33°C were lethal to the nymphs. The nymph stage development time varied from 5 (30°C) to 19 (15°C) days. The influence of temperature on the development time is dependent on the instar. The base temperature (Tb) and the thermal constant (K) for the nymph stage were estimated at 11.2°C and 107.5 degree-days, respectively. The shortest nymph development stage was observed at 30°C, and the highest nymph viability (85.0%) was observed at 28°C. This information can be used for developing phenological models based on the temperature and development rate relationships so that outbreaks of H. foeniculi in the fennel crop can be predicted, therefore improving the application of control programs targeting this fennel pest.

Malaria affects millions of people worldwide and hundreds of thousands of people each year in Brazil. The mosquito Anopheles aquasalis is an important vector of Plasmodium vivax, the main human malaria parasite in the Americas. Reactive oxygen species (ROS) have been shown to have a role in insect innate immune responses as a potent pathogen-killing agent. We investigated the mechanisms of free radicals modulation after A. aquasalis infection with P. vivax. ROS metabolism was evaluated in the vector by studying expression and activity of three key detoxification enzymes, one catalase and two superoxide dismutases (SOD3A and SOD3B). Also, the involvement of free radicals in the mosquito immunity was measured by silencing the catalase gene followed by infection of A. aquasalis with P. vivax. Catalase, SOD3A and SOD3B expression in whole A. aquasalis were at the same levels of controls at 24 h and upregulated 36 h after ingestion of blood containing P. vivax. However, in the insect isolated midgut, the mRNA for these enzymes was not regulated by P. vivax infection, while catalase activity was reduced 24 h after the infectious meal. RNAi-mediated silencing of catalase reduced enzyme activity in the midgut, resulted in increased P. vivax infection and prevalence, and decreased bacterial load in the mosquito midgut. Our findings suggest that the interactions between A. aquasalis and P. vivax do not follow the model of ROS-induced parasite killing. It appears that P. vivax manipulates the mosquito detoxification system in order to allow its own development. This can be an indirect effect of fewer competitive bacteria present in the mosquito midgut caused by the increase of ROS after catalase silencing. These findings provide novel information on unique aspects of the main malaria parasite in the Americas interaction with one of its natural vectors.

Uno de los problemas más frecuentes observados en muestras biológicas al microscopio de rastreo (SEM) es la disminución de tamaño de los especímenes, lo cual se ha asociado con el método de secado. En el caso de secado en punto crítico (SPC), esta reducción se calcula entre 20 y 80% pero, para el secado en terbutanol (STB) aún no se ha calculado. Se estudió el tamaño de las levaduras de Candida albicans y Phaeoannelomyces werneckii y de eritrocitos humanos procesados para SEM mediante SPC y STB, y contraste de fases. Las células se lavaron tres veces con amortiguador de fosfatos y se fijaron con glutaraldehído al 2,5% en el mismo amortiguador a 4°C por 2h. Se procesaron por duplicado sobre membranas policatiónicas de poli-L-lisina, se deshidrataron y se secaron en SPC y STB; se cubrieron con 200 A de oro y se analizaron al SEM (Hitachi S-570). El tamaño real se obtuvo en el microscopio de contraste de fases. Los resultados mostraron que, a pesar de las variaciones rnorfológicas presentes en las células estudiadas, hubo una disminución del 34,4 y 21,3% aproximadamente en el tamaño de los eritrocitos procesados por SPC y STB, respectivamente. Se discuten las variaciones en el tamaño del espécimen durante el procesamiento para SEM.

Se describen las características morfológicas de esporulación al microscopio electrónico de rastreo (MER) de algunos de los principales hongos encontrados en la atmósfera de Santafé de Bogotá. Los hongos se fijaron con vapores de acroleína pura y cristales de OSO, y se secaron en alto vacío, que evita el daño de las estructuras aéreas y arreglos característicos, como lo hemos informado previamente. El análisis al MER reveló la variedad morfológica de los hongos estudiados y permitió observar con detalle las características ultraestructurales de la superficie de los hongos, lo cual es de importancia en la realización del estudio morfológico, estructural y taxonómico; reveló estructuras y características morfológicas y detalles de la conidiogénesis que ayudan a un mejor entendimiento del proceso; algunos de estos detalles estructurales son difíciles de observar al microscopio de luz o al microscopio de contraste de fase.

Se evaluaron cuatro métodos de procesamiento para el estudio ultra estructural de hongos al microscopio electrónico de rastreo (MER). Se utilizaron microcultivos de cepas colombianas de Fonsecaea pedrosoi aisladas de lesiones de pacientes en el Instituto Dermatológico Federico Lleras Acosta, Santafé de Bogotá. Los resultados mostraron alteraciones, distorsiones o artificios en los hongos procesados por tres de los métodos utilizados, debido probablemente a los efectos físicos o químicos durante el procesamiento. Se propone el método de fijación con vapores de acroleína pura y cristales de OSO. con deshidratación y secado en alto vacío que evitó la pérdida de estructuras y conservo los arreglos característicos, presentando rapidez, fácil manipulación del espécimen y disminución de reactivos. Se destaca la importancia del MER en el estudio morfológico, estructural y taxonómico de los hongos y el uso de los vapores de acroleína para la fijación y el alto vacío para el secado, como alternativas metodológicas para el procesamiento de especímenes biológicos de difícil manipulación o que presentan fácil alteración por su composición higrófila.

BackgroundIn experimental models, hypothalamic inflammation is an early and determining factor in the installation and progression of obesity. Pharmacological and gene-based approaches have proven efficient in restraining inflammation and correcting the obese phenotypes. However, the role of nutrients in the modulation of hypothalamic inflammation is unknown.Methodology/principal findingsHere we show that, in a mouse model of diet-induced obesity, partial substitution of the fatty acid component of the diet by flax seed oil (rich in C18:3) or olive oil (rich in C18:1) corrects hypothalamic inflammation, hypothalamic and whole body insulin resistance, and body adiposity. In addition, upon icv injection in obese rats, both ω3 and ω9 pure fatty acids reduce spontaneous food intake and body mass gain. These effects are accompanied by the reversal of functional and molecular hypothalamic resistance to leptin/insulin and increased POMC and CART expressions. In addition, both, ω3 and ω9 fatty acids inhibit the AMPK/ACC pathway and increase CPT1 and SCD1 expression in the hypothalamus. Finally, acute hypothalamic injection of ω3 and ω9 fatty acids activate signal transduction through the recently identified GPR120 unsaturated fatty acid receptor.Conclusions/significanceUnsaturated fatty acids can act either as nutrients or directly in the hypothalamus, reverting diet-induced inflammation and reducing body adiposity. These data show that, in addition to pharmacological and genetic approaches, nutrients can also be attractive candidates for controlling hypothalamic inflammation in obesity.

Wound healing is impaired in diabetes mellitus, but the mechanisms involved in this process are virtually unknown. Proteins belonging to the insulin signaling pathway respond to insulin in the skin of rats.The purpose of this study was to investigate the regulation of the insulin signaling pathway in wound healing and skin repair of normal and diabetic rats, and, in parallel, the effect of a topical insulin cream on wound healing and on the activation of this pathway.We investigated insulin signaling by immunoblotting during wound healing of control and diabetic animals with or without topical insulin. Diabetic patients with ulcers were randomized to receive topical insulin or placebo in a prospective, double-blind and placebo-controlled, randomized clinical trial (NCT 01295177) of wound healing.Expression of IR, IRS-1, IRS-2, SHC, ERK, and AKT are increased in the tissue of healing wounds compared to intact skin, suggesting that the insulin signaling pathway may have an important role in this process. These pathways were attenuated in the wounded skin of diabetic rats, in parallel with an increase in the time of complete wound healing. Upon topical application of insulin cream, the wound healing time of diabetic animals was normalized, followed by a reversal of defective insulin signal transduction. In addition, the treatment also increased expression of other proteins, such as eNOS (also in bone marrow), VEGF, and SDF-1α in wounded skin. In diabetic patients, topical insulin cream markedly improved wound healing, representing an attractive and cost-free method for treating this devastating complication of diabetes.ClinicalTrials.gov NCT01295177.

BACKGROUND: Buruli ulcer (BU) is a neglected necrotizing disease of the skin, subcutaneous tissue and bone, caused by Mycobacterium ulcerans. BU pathogenesis is associated with mycolactone, a lipidic exotoxin with cytotoxic and immunosuppressive properties. Since 2004, the World Health Organization recommends the treatment of BU with a combination of rifampicin and streptomycin (RS). Histological analysis of human tissue samples suggests that such antibiotic treatment reverses the mycolactone-induced local immunosuppression, leading to increased inflammatory infiltrations and phagocytosis of bacilli. METHODOLOGY/PRINCIPAL FINDINGS: We used a mouse model of M. ulcerans footpad infection, followed by combined RS treatment. Time-lapsed analyses of macroscopic lesions, bacterial burdens, histology and immunohistochemistry were performed in footpads. We also performed CFU counts, histology and immunohistochemistry in the popliteal draining lymph nodes (DLN). We observed a shift in the cellular infiltrates from a predominantly neutrophilic/macrophagic to a lymphocytic/macrophagic profile in the infected footpads of antibiotic-treated mice. This shift occurred before the elimination of viable M. ulcerans organisms, which were ultimately eradicated as demonstrated by the administration of dexamethasone. This reduction of bacillary loads was accompanied by an increased expression of inducible nitric oxide synthase (NOS2 or iNOS). Predominantly mononuclear infiltrates persisted in the footpads during and after treatment, coincident with the long persistence of non-viable poorly stained acid-fast bacilli (AFB). We additionally observed that antibiotherapy prevented DLN destruction and lymphocyte depletion, which occurs during untreated experimental infections. CONCLUSIONS/SIGNIFICANCE: Early RS treatment of M. ulcerans mouse footpad infections results in the rapid elimination of viable bacilli with pathogen eradication. However, non-viable AFB persisted for several months after lesion sterilization. This RS regimen prevented DLN destruction, allowing the rapid re-establishment of local and regional cell mediated immune responses associated with macrophage activation. Therefore it is likely that this re-establishment of protective cellular immunity synergizes with antibiotherapy.

BackgroundHeme-copper oxygen reductases (HCOs) are the last enzymatic complexes of most aerobic respiratory chains, reducing dioxygen to water and translocating up to four protons across the inner mitochondrial membrane (eukaryotes) or cytoplasmatic membrane (prokaryotes). The number of completely sequenced genomes is expanding exponentially, and concomitantly, the number and taxonomic distribution of HCO sequences. These enzymes were initially classified into three different types being this classification recently challenged.MethodologyWe reanalyzed the classification scheme and developed a new bioinformatics classifier for the HCO and Nitric oxide reductases (NOR), which we benchmark against a manually derived gold standard sequence set. It is able to classify any given sequence of subunit I from HCO and NOR with a global recall and precision both of 99.8%. We use this tool to classify this protein family in 552 completely sequenced genomes.ConclusionsWe concluded that the new and broader data set supports three functional and evolutionary groups of HCOs. Homology between NORs and HCOs is shown and NORs closest relationship with C Type HCOs demonstrated. We established and made available a classification web tool and an integrated Heme-Copper Oxygen reductase and NOR protein database (www.evocell.org/hco).

The cellular form of the prion protein, PrP(C), undergoes extensive proteolysis at the alpha site (109K [see text]H110). Expression of non-cleavable PrP(C) mutants in transgenic mice correlates with neurotoxicity, suggesting that alpha-cleavage is important for PrP(C) physiology. To gain insights into the mechanisms of alpha-cleavage, we generated a library of PrP(C) mutants with mutations in the region neighbouring the alpha-cleavage site. The prevalence of C1, the carboxy adduct of alpha-cleavage, was determined for each mutant. In cell lines of disparate origin, C1 prevalence was unaffected by variations in charge and hydrophobicity of the region neighbouring the alpha-cleavage site, and by substitutions of the residues in the palindrome that flanks this site. Instead, alpha-cleavage was size-dependently impaired by deletions within the domain 106-119. Almost no cleavage was observed upon full deletion of this domain. These results suggest that alpha-cleavage is executed by an alpha-PrPase whose activity, despite surprisingly limited sequence specificity, is dependent on the size of the central region of PrP(C).

BACKGROUND:Several effects of leptin in the immune system rely on its capacity to modulate cytokine expression and apoptosis in the thymus. Surprisingly, some of these effects are dependent on signal transduction through the IRS1/PI3-kinase, but not on the activation of JAK2. Since all the well known effects of leptin in different cell types and tissues seem to be dependent on JAK2 activation, we hypothesized that, at least for the control of thymic function, another, unknown kinase could mediate the transduction of the leptin signal from the ObR towards the IRS1/PI3-kinase signaling cascade. METHODOLOGY/PRINCIPAL FINDINGS:Here, by employing immunoblot, real-time PCR and flow citometry we show that the tyrosine kinase, Fyn, is constitutively associated with the ObR in thymic cells. Following a leptin stimulus, Fyn undergoes an activating tyrosine phosphorylation and a transient association with IRS1. All these effects are independent of JAK2 activation and, upon Fyn inhibition, the signal transduction towards IRS1/PI3-kinase is abolished. In addition, the inhibition of Fyn significantly modifies the effects of leptin on thymic cytokine expression. CONCLUSION/SIGNIFICANCE:Therefore, in the thymus, Fyn acts as a tyrosine kinase that transduces the leptin signal independently of JAK2 activation, and mediates some of the immunomodulatory effects of leptin in this tissue.

Consumption of dietary fats is amongst the most important environmental factors leading to obesity. In rodents, the consumption of fat-rich diets blunts leptin and insulin anorexigenic signaling in the hypothalamus by a mechanism dependent on the in situ activation of inflammation. Since inflammatory signal transduction can lead to the activation of apoptotic signaling pathways, we evaluated the effect of high-fat feeding on the induction of apoptosis of hypothalamic cells. Here, we show that consumption of dietary fats induce apoptosis of neurons and a reduction of synaptic inputs in the arcuate nucleus and lateral hypothalamus. This effect is dependent upon diet composition, and not on caloric intake, since pair-feeding is not sufficient to reduce the expression of apoptotic markers. The presence of an intact TLR4 receptor, protects cells from further apoptotic signals. In diet-induced inflammation of the hypothalamus, TLR4 exerts a dual function, on one side activating pro-inflammatory pathways that play a central role in the development of resistance to leptin and insulin, and on the other side restraining further damage by controlling the apoptotic activity.

AMP-activated protein kinase (AMPK) and mammalian Target of Rapamycin (mTOR) are key regulators of cellular energy balance and of the effects of leptin on food intake. Acute exercise is associated with increased sensitivity to the effects of leptin on food intake in an IL-6-dependent manner. To determine whether exercise ameliorates the AMPK and mTOR response to leptin in the hypothalamus in an IL-6-dependent manner, rats performed two 3-h exercise bouts, separated by one 45-min rest period. Intracerebroventricular IL-6 infusion reduced food intake and pretreatment with AMPK activators and mTOR inhibitor prevented IL-6-induced anorexia. Activators of AMPK and fasting increased food intake in control rats to a greater extent than that observed in exercised ones, whereas inhibitor of AMPK had the opposite effect. Furthermore, the reduction of AMPK and ACC phosphorylation and increase in phosphorylation of proteins involved in mTOR signal transduction, observed in the hypothalamus after leptin infusion, were more pronounced in both lean and diet-induced obesity rats after acute exercise. Treatment with leptin reduced food intake in exercised rats that were pretreated with vehicle, although no increase in responsiveness to leptin-induced anorexia after pretreatment with anti-IL6 antibody, AICAR or Rapamycin was detected. Thus, the effects of leptin on the AMPK/mTOR pathway, potentiated by acute exercise, may contribute to appetite suppressive actions in the hypothalamus.

Fetal hydrothorax refers to a collection of fluid within the fetal thorax that may be the result of chylous leak from the thoracic duct (primary hydrothorax) or generalized fluid retention associated with immune or no immune fetal hydrops (secondary hydrothorax). The authors' presents a case report occurred in 2002, of a pregnant woman that at 25 weeks' gestation that was referred to Maternidade Bissaya-Barreto-Coimbra because of a fetal hydrothorax at left, under tension and with cardiac decompensation signs. A fetal thoracocentesis was performed and the diagnosis was chylothorax. Because of a rapid reaccumulation of fluid a pleuroamniotic shunt was placed. The effusion and the cardiac decompensation signs regressed. The delivery was at 38 weeks' gestation. The newborn had been stable. Actually he has 10 months, is healthy and has a normal grow and development.

Os fundamentos da semiologia ortopédica são apresentados, divididos em: inspeção geral, exame da marcha o dos segmentos do sistema músculo-esquelético: coluna vertebral, ombros, cotovelos, mãos, quadris, joelhos e pés.

Abstract Inferior frontal sulcal hyperintensities (IFSH) observed on fluid-attenuated inversion recovery (FLAIR) MRI have been proposed as indicators of elevated cerebrospinal fluid waste accumulation in cerebral small vessel disease (CSVD). However, to validate IFSH as a reliable imaging biomarker, further replication studies are required. The objective of this study was to investigate associations between IFSH and CSVD, and their potential repercussions, i.e., cognitive impairment and depression. We prospectively recruited 47 patients with CSVD and 29 cognitively normal controls (NC). IFSH were rated visually based on FLAIR MRI. Using different regression models, we explored the relationship between IFSH, group status (CSVD vs. NC), CSVD severity assessed with MRI, cognitive function, and symptoms of depression. Patients with CSVD were more likely to have higher IFSH scores compared to NC (OR 5.64, 95% CI 1.91–16.60), and greater CSVD severity on MRI predicted more severe IFSH (OR 1.47, 95% CI 1.14–1.88). Higher IFSH scores were associated with lower cognitive function (-0.96, 95% CI -1.81 to -0.10), and higher levels of depression (0.33, 95% CI 0.01–0.65). CSVD and IFSH may be tightly linked to each other, and the accumulation of waste products, indicated by IFSH, could have detrimental effects on cognitive function and symptoms of depression.

Abstract Cognition plays a central role in the diagnosis and characterization of dementia with Lewy bodies (DLB). However, the complex associations among cognitive deficits in different domains in DLB are largely unknown. To characterize these associations, we investigated and compared the cognitive connectome of DLB patients, healthy controls (HC), and Alzheimer’s disease patients (AD). We obtained data from the National Alzheimer’s Coordinating Center. We built cognitive connectomes for DLB (n = 104), HC (n = 3703), and AD (n = 1985) using correlations among 24 cognitive measures mapping multiple cognitive domains. Connectomes were compared using global and nodal graph measures of centrality, integration, and segregation. For global measures, DLB showed a higher global efficiency (integration) and lower transitivity (segregation) than HC and AD. For nodal measures, DLB showed higher global efficiency in most measures, higher participation (centrality) in free-recall memory, processing speed/attention, and executive measures, and lower local efficiency (segregation) than HC. Compared with AD, DLB showed lower nodal strength and local efficiency, especially in memory consolidation. The cognitive connectome of DLB shows a loss of segregation, leading to a loss of cognitive specialization. This study provides the data to advance the understanding of cognitive impairment and clinical phenotype in DLB, with implications for differential diagnosis.

Abstract Objective The inflammatory responses from synovial fibroblasts and macrophages and the mitochondrial dysfunction in chondrocytes lead to oxidative stress, disrupt extracellular matrix (ECM) homeostasis, and accelerate the deterioration process of articular cartilage in osteoarthritis (OA). In recent years, it has been proposed that mesenchymal stromal cells (MSC) transfer their functional mitochondria to damaged cells in response to cellular stress, becoming one of the mechanisms underpinning their therapeutic effects. Therefore, we hypothesize that a novel cell-free treatment for OA could involve direct mitochondria transplantation, restoring both cellular and mitochondrial homeostasis. Methods Mitochondria were isolated from Umbilical Cord (UC)-MSC (Mito-MSC) and characterized based on their morphology, phenotype, functions, and their ability to be internalized by different articular cells. Furthermore, the transcriptional changes following mitochondrial uptake by chondrocytes were evaluated using an Affymetrix analysis, Lastly, the dose dependence therapeutic efficacy, biodistribution and immunogenicity of Mito-MSC were assessed in vivo, through an intra-articular injection in male C57BL6 mice in a collagenase-induced OA (CIOA) model. Results Our findings demonstrate the functional integrity of Mito-MSC and their ability to be efficiently transferred into chondrocytes, synovial macrophages, and synovial fibroblasts. Moreover, the transcriptomic analysis showed the upregulation of genes involved in stress such as DNA reparative machinery and inflammatory antiviral responses. Finally, Mito-MSC transplantation yielded significant reductions in joint mineralization, a hallmark of OA progression, as well as improvements in OA-related histological signs, with the lower dose exhibiting better therapeutic efficacy. Furthermore, Mito-MSC was detected within the knee joint for up to 24 h post-injection without eliciting an inflammatory response in CIOA mice. Conclusion Collectively, our results reveal that mitochondria derived from MSC are transferred to key articular cells and are retained in the joint without generating an inflammatory immune response mitigating articular cartilage degradation in OA, probably through a restorative effect triggered by the stress antiviral response within OA chondrocytes.

Abstract Body composition abnormalities are prognostic markers in several types of cancer, including colorectal cancer (CRC). Using our data distribution on body composition assessments and classifications could improve clinical evaluations and support population-specific opportune interventions. This study aimed to evaluate the distribution of body composition from computed tomography and assess the associations with overall survival among patients with CRC. In this multicenter cohort study, patients (N = 635) aged 18 years and older with CRC were observed for 12 to 36 months to assess outcomes. Skeletal muscle area (SM) and index (SMI), skeletal muscle radiodensity (SMD), intermuscular adipose tissue (IMAT), subcutaneous adipose tissue (SAT), and visceral adipose tissue (VAT) were evaluated, and classified based on tertile distributions. Low muscle mass (SMI) and poor muscle composition (SMD) were independent predictors of mortality regardless of follow-up period. This risk of mortality increased to more than 3-fold when combining both low SMI and low SMD (HRadjusted 3.1, 95% CI 1.8 to 5.4, respectively). Our study indicates that body composition characteristics may vary across countries, highlighting the need for developing sex- and population-specific cutoff values for computed tomography assessments in patients with different types of cancer.

Abstract The potential of the island of La Palma (Canary Islands) to host geothermal resources is very high, mainly due to its high volcanic activity. The primary goal of this study is to get a tridimensional image of the seismic intrinsic attenuation using ambient seismic noise and to identify anomalies that may be linked to active geothermal reservoirs on La Palma island. For this purpose, we developed a new Ambient Noise Attenuation Tomography (ANAT) technique, which uses seismic ambient noise for imaging intrinsic attenuation in 3-D at a local scale down to 5 km depth. Our research identifies two areas with high attenuation in the island’s southern region. One area could be associated with hydrothermal alteration zones beneath the Cumbre Vieja volcanic complex. Another high-attenuation zone was observed in the island’s southern part, which could be associated with extensively fractured rocks that might facilitate the circulation of heated fluids. Furthermore, we discuss the geothermal relevance of such anomalies, making a comparison with previous resistivity, S-wave velocity and density models. This study confirms that intrinsic attenuation retrieved from the coda of Rayleigh waves is more sensitive to the presence of fluids than velocity. Fluids being a key component of active geothermal reservoirs, it is reasonable to expect intrinsic attenuation anomalies in these systems. Therefore, we conclude that ANAT can be useful for geothermal exploration.