Extracting histones for the specific purpose of label-free MS

Extracting histones from cells is the first step in studies that aim to characterize histones and their post-translational modifications (hPTMs) with MS. In the last decade, label-free quantification is more frequently being used for MS-based histone characterization. However, many histone extraction protocols were not specifically designed for label-free MS. While label-free quantification has its advantages, it is also very susceptible to technical variation. Here, we adjust an established histone extraction protocol according to general label-free MS guidelines with a specific focus on minimizing sample handling. These protocols are first evaluated using SDS-PAGE. Hereafter, a selection of extraction protocols was used in a complete histone workflow for label-free MS. All protocols display nearly identical relative quantification of hPTMs. We thus show that, depending on the cell type under investigation and at the cost of some additional contaminating proteins, minimizing sample handling can be done during histone isolation. This allows analyzing bigger sample batches, leads to reduced technical variation and minimizes the chance of in vitro alterations to the hPTM snapshot. Overall, these results allow researchers to determine the best protocol depending on the resources and goal of their specific study. Data are available via ProteomeXchange with identifier PXD002885

Rupture of Splenic Artery Aneurysm in Patient with ACTN2 Mutation

Here, we report a case of splenic artery aneurysm rupture in a patient with known heterozygosity mutation of the ACTN2 gene (variant c.971G > A p.Arg324Gln). The patient came to our emergency department with epigastric pain radiating to the lumbar area, with an absence of peritonism signs. An abdominal computed tomography angiography showed a ruptured huge (5 cm) splenic artery aneurysm. Therefore, the patient underwent emergency endovascular coil embolization with complete aneurysm exclusion. The postoperative course was uneventful, until postoperative day five when the patient developed a symptomatic supraventricular tachycardia in the absence of echocardiographic alterations. The signs and symptoms disappeared after three days of medical management. The patient was discharged on the 14th postoperative day in good clinical condition under verapamil and anti-platelet therapy. Although ACTN2 mutation was associated with cardiac and peripheral vascular disease occurrence, to the best of our knowledge, the present case is the first report of a visceral (splenic) aneurysm directly linked with this rare mutation

Simple, Exact Density-Functional-Theory Embedding Scheme

Density functional theory (DFT) provides a formally exact framework for quantum embedding. The appearance of nonadditive kinetic energy contributions in this context poses significant challenges, but using optimized effective potential (OEP) methods, various groups have devised DFT-in-DFT methods that are equivalent to Kohn–Sham (KS) theory on the whole system. This being the case, we note that a very considerable simplification arises from doing KS theory instead. We then describe embedding schemes that enforce Pauli exclusion via a projection technique, completely avoiding numerically demanding OEP calculations. Illustrative applications are presented using DFT-in-DFT, wave-function-in-DFT, and wave-function-in-Hartree–Fock embedding, and using an embedded many-body expansion

Oscillation of plantar pressure center in athletes and non-athletes with and without ankle sprains

AbstractObjectiveTo assess whether there is any difference in the oscillation of the plantar pressure center in single-leg stance between athletes and non-athletes with and without ankle sprains.Methods54 volunteers performed four static assessments and one dynamic assessment while standing on one foot on a baropodometer, barefoot, for 10s in each test. The variables of area (cm2), distance (cm), anteroposterior oscillation (cm), mediolateral oscillation (cm) and mean velocity (cm/s) were analyzed. The items “other symptoms” and “sports and recreation” of the subjective Foot and Ankle Outcome Score (FAOS) questionnaire were applied. For the statistical analysis, repeated-measurement ANOVA (ANOVA-MR), multivariate ANOVA (MANOVA), Tukey's post hoc test and partial eta squared were used.ResultsANOVA-MR revealed differences regarding distance, with major effects for eyes (p<0.001), knees (p<0.001), group (p<0.05) and the interaction between eyes and knees (p<0.05); and regarding mean velocity with major effects for eyes (p<0.001), knees (p<0.001) (p<0.05), group (p<0.05) and the interaction between eyes and knees (p<0.05). MANOVA revealed main group effects for distance (p<0.05), anteroposterior oscillation (p<0.05) and mean velocity (p<0.05). In the FAOS questionnaire, there were no differences: “other symptoms”, p>0.05; and “sport and recreation”, p>0.05.ConclusionAthletes present higher mean velocity of oscillation of plantar pressure center and generally do not have differences in oscillation amplitude in the sagittal and coronal planes, in comparison with non-athletes

Genomic, transcriptomic and RNA editing analysis of human MM1 and VV2 sporadic Creutzfeldt-Jakob disease

Creutzfeldt-Jakob disease (CJD) is characterized by a broad phenotypic spectrum regarding symptoms, progression, and molecular features. Current sporadic CJD (sCJD) classification recognizes six main clinical-pathological phenotypes. This work investigates the molecular basis of the phenotypic heterogeneity of prion diseases through a multiomics analysis of the two most common sCJD subtypes: MM1 and VV2. We performed DNA target sequencing on 118 genes on a cohort of 48 CJD patients and full exome RNA sequencing on post-mortem frontal cortex tissue on a subset of this cohort. DNA target sequencing identified multiple potential genetic contributors to the disease onset and phenotype, both in terms of coding, damaging-predicted variants, and enriched groups of SNPs in the whole cohort and the two subtypes. The results highlight a different functional impairment, with VV2 associated with higher impairment of the pathways related to dopamine secretion, regulation of calcium release and GABA signaling, showing some similarities with Parkinson’s disease both on a genomic and a transcriptomic level. MM1 showed a gene expression profile with several traits shared with different neurodegenerative, without an apparent distinctive characteristic or similarities with a specific disease. In addition, integrating genomic and transcriptomic data led to the discovery of several sites of ADAR-mediated RNA editing events, confirming and expanding previous findings in animal models. On the transcriptomic level, this work represents the first application of RNA sequencing on CJD human brain samples. Here, a good clusterization of the transcriptomic profiles of the two subtypes was achieved, together with the finding of several differently impaired pathways between the two subtypes. The results add to the understanding of the molecular features associated with sporadic CJD and its most common subtypes, revealing strain-specific genetic signatures and functional similarities between VV2 and Parkinson’s disease and providing preliminary evidence of RNA editing modifications in human sCJD

The relationship between knowing sign language and quality of life among Italian people who are deaf : a cross-sectional study

Deafness is a medical condition with important relational implications. This condition could affect well-being and self-esteem and cause social anxiety. Sign language is not only a simple mimic but can be considered as a different kind of communication that could be protective for those who have learned it. However, some people do not use sign language because they think it can be marginalizing. The present study aimed to compare the quality of life (QoL) between people who learned Italian sign language as their first language with those who had never learned it or learned it later. This cross-sectional study involved 182 deaf Italian adults (70.3% females) who were recruited from Ente Nazionale Sordi (ENS) and by the main online deafness groups. The present results suggest that the deaf condition does not seem to significantly affect the dimensions of QoL pertaining to satisfaction and self-esteem, while it could have an effect on preventing high levels of social anxiety and in particular, the group who learned Italian sign language showed significantlyless social anxiety than those who had never learned it

Dietary methionine source alters the lipidome in the small intestinal epithelium of pigs

Methionine (Met) as an essential amino acid has key importance in a variety of metabolic pathways. This study investigated the influence of three dietary Met supplements (0.21% L-Met, 0.21% DL-Met and 0.31% DL-2-hydroxy-4-(methylthio)butanoic acid (DL-HMTBA)) on the metabolome and inflammatory status in the small intestine of pigs. Epithelia from duodenum, proximal jejunum, middle jejunum and ileum were subjected to metabolomics analysis and qRT-PCR of caspase 1, NLR family pyrin domain containing 3 (NLRP3), interleukins IL1β, IL8, IL18, and transforming growth factor TGFβ. Principal component analysis of the intraepithelial metabolome revealed strong clustering of samples by intestinal segment but not by dietary treatment. However, pathway enrichment analysis revealed that after L-Met supplementation polyunsaturated fatty acids (PUFA) and tocopherol metabolites were lower across small intestinal segments, whereas monohydroxy fatty acids were increased in distal small intestine. Pigs supplemented with DL-HMTBA showed a pronounced shift of secondary bile acids (BA) and sphingosine metabolites from middle jejunum to ileum. In the amino acid super pathway, only histidine metabolism tended to be altered in DL-Met-supplemented pigs. Diet did not affect the expression of inflammation-related genes. These findings suggest that dietary supplementation of young pigs with different Met sources selectively alters lipid metabolism without consequences for inflammatory status