MicroRNA-20a-5p Downregulation by Atorvastatin: A Potential Mechanism Involved in Lipid-Lowering Therapy.

The treatment of hypercholesterolemia is mainly based on statins. However, the response to pharmacological therapy shows high inter-individual variability, resulting in variable effects in both lipid lowering and risk reduction. Thus, a better understanding of the lipid-lowering mechanisms and response variability at the molecular level is required. Previously, we demonstrated a deregulation of the microRNA expression profile in HepG2 cells treated for 24 h with atorvastatin, using a microarray platform. In the present study, we evaluated the expression of hsa-miR-17-5p, hsa-miR-20a-5p and hsa-miR-106a-5p in hypercholesterolemic patients before and after atorvastatin treatment and in HepG2 cells treated for 24 h with atorvastatin The miRNA hsa-mir-20a-5p was repressed after atorvastatin treatment in hypercholesteremic subjects and in HepG2 cells in culture. Repression of hsa-mir-20a-5p increased LDLR gene and protein expression in HepG2 cells, while hsa-mir-20a-5p overexpression reduced LDLR gene and protein expression.

Physical activity level and physical fitness in subjects with chronic musculoskeletal pain: a cross-sectional study.

Background: Low physical activity (PA) levels and low physical fitness (PF) have been reported in subjects with temporality-based chronic pain; however, it is unknown whether there are differences in subjects with nociplastic pain (NP) compared with subjects with non-nociplastic pain (NNP). Objective: The aim was to compare the levels of PA and PF in patients with chronic, nociplastic, and non-nociplastic musculoskeletal pain. Methods: This is an analytical, cross-sectional study. The sample comprised 30 patients receiving ambulatory physiotherapy treatment. Pain was classified as NP or NNP according to the International Association for the Study of Pain categorization system. The PA level was measured with the International Physical Activity Questionnaire-Short Form, and the PF level was measured with the hand grip strength test (HGS) to assess upper limb strength, the five Repetition Sit-to-Stand Test (5R-SRTS) to assess lower limb strength and power, and the YMCA 3 Min Step Test (YMCA-3MST) to estimate peak VO2. The results were compared with independent samples t-tests (with p < 0.05 considered significant). Cohen's d was calculated to determine the effect size. Results: The NP group reported a significantly lower PA level than the NNP group, specifically the vigorous PA (p = 0.0009), moderate PA (p = 0.0002), and total PA (p = 0.005) dimensions. The NP group also showed significantly lower 5R-STS (p = 0.000) and HGS (p = 0.002) results compared with the NNP group. There were no significant differences in the YMCA-3MST between the NP and NNP groups (p = 0.635). Conclusion: It is possible that the neurophysiological and neuromuscular changes related to NP are associated with a reduced ability to perform vigorous PA. Clinicians should identify the presence of NP comorbidities in conjunction with the diagnosis when establishing the therapeutic goals.

Modulation of the Cardiovascular Effects of Polycyclic Aromatic Hydrocarbons: Physical Exercise as a Protective Strategy.

Exposure to polycyclic aromatic hydrocarbons (PAHs) present in air pollution increases cardiovascular risk. On the contrary, physical exercise is a widely used therapeutic approach to mitigate cardiovascular risk, but its efficacy in an environment of air pollution, particularly with PAHs, remains unclear. This study investigates the effects of exercise on inflammation, endothelial dysfunction, and REDOX imbalance due to PAH exposure using a mouse model. Twenty male BALB/c mice were subjected to a mixture of PAHs (phenanthrene, fluoranthene, pyrene) in conjunction with aerobic exercise. The investigation evaluated serum levels of inflammatory cytokines, gene expression linked to inflammatory markers, endothelial dysfunction, and REDOX imbalance in aortic tissues. Furthermore, the study evaluated the expression of the ICAM-1 and VCAM-1 proteins. Exercise led to notable changes in serum inflammatory cytokines, as well as the modulation of genes associated with endothelial dysfunction and REDOX imbalance in aortic tissue. In turn, exercise produced a modulation in the protein expression of ICAM-1 and VCAM-1. The findings implicate the potential of exercise to counter PAH-induced damage, as demonstrated by changes in markers. In conclusion, exercise could mitigate the adverse effects related to exposure to PAHs present in air pollution, as evidenced by changes in inflammatory markers, endothelial dysfunction, and REDOX imbalance.

Polycyclic Aromatic Hydrocarbons (PAHs) Exposure Triggers Inflammation and Endothelial Dysfunction in BALB/c Mice: A Pilot Study.

The particulate matter present in air pollution is a complex mixture of solid and liquid particles that vary in size, origin, and composition, among which are polycyclic aromatic hydrocarbons (PAHs). Although exposure to PAHs has become an important risk factor for cardiovascular disease, the mechanisms by which these compounds contribute to increased cardiovascular risk have not been fully explored. The aim of the present study was to evaluate the effects of PAH exposure on systemic pro-inflammatory cytokines and markers of endothelial dysfunction. An intervention was designed using a murine model composed of twenty BALB/c male mice separated into controls and three groups exposed to a mixture of phenanthrene, fluoranthene, and pyrene using three different concentrations. The serum levels of the inflammatory cytokines and gene expression of adhesion molecules located on endothelial cells along with inflammatory markers related to PAH exposure in aortic tissue were determined. Furthermore, the expression of the ICAM-1 and VCAM-1 proteins was evaluated. The data showed significant differences in IL-6 and IFN-γ in the serum. In the gene expression, significant differences for ICAM-1, VCAM-1, and E-Selectin were observed. The results suggest that phenanthrene, fluoranthene, and pyrene, present in air pollution, stimulate the increase in serum inflammatory cytokines and the expression of markers of endothelial dysfunction in the murine model studied, both relevant characteristics associated with the onset of disease atherosclerosis and cardiovascular disease.

Attitude towards Older People According to Sociodemographic and Educational Variables in Students of a Chilean University.

Current evidence suggests that attitude towards older people may be associated with sociodemographic and educational variables; hence, a positive attitude towards older people is key when training new university professionals. However, there is little evidence of this association in Chilean university students. The objective was to analyze students from a Chilean university's attitudes towards older people, according to sociodemographic and educational variables. Analytical and cross-sectional study; 515 students from a Chilean university were consulted online about their attitude towards older people using Kogan's Attitudes towards Old People scale. Additionally, sociodemographic and educational variables were recorded. The average score for positive attitude was 70.8 (±9.7), while the negative attitude score was 68.3 (±11.6). The total score was 139.1 (±16.6). Mostly, university students perceive themselves with a low-level positive attitude (61.2%). Additionally, older university students (26-42 years old); women; Chileans; students of law, speech therapy, and occupational therapy; students in their final years of the programs; and those who had training in older people outside the university have a more positive attitude towards older people. In Conclusion, a profile of sociodemographic and educational characteristics of students with a lower and higher attitude towards older age was investigated. These results are relevant since the way of seeing the aging process could regulate the training of future professionals and consequently generate changes in dealing with older people. Young people's perception of ageing would affect the treatment and incorporation of the older people in society and the adaptation of policies in this age group.

Characterization of microbial communities from gut microbiota of hypercholesterolemic and control subjects.

Introduction: In recent years, several studies have evidenced the importance of the microbiome to host physiology as metabolism regulator, along with its potential role in triggering various diseases. In this study, we analyzed the gut microbiota in hypercholesterolemic (cases) and normocholesterolemic (controls) individuals to identify characteristic microbial signature for each condition. Methods: Stool samples were obtained from 57 adult volunteers (27 hypercholesterolemic and 30 controls). The taxonomic profiling of microbial communities was performed using high-throughput sequencing of 16S rRNA V3-V4 amplicons, followed by data analysis using Quantitative Insights Into Microbial Ecology 2 (QIIME2) and linear discriminant analysis (LDA) effect size (LEfSe). Results: Significant differences were observed in weight, height, body mass index (BMI) and serum levels of triglycerides, total cholesterol and low-density lipoprotein cholesterol (LDL-C) between the groups (p<0.05). LEfSe showed differentially abundant prokaryotic taxa (α=0.05, LDA score > 2.0) in the group of hypercholesterolemic individuals (Methanosphaera, Rothia, Chromatiales, Clostridiales, Bacillaceae and Coriobacteriaceae) and controls (Faecalibacterium, Victivallis and Selenomonas) at various taxonomic levels. In addition, through the application of Phylogenetic Investigation of Communities by Reconstruction of Unobserved States 2 (PICRUSt2), the predominance of pathways related to biosynthesis in hypercholesterolemic patients was established, compared to controls in which degradation pathways were predominant. Finally, in the analysis of co-occurrence networks, it was possible to identify associations between the microorganisms present in both studied groups. Conclusion: Our results point out to unique microbial signatures, which likely play a role on the cholesterol metabolism in the studied population.

Unlocking SARS-CoV-2 detection in low- and middle-income countries.

Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes and nucleic acids can supply over 50,000 tests per production batch. The optimized one-step RT-PCR coupled to CRISPR-Cas12a-mediated detection showed a limit of detection of 102 ge/µL in a turnaround time of 2 h. The clinical validation indicated an overall sensitivity of 80%-88%, while for middle and high viral load samples (Cq ≤ 31) the sensitivity was 92%-100%. The specificity was 96%-100% regardless of viral load. Furthermore, we show that the toolkit can be used with the mobile laboratory Bento Lab, potentially enabling LMICs to implement detection services in unattended remote regions.

UnCovid: A versatile, low-cost, and open-source protocol for SARS-CoV-2 RNA detection.

Here, we describe a detailed step-by-step protocol to detect SARS-CoV-2 RNA using RT-PCR-mediated amplification and CRISPR/Cas-based visualization. The optimized assay uses basic molecular biology equipment such as conventional thermocyclers and transilluminators for qualitative detection. Alternatively, a fluorescence plate reader can be used for quantitative measurements. The protocol detects two regions of the SARS-CoV-2 genome in addition to the human RNaseP sample control. Aiming to reach remote regions, this work was developed to use the portable molecular workstation from BentoLab. For complete details on the use and execution of this protocol, please refer to Alcántara et al., 2021.

A low-cost and open-source protocol to produce key enzymes for molecular detection assays.

Here, we describe a detailed step-by-step protocol for the expression, purification, quantification, and activity determination of key enzymes for molecular detection of pathogens. Based on previous reports, we optimized the protocol for LbCas12a, Taq DNA polymerase, M-MLV reverse transcriptase, and TEV protease to make it compatible with minimal laboratory equipment, broadly available in low- and middle-income countries. The enzymes produced with this protocol have been successfully used for molecular detection applications. For complete details on the use and execution of this protocol, please refer to Alcántara et al. (2021a, 2021b).

Circulating miRNA-23b and miRNA-143 Are Potential Biomarkers for In-Stent Restenosis.

In-stent restenosis (ISR) is one of the main complications in patients undergoing percutaneous coronary angioplasty, and microRNAs participate in the contractile-to-synthetic phenotypic switch of vascular smooth muscle cells, a hallmark of restenosis development. MicroRNAs (miRNAs) can be released into circulation from injured tissues, enticing a potential role as noninvasive biomarkers. We aimed to evaluate circulating levels of miRNA-23b, miRNA-143, and miRNA-145 as diagnostic markers of ISR. 142 patients with coronary artery disease undergoing successful angioplasty and a follow-up angiography were included. Subjects were classified according to the degree of obstruction at the angioplasty site into cases (≥50%) or controls (<50%). Total RNA was isolated from plasma to quantify circulating miRNAs levels, and the ROC curves were constructed. Among circulating miRNAs assessed, miRNA-23b and miRNA-143 were significantly lower in cases (miRNA-23b: 18.4x10-5 and miRNA-143: 13.7x10-5) than controls (miRNA-23b: 5.2x10-5, p < 0.0001; miRNA-143: 4.0x10-5, p < 0.0001). Plasma levels of miRNA-145 showed no significant differences. The analysis of the ROC curves showed an area under the curve for miRNA-23b of 0.71 (95% CI: 0.62-0.80, p < 0.0001) and 0.69 for miRNA-143 (95% CI: 0.60-0.78; p < 0.0001). Our data suggest that plasma levels of miRNA-23b and miRNA-143 could be useful as noninvasive biomarkers of ISR.

Synthesis and photophysical properties of novel pyrene-metalloporphyrin dendritic systems.

A novel series of dendronized porphyrins bearing pyrene units in the periphery (Porph-O-Gn) and their metal complexes (M-[Porph-O-Gn]) are reported. The pyrene-containing Frechet-type dendrons up to the first generation were synthesized and further reacted with 5-phenol-10,15,20-triphenylporphyrin via an esterification reaction to afford the desired pyrene-labeled dendronized porphyrins. Later, these compounds were used as ligands to produce the corresponding complexes of Zn2+, Cu2+, Mg2+ and Mn3+. With the compounds in hand, the optical and photophysical properties of the dendritic metalloporphyrins were studied by absorption and fluorescence spectroscopy. The quantum yields, Förster radius and efficiency of energy transfer were determined and discussed as a function of the structure and the donor-acceptor distances, finding an efficient energy transfer from the pyrene moiety to the metallated porphyrin core in each case.

Novel hypertonic saline-sodium hydroxide (HS-SH) method for decontamination and concentration of sputum samples for Mycobacterium tuberculosis microscopy and culture.

This study evaluated a new decontamination and concentration (DC) method for sputum microscopy and culture. Sputum samples from patients with suspected pulmonary tuberculosis (TB) (n=106) were tested using the proposed hypertonic saline-sodium hydroxide (HS-SH) DC method, the recommended N-acetyl-L-cysteine-sodium citrate-sodium hydroxide (NALC-NaOH) DC method and unconcentrated direct smear (Ziehl-Neelsen) techniques for the presence of mycobacteria using Löwenstein-Jensen culture and light microscopy. Of 94 valid specimens, 21 (22.3%) were positive in culture and were further characterized as Mycobacterium tuberculosis. The sensitivity for acid-fast bacilli (AFB) smears was increased from 28.6% using the direct method to 71.4% (HS-SH) and 66.7% (NALC-NaOH) using DC methods. Both concentration techniques were highly comparable for culture (kappa=0.794) and smear (kappa=0.631) for AFB. Thus the proposed HS-SH DC method improved the sensitivity of AFB microscopy compared with a routine unconcentrated direct smear; its performance was comparable to that of the NALC-NaOH DC method for AFB smears and culture, but it was methodologically simpler and less expensive, making it a promising candidate for evaluation by national TB control programmes in developing countries.

[Tobacco smoking in urban and rural adolescents and its association with individual and familiar characteristics]. / Tabaquismo en adolescentes del medio urbano y rural. Caracteristicas individuales y familiares asociadas.

OBJECTIVE: The purpose was to estimate the prevalence of smoking in adolescents and its association with some individual and family characteristics in urban and rural areas. METHODS: An analytic cross-sectional study was carried out in 3161 junior high school students, of eight public and two private urban schools and three public rural schools. Some student behaviors on tobacco use, patterns of smoking and alcohol consumption in the family and socioeconomic status were measured. RESULTS: The prevalence of smoking increased with age. In private urban schools, women had a higher prevalence that males (27.6 % vs. 19.3 %). In rural area schools was higher in males (9.9 % vs. 3.5 %). Individual characteristics associated with smoking were: the curiosity to smoke (OR = 9.7, CI 95 % = 5.8-16.3), having experienced with smoking (OR = 26, CI 95% = 12-56.6) and alcohol consumption (OR = 2.9, CI 95 % = 2.1-3.9). The family characteristics that influence adolescent smoking were the presence of smoking habit in the father (OR = 1.5, CI 95% = 1.1-2) or in the brother or sister (OR = 2, CI 95 % = 1.3-3). The parents living together (OR = 0.6, CI 95 % = 0.4-0.9) was a protective factor. CONCLUSIONS: The exploratory behavior of the adolescent and the patterns of tobacco use and alcohol consumption in the family influenced the adoption of this habit.

Análisis del fenómeno de niños, niñas y adolescentes en situación de trabajo en la ciudad de Villavicencio

Resumen El fenómeno de los niños, niñas y adolescentes en situación de trabajo en Colombia es foco de atención de instituciones gubernamentales y no gubernamentales por lo que es importante generar estrategias que mitiguen su proliferación ya que se afectan derechos fundamentales. Esta situación es objeto de investigación en la ciudad de Villavicencio en el Departamento del Meta; por lo que a través de las verbalizaciones de dos grupos de interés que para el caso fueron los niños, niñas y adolescentes (n = 24) que se encontraban desarrollando actividades de índole laboral por un lado, y por otro, los integrantes del Comité de Erradicación del Trabajo Infantil CETI de dicha ciudad (n = 24). Como resultado de la investigación se observa que hay similitudes y diferencias en las verbalizaciones respecto al fenómeno y como se presenta, ya que se observan discrepancias significativas entre lo que las instituciones dedicadas a erradicar el fenómeno y las actividades que realizan los menores.

Abstract The phenomenon of children and adolescents in work situations in Colombia is a focus of attention of governmental and no governmental institutions, so this is an important issue to generate strategies that mitigate its proliferation since Fundamental Rights are affected, this situation is the subject of research in the city of Villavicencio in the Department of Meta; so through the verbalizations of two interest groups that for the case were the children and adolescents who were actually developing activities of a labor nature and on the other hand, the members of the Committee for the Eradication of Child Labor CETI; therefore, as results of the investigation, it is observed that there are similarities and differences in the verbalizations regarding the phenomenon and how it is presented.

Polimorfismo rs17817449 del gen FTO y su influencia en variables antropométricas de jóvenes chilenos / rs17817449 polymorphism of the fto gene and its influence on anthropometric variables of young Chileans

Determinar la distribución genotípica y la frecuencia alélica del polimorfismo rs17817449 del gen FTO en jóvenes chilenos y su influencia en variables antropométricas. Los 96 sujetos jóvenes (18-25 años), 43 hombres y 53 mujeres fueron evaluados utilizando genotipificación del polimorfismo rs17817449 del gen FTO en TT, TG y GG mediante polimerase chain reaction (PCR), además de una evaluación Kinenatropométrica para determinar las variables asociadas a composición corporal. Las variables fueron analizadas estadísticamente según su distribución paramétrica y el nivel de significancia estadística fue p<0,05. La distribución genotípica del polimorfismo rs17817449 de FTO en jóvenes chilenos fue: TT: 50 %; TG: 42,7 %; GG 7,3 % y la distribución alélica fue: T: 0,7105 y G: 0,2895. No se encontraron diferencias estadísticamente significativas en las variables antropométricas al analizar los participantes según modelo de dominancia del alelo G. Se determinó la distribución genotípica y la frecuencia alélica del polimorfismo rs17817449 del gen FTO en jóvenes chilenos, datos desconocidos hasta este momento. De acuerdo a nuestros resultados, no existen diferencias antropométricas entre personas con diferentes genotipos del polimorfismo rs17817449 de FTO, agrupadas según modelo de dominancia del alelo G.

The rs17817449 polymorphism of the FTO gene in young Chileans and their influence on anthropometric variables. 96 young subjects (18-25 years old), 43 men and 53 women were evaluated using genotyping of the rs17817449 polymorphism of the FTO gene in TT, TG and GG by means of polymerase chain reaction (PCR), in addition to a Kinenatropometric evaluation to determine the variables associated with body composition. The variables were analyzed statistically according to their parametric distribution and the level of statistical significance was p<0.05. The genotypic distribution of the FTO polymorphism rs17817449 in young Chileans was: TT: 50 %; TG: 42.7 %; GG 7.3 % and the allelic distribution was: T: 0.7105 and G: 0.2895. No statistically significant differences were found in the anthropometric variables when analyzing the participants according to model of dominance of the G allele. The genotypic distribution and the allelic frequency of the rs17817449 polymorphism of the gene were determined FTO in Chilean population, data unknown until now. According to our results, there are no anthropometric differences between people with different genotypes of the FTO polymorphism rs17817449, nor according to the dominance model of the G.

Simplifying Recombinant Protein Production: Combining Golden Gate Cloning with a Standardized Protein Purification Scheme.

Recombinant protein production is pivotal in molecular biology, enabling profound insights into cellular processes through biophysical, biochemical, and structural analyses of the purified samples. The demand for substantial biomolecule quantities often presents challenges, particularly for eukaryotic proteins. Escherichia coli expression systems have evolved to address these issues, offering advanced features such as solubility tags, posttranslational modification capabilities, and modular plasmid libraries. Nevertheless, existing tools are often complex, which limits their accessibility and necessitate streamlined systems for rapid screening under standardized conditions. Based on the Golden Gate cloning method, we have developed a simple "one-pot" approach for the generation of expression constructs using strategically chosen protein purification tags like hexahistidine, SUMO, MBP, GST, and GB1 to enhance solubility and expression. The system allows visual candidate screening through mScarlet fluorescence and solubility tags are removable via TEV protease cleavage. We provide a comprehensive protocol encompassing oligonucleotide design, cloning, expression, His-tag affinity chromatography, and size-exclusion chromatography. This method, therefore, streamlines prokaryotic and eukaryotic protein production, rendering it accessible to standard molecular biology laboratories with basic protein biochemical equipment.

Manejo de la paciente diabética durante el climaterio / Therapy of diabetic patient during climateric

En el presente artículo se hace una revisión sobre el manejo de la paciente diabética durante el climaterio. La diabetes en EEUU es la séptima causa de mortalidad, la sexta causa de enfermedad, la principal causa de ceguera y la causa más frecuente de infartos en menores de 30 años. En Colombia se ha calculado que existe un 7 por ciento de diabéticos. En general se puede decir que la diabetis tipo 1 es inmunológica con alteración en la producción de insulina, mientras que la tipo 2, con resistencia a la insulina, se acompaña de obesidad. Las pacientes diabéticas tienen una enfermedad vascular, por lo cual se benefician de los efectos de la terapia hormonal de sustitución sobre prevención de enfermedad coronaria. En la evaluación de ñla paciente diabética es importante tener en cuenta el riesgo aumentado que tiene de sufrir hiperplasia o carcinoma de endometrio. En la mujer con diabetes tipo 1 se debe descartar el compromiso de los órganos blancos, se recomienda dentro de la evaluación inicial realizar dessiometría y prueba de esfuerzo. Al administrar la suplencia se preferirá lavía transdérmica en caso de obesidad o trigllicéridos elevados.

Tratamiento de la vaginitis: Empleo de Tinidazol oral y Tioconazol unguento al 6.5 / Treatment of vaginitis: Use of oral Tinidazol and topic Thioonazole 6.5

Veintisiete pacientes en quienes por sus signos y sintomas se hizo el diagnostico de vaginitis, fueron tratadas con dosis unicas, simultaneas de 2 g orales de tinidazol y una aplicacion de unguento vaginal de tioconazol al 6.5%. El diagnostico clinico fue corroborado por el laboratorio en solo el 41% de los casos. El patogeno mas fecuentemente encontrado fue Candida albicans , en cuatro de estos asociado a Trichomonas vaginalis. Este ultimo se observo en un total de 13 casos (48%), hallandose como patogeno unico en seis casos. La Gardnerella vaginalis se evidencio en cuatro casos, en tres de los cuales asociada a Trichomonas vaginalis. En ningun caso se presento la asociacion de los tres organismos. El tratamiento empleado, por su eficacia, facil dosificacion y buena tolerancia, es una excelente alternativa cuando, por cualquier razon, no pueda hacerse el diagnostico etiologico previo. Para disminuir aun mas la posibiliad de fracasos terapeuticos, se recomienda insistir en el tratamiento al conyuge con tinidazol en todos los casos