In Vitro Analysis of SARS-CoV-2 Spike Protein and Ivermectin Interaction.

The spike (S) protein of SARS-CoV-2 is a molecular target of great interest for developing drug therapies against COVID-19 because S is responsible for the interaction of the virus with the host cell receptor. Currently, there is no outpatient safety treatment for COVID-19 disease. Furthermore, we consider it of worthy importance to evaluate experimentally the possible interaction of drugs (approved by the Food and Drug Administration) and the S, considering some previously in silico and clinical use. Then, the objective of this study was to demonstrate the in vitro interaction of ivermectin with S. The equilibrium dialysis technique with UV-Vis was performed to obtain the affinity and dissociation constants. In addition, the Drug Affinity Responsive Target Stability (DARTS) technique was used to demonstrate the in vitro interaction of S with ivermectin. The results indicate the interaction between ivermectin and the S with an association and dissociation constant of Ka = 1.22 µM-1 and Kd = 0.81 µM, respectively. The interaction was demonstrated in ratios of 1:50 pmol and 1:100 pmol (S: ivermectin) by the DARTS technique. The results obtained with these two different techniques demonstrate an interaction between S and ivermectin previously explored in silico, suggesting its clinical uses to stop the viral spread among susceptible human hosts.

Evaluation of the survival of homofermentative Lacticaseibacillus casei subsp. casei in fermented milk matrix enriched with non-digestible natural fibers.

The physicochemical changes induced by seven different dietary fibers (oat, bamboo, pea, inulin, apple, potato, and wheat) during storage and their effects on the survival of homofermentative Lacticaseibacillus casei subsp. casei (L. casei) in fermented milk matrix were analyzed. For this, an experimental study of the effect of storage time on the microorganisms count and physicochemical properties (pH, titratable acidity, syneresis and viscosity) of milk fermented with L. casei was carried out every two weeks during a storage period of 42 days. Throughout the period studied at 4 °C, no significant differences were found in terms of viscosity values, syneresis rates and L. casei counts, despite the substantial decrease in the pH values. Notably, the substantial increase in the concentration of free hydronium ions (active acidity) in the fiber-enriched matrices during the follow-up period was positively correlated with the L.casei survival. The microbial count determined in all samples was higher than 1 × 107 CFU/g, the minimum value recommended by world organizations for nutraceutical fermented foods. Consequently, the studied prebiotic fibers could be considered in the production of new fermented dairy products with functional properties.

Effect of Long-Term Intake of Nutritive and Non-Nutritive Sweeteners on Metabolic Health and Cognition in Adult Male Rats.

This study evaluated the effects of long-term intake of nutritive sweeteners (NSs) and non-nutritive sweeteners (NNSs) on body weight, food and energy intake, blood pressure, metabolic parameters, and memory retention in rats. Sixty male Sprague-Dawley rats were randomly divided into six groups (n = 10 per group): control (water),10% sucrose (SUC), aspartame (ASP), sucralose (SCA), stevia (STV), and 5% xylitol (XYL). Pure NSs (SUC and XYL) and NNSs were added to the drinking water for 18 weeks. ASP, SCA, and STV dosage was based on the estimated daily intake limit: 4.1, 2.0, and 3.4 mg/kg/day, respectively. Chronic access to NNSs did not result in any difference in total weight gain of the rats, while it was significantly elevated in the SUC group compared with the control and NNSs groups. Food intake was significantly lower in all NNSs groups compared with SUC and control groups. Sweetened beverage intake volumes were significantly diminished in all NNSs groups compared with intake in SUC and control groups. Total calories consumed were lower for the STV and XYL groups compared with all other groups. Blood pressure and glucose metabolism did not differ significantly between the groups. All sweeteners increased total cholesterol, low-density lipoprotein, and triglyceride levels. Short-term memory was significantly impaired in the ASP group in the novel object recognition task, while long-term memory was impaired in SUC and STV groups. These metabolic and behavioral results suggest that the long-term intake of NSs or NNSs can be associated with peripheral and central effects.

Comparative analysis of anthropometric indicators for diagnosing obesity and predicting cardiometabolic risk in Mexican adolescents.

Introduction: Introduction: obesity is a cardiometabolic risk (CMR) factor in adolescents. It represents a public health problem that requires simple diagnostic strategies based on the determination of anthropometric indicators (AIs) such as body mass index (BMI), waist circumference (WC), conicity index (CONI), waist-to-height ratio (WHtR), and body fat percentage (BFP). The purpose of the current contribution was to analyze the utility of the aforementioned AIs for evaluating obesity as a CMR factor in Mexican adolescents. Materials and methods: the design of the study is descriptive and cross-sectional. Nutritional status was established by using various AIs and as a CMR factor under the criteria of WHtR. Participating in the present investigation were 2,000 adolescents from 15 to 17 years of age, of which 1,079 (53.9 %) were female and 921 (46.1 %) were male. All were high school students in Tuxtla Gutiérrez, Chiapas, Mexico. Results and conclusion: the prevalence of obesity was influenced by gender. With the WHtR and BFP, male students exhibited a greater prevalence of obesity, while with the BMI and CONI, the percentage of obesity was similar between the two genders. BMI was significantly correlated with BFP, WC and WHtR, and was one of the AIs with the greatest area under the curve, showing a good capacity to predict cardiometabolic risk. BMI is a simple and adequate tool for diagnosing obesity and predicting CMR in adolescents.

Introducción: Introducción: la obesidad es un factor de riesgo cardiometabolico (RCM) en los adolescentes y representa un problema de salud pública que requiere estrategias diagnosticas sencillas, basadas en la determinación de indicadores antropométricos (IA) como el índice de masa corporal (IMC), la circunferencia de la cintura (CC), el índice de conicidad (ICONI), el índice de cintura-talla (ICT) y el porcentaje de grasa corporal (PGC). El propósito del presente estudio fue analizar la utilidad de diversos IA (IMC, CC, ICONI, ICT y PGC) para evaluar la obesidad como factor de RCM en adolescentes mexicanos. Materiales y métodos: el diseño del estudio fue descriptivo y transversal. Se analizó el estado nutricional mediante varios IA y el factor de RCM bajo los criterios del ICT. Este estudio fue conducido en 2000 adolescentes de entre 15 y 17 años, de los cuales 1079 (53,9 %) eran mujeres y 921 (46,1 %) varones, todos ellos estudiantes de nivel medio superior de Tuxtla Gutiérrez, Chiapas, México. Resultados y conclusión: la prevalencia de la obesidad estuvo influenciada por el género: los varones presentaron una mayor prevalencia de obesidad con ICT y PGC, mientras que con IMC e ICONI, el porcentaje de obesidad fue similar entre ambos géneros. El IMC se correlacionó significativamente con el PGC, la CC y el ICT, y fue uno de los IA con mayor área bajo la curva, mostrando una buena capacidad para predecir el RCM. El IMC representa una herramienta sencilla y adecuada para el diagnóstico de obesidad y predecir el RCM en adolescentes.

Restorative potential of (-)-epicatechin in a rat model of Gulf War illness muscle atrophy and fatigue.

We examined in a rat model of Gulf War illness (GWI), the potential of (-)-epicatechin (Epi) to reverse skeletal muscle (SkM) atrophy and dysfunction, decrease mediators of inflammation and normalize metabolic perturbations. Male Wistar rats (n = 15) were provided orally with pyridostigmine bromide (PB) 1.3 mg/kg/day, permethrin (PM) 0.13 mg/kg/day (skin), DEET 40 mg/kg/day (skin) and were physically restrained for 5 min/day for 3 weeks. A one-week period ensued to fully develop the GWI-like profile followed by 2 weeks of either Epi treatment at 1 mg/kg/day by gavage (n = 8) or water (n = 7) for controls. A normal, control group (n = 15) was given vehicle and not restrained. At 6 weeks, animals were subjected to treadmill and limb strength testing followed by euthanasia. SkM and blood sampling was used for histological, biochemical and plasma pro-inflammatory cytokine and metabolomics assessments. GWI animals developed an intoxication profile characterized SkM atrophy and loss of function accompanied by increases in modulators of muscle atrophy, degradation markers and plasma pro-inflammatory cytokine levels. Treatment of GWI animals with Epi yielded either a significant partial or full normalization of the above stated indicators relative to normal controls. Plasma metabolomics revealed that metabolites linked to inflammation and SkM waste pathways were dysregulated in the GWI group whereas Epi, attenuated such changes. In conclusion, in a rat model of GWI, Epi partially reverses detrimental changes in SkM structure including modulators of atrophy, inflammation and select plasma metabolites yielding improved function.

Association of anthropometric indicators to evaluate nutritional status and cardiometabolic risk in Mexican teenagers / Asociación de indicadores antropométricos para evaluar el estado nutricional y el riesgo cardiometabólico en adolescentes mexicanos

Introduction: anthropometric indicators (AIs) such as waist circumference (WC), body mass index (BMI), waist/hip index (WHpI), waist/height index (WHtI) and body fat percentage (BFP) are useful tools for the diagnosis of nutritional status (NS) in adolescents. Each of these parameters has advantages and disadvantages. The purpose of the present study was to analyze the association of these AIs (WC, BMI, WHpI, WHtI, and BFP) to evaluate nutritional status and estimate the cardiometabolic risk (CMR) in Mexican adolescents. Material and method: in a cross-sectional descriptive study, the NS was analyzed through various AIs and CMR with the WHtI criteria. Nine hundred and seventeen adolescents between 15 and 17 years old participated in the study, of whom 488 (52.9%) were female and 429 (47.1%) male, all students of middle school in Tuxtla Gutiérrez, Chiapas, Mexico. Results and conclusion: women presented a higher prevalence of obesity according to most indicators. The WHtI was the parameter that detected the highest prevalence of obesity (31%), correlating with the BMI and the BFP. Moreover, there was evidence of a significant relation between NS (assessed by all the anthropometric indicators) and CMR. The WHtI could be considered as an adequate tool for the diagnosis of obesity associated with CMR in adolescents

Introducción: los indicadores antropométricos (IA) como la circunferencia de cintura (CC), el índice de masa corporal (IMC), el índice cintura/ cadera (ICC), el índice cintura/talla (ICT) y el porcentaje de grasa corporal (PGC) son herramientas útiles para el diagnóstico del estado nutricional (EN) en los adolescentes. Sin embargo, cada uno de estos IA presentan ventajas y desventajas. El propósito del presente estudio fue analizar la asociación de los IA (IMC, CC, ICC, ICT y PGC) para evaluar el EN y estimar el riesgo cardiometabólico (RCM) en adolescentes mexicanos. Material y método: el diseño del estudio fue descriptivo transversal. Se analizó el EN a través de diversos IA y el RCM bajo los criterios de ICT. Este estudio fue conducido en 917 adolescentes de entre 15 y 17 años, de los cuales 488 (52,9%) eran mujeres y 429 (47,1%) varones, todos estudiantes de nivel medio superior de Tuxtla Gutiérrez, Chiapas, México. Resultados y conclusión: las mujeres presentaron mayor prevalencia de obesidad con la mayoría de los IA utilizados. El ICT fue el IA que detectó mayor prevalencia de obesidad (31%), correlacionándose con el IMC y el PGC. Además, se evidenció una asociación significativa entre el EN valorado por todos los IA y el RCM. El ICT podría ser considerado como una herramienta adecuada para el diagnóstico de obesidad asociada a RCM en adolescentes

Association of anthropometric indicators to evaluate nutritional status and cardiometabolic risk in Mexican teenagers.

INTRODUCTION: Introduction: anthropometric indicators (AIs) such as waist circumference (WC), body mass index (BMI), waist/hip index (WHpI), waist/height index (WHtI) and body fat percentage (BFP) are useful tools for the diagnosis of nutritional status (NS) in adolescents. Each of these parameters has advantages and disadvantages. The purpose of the present study was to analyze the association of these AIs (WC, BMI, WHpI, WHtI, and BFP) to evaluate nutritional status and estimate the cardiometabolic risk (CMR) in Mexican adolescents. Material and method: in a cross-sectional descriptive study, the NS was analyzed through various AIs and CMR with the WHtI criteria. Nine hundred and seventeen adolescents between 15 and 17 years old participated in the study, of whom 488 (52.9%) were female and 429 (47.1%) male, all students of middle school in Tuxtla Gutiérrez, Chiapas, Mexico. Results and conclusion: women presented a higher prevalence of obesity according to most indicators. The WHtI was the parameter that detected the highest prevalence of obesity (31%), correlating with the BMI and the BFP. Moreover, there was evidence of a significant relation between NS (assessed by all the anthropometric indicators) and CMR. The WHtI could be considered as an adequate tool for the diagnosis of obesity associated with CMR in adolescents.

INTRODUCCIÓN: Introducción: los indicadores antropométricos (IA) como la circunferencia de cintura (CC), el índice de masa corporal (IMC), el índice cintura/cadera (ICC), el índice cintura/talla (ICT) y el porcentaje de grasa corporal (PGC) son herramientas útiles para el diagnóstico del estado nutricional (EN) en los adolescentes. Sin embargo, cada uno de estos IA presentan ventajas y desventajas. El propósito del presente estudio fue analizar la asociación de los IA (IMC, CC, ICC, ICT y PGC) para evaluar el EN y estimar el riesgo cardiometabólico (RCM) en adolescentes mexicanos. Material y método: el diseño del estudio fue descriptivo transversal. Se analizó el EN a través de diversos IA y el RCM bajo los criterios de ICT. Este estudio fue conducido en 917 adolescentes de entre 15 y 17 años, de los cuales 488 (52,9%) eran mujeres y 429 (47,1%) varones, todos estudiantes de nivel medio superior de Tuxtla Gutiérrez, Chiapas, México. Resultados y conclusión: las mujeres presentaron mayor prevalencia de obesidad con la mayoría de los IA utilizados. El ICT fue el IA que detectó mayor prevalencia de obesidad (31%), correlacionándose con el IMC y el PGC. Además, se evidenció una asociación significativa entre el EN valorado por todos los IA y el RCM. El ICT podría ser considerado como una herramienta adecuada para el diagnóstico de obesidad asociada a RCM en adolescentes.

Choline acetyltransferase and TrkA expression, as well as the improvement in cognition produced by E2 and P4 in ovariectomized rats, are blocked by ICI 182 780 and RU486.

Treatment with 17-ß estradiol and progesterone improves the performance of ovariectomized rats in an autoshaping learning task, representing cognitive improvement. To test whether this is attributable to genomic mechanisms, the antiestrogen ICI 182 780 or antiprogesterone RU486 was injected into ovariectomized animals primed previously with estrogen or progesterone, respectively. Compared with the vehicle control, each hormone administered alone produced an elevated expression of choline acetyltransferase and TrkA, along with an improvement in performance on the behavioral test. E2+ICI reverted the increase in these two proteins. However, RU alone elicited higher ChAT expression. With this exception, there was a clear linear regression between the number of conditioned responses and the level of ChAT and TrkA in the basal forebrain. The results suggest that TrkA may be more important than ChAT for regulating autoshaping learning tasks, and that genomic mechanisms in the basal forebrain could possibly underlie hormonal improvement of cognition.

Agonist E-6837 and antagonist SB-271046 of 5-HT6 receptors both reverse the depressive-like effect induced in mice by subchronic ketamine administration.

Major depression is one of the most common affective disorders caused by schizophrenia. The administration of N-methyl-D-aspartate receptor antagonists, such as ketamine, can reproduce the negative and affective symptoms of this disorder in animals. Preclinical studies have shown that 5-HT6 receptor (5-HT6R) agonists and antagonists have a considerable antipsychotic response. The aim of the present study was to evaluate the effect of an acute treatment with an agonist, E-6837, and an antagonist, SB-271046, of 5-HT6R on the immobility induced in mice by a subchronic ketamine regimen (5 days; 10 mg/kg/day, intraperitoneal). Repeated ketamine administration alone increased the immobility time in the forced-swimming test and the tail-suspension test. E-6837 at 10 and 20 mg/kg caused a significant reduction of immobility in the tail-suspension test and forced-swimming test, respectively. Interestingly, SB-271046 (10 mg/kg) also elicited an antidepressant-like effect in both tests. The current findings suggest an important role for these 5-HT6R ligands as mood modulators. However, it is necessary to explore the physiological mechanisms involved in this process in greater detail.

Tibolone modulates neuronal plasticity through regulating Tau, GSK3ß/Akt/PI3K pathway and CDK5 p35/p25 complexes in the hippocampus of aged male mice.

Aging is a key risk factor for cognitive decline and age-related neurodegenerative disorders. Also, an age-related decrease in sex steroid hormones may have a negative impact on the formation of neurofibrillary tangles (NFTs); these hormones can regulate Tau phosphorylation and the principal kinase GSK3ß involved in this process. Hormone replacement therapy decreases NFTs, but it increases the risk of some types of cancer. However, other synthetic hormones such as tibolone (TIB) have been used for hormone replacement therapy. The aim of this work was to evaluate the long-term effects of TIB (0.01 mg/kg and 1 mg/kg, intragastrically for 12 weeks) on the content of total and hyperphosphorylated Tau (PHF-1) proteins and the regulation of GSK3ß/Akt/PI3K pathway and CDK5/p35/p25 complexes in the hippocampus of aged male mice. We observed that the content of PHF-1 decreased with TIB administration. In contrast, no changes were observed in the active form of GSK3ß or PI3K. TIB decreased the expression of the total and phosphorylated form of Akt while increased that of p110 and p85. The content of CDK5 was differentially modified with TIB: it was increased at low doses and decreased at high doses. When we analyzed the content of CDK5 activators, an increase was found on p35; however, the content of p25 decreased with administration of low dose of TIB. Our results suggest a possible mechanism of action of TIB in the hippocampus of aged male mice. Through the regulation of Tau and GSK3ß/Akt/PI3K pathway, and CDK5/p35/p25 complexes, TIB may modulate neuronal plasticity and regulate learning and memory processes.

Chronic administration of tibolone modulates anxiety-like behavior and enhances cognitive performance in ovariectomized rats.

Hormone replacement therapy (HRT) may be prescribed to prevent the symptoms of menopause. This therapy may include estrogenic and/or progestin components and may increase the incidence of endometrial and breast cancers. Tibolone (TIB), which is also made up of estrogen and progestin components, is often used to reduce the impact of HRT. However, the effect of TIB on the processes of learning, memory and anxiety has yet to be fully elucidated. The aim of this study was to evaluate the long-term effect on learning, memory processes and anxiety in ovariectomized rats caused by different doses of TIB (0 mg/kg, 0.01 mg/kg, 0.1 mg/kg 1.0 mg/kg and 10 mg/kg, administered daily via the oral route for 18 weeks). Two behavioral animal models, the autoshaping and T maze models were employed. The concentrations of acetyl choline transferase (ChAT) and tryptophan hydroxylase (TPH) in the hippocampus were directly measured by Western blot. No significant changes were observed in the autoshaping model and spontaneous activity test. In the T maze, increased latency was observed with TIB doses of 1 and 10 mg/kg compared to the vehicle. We observed that the ChAT content decreased with increasing doses of TIB, whereas TPH content increased with doses of 1 and 10 mg/kg of TIB. These data indicate that high doses of TIB improved emotional learning, which may be related to the modulation of the cholinergic and serotonergic systems by TIB.

Short- and long-term treatment with estradiol or progesterone modifies the expression of GFAP, MAP2 and Tau in prefrontal cortex and hippocampus.

AIMS: We analyzed the effects of the short- and long-term administration of estradiol (E2) or progesterone (P4) after ovariectomy on the expression of MAP2, Tau and GFAP in prefrontal cortex and hippocampus. MAIN METHODS: Sprague Dawley rats were ovariectomized and immediately treated with E2 or P4 for 2 or 18 weeks. At the end of treatments, hippocampus and prefrontal cortex were excised, proteins were extracted and MAP2, Tau and GFAP were analyzed by Western blot. KEY FINDINGS: MAP2 and Tau content was not modified by E2 in the prefrontal cortex. On the contrary, P4 decreased MAP2 content after a short-term treatment, while it increased that of MAP2 and TAU in this brain region after a long-term treatment. E2 increased MAP2 content in hippocampus. In this region, short-term administration of P4 increased that of MAP2. GFAP content was diminished after a long-term administration of P4 in hippocampus. SIGNIFICANCE: Current data emphasize the importance of short- and long-term sex steroid treatment on neuronal and glial cytoskeletal proteins expression.

Effects of short-term hormonal replacement on learning and on basal forebrain ChAT and TrkA content in ovariectomized rats.

It has been proposed that sex steroid hormones improve performance in some cognitive tasks by regulating the basal forebrain cholinergic function. However, the molecular basis of such influence still remains unknown. Current study analyzed the performance of ovariectomized rats in an autoshaping learning task after a short-term treatment with 17ß-estradiol (E2: 4 and 40µg/kg) and/or progesterone (P4: 4mg/kg). These results were correlated with basal forebrain choline acetyltransferase (ChAT) and TrkA protein content. The high dose of E2 enhanced both acquisition in the autoshaping task and the content of ChAT and TrkA. P4 treatment increased ChAT and TrkA content without affecting performance of rats in the autoshaping learning task. Interestingly, the continuous and simultaneous administration of E2 plus P4 did not significantly modify behavioral and biochemical evaluated parameters. These results address the influence of both E2 and P4 on cholinergic and TrkA activity and suggest that the effects of ovarian hormones on cognitive performance involve basal forebrain cholinergic neurons.

Inhibition of acetylcholinesterase by two arylderivatives: 3a-Acetoxy-5H-pyrrolo (1,2-a) (3, 1)benzoxazin- 1,5-(3aH)-dione and cis-N-p-Acetoxy-phenylisomaleimide.

Two arylderivatives, 3a-Acetoxy-5H-pyrrolo(1,2-a) (3,1)benzoxazin-1,5-(3aH)-dione 3 and cis-N-p-Acetoxy-phenylisomaleimide 4, were synthesized from anthranilic acid and para-aminophenol, respectively. The inhibitory effects of these compounds on acetylcholinesterase (AChE) activity were evaluated in vitro as well as by docking simulations. Both compounds showed inhibition of AChE activity (Ki = 4.72 +/- 2.3 microM for 3 and 3.6 +/- 1.8 microM for 4) in in vitro studies. Moreover, they behaved as irreversible inhibitors and made pi-pi interaction with W84 and hydrogen bonded with S200 and Y337 according to experimental data and docking calculations. The docking calculations showed deltaG bind (kcal/mol) of - 9.22 for 3 and - 8.58 for 4. These two compounds that can be use as leads for a new family of anti-Alzheimer disease drugs.

Ex vivo anticholinesterase activity of benzoic acid derivatives.

p-Aminobenzoic acid (p-ABA) derivatives were evaluated as acetylcholinesterase inhibitors (AChEIs). Finding a correlation between AChE activity with the partition coefficient (log P) and the highest occupied molecular orbital (HOMO) energy of the compounds.