Exploring the role non-coding RNAs during myocardial cell fate.

Myocardial cell fate specification takes place during the early stages of heart development as the precardiac mesoderm is configured into two symmetrical sets of bilateral precursor cells. Molecular cues of the surrounding tissues specify and subsequently determine the early cardiomyocytes, that finally matured as the heart is completed at early postnatal stages. Over the last decade, we have greatly enhanced our understanding of the transcriptional regulation of cardiac development and thus of myocardial cell fate. The recent discovery of a novel layer of gene regulation by non-coding RNAs has flourished their implication in epigenetic, transcriptional and post-transcriptional regulation of cardiac development. In this review, we revised the current state-of-the-art knowledge on the functional role of non-coding RNAs during myocardial cell fate.

Effects of a ketogenic diet on motor function and motor unit number estimation in aged C57BL/6 mice.

OBJECTIVE: Pathological, age-related loss of muscle function, commonly referred to as sarcopenia, contributes to loss of mobility, impaired independence, as well as increased risk of adverse health events. Sarcopenia has been attributed to changes in both neural and muscular integrity during aging. Current treatment options are primarily limited to exercise and dietary protein fortification, but the therapeutic impact of these approaches are often inadequate. Prior work has suggested that a ketogenic diet (KD) might improve healthspan and lifespan in aging mice. Thus, we sought to investigate the effects of a KD on neuromuscular indices of sarcopenia in aged C57BL/6 mice. DESIGN: A randomized, controlled pre-clinical experiment consisting of longitudinal assessments performed starting at 22-months of age (baseline) as well as 2, 6 and 10 weeks after the start of a KD vs. regular chow intervention. SETTING: Preclinical laboratory study. SAMPLE SIZE: Thirty-six 22-month-old mice were randomized into 2 dietary groups: KD [n = 22 (13 female and 9 male)], and regular chow [n = 15 (7 female and 8 male)]. MEASUREMENTS: Measures included body mass, hindlimb and all limb grip strength, rotarod for motor performance, plantarflexion muscle contractility, motor unit number estimations (MUNE), and repetitive nerve stimulation (RNS) as an index of neuromuscular junction transmission efficacy recorded from the gastrocnemius muscle. At end point, muscle wet weight and blood samples were collected to assess blood beta-hydroxybutyrate levels. STATISTICAL ANALYSIS: Primary analyses were two-way mixed effects ANOVA (diet and time × diet) to determine the effect of a KD on indices of motor function (grip, rotarod) and indices of motor unit (MUNE) and muscle (contractility) function. RESULTS: Beta-hydroxybutyrate (BHB) was significantly higher at 10 weeks in mice on a KD vs control group (0.83 ± 0.44 mmol/l versus 0.42 ± 0.21 mmol/l, η2 = 0.265, unpaired t-test, p = 0.0060). Mice on the KD intervention demonstrated significantly increased hindlimb grip strength (diet, p = 0.0001; time × diet, p = 0.0030), all limb grip strength (diet, p = 0.0005; time × diet, p = 0.0523), and rotarod latency to fall (diet, p = 0.0126; time × diet, p = 0.0021). Mice treated with the KD intervention also demonstrated increased MUNE (diet, p = 0.0465; time × diet, p = 0.0064), but no difference in muscle contractility (diet, p = 0.5248; time × diet, p = 0.5836) or RNS (diet, p = 0.3562; time × diet, p = 0.9871). CONCLUSION: KD intervention improved neuromuscular and motor function in aged mice. This pre-clinical work suggests that further research is needed to assess the efficacy and physiological effects of a KD on indices of sarcopenia.

Novel Technique to Increase the Effective Workspace of a Soft Robot.

This article presents a novel technique for a class 2 tensegrity robot, also classified as a soft robot, to increase workspace by increasing the number of geometric equilibrium configurations of the robot. The proposed modification, unlike the strategies reported in the literature, consists of increasing the number of points where the flexible and rigid elements that make up the robot come into contact without the need to increase the number of actuators, the number of flexible elements, or modify the geometry of the rigid elements. The form-finding methodology combines the basic principles of statics with the direct and inverse kinematic position analysis to determine the number of equilibrium positions of the modified robot. In addition, numerical experiments were carried out using the commercial software ANSYS®, R18.2 based on the finite element theory, to corroborate the results obtained with them. With the proposed modification, an increase of 23.369% in the number of geometric equilibrium configurations is achieved, which integrates the workspace of the modified class 2 tensegrity robot. The novel technique applied to tensegrity robots and the tools developed to increase their workspace apply perfectly to scale the robots presented in this paper.

De-escalating treatment indications for patients who achieve metabolic goals.

INTRODUCTION: Robust evidence exists regarding initiation, intensification or modification of treatments. Recommendations to de-escalate therapy are lacking, specifically in diabetes. A successful treatment de-intensification reduces overtreatment, polypharmacy, and risk of adverse effects. OBJECTIVE: To encompass current recommendations for deprescribing common drugs and create a consensus among health professionals. METHODS: We reviewed four databases for deprescribing approaches published between 2010 and 2022. Articles were divided into different groups of drugs (for uric-acid, hypoglycemic, lipid-lowering, and psychotropic drugs). RESULTS: Hypoglycemic agents: strategies were limited to newer agents and insulin regimens for elderly individuals. Reducing insulin was associated with 1.1% reduction of A1c over time. SGLT2i and GLP-1RAs dose reduction depends on adverse events. Lipid-lowering agents: studies show that patients with very low cholesterol have fewer cardiovascular events without associated increased risk. Antihypertensive agents: Younger patients, lower systolic blood pressure, and few comorbidities are ideal characteristics for discontinuation. Uric acid therapy: we found no recommendation for dose de-escalation. Poor treatment adherence is associated with episodes of gout and deforming arthritis in the long term. CONCLUSION: Deprescribing hypoglycemic, statins, antihypertensives, and urate-lowering agents may be feasible in selected patients, but periodic surveillance is important. More evidence is necessary to support this decision entirely.

miR-195b is required for proper cellular homeostasis in the elderly.

Over the last decade we have witnessed an increasing number of studies revealing the functional role of non-coding RNAs in a multitude of biological processes, including cellular homeostasis, proliferation and differentiation. Impaired expression of non-coding RNAs can cause distinct pathological conditions, including herein those affecting the gastrointestinal and cardiorespiratory systems, respectively. miR-15/miR-16/miR-195 family members have been broadly implicated in multiple biological processes, including regulation of cell proliferation, apoptosis and metabolism within distinct tissues, such as heart, liver and lungs. While the functional contribution of miR-195a has been reported in multiple biological contexts, the role of miR-195b remains unexplored. In this study we dissected the functional role of miR-195b by generating CRISPR-Cas9 gene edited miR-195b deficient mice. Our results demonstrate that miR-195b is dispensable for embryonic development. miR-195b-/- mice are fertile and displayed no gross anatomical and/or morphological defects. Mechanistically, cell cycle regulation, metabolism and oxidative stress markers are distinctly impaired in the heart, liver and lungs of aged mice, a condition that is not overtly observed at midlife. The lack of overt functional disarray during embryonic development and early adulthood might be due to temporal and tissue-specific compensatory mechanisms driven by selective upregulation miR-15/miR-16/miR-195 family members. Overall, our data demonstrated that miR-195b is dispensable for embryonic development and adulthood but is required for cellular homeostasis in the elderly.

Ketogenic Diet Improves Motor Function and Motor Unit Connectivity in Aged C57BL/6 Mice.

Objective: Pathological, age-related loss of muscle function, commonly referred to as sarcopenia, contributes to loss of mobility, impaired independence, as well as increased risk of adverse health events. Sarcopenia has been attributed to changes in both neural and muscular integrity during aging. Current treatment options are primarily limited to exercise and dietary protein fortification, but the therapeutic impact of these approaches are often inadequate. Prior work has suggested that a ketogenic diet (KD) might improve healthspan and lifespan in aging mice. Thus, we sought to investigate the effects of a KD on neuromuscular indices of sarcopenia in aged C57BL/6 mice. Design: A randomized, controlled pre-clinical experiment consisting of longitudinal assessments performed starting at 22-months of age (baseline) as well as 2, 6 and 10 weeks after the start of a KD vs. regular chow intervention. Setting: Preclinical laboratory study. Sample size: Thirty-six 22-month-old mice were randomized into 2 dietary groups: KD [n = 22 (13 female and 9 male)], and regular chow [n = 15 (7 female and 8 male)]. Measurements: Measures included body mass, hindlimb and all limb grip strength, rotarod for motor performance, plantarflexion muscle contractility, motor unit number estimations (MUNE), and repetitive nerve stimulation (RNS) as an index of neuromuscular junction transmission efficacy recorded from the gastrocnemius muscle. At end point, blood samples were collected to assess blood beta-hydroxybutyrate levels. Statistical Analysis: Two-way ANOVA mixed-effects analysis (time x diet) were performed to analyze grip, rotarod, MUNE, and muscle contractility data. Results: Beta-hydroxybutyrate (BHB) was significantly higher at 10 weeks in mice on a KD vs control group (0.83 ± 0.44 mmol/l versus 0.42 ± 0.21 mmol/l, η2 = 0.265, unpaired t-test, p = 0.0060). Mice on the KD intervention demonstrated significantly increased hindlimb grip strength (time x diet, p = 0.0030), all limb grip strength (time x diet, p = 0.0523), and rotarod latency to fall (time x diet, p = 0.0021). Mice treated with the KD intervention also demonstrated significantly greater MUNE (time x diet, p = 0.0064), but no difference in muscle contractility (time x diet, p = 0.5836) or RNS (time x diet, p = 0.9871). Conclusion: KD intervention improved neuromuscular and motor function in aged mice. This pre-clinical work suggests that further research is needed to assess the efficacy and physiological effects of a KD on indices of sarcopenia.

Neutralization of SARS-CoV-2 (lineage B.1.1) by hyperimmune llama (Lama glama) serum in vero cell culture. / Neutralización del virus SARS-CoV-2 (linaje B.1.1) por suero hiperinmune de llama (Lama glama) en cultivo de células Vero.

OBJECTIVE.: To evaluate the serological antibody response of a llama (Lama glama) to SARS-CoV-2 (B.1.1 lineage) immunization and the neutralizing capacity of hyperimmune llama serum against SARS-CoV-2 virus (B.1.1 lineage) in Vero cells. MATERIALS AND METHODS.: A llama was immunized with inactivated SARS-CoV-2 (B.1.1 lineage). Serum samples were analyzed to evaluate the level of antibodies by ELISA, as well as reactivity to SARS-CoV-2 antigens by Western Blot. In addition, viral neutralization in cell cultures was assessed by the Plate Reduction Neutralization Test (PRNT). RESULTS: . Seroreactivity increased in the immunized llama from week 4 onwards. Antibody titers were the highest after the seventh immunization booster. Western blot results confirmed the positive ELISA findings, and immune serum antibodies recognized several viral proteins. The neutralization assay (PRNT) showed visible viral neutralization, which was in accordance with the ELISA and Western Blot results. CONCLUSIONS.: The findings suggest that hyperimmune llama serum could constitute a source of therapeutic antibodies against SARS-CoV-2 infections (lineage B.1.1), and should be studied in further research.

OBJETIVO.: Evaluar la respuesta serológica de anticuerpos de una llama (Lama glama) a la inmunización del virus SARS-CoV-2 (linaje B.1.1) y la capacidad neutralizante del suero de llama hiperinmune frente al virus SARS-CoV-2 (linaje B.1.1) en células Vero. MATERIALES Y MÉTODOS.: Se inmunizó una llama con el virus SARS-CoV-2 inactivado (Linaje B.1.1) y se analizaron muestras de suero para evaluar el nivel de anticuerpos mediante ELISA, así como la reactividad a antígenos de SARS-CoV-2 mediante Western Blot. Además, se evaluó la neutralización viral en cultivos celulares por la Prueba de Neutralización por Reducción de Placas (PRNT, por sus siglas en inglés). RESULTADOS.: Se observó un aumento en la serorreactividad en la llama inmunizada desde la semana 4 en adelante. Los títulos de anticuerpos fueron más elevados en el séptimo refuerzo de inmunización. Los resultados de Western Blot confirmaron los hallazgos positivos del ELISA, y los anticuerpos del suero inmune reconocieron varias proteínas virales. El ensayo de neutralización (PRNT) mostró una neutralización viral visible, concordante con los resultados de ELISA y Western Blot. CONCLUSIONES.: Los hallazgos sugieren que el suero hiperinmune de llama podría constituir una fuente de anticuerpos terapéuticos contra las infecciones por el virus SARS-CoV-2 (linaje B.1.1) y que deberá ser evaluado en estudios posteriores.

LncRNAs and CircRNAs in Endoplasmic Reticulum Stress: A Promising Target for Cardiovascular Disease?

The endoplasmic reticulum (ER) is a principal subcellular organelle responsible for protein quality control in the secretory pathway, preventing protein misfolding and aggregation. Failure of protein quality control in the ER triggers several molecular mechanisms such as ER-associated degradation (ERAD), the unfolded protein response (UPR) or reticulophagy, which are activated upon ER stress (ERS) to re-establish protein homeostasis by transcriptionally and translationally regulated complex signalling pathways. However, maintenance over time of ERS leads to apoptosis if such stress cannot be alleviated. The presence of abnormal protein aggregates results in loss of cardiomyocyte protein homeostasis, which in turn results in several cardiovascular diseases such as dilated cardiomyopathy (DCM) or myocardial infarction (MI). The influence of a non-coding genome in the maintenance of proper cardiomyocyte homeostasis has been widely proven. To date, the impact of microRNAs in molecular mechanisms orchestrating ER stress response has been widely described. However, the role of long noncoding RNAs (lncRNAs) and circular RNAs (circRNAs) is just beginning to be addressed given the potential role of these RNA classes as therapeutic molecules. Here, we provide a current state-of-the-art review of the roles of distinct lncRNAs and circRNAs in the modulation of ERS and UPR and their impact in cardiovascular diseases.

State of Charge Estimation Model Based on Genetic Algorithms and Multivariate Linear Regression with Applications in Electric Vehicles.

Nowadays, the use of renewable, green/eco-friendly technologies is attracting the attention of researchers, with a view to overcoming recent challenges that must be faced to guarantee the availability of Electric Vehicles (EVs). Therefore, this work proposes a methodology based on Genetic Algorithms (GA) and multivariate regression for estimating and modeling the State of Charge (SOC) in Electric Vehicles. Indeed, the proposal considers the continuous monitoring of six load-related variables that have an influence on the SOC (State of Charge), specifically, the vehicle acceleration, vehicle speed, battery bank temperature, motor RPM, motor current, and motor temperature. Thus, these measurements are evaluated in a structure comprised of a Genetic Algorithm and a multivariate regression model in order to find those relevant signals that better model the State of Charge, as well as the Root Mean Square Error (RMSE). The proposed approach is validated under a real set of data acquired from a self-assembly Electric Vehicle, and the obtained results show a maximum accuracy of approximately 95.5%; thus, this proposed method can be applied as a reliable diagnostic tool in the automotive industry.

Re-estimation of juvenile Isurus oxyrinchus growth in the Mexican Pacific through a multimodel inference approach and verification of growth band periodicity.

An update of the age and growth for juveniles of the short fin mako shark (I. oxyrinchus) from the Mexican Pacific is presented, based on the analysis of growth band counts from dorsal vertebrae of 198 individuals [110 females, 74-231 cm of total length (TL) and 88 males, 72-231 cm TL) caught during 2008-2018. New available information on vertebral growth band periodicity (biannual deposition in juveniles) and the convenience of using vertebrae form the dorsal region over the cervical region to count growth bands, as well as a multimodel approach, were used. The von Bertalanffy (VB) growth model, Gompertz, logistic and two parameters of VB (2-VB) were fitted to the length-at-age. Only ages ≤6 years were used for the fitting of the models and their performance was compared with the small-sample bias-corrected form of the Akaike information criterion (AICc), their differences ( ∆ i ) and weights ( w i ). Following a multimodel inference approach, the model averaged asymptotic length ( L ¯ inf ), length-at-age 0 ( L ¯ 0 ) and their unconditional standard error ( SE ¯ ), were estimated for each sex scenario using the three-parameter version of each model. The precision of growth band counts was acceptable for the different methods used and by two different readers. The centrum edge analysis (CEA) and marginal increment analysis (MIA) did not support the hypothesis of biannual band pair formation for juveniles, likewise for adults the periodicity could not be verified due to the small sample of large animals. Age was estimated assuming the formation of two pairs of growth bands per year during the first 5 years and one pair of bands per year afterwards considering direct validation information. The estimated ages in years ranged from 0-14 for females and 0-6 for males. The Kimura likelihood ratio test showed no differences in the growth curves of juveniles by sex (P > 0.05). According to the AICc, the 2-VB model better fitted the length-at-age data for combined sexes (Linf = 386.4 cm, k = 0.12 years-1 , L0 = 70 cm). The model averaged L ¯ inf and L ¯ 0 were 378.3 cm ( SE ¯ = 64.5 ) and 69.5 cm ( SE ¯ = 6.3 ), respectively. The growth parameters determined for juveniles of I. oxyrinchus are similar to those estimated in other regions, showing relatively fast growth rate as previously reported, medium longevity in comparison to other shark species and natural mortality close to that reported in the last stock assessment for the North Pacific Ocean. These life-history parameters should be considered to evaluate the population in the region and to develop better fishery management and conservation measures.

Comparative Analysis of Non-Coding RNA Transcriptomics in Heart Failure.

Heart failure constitutes a clinical complex syndrome with different symptomatic characteristics depending on age, sex, race and ethnicity, among others, which has become a major public health issue with an increasing prevalence. One of the most interesting tools seeking to improve prevention, diagnosis, treatment and prognosis of this pathology has focused on finding new molecular biomarkers since heart failure relies on deficient cardiac homeostasis, which is regulated by a strict gene expression. Therefore, currently, analyses of non-coding RNA transcriptomics have been oriented towards human samples. The present review develops a comparative study emphasizing the relevance of microRNAs, long non-coding RNAs and circular RNAs as potential biomarkers in heart failure. Significantly, further studies in this field of research are fundamental to supporting their widespread clinical use. In this sense, the various methodologies used by the authors should be standardized, including larger cohorts, homogeneity of the samples and uniformity of the bioinformatic pipelines used to reach stratification and statistical significance of the results. These basic adjustments could provide promising steps to designing novel strategies for clinical management of patients with heart failure.

Corporoplasty under local anesthesia: Cost-effective improvement of the surgical treatment of Peyronie's disease / Corporoplastia bajo anestesia local: mejora del coste-efectividad del tratamiento quirúrgico de la enfermedad de Peyronie

Introduction and objectives: Peyronie's disease produces penile curvature that leads, in certain cases, to difficulties in having satisfactory sexual relations. The objective of this study is to evaluate the functional and cost-effectiveness results obtained in the surgical treatment of penile curvature due to Peyronie's disease under local anesthesia, comparing them with those performed under a general–spinal anesthesia regimen. Material and methods: Two groups of patients undergoing corporoplasty are compared according to the type of anesthesia used. Group 1 consists of 32 patients who underwent major outpatient surgery and under local anesthesia from June 2016 to June 2019. Their data are collected prospectively. Group 2 consists of 30 patients who underwent surgery under general/spinal anesthesia with hospital admission, from January 2013 to December 2015, with their data collected retrospectively. Anesthetic and surgical procedure, postoperative functional results, degree of satisfaction and hospital costs between both groups are analyzed, considering p≤0.05 as statistical significance and analyzing the results with the SPSS 20.0 program. Results: Of the 32 patients included in group 1, none required admission for intra or postoperative complications. In both groups, an improvement of the PDQ-test was observed without statistically significant differences, with the degree of global satisfaction above 95% in both groups. There were also no differences in the appearance of complications secondary to the anesthetic procedure or the hospital regime. We observed differences in hospital costs, being 44% lower for the group performed with local anesthesia. (AU)

Introducción y objetivos: La enfermedad de Peyronie produce una curvatura del pene que conduce, en ciertos casos, a dificultades para tener relaciones sexuales satisfactorias. El objetivo de este estudio es evaluar los resultados funcionales y de coste-efectividad obtenidos en el tratamiento quirúrgico de la curvatura peneana por enfermedad de Peyronie bajo anestesia local, comparándolos con los conseguidos bajo un régimen de anestesia general-raquídea. Material y métodos: Se comparan 2 grupos de pacientes sometidos a corporoplastia según el tipo de anestesia utilizada. El grupo 1 está formado por 32 pacientes que se sometieron a cirugía mayor ambulatoria y bajo anestesia local desde junio de 2016 a junio de 2019. Sus datos se recogen de forma prospectiva. El grupo 2 está formado por 30 pacientes intervenidos bajo anestesia general/raquídea con ingreso hospitalario, desde enero de 2013 hasta diciembre de 2015, con sus datos recogidos de forma retrospectiva. Se analiza el procedimiento anestésico y quirúrgico, los resultados funcionales postoperatorios, el grado de satisfacción y los costos hospitalarios entre ambos grupos, considerando p≤0,05 como significación estadística y analizando los resultados con el programa SPSS® 20.0. Resultados: De los 32 pacientes incluidos en el grupo 1, ninguno requirió ingreso por complicaciones intra o postoperatorias. En ambos grupos se observó una mejora en el test PDQ sin diferencias estadísticamente significativas, con un grado de satisfacción global superior al 95% en ambos grupos. Tampoco hubo diferencias en la aparición de complicaciones secundarias al procedimiento anestésico o al régimen hospitalario. Observamos diferencias en los costos hospitalarios, siendo un 44% menor para el grupo en el que se utilizó anestesia local. (AU)

LncRNA H19 Impairs Chemo and Radiotherapy in Tumorigenesis.

Various treatments based on drug administration and radiotherapy have been devoted to preventing, palliating, and defeating cancer, showing high efficiency against the progression of this disease. Recently, in this process, malignant cells have been found which are capable of triggering specific molecular mechanisms against current treatments, with negative consequences in the prognosis of the disease. It is therefore fundamental to understand the underlying mechanisms, including the genes-and their signaling pathway regulators-involved in the process, in order to fight tumor cells. Long non-coding RNAs, H19 in particular, have been revealed as powerful protective factors in various types of cancer. However, they have also evidenced their oncogenic role in multiple carcinomas, enhancing tumor cell proliferation, migration, and invasion. In this review, we analyze the role of lncRNA H19 impairing chemo and radiotherapy in tumorigenesis, including breast cancer, lung adenocarcinoma, glioma, and colorectal carcinoma.

New Insights into the Roles of lncRNAs as Modulators of Cytoskeleton Architecture and Their Implications in Cellular Homeostasis and in Tumorigenesis.

The importance of the cytoskeleton not only in cell architecture but also as a pivotal element in the transduction of signals that mediate multiple biological processes has recently been highlighted. Broadly, the cytoskeleton consists of three types of structural proteins: (1) actin filaments, involved in establishing and maintaining cell shape and movement; (2) microtubules, necessary to support the different organelles and distribution of chromosomes during cell cycle; and (3) intermediate filaments, which have a mainly structural function showing specificity for the cell type where they are expressed. Interaction between these protein structures is essential for the cytoskeletal mesh to be functional. Furthermore, the cytoskeleton is subject to intense spatio-temporal regulation mediated by the assembly and disassembly of its components. Loss of cytoskeleton homeostasis and integrity of cell focal adhesion are hallmarks of several cancer types. Recently, many reports have pointed out that lncRNAs could be critical mediators in cellular homeostasis controlling dynamic structure and stability of the network formed by cytoskeletal structures, specifically in different types of carcinomas. In this review, we summarize current information available about the roles of lncRNAs as modulators of actin dependent cytoskeleton and their impact on cancer pathogenesis. Finally, we explore other examples of cytoskeletal lncRNAs currently unrelated to tumorigenesis, to illustrate knowledge about them.

Removal of Ochratoxin A from Red Wine Using Alginate-PVA-L. plantarum (APLP) Complexes: A Preliminary Study.

The presence of ochratoxin A (OTA) in wines is a problem mainly due to the health damage it can cause to frequent drinkers. A method for removing these toxic substances from wine is the use of lactic acid bacteria with mycotoxin-adsorption capacities; however, their use is limited since a matrix in which they can be immobilized, to remove them after use, is needed. In this study, L. plantarum (LP) was encapsulated in a polymeric matrix composed of polyvinyl alcohol (PVA) and alginate, forming alginate-PVA-LP (APLP) complexes. Then, these complexes were characterized, and assays of OTA and phenol removal from wines were performed. As a result, it was observed that the APLP complexes at a concentration of 0.5 g mL-1 removed over 50% of the OTA without substantially affecting the concentration of total phenols. In addition, it was determined that the presence of L. plantarum directly affected the ability to adsorb OTA from wines and did not decrease the total phenols. In conclusion, an alginate-PVA matrix allows immobilizing LP, and the complexes formed are an alternative for removing ochratoxin from contaminated wines.

Inhibition of RhoA and Cdc42 by miR-133a Modulates Retinoic Acid Signalling during Early Development of Posterior Cardiac Tube Segment.

It is well known that multiple microRNAs play crucial roles in cardiovascular development, including miR-133a. Additionally, retinoic acid regulates atrial marker expression. In order to analyse the role of miR-133a as a modulator of retinoic acid signalling during the posterior segment of heart tube formation, we performed functional experiments with miR-133a and retinoic acid by means of microinjections into the posterior cardiac precursors of both primitive endocardial tubes in chick embryos. Subsequently, we subjected embryos to whole mount in situ hybridisation, immunohistochemistry and qPCR analysis. Our results demonstrate that miR-133a represses RhoA and Cdc42, as well as Raldh2/Aldh1a2, and the specific atrial markers Tbx5 and AMHC1, which play a key role during differentiation. Furthermore, we observed that miR-133a upregulates p21 and downregulates cyclin A by repressing RhoA and Cdc42, respectively, thus functioning as a cell proliferation inhibitor. Additionally, retinoic acid represses miR-133a, while it increases Raldh2, Tbx5 and AMHC1. Given that RhoA and Cdc42 are involved in Raldh2 expression and that they are modulated by miR-133a, which is influenced by retinoic acid signalling, our results suggest the presence of a negative feedback mechanism between miR-133a and retinoic acid during early development of the posterior cardiac tube segment. Despite additional unexplored factors being possible contributors to this negative feedback mechanism, miR-133a might also be considered as a potential therapeutic tool for the diagnosis, therapy and prognosis of cardiac diseases.

Post-Transcriptional Regulation of Molecular Determinants during Cardiogenesis.

Cardiovascular development is initiated soon after gastrulation as bilateral precardiac mesoderm is progressively symmetrically determined at both sides of the developing embryo. The precardiac mesoderm subsequently fused at the embryonic midline constituting an embryonic linear heart tube. As development progress, the embryonic heart displays the first sign of left-right asymmetric morphology by the invariably rightward looping of the initial heart tube and prospective embryonic ventricular and atrial chambers emerged. As cardiac development progresses, the atrial and ventricular chambers enlarged and distinct left and right compartments emerge as consequence of the formation of the interatrial and interventricular septa, respectively. The last steps of cardiac morphogenesis are represented by the completion of atrial and ventricular septation, resulting in the configuration of a double circuitry with distinct systemic and pulmonary chambers, each of them with distinct inlets and outlets connections. Over the last decade, our understanding of the contribution of multiple growth factor signaling cascades such as Tgf-beta, Bmp and Wnt signaling as well as of transcriptional regulators to cardiac morphogenesis have greatly enlarged. Recently, a novel layer of complexity has emerged with the discovery of non-coding RNAs, particularly microRNAs and lncRNAs. Herein, we provide a state-of-the-art review of the contribution of non-coding RNAs during cardiac development. microRNAs and lncRNAs have been reported to functional modulate all stages of cardiac morphogenesis, spanning from lateral plate mesoderm formation to outflow tract septation, by modulating major growth factor signaling pathways as well as those transcriptional regulators involved in cardiac development.