Usability of the SedLine® electroencephalographic monitor of depth of anaesthesia in pigs: a pilot study.

To investigate the usability of the SedLine® monitor in anaesthetized pigs. Five juvenile healthy pigs underwent balanced isoflurane-based general anaesthesia for surgical placement of a subcutaneous jugular venous port. The SedLine® was applied to continuously monitor electroencephalographic (EEG) activity and its modulation during anaesthesia. Computer tomography and magnetic resonance were performed to investigate the relationship between electrodes' positioning and anatomical structures. The pediatric SedLine® EEG-sensor could be easily applied and SedLine®-generated variables collected. An EEG Density Spectral Array (DS) was displayed over the whole procedure. During surgery, the EEG signal was dominated by elevated power in the delta range (0.5-4 Hz), with an underlying broadband signal (where power decreased with increasing frequency). The emergence period was marked by a decrease in delta power, and a more evenly distributed power over the 4-40 Hz frequency range. From incision to end of surgery, mean SedLine®-generated values (± standard deviation) were overall stable [23.0 (± 2.8) Patient State Index (PSI), 1.0% (± 3.8%) Suppression Ratio (SR), 8.8 Hz (± 2.5 Hz) Spectral Edge Frequency 95% (SEF) left, 7.7 Hz (± 2.4 Hz) SEF right], quickly changing during emergence [75.3 (± 11.1) PSI, 0.0 (± 0.0) SR, 12.5 (± 6.6) SEF left 10.4 (± 6.6) SEF right]. Based on the imaging performed, the sensor does not record EEG signals from the same brain areas as in humans. SedLine®-DSA and -generated variables seemed to reflect variations in depth of anaesthesia in pigs. Further studies are needed to investigate this correlation, as well as to define the species-specific brain structures monitored by the EEG-sensor

The hidden fragility in the heart of the athletes: A review of genetic biomarkers

Sudden cardiac death (SCD) is a devastating event which can also affect people in apparent good health, such as young athletes. It is known that intense and continuous exercise along with a genetic background that predisposes a person to the risk of fatal arrhythmias is a trigger for SCD. Therefore, knowledge of the athlete’s genetic conditions underlying the onset of SCD must be extended, in order to develop new effective prevention and/or therapeutic strategies. Arrhythmic features occur across a broad spectrum of cardiac diseases, sometimes presenting with overlapping phenotypes. The genetic basis of arrhythmogenic disorders has been greatly highlighted in the last 30 years, and has shown marked heterogeneity. The advent of next-generation sequencing has constantly updated our understanding of the genetic basis of arrhythmogenic diseases and is laying the foundation for precision medicine. With the exception of a few clinical cases involving a single athlete showing a highly suspected phenotype for the presence of a heart disease, there are few studies to date that analysed the applicability of genetic testing on cohorts of athletes. This evidence shows that genetic testing can contribute to the diagnosis of up to 13% of athletes; however, the presence of clinical markers is essential. This review aims to provide a reference collection on current knowledge of the genetic basis of sudden cardiac death in athletes and to review updated evidence on the effectiveness of genetic testing in early identification of athletes at risk for SCD

Health related quality of life outcomes in HIV-Infected patients starting different combination regimens in a randomised multinational trial: the INITIO-QoL Substudy

The health-related quality of life (HRQoL) outcomes in HIV-infected, treatment-naive patients starting different HAART regimens in a 3-year, randomized, multinational trial were compared. HRQoL was measured in a subgroup of patients enrolled in the INITIO study (153/911), using a modified version of the MOS-HIV questionnaire. The regimens compared in the INITIO trial were composed by two NRTIs (didanosine + stavudine) plus either an NNRTI (efavirenz) or a PI (nelfinavir), or both (efavirenz + nelfinavir). Primary HRQoL outcomes were Physical and Mental Health Summary scores (PHS and MHS, respectively). During follow-up, an increase of PHS score was observed in all treatment arms. The MHS score remained substantially unchanged with the four-drug combination and showed with both NNRTI- and PI-based three-drug regimens a marked trend toward improvement, which became statistically significant when a multiple imputation method was used to adjust for missing data. Overall, starting all the combination regimens compared in the INITIO study was associated with a maintained or slightly improved HRQOL status, consistently with the positive immunological and virological changes observed in the main study. The observed differences in the MHS indicate a possible HRQoL benefit associated to the use of three-drug, two-class regimens and no additional benefit for the use of four-drug, three-class regimens, confirming that three-drug, two-class regimens that include two NRTIs plus either an NNRTI or a PI should be preferred as initial treatment of HIV infection

Análisis de diversidad genética en poblaciones de Bauhinia forficata subsp. pruinosa (Pezuña de vaca, o buey, Pata de vaca) mediante el uso de marcadores moleculares

Bauhinia. forficata Link. subsp. pruinosa (Vogel) Fortunato & Wunderlin, crece naturalmente desde Paraguay, Sur de Brasil hasta Argentina, y tiene antecedente de uso ornamental y terapéutico (diurético, antidiarreico, hipoglucemiante). Este proyecto tiene como objetivo la caracterización de poblaciones de bauhinia forficata subsp pruinosa utilizando marcadores moleculares. Existen SSRs diseñados en taxones afines: Cercis canadensis L. y C. chinensis, de los que hay antecedentes de transferibilidad entre especies. Por ello se evaluó su transferencia en las poblaciones de Bauhinia. Estos resultados asociados a los que se obtengan de las evaluaciones químicas permitirán conocer su relación con los biotipos que se identifiquen.  Para esto, se extrajo ADN mediante el método de Dellaporta (1983) de 10 individuos del Jardín botánico Arturo E. Ragonese, se cuantificó y se realizó PCR de los SSRs seleccionados. El producto se sembró, junto con un marcador de peso molecular, en geles desnaturalizantes de poliacrilamida en cubas de secuenciación teñidos con nitrato de plata y revelados con hidróxido de sodio. Hasta el momento, se evaluaron 15 SSRs (6 provinieron de C. chinensis   y 9 de C. canandensis). De estos 15 SSRs, 11 mostraron productos de amplificación. De ellos 9 fueron polimórficos indicando diferencias interpoblacionales, que podrían indicar variabilidad química y por lo tanto resta correlacionar la actividad biológica de los distintos biotipos. Además, algunos SSRs mostraron más de un locus, información que respalda lo señalado por Poggio et al. que en la subfamilia Cercidoideae, Bauhinia (2n=28) se generó por hibridación y poliploídia a partir del ancestro Cercis (2n= 14). Además, se realizó el muestreo de 20 puntos de colecta distribuidas en las provincias de Misiones, Córdoba y Buenos Aires de por lo menos tres individuos por punto. Estos individuos serán evaluados mediante técnicas de SSR para determinar mediante distintos parámetros la variabilidad intra e interpoblacional

The Pervasive Effects of ER Stress on a Typical Endocrine Cell: Dedifferentiation, Mesenchymal Shift and Antioxidant Response in the Thyrocyte

none13noThe endoplasmic reticulum stress and the unfolded protein response are triggered following an imbalance between protein load and protein folding. Until recently, two possible outcomes of the unfolded protein response have been considered: life or death. We sought to substantiate a third alternative, dedifferentiation, mesenchymal shift, and activation of the antioxidant response by using typical endocrine cells, i.e. thyroid cells. The thyroid is a unique system both of endoplasmic reticulum stress (a single protein, thyroglobulin represents the majority of proteins synthesized in the endoplasmic reticulum by the thyrocyte) and of polarized epithelium (the single layer of thyrocytes delimiting the follicle). Following endoplasmic reticulum stress, in thyroid cells the folding of thyroglobulin was disrupted. The mRNAs of unfolded protein response were induced or spliced (X-box binding protein-1). Differentiation was inhibited: mRNA levels of thyroid specific genes, and of thyroid transcription factors were dramatically downregulated, at least in part, transcriptionally. The dedifferentiating response was accompanied by an upregulation of mRNAs of antioxidant genes. Moreover, cadherin-1, and the thyroid (and kidney)-specific cadherin-16 mRNAs were downregulated, vimentin, and SNAI1 mRNAs were upregulated. In addition, loss of cortical actin and stress fibers formation were observed. Together, these data indicate that ER stress in thyroid cells induces dedifferentiation, loss of epithelial organization, shift towards a mesenchymal phenotype, and activation of the antioxidant response, highlighting, at the same time, a new and wide strategy to achieve survival following ER stress, and, as a sort of the other side of the coin, a possible new molecular mechanism of decline/loss of function leading to a deficit of thyroid hormones formation.openUlianich L.; Mirra P.; Garbi C.; Cali G.; Conza D.; Treglia A.S.; Miraglia A.; Punzi D.; Miele C.; Raciti G.A.; Beguinot F.; Consiglio E.; Di Jeso B.Ulianich, L.; Mirra, P.; Garbi, C.; Cali, G.; Conza, D.; Treglia, A. S.; Miraglia, A.; Punzi, D.; Miele, C.; Raciti, G. A.; Beguinot, F.; Consiglio, E.; Di Jeso, B

Estudios de diversidad genética en Salix humboldtiana

Salix humboldtiana Willd, vulgarmente conocido como «Sauce Criollo» es la única especie de sauce nativa de América del Sur. Su distribución en Argentina abarca desde Salta, Jujuy y Formosa hasta la Patagonia (Ragonese, 1966)1. Esta especie, presenta diversos usos: maderero, medicinal, ornamental, forrajera para ganado, melífera y para restauración de zonas ribereñas erosionadas. En las riberas e islas del Río Paraná, Paraguay y tributarios se reproduce naturalmente. Sin embargo, debido a la presencia de especies exóticas de sauces introducidas en su área de distribución natural, y a la fácil hibridación con las mismas amenazan la persistencia de la información genética del sauce nativo. Este proyecto tiene como objetivo estudiar la diversidad genética de la especie a través de marcadores microsatélites con el fin de rescatar individuos puros útiles para los programas de conservación y mejora. El programa de mejoramiento de esta especie se encuentra en una etapa inicial que consiste en introducciones de estacas, propagación y análisis genético de los individuos. Se han estudiado hasta el momento un total de 30 genotipos de la región del Delta del Paraná enVictoria, Entre Ríos, y en Formosa. A fin de conocer la variabilidad existente en estos materiales y la presencia de híbridos se analizaron los ADNs provenientes de hojas mediante marcadores moleculares microsatélites. La metodología empleada fue la reacción en cadena de la enzima polimerasa (PCR) y los fragmentos obtenidos fueron visualizados mediante electroforesis en geles de poliacrilamida. Se obtuvieron productos de amplificación para los 20 marcadores analizados y dentro del rango esperado resultando 14 marcadores monomórficos y 6 polimórficos. Además, la aplicación de marcadores diagnósticos sugirió que las muestras analizadas corresponderían a individuos de Salix humboldtiana puros y diploides. Actualmente se continúa trabajando con la puesta a punto de nuevos marcadores y con la extracción de ADN de individuos provenientes de Diamante, Entre Ríos.

Accuracy of dementia diagnosis—a direct comparison between radiologists and a computerized method

There has been recent interest in the application of machine learning techniques to neuroimaging-based diagnosis. These methods promise fully automated, standard PC-based clinical decisions, unbiased by variable radiological expertise. We recently used support vector machines (SVMs) to separate sporadic Alzheimer's disease from normal ageing and from fronto-temporal lobar degeneration (FTLD). In this study, we compare the results to those obtained by radiologists. A binary diagnostic classification was made by six radiologists with different levels of experience on the same scans and information that had been previously analysed with SVM. SVMs correctly classified 95% (sensitivity/specificity: 95/95) of sporadic Alzheimer's disease and controls into their respective groups. Radiologists correctly classified 65–95% (median 89%; sensitivity/specificity: 88/90) of scans. SVM correctly classified another set of sporadic Alzheimer's disease in 93% (sensitivity/specificity: 100/86) of cases, whereas radiologists ranged between 80% and 90% (median 83%; sensitivity/specificity: 80/85). SVMs were better at separating patients with sporadic Alzheimer's disease from those with FTLD (SVM 89%; sensitivity/specificity: 83/95; compared to radiological range from 63% to 83%; median 71%; sensitivity/specificity: 64/76). Radiologists were always accurate when they reported a high degree of diagnostic confidence. The results show that well-trained neuroradiologists classify typical Alzheimer's disease-associated scans comparable to SVMs. However, SVMs require no expert knowledge and trained SVMs can readily be exchanged between centres for use in diagnostic classification. These results are encouraging and indicate a role for computerized diagnostic methods in clinical practice

GRP78 Mediates Cell Growth and Invasiveness in Endometrial Cancer.

Abstract Recent studies have indicated that endoplasmic reticulum stress, the unfolded protein response activation and altered GRP78 expression can play an important role in a variety of tumors development and progression. Very recently we reported for the first time that GRP78 is increased in endometrial tumors. However, whether GRP78 could play a role in the growth and/or invasiveness of endometrial cancer cells is still unknown. Here we report that the silencing of GRP78 expression affects both cell growth and invasiveness of Ishikawa and AN3CA cells, analyzed by the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and transwell migration assay, respectively. At variance with Ishikawa cells, AN3CA cells showed, besides an endoplasmic reticulum, also a plasma membrane GRP78 localization, evidenced by both immunofluorescence and cell membrane biotinylation experiments. Intriguingly, flow cytometry experiments showed that the treatment with a specific antibody targeting GRP78 C-terminal domain caused apoptosis in AN3CA but not in Ishikawa cells. Induction of apoptosis in AN3CA cells was not mediated by the p53 pathway activation but was rather associated to reduced AKT phosphorylation. Interestingly, immunofluorescence analysis evidenced that endometrioid adenocarcinoma tissues displayed, similarly to AN3CA cells, also a GRP78 plasma membrane localization. These data suggest that GRP78 and its plasma membrane localization, might play a role in endometrial cancer development and progression and might constitute a novel target for the treatment of endometrial cancer

Sexual and individual foraging segregation in Gentoo penguins Pygoscelis papua from the Southern Ocean during an abnormal winter

Knowledge about sexual segregation and gender-specific, or indeed individual specialization, in marine organisms has improved considerably in the past decade. In this context, we tested the “Intersexual Competition Hypothesis” for penguins by investigating the feeding ecology of Gentoo penguins during their austral winter non-breeding season. We considered this during unusual environmental conditions (i.e. the year 2009 had observations of high sea surface and air temperatures) in comparison with the long term average at Bird Island, South Georgia. Through conventional (i.e. stomach contents) and stable isotopic values from red blood cells, plasma and feathers of both male and female Gentoo penguins, we showed that there were significant differences between sexes, with males feeding mainly on fish (54% by mass) followed by crustaceans (38%) whereas females fed mainly on crustaceans (89% by mass) followed by fish (4%). Themisto gaudichaudii was the most important crustacean prey for males (64% by mass; 82% by number; 53% by frequency of occurrence) and females (63% by mass; 77% by number; 89% by frequency of occurrence), contrasting with all previous studies that found Antarctic krill Euphausia superba were generally the main prey. Stable isotopic data showed that, in terms of habitat use (based on δ 13C), there were significant differences in short-term carbon signatures between males and females (based on plasma and red blood cells), suggesting that both sexes explored different habitats, with females exploring more offshore pelagic waters and males feeding more in coastal benthic waters. Based on δ 15N, males fed on significantly higher trophic level than females (based on plasma and red blood cells), in agreement with our diet results., Thus, Gentoo penguins behave in a similar manner to other non-breeding penguins species (e.g. king, macaroni and rockhopper penguins), albeit at a smaller spatial scale (as they do not disperse as these other penguins do), in that they have a wider habitat and trophic niche during the Antarctic Winter (in comparison to Summer). We also detected individual specialization in feeding/trophic levels for each gender, with certain males feeding mainly on fish and certain females mainly on crustaceans, which may be driven the prevailing environmental conditions that lead individuals to search for alternative prey, and cause sexual diet segregation. Our results provide further information to help improve understanding about sexual segregation and individual specialization of marine organisms, while contributing valuable information on the winter diet for Antarctic monitoring programs and for modelling Antarctic marine food webs