Sex-specific pleiotropic changes in emotional behavior and alcohol consumption in human α-synuclein A53T transgenic mice during early adulthood.

Point mutations in the α-synuclein coding gene may lead to the development of Parkinson's disease (PD). PD is often accompanied by other psychiatric conditions, such as anxiety, depression, and drug use disorders, which typically emerge in adulthood. Some of these point mutations, such as SNCA and A30T, have been linked to behavioral effects that are not commonly associated with PD, especially regarding alcohol consumption patterns. In this study, we investigated whether the familial PD point mutation A53T is associated with changes in alcohol consumption behavior and emotional states at ages not yet characterized by α-synuclein accumulation. The affective and alcohol-drinking phenotypes remained unaltered in female PDGF-hA53T-synuclein-transgenic (A53T) mice during both early and late adulthood. Brain region-specific activation of ceramide-producing enzymes, acid sphingomyelinase (ASM), and neutral sphingomyelinase (NSM), known for their neuroprotective properties, was observed during early adulthood but not in late adulthood. In males, the A53T mutation was linked to a reduction in alcohol consumption in both early and late adulthood. However, male A53T mice displayed increased anxiety- and depression-like behaviors during both early and late adulthood. Enhanced ASM activity in the dorsal mesencephalon and ventral hippocampus may potentially contribute to these adverse behavioral effects of the mutation in males during late adulthood. In summary, the A53T gene mutation was associated with diverse changes in emotional states and alcohol consumption behavior long before the onset of PD, and these effects varied by sex. These alterations in behavior may be linked to changes in brain ceramide metabolism.

An ensemble-based feature selection framework to select risk factors of childhood obesity for policy decision making.

BACKGROUND: The increasing prevalence of childhood obesity makes it essential to study the risk factors with a sample representative of the population covering more health topics for better preventive policies and interventions. It is aimed to develop an ensemble feature selection framework for large-scale data to identify risk factors of childhood obesity with good interpretability and clinical relevance. METHODS: We analyzed the data collected from 426,813 children under 18 during 2000-2019. A BMI above the 90th percentile for the children of the same age and gender was defined as overweight. An ensemble feature selection framework, Bagging-based Feature Selection framework integrating MapReduce (BFSMR), was proposed to identify risk factors. The framework comprises 5 models (filter with mutual information/SVM-RFE/Lasso/Ridge/Random Forest) from filter, wrapper, and embedded feature selection methods. Each feature selection model identified 10 variables based on variable importance. Considering accuracy, F-score, and model characteristics, the models were classified into 3 levels with different weights: Lasso/Ridge, Filter/SVM-RFE, and Random Forest. The voting strategy was applied to aggregate the selected features, with both feature weights and model weights taken into consideration. We compared our voting strategy with another two for selecting top-ranked features in terms of 6 dimensions of interpretability. RESULTS: Our method performed the best to select the features with good interpretability and clinical relevance. The top 10 features selected by BFSMR are age, sex, birth year, breastfeeding type, smoking habit and diet-related knowledge of both children and mothers, exercise, and Mother's systolic blood pressure. CONCLUSION: Our framework provides a solution for identifying a diverse and interpretable feature set without model bias from large-scale data, which can help identify risk factors of childhood obesity and potentially some other diseases for future interventions or policies.

Leiomioma celular gigante con degeneración roja, hidrópica y quística: reporte de un caso / Giant cellular leiomioma with red, hydropic and cystic degeneration: report of a case

Los miomas uterinos son neoplasias bastante comunes, se clasifican según su localización en el miometrio; subseroso, intramural, transmural y submucoso. A su vez existen diferentes variantes morfológicas y cambios degenerativos que son un fenómeno frecuente en los miomas, especialmente entre mayor tamaño tenga éste: como la apoplejía, atrofia, hialinización, degeneración roja, quística, calcificación periférica, necrosis, degeneración hidrópica y la degeneración maligna o sarcomatosa. Se expone el caso de una paciente de 53 años sometida a una laparotomía donde se identificó una masa tumoral de 8150 gramos, que dio como resultado anatomopatológico un leiomioma gigante con 3 diversos tipos de degeneración: roja, hidrópica y quística. Se informa este caso por la rareza en su conformación y debido a la falta de reportes en la literatura mundial, siendo este el primer caso presentando y permitiendo realizar algunos comentarios sobre la patología.

Uterine myomas are quite common neoplasms. They are classified according to their location in the myometrium; subserosa, intramural, transmural and submucosal; At the same time, there are different morphological variants and degenerative changes that are a frequent phenomenon in myomas, especially the larger it is: as the apoplexy, atrophy, hyalinization, red degeneration, cystic, peripheral calcification, necrosis, hydropic degeneration and malignant degeneration or sarcomatous. We present the case of a 53-year-old patient who underwent a laparotomy where a tumor mass of 8150 grams was identified, which resulted in a pathological result of a giant leiomyoma with 3 different types of degeneration: red, hydropic and cystic. This case is reported due to the rarity of its conformation and due to the lack of reports in the world literature, this being the first case presenting and allowing some comments on the pathology.

Different EV enrichment methods suitable for clinical settings yield different subpopulations of urinary extracellular vesicles from human samples.

Urine sample analysis is irreplaceable as a non-invasive method for disease diagnosis and follow-up. However, in urine samples, non-degraded protein and RNA may be only found in urinary extracellular vesicles (uEVs). In recent years, various methods of uEV enrichment using low volumes of urine and unsophisticated equipment have been developed, with variable success. We compared the results of the differential ultracentrifugation procedure with 4 of these methods. The methods tested were a lectin-based purification, Exoquick (System Biosciences), Total Exosome Isolation from Invitrogen and an in-house modified procedure employing the Exosomal RNA Kit from Norgen Biotek Corp. The analysis of selected gene transcripts and protein markers of extracellular vesicles (EVs) revealed that each method isolates a different mixture of uEV protein markers. In our conditions, the extraction with Norgen's reagent achieved the best performance in terms of gene transcript and protein detection and reproducibility. By using this method, we were able to detect alterations of EVs protein markers in urine samples from prostate cancer adenoma patients. Taken together, our results show that the isolation of uEVs is feasible from small volumes of urine and avoiding ultracentrifugation, making easier the analysis in a clinical facility. However, caution should be taken in the selection of the enrichment method since they have a differential affinity for protein uEVs markers and by extension for different subpopulation of EVs.

Methodological aspects of the molecular and histological study of prostate cancer: focus on PTEN.

Prostate cancer is among the most frequent cancers in men, and despite its high rate of cure, the high number of cases results in an elevated mortality worldwide. Importantly, prostate cancer incidence is dramatically increasing in western societies in the past decades, suggesting that this type of tumor is exquisitely sensitive to lifestyle changes. Prostate cancer frequently exhibits alterations in the PTEN gene (inactivating mutations or gene deletions) or at the protein level (reduced protein expression or altered sub-cellular compartmentalization). The relevance of PTEN in this type of cancer is further supported by the fact that the sole deletion of PTEN in the murine prostate epithelium recapitulates many of the features of the human disease. In order to study the molecular alterations in prostate cancer, we need to overcome the methodological challenges that this tissue imposes. In this review we present protocols and methods, using PTEN as proof of concept, to study different molecular characteristics of prostate cancer.

Sources of pre-analytical variations in yield of DNA extracted from blood samples: analysis of 50,000 DNA samples in EPIC.

The European Prospective Investigation into Cancer and nutrition (EPIC) is a long-term, multi-centric prospective study in Europe investigating the relationships between cancer and nutrition. This study has served as a basis for a number of Genome-Wide Association Studies (GWAS) and other types of genetic analyses. Over a period of 5 years, 52,256 EPIC DNA samples have been extracted using an automated DNA extraction platform. Here we have evaluated the pre-analytical factors affecting DNA yield, including anthropometric, epidemiological and technical factors such as center of subject recruitment, age, gender, body-mass index, disease case or control status, tobacco consumption, number of aliquots of buffy coat used for DNA extraction, extraction machine or procedure, DNA quantification method, degree of haemolysis and variations in the timing of sample processing. We show that the largest significant variations in DNA yield were observed with degree of haemolysis and with center of subject recruitment. Age, gender, body-mass index, cancer case or control status and tobacco consumption also significantly impacted DNA yield. Feedback from laboratories which have analyzed DNA with different SNP genotyping technologies demonstrate that the vast majority of samples (approximately 88%) performed adequately in different types of assays. To our knowledge this study is the largest to date to evaluate the sources of pre-analytical variations in DNA extracted from peripheral leucocytes. The results provide a strong evidence-based rationale for standardized recommendations on blood collection and processing protocols for large-scale genetic studies.

High-density SNP genotyping detects homogeneity of Spanish and French Basques, and confirms their genomic distinctiveness from other European populations.

A recent study reported that Basques do not constitute a genetically distinct population, and that Basques from Spanish and French provinces do not show significant genetic similarity. These conclusions disagree with numerous previous studies, and are not consistent with the historical and linguistic evidence that supports the distinctiveness of Basques. In order to further investigate this controversy, we have genotyped 83 Spanish Basque individuals and used these data to infer population structure based on more than 60,000 single nucleotide polymorphisms of several European populations. Here, we present the first high-throughput analysis including Basques from Spanish and French provinces, and show that all Basques constitute a homogeneous group that can be clearly differentiated from other European populations.

Polymorphisms in metabolic genes related to tobacco smoke and the risk of gastric cancer in the European prospective investigation into cancer and nutrition.

Metabolizing enzymes, which often display genetic polymorphisms, are involved in the activation of compounds present in tobacco smoke that may be relevant to gastric carcinogenesis. We report the results of a study looking at the association between risk of gastric adenocarcinoma and polymorphisms in genes CYP1A1, CYP1A2, EPHX1, and GSTT1. A nested case-control study was carried out within the European Prospective Investigation into Cancer and Nutrition, developed in 10 European countries. The study includes 243 newly diagnosed cases of histologically confirmed gastric adenocarcinoma and 946 controls matched by center, age, sex, and date of blood collection. Genotypes were determined in nuclear DNA from WBCs. We found an increased risk of gastric cancer for homozygotes for C (histidine) variant in Y113H of EPHX1 (odds ratio, 1.91; 95% confidence interval, 1.19-3.07) compared with subjects with TC/TT. There was also a significant increased risk for smokers carrying at least one variant allele A in Ex7+129C>A (m4) of CYP1A1 and never smokers with null GSTT1 and allele A in the locus -3859G>A of CYP1A2. Most of these genes are involved in the activation and detoxification of polycyclic aromatic hydrocarbons, suggesting a potential role of these compounds in gastric carcinogenesis.

Patterns of gene expression from in utero delivery of adenoviral-associated vector serotype 1.

Adenoviral-associated viral vectors (AAV) have shown significant promise for efficient gene delivery to multiple tissues. Studies of different serotypes of AAV revealed different expression patterns provided by gene delivery in postnatal mice. Previous in utero gene delivery studies of AAV serotype 2 (AAV2) demonstrated efficient gene expression in certain fetal tissues depending on route of administration. We studied the pattern of gene expression from AAV serotype 1 (AAV1) using intramuscular, intraperitoneal, and intravascular routes of administration in embryonic day 16 C57BL/6 mice. Limb skeletal muscle transduction was only achieved with AAV1 by intramuscular administration. The levels of gene expression were 20-fold higher than a comparable administration of AAV2. Diaphragm muscle transduction by AAV1 was achieved at the highest level by intraperitoneal administration, and to a lesser degree by intravascular administration. All delivery routes resulted in transgene expression in the lung. Our results indicate that AAV1 can offer higher transgene expression in fetal skeletal muscle than AAV2 with intramuscular administration. The transgene expression pattern in different tissues, which depends on vector serotype and route of administration, will need to be considered in planning therapeutic studies for specific disorders.

Biocompatibility of adenoviral vectors in poly(vinyl chloride) tubing catheters with presence or absence of plasticizer di-2-ethylhexyl phthalate.

Adenoviral (Ad) vectors feature attractive characteristics for gene therapy of a wide variety of diseases. In many cases, the Ad vector must be administered using catheters and other plastic medical devices. Although poly(vinyl chloride) is one of the most frequently used catheter materials, it is relatively rigid and requires the addition of a plasticizer such as di-2-ethylhexyl phthalate (DEHP) to increase its flexibility. In this study, we demonstrated that exposure to a DEHP-containing catheter decreased the infectivity of Ad vectors but not the total particle number of the vector. Loss of Ad vector infectivity was directly related to the time of exposure to the DEHP-containing catheter, but it was not due to simple leaching of the chemical from the plastic. The loss of Ad vector infectivity could be prevented by preflushing the tube with albumin. Careful consideration of the compatibility between gene therapy vectors and medical delivery devices will be critical to the success of human gene therapy applications.

Binding of adenoviral fiber knob to the coxsackievirus-adenovirus receptor is crucial for transduction of fetal muscle.

Adenoviral (Ad) infection involves attachment mediated by the Ad fiber protein binding to the coxsackievirus-adenovirus receptor (CAR) of a target cell and internalization facilitated by the interaction of the Ad penton base protein with alpha(v) integrins. To understand the relative importance of the Ad binding and internalization steps for the transduction of fetal skeletal muscle, we used a panel of genetically modified vectors that specifically ablate the fiber-CAR interaction (AdL.F*), the penton base-alpha(v) integrin interaction (AdL.PB*), or both (AdL.PB*F*) to transduce embryonic day 16 (E-16) mouse muscle in vivo and primary E-16 muscle cells in vitro. Quantification of transgene expression and vector genome copies revealed a striking absence of E-16 muscle transduction by AdL.F* and AdL.PB*F*. In contrast, fetal muscle transduction with AdL.PB* was not significantly different than with the unmodified vector. Similar results were observed with in vitro Ad infection studies in primary E-16 muscle cells. From these data we conclude that the fiber-CAR interaction is important for the transduction of fetal muscle by Ad vectors. The high dependence on fiber-CAR binding will impact the development of strategies for Ad vector retargeting to achieve muscle-specific transduction in utero.