Massively Parallel Haplotyping on Microscopic Beads for the High-Throughput Phase Analysis of Single Molecules

In spite of the many advances in haplotyping methods, it is still very difficult to characterize rare haplotypes in tissues and different environmental samples or to accurately assess the haplotype diversity in large mixtures. This would require a haplotyping method capable of analyzing the phase of single molecules with an unprecedented throughput. Here we describe such a haplotyping method capable of analyzing in parallel hundreds of thousands single molecules in one experiment. In this method, multiple PCR reactions amplify different polymorphic regions of a single DNA molecule on a magnetic bead compartmentalized in an emulsion drop. The allelic states of the amplified polymorphisms are identified with fluorescently labeled probes that are then decoded from images taken of the arrayed beads by a microscope. This method can evaluate the phase of up to 3 polymorphisms separated by up to 5 kilobases in hundreds of thousands single molecules. We tested the sensitivity of the method by measuring the number of mutant haplotypes synthesized by four different commercially available enzymes: Phusion, Platinum Taq, Titanium Taq, and Phire. The digital nature of the method makes it highly sensitive to detecting haplotype ratios of less than 1∶10,000. We also accurately quantified chimera formation during the exponential phase of PCR by different DNA polymerases

La Grande Peur dans la montagne ou o nascimento de uma lenda

Ramuz era apaixonado pelas lendas alpinas e nelas se inspirou para escrever numerosos textos. O presente artigo examina o caso de La Grande Peur dans la montagne, de 1926, e mostra que o romance é atravessado por vários lugares comuns lendários, como o caçador diabólico e o purgatório glacial onde erram as almas danadas. À maneira de certas novelas de Maupassant, o personagem principal de La Grande Peur, Joseph, confronta-se com acontecimentos sobrenaturais que o fazem progressivamente penetrar em um universo onírico, fantástico, até mesmo fantasmático. O final do romance, ao menos na edição original, permite pensar que a narrativa como um todo é uma lenda

Stand-up-comedy inspired experiential learning for connecting emotions and cognitions in healthcare education ::a pilot study

In stand-up comedy, a single actor establishes an intimate relationship with the audience and discusses out loud emotional issues regarding taboo subjects. The ‘stand-up design of instruction’ (STUDI) uses similar techniques in a three minutes video to help learners connect emotional and cognitive experiences of mental health problems in six steps. The present article discusses a pilot experiment using the STUDI pedagogical model. The results showed that learners developed a positive appreciation of the training, a better emotional and cognitive understanding of the experience of mental health problems, the ability to exercise subjective and objective analysis through engaging in dialogue with other students, and increased motivation and memory-anchoring. In addition to other methods, such as simulated patients or peer teaching, STUDI can provide a simple and inexpensive pedagogical alternative using standing techniques to address large groups of students and short-circuit the barriers between emotions and cognitions

«Stand-up Design of Instruction» (STUDI) ::le stand-up comme modèle pédagogique en santé mentale

La psychiatrie ne doit ni se détacher de son ancrage médical et scientifique ni s’appauvrir de sa dimension psychothérapeutique: une discipline médicale où le corps et le vécu de la personne sont également considérés de manière indissociable. La proposition de stand-up permet de faire évoluer les pratiques pédagogiques de présentation de patients dans une perspective plus actuelle et plus éthique. Les modèles d’enseignement et d’évaluation de l’enseignement doivent ainsi être adaptés pour se rapprocher de cet objectif plutôt que de s’en éloigner

Multigene evidence for close evolutionary relations between Gromia and Foraminifera

Gromia oviformis is a common marine rhizopod, possessing a large ovoid membraneous theca that resembles the tests of certain monothalamous (single-chambered) foraminifers. In fact, the genus Gromia was initially classified among the Foraminifera, but because of its non-granular, filose pseudopodia it was later included among filopodia-bearing protists (the Filosea). Recent molecular phylogenies suggested that Gromia branches among Cercozoa, a heterogeneous assemblage of mainly amoeboid protists, which show some affinities to Foraminifera. To test how closely related are Gromia and Foraminifera, we have analysed the sequences of actin, large subunit of the RNA polymerase II (RPB1) and small subunit (SSU) rRNA genes. We also analysed the structure of the polyubiquitin gene of G. oviformis. Our analyses show that Gromia’s actin is specifically related to one of the two actin genes families known in Foraminifera. In RPB1-based phylogenies, Gromia appears as the closest relative of Foraminifera, while in the SSU rRNA trees it branches as sister to Foraminifera and Haplosporidia. We identified also a single serine insertion in the polyubiquitin of Gromia, similar to that found in Foraminifera, Plasmodiophorida and some Cercozoa. Altogether, these findings support the hypothesis that the morphological resemblance between Gromia and Foraminifera may be due to a shared common ancestor. If further analyses of protein-coding genes including a more representative sampling of Cercozoa confirm this relationship, then the molecular study of G. oviformis will be of key importance for understanding the origin of Foraminifera

Kinetic GFR Outperforms CKD-EPI for slow graft function prediction in the immediate postoperative period following kidney transplantation

Rapid identification of patients at high risk for slow graft function (SGF) is of major importance in the immediate period following renal graft transplantation, both for early therapeutic decisions and long-term prognosis. Due to the high variability of serum creatinine levels after surgery, glomerular filtration rate (GFR) estimation is challenging. In this situation, kinetic estimated GFR (KeGFR) equations are interesting tools but have never been assessed for the identification of SGF patients

Prognostic Scores for Ursodeoxycholic Acid-Treated Patients Predict Graft Loss and Mortality in Recurrent Primary Biliary Cholangitis after Liver Transplantation

Background/aim Recurrent primary biliary cholangitis (rPBC) develops in approximately 30% of patients and negatively impacts graft and overall patient survival after liver transplantation (LT). There is a lack of data regarding the response rate to ursodeoxycholic acid (UDCA) in rPBC. We evaluated a large, international, multi-center cohort to assess the performance of scores for PBC to predict the risk of graft and overall survival after LT in patients with rPBC. Methods A total of 332 patients with rPBC after LT were evaluated from 28 centres across Europe, North and South America. The median age at the time of rPBC was 58.0 years [IQR 53.2 - 62.6], and 298 patients (90%) were females. The biochemical response was measured with serum levels of alkaline phosphatase (ALP) and bilirubin, and Paris-2, GLOBE and UK-PBC scores at 1 year after UDCA initiation. Results During a median follow-up of 8.7 years [IQR 4.3 - 12.9] after rPBC diagnosis, 52 patients (16%) had graft loss and 103 (31%) died. After 1 year of UDCA initiation the histological stage at rPBC (HR, 3.97, 95%CI 1.36-11.55, P=0.01), use of prednisone (HR 3.18, 95%CI 1.04-9.73, P=0.04), ALP xULN (HR 1.59, 95%CI 1.26-2.01,

Prognostic Scores for Ursodeoxycholic Acid-Treated Patients Predict Graft Loss and Mortality in Recurrent Primary Biliary Cholangitis after Liver Transplantation

Background/aim Recurrent primary biliary cholangitis (rPBC) develops in approximately 30% of patients and negatively impacts graft and overall patient survival after liver transplantation (LT). There is a lack of data regarding the response rate to ursodeoxycholic acid (UDCA) in rPBC. We evaluated a large, international, multi-center cohort to assess the performance of scores for PBC to predict the risk of graft and overall survival after LT in patients with rPBC. Methods A total of 332 patients with rPBC after LT were evaluated from 28 centres across Europe, North and South America. The median age at the time of rPBC was 58.0 years [IQR 53.2 - 62.6], and 298 patients (90%) were females. The biochemical response was measured with serum levels of alkaline phosphatase (ALP) and bilirubin, and Paris-2, GLOBE and UK-PBC scores at 1 year after UDCA initiation. Results During a median follow-up of 8.7 years [IQR 4.3 - 12.9] after rPBC diagnosis, 52 patients (16%) had graft loss and 103 (31%) died. After 1 year of UDCA initiation the histological stage at rPBC (HR, 3.97, 95%CI 1.36-11.55, P=0.01), use of prednisone (HR 3.18, 95%CI 1.04-9.73, P=0.04), ALP xULN (HR 1.59, 95%CI 1.26-2.01,