Advancing brain barriers RNA sequencing: guidelines from experimental design to publication

Background: RNA sequencing (RNA-Seq) in its varied forms has become an indispensable tool for analyzing differential gene expression and thus characterization of specific tissues. Aiming to understand the brain barriers genetic signature, RNA seq has also been introduced in brain barriers research. This has led to availability of both, bulk and single-cell RNA-Seq datasets over the last few years. If appropriately performed, the RNA-Seq studies provide powerful datasets that allow for significant deepening of knowledge on the molecular mechanisms that establish the brain barriers. However, RNA-Seq studies comprise complex workflows that require to consider many options and variables before, during and after the proper sequencing process.Main body: In the current manuscript, we build on the interdisciplinary experience of the European PhD Training Network BtRAIN (https://www.btrain-2020.eu/) where bioinformaticians and brain barriers researchers collaborated to analyze and establish RNA-Seq datasets on vertebrate brain barriers. The obstacles BtRAIN has identified in this process have been integrated into the present manuscript. It provides guidelines along the entire workflow of brain barriers RNA-Seq studies starting from the overall experimental design to interpretation of results. Focusing on the vertebrate endothelial blood–brain barrier (BBB) and epithelial blood-cerebrospinal-fluid barrier (BCSFB) of the choroid plexus, we provide a step-by-step description of the workflow, highlighting the decisions to be made at each step of the workflow and explaining the strengths and weaknesses of individual choices made. Finally, we propose recommendations for accurate data interpretation and on the information to be included into a publication to ensure appropriate accessibility of the data and reproducibility of the observations by the scientific community.Conclusion: Next generation transcriptomic profiling of the brain barriers provides a novel resource for understanding the development, function and pathology of these barrier cells, which is essential for understanding CNS homeostasis and disease. Continuous advancement and sophistication of RNA-Seq will require interdisciplinary approaches between brain barrier researchers and bioinformaticians as successfully performed in BtRAIN. The present guidelines are built on the BtRAIN interdisciplinary experience and aim to facilitate collaboration of brain barriers researchers with bioinformaticians to advance RNA-Seq study design in the brain barriers community

Synthesis and acidic properties of the SiO2/SnO2 mixed oxides obtained by the sol-gel process. Evaluation of immobilized copper hexacyanoferrate as an electrochemical probe

SiO2/SnO2 mixed oxides were prepared by the sol-gel processing method using SnI4 as the tin oxide precursor reagent. Solids with Sri compositions (in wt.%) of 4.1, 12.9 and 18.4 and presenting specific surface areas (detennined by the BET method) of 492, 658 and 712 m 2 (-l) 9, respectively, were obtained. Transmission microscopic images showed nanosized SnO2 particles with average dimensions of (5.3 +/- 0.5) mu for samples having 12.9 wt.% Sri and (7.0 +/- 0.7) nm for samples having 18.4 wt.% Sri. For the sample presenting 4 wt.% of Sn the crystallites were poorly defined, barely being observed. The amorphous SnO2 particles started to crystallize at different temperatures, i.e., 1273, 1173 and 1073 K for samples with 4.1, 12.9 and 18.4 wt.% of Sri, respectively. The X-ray diffraction patterns showed that only cassiterite crystallites were present in every case and, even at a temperature of 1473 K, the SiO2, remained as an amorphous matrix. The Lewis and Bronsted acid sites were thermally stable up to a temperature of 523 K for all the compositions, as probed using pyridine molecules. The infrared spectra showed that Si-O-Sn bonds are formed at the interface between SiO2 and SnO2 particles. These bonds are the ones mainly responsible for the low mobility of the oxide particles, avoiding crystallization of SnO2. Copper hexacyanoferrate was prepared in situ on the SiO2/SnO2 surface and cyclic voltammetry tests were carried out by sweeping the potential between 0.2 and 1.0 V. The midpoint potential corresponding to the redo process: equivalent to SnOH2+/{KCu potential between 0.2 and 1.0 V. The midpoint potential corresponding to the redox process: SnOH2+/KCu[Fe(CN)(6)}(-) Cu[Fe(CN)(6)}(_)+K++e(-) was observed at about 0.7 V. The electrochemical impedance spectroscopic data showed a charge transfer resistance of 17.8 Omega. This low value favors the oxidation-reduction process in the pores of the material. (C) 2004 Elsevier B.V All rights reserved.1674167116517

Clinical characteristics of small functioning adrenocortical tumors in children

Twenty of 67 children registered on the International Registry of Childhood Adrenocortical Tumors between May 1988 and December 1994 had small adrenocortical tumors (defined for this study as measuring less than or equal to 200 cm(3) and/or weighing less than or equal to 100 g). We reviewed the records of these 20 patients to characterize the clinical and pathologic findings and outcomes of children with small adrenocortical tumors. Median patient age was 2 years (range, 4 months to 5 years). There was only one boy. All had clinical signs of virilization, and seven had signs or symptoms of Cushing syndrome. A median 5.5 months (range, 1-40 months) had elapsed between the first signs of endocrine dysfunction and diagnosis. All tumors were surgically resected. Tumor volume was 3.3-195 cm(3) (median, 38.7 cm(3)), and weight was 3.7-100 g (median, 36 g). Tumor samples were histologically reviewed in 18 cases. Eight were adenomas, and 10 were carcinomas (6 low grade and 4 high grade). Pathology records described tumor with diagnostic features of adrenocortical carcinoma in two patients. One patient received mitotane for 8 months after surgery. Only one patient had recurrent disease, which was detected 6 months after diagnosis and proved rapidly fatal. Another has been lost to follow-up. the remaining 18 patients are alive with no evidence of disease at a median 2.3 years (range, 6 months to 6.1 years) after diagnosis. Our data suggest that children with small adrenocortical tumors have an excellent prognosis with surgery as the sole therapy regardless of tumor histiotype. (C) 1997 Wiley-Liss, Inc.ST JUDE CHILDRENS RES HOSP, DEPT HEMATOL ONCOL, RCR, MEMPHIS, TN 38105 USAST JUDE CHILDRENS RES HOSP, DEPT SURG, MEMPHIS, TN 38105 USAST JUDE CHILDRENS RES HOSP, INT OUTREACH PROGRAM, MEMPHIS, TN 38105 USAUNIV FED PARANA, DEPT PEDIAT, BR-80060000 CURITIBA, PARANA, BRAZILUNIV São Paulo, BR-05508 São Paulo, BRAZILESCOLA PAULISTA MED, São Paulo, BRAZILUNIV TENNESSEE, DEPT PATHOL, MEMPHIS, TN USAUNIV TENNESSEE, DEPT PEDIAT, MEMPHIS, TN USACOLL MED, MEMPHIS, TN USAESCOLA PAULISTA MED, São Paulo, BRAZILWeb of Scienc