In Situ/Subcellular Localization of Arabinogalactan Protein Expression by Fluorescent In Situ Hybridization (FISH)

The arabinogalactan proteins are highly glycosylated and ubiquitous in plants. They are involved in several aspects of plant development and reproduction; however, the mechanics behind their function remains for the most part unclear, as the carbohydrate moiety, covering the most part of the protein core, is poorly characterized at the individual protein level. Traditional immunolocalization using antibodies that recognize the glycosidic moiety of the protein cannot be used to elucidate individual proteins' distribution, function, or interactors. Indirect approaches are typically used to study these proteins, relying on reverse genetic analysis of null mutants or using a reporter fusion system. In the method presented here, we propose the use of RNA probes to assist in the localization of individual AGPs expression/mRNAs in tissues of Arabidopsis by fluorescent in situ hybridization, FISH. An extensive description of all aspects of this technique is provided, from RNA probe synthesis to the hybridization, trying to overcome the lack of specific antibodies for the protein core of AGPs

Single Fetal Death in Twin Pregnancy. What Will Be the Problems to the Surviving Co-Sib?

Na gravidez bigemelar, a morte de um dos fetos no segundo e terceiro trimestre é uma complicação rara, variando a taxa de incidência entre 2,6 e 6,8%. Este acontecimento determina um aumento das taxas de morbilidade e mortalidade perinatal para o gémeo sobrevivente, especialmente em gravidezes monocoriónicas, quando a morte fetal é devida à síndroma de tranfusão feto-fetal. Para alguns autores o atraso de crescimento intrauterino e a prematuridade são os principais factores de risco para o aumento da morbilidade e mortalidade do gémeo sobrevivente. A patofisiologia de instalação dos distúrbios cerebrais no gémeo sobrevivente após a morte do feto irmão não está definida, nem o intervalo de tempo que medeia entre a morte e o estabelecimento das lesões. Também não existe um protocolo definitivo de seguimento destas gravidezes e, posteriormente, do gémeo sobrevivente. No período de 1 de Setembro de 1994 a 31 de Dezembro de 1998, foram seguidas, na consulta de Gravidez Múltipla da Maternidade Dr. Alfredo da Costa, 235 gravidezes bigemelares. Em nove casos (3,8%) ocorreu morte de um dos fetos com idade gestacional acima das 13 semanas. Em cinco das nove gravidezes foi conhecida a causa de morte, quatro das quais foram atribuídas à síndroma de tranfusão feto-fetal. A taxa de prematuridade do gémeo sobrevivente foi de 44,4% (4/9) e a de mortalidade de 11,1%(1/9). A taxa de morbilidade neonatal foi de 62,5% (5/8), na maioria dos casos por complicações inerentes à prematuridade. A taxa de morbilidade neurológica foi de 37,5% (3/8). A taxa de lesões neurológicas major foi de 25% (2/8) e ocorreu em recém-nascidos de termo. A síndroma de transfusão feto-fetal, como causa de morte fetal, associou-se aos casos com pior prognóstico no que se referiu ao gémeo sobrevivente.O crescimento do gémeo sobrevivente parece depender das lesões provocadas pela morte do feto irmão. Os autores finalizam com uma proposta de atitudes obstétricas e pediátricas em relação ao gémeo sobrevivente

A New Relativistic Component of the Accretion Disk Wind in PDS 456

Past X-ray observations of the nearby luminous quasar PDS 456 (at z = 0.184) have revealed a wide angle accretion disk wind, with an outflow velocity of ∼−0.25 c . Here, we unveil a new, relativistic component of the wind through hard X-ray observations with NuSTAR and XMM-Newton , obtained in 2017 March when the quasar was in a low-flux state. This very fast wind component, with an outflow velocity of −0.46 ± 0.02 c , is detected in the iron K band, in addition to the −0.25 c wind zone. The relativistic component may arise from the innermost disk wind, launched from close to the black hole at a radius of ∼10 gravitational radii. The opacity of the fast wind also increases during a possible obscuration event lasting for 50 ks. We suggest that the very fast wind may only be apparent during the lowest X-ray flux states of PDS 456, becoming overly ionized as the luminosity increases. Overall, the total wind power may even approach the Eddington value

Peripheral Blood Leukocytes And Serum Nested Polymerase Chain Reaction Are Complementary Methods For Monitoring Active Cytomegalovirus Infection In Transplant Patients.

Human cytomegalovirus is an important cause of morbidity and mortality in immunocompromised patients. Qualitative polymerase chain reaction (PCR) has proven to be a sensitive and effective technique in defining active cytomegalovirus infection, in addition to having low cost and being a useful test for situations in which there is no need for quantification. Real-time PCR has the advantage of quantification; however, the high cost of this methodology makes it impractical for routine use. To apply a nested PCR assay to serum (sPCR) and to evaluate its efficiency to diagnose active cytomegalovirus infection compared with PCR of peripheral blood leukocytes (L-PCR). Samples of 37 patients were prospectively evaluated. An internal control was created and applied to sPCR to exclude false-negative results. In total, 21 patients (57%) developed active cytomegalovirus infection. After analyzing the two methods for the diagnosis of active infection, higher sensitivity and negative predictive value of the L-PCR versus sPCR (100% versus 62%), and higher specificity and positive predictive value of sPCR versus L-PCR (81% versus 50% and 72%, respectively) were observed. Discordant results were observed in 11 patients who were L-PCR-positive but sPCR-negative for active cytomegalovirus infection, five of whom developed clinical symptoms of cytomegalovirus. Clinical symptoms were observed in 14 patients, 12 of whom were diagnosed with active infection by nested L-PCR (P=0.007) and seven by nested sPCR (P=0.02). Higher specificity and a positive predictive value for sPCR were observed. Nested L-PCR and sPCR were considered to be complementary methods for the diagnosis and management of symptomatic cytomegalovirus infection.24e69-7

Effect of physical exercise on lipid metabolism in the elderly

PURPOSE: To assess the effect of regular physical exercise on plasma lipids and apoproteins in an elderly group. MATERIAL AND METHODS: Sixty-three old people of both sexes, aged between 65 and 94, were randomly distributed between a test group (n = 31) and a control group (n = 32). The test group followed a program of physical exercise for eight months, with intensity of 60% to 80% of heart rate reserve, consisting of three weekly sessions, on alternate days, of 60 minutes each. Before the beginning of the program and eight months afterwards, blood samples were collected to assess total cholesterol, triglycerides, HDL-C, HDL2-C. HDL3-C, apo A1, apo B-100 and apo (a). The ratios total cholesterol/HDL-C, LDL-C/HDL-C, HDL2-C/HDL3-C and apo A1/apo B-100 were also calculated. RESULTS: In the test group total cholesterol, triglycerides, LDL-C, apo B-100, total cholesterol/HDL-C and LDL-C/HDL-C decreased significantly; HDL-C, HDL2-C, apo A1, HDL2-C/HDL3-C and apo A1/apo B-100 increased significantly; HDL3-C and apo (a) did not change. The control group did not present any significant alterations in any of the parameters assessed. CONCLUSIONS: Regular physical exercise decreases vascular risk and may help to reduce cardiovascular events in the elderly

Evidence for a radiatively driven disc-wind in PDS 456?

We present a newly discovered correlation between the wind outflow velocity and the X-ray luminosity in the luminous ($L_{\rm bol}\sim10^{47}\,\rm erg\,s^{-1}$) nearby ($z=0.184$) quasar PDS\,456. All the contemporary XMM-Newton, NuSTAR and Suzaku observations from 2001--2014 were revisited and we find that the centroid energy of the blueshifted Fe\,K absorption profile increases with luminosity. This translates into a correlation between the wind outflow velocity and the hard X-ray luminosity (between 7--30\,keV) where we find that $v_{\rm w}/c \propto L_{7-30}^{\gamma}$ where $\gamma=0.22\pm0.04$. We also show that this is consistent with a wind that is predominately radiatively driven, possibly resulting from the high Eddington ratio of PDS\,456

Linking forest cover, soil erosion and mire hydrology to late-Holocene human activity and climate in NW Spain

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 3.0 License (http://www.creativecommons.org/licenses/by-nc/3.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (http://www.uk.sagepub.com/aboutus/openaccess.htm).This article has been made available through the Brunel Open Access Publishing Fund.Forest clearance is one of the main drivers of soil erosion and hydrological changes in mires, although climate may also play a significant role. Because of the wide range of factors involved, understanding these complex links requires long-term multi-proxy approaches and research on the best proxies to focus. A peat core from NW Spain (Cruz do Bocelo mire), spanning the last ~3000 years, has been studied at high resolution by physical (density and loss on ignition (LOI)), geochemical (elemental composition) and palynological (pollen and non-pollen palynomorphs) analyses. Proxies related to mineral matter fluxes from the catchment (lithogenic tracers, Glomus and Entorrhiza), rainfall (Bromine), mire hydrology (HdV-18), human pressure (Cerealia-type, nitrophilous taxa and coprophilous fungi) and forest cover (mesophilous tree taxa) were the most useful to reconstruct the evolution of the mire and its catchment. Forest clearance for farming was one of the main drivers of environmental change from at least the local Iron Age (~2685 cal. yr BP) onwards. The most intense phase of deforestation occurred during Roman and Germanic times and the late Middle Ages. During these phases, the entire catchment was affected, resulting in enhanced soil erosion and severe hydrological modifications of the mire. Climate, especially rainfall, may have also accelerated these processes during wetter periods. However, it is noteworthy that the hydrology of the mire seems to have been insensitive to rainfall variations when mesophilous forest dominated. Abrupt changes were only detected once intense forest clearance commenced during the Iron Age/Roman transition (~2190 cal. yr BP) phase, which represented a tipping point in catchment's ability to buffer impacts. Overall, our findings highlight the importance of studying ecosystems' long-term trajectories and catchment-wide processes when implementing mire habitat protection measures.This work was funded by the projects CGL2010-20672 (Plan Nacional I+D+i, Spanish Ministry of Science and Innovation) and 10PXIB200182PR (General Directorate of I+D, Xunta de Galicia). N Silva-Sánchez and L López-Merino are currently supported by a FPU predoctoral scholarship (AP2010-3264) funded by the Spanish Government and a MINT postdoctoral fellowship funded by the Brunel Institute for the Environment, respectively

Inflammation in Bipolar Disorder: Identification of New Therapeutic Targets

Introdução: A doença bipolar é um distúrbio mental com origem no conceito “mania”. Trata-se de uma patologia crónica, de natureza cíclica, caracterizada pelo aparecimento de episódios alternados de mania/hipomania com episódios de eutimia/depressão. Com este artigo pretendemos identificar novas abordagens terapêuticas para o tratamento da doença bipolar, tendo em consideração o envolvimento da inflamação na sua fisiopatologia. Métodos: Revisão da literatura que relaciona a doença bipolar com a inflamação e possíveis alvos terapêuticos, baseada em artigos de revisão. Pesquisa de ensaios clínicos que permitam avaliar as novas intervenções terapêuticas. Resultados: O tratamento farmacológico da doença bipolar é complexo, sendo periodicamente adaptado à fase de doença. Para a compreensão da fisiopatologia da doença bipolar são propostas várias hipóteses relacionadas com o processo inflamatório subjacente, como a ativação patológica da microglia, desvendando um vasto leque de alvos terapêuticos. Atualmente estão a decorrer alguns ensaios clínicos que avaliam a eficácia das novas estratégias terapêuticas. Conclusão: Distúrbios mentais como a doença bipolar têm suscitado preocupação na comunidade científica e médica devido ao seu impacto ao nível social e económico. Tendo em conta o envolvimento da inflamação na fisiopatologia da doença bipolar, o sistema imunitário constitui um novo alvo terapêutico. Assim, surgem novas opções de tratamento que incluem fármacos anti-inflamatórios não esteroides e antagonistas do glutamato. Estas novas abordagens requerem mais estudos, nomeadamente a realização de ensaios clínicos.info:eu-repo/semantics/publishedVersio

The Prognostic Value of Non-Linear Analysis of Heart Rate Variability in Patients with Congestive Heart Failure—A Pilot Study of Multiscale Entropy

AIMS: The influences of nonstationarity and nonlinearity on heart rate time series can be mathematically qualified or quantified by multiscale entropy (MSE). The aim of this study is to investigate the prognostic value of parameters derived from MSE in the patients with systolic heart failure. METHODS AND RESULTS: Patients with systolic heart failure were enrolled in this study. One month after clinical condition being stable, 24-hour Holter electrocardiogram was recording. MSE as well as other standard parameters of heart rate variability (HRV) and detrended fluctuation analysis (DFA) were assessed. A total of 40 heart failure patients with a mea age of 56±16 years were enrolled and followed-up for 684±441 days. There were 25 patients receiving β-blockers treatment. During follow-up period, 6 patients died or received urgent heart transplantation. The short-term exponent of DFA and the slope of MSE between scale 1 to 5 were significantly different between patients with or without β-blockers (p = 0.014 and p = 0.028). Only the area under the MSE curve for scale 6 to 20 (Area(6-20)) showed the strongest predictive power between survival (n = 34) and mortality (n = 6) groups among all the parameters. The value of Area(6-20)21.2 served as a significant predictor of mortality or heart transplant (p = 0.0014). CONCLUSION: The area under the MSE curve for scale 6 to 20 is not relevant to β-blockers and could further warrant independent risk stratification for the prognosis of CHF patients