Advanced Molecular Characterization Using Digital Spatial Profiling Technology on Immunooncology Targets in Methylated Compared with Unmethylated IDH-Wildtype Glioblastoma.

INTRODUCTION: Glioblastoma (GBM) is the most common primary adult brain tumour with a median overall survival (OS) of 12-15 months. Molecular characterization of multiple immunooncology targets in GBM may help target novel immunotherapeutic strategies. We used NanoString GeoMx® Digital Spatial Profiling (DSP) to assess multiple immunooncology protein targets in methylated versus unmethylated IDH-wild-type glioblastoma. METHODS: NanoString GeoMx® DSP technology uses multiple primary antibodies conjugated to indexing DNA oligos with a UV photocleavable linker. Tissue regions of interest (ROIs) are selected with bound fluorescent antibodies; oligos are released via a UV-mediated linker and quantitated. We used DSP multiplex analysis of 31 immunooncology proteins and controls (CD4, CD14, CD68, CD8A, B7-H3, PD-L1, CD19, FOXP3, CD44, STAT3 (phospho Y705), CD45, Pan Cytokeratin, MS4A1/CD20, CD45RO, PD1, CD3, beta-2 microglobulin, VISTA, Bcl2, GZMB, PTEN, beta-catenin, CD56, Ki-67, STAT3, AKT, p-Akt, S6, Histone H3, IgG Rabbit control, and Mouse IgG control) from ROIs in a cohort of 10 IDH-wild-type glioblastomas (5 methylated and 5 unmethylated). An nCounter platform allowed quantitative comparisons of antibodies between ROIs in MGMT methylated and unmethylated tumours. Mean protein expression counts between methylated and unmethylated GBM were compared using technical and biological replicates. RESULTS: The analysis showed 10/27 immunooncology target proteins were significantly increased in methylated versus unmethylated IDH-wild-type glioblastoma tumour core (false discovery rate (FDR) <0.1 by Benjamini-Hochberg procedure). CONCLUSIONS: NanoString GeoMx® DSP was used to analyse multiple immunooncology protein target expression in methylated versus unmethylated IDH-wild-type glioblastoma. In this small study, there was a statistical increase in CD4, CD14, CD68, CD8A, B7-H3, PDL-1, CD19, FOXP3, CD44, and STAT3 protein expression in methylated versus unmethylated GBM tumour core; however, this requires larger cohort validation. Advanced multiplex immunooncological biomarker analysis may be useful in identifying biomarkers for novel immunotherapeutic agents in GBMs.

[Venezuelan consensus guidelines on the use of magnetic resonance in diagnosis and follow up of patients with multiple sclerosis]. / Consenso venezolano para el uso de la resonancia magnetica en el diagnostico y seguimiento de pacientes con esclerosis multiple.

INTRODUCTION: The clinical use of magnetic resonance (MR) in patients with multiple sclerosis (MS) has advanced markedly over the past few years. Several groups around the world have developed consensus guidelines about the role of MR in MS at diagnosis and during follow up. However, in some regions is difficult to extrapolate the recommendations. AIM: To provide recommendations for the implementation of MR in MS patients at diagnosis and follow up in Venezuela. DEVELOPMENT: A group of experts from Venezuela that included neurologists and radiologists, by using the online surveys methodology as well as face to face meetings developed the intended consensus for the use of MR during the diagnosis and follow up of MS patients in Venezuela. Seventeen recommendations were established based on published evidence and the expert opinion. Recommendations focused on the role of conventional MR techniques and brain atrophy measurement in MS patients both at diagnosis and during follow-up. CONCLUSIONS: The recommendations of this consensus guidelines attempts to optimize the health care and management of patients with MS in Venezuela.

TITLE: Consenso venezolano para el uso de la resonancia magnetica en el diagnostico y seguimiento de pacientes con esclerosis multiple.Introduccion. El uso de la resonancia magnetica (RM) en el diagnostico y seguimiento de pacientes con esclerosis multiple (EM) se ha incrementado considerablemente durante los ultimos años. Diversos grupos de trabajo internacionales han intentado clarificar y normativizar el uso de la RM tanto en el momento del diagnostico como durante el seguimiento de los pacientes. Sin embargo, en muchas ocasiones se extrapolan datos de otras regiones que no contemplan la realidad de cada lugar o son dificiles de implementar. Objetivo. Elaborar un consenso venezolano para el uso de la RM en el diagnostico y seguimiento de pacientes con EM. Desarrollo. Un grupo de expertos de Venezuela, conformado por neurologos y radiologos, mediante metodologia de ronda de encuestas a distancia y reuniones presenciales, llevo adelante la elaboracion del consenso pretendido para el uso de la RM en el diagnostico y seguimiento de pacientes con EM en Venezuela. Se establecieron 17 recomendaciones basadas en la evidencia publicada y en el criterio de los expertos que participaron. Las recomendaciones se enfocaron en el papel de las tecnicas convencionales de RM, asi como en el de la medicion de la atrofia cerebral en pacientes con EM, tanto en el momento del diagnostico como durante el seguimiento. Conclusion. Las recomendaciones establecidas en el presente consenso permitiran optimizar el cuidado y el seguimiento de los pacientes con EM en Venezuela.

Stochastic thermodynamics of fluctuating density fields: non-equilibrium free energy differences under coarse-graining.

We discuss the stochastic thermodynamics of systems that are described by a time-dependent density field, for example, simple liquids and colloidal suspensions. For a time-dependent change of external parameters, we show that the Jarzynski relation connecting work with the change of free energy holds if the time evolution of the density follows the Kawasaki-Dean equation. Specifically, we study the work distributions for the compression and expansion of a two-dimensional colloidal model suspension implementing a practical coarse-graining scheme of the microscopic particle positions. We demonstrate that even if coarse-grained dynamics and density functional do not match, the fluctuation relations for the work still hold albeit for a different, apparent, change of free energy.

Noninvasive measurement of dissipation in colloidal systems.

According to Harada and Sasa [Phys. Rev. Lett. 95, 130602 (2005)], heat production generated in a nonequilibrium steady state can be inferred from measuring response and correlation functions. In many colloidal systems, however, it is a nontrivial task to determine response functions, whereas details about spatial steady state trajectories are easily accessible. Using a simple conditional averaging procedure, we show how this fact can be exploited to reliably evaluate average heat production. We test this method using Brownian dynamics simulations, and apply it to experimental data of an interacting driven colloidal system.

Role of hidden slow degrees of freedom in the fluctuation theorem.

The validity of the fluctuation theorem for entropy production as deduced from the observation of trajectories implicitly requires that all slow degrees of freedom are accessible. We experimentally investigate the role of hidden slow degrees of freedom in a system of two magnetically coupled driven colloidal particles. The apparent entropy production based on the observation of just one particle obeys a fluctuation theorem-like symmetry with a slope of 1 in the short time limit. For longer times, we find a constant slope, but different from 1. We present theoretical arguments for a generic linear behavior both for small and large apparent entropy production but not necessarily throughout. By fine-tuning experimental parameters, such an intermediate nonlinear behavior can indeed be recovered in our system as well.

Chromosome aberrations in pesticide-exposed greenhouse workers.

OBJECTIVES: The aim of this study was to investigate the possibility of subtoxic exposure to pesticides causing chromosome aberrations in greenhouse workers. METHODS: In a cross-sectional and prospective study design chromosome aberration frequencies in cultured lymphocytes were examined for 116 greenhouse workers exposed to a complex mixture of almost 50 insecticides, fungicides, and growth regulators and also for 29 nonsmoking, nonpesticide-exposed referents. RESULTS: The preseason frequencies of chromosome aberrations were slightly but not statistically significantly elevated for the greenhouse workers when they were compared with the referents. After a summer season of pesticide spraying in the greenhouses, the total frequencies of cells with chromosome aberrations were significantly higher than in the preseason samples (P=0.02) and also higher than for the referents (P=0.05). This finding was especially due to an increased number of cells with chromatid gaps between the first and second samples (P=0.001). The results may reflect an additive genotoxic effect of the spraying season, for which the use of insecticides and growth regulators (but not fungicides) culminates. The highest elevation in the risk of chromatid gaps was observed for persons who did not use gloves during re-entry activities such as nipping, cutting, pricking, and potting (risk ratio 2.88, 95% confidence interval 1.63-5.11). CONCLUSIONS: The present results suggest a genotoxic effect from a complex subtoxic occupational pesticide exposure. In general, the findings indicate the importance of personal protection, during high-exposure re-entry activities, in preventing pesticide uptake and genetic damage.

[Complications of laparoscopic cholecystectomy]. / Complicaciones de la colecistectomía laparoscópica.

The laparoscopy cholecistectomy is a surgical procedure described in 1987, and it has had an important apogee and it's had substitute to open procedure. Between its advantages is notable, the postoperatory evolution without pain, paralytic ileus and the short time of hospitalization besides the patient can go away the same day in some cases. In Venezuela, the procedure has been received with enthusiasm by the surgeons with prefer this procedure however, there are complications derived in part for the lack of experience in the first cases, but occur non related of the experience. The quantization of the complications is not simple to realize for different reasons. We present twelve complications, two belong to patients intervened in the Hospital Universitario de Caracas and ten belong to patient transferred from other center. The complications were: pseudoaneurism of hepatic artery, hematoma of the wall, cutting of common bile duct, section the right bile duct, abscess of vesicular bed in two cases, retropneumoperitoneum, ascites by biliary fistula and four biloma. The treatment varied in every case and the utilization of radiologic technique with percutaneous drainage were useful in five cases. We conclude: first, this technique is not free of complications. Second, the percutaneous drainage (abscess, biloma), endoscopic procedure (biliary prosthesis) in fistulaes and arteriographics (arterial embolization), are useful in some cases, and could prevent the surgical reintervention.

[Potassium-dependent modification of the lidocaine effect: an experimental investigation (author's transl)]. / Tierexperimentelle Untersuchungen zur Frage einer kaliumabhängigen Modifikation der Lidocainwirkung

Lidocaine was supplied to 10 cats by 2 comparative infusions of 7.5 mg/kg respectively. Once the drug was dissolved in a physiological and the other time in a K-Mg-concentrated electrolyte solution. The sequence of both infusions alternated. Serum potassium remained constant during infusion of the physiological electrolyte solution, but increased by about 1 mEqu/1 under K-concentrated solution. Lidocaine caused an enhancement of ventricular fibrillation threshold to an absolutely identical degree in both groups. Increase in cardiac output and stroke volume and decrease of sinus rate was significant in both groups, but more pronounced under K-Mg concentrated solution. Mean arterial pressure, peripheral vascular resistance, PQ, QRS, and QT interval remained constant on the whole. We deduce from these results for antiarrhythmic therapy on normokalemic patients, that the addition of potassium and magnesium to lidocaine solution does not enhance its antifibrillatory effect. This statement is valid for doses up to 20 mEqu/h of magnesium and 40 mEqu/h of potassium.