Regulation of Redox Profile and Genomic Instability by Physical Exercise Contributes to Neuroprotection in Mice with Experimental Glioblastoma.

Glioblastoma (GBM) is an aggressive, common brain cancer known to disrupt redox biology, affecting behavior and DNA integrity. Past research remains inconclusive. To further understand this, an investigation was conducted on physical training's effects on behavior, redox balance, and genomic stability in GBMA models. Forty-seven male C57BL/6J mice, 60 days old, were divided into GBM and sham groups (n = 15, n = 10, respectively), which were further subdivided into trained (Str, Gtr; n = 10, n = 12) and untrained (Sut, Gut; n = 10, n = 15) subsets. The trained mice performed moderate aerobic exercises on a treadmill five to six times a week for a month while untrained mice remained in their enclosures. Behavior was evaluated using open-field and rotarod tests. Post training, the mice were euthanized and brain, liver, bone marrow, and blood samples were analyzed for redox and genomic instability markers. The results indicated increased latency values in the trained GBM (Gtr) group, suggesting a beneficial impact of exercise. Elevated reactive oxygen species in the parietal tissue of untrained GBM mice (Gut) were reduced post training. Moreover, Gtr mice exhibited lower tail intensity, indicating less genomic instability. Thus, exercise could serve as a promising supplemental GBM treatment, modulating redox parameters and reducing genomic instability.

Functional Analysis of LDLR (Low-Density Lipoprotein Receptor) Variants in Patient Lymphocytes to Assess the Effect of Evinacumab in Homozygous Familial Hypercholesterolemia Patients With a Spectrum of LDLR Activity.

OBJECTIVE: Homozygous familial hypercholesterolemia is a rare disease usually caused by LDLR (low-density lipoprotein receptor) mutations. Homozygous familial hypercholesterolemia is characterized by markedly elevated LDL-C (low-density lipoprotein cholesterol) levels and an extremely high risk of premature atherosclerotic cardiovascular disease. A phase 2, proof-of-concept study (NCT02265952) demonstrated that evinacumab, a fully human monoclonal antibody to ANGPTL3 (angiopoietin-like 3 protein), reduced LDL-C levels in 9 patients with genotypically confirmed homozygous familial hypercholesterolemia and was well tolerated. The aim of this study was to analyze the effects of evinacumab on LDLR activity in lymphocytes purified from patients in the proof-of-concept study. Approach and Results: LDLR activity was assessed in patient lymphocytes before and after treatment with evinacumab and versus lymphocytes carrying wild-type LDLR, and also in an LDLR-defective Chinese hamster ovary cell line (CHO-ldlA7) transfected with plasmids encoding the LDLR variants. Overall mean peak reduction in LDL-C with evinacumab was -58±18%, occurring between Week 4 and Week 12. Mutations identified in the 9 patients were shown to be pathogenic, with loss of LDLR activity versus wild type. Two of the LDLR variants, p.(Cys681*) and p.(Ala627Profs*38), were class 2 type mutations that are retained in the endoplasmic reticulum. Six variants were class 3 type mutations with impaired LDL-C binding activity: p.(Trp87Gly), occurring in 2 patients, p.(Gln254Pro), p.(Ser177Leu), p.(Gly335Val), and p.(Ser306Leu). Evinacumab had no effect on LDLR activity. CONCLUSIONS: These results suggest that evinacumab is effective for lowering LDL-C in patients with homozygous familial hypercholesterolemia, and the inhibition of ANGPTL3 in humans lowers LDL-C in a mechanism independent of the LDLR.

Low-density lipoprotein receptor mutational analysis in diagnosis of familial hypercholesterolemia.

PURPOSE OF REVIEW: To present up to date evidence on the pathogenicity of low-density lipoprotein receptor (LDLR) variants and to propose a strategy that is suitable for implementation in the clinical work-up of familial hypercholesterolaemia. RECENT FINDINGS: More than 1800 variants have been described in the LDLR gene of patients with a clinical diagnosis of familial hypercholesterolaemia; however, less than 15% have functional evidence of pathogenicity. SUMMARY: The spectrum of variants in the LDLR identified in patients with clinical familial hypercholesterolaemia is increasing as novel variants are still being reported. However, over 50% of all LDLR variants need further evidence before they can be confirmed as mutations causing disease. Even with applying the recent American College of Medical Genetics variant classification, a large number of variants are still considered variants of unknown significance. Before obtaining an undisputable confirmation of the effect on the expression and activity of the LDLR, reporting these variants as part of a clinical diagnosis to the patient holds the risk that it might need to be withdrawn in a later stage. An investment should be made to develop functional assays to characterize LDLR variants of unknown significance for a better patient diagnosis and to prevent confusion in the physician's office.

Descrição morfológica do coração e dos vasos da base do jacaré-do-pantanal (Caiman yacare Daudin, 1802) proveniente de zoocriadouro / Morphological description of heart and large vessels of the jacaré-do-pantanal (Caiman yacare Daudin, 1802) from a crocodile breeding Center

Com este estudo objetivou-se descrever os aspectos anatômicos e histológicos do coração do jacaré-do-pantanal (Caiman yacare), proveniente de zoocriadouro. Para tanto, estudou-se 13 exemplares da espécie, os quais foram perfundidos, conservados em solução de formaldeído a 10% e submetidos às técnicas anatômicas específicas. O coração foi separado e amostras foram colhidas e submetidas à avaliação histológica. Macroscopicamente o coração é tetracavitário, e além de dois átrios e dois ventrículos, apresenta uma estrutura denominada cone arterial, do qual emergem os vasos da base do coração. Foram identificadas duas aortas, direita e esquerda, sendo que a esquerda emerge do ventrículo direito e se comunica com o tronco sistêmico direito por meio do forame de Panizza. Histologicamente o coração possui epicárdio, miocárdio e endocárdio típicos. Concluímos que a histologia do coração, no jacaré-do-pantanal, é semelhante à de outras espécies de répteis. Contudo, anatomicamente apresenta particularidades importantes, as quais representam, possivelmente, adaptações que permitiram a perpetuação da espécie.(AU)

The aim of this study was to describe anatomical and histological aspects of the heart of Caiman yacare from a crocodile breeding center. For this purpose, we have chosen and further studied 13 specimens which were perfused and preserved in a 10% formaldehyde solution and they were subjected to the specific anatomical techniques. The heart was separated and samples were collected and submitted to the specific histological procedures. Macroscopically, the heart is four-chambered and besides two atria and two ventricles, has a structure called arterial cone from which the large vessels of the heart emerge. Two aortas, left and right, were identified. The left aorta emerges from the right ventricle and communicates with the systemic trunk through the Foramen of Panizza. From a histological point of view, the heart is typically composed of epicardium, myocardium and endocardium. According to these observations, it is assumed that the histology of the heart of "Jacaré-do-Pantanal" is similar to other species of reptiles. However, there are some anatomic particularities, which possibly represent the adaptations allowing the perpetuation of the species.(AU)

Advantages and versatility of fluorescence-based methodology to characterize the functionality of LDLR and class mutation assignment.

Familial hypercholesterolemia (FH) is a common autosomal codominant disease with a frequency of 1:500 individuals in its heterozygous form. The genetic basis of FH is most commonly mutations within the LDLR gene. Assessing the pathogenicity of LDLR variants is particularly important to give a patient a definitive diagnosis of FH. Current studies of LDLR activity ex vivo are based on the analysis of 125I-labeled lipoproteins (reference method) or fluorescent-labelled LDL. The main purpose of this study was to compare the effectiveness of these two methods to assess LDLR functionality in order to validate a functional assay to analyse LDLR mutations. LDLR activity of different variants has been studied by flow cytometry using FITC-labelled LDL and compared with studies performed previously with 125I-labeled lipoproteins. Flow cytometry results are in full agreement with the data obtained by the 125I methodology. Additionally confocal microscopy allowed the assignment of different class mutation to the variants assayed. Use of fluorescence yielded similar results than 125I-labeled lipoproteins concerning LDLR activity determination, and also allows class mutation classification. The use of FITC-labelled LDL is easier in handling and disposal, cheaper than radioactivity and can be routinely performed by any group doing LDLR functional validations.

Qualidade do leite de vacas criadas no sistema silvipastoril no Vale do Cauca, Colômbia / Milk quality indicators from cows bred on silvopastoral system in Cauca Valley, Colombia

O sistema silvipastoril caracteriza-se por aumentar a produção de leite, com maior número de vacas por hectare devido ao maior aporte de proteína na dieta. Neste sistema as vacas são alimentadas, além do pasto, de pequenas árvores e arbustos. O objetivo do presente estudo foi determinar os principais indicadores de qualidade do leite e agentes causais de mastite em vacas criadas em sistema silvipastoril. Foram avaliadas a composição (teor de gordura, proteína total, lactose, extrato seco, extrato seco desengordurado e nitrogênio uréico), contagem de células somáticas (CCS), contagem bacteriana total (CBT), ocorrência de mastite clínica e subclínica, isolamento microbiológico, perfil de sensibilidade bacteriana "in vitro" e detecção de resíduos antimicrobianos no leite produzido por 100 vacas, bem como do tanque de expansão e latões em propriedades do Vale do Cauca, Colômbia. Os teores médios dos principais constituintes do leite foram 3,24% de gordura, 3,27% de proteína total, 4,40% de lactose, 10,62% de extrato seco, 8,57% de extrato seco desengordurado e 15,82mg/dL de nitrogênio uréico, enquanto do tanque de expansão e latões foi 3,51% de gordura, 3,20% de proteína total, 4,34% de lactose, 11,72% de extrato seco, 8,47% de extrato seco desengordurado e 14,57mg/dL de nitrogênio uréico. A celularidade média dos quartos mamários e do tanque de expansão foi 141.252,75 CS/mL e 363.078,05 CS/mL respectivamente. A CBT média dos quartos mamários e do tanque de expansão foi 4.466,84 UFC/mL e 24.547,01 UFC/mL. Os principais micro-organismos isolados dos quartos mamários foram Corynebacterium bovis, Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus spp., Streptococcus dysgalatiae, enquanto do tanque de expansão foram identificados Streptococcus spp., Enterobacter cloacae, Hafnia alveii, Streptococcus hemolítico e Streptococcus spp., com maior frequência. A presença de resíduos de antimicrobianos em leite de vacas e do tanque ou latão foi detectada em 30% e 86% das amostras, respectivamente. O sistema silvipastoril mostrou ser uma boa alternativa para produção de leite em vacas. No entanto, são necessários cuidados no tratamento mamário para evitar resíduos no leite e a análise de todos os parâmetros de qualidade para garantir um produto diferenciado.

The silvopastoral system is characterized by increasing the production of milk with a greater number of cows per hectare due to the higher amount of protein in the diet. In silvopastoral system cows are fed in addition to pasture, small trees and shrubs. The aim of this study was determinate the main indicators of milk quality and mastitis causal agents in cows bred on silvopastoral system. We evaluated the composition (fat, total protein, lactose, solids, dry extract, nonfat dry and urea nitrogen), somatic cell count (SCC), total bacterial count (TBC), occurrence of clinical and subclinical mastitis, microbiological isolation, in vitro antimicrobial sensitivity profile and detection of antimicrobial residues in milk produced by 100 cows raised in silvopastoral systems, as well as the bulk tank and churns in farms of Cauca Valley, Colombia. The concentration of the major constituents of milk were 3.24% fat, 3.27% total protein, 4.40% lactose, 10.62% dry extract, 8.57% nonfat dry and 15.82mg/dL urea nitrogen, while the bulk tank and churns was 3.51% fat, 3.20% total protein, 4.34% lactose, 11.72% dry extract, 8.47% nonfat dry and 14.57mg/dL urea nitrogen. The cell count of the cows and the bulk tank was 141,252.75 CS/mL and 363,078.05 CS/mL respectively. The TBC mean in cows and the bulk tank was 4,466.84 CFU/mL and 24,547.01 CFU/mL respectively. The main microorganisms isolated from the udder cows were Corynebacterium bovis, Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus spp., Streptococcus dysgalatiae, while the bulk tank were identified more often Streptococcus spp., Enterobacter cloacae, Hafnia alveii, hemolytic Streptococcus and Streptococcus spp. Antimicrobial residues in cow milk and bulk or churn were detected in 30% and 86% respectively. The silvopastoral system showed to be good alternative to milk production in cow. However is important the care with antimicrobial residues in milk and the analysis of all quality parameters to ensure a differentiated product.

How good is controlled attenuation parameter and fatty liver index for assessing liver steatosis in general population: correlation with ultrasound.

BACKGROUND & AIMS: Liver steatosis measurement by controlled attenuation parameter (CAP) is a non-invasive method for diagnosing steatosis, based on transient elastography. Its usefulness as screening procedure for hepatic steatosis in general population has not been previously evaluated. The aim of this study was to evaluate the diagnostic accuracy of CAP and fatty liver index (FLI) for detection and quantification of steatosis in general population. METHODS: Recruitment was done from a prospective epidemiological study of the general adult population. Steatosis was evaluated using CAP, FLI and ultrasound (US). Steatosis scored according to Hamaguchi's US scoring, from 0 (S0) to 6 (S6) points. Hepatic steatosis defined by score ≥2 (S≥2) and moderate/severe steatosis by score ≥4 (S≥4). Performance of CAP and FLI for diagnosing steatosis compared with US was assessed using areas under receiver operating characteristic curves (AUROC). RESULTS: From 219 consecutive individuals studied, 13 (5.9%) excluded because of failure/unreliable liver stiffness measurements. Steatosis prevalence: S≥2 38.4% and S≥4 12.1%. CAP significantly correlated with steatosis (ρ = 0.73, P < 0.0001), steatosis score (ρ = 0.76; P < 0.0001), FLI (ρ = 0.69), waist circumference (ρ = 0.62), body mass index (ρ = 0.55), triglyceride (ρ = 0.49), HOMA-IR (ρ = 0.26), alcohol consumption (ρ = 0.24) and cholesterol (ρ = 0.19), not with liver stiffness measurements. Using CAP and FLI, AUROC's were 0.94 (95% CI 0.91-0.97, P < 0.001) and 0.91 for S≥2; 0.95 (95% CI 0.90-0.99, P < 0.001) and 0.93 for S≥4 respectively. Optimal cut-off value of CAP and FLI were 243 dB/m and 48 for S≥2; 303.5 dB/m and 62 for S≥4 respectively. CONCLUSION: Controlled attenuation parameter and FLI seem promising tools for screening and steatosis quantification in the general population. Larger studies are needed for validation.