A SAM-key domain required for enzymatic activity of the Fun30 nucleosome remodeler.

Fun30 is the prototype of the Fun30-SMARCAD1-ETL subfamily of nucleosome remodelers involved in DNA repair and gene silencing. These proteins appear to act as single-subunit nucleosome remodelers, but their molecular mechanisms are, at this point, poorly understood. Using multiple sequence alignment and structure prediction, we identify an evolutionarily conserved domain that is modeled to contain a SAM-like fold with one long, protruding helix, which we term SAM-key. Deletion of the SAM-key within budding yeast Fun30 leads to a defect in DNA repair and gene silencing similar to that of the fun30Δ mutant. In vitro, Fun30 protein lacking the SAM-key is able to bind nucleosomes but is deficient in DNA-stimulated ATPase activity and nucleosome sliding and eviction. A structural model based on AlphaFold2 prediction and verified by crosslinking-MS indicates an interaction of the long SAM-key helix with protrusion I, a subdomain located between the two ATPase lobes that is critical for control of enzymatic activity. Mutation of the interaction interface phenocopies the domain deletion with a lack of DNA-stimulated ATPase activation and a nucleosome-remodeling defect, thereby confirming a role of the SAM-key helix in regulating ATPase activity. Our data thereby demonstrate a central role of the SAM-key domain in mediating the activation of Fun30 catalytic activity, thus highlighting the importance of allosteric activation for this class of enzymes.

Probiotic, technological, and health-related characteristics of lactobacilli isolated from breast milk.

AIMS: Isolation and characterization of lactobacilli from human milk and determination of their probiotic, technological, and in vitro health-promoting properties with a view to their potential use in food fermentation. METHODS AND RESULTS: Seven lactobacilli isolates were obtained from human milk and identified as Lacticaseibacillus paracasei (isolates BM1-BM6) and Lactobacillus gasseri (BM7). The isolates were examined in vitro for their technological, probiotic, and health-promoting potential. Overall, all isolates showed important technological properties based on the ability to grow in milk whey, a high to moderate acidification capacity and the absence of undesirable enzymatic activities. Lacticaseibacillus gasseri (BM7) differed from the L. paracasei isolates by the absence of several glycosidases and the inability to ferment lactose. Isolates L. paracasei BM3 and BM5 produced exopolysaccharides (EPS) from lactose. All isolates showed probiotic potential as they were tolerant to simulated gastrointestinal conditions, had high cell surface hydrophobicity, had not acquired resistance to relevant antibiotics and had no virulence characteristics. All L. paracasei showed high antimicrobial activity against various pathogenic bacteria and fungi, while L. gasseri showed a narrower spectrum of antimicrobial activity. All isolates showed health-promoting potential in vitro, as evidenced by high cholesterol-lowering activity, high ACE inhibitory activity and marked antioxidant activity. CONCLUSIONS: All strains showed excellent probiotic and technological properties for use in lactic ferments.

Mob4 is essential for spermatogenesis in Drosophila melanogaster.

Gamete formation is essential for sexual reproduction in metazoans. Meiosis in males gives rise to spermatids that must differentiate and individualize into mature sperm. In Drosophila melanogaster, individualization of interconnected spermatids requires the formation of individualization complexes that synchronously move along the sperm bundles. Here, we show that Mob4, a member of the Mps-one binder family, is essential for male fertility but has no detectable role in female fertility. We show that Mob4 is required for proper axonemal structure and its loss leads to male sterility associated with defective spermatid individualization and absence of mature sperm in the seminal vesicles. Transmission electron micrographs of developing spermatids following mob4RNAi revealed expansion of the outer axonemal microtubules such that the 9 doublets no longer remained linked to each other and defective mitochondrial organization. Mob4 is a STRIPAK component, and male fertility is similarly impaired upon depletion of the STRIPAK components, Strip and Cka. Expression of the human Mob4 gene rescues all phenotypes of Drosophila mob4 downregulation, indicating that the gene is evolutionarily and functionally conserved. Together, this suggests that Mob4 contributes to the regulation of the microtubule- and actin-cytoskeleton during spermatogenesis through the conserved STRIPAK complex. Our study advances the understanding of male infertility by uncovering the requirement for Mob4 in sperm individualization.

RNA-DNA hybrids prevent resection at dysfunctional telomeres.

At critically short telomeres, stabilized TERRA RNA-DNA hybrids drive homology-directed repair (HDR) to delay replicative senescence. However, even at long- and intermediate-length telomeres, not subject to HDR, transient TERRA RNA-DNA hybrids form, suggestive of additional roles. We report that telomeric RNA-DNA hybrids prevent Exo1-mediated resection when telomeres become non-functional. We used the well-characterized cdc13-1 allele, where telomere resection can be induced in a temperature-dependent manner, to demonstrate that ssDNA generation at telomeres is either prevented or augmented when RNA-DNA hybrids are stabilized or destabilized, respectively. The viability of cdc13-1 cells is affected by the presence or absence of hybrids accordingly. Telomeric hybrids do not affect the shortening rate of bulk telomeres. We suggest that TERRA hybrids require dynamic regulation to drive HDR at short telomeres; hybrid presence may initiate HDR through replication stress, whereby their removal allows strand resection.

Thioredoxin Reductase Inhibitors as Potential Antitumors: Mercury Compounds Efficacy in Glioma Cells.

Glioblastoma multiforme (GBM) is the most aggressive and common form of glioma. GBM, like many other tumors, expresses high levels of redox proteins, such as thioredoxin (Trx) and thioredoxin reductase (TrxR), allowing tumor cells to cope with high levels of reactive oxygen species (ROS) and resist chemotherapy and radiotherapy. Thus, tackling the activity of these enzymes is a strategy to reduce cell viability and proliferation and most importantly achieve tumor cell death. Mercury (Hg) compounds are among the most effective inhibitors of TrxR and Trx due to their high affinity for binding thiols and selenols. Moreover, organomercurials such as thimerosal, have a history of clinical use in humans. Thimerosal effectively crosses the blood-brain barrier (BBB), thus reaching effective concentrations for the treatment of GBM. Therefore, this study evaluated the effects of thimerosal (TmHg) and its metabolite ethylmercury (EtHg) over the mouse glioma cell line (GL261), namely, the inhibition of the thioredoxin system and the occurrence of oxidative cellular stress. The results showed that both TmHg and EtHg increased oxidative events and triggered cell death primarily by apoptosis, leading to a significant reduction in GL261 cell viability. Moreover, the cytotoxicity of TmHg and ETHg in GL261 was significantly higher when compared to temozolomide (TMZ). These results indicate that EtHg and TmHg have the potential to be used in GBM therapy since they strongly reduce the redox capability of tumor cells at exceedingly low exposure levels.

Suplementação de Vitamina D Induz Remodelação Cardíaca em Ratos: Associação com a Proteína de Interação com a Tiorredoxina e a Tiorredoxina / Vitamin D Supplementation Induces Cardiac Remodeling in Rats: Association with Thioredoxin-Interacting Protein and Thioredoxin

Resumo Fundamento: A vitamina D (VD) tem um importante papel na função cardíaca. No entanto, a vitamina exerce uma curva "dose-resposta" bifásica na fisiopatologia cardiovascular e pode causar efeitos deletérios, mesmo em doses não tóxicas. A VD exerce suas funções celulares ligando-se ao seu receptor. Ainda, a expressão da proteína de interação com a tiorredoxina (TXNIP) é positivamente regulada pela VD. A TXNIP modula diferentes visa de sinalização celular que podem ser importantes para a remodelação cardíaca. Objetivos: Avaliar se a suplementação com VD leva à remodelação cardíaca, e se a TXNIP e a tiorredoxina (Trx) estão associadas com esse processo. Métodos: Duzentos e cinquenta ratos Wistar machos foram alocados em três grupos: controle (C, n=21), sem suplementação com VD; VD3 (n = 22) e VD10 (n=21), suplementados com 3,000 e 10,000 UI de VD/ kg de ração, respectivamente, por dois meses. Os grupos foram comparados por análise de variância (ANOVA) com um fator e teste post hoc de Holm-Sidak (variáveis com distribuição normal), ou pelo teste de Kruskal-Wallis e análise post-hoc de Dunn. O nível de significância para todos os testes foi de 5%. Resultados: A expressão de TXNIP foi mais alta e a atividade do Trx foi mais baixa no grupo VD10. Os animais que receberam suplementação com VD apresentaram aumento de hidroperóxido lipídico e diminuição de superóxido dismutase e glutationa peroxidase. A proteína Bcl-2 foi mais baixa no grupo VD10. Observou-se uma diminuição na β-oxidação de ácidos graxos, no ciclo do ácido tricarboxílico, na cadeia transportadora de elétrons, e um aumento na via glicolítica. Conclusão: A suplementação com VD levou à remodelação cardíaca e esse processo pode ser modulado por TXNIP e Trx, e consequentemente por estresse oxidativo.

Abstract Background: Vitamin D (VD) has been shown to play an important role in cardiac function. However, this vitamin exerts a biphasic "dose response" curve in cardiovascular pathophysiology and may cause deleterious effects, even in non-toxic doses. VD exerts its cellular functions by binding to VD receptor. Additionally, it was identified that the thioredoxin-interacting protein (TXNIP) expression is positively regulated by VD. TXNIP modulate different cell signaling pathways that may be important for cardiac remodeling. Objective: To evaluate whether VD supplementation lead to cardiac remodeling and if TXNIP and thioredoxin (Trx) proteins are associated with the process. Methods: A total of 250 Male Wistar rats were allocated into three groups: control (C, n=21), with no VD supplementation; VD3 (n = 22) and VD10 (n=21), supplemented with 3,000 and 10,000 IU of VD/ kg of chow respectively, for two months. The groups were compared by one-way analysis of variance (ANOVA) and Holm-Sidak post hoc analysis, (variables with normal distribution), or by Kruskal-Wallis test and Dunn's test post hoc analysis. The significance level for all tests was 5%. Results: TXNIP protein expression was higher and Trx activity was lower in VD10. The animals supplemented with VD showed increased lipid hydroperoxide and decreased superoxide dismutase and glutathione peroxidase. The protein Bcl-2 was lower in VD10. There was a decrease in fatty acid β-oxidation, tricarboxylic acid cycle and electron transport chain with shift to increase in glycolytic pathway. Conclusion: VD supplementation led to cardiac remodeling and this process may be modulated by TXNIP and Trx proteins and consequently oxidative stress.

Feeding practices and early weaning in the neonatal period: a cohort study.

OBJECTIVE: To describe feeding practices and the risk factors for the mixed breastfeeding and early weaning in the neonatal period. METHODS: Cohort study, which we collected socioeconomic, demographic, health care and feeding data from 415 mother/child binomials born in four public maternity hospitals in Natal/Brazil. They were followed-up at 48 hours, 7 and 28 days after birth. The association was established using Pearson's Chi-square test and Poisson's regression, after adjusting it to other variables. RESULTS: The prevalence of mixed breastfeeding in the first 2 days was 47,2% and early weaning in 7 and 28 days was 8,4% and 16,2% in that order. The main reasons for mixed breastfeeding and early weaning were: colostrum deficiency (33.8%), difficulty in latching/sucking (23.5%) and "little milk" (70.0%). The use of formula/milk/porridge remained associated with maternal age ≤ 20 years (RR = 0.64; 95%CI: 0.47-0.86), age 20-29 years (RR = 0,70; 95%CI: 0,57-0,87), primiparity (RR = 1.37; 95%CI: 1.11-1.60) and cesarean delivery (RR = 1.20; 95%CI: 1.00-1.45) at 2 days; absence of paternal support (RR = 4.98; 95%CI: 2.54-9.79) and pacifier use (RR = 3.21; 95%CI: 1.63-6.32) at 7 days; and only pacifier use (RR = 2.48; 95%CI: 1.53-4.02) at 28 days. CONCLUSIONS: Early weaning was associated with maternal and health care factors, thus suggesting the need to readjust good practices and educational actions to achieve the exclusive offer to the maternal breast in the neonatal period.

Prevalence of the five newborn screening tests.

Neonatal screening is essential for child health and has the following purposes: (1) pulse oximetry screening to evaluate congenital heart diseases; (2) red reflex examination to investigate eye diseases; (3) newborn hearing screening test to evaluate congenital hearing diseases; (4) tongue test to evaluate the lingual frenulum and identify communication and feeding problems; (5) the Guthrie test to screen for metabolic diseases. This study investigated the prevalence of the five neonatal screening tests and its associated institutional and socio-cultural factors using a cross-sectional study with 415 mother and baby binomials from public maternity hospitals in Natal, RN, Brazil in 2019. Pearson's chi-squared, Mann-Whitney and Poisson regression tests were used, with a significance of p ≤ 0.05 and a 95% confidence interval. The sample loss was 71 mothers (17%). The prevalence in the first week and at the end of 28 days was 93% and 99.5% (pulse oximetry screening), 60% and 97.6% (red reflex examination), 71.9% and 93.6% (Guthrie test), 35.5% and 68.2% (hearing screening test), and 19% and 48.9% (tongue test). Only 152 newborns (36.6%) underwent all five tests. The performance of the tests was associated in the final model (p ≤ 0.05) with the residence of the mothers in the state capital (PR = 1.36; 95% CI = 1.18-1.56) and the provision of guidance for mothers about the five tests in maternity hospitals (PR = 1.30; 95% CI = 1.08-1.67). None of the tests met full coverage, and regional inequities were identified indicating the need to restructure the institutions, training and qualification procedures to improve of the work processes and longitudinal care.

Prevalence on the complement in offering food to newborns / Prevalência da oferta de complemento alimentar para o recém-nascido

Abstract Objectives: to identify the prevalence and determining factors of the complements in offering food to newborns. Methods: across-sectional study nested to a cohort study that assessed newborn care in four public maternity hospitals in Natal/Brazil. Sample was composed by 415 mothers and full-term newborns, with appropriate weight for gestational age and Apgar scores in 1st and 5th minutes ≥ 7. In order to analyze the determining factors, we used Poisson's regression. Results: from 415 newborns, 51.3% received feeding complements (57.6% in the first hour of life), of which 92% was infant formula. Only 50.7% of those complement in offering food were prescribed by physicians. Colostrum deficiency was the main reason to be indicated (33.8%). Maternal age ≤ 20 years old (PR=0.64; CI95%=0.47-0.86) and between 2030 years old (PR=0.70; CI95%=0.57-0.87)comparing to women older than 30 were shown as protective factors, while being primiparous (PR=1.37; CI95%=1.11-1.60) and had cesarean section (PR=1.2; CI95%=1.00-1.45) as risk factors. Conclusion: maternal characteristics are associated with the complement in offering food to the newborn in the first hours of life. The high prevalence shows the need for interventions that minimize the inadequate offer of infant formula and promote exclusive breastfeeding before hospital discharge.

Resumo Objetivos: identificar a prevalência e os fatores determinantes da oferta do complemento alimentar para o recém-nascido. Métodos: estudo transversal, aninhado a um estudo de coorte que avaliou a assistência ao neonato em quatro maternidades públicas de Natal/Brasil. Amostra composta de 415 mães, e recém-nascidos à termo, com peso adequado para idade gestacional e Apgar no 1º e 5º minuto ≥ 7. Para analisar os fatores determinantes, foi utilizado a regressão de Poisson. Resultados: dos 415 recém-nascidos, 51,3% receberam complemento (57,6% na primeira hora de vida), dos quais 92% com fórmula infantil. Destes, apenas 50,7% foi prescrito pelo médico. A deficiência de colostro foi o principal motivo de indicação (33,8%). A idade materna ≤ 20 anos (RP=0,64; IC95%=0,47-0,86) e entre 20-30 anos (RP=0,70; IC95%=0,57-0,87, em comparação com mulheres acima de 30 anos, mostrou-se como fator de proteção, enquanto ser primípara (RP=1,37; IC95%=1,11-1,60) e o parto cesárea (RP=1,2; IC95%=1,00-1,45) como fatores de risco. Conclusão: as características maternas e assistenciais estão associadas à oferta de complemento alimentar ao recém-nascido nas primeiras horas de vida. A alta prevalência mostra a necessidade de intervenções que minimizem a oferta inadequada de fórmula infantil, e promovam o aleitamento materno exclusivo antes da alta hospitalar.

Vitamin D Supplementation Induces Cardiac Remodeling in Rats: Association with Thioredoxin-Interacting Protein and Thioredoxin. / Suplementação de Vitamina D Induz Remodelação Cardíaca em Ratos: Associação com a Proteína de Interação com a Tiorredoxina e a Tiorredoxina.

BACKGROUND: Vitamin D (VD) has been shown to play an important role in cardiac function. However, this vitamin exerts a biphasic "dose response" curve in cardiovascular pathophysiology and may cause deleterious effects, even in non-toxic doses. VD exerts its cellular functions by binding to VD receptor. Additionally, it was identified that the thioredoxin-interacting protein (TXNIP) expression is positively regulated by VD. TXNIP modulate different cell signaling pathways that may be important for cardiac remodeling. OBJECTIVE: To evaluate whether VD supplementation lead to cardiac remodeling and if TXNIP and thioredoxin (Trx) proteins are associated with the process. METHODS: A total of 250 Male Wistar rats were allocated into three groups: control (C, n=21), with no VD supplementation; VD3 (n = 22) and VD10 (n=21), supplemented with 3,000 and 10,000 IU of VD/ kg of chow respectively, for two months. The groups were compared by one-way analysis of variance (ANOVA) and Holm-Sidak post hoc analysis, (variables with normal distribution), or by Kruskal-Wallis test and Dunn's test post hoc analysis. The significance level for all tests was 5%. RESULTS: TXNIP protein expression was higher and Trx activity was lower in VD10. The animals supplemented with VD showed increased lipid hydroperoxide and decreased superoxide dismutase and glutathione peroxidase. The protein Bcl-2 was lower in VD10. There was a decrease in fatty acid ß-oxidation, tricarboxylic acid cycle and electron transport chain with shift to increase in glycolytic pathway. CONCLUSION: VD supplementation led to cardiac remodeling and this process may be modulated by TXNIP and Trx proteins and consequently oxidative stress.

FUNDAMENTO: A vitamina D (VD) tem um importante papel na função cardíaca. No entanto, a vitamina exerce uma curva "dose-resposta" bifásica na fisiopatologia cardiovascular e pode causar efeitos deletérios, mesmo em doses não tóxicas. A VD exerce suas funções celulares ligando-se ao seu receptor. Ainda, a expressão da proteína de interação com a tiorredoxina (TXNIP) é positivamente regulada pela VD. A TXNIP modula diferentes visa de sinalização celular que podem ser importantes para a remodelação cardíaca. OBJETIVOS: Avaliar se a suplementação com VD leva à remodelação cardíaca, e se a TXNIP e a tiorredoxina (Trx) estão associadas com esse processo. MÉTODOS: Duzentos e cinquenta ratos Wistar machos foram alocados em três grupos: controle (C, n=21), sem suplementação com VD; VD3 (n = 22) e VD10 (n=21), suplementados com 3,000 e 10,000 UI de VD/ kg de ração, respectivamente, por dois meses. Os grupos foram comparados por análise de variância (ANOVA) com um fator e teste post hoc de Holm-Sidak (variáveis com distribuição normal), ou pelo teste de Kruskal-Wallis e análise post-hoc de Dunn. O nível de significância para todos os testes foi de 5%. RESULTADOS: A expressão de TXNIP foi mais alta e a atividade do Trx foi mais baixa no grupo VD10. Os animais que receberam suplementação com VD apresentaram aumento de hidroperóxido lipídico e diminuição de superóxido dismutase e glutationa peroxidase. A proteína Bcl-2 foi mais baixa no grupo VD10. Observou-se uma diminuição na ß-oxidação de ácidos graxos, no ciclo do ácido tricarboxílico, na cadeia transportadora de elétrons, e um aumento na via glicolítica. CONCLUSÃO: A suplementação com VD levou à remodelação cardíaca e esse processo pode ser modulado por TXNIP e Trx, e consequentemente por estresse oxidativo.

Hypoglycemic episodes in hospitalized people with diabetes in Portugal: the HIPOS-WARD study.

BACKGROUND: We intended to estimate the proportion hypoglycemic/hyperglycemic emergency episodes in treated diabetes mellitus (DM) patients admitted to a hospital ward, and calculate the prevalence of risk factors for hypoglycemia and diabetic complications. METHODS: In this cross-sectional, multicentered study, the observational data was collected by physicians from patient's hospitalization to discharge/death. Statistical tests were 2-tailed considering 5% significance level. RESULTS: There were 646 ward admissions due to hyperglycemic emergencies and 176 hypoglycemic episodes with a ratio hypoglycemia/hyperglycemia 0.27 for all DM patients. In T2DM patients the ratio was 0.38. These were mainly female (55.1%), functionally dependent (61.4%) and retired/disabled (73.1%). Median age was 75 years and median duration of disease 11 years. Half the patients were on insulin-based therapy and 30.1% on secretagogue-based therapy. Approximately 57% of patients needed occasional/full assistance to manage the disease. The most frequent risk factor for hypoglycemia was polypharmacy (85.0%). Hypoglycemia in the 12 months before admission was higher in insulin-based therapy patients (66.1%; p = 0.001). CONCLUSIONS: Hyperglycemic emergencies are the most frequent cause of hospitalization in Portugal, although severe hypoglycemic events represent a health and social problem in elderly/frail patients. There is still the need to optimize therapy in terms of the potential for hypoglycemia in this patient group and a review of anti-hyperglycemic agents to add on to insulin.

The role of glucose metabolism and insulin resistance in cardiac remodelling induced by cigarette smoke exposure.

The aim of this study is to evaluate whether the alterations in glucose metabolism and insulin resistance are mechanisms presented in cardiac remodelling induced by the toxicity of cigarette smoke. Male Wistar rats were assigned to the control group (C; n = 12) and the cigarette smoke-exposed group (exposed to cigarette smoke over 2 months) (CS; n = 12). Transthoracic echocardiography, blood pressure assessment, serum biochemical analyses for catecholamines and cotinine, energy metabolism enzymes activities assay; HOMA index (homeostatic model assessment); immunohistochemistry; and Western blot for proteins involved in energy metabolism were performed. The CS group presented concentric hypertrophy, systolic and diastolic dysfunction, and higher oxidative stress. It was observed changes in energy metabolism, characterized by a higher HOMA index, lower concentration of GLUT4 (glucose transporter 4) and lower 3-hydroxyl-CoA dehydrogenase activity, suggesting the presence of insulin resistance. Yet, the cardiac glycogen was depleted, phosphofructokinase (PFK) and lactate dehydrogenase (LDH) increased, with normal pyruvate dehydrogenase (PDH) activity. The activity of citrate synthase, mitochondrial complexes and ATP synthase (adenosine triphosphate synthase) decreased and the expression of Sirtuin 1 (SIRT1) increased. In conclusion, exposure to cigarette smoke induces cardiac remodelling and dysfunction. The mitochondrial dysfunction and heart damage induced by cigarette smoke exposure are associated with insulin resistance and glucose metabolism changes.

Feeding practices and early weaning in the neonatal period: a cohort study

ABSTRACT OBJECTIVE To describe feeding practices and the risk factors for the mixed breastfeeding and early weaning in the neonatal period. METHODS Cohort study, which we collected socioeconomic, demographic, health care and feeding data from 415 mother/child binomials born in four public maternity hospitals in Natal/Brazil. They were followed-up at 48 hours, 7 and 28 days after birth. The association was established using Pearson's Chi-square test and Poisson's regression, after adjusting it to other variables. RESULTS The prevalence of mixed breastfeeding in the first 2 days was 47,2% and early weaning in 7 and 28 days was 8,4% and 16,2% in that order. The main reasons for mixed breastfeeding and early weaning were: colostrum deficiency (33.8%), difficulty in latching/sucking (23.5%) and "little milk" (70.0%). The use of formula/milk/porridge remained associated with maternal age ≤ 20 years (RR = 0.64; 95%CI: 0.47-0.86), age 20-29 years (RR = 0,70; 95%CI: 0,57-0,87), primiparity (RR = 1.37; 95%CI: 1.11-1.60) and cesarean delivery (RR = 1.20; 95%CI: 1.00-1.45) at 2 days; absence of paternal support (RR = 4.98; 95%CI: 2.54-9.79) and pacifier use (RR = 3.21; 95%CI: 1.63-6.32) at 7 days; and only pacifier use (RR = 2.48; 95%CI: 1.53-4.02) at 28 days. CONCLUSIONS Early weaning was associated with maternal and health care factors, thus suggesting the need to readjust good practices and educational actions to achieve the exclusive offer to the maternal breast in the neonatal period.

Hospitalization Costs Due to Hypoglycemia in Patients with Diabetes: A Microcosting Approach.

INTRODUCTION: Hypoglycemia leading to hospitalization is associated with adverse economic outcomes, although the real burden is unknown. The HIPOS-WARD (Hypoglycemia In Portugal Observational Study-Ward) aimed to characterize ward admissions due to hypoglycemia episodes in treated patients with diabetes and assess their economic impact to the National Health System. METHODS: Observational, cross-sectional study, conducted in 16 Portuguese centers for 22 months. The applied microcosting approach was based on healthcare resource data, collected from patients' charts upon ward admission until discharge, and unitary costs from official/public data sources. Absenteeism was also estimated for active workers on the basis of the human capital approach. RESULTS: Of the 176 patients with diabetes mellitus enrolled, 86% had type 2 diabetes. Half of the patients (50.0%) were on insulin-based therapy, followed by 30.1% on a secretagogue-based regimen, 9.7% on non-secretagogue therapy, and 10.2% on a combination of insulin and secretagogue. Overall mean costs per patient were medication, 45.45 €; laboratory analysis, 218.14 €; examinations, 64.91 €; physician and nurse time, 268.55 € and 673.39 €, respectively. Bed occupancy was the main cost driver (772.09 €) and indirect cost averaged 140.44 €. Overall, the cost per hypoglycemia episode leading to hospitalization averaged 2042.52 € (range 194.76-16,762.87 €). Patients treated with insulin-based regimens (2267.76 €) and type 2 diabetes (2051.29 €) had the highest mean costs. The mean cost increased with repeated hypoglycemic events (2191.67 €), correlated complications (2109.26 €), and death (5253.38 €). CONCLUSION: HIPOS-WARD's findings confirm and support both the substantial clinical and economic impact of hospitalization due to hypoglycemia in Portugal.

Benefits of the consumption of Brazil nut (Bertholletia excelsa) extract in male reproductive parameters of streptozotocin-induced diabetic rats.

OBJECTIVE: The objective of this study was to evaluate the effects of intake of Brazil nut extract (BN) or sodium selenite solution on reproductive parameters of male diabetic animals. METHODS: A total of 48 Wistar rats were distributed into six groups: diabetes (n = 8); diabetes and Brazil nut extract (n = 8); diabetes and sodium selenite (Na2SeO3) (n = 8); Brazil nut extract (n = 8); sodium selenite (n = 8) and control (n = 8). A single dose of streptozotocin (65 mg/kg) was injected intravenously to the rats to induce diabetes. BN or Na2SeO3 were administered by gavage for 56 days. RESULTS: The diabetes caused critical alterations on body mass gain, reproductive parameters and antioxidant capacity. Treatments with both BN or Na2SeO3 were able to increase significantly the glutathione peroxidase and the daily sperm production, both in diabetic (p < 0.01 and p < 0.05) and in healthy animals (p < 0.01 and p < 0.05). CONCLUSION: The Brazil nut extract and sodium selenite were able to improve some reproductive parameters of diabetic rats. Moreover, we could infer that this effect is probably due to the natural selenium content of the BN.

RNase H1 and H2 Are Differentially Regulated to Process RNA-DNA Hybrids.

RNA-DNA hybrids are tightly regulated to ensure genome integrity. The RNase H enzymes RNase H1 and H2 contribute to chromosomal stability through the removal of RNA-DNA hybrids. Loss of RNase H2 function is implicated in human diseases of the nervous system and cancer. To better understand RNA-DNA hybrid dynamics, we focused on elucidating the regulation of the RNase H enzymes themselves. Using yeast as a model system, we demonstrate that RNase H1 and H2 are controlled in different manners. RNase H2 has strict cell cycle requirements, in that it has an essential function in G2/M for both R-loop processing and ribonucleotide excision repair. RNase H1, however, can function independently of the cell cycle to remove R-loops and appears to become activated in response to high R-loop loads. These results provide us with a more complete understanding of how and when RNA-DNA hybrids are acted upon by the RNase H enzymes.

The role of Xist-mediated Polycomb recruitment in the initiation of X-chromosome inactivation.

Xist RNA has been established as the master regulator of X-chromosome inactivation (XCI) in female eutherian mammals, but its mechanism of action remains unclear. By creating novel Xist-inducible mutants at the endogenous locus in male mouse embryonic stem (ES) cells, we dissect the role of the conserved A-B-C-F repeats in the initiation of XCI. We find that transcriptional silencing can be largely uncoupled from Polycomb repressive complex 1 and complex 2 (PRC1/2) recruitment, which requires B and C repeats. Xist ΔB+C RNA specifically loses interaction with PCGF3/5 subunits of PRC1, while binding of other Xist partners is largely unaffected. However, a slight relaxation of transcriptional silencing in Xist ΔB+C indicates a role for PRC1/2 proteins in early stabilization of gene repression. Distinct modules within the Xist RNA are therefore involved in the convergence of independent chromatin modification and gene repression pathways. In this context, Polycomb recruitment seems to be of moderate relevance in the initiation of silencing.

Unveiling the Insular Lobe of Reil: Neurophysiological and Anatomical Features

The insular lobe has long been investigated, from its anatomical descriptions to its neurophysiological activity. Located in a central location, the insular lobe participates in several afferent and efferent pathways, forming part of the eloquent and fundamental structures that make up the central core of the brain. The lobe of the insula has participation in language function, such as speech, sensory (e.g., taste), limbic, autonomic (visceral), also forming part of complex associative circuits, including part of the circuits of mirror neurons. Several functional descriptions attributed to the insular lobe have beenmade in patients suffering fromcerebrovascular diseases, as well as in those with epilepsy. Much progress and many descriptions have also been made in patients with tumors. Despite much information already available about the insular lobe, it is likely that much will be discovered in the coming years.

Lipid-lowering and antiatherogenic effects of Vitex megapotamica (Spreng.) Moldenke in a mice experimental model.

ETHNOPHARMACOLOGICAL RELEVANCE: Vitex megapotamica (Spreng.) Moldenke is a deciduous tree, native of South America. Its leaves are traditionally used to treat cardiovascular diseases. This activity is related to the presence of flavonoids, the major compounds of the crude extract. AIM OF THE STUDY: This study investigated the effects of the oral administration of crude extract and standardized fractions from V. megapotamica leaves on lipid profile and on the formation of atherosclerotic plaque in C57BL/6 LDLr-KO mice treated with high-fat diet (HFD). MATERIALS AND METHODS: Male C57BL/6 LDLr-KO mice were fed with HFD (cholesterol, 1.25%) for 30 days. They were treated with hydroethanolic extract (500 or 1000mg/kg/day) or fractions (125 or 250mg/kg/day). After 30 days of treatment, it was evaluated the serum lipid profile, atherogenic index, and atherosclerotic plaque. RESULTS: All doses of the hydroethanolic extract reduced significantly the levels of total cholesterol, triglycerides, LDL-c and the atherogenic index. The n-butanolic fraction also reduced significantly the levels of total cholesterol, triglycerides, LDL-c and the atherogenic index, at all doses, with exception for the triglycerides, which only the lower dose was effective. The residual fraction reduced significantly the levels of total cholesterol, LDL-c and the atherogenic index, at all doses, with exception for the atherogenic index, which only the higher dose was effective. The atherosclerotic plaque formation was impaired only by the lower dose of the hydroethanolic extract. CONCLUSIONS: Overall, our data suggest that V. megapotamica has potential for the treatment of dyslipidemias.

Short (16-mer) locked nucleic acid splice-switching oligonucleotides restore dystrophin production in Duchenne Muscular Dystrophy myotubes.

Splice-switching antisense oligonucleotides (SSOs) offer great potential for RNA-targeting therapies, and two SSO drugs have been recently approved for treating Duchenne Muscular Dystrophy (DMD) and Spinal Muscular Atrophy (SMA). Despite promising results, new developments are still needed for more efficient chemistries and delivery systems. Locked nucleic acid (LNA) is a chemically modified nucleic acid that presents several attractive properties, such as high melting temperature when bound to RNA, potent biological activity, high stability and low toxicity in vivo. Here, we designed a series of LNA-based SSOs complementary to two sequences of the human dystrophin exon 51 that are most evolutionary conserved and evaluated their ability to induce exon skipping upon transfection into myoblasts derived from a DMD patient. We show that 16-mers with 60% of LNA modification efficiently induce exon skipping and restore synthesis of a truncated dystrophin isoform that localizes to the plasma membrane of patient-derived myotubes differentiated in culture. In sum, this study underscores the value of short LNA-modified SSOs for therapeutic applications.