A Database of Accurate Electrophoretic Migration Patterns for Human Proteins.

Native molecular weight (MW) is one of the defining features of proteins. Denaturing gel electrophoresis (SDS-PAGE) is a very popular technique for separating proteins and determining their MW. Coupled with antibody-based detection, SDS-PAGE is widely applied for protein identification and quantitation. Yet, electrophoresis is poorly reproducible and the MWs obtained are often inaccurate. This hampers antibody validation and negatively impacts the reliability of western blot data, resulting worldwide in a considerable waste of reagents and labour. We argue that, to alleviate these problems there is a need to establish a database of reference MWs measured by SDS-PAGE. Using mass spectrometry as an orthogonal detection method, we acquired electrophoretic migration patterns for approximately 10'000 human proteins in five commonly used cell lines. We applied a robust internal calibration of migration to determine accurate and reproducible molecular weights. This in turn allows merging replicates to increase accuracy, but also enables comparing different cell lines. Mining of the data obtained highlights structural factors that affect migration of distinct classes of proteins. When combined with peptide coverage, the data produced recapitulates known post-translational modifications and differential splicing and can be used to formulate hypotheses on new or poorly known processing events. The full information is freely accessible as a web resource through a user friendly graphical interface (https://pumba.dcsr.unil.ch/). We anticipate that this database will be useful to investigators worldwide for troubleshooting western blot experiments, but could also contribute to the characterization of human proteoforms.

APOBEC3C, a nucleolar protein induced by genotoxins, is excluded from DNA damage sites.

The human genome contains 11 APOBEC (apolipoprotein B mRNA editing catalytic polypeptide-like) cytidine deaminases classified into four families. These proteins function mainly in innate antiviral immunity and can also restrict endogenous retrotransposable element multiplication. The present study focuses on APOBEC3C (A3C), a member of the APOBEC3 subfamily. Some APOBEC3 proteins use their enzymatic activity on genomic DNA, inducing mutations and DNA damage, while other members facilitate DNA repair. Our results show that A3C is highly expressed in cells treated with DNA-damaging agents. Its expression is regulated by p53. Depletion of A3C slightly decreases proliferation and does not affect DNA repair via homologous recombination or nonhomologous end joining. The A3C interactomes obtained from control cells and cells exposed to the genotoxin etoposide indicated that A3C is a nucleolar protein. This was confirmed by the detection of either endogenous or ectopic A3C in nucleoli. Interestingly, we show that A3C is excluded from areas of DNA breaks in live cells. Our data also indicate that the C-terminal part of A3C is responsible for its nucleolar localization and exclusion from DNA damage sites.

La inflamación como el nexo entre la enfermedad periodontal y la enfermedad cardiovascular / Inflammation as a link between periodontal disease and cardiovascular disease

La periodontitis es una enfermedad no transmisible, con una alta prevalencia, que oscila entre el 45% y el 50% de la población mundial, ocupando el sexto lugar entre las enfermedades más frecuentes de la huma-nidad. Existe suficiente evidencia que avala la relación entre la enfermedad periodontal y la enfermedad car-diovascular, responsable de aproximadamente el 45% de las muertes en países desarrollados, compren-diendo en su causalidad al infarto de miocardio, el accidente cerebrovascular, la insuficiencia cardíaca y las arritmias, que causan alrededor del 95 % de las muertes relacionadas con la enfermedad cardiovas-cular. Ambas patologías presentan factores de riesgo comunes ampliamente reconocidos, como la diabetes y el tabaquismo, pero además manifiestan caracte-rísticas genéticas y epigenéticas que avalan distintos mecanismos etiopatológicos. Más allá de los factores de riesgo comunes, se han propuesto dos mecanis-mos para explicar la relación entre la enfermedad periodontal y las cardiovasculares. Uno de ellos, constituye la invasión directa de patógenos periodontales en las células endoteliales. El otro mecanismo sugerido (vía indirecta), ocasionado por la respuesta inflamatoria sistémica que resulta en niveles cróni-camente elevados de diferentes citoquinas, también relacionadas con la enfermedad vascular aterosclerotica como IL-1ß, IL-6, IL-8, TNF-α, PCR y la proteína quimioatrayente de monocitos, podría estar mediado por productos bacterianos, como los lipopolisacári-dos que alcanzarían la circulación induciendo una potente respuesta inmunitaria. Estos mecanismos pueden actuar inflamando las células endoteliales, modulando el metabolismo de los lípidos y aumentan-do el estrés oxidativo, favoreciendo la aterosclerosis, conformando la expresión de un fenotipo arterial in-flamatorio, generando el nexo entre la enfermedad periodontal y las patologías cardiovasculares (AU))

Periodontitis is a non-communicable disease which is highly prevalent worldwide. It was reported to range from 45% to 50% around the world and it was the sixth most prevalent condition of humanity. Consistent body of evidence explains the relationship between periodontal disease and other common systemic conditions such as cardiovascular disease. Periodontitis is likely to cause a 45% of deaths in developed countries, including myocardial infarction, stroke, heart failure and arrhythmias that cause about a 95% of deaths related to cardiovascular disease.Both diseases share many risk factors, such as diabetes and smoking; but also, genetic, and epigenetic characteristics support several etiopathological mechanisms. Beyond the common risk factors, two mechanisms have been proposed to elucidate the relationship between the periodontal disease and cardiovascular diseases. One of them supports the concept that periodontal pathogens are capable of the direct invasion of endothelial cells. The other mechanism suggested (indirect pathway), caused by the disease resulting in chronically elevation of CRP, inflammatory cytokines, the monocyte chemoattractant protein, could be mediated by bacterial products, such as lipopolysaccharides, wich induce a potent immune response and can accelerate endothelial dysfunction. These mechanisms may act by inflaming endothelial cells, modulating lipid metabolism and increasing oxidative stress, favoring atherosclerosis, determining the expression of an inflammatory arterial phenotype, generating the link between periodontal disease and cardiovascular pathologies (AU)

The proteolytic landscape of cells exposed to non-lethal stresses is shaped by executioner caspases.

Cells are in constant adaptation to environmental changes to insure their proper functioning. When exposed to stresses, cells activate specific pathways to elicit adaptive modifications. Those changes can be mediated by selective modulation of gene and protein expression as well as by post-translational modifications, such as phosphorylation and proteolytic processing. Protein cleavage, as a controlled and limited post-translational modification, is involved in diverse physiological processes such as the maintenance of protein homeostasis, activation of repair pathways, apoptosis and the regulation of proliferation. Here we assessed by quantitative proteomics the proteolytic landscape in two cell lines subjected to low cisplatin concentrations used as a mild non-lethal stress paradigm. This landscape was compared to the one obtained in the same cells stimulated with cisplatin concentrations inducing apoptosis. These analyses were performed in wild-type cells and in cells lacking the two main executioner caspases: caspase-3 and caspase-7. Ninety-two proteins were found to be cleaved at one or a few sites (discrete cleavage) in low stress conditions compared to four hundred and fifty-three in apoptotic cells. Many of the cleaved proteins in stressed cells were also found to be cleaved in apoptotic conditions. As expected, ~90% of the cleavage events were dependent on caspase-3/caspase-7 in apoptotic cells. Strikingly, upon exposure to non-lethal stresses, no discrete cleavage was detected in cells lacking caspase-3 and caspase-7. This indicates that the proteolytic landscape in stressed viable cells fully depends on the activity of executioner caspases. These results suggest that the so-called executioner caspases fulfill important stress adaptive responses distinct from their role in apoptosis. Mass spectrometry data are available via ProteomeXchange with identifier PXD023488.

Prognostic Factors of Children Admitted to a Pediatric Intensive Care Unit After an Episode of Drowning.

OBJECTIVE: The aim of this study was to evaluate the prognostic factors of patients admitted to a pediatric intensive care unit (PICU) after drowning. METHODS: Retrospective observational study from January 1992 to December 2004 and prospective study from January 2005 to December 2015 were conducted in a tertiary children's hospital PICU. The data analyzed refer to the patient, event, type of resuscitation performed, and clinical situation after resuscitation and at arrival to the PICU; results of additional tests; and clinical evolution and neurological status at discharge from the PICU (categorized as death, severe encephalopathy, or normal). The considered potential prognostic factors were whether drowning was witnessed, the type of initial resuscitation, Glasgow Coma Scale score at admission, pupil status and reactivity, and pH. RESULTS: One hundred thirty-one patients were registered. Mortality was 16.7%, and 8.3% had significant neurological sequelae. The clearest factor associated with poor outcome was the type of initial resuscitation performed. All patients who did not require cardiopulmonary resuscitation (CPR), or only basic CPR, had good outcomes; 96.3% of those who required advanced CPR with epinephrine administration had poor outcomes. Patients who needed advanced resuscitation with administration of epinephrine had lower temperature, Glasgow Coma Scale score, pH, and bicarbonate at admission and higher level of glucose. In this group, there was also a higher incidence of seizures, acute respiratory distress syndrome, hemodynamic compromise, and acute renal failure. CONCLUSIONS: The need for advanced CPR with epinephrine administration on the scene predicts poor neurological outcome (severe encephalopathy or death) in drowned children.

Consenso de hemofilia en México / Mexican Consensus in Haemophilia

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Abstract Hemophilia is a hemorrhagic disorder with a sex-linked inherited pattern, characterized by an inability to amplify coagulation due to a deficiency in coagulation factor VIII (hemophilia A or classic) or factor IX (hemophilia B). Sequencing of the genes involved in hemophilia has provided a description and record of the main mutations, as well as a correlation with the various degrees of severity. Hemorrhagic manifestations are related to levels of circulating factor, mainly affecting the musculoskeletal system and specifically the large joints (knees, ankles and elbows). This document is a review and consensus of the main genetic aspects of hemophilia, from the inheritance pattern to the concept of women carriers, physiopathology and classification of the disorder, the basic and confirmation studies when hemophilia is suspected, the various treatment regimens based on infusion of the deficient coagulation factor as well as innovative factor-free therapies and recommendations for the management of complications associated with treatment (development of inhibitors and/or transfusion transmitted infections) or secondary to articular hemorrhagic events (hemophilic arthropathy). Finally, relevant reviews of clinical and treatment aspects of hemorrhagic pathology charachterized by acquired deficiency of FVIII secondary to neutralized antibodies named acquired hemophilia.

Endoscopic ultrasound-guided fiducial placement in pancreatic tumors: safety and technical feasibility

Background and aims: stereotactic body radiation therapy (SBRT) for pancreatic malignancies requires the placement of fiducials to guide treatment delivery. The aim of this study was to assess the safety and feasibility of endoscopic ultrasound (EUS) guided fiducial placement using a 22-gauge needle, in patients with pancreatic cancer undergoing SBRT. Methods: this single-center retrospective study included 47 patients with biopsy-proven advanced pancreatic cancer who underwent EUS-guided fiducial placement between February 2014 and February 2018. Primary outcome measurements included technical success, fiducial migration rate and procedural complications. Results: all 47 patients received a sufficient number of fiducials and could therefore undergo a successful SBRT. The mean number of fiducials inserted per case was 2 +/- 1 (range 1-3) and no fiducial migration was noted. The adverse event rate was 4.2%, as one patient developed mild pancreatitis and another patient required one week of hospitalization one month after fiducial placement due to a duodenal abscess. Conclusions: EUS-guided fiducial placement is a safe and technically feasible procedure in centers with endosonographers that are well trained in EUS with FNA

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Endoscopic ultrasound-guided fiducial placement in pancreatic tumors: safety and technical feasibility.

BACKGROUND AND AIMS: stereotactic body radiation therapy (SBRT) for pancreatic malignancies requires the placement of fiducials to guide treatment delivery. The aim of this study was to assess the safety and feasibility of endoscopic ultrasound (EUS) guided fiducial placement using a 22-gauge needle, in patients with pancreatic cancer undergoing SBRT. METHODS: this single-center retrospective study included 47 patients with biopsy-proven advanced pancreatic cancer who underwent EUS-guided fiducial placement between February 2014 and February 2018. Primary outcome measurements included technical success, fiducial migration rate and procedural complications. RESULTS: all 47 patients received a sufficient number of fiducials and could therefore undergo a successful SBRT. The mean number of fiducials inserted per case was 2 ± 1 (range 1-3) and no fiducial migration was noted. The adverse event rate was 4.2%, as one patient developed mild pancreatitis and another patient required one week of hospitalization one month after fiducial placement due to a duodenal abscess. CONCLUSIONS: EUS-guided fiducial placement is a safe and technically feasible procedure in centers with endosonographers that are well trained in EUS with FNA.

Comparing the Performance of Indoor Localization Systems through the EvAAL Framework.

In recent years, indoor localization systems have been the object of significant research activity and of growing interest for their great expected social impact and their impressive business potential. Application areas include tracking and navigation, activity monitoring, personalized advertising, Active and Assisted Living (AAL), traceability, Internet of Things (IoT) networks, and Home-land Security. In spite of the numerous research advances and the great industrial interest, no canned solutions have yet been defined. The diversity and heterogeneity of applications, scenarios, sensor and user requirements, make it difficult to create uniform solutions. From that diverse reality, a main problem is derived that consists in the lack of a consensus both in terms of the metrics and the procedures used to measure the performance of the different indoor localization and navigation proposals. This paper introduces the general lines of the EvAAL benchmarking framework, which is aimed at a fair comparison of indoor positioning systems through a challenging competition under complex, realistic conditions. To evaluate the framework capabilities, we show how it was used in the 2016 Indoor Positioning and Indoor Navigation (IPIN) Competition. The 2016 IPIN competition considered three different scenario dimensions, with a variety of use cases: (1) pedestrian versus robotic navigation, (2) smartphones versus custom hardware usage and (3) real-time positioning versus off-line post-processing. A total of four competition tracks were evaluated under the same EvAAL benchmark framework in order to validate its potential to become a standard for evaluating indoor localization solutions. The experience gained during the competition and feedback from track organizers and competitors showed that the EvAAL framework is flexible enough to successfully fit the very different tracks and appears adequate to compare indoor positioning systems.

Effect of angiotensin-converting enzyme inhibitors on vascular endothelial function in hypertensive patients after intensive periodontal treatment / Efecto de los inhibidores de la enzima convertidora sobre la función del endotelio vascular en pacientes hipertensos que recibieron tratamiento periodontal intensivo

There is a relation between vascular endothelial function,atherosclerotic disease, and inflammation. Deterioration ofendothelial function has been observed twenty-four hours afterintensive periodontal treatment. This effect may be counteractedby the action of angiotensin-converting enzyme inhibitors, whichimprove endothelial function. The aim of the present study was toevaluate vascular endothelial function after intensive periodontaltreatment, in hypertensive patients treated with angiotensin-converting enzyme inhibitors. A prospective, longitudinal, comparative study involving repeatedmeasurements was conducted. Fifty-two consecutive patients withsevere periodontal disease were divided into two groups, onecomprising hypertensive patients treated with converting enzymeinhibitors and the other comprising patients with no clinical signsof pathology and not receiving angiotensin-converting enzymeinhibitors. Endothelial function was assessed by measuring post-ischemic dilation of the humeral artery (baseline echocardiographyDoppler), and intensive periodontal treatment was performed 24hlater. Endothelial function was re-assessed 24h and 15 days after periodontal treatment. Statistical analysis: Results were analyzedusing the SPSS 20 statistical software package. Student’s t test andMANOVA were calculated and linear regression analysis with 95%confidence intervals and α<0.05 was performed. Arterial dilationat 24 hours was lower compared to baseline in both groups; valuescorresponding to the groups receiving angiotensin-convertingenzyme inhibitors were 11.89 ± 4.87 vs. 7.30 ± 2.90% (p<0.01) andthose corresponding to the group not receiving ACE inhibitors were12.72 ± 4.62 vs. 3.56 ± 2.39 (p<0.001). The differences betweengroups were statistically significant (p<0.001)...

Existe relación entre la disfunción del endotelio vascular, la enfermedad ateroesclerótica y la inflamación. A las 24 horas del tratamiento intensivo de la enfermedad periodontal, se produce un deterioro de la función endotelial. Este efecto podría ser balanceado por la acción de los inhibidores de la enzima convertidora de la angiotensina que mejoran la función endotelial. El objetivo del presente estudio fue evaluar la función endotelial vascular después del tratamiento periodontal intensivo, en pacientes hipertensos tratados con inhibidores de la enzima convertidora de la angiotensina. Se realizó un estudio prospectivo, longitudinal, comparativo, con mediciones repetidas. Se incorporaron 52 pacientes con enfermedad periodontal severa divididos en dos grupos, uno con hipertensión arterial, tratados con inhibidores de la enzima convertidora y otro sin inhibidores ni patología clínicamente evidente. Se determinó la función endotelial cuantificando la dilatación de la arterial humeral post isquemia ecocardiografía Doppler basal. A las 24 hs se efectuó el tratamiento periodontal intensivo; a las 24 h y 15 días posteriores se reevaluó la función endotelial. Análisis estadístico: se empleó el paquete estadístico SPSS 20. Se realizaron: t-test de Student, MANOVA y análisis de regresión lineal con intervalos de confianza del 95 por ciento y a <0.05. Resultados: a las 24 h post tratamiento periodontal, se observó una menor dilatación arterial en ambos grupos, en relación a la dilatación arterial basal, siendo para el grupo con inhibidores 11.89 +- 4.87 vs. 7.30 +- 2.90 por ciento, p<0.01 y para el grupo sin inhibidores 12.72 +- 4.62 vs. 3.56 +- 2.39, p<0.001, con diferencias significativas entre ambos p<0.001...

Acoustic sensor network for relative positioning of nodes.

In this work, an acoustic sensor network for a relative localization system is analyzed by reporting the accuracy achieved in the position estimation. The proposed system has been designed for those applications where objects are not restricted to a particular environment and thus one cannot depend on any external infrastructure to compute their positions. The objects are capable of computing spatial relations among themselves using only acoustic emissions as a ranging mechanism. The object positions are computed by a multidimensional scaling (MDS) technique and, afterwards, a least-square algorithm, based on the Levenberg-Marquardt algorithm (LMA), is applied to refine results. Regarding the position estimation, all the parameters involved in the computation of the temporary relations with the proposed ranging mechanism have been considered. The obtained results show that a fine-grained localization can be achieved considering a Gaussian distribution error in the proposed ranging mechanism. Furthermore, since acoustic sensors require a line-of-sight to properly work, the system has been tested by modeling the lost of this line-of-sight as a non-Gaussian error. A suitable position estimation has been achieved even if it is considered a bias of up to 25 of the line-of-sight measurements among a set of nodes.

[Genotypes of Candida dubliniensis in clinical isolates]. / Genotipos de Candida dubliniensis en aislamientos clínicos.

Amplification of specific sequences of the ITS1 and ITS2 regions and the intervening 5.8S rRNA gene has lead to the identification of four separate genotypes in Candida dubliniensis. Using primers specific for each genotype, we have studied the prevalence of these genotypes among 68 clinical isolates, mostly from Spanish patients infected by HIV. The majority of the isolates tested belonged to genotype 1 (97%), while only one isolate each from genotypes 2 (1.5%) and 3 (1.5%) were detected in the oral cavity of two patients with HIV infection.

Evaluacion de la funcion sistolica y diastolica del ventriculo izquierdo en el infarto agudo de miocardio por ecograma aortico / Aortic echogram in the evaluation of systolic and diastolic left ventricular function in acute myocardial infarction

Se investigó en un grupo de pacientes portadores de infarto agudo de miocardio (IAM), un método incruento para la determinación del volumen minuto cardíaco (VM) y la función ventricular diastólica a partir del ecograma aórtico. Los parámetros utilizados para la determinación del VM fueron: el promedio de apertura inicial y final sistólica de la ssigmoideas (ASA), período eyectivo del ventrículo izquierdo (PE) amplitud del movimiento de la pared psoterior aórtica durante la eyección (AA). La fórmula empleada y que fuera previamente publicada es la siguiente: VM = (ASA x xPE) x 100 + AA x xFC. La función ventricular diastólica se evaluó utilizando el índice de vaciamiento auricular izquierdo (IVAI) descripto por Strunk y col. El mismo se expresa por la relación X?OA donde X corresponde al movimiento de la pared posterior aórtica (PPA) en el primer tercio de la diástole y OA comprende todo el movimiento de la PPA durante la diástole pasiva. A todos los pacientes se les realizaron determinaciones del VM por termodilución y registro de presión capilar (PC), obteniendose en total 20 mediciones en 14 pacientes. Para el VM se observó buena correlación lineal (r = 0,81). Respecto a la función ventricular diastólica, el IVAI alcanzó un valor de 0,49 (ñ 0,23) para un normal de 0,82 (p < 0,01) hallándose significativamente disminuido, evidenciando la rigidez de la cámara. Consideramos que la utilización del ecograma aórtico y el análisis de los movimientos de la PPA constituye un procdedimiento repetible y confiable que permite estimar la situación hemodinámica en el IAM

Evaluacion de la funcion sistolica y diastolica del ventriculo izquierdo en el infarto agudo de miocardio por ecograma aortico / Aortic echogram in the evaluation of systolic and diastolic left ventricular function in acute myocardial infarction

Se investigó en un grupo de pacientes portadores de infarto agudo de miocardio (IAM), un método incruento para la determinación del volumen minuto cardíaco (VM) y la función ventricular diastólica a partir del ecograma aórtico. Los parámetros utilizados para la determinación del VM fueron: el promedio de apertura inicial y final sistólica de la ssigmoideas (ASA), período eyectivo del ventrículo izquierdo (PE) amplitud del movimiento de la pared psoterior aórtica durante la eyección (AA). La fórmula empleada y que fuera previamente publicada es la siguiente: VM = (ASA x xPE) x 100 + AA x xFC. La función ventricular diastólica se evaluó utilizando el índice de vaciamiento auricular izquierdo (IVAI) descripto por Strunk y col. El mismo se expresa por la relación X?OA donde X corresponde al movimiento de la pared posterior aórtica (PPA) en el primer tercio de la diástole y OA comprende todo el movimiento de la PPA durante la diástole pasiva. A todos los pacientes se les realizaron determinaciones del VM por termodilución y registro de presión capilar (PC), obteniendose en total 20 mediciones en 14 pacientes. Para el VM se observó buena correlación lineal (r = 0,81). Respecto a la función ventricular diastólica, el IVAI alcanzó un valor de 0,49 (ñ 0,23) para un normal de 0,82 (p < 0,01) hallándose significativamente disminuido, evidenciando la rigidez de la cámara. Consideramos que la utilización del ecograma aórtico y el análisis de los movimientos de la PPA constituye un procdedimiento repetible y confiable que permite estimar la situación hemodinámica en el IAM (AU)