Regulation of Myc transcription by an enhancer cluster dedicated to pluripotency and early embryonic expression.

MYC plays various roles in pluripotent stem cells, including the promotion of somatic cell reprogramming to pluripotency, the regulation of cell competition and the control of embryonic diapause. However, how Myc expression is regulated in this context remains unknown. The Myc gene lies within a ~ 3-megabase gene desert with multiple cis-regulatory elements. Here we use genomic rearrangements, transgenesis and targeted mutation to analyse Myc regulation in early mouse embryos and pluripotent stem cells. We identify a topologically-associated region that homes enhancers dedicated to Myc transcriptional regulation in stem cells of the pre-implantation and early post-implantation embryo. Within this region, we identify elements exclusively dedicated to Myc regulation in pluripotent cells, with distinct enhancers that sequentially activate during naive and formative pluripotency. Deletion of pluripotency-specific enhancers dampens embryonic stem cell competitive ability. These results identify a topologically defined enhancer cluster dedicated to early embryonic expression and uncover a modular mechanism for the regulation of Myc expression in different states of pluripotency.

An adapted cardioprotective diet with or without phytosterol and/or krill oil supplement in familial hypercholesterolemia: A pilot study protocol

BACKGROUND AND AIMS: A healthy diet is one of the pillars of familial hypercholesterolemia (FH) treatment. However, the best dietary pattern and indication for specific supplementation have not been established. Our aim is to conduct a pilot study to assess the effect of an adapted cardioprotective diet with or without phytosterol and/or krill oil supplement in participants with a probable or definitive diagnosis of FH, treated with moderate/high potency statins. METHODS: A national, multicenter, factorial, and parallel placebocontrolled randomized clinical trial with a superiority design and 1:1:1:1 allocation rate will be conducted. The participants will undergo whole exome sequencing and be allocated into four treatment groups: 1) a cardioprotective diet adapted for FH (DICAFH) þ phytosterol placebo þ krill oil placebo; 2) DICA-FH þ phytosterol 2 g/day þ krill oil placebo; 3) DICA-FH þ phytosterol placebo þ krill oil 2 g/day; or 4) DICA-FH þ phytosterol 2 g/day þ krill oil 2 g/day. The primary outcomes will be low-density lipoprotein (LDL)-cholesterol and lipoprotein (a) levels and adherence to treatment after a 120-day follow-up. LDL- and high-density lipoprotein (HDL)-cholesterol subclasses, untargeted lipidomics analysis, adverse events, and protocol implementation components will also be assessed. RESULTS: A total of 58 participants were enrolled between May e August 2023. After the end of the follow-up period, the efficacy and feasibility results of this pilot study will form the basis of the design of a large-scale randomized clinical trial. CONCLUSIONS: This study's overall goal is to recommend dietary treatment strategies in the context of FH.

P53 and BCL-2 family proteins PUMA and NOXA define competitive fitness in pluripotent cell competition.

Cell Competition is a process by which neighboring cells compare their fitness. As a result, viable but suboptimal cells are selectively eliminated in the presence of fitter cells. In the early mammalian embryo, epiblast pluripotent cells undergo extensive Cell Competition, which prevents suboptimal cells from contributing to the newly forming organism. While competitive ability is regulated by MYC in the epiblast, the mechanisms that contribute to competitive fitness in this context are largely unknown. Here, we report that P53 and its pro-apoptotic targets PUMA and NOXA regulate apoptosis susceptibility and competitive fitness in pluripotent cells. PUMA is widely expressed specifically in pluripotent cells in vitro and in vivo. We found that P53 regulates MYC levels in pluripotent cells, which connects these two Cell Competition pathways, however, MYC and PUMA/NOXA levels are independently regulated by P53. We propose a model that integrates a bifurcated P53 pathway regulating both MYC and PUMA/NOXA levels and determines competitive fitness.

Effects of Whey Protein Isolate on Body Composition, Muscle Mass, and Strength of Chronic Heart Failure Patients: A Randomized Clinical Trial.

Heart failure (HF) is associated with a reduction of skeletal muscle mass. Whey protein isolate (WPI) has been beneficial in increasing muscle mass and strength, in addition to improving body composition. The goal of this research was to evaluate the effect of WPI on the body composition, muscle mass, and strength of chronic HF patients. For this purpose, twenty-five patients of both genders with predominantly NYHA I functional class and a median age of 65.5 (60.5-71.0) years were used to conduct a randomized, single-blind, placebo-controlled clinical trial and received 30 g per day of WPI for 12 weeks. Anthropometric measurements, body composition analysis, and biochemical exams were performed at the beginning and end of the study. An increase in skeletal muscle mass was observed in the intervention group after 12 weeks. A reduction in waist circumference, body fat percentage, and an increase in skeletal muscle index was observed when compared to the placebo group. No significant effect on muscle strength was observed after 12 weeks of intervention. These data demonstrate that WPI consumption contributed to the increase of skeletal muscle mass, strength, and reduction of body fat in HF patients.

First Description of Acaricide Resistance in Populations of Rhipicephalus microplus Tick from the Lower Amazon, Brazil.

There is limited information on the resistance to acaricidal drugs of the cattle tick Rhipicephalus microplus in the Lower Amazon region. Thus, we aimed to determine the efficiency of three widely used acaricide products (Amitraz, Cypermethrin, and Deltamethrin) in the control of this tick species. The adult immersion test was used on engorged female ticks sampled on farms in the Lower Amazon region, Brazil. For the test, homogeneous batches of 10 engorged females were placed in Petri dishes and immersed in the tested acaricidal drugs, using four replicates of each acaricide and three replicates as a negative control, immersed in distilled water. The acaricides were diluted as recommended by the manufacturers, and the ticks were submerged for five minutes. Mortality of engorged females, production of eggs, and percentage of larval hatching were evaluated. Tick mortality was 2.5%, 7.5%, and 0% for Amitraz, Cypermethrin, and Deltamethrin, respectively. The percentage of larval hatching was 53.7% for Amitraz, 88.7% for Cypermethrin, and 80.0% for Deltamethrin. As recommended by the FAO, for the acaricide to be considered effective, it must have a control rate ≥95%. Among the tested acaricides, Amitraz showed an efficacy of 90.5%, Cypermethrin 10.4%, and Deltamethrin 26.6%. Ticks from the lower Amazon showed marked resistance to Cypermethrin and Deltamethrin and mild resistance to Amitraz. This is the first report of acaricidal resistance in the region.

Pluripotency factors regulate the onset of Hox cluster activation in the early embryo.

Pluripotent cells are a transient population of the mammalian embryo dependent on transcription factors, such as OCT4 and NANOG, which maintain pluripotency while suppressing lineage specification. However, these factors are also expressed during early phases of differentiation, and their role in the transition from pluripotency to lineage specification is largely unknown. We found that pluripotency factors play a dual role in regulating key lineage specifiers, initially repressing their expression and later being required for their proper activation. We show that Oct4 is necessary for activation of HoxB genes during differentiation of embryonic stem cells and in the embryo. In addition, we show that the HoxB cluster is coordinately regulated by OCT4 binding sites located at the 3' end of the cluster. Our results show that core pluripotency factors are not limited to maintaining the precommitted epiblast but are also necessary for the proper deployment of subsequent developmental programs.

Effect of the Brazilian Cardioprotective Nutritional Program on the Quality of Life of Atherosclerotic Disease Patients

Abstract Background Cardiovascular diseases (CVD) accounted for 27% of deaths in Brazil in 2017. Most of the recorded deaths caused by CVD would be preventable if patients controlled risk factors including inadequate diet. The Brazilian Cardioprotective Nutritional Program (Dica Br) adapted the Mediterranean diet pattern to Brazilian typical foods and evaluated the effectiveness of a nutritional program based on cardioprotective foods on cardiovascular events and death of patients with cardiovascular diseases. Objectives To evaluate the effect of Dica Br on the QoL of patients with atherosclerotic disease from two health centers in the city of Rio de Janeiro. Method Randomized clinical trial with 273 participants of both sexes, over 45 years old, followed for four years. The intervention group (IG) received individualized dietary prescription, educational program, individual and group consultations, and phone calls. The control group (CG) received general dietary guidance. The SF-36 was used to assess QoL. The Student's t-test and the Mann-Whitney test was used to compare means between the groups. The mixed model test was used to compare the course of variables over time between the groups. Statistical significance was set at 5%. Result Most patients were male, with an average age of 64.2 ± 8.2 years in the IG and 65±9.5 years in the CG. Most were physically inactive, overweight, and had incomplete elementary school. The most prevalent comorbidity was systemic arterial hypertension, followed by dyslipidemia. QoL improved in both groups at four years. Waist circumference decreased in both groups over time, and low-density lipoprotein cholesterol (LDL-C) levels decreased in the IG but not in the CG after four years of follow-up. Conclusion The cardioprotective diet was effective in reducing LDL-C in the IG, and an improvement in QoL was observed in both intervention and control groups.

Blastocoel fluid removal and melatonin supplementation in the culture medium improve the viability of vitrified bovine embryos.

In this study, we investigated the effects of melatonin supplementation in the culture medium and blastocoel fluid removal (BFR) before vitrification on the quality and viability of in vitro-derived bovine embryos. After fertilization, presumptive zygotes were assigned to one of the following treatments: control, in vitro standard culture (IVC) medium; IVC + M10-9, IVC medium supplemented 10-9 M melatonin; or IVC + M10-9 BFR, IVC medium supplemented with 10-9 M melatonin plus BFR on day 7 (D7) of culture. D7 blastocysts were vitrified by the Cryotop method and, after 5 mo of storage, were warmed and incubated for an additional 72 h. The re-expansion rate was evaluated after 2 and 24 h, and the hatching rate was evaluated after 24, 48, and 72 h. At 72 h, the total number of cells (TNC); number of apoptotic cells (NAC); and expression of genes related to oxidative stress (HSPA5), cell metabolism (SLC2A3), cell repair (MSH6), placentation (KRT8 and PLAC8), and implantation (FOSL1) were assessed in the blastocysts. Less than 30% of the control blastocysts re-expanded until 2 h, whereas more than 85% of the IVC + M10-9 and IVC + M10-9 BFR blastocysts re-expanded (P < 0.05). The hatching rate of IVC + M10-9 BFR blastocysts increased at all time points (P < 0.05), reaching 66.8% at 72 h of incubation. The TNC was similar among treatments (P > 0.05), regardless of vitrification/warming and re-cultivation. The NAC:TNC was smaller for melatonin-treated blastocysts (P < 0.05). BFR increased HSPA5 (P = 0.0118) expression and did not affect SLC2A3, MSH6, KRT8, and FOSL1 expression (P > 0.05). In conclusion, melatonin (10-9 M) supplementation in the culture medium and BFR on D7 of culture increased the hatching rate 24, 48, and 72 h after warming of the vitrified embryos, indicating an improvement in cryotolerance.

Acute supplementation with beetroot juice improves endothelial function in HIV-infected individuals.

Human immunodeficiency virus (HIV) is associated with lower nitric oxide (NO) bioavailability and vascular dysfunction. Nitrate-rich beetroot juice (BJ) has been shown to acutely increase NO availability and vascular function in healthy and individuals at high risk for cardiovascular disease. Thus, we tested the effects of BJ ingestion on flow-mediated dilation (FMD) and pulse wave velocity (PWV) measurements in healthy and HIV-infected patients. Thirteen HIV-infected individuals (age, 36 ± 10 years) and 18 healthy (age, 27 ± 8 years) participated in the study. Individuals were submitted to vascular tests such as FMD and pulse PWV at pre (T0) and at 120 min (T120) after BJ and placebo (PLA) ingestion. The %FMD at T0 of the control group was significantly higher than the %FMD at T0 of the HIV individuals in both interventions. BJ improved the %FMD at T120 when compared with T0 in the HIV and control groups. There was no change in %FMD after PLA ingestion in the control and HIV groups. There were no differences between groups (control vs HIV), time points (T0 vs T120), and interventions (BJ vs PLA) for PWV. Our findings showed that nitrate-rich BJ ingestion acutely improved vascular function in healthy and HIV-infected patients. Clinical Trials Registry no. NCT03485248. Novelty: HIV is associated with lower NO bioavailability and vascular dysfunction. Acute supplementation with nitrate-rich BJ has been shown to acutely increases NO bioavailability. We showed for the first time that BJ acutely improves endothelial function in HIV-infected patients.

Dietary nitrate improves skeletal muscle microvascular oxygenation in HIV-infected patients receiving highly active antiretroviral therapy: a randomised, double-blind, cross-over, placebo-controlled study.

HIV-1 proteins and highly active antiretroviral therapy (HAART) have been associated with microvascular endothelial dysfunction. Although nitrate-rich beetroot juice (NR-BJ) consumption has been shown to improve endothelial function in clinical population, its effects in HIV-infected patients has not been addressed. We investigated the effect of a single dose of NR-BJ on muscle oxygen saturation parameters in response to a handgrip exercise in HIV-infected patients. Fifteen HIV-infected patients received NR-BJ or nitrate-depleted beetroot juice (ND-BJ) in a double-blind cross-over design. Near-IR spectroscopy was utilised to assess muscle oxygen saturation parameters during rhythmic handgrip exercise after NR-BJ or ND-BJ supplementation. A significant faster muscle oxygen desaturation rate during exercise (-7·97 (sd 5·00) v. -5·45 (3·94) %/s, P = 0·005) and muscle oxygen resaturation rate during exercise recovery (0·43 (0·24) v. 0·28 (0·24) %/s, P = 0·030) after NR-BJ ingestion was found. However, no significant difference in exercise time until fatigue was observed. Salivary nitrite and urinary nitrate concentration were analysed after NR-BJ or ND-BJ. A significant increase in salivary nitrite and urinary nitrate in NR-BJ was observed compared with ND-BJ (P < 0·05). Our findings suggest that NR-BJ consumption may acutely improve muscle oxygen saturation during exercise and exercise recovery in HIV-infected patients undergoing HAART and who are expected to present microvascular damage. Thus, future studies investigating the chronic effects of NR-BJ are warranted to delineate a better nutritional strategy based on nitrate-rich foods.

Percepções de graduandos de psicologia acerca do trabalho do psicólogo escolar e educacional / Perceptions of undergraduate psychology students about educational psychologists' work / Percepción de estudiantes de graduación en psicología acerca del trabajo del psicólogo escolar y educacional

O objetivo desta pesquisa é apreender as percepções e expectativas de estudantes de graduação em psicologia acerca da psicologia escolar e das possibilidades de intervenção de psicólogos na educação. Participaram da pesquisa 48 graduandos de psicologia de instituições de ensino superior públicas e privadas. Foram aplicados um questionário sociodemográfico e uma entrevista semiestruturada que foi registrada por meio de um gravador de voz e posteriormente analisada conforme as diretrizes apresentadas no método de Bardin. Os resultados mostraram que houve diferenças nos argumentos utilizados entre os estudantes de psicologia que haviam cursado disciplinas do campo da psicologia escolar e Educacional e aqueles que ainda não haviam cursado tais disciplinas, o que indica que uma formação que contemple disciplinas desse campo pode favorecer uma maior apropriação teórica e metodológica acerca das especificidades dessa área e das possibilidades de atuação do psicólogo na educação, em especial, seu compromisso com processos de aprendizado e desenvolvimento humano...(AU)

This study records perceptions and expectations of psychology degree students about Scholar Psychology and the possibilities of psychologist actions in Education. The sample included 48 undergraduate Psychology students of Public and Private Higher Education Institutions. A socio-demographic questionnaire and a semi-structured interview recorded via voice recorder were applied then analyzed according to the guidelines presented in Bardin's method. The results showed that there were differences in arguments used among psychology students who had taken disciplines of Scholar and Educational Psychology field and those who had not yet studied such disciplines, which indicates that training including disciplines of this field can foster greater theoretical and methodological appropriation of particularities of this area and psychologists' possibilities of acting in education, especially regarding commitment to learning and human development processes...(AU)

El objetivo de esta investigación fue identificar las percepciones y expectativas de los estudiantes de graduación en Psicología acerca de la Psicología Escolar y de las posibilidades de intervención de psicólogos en la Educación. Participaron en el estudio 48 estudiantes de Psicología de Instituciones de Enseñanza Superior Públicas y Privadas. Se aplicó un cuestionario sociodemográfico y una entrevista semiestructurada, que se ha registrado por medio de una grabadora de voz para posteriormente evaluarla según las directrices presentadas en Bardin. Los resultados mostraron que hubo diferencias en los argumentos utilizados entre los estudiantes de psicología que habían cursado asignaturas del campo de la Psicología Escolar y Educacional y aquellos que aún no las habían cursado, lo que indica que una formación que contemple asignaturas de este campo puede favorecer una mayor apropiación teórica y metodológica acerca de las especificidades de esa área y de las posibilidades de actuación del psicólogo en la educación, sobre todo, en su compromiso con procesos de aprendizaje y desarrollo humano...(AU)

Near-infrared spectroscopy-derived muscle oxygen saturation during exercise recovery and flow-mediated dilation are impaired in HIV-infected patients.

Highly active antiretroviral therapy has been associated with the presence of endothelial dysfunction in HIV-infected patients, which may impair oxygen delivery to muscles during exercise and exercise recovery. Near-infrared spectroscopy (NIRS) has been used to assess muscle oxygen saturation (SmO2) kinetics during exercise in different clinical populations in order to evaluate the balance between oxygen delivery and utilization by muscles. However, studies assessing SmO2 in HIV-infected patients have not been conducted. Therefore, the aim of the study was to evaluate NIRS-derived SmO2 during rhythmic handgrip exercise and flow-mediated dilation (FMD) in HIV-infected patients (HIV) compared to non-HIV-infected controls (N-HIV). Eighteen HIV and 17 N-HIV individuals underwent FMD assessment by ultrasound. The subjects then performed one set of rhythmic handgrip exercise until fatigue at 30% maximal isometric voluntary contraction. SmO2 was measured during entire exercise and 2-min exercise 3recovery. Muscle oxygen resaturation rate (upslope of the SmO2 over 10 s of recovery) was calculated. A significant lower FMD (3.5 ± 1.7 vs 5.9 ± 1.5%, P < 0.001) and slower oxygen resaturation rate (0.78 ± 0.4 vs 1.14 ± 0.4%·s-1, P = 0.020) in HIV as compared to N-HIV group were observed. In conclusion, our findings demonstrated that HIV-infected patients had reduced FMD and impaired muscle oxygenation during exercise recovery compared to non-HIV individuals.

Relato de Caso: paralisia unilateral de prega vocal por tuberculose mediastinal / Case report: unilateral vocal fold paralysis by mediastinal tuberculosis

Introdução: A tuberculose (TB) acomete especialmente os pulmões, no entanto, também se tem conhecimento das formas extrapulmonares. Dentre essas, buscamos relatar um caso raro de TB mediastinal localizada em trajeto de nervo laríngeo recorrente (NLR), a qual se manifestou com paralisia de prega vocal (PPV). Relato de Caso: Paciente masculino, 58 anos, apresentando quadro de disfonia. Histórico de exérese de testículo esquerdo e anatomopatológico evidenciando orquiepididimite crônica granulomatosa sugestiva de etiologia tuberculosa. Ex-tabagista e ex-etilista. Realizou, há 15 anos, um tratamento por 6 meses para TB pulmonar. Contactante de paciente bacilífero há três anos. A videolaringoscopia identificou PPV esquerda paramediana. A tomografia computadorizada (TC) de pescoço evidenciou linfonodos calcificados no mediastino superior, fossas supraclaviculares e espessamento de corda vocal à direita. A TC de Tórax apresentou conglomerados sequelares linfonodais calcificados no mediastino e sequela de processo granulomatoso crônico tipo TB em ápices pulmonares. Sorologia de HIV não reagente. Baciloscopia negativa, com amostra insuficiente. Cultura de escarro negativa. Discussão: A PPV esquerda encontrada na videolaringoscopia e TC de pescoço e os achados de TB mediastinal indicaram acometimento do NLR esquerdo pelo processo granulomatoso em seu trajeto, sendo esse, uma reativação de TB ganglionar mediastinal. Fora realizado esquema básico de tratamento para TB, com posterior resolução do quadro de disfonia. Embora a TB torácica seja uma condição comum, a rouquidão devido à PPV é uma complicação raramente associada. Conclusão: Salientamos a importância de estender a investigação ao mediastino na vigência de PPV unilateral, uma vez que a disfunção do NLR pode justificar o quadro.

Introduction: tuberculosis (TB) affects most commonly the lungs; nevertheless, extrapulmonary forms are also known. With that in mind, we intend to report a rare case of mediastinal TB located on the path of the recurrent laryngeal nerve (RLN), manifesting itself with vocal fold paralysis (VFP). Case report: male patient, 58 years old, presenting dysphonia. History of exeresis of the left testicle and anatomopathological showing chronic granulomatous orchiepididimitis, suggestive of tuberculous etiology. Former smoker, ex-alcoholic. Previous treatment for six months due to pulmonary TB, 15 years ago. Bacilli patient contactant for three years. Videolaryngoscopy identified paramedian left VFP. Computed tomography (CT) of the neck showed calcified lymph nodes in the upper mediastinum, supraclavicular fossa, and thickening on the right vocal cord. Chest CT showed calcified lymph node sequels in the mediastinal and Chronic Granulomatous TB-type sequel in pulmonary apices. Non-reactive HIV serology. Negative baciloscopy, insufficient sample. Negative sputum culture. Discussion: the finding of left VFP in videolaryngoscopy and neck CT, associated with the findings of mediastinal TB, indicated that the left RLN's involvement occurred due to the granulomatous process in its path, meaning a reactivation of mediastinal ganglionic TB. A simple treatment schedule for TB had been conducted, with subsequent resolution of the dysphonia. Even though thoracic TB is a common condition, hoarseness due to VFP is a rarely associated complication. Conclusion: we stress the importance of extending the investigation to the mediastinum in case of unilateral VFP since a dysfunction of the RLN is a possible justification of the case.

The effectiveness of temperature control of thermocyclers in PCR optimization.

This study evaluated the effectiveness of thermocycler temperature control, considering the influence of other determinant factors for the optimization of PCR. The reduction in the number of repeated PCR tests, applied in the diagnosis and prognosis of chronic myeloid leukemia at the National Cancer Institute in Brazil, was used as a measure of effectiveness. This indicator was evaluated using samples obtained before and after the temperature control in the wells of the thermocyclers. There was a reduction of 18.9% in the number of repeated exams in the second sample. A structured interview with laboratory staff indicated that there was no change in the other determinant factors.

Implementation of a Brazilian Cardioprotective Nutritional (BALANCE) Program for improvement on quality of diet and secondary prevention of cardiovascular events: A randomized, multicenter trial

Background Complex percutaneous coronary intervention (PCI) is associated with higher ischemic risk, which can be mitigated by long-term dual antiplatelet therapy (DAPT). However, concomitant high bleeding risk (HBR) may be present, making it unclear whether short- or long-term DAPT should be prioritized. Objectives This study investigated the effects of ischemic (by PCI complexity) and bleeding (by PRECISE-DAPT [PRE dicting bleeding Complications in patients undergoing stent Implantation and Sub sequent Dual Anti Platelet Therapy] score) risks on clinical outcomes and on the impact of DAPT duration after coronary stenting. Methods Complex PCI was defined as ≥3 stents implanted and/or ≥3 lesions treated, bifurcation stenting and/or stent length >60 mm, and/or chronic total occlusion revascularization. Ischemic and bleeding outcomes in high (≥25) or non-high (<25) PRECISE-DAPT strata were evaluated based on randomly allocated duration of DAPT. Results Among 14,963 patients from 8 randomized trials, 3,118 underwent complex PCI and experienced a higher rate of ischemic, but not bleeding, events. Long-term DAPT in non-HBR patients reduced ischemic events in both complex (absolute risk difference: −3.86%; 95% confidence interval: −7.71 to +0.06) and noncomplex PCI strata (absolute risk difference: −1.14%; 95% confidence interval: −2.26 to −0.02), but not among HBR patients, regardless of complex PCI features. The bleeding risk according to the Thrombolysis In Myocardial Infarction scale was increased by long-term DAPT only in HBR patients, regardless of PCI complexity. Conclusions Patients who underwent complex PCI had a higher risk of ischemic events, but benefitted from long-term DAPT only if HBR features were not present. These data suggested that when concordant, bleeding, more than ischemic risk, should inform decision-making on the duration of DAPT. (AU)

Implementation of a Brazilian Cardioprotective Nutritional (BALANCE) Program for improvement on quality of diet and secondary prevention of cardiovascular events: A randomized, multicenter trial.

BACKGROUND: Appropriate dietary recommendations represent a key part of secondary prevention in cardiovascular disease (CVD). We evaluated the effectiveness of the implementation of a nutritional program on quality of diet, cardiovascular events, and death in patients with established CVD. METHODS: In this open-label, multicenter trial conducted in 35 sites in Brazil, we randomly assigned (1:1) patients aged 45 years or older to receive either the BALANCE Program (experimental group) or conventional nutrition advice (control group). The BALANCE Program included a unique nutritional education strategy to implement recommendations from guidelines, adapted to the use of affordable and regional foods. Adherence to diet was evaluated by the modified Alternative Healthy Eating Index. The primary end point was a composite of all-cause mortality, cardiovascular death, cardiac arrest, myocardial infarction, stroke, myocardial revascularization, amputation, or hospitalization for unstable angina. Secondary end points included biochemical and anthropometric data, and blood pressure levels. RESULTS: From March 5, 2013, to Abril 7, 2015, a total of 2534 eligible patients were randomly assigned to either the BALANCE Program group (n = 1,266) or the control group (n = 1,268) and were followed up for a median of 3.5 years. In total, 235 (9.3%) participants had been lost to follow-up. After 3 years of follow-up, mean modified Alternative Healthy Eating Index (scale 0-70) was only slightly higher in the BALANCE group versus the control group (26.2 ±â€¯8.4 vs 24.7 ±â€¯8.6, P < .01), mainly due to a 0.5-serving/d greater intake of fruits and of vegetables in the BALANCE group. Primary end point events occurred in 236 participants (18.8%) in the BALANCE group and in 207 participants (16.4%) in the control group (hazard ratio, 1.15; 95% CI 0.95-1.38; P = .15). Secondary end points did not differ between groups after follow-up. CONCLUSIONS: The BALANCE Program only slightly improved adherence to a healthy diet in patients with established CVD and had no significant effect on the incidence of cardiovascular events or death.

Whey protein supplementation for the preservation of mass and muscular strength of patients with heart failure: study protocol for a randomized controlled trial.

BACKGROUND: Heart failure (HF) is an important public health problem, considered a new epidemic with high morbidity and mortality. The progression of HF often determines weight reduction, muscle mass loss, and reduced physical ability. Whey protein supplementation may increase the effects of exercise on strength and muscle mass, in addition to promoting improved endothelial function, body composition and quality of life. However, studies are needed to evaluate its benefits in patients with HF. METHODS/DESIGN: This is a double-blind, randomized, placebo-controlled clinical trial in which patients with HF will be randomly allocated to two groups to receive supplementation with whey protein or placebo, associated with supervised exercise, for 12 weeks. The frequency of exercise will be three times a week. The study variables will be evaluated at baseline and 12 weeks. The main outcome will be maintenance of muscle mass and strength. Microvascular reactivity, quality of life, and inflammatory parameters will be evaluated as secondary outcomes. DISCUSSION: HF is associated with severe loss of muscle mass and strength, directly contributing to exercise intolerance and inability to maintain daily life activities, becoming a strong predictor of reduced quality of life and mortality. The results of this study will add to the evidence base for providing new dietary recommendations. TRIAL REGISTRATION: ClinicalTrials.gov, ID: NCT03142399 . Registered on 29 May 2016. Effect of Whey Protein' Supplementation and Exercise in Patients with Heart Failure (PROT-HF).

Aging and chromatoid body assembly: Are these two physiological events linked?

The chromatoid body is a cytoplasmic male germ cell structure that plays a role in the regulation of mRNA transcription during spermatogenesis. A proteomic analysis of this structure has identified the presence of its classic molecular markers (MVH and MIWI), as well as a significant number of transient proteins. Circadian locomotor output cycles protein kaput (CLOCK) and brain and muscle ARNT-like 1 (BMAL1), which are molecular components of the circadian clock, are likely located in the chromatoid body in a transient fashion. This study sought to determine whether aging produces morphological changes in the chromatoid bodies of round spermatids similar to those previously observed in BMAL1 knockout mice. A sample of 30 male mice was divided into three groups: juvenile mice (45 days old), adult mice (120 days old), and old mice (+180 days old). Aging was confirmed by viability and sperm count analyses and testosterone dosage. Squash slides prepared with fragments of seminiferous tubules were immunostained for MVH, MIWI, BMAL1, and CLOCK detection. In juvenile and adult specimens, single round chromatoid bodies were observed using MVH/BMAL1 and MIWI/CLOCK immunostaining. In old specimens, many chromatoid bodies displayed changes in number and morphology, as well as an increase in the interactions between MVH and BMAL1; MIWI and CLOCK. Changes in chromatoid body morphology increased interactions between the proteins analyzed herein, and decreased amounts of these proteins in seminiferous tubules of older mice may indicate that aging influences the assembly and physiology of chromatoid bodies, which may, in turn, affect fertility. Impact statement The results discussed in this paper indicate that aging compromises the structure and physiology of chromatoid bodies (CBs) in post-meiotic male cells. Since CB is a fundamental structure for the differentiation of the mature male germ cell it is possible that this imbalance in CB physiology may play a role in the reduction of fertility in older men. It is important to note that not only the classic CB markers (such as the MIWI and MVH proteins) were used to showcase the structural changes in the CBs but also the main components of circadian cycle control (the CLOCK and BMAL1 proteins), indicating that the reduction of circadian control in aged males may contribute to these changes in CBs as well. Therefore, it is intriguing to evaluate the hypothesis that controlling these physiological/structural changes in CBs may be a way of delaying the effects of aging in males.

Effects of vitrification of cumulus-enclosed porcine oocytes at the germinal vesicle stage on cumulus expansion, nuclear progression and cytoplasmic maturation.

Although offspring have been produced from porcine oocytes vitrified at the germinal vesicle (GV) stage, the rate of embryo development remains low. In the present study, nuclear morphology and progression, cumulus expansion, transzonal projections (TZPs), ATP and glutathione (GSH) levels were compared between vitrified cumulus-oocyte complexes (COCs) and control COCs (no cryoprotectant treatment and no cooling), as well as a toxicity control (no cooling). Vitrification was performed with 17.5% (v/v) ethylene glycol and 17.5% (v/v) propylene glycol. Vitrification at the GV stage caused premature meiotic progression, reflected by earlier GV breakdown and untimely attainment of the MII stage. However, cytoplasmic maturation, investigated by measurement of ATP and GSH levels, as well as cumulus expansion, proceeded normally despite detectable damage to TZPs in vitrified COCs. Moreover, treatment with cryoprotectants caused fragmentation of nucleolus precursor bodies and morphological changes in F-actin from which oocytes were able to recover during subsequent IVM culture. Reduced developmental competence may be explained by premature nuclear maturation leading to oocyte aging, although other mechanisms, such as initiation of apoptosis and reduction of cytoplasmic mRNA, can also be considered. Further research will be required to clarify the presence and effects of these phenomena during the vitrification of immature COCs.