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Bioactive compounds in plant-based food have protective effects against metabolic alterations, including non-alcoholic fatty liver disease (NAFLD). Bean leaves are widely cultivated in the world and are a source of dietary fiber and polyphenols. High fat/high fructose diet animal models promote deleterious effects in adipose and non-adipose tissues (lipotoxicity), leading to obesity and its comorbidities. Short-term supplementation of bean leaves exhibited anti-diabetic, anti-hyperlipidemic, and anti-obesity effects in high-fat/high-fructose diet animal models. This study aimed to evaluate the effect of bean leaves supplementation in the prevention of lipotoxicity in NAFLD and contribute to elucidating the possible mechanism involved for a longer period of time. During thirteen weeks, male Wistar rats (n = 9/group) were fed with: (1) S: Rodent Laboratory Chow 5001® (RLC); (2) SBL: 90% RLC+ 10% dry bean leaves; (3) H: high-fat/high-fructose diet; (4) HBL: H+ 10% of dry bean leaves. Overall, a HBL diet enhanced impaired glucose tolerance and ameliorated obesity, risk factors in NAFLD development. Additionally, bean leaves exerted antioxidant (↑serum GSH) and anti-inflammatory (↓mRNA TNFα in the liver) effects, prevented hepatic fat accumulation by enhanced ↑mRNA PPARα (β oxidation), and enhanced lipid peroxidation (↓liver MDA). These findings suggest that bean leaves ameliorated hepatic lipotoxicity derived from the consumption of a deleterious diet.

Time-restricted feeding and food enriched in polyphenols are strategies to prevent or reduce metabolic disorders. Bean leaves (Phaseolus vulgaris L.) are a recognized source of polyphenolic compounds, whose effects on metabolic pathways are not well studied. We evaluated the combined effects of dietary supplementation with Phaseolus vulgaris leaves (10% w/w) (BL) and a 7-h daytime-restricted feeding protocol (RF) under a hypercaloric diet (high fat + high fructose) (HFFD) on the metabolic parameters related to glucose and lipid handling. Adult male Wistar rats were treated for 8 weeks with standard and HFFD diets with or without BL. The results showed that RF improved metabolic alterations induced by HFFD (e.g., hepatic steatosis, increased triacylglycerols, and serum lipoproteins). Supplementation with BL significantly enhanced this effect and downregulated the mRNA expression of carbohydrate and lipid metabolism genes in the liver. These results indicate that dietary supplementation with BL enhances the benefits elicited by RF.

We present the purification and characterization of the two most abundant isoforms of lectins isolated from Tepary bean (Phaseolus acutifolius) seeds, which have been shown to differentially affect the survival of different cancer cells. They were separated by concanavalin A-affinity chromatography. After purification, to release the N-glycans, they were digested with the endoglycosidases PNGase and Glycanase A. Fractions resulted from the hydrolysis products were analyzed to determine their carbohydrate composition. Mass spectrometry data indicated that both isoforms contained high mannose glycans being mannose 6 the most abundant form. Furthermore, based on sequence Ans-X-Ser/Thr, where X is any amino acid except proline, a glycosylation site was determined on asparagine 36. When their metal requirement to preserve their biological activity was determined, the lectins showed differences. While lectin A (LA) agglutination activity was best in the presence of magnesium, lectin B (LB) was best with calcium. Additionally, only LA exhibited affinity to human type-A erythrocytes. Although both lectins showed small differences in their properties, an identical structure-model for both lectins was generated by the homology modelling process. Also, the analysis of ligand binding sites and in silico glycosylation were achieved. Molecular docking with colon adenocarcinoma associated-N-glycans revealed some highly possible interactions and, on the other hand, that N-glycan interaction zones of Tepary bean lectins is not restricted to the carbohydrate binding domain but to an extended part of their surface, which could lead new strategies to explain their biological activity.

High-fat/high-fructose diets promote early metabolic disorders in weight and lipid and glucose metabolism. Bioactive compounds such as polyphenols and fiber present in plant-based food prevent the development of metabolic disorders. The objective of the present study was to evaluate the effect of Phaseolus vulgaris L. Flor de Mayo Eugenia (FME) bean leaves on early metabolic alterations in male Wistar rats fed a high-fat/high-fructose diet. After proximate and chemical analysis of FME bean leaves, thirty-six male Wistar rats (ethical approval 06FCN2019 and 77FCN2019) were randomly assigned to one of four groups: 1) standard diet (S) fed with Rodent Laboratory Chow 5001®; 2) standard diet + 10% dry FME bean leaves (SBL); 3) high-fat (lard) and high-fructose diet (H); and 4) high-fat/high-fructose diet + 10% dry FME bean leaves (HBL). The study was carried out for six weeks. Group H exhibited early metabolic alterations compared to Group S: final weight gain (↑15%), abdominal fat accumulation (waist circumference, ↑11%), triglycerides (↑30%), glucose (↑16%), insulin resistance (HOMA-IR, ↑32%), and fecal triglycerides (↑284%) and decreased total short-chain fatty acids (SCFAs, ↓17%). FME bean leave supplementation (HBL) prevented body weight gain (↓12%), abdominal fat accumulation (waist circumference, ↓10%), and early insulin resistance (glucose area under the curve, ↓6%) compared to Group H. The supplementary bean leave diet increased SCFA production (↑54%), most likely mediated by the fiber and polyphenols present in the leaves. Therefore, bean leaves are a low-cost alternative for human nutritional care and prevention of early metabolic alterations.

Obesity is one of the main public health problems in Mexico and the world and one from which a large number of pathologies derive. Single nucleotide polymorphisms (SNPs) of various genes have been studied and proven to contribute to the development of multiple diseases. SNPs of the leptin pathway have been associated with the control of hunger and energy expenditure as well as with obesity and type 2 diabetes mellitus. Therefore, the present work focused on determining the association between anthropometric markers and biochemical and dietary factors related to obesity and SNPs of leptin pathway genes, such as the leptin gene (LEP), the leptin receptor (LEPR), proopiomelanocortin (POMC), prohormone convertase 1 (PCSK1), and the melanocortin 4 receptor (MC4R). A population of 574 young Mexican adults of both sexes, aged 19 years old on average and without metabolic disorders previously diagnosed, underwent a complete medical and nutritional evaluation, biochemical determination, and DNA extraction from the blood; DNA samples were subsequently genotyped. Association analyses between anthropometric, biochemical, and dietary variables with SNPs were performed using binary logistic regressions (p-value = 0.05). Although the sampled population did not have previously diagnosed diseases, the evaluation results showed that 33% were overweight or obese according to BMI and 64% had non-clinically elevated levels of body fat. From the 74 SNP markers analyzed from the five previously mentioned genes, 62 showed polymorphisms within the sampled population, and only 35 of these had significant associations with clinical variables. The risk associations (OR > 1) occurred between clinical markers with elevated values for waist circumference, waist–height index, BMI, body fat percentage, glucose levels, insulin levels, HOMA-IR, triglyceride levels, cholesterol levels, LDL-c, low HDL-c, carbohydrate intake, and protein intake and SNPs of the LEP, LEPR, PCSK1, and MC4R genes. On the other hand, the protective associations (OR < 1) were associated with markers including elevated values for insulin, HOMA-IR, cholesterol, c-LDL, energy intake > 2440 Kcal/day, and lipid intake and SNPs of the LEP and LEPR genes and POMC. The present study describes associations between SNPs in leptin pathway genes, revealing positive and negative interactions between reported SNPs and the clinical markers related to obesity in a sampled Mexican population. Hence, our results open the door for the further study of new genetic variants and their influence on obesity.

High-fat/high-fructose diets promote early metabolic disorders in weight, lipid and glucose metabolism. Bioactive compounds such as polyphenols and fiber present in plant-based food prevent metabolic disorders development. The objective of the present study was to evaluate the effect of bean leaves on early metabolic alterations in Wistar male rats fed with high-fat/high-fructose diet. After proximate and chemical analysis of bean leaves, thirty-six Wistar male rats (ethical approval 06FCN2019 and 77FCN2019) were randomly assigned into one of four groups: 1) Standard diet (S) fed with Rodent Laboratory Chow 5001®, 2) Standard diet + 10% of dry bean leaves (SBL), 3) high fat (lard) and fructose diet (H) and 4) high-fat/high-fructose diet + 10% of dry bean leaves (HBL). The study was carried out for 6 weeks. Group H exhibited early metabolic alterations compared to S, final weight gain (↑15%), abdominal fat accumulation (↑11% waist circumference), triglycerides (↑30%), glucose (↑16%), insulin resistance (↑HOMA-IR 32%), fecal triglycerides (↑284%) and decreased total short chain fatty acid (↓SFCA, 17%). Bean leaves supplementation (HBL) compared to group H prevented body weight gain (↓12%), abdominal fat accumulation (waist circumference, ↓10%), early insulin resistance (glucose area under the curve, ↓6%). Probably mediated by the fiber and polyphenols present in the leaves, supplementary bean leaves-diet increased SFCA production (↑54%). Therefore, bean leaves are a low-cost alternative for human nutritional care and prevention of early metabolic alterations.

The Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) exhibits differential cytotoxicity on colon cancer cells and inhibition of early tumorigenesis in the colon (50 mg/kg, three times per week, for 6 weeks). TBLF showed low toxicity with the ability to activate the immune system; however, some adverse effects are the loss in body weight gain, intestinal atrophy, and pancreatic hyperplasia. After a recovery period of 2 weeks after treatment, reversion of pancreatic hyperplasia but no recovery of intestinal atrophy was observed. As TBLF has shown anticancer effects on the colon, it is important to characterize the adverse effects and how they can be reversed. Sprague Dawley rats were administered with TBLF (50 mg/kg) for 6 weeks, three times per week, and then allowed to recover for 6 weeks post-treatment. After TBLF administration, small intestine atrophy, villus atrophy, and cryptic hyperplasia were confirmed, as well as increased intestinal mucus production, increased permeability and a decrease in the apparent ileal digestibility of crude proteins. The colon showed damage in the simple prismatic tissue and decreased crypt depth, and changes in microbiota and a decrease in the apparent fecal digestibility of crude protein were determined. Our results show that the adverse effects provoked by TBLF were partially reversed after 6 weeks of recovery post-treatment, suggesting that increasing the recovery period it could be possible to reverse all adverse effects observed.

Hybridization is defined as the interbreeding of individuals from two populations distinguishable by one or more heritable characteristics. Snake hybridization represents an interesting opportunity to analyze variability and how genetics affect the venom components between parents and hybrids. Snake venoms exhibit a high degree of variability related to biological and biogeographical factors. The aim of this work is to analyze the protein patterns and enzymatic activity of some of the main hemotoxic enzymes in snake venoms, such as serine proteases (trypsin-like, chymotrypsin-like, and elastase-like), metalloproteases, hyaluronidases, and phospholipase A

(1) Background: obesity is a global public health problem; various factors have been associated with this disease, and genetic factors play a very important role. Previous studies in multiple populations have associated a gene with fat mass and obesity (FTO). Thus, the present work aims to identify and determine associations between genetic variants of FTO with indicators of overweight and obesity in the Mexican population. (2) Methods: a total of 638 subjects were evaluated to compile data on body mass index (BMI), the percentage of body fat (%BF), the waist circumference (WC), the serum levels of triglycerides (TG), and food consumption. A total of 175 genetic variants in the FTO gene were sampled by a microarray in the evaluated population, followed by association statistical analyses and comparisons of means. (3) Results: a total of 34 genetic variants were associated with any of the 6 indicators of overweight and obesity, but only 15 showed mean differences using the recessive model after the Bonferroni correction. The present study shows a wide evaluation of FTO genetic variants associated with a classic indicator of overweight and obesity, which highlights the importance of genetic analyses in the study of obesity.

Muscle growth is regulated by several factors including the growth differentiation factor 8, known as myostatin, an inhibitor of myocyte differentiation and proliferation. Research on myostatin regulation was already conducted to improve growth rates in farmed animals, including aquatic species. To explore the effects of myostatin inactivation in a commercial marine fish (spotted rose snapper, Lutjanus guttatus) in vivo, we induced post-transcriptional silencing (knockdown) of myostatin-1 (mstn-1) by injecting dsiRNA directly into the muscle of juvenile fish (87 days post-hatch) using a commercial polymer as vehicle. Results show a significant decrease in mstn-1 expression starting at 2 days after injection and for up to 5 days. Knockdown of mstn-1 caused muscle fiber hypertrophy (but not hyperplasia); however, there were no significant changes in weight or length. Although still experimental, this study provides evidence that temporary knockdown of mstn-1 in a commercial marine fish in vivo promotes fiber hypertrophy and therefore could potentially help grow-out programmes in fish aquaculture.

Aberrant crypt foci (ACF) is the precursor lesion of colorectal carcinogenesis (CRC), one of the most common malignancies in the world. Many studies have reported that people with higher p...

Since the start of the COVID-19 outbreak, the race for testing new platforms designed to confer immunity against SARS-CoV-2, has been rampant and unprecedented, leading to conditional emergency authorization of various vaccines. Despite progress on early multidrug therapy for COVID-19 patients, the current mandate is to immunize the world population as quickly as possible. The lack of thorough testing in animals prior to clinical trials, and authorization based on safety data generated during trials that lasted less than 3.5 months, raise questions regarding vaccine safety. The recently identified role of SARS-CoV-2 Spike glycoprotein for inducing endothelial damage characteristic of COVID-19, even in absence of infection, is extremely relevant given that most of the authorized vaccines induce endogenous production of Spike. Given the high rate of occurrence of adverse effects that have been reported to date, as well as the potential for vaccine-driven disease enhancement, Th2-immunopathology, autoimmunity, and immune evasion, there is a need for a better understanding of the benefits and risks of mass vaccination, particularly in groups excluded from clinical trials. Despite calls for caution, the risks of SARS-CoV-2 vaccination have been minimized or ignored by health organizations and government authorities. As for any investigational biomedical program, data safety monitoring boards (DSMB) and event adjudication committees (EAC), should be enacting risk mitigation. If DSMBs and EACs do not do so, we will call for a pause in mass vaccination. If DSMBs and EACs do not exist, then vaccination should be halted immediately, in particular for demographic groups at highest risk of vaccine-associated death or serious adverse effects, during such time as it takes to assemble these boards and commence critical and independent assessments. We urge for pluralistic dialogue in the context of health policies, emphasizing critical questions that require urgent answers, particularly if we wish to avoid a global erosion of public confidence in science and public health.

Background Vitamin D (VD) has been classically associated with calcium homeostasis and bone mineral density since it has a key role on mineralization and resorption. Immunomodulatory effects have been attributable to VD; low concentrations of VD have been associated with elevation of inflammatory markers. Inflammatory autoimmune diseases, such as multiple sclerosis (MS), a chronic neurodegenerative suffering, whose etiology is still unknown, is directly related to an increase in pro-inflammatory cytokines such as interleukin 17 and interleukin 1β who play an important role in this physiopathology. Nowadays, even though additional studies have linked MS's clinical signs with low VD concentration, there is scarce information of this association in people from regions with sufficient sun exposure. The aim of this study was to evaluate serum VD and cytokine concentrations, and bone density, in Mexican people with MS. Methods Vitamin D (25OHD), interleukin 1β, interleukin 6 and interleukin 17 concentrations of twenty-five volunteers with MS were determined by enzyme-linked immunosorbent assay. Bone mineral density and body composition assessment was performed by dual energy X-Ray absorptiometry. Results A mean concentration of 17.3 ± 4.6 ng/ml of 25OHD was obtained, in a range of 5.15 to 25.71 ng/ml; when international advisory bodies thresholds were applied 76% of the participants exhibited some degree of VD inadequacy. Pro-inflammatory markers were detectable among the participants: interleukin 1β in 100%, interleukin 6 in 64%, whereas interleukin 17 was found in 24% of the volunteers. Bone mineral density below the expected for the age was found in 8% of the participants, with lumbar spine as the most affected anatomic region. Non-significant correlations were found between VD and bone mineral density (Z-score) or pro-inflammatory markers. Conclusion Although non-significant correlations were found between VD and bone mineral density or cytokines, it is important to highlight that an important percentage of our participants exhibited some degree of VD inadequacy, an unknown fact for them, since these are not included in routine clinical evaluations. The low concentrations of VD among this sample regardless of annual UVB sun exposure may suggest the involvement of endogenous and not environmental factors. Further works are needed in order to deepen the physiological causes and effects of VD deficiency in people with MS.

The production of heterologous proteins for medical use is an important area of interest. The optimization of the bioprocesses includes the improvement of time, costs, and unit operations. Our study shows that a lectin fraction from Tepary bean (Phaseolus acutifolius) (TBLF) has cytotoxic effects on colon cancer cells and in vivo antitumorigenic activity. However, the low-yield, time-consuming, and expensive process made us focus on the development of a strategy to obtain a recombinant lectin using engineered Pichia pastoris yeast. Pure glycerol is one of the most expensive supplies, therefore, we worked on process optimization using crude glycerol from biodiesel production. Recombinant lectin (rTBL-1) production and purification were evaluated for the first time by an experimental design where crude glycerol (G65) was used and compared against pure glycerol (G99) in a controlled stirred-tank bioreactor with a fed-batch system. The recombinant lectin was detected and identified by SDS-PAGE, Western blot, and UHPLC–ESI–QTOF/MS analysis. The results show that the recombinant lectin can be produced from G65 with no significant differences with respect to G99: the reaction rates were 2.04 and 1.43 mg L−1 h−1, and the yields were 264.95 and 274.67 mgL−1, respectively. The current low cost of crude glycerol and our results show the possibility of producing heterologous proteins using this substrate with high productivity.

Tepary bean (Phaseolus acutifolius) lectins exhibit differential in vitro and in vivo biological effects, but their gastrointestinal interactions and digestion have not yet been assessed. This work aimed to evaluate the changes of a recombinant Tepary bean lectin (rTBL-1) through an in vitro and ex vivo gastrointestinal process. A polyclonal antibody was developed to selectively detect rTBL-1 by Western blot (WB) and immunohistochemical analysis. Everted gut sac viability was confirmed until 60 min, where protein bioaccessibility, apparent permeability coefficient, and efflux ratio showed rTBL-1 partial digestion and absorption. Immunoblot assays suggested rTBL-1 internalization, since the lectin was detected in the digestible fraction. The immunohistochemical assay detected rTBL-1 presence at the apical side of the small intestine, potentially due to the interaction with the intestinal cell membrane. The in silico interactions between rTBL-1 and some saccharides or derivatives showed high binding affinity to sialic acid (−6.70 kcal/mol) and N-acetylglucosamine (−6.10 kcal/mol). The ultra-high-performance liquid chromatography–electron spray ionization–quantitative time-of-flight coupled to mass spectrometry (UHPLC–ESI–QTOF/MS) analysis showed rTBL-1 presence in the gastric content and the non-digestible fraction after intestinal simulation conditions. The results indicated that rTBL-1 partially resisted the digestive conditions and interacted with the intestinal membrane, whereas its digestion allowed the absorption or internalization of the protein or the derivative peptides. Further purification of digestion samples should be conducted to identify intact rTBL-1 protein and digested peptides to assess their physiological effects.

The elongation of very long chain fatty acids (ELOVL) is a family of seven enzymes that have specific functions in the synthesis of fatty acids. Some have been shown to be related to insulin secretion (ELOVL2), and in the lipid profile (ELOVL6) and patients with various pathologies. The present work focused on the study of ELOVL polymorphs with clinical markers of non-communicable chronic diseases in the Mexican population. A sample of 1075 participants was obtained, who underwent clinical, biochemical, and nutritional evaluation, and a genetic evaluation of 91 genetic variants of ELOVL was considered (2&ndash, 7). The results indicate a 33.16% prevalence of obesity by body mass index, 13.84% prevalence of insulin resistance by homeostatic model assessment (HOMA) index, 7.85% prevalence of high cholesterol, and 20.37% prevalence of hypercholesterolemia. The deprived alleles showed that there is no association between them and clinical disease risk markers, and the notable finding of the association studies is that the ELOVL2 variants are exclusive in men and ELVOL7 in women. There is also a strong association of ELOVL6 with various markers. The present study shows, for the first time, the association between the different ELOVLs and clinical markers of chronic non-communicable diseases.

BackgroundThe need for safe and effective antiviral treatments is pressing given the number of viral infections that are prevalent in animal and human populations, often causing devastating economic losses and mortality. Informal accounts of anecdotal use of chlorine dioxide (ClO2), a well-known disinfectant and antiseptic, in COVID-19 patients has raised concern about potential toxicity, but also raises the question that ClO2 might elicit antiviral effects, a possibility that has never been examined in vivo in any animal model. Here, we challenged the hypothesis that ClO2 decreases the viral load and virus-induced mortality in a vertebrate model. For this, we determined viral load, virus-induced lesions and mortality in 10-day old chick embryos inoculated with 104 mean EID50/mL of attenuated Massachusetts and Connecticut avian coronavirus (IBV) strains.ResultsThe ClO2 treatment had a marked impact on IBV infection. Namely, viral titres were 2.4-fold lower and mortality was reduced by half in infected embryos that were treated with ClO2. Infection led to developmental abnormalities regardless of treatment. Lesions typical of IBV infections were observed in all inoculated embryos, but severity tended to be significantly lower in ClO2-treated embryos. We found no gross or microscopic evidence of toxicity caused by ClO2 at the doses used herein.ConclusionsOur study shows that ClO2 could be a safe and viable way of treating and mitigating the effects of avian coronavirus infections, and raises the possibility that similar effects could be observed in other organisms.Graphical abstract

Summary Background & aims Evening chronotype has been linked with obesity, diabetes and metabolic syndrome (MetS) in middle-aged and older adults. However, few studies have analyzed this association in young adults. The aim of this study was to assess potential associations between individual chronotype and cardiometabolic outcomes in young adults of two independent populations from Europe and America. Methods Total population comprised 2 223 young adults (18–29 years old), 525 from Spain (Europe) and 1 698 from Mexico (America). Anthropometric, body composition and biochemical analyses were performed. Circadian preference was determined using the Morningness-Eveningness Questionnaire (MEQ). Results In these two young adult populations, a higher metabolic risk was found in those individuals with evening chronotypes, whereas those with neither or morning chronotypes showed lower cardiometabolic risk. Evening chronotypes showed lipid alterations with increased levels of triglycerides in both populations, VLDL-c in Spaniards and total cholesterol and LDL-c in Mexicans. Among the Mexican population, evening chronotypes showed higher MetS risk and more obesity traits than the other two chronotypes; no significant differences for the same comparison were found among the equivalent Spanish chronotypes. Evening chronotypes showed lower carbohydrates and higher fat intake in Spaniards, while they had lower fiber intake in Mexicans. The associations between MEQ score and cardiometabolic risk were independent of the dietary characteristics. Lifestyle factors differed among chronotypes with more smokers and habitual drinkers among evening chronotypes than in neither or morning chronotypes (P Conclusions This study performed in two American and European independent populations shows that even in apparently healthy young adults, evening chronotypes have increased cardiometabolic risk and lipid alterations as compared to neither or morning chronotypes.

Although cognitive impairment (CI) is classically associated with aging, it has been proposed that neurological pathologies may increase the risk to suffer CI. Despite the evidence of an elevated prevalence of CI in patients with multiple sclerosis (MS), it is not considered among standard clinical evaluations, due the lack of specialists and time required. The aim of this study was to evaluate if lipid profile is associated with cognitive performance in persons with MS. Twenty patients with MS were evaluated. Montreal Cognitive Assessment (MoCA) was employed to determine cognitive performance. CI was observed in 85% of patients, with memory recall and language as the most affected domains. Despite biomarkers were mostly found within reference values, several correlations were observed. MoCA total score was correlated with cholesterol (r = − 0.468, p = 0.037) and LDL (r = − 0.453, p = 0.045). Visuospatial domain was correlated with LDL (r = − 0.493, p = 0.027). Attention domain correlated with triglycerides (r = − 0.455, p = 0.044) and cholesterol (r = − 0.549, p = 0.012). When the person reaches borderline levels of triglycerides, LDL and cholesterol a decrease in cognitive performance can be observed. The mechanism underlying this association has not been established still, it has been proposed that it could be linked with neuroinflammation, alterations in synapses and in the metabolism of amyloid-β protein. This study settles the potential importance that lipid profile could have on cognitive performance in MS. Further studies are needed to establish optimal levels and implication of lipid profile in the diagnosis and monitoring of cognitive performance in Mexican people with MS.

Lectins are bioactive proteins with the ability to recognize cell membrane carbohydrates in a specific way. Diverse plant lectins have shown diagnostic and therapeutic potential against cancer, and their cytotoxicity against transformed cells is mediated through the induction of apoptosis. Previous works have determined the cytotoxic activity of a Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) and its anti-tumorigenic effect on colon cancer. In this work, lectins from the TBLF were additionally purified by ionic-exchange chromatography. Two peaks with agglutination activity were obtained: one of them was named TBL-IE2 and showed a single protein band in two-dimensional electrophoresis, this one was thus selected for coupling to quantum dot (QD) nanoparticles by microfluidics (TBL-IE2-QD). The microfluidic method led to low sample usage, and resulted in homogeneous complexes, whose visualization was achieved using multiphoton and transmission electron microscopy. The average particle size (380 nm) and the average zeta potential (&minus, 18.51 mV) were determined. The cytotoxicity of the TBL-IE2 and TBL-IE2-QD was assayed on HT-29 colon cancer cells, showing no differences between them (p &le, 0.05), where the LC50 values were 1.0 &times, 10&minus, 3 and 1.7 &times, 3 mg/mL, respectively. The microfluidic technique allowed control of the coupling between the QD and the protein, substantially improving the labelling process, providing a rapid and efficient method that enabled the traceability of lectins. Future studies will focus on the potential use of the QD-labelled lectin to recognize tumor tissues.

Saccharomyces cerevisiae has evolved diverse mechanisms to osmotic changes: the cell wall, ion and water transport systems, and signaling cascades. At the present time, little is known about the mechanisms involved in short-term responses of osmotic stress in yeast or their physiological state during this process. We conducted studies of flow cytometry, wet weight measurements, and electron microscopy to evaluate the modifications in cell volume and the cell wall induced by osmotic stress. In response to osmotic challenges, we show very fast and drastic changes in cell volume (up to 60%), which were completed in less than eight seconds. This dramatic change was completely reversible approximately 16 s after returning to an isosmotic solution. Cell volume changes were also accompanied by adaptations in yeast metabolism observed as a reduction by 50% in the respiratory rate, measured as oxygen consumption. This effect was also fully reversible upon returning to an isosmotic solution. It is noteworthy that we observed a significant recovery in oxygen consumption during the first 10 min of the osmotic shock. The rapid adjustment of the cellular volume may represent an evolutionary advantage, allowing greater flexibility for survival.

The Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) exhibits differential cytotoxicity on colon cancer cells and inhibition of early tumorigenesis in the colon (50 mg/kg, three times per week, for 6 weeks). TBLF showed low toxicity with the ability to activate the immune system; however, some adverse effects are the loss in body weight gain, intestinal atrophy, and pancreatic hyperplasia. After a recovery period of 2 weeks after treatment, reversion of pancreatic hyperplasia but no recovery of intestinal atrophy was observed. As TBLF has shown anticancer effects on the colon, it is important to characterize the adverse effects and how they can be reversed. Sprague Dawley rats were administered with TBLF (50 mg/kg) for 6 weeks, three times per week, and then allowed to recover for 6 weeks post-treatment. After TBLF administration, small intestine atrophy, villus atrophy, and cryptic hyperplasia were confirmed, as well as increased intestinal mucus production, increased permeability and a decrease in the apparent ileal digestibility of crude proteins. The colon showed damage in the simple prismatic tissue and decreased crypt depth, and changes in microbiota and a decrease in the apparent fecal digestibility of crude protein were determined. Our results show that the adverse effects provoked by TBLF were partially reversed after 6 weeks of recovery post-treatment, suggesting that increasing the recovery period it could be possible to reverse all adverse effects observed.

Phenolic compounds are present in goat milk and cheese. The composition of goat milk and its products may vary depending on factors such as season, feeding system and heat treatment. The aim of this work is to quantify total phenolic compounds (TPC) and antioxidant capacity in pasteurized and unpasteurized samples of milk, milk whey, and cheese from goats fed in two different systems (free-range grazing and permanent confinement), during dry and rainy seasons. TPC concentrations were highest in unpasteurized samples from dry season compared to pasteurized and rainy season: 132.4 ± 27.3, 76.5 ± 5.77 mg of gallic acid equivalent (GAE)/L for unpasteurized milk and milk whey, respectively, and 363.21 ± 52.97 mg GAE/Kg for cheese. Antioxidant capacity for dry season produce was significantly higher (P

Graphical abstract, Highlights • TBLF activates immune system through an increase of granulocytes and CD4+ cells. • TBLF provoked activation of Peyer’s patches lymphoid follicles. • TBLF provoked intestinal atrophy with no recovery after two-weeks without treatment. • TBLF provoked exocrine pancreas hypertrophy with a partial recovery. • Spleen did not show significant changes after TBLF treatment., Previous work showed that Tepary bean (Phaseolus acutifolius) lectins exhibit differential cytotoxic effects on cancer cell lines by apoptosis induction. In vivo studies using a Tepary bean lectin fraction (TBLF, 50 mg/kg of body weight) after colon cancer induction in rats showed that TBLF inhibited early precancerous lesions without systemic toxicity however, loss of body weight gain and activation of immune cells were observed. In order to know more about the possible adverse effects, we evaluated the administration of TBLF on digestive and immune organs. Sprague Dawley rats were administered TBLF for six weeks and allowed to recover for two weeks. Immune activation was observed through an increased lymphocyte-granulocyte ratio, an increased number of lymphoid follicles in intestinal Peyer’s patches and a slight expansion of the splenic white pulp. Atrophy was observed in small intestine villi and crypt foci of the colon without normalization after the recovery period. Pancreas histopathology showed hypertrophy after the six-week administration period, particularly vacuolation and trabecular widening; but after the two-week recovery period atrophy was observed, suggesting a partial compensatory type process. Our results show that TBLF activates the immune system and affects digestive organs through direct interaction with intestinal epithelium, and indirectly by producing pancreatic hyperfunction. Further work will focus in longer recuperation periods after TBLF treatment.

Lectins are bioactive proteins with the ability to recognize cell membrane carbohydrates in a specific way. Diverse plant lectins have shown diagnostic and therapeutic potential against cancer, and their cytotoxicity against transformed cells is mediated through the induction of apoptosis. Previous works have determined the cytotoxic activity of a Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) and its anti-tumorigenic effect on colon cancer. In this work, lectins from the TBLF were additionally purified by ionic-exchange chromatography. Two peaks with agglutination activity were obtained: one of them was named TBL-IE2 and showed a single protein band in two-dimensional electrophoresis; this one was thus selected for coupling to quantum dot (QD) nanoparticles by microfluidics (TBL-IE2-QD). The microfluidic method led to low sample usage, and resulted in homogeneous complexes, whose visualization was achieved using multiphoton and transmission electron microscopy. The average particle size (380 nm) and the average zeta potential (−18.51 mV) were determined. The cytotoxicity of the TBL-IE2 and TBL-IE2-QD was assayed on HT-29 colon cancer cells, showing no differences between them (p ≤ 0.05), where the LC50 values were 1.0 × 10−3 and 1.7 × 10−3 mg/mL, respectively. The microfluidic technique allowed control of the coupling between the QD and the protein, substantially improving the labelling process, providing a rapid and efficient method that enabled the traceability of lectins. Future studies will focus on the potential use of the QD-labelled lectin to recognize tumor tissues.

A Tepary bean lectin fraction (TBLF) has been studied because it exhibits differential cytotoxic and anticancer effects on colon cancer. The present work focuses on the evaluation of the apoptotic mechanism of action on colon cancer cells. Initially, lethal concentrations (LC50) were obtained for the three studied cell lines (HT-29, RKO and SW-480). HT-29 showed the highest LC50, 10 and 100 times higher than that of RKO and SW-480 cells, respectively. Apoptosis was evaluated by flow cytometry, where HT-29 cells showed the highest levels of early and total apoptosis, caspases activity was confirmed and necrosis was discarded. The effect on cell cycle arrest was shown in the G0/G1 phase. Specific apoptosis-related gene expression was determined, where an increase in p53 and a decrease in Bcl-2 were observed. Expression of p53 gene showed the maximum level at 8 h with an important decrease at 12 and 24 h, also the phosphorylated p53(ser46) increased at 8 h. Our results show that TBLF induces apoptosis in colon cancer cells by p-p53(ser46) involvement. Further studies will focus on studying the specific signal transduction pathway.

Introduction: aberrant crypt foci (ACF) are colon preneoplastic lesions that can be used as a tool to study preventive processes for colorectal cancer (CRC). This model consists of initiation induced by azoxymethane (AOM) and promoted by sodium dextran sulfate (DSS), simulating human colonic carcinogenesis in a rat model. There is no direct information on the effects of this process on nutritional markers. Objective: to determine the effect on nutritional markers after the induction of ACF by AOM/DSS in a rat model. Methods: ACF were induced in 24 four-week-old Sprague Dawley male rats by administration of 2 AOM injections (10 mg/kg) and 7 days of 2% DSS in their drinking water. Body weight gain, food and fluid intake, weight of sacrificial organs, nutritional biochemical profiles, liver and kidney toxicity were evaluated. Cell counts in blood were also performed and histological sections evaluated in specific organs. The model was confirmed with identification and counts of ACF. Half of the rats were sacrificed at the sub-chronic stage and the rest at the chronic stage. Results: at the sub-chronic stage, changes in the liver and colon weight, and in the lymphocyte count were observed. For both stages, histopathological damage was observed in liver, kidney and colon, along with alterations in serum glucose levels. Conclusions: the model for proposed ACF can be used at the sub-chronic stage without the need for observation at the chronic stage. More research is needed to determine the mechanism of the observed effects.Introducción: los focos de criptas aberrantes (ACF) son lesiones preneoplásicas en colon que pueden ser utilizados como herramienta para estudiar procesos preventivos para el cáncer colorectal (CCR). Este modelo consiste en la iniciación inducida por azoximetano (AOM) y promovida por dextrano sulfato sódico (DSS) simulando una carcinogénesis colónica humana en un modelo de rata. No existe información directa de los efectos sobre marcadores nutricios para este proceso. Objetivo: determinar el efecto sobre marcadores nutricios tras la inducción de ACF por AOM/DSS en un modelo de rata. Métodos: se utilizaron veinticuatro ratas machos Sprague Dawley de 4 semanas para la inducción de ACF por administración de 2 inyecciones de AOM (10 mg/kg) y 7 días de DSS al 2% en el agua para beber. Se evaluó la ganancia de peso corporal, el consumo de alimento y de líquidos, el peso de órganos al sacrificio, perfiles bioquímicos nutricios, de toxicidad hepática y renal. Asimismo, se realizaron conteos celulares en sangre y se evaluaron cortes histológicos en órganos específicos. El modelo se confirmó con la identificación y conteos de ACF. Se sacrificó la mitad de las ratas en etapa subcrónica y las demás en etapa crónica. Resultados: en la etapa subcrónica se observaron cambios entre grupos en el peso del hígado y colon, y en el conteo de linfocitos. En ambas etapas se observaron daños histopatológicos en hígado, riñón y colon, así como alteraciones en los niveles de glucosa sérica. Conclusiones: el modelo para ACF propuesto puede ser utilizado en etapa subcrónica sin necesidad de llevarlo a tiempo crónico. Es necesaria más investigación para determinar el mecanismo de los efectos observados.

Antecedentes y Objetivos: Cnidoscolus aconitifolius es una especie arbustiva que se utiliza como planta ornamental, medicinal y alimentaria en varias partes del mundo, conocida en México como chaya. Existe poca investigación científica publicada sobre su composición fenólica y su capacidad antioxidante. El objetivo de este estudio es determinar la caracterización fenólica y la capacidad antioxidante de extractos hidroalcohólicos (metanol-agua y etanol-agua) en diferentes proporciones de hojas de C. aconitifolius crudas y hervidas.Métodos: Se realizó un cultivo de C. aconitifolius en el campus de la Universidad de Querétaro (México; 2014) utilizando estacas leñosas. Durante el verano de 2016 se realizaron muestreos de hojas de todas las plantas cultivadas. El contenido de compuestos fenólicos totales (CFT), el contenido total de flavonoides (CTF) y la capacidad antioxidante se determinaron en extractos de metanol-agua (50:50 y 80:20) y de etanol-agua (50:50 y 80:20) provenientes de hojas crudas y hervidas. Además, se identificaron y cuantificaron compuestos fenólicos de cada uno de los extractos mediante el método RP-HPLC-DAD.Resultados clave: Se identificaron y cuantificaron 11 compuestos fenólicos en la especie C. aconitifolius: ácido gálico, ácido vanílico, vainillina, ácido clorogénico, ácido cafeico, ácido ferúlico, ácido rosmarínico, ácido p-cumárico, resveratrol, luteolina y apigenina. En general, las hojas hervidas mostraron una mayor capacidad antioxidante que las hojas crudas. Se observaron diferencias en el contenido de compuestos fenólicos debido al tipo de solvente y mezcla utilizada, siendo el extracto etanólico 80:20 proveniente de las hojas hervidas de chaya el que mostró mayor contenido de polifenoles.Conclusiones: Hervir las hojas incrementó el contenido de compuestos fenólicos en los extractos analizados. Los resultados de este estudio indican que las hojas de chaya son una fuente rica de antioxidantes naturales.

Aberrant crypt foci (ACF) is the precursor lesion of colorectal carcinogenesis (CRC), one of the most common malignancies in the world. Many studies have reported that people with higher phytochemical intake are at a reduced risk of developing ACF. One example of the botanical potential of preventive plant products is

Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) has been shown to specifically bind and induce cell death of different types of cancer cells and also has exhibited an effect on early colon tumorigenesis. However, the development of a pharmaceutical formula is not possible yet because the production process is expensive and slow and provides low yields. Therefore, the purpose of the present work was to develop a strategy to produce one bioactive lectin by rhizosecretion through root exudates on genetically modified plants. Amplification of Tepary bean transcripts was performed using degenerate primers, and the products obtained were sequenced. Multiple alignments of sequences led to elucidating one of the lectins present in TBLF. Its coding sequence was flanked by an N-terminal secretion signal peptide and a 6xHis-tail. This construction was introduced into P. acutifolius plants using Agrobacterium tumefaciens to subsequently carry out the in vitro growth of the plants. When roots grew, plants were transferred to hydroponic conditions and root exudates were analyzed. Results showed the presence of a glycosylated cisgenic lectin with biological activity, confirming that the strategy followed provides an alternative for the synthetic production and purification of this lectin.

Overweight/obesity, dyslipidemias, hypertension and hyperglycemia are strongly related to non-communicable diseases (NCD) in which genetic and environmental factors interact with each other. The Mexican population exhibit a genetic disposition to metabolic syndrome, type 2 diabetes, as well as many forms of dyslipidemia. This study aimed to determine the association between biochemical, genetic and environmental factors in the development of metabolic syndrome (MS), obesity and insulin resistance (IR) in Mexican young adults. Young women and men (n = 6750 between 19.3 ± 2.3 years old) participated in a health promotion program from the Autonomous University of Queretaro, Mexico (SU-Salud program). A sub-sample of 665 participants was taken for the determination of single nucleotide polymorphisms (SNP) rs964184 (APOAV), rs9282541 (ABCA1) and rs1260326 (GCKR), using QuantStudio 12 K Flex Real-Time PCR System. For the multivariate analysis, a multiple logistic regression was performed. A prevalence of 22% of overweight and 7% of obesity was determined. The main metabolic risk factors were low levels of HDL-C (30%), IR (19%), and a high level of triglycerides (15%). The main factors associated with IR were body fat percentage and triglycerides; SNP for the ABCA1 gene was related to MS, obesity and low HDL-C; SNP for GCKR gene was related to high fasting glycemia, while APOAV SNP was related with MS, hypertriglyceridemia and low HDL-C. Our findings show that the Mexican genetic predisposition to NCD affects young adults, who can suffer MS, obesity and IR. Public health strategies must focus on prevention actions from an early age.

Tepary bean (Phaseolus acutifolius) lectins exhibit differential in vitro and in vivo biological effects, but their gastrointestinal interactions and digestion have not yet been assessed. This work aimed to evaluate the changes of a recombinant Tepary bean lectin (rTBL-1) through an in vitro and ex vivo gastrointestinal process. A polyclonal antibody was developed to selectively detect rTBL-1 by Western blot (WB) and immunohistochemical analysis. Everted gut sac viability was confirmed until 60 min, where protein bioaccessibility, apparent permeability coefficient, and efflux ratio showed rTBL-1 partial digestion and absorption. Immunoblot assays suggested rTBL-1 internalization, since the lectin was detected in the digestible fraction. The immunohistochemical assay detected rTBL-1 presence at the apical side of the small intestine, potentially due to the interaction with the intestinal cell membrane. The in silico interactions between rTBL-1 and some saccharides or derivatives showed high binding affinity to sialic acid (&minus, 6.70 kcal/mol) and N-acetylglucosamine (&minus, 6.10 kcal/mol). The ultra-high-performance liquid chromatography&ndash, electron spray ionization&ndash, quantitative time-of-flight coupled to mass spectrometry (UHPLC&ndash, ESI&ndash, QTOF/MS) analysis showed rTBL-1 presence in the gastric content and the non-digestible fraction after intestinal simulation conditions. The results indicated that rTBL-1 partially resisted the digestive conditions and interacted with the intestinal membrane, whereas its digestion allowed the absorption or internalization of the protein or the derivative peptides. Further purification of digestion samples should be conducted to identify intact rTBL-1 protein and digested peptides to assess their physiological effects.

Recent findings made by our group indicate that the iron content in Phaseolus vulgaris leaves is at least four times greater than in grains therefore, we evaluated the effect of supplementation with bean leaf (iron content of 275 mg/kg on a dry basis) in iron-deficient rats. Anemia was induced by feeding rats with an iron-deficient diet (IDD) for 11 days and iron-recovery diets were subsequently tested for 14 days using a normal diet, a 10 % bean leaf-supplemented IDD (BLSD) or a ferrous sulfate-supplemented IDD. Decreased levels of leukocytes (64 %), erythrocytes (30 %), lymphocytes (62 %), granulocytes (72 %), hematocrit (34 %), hemoglobin (35 %), and ferritin (34 %) were observed in the iron-deficient rats compared to the control rats. BLSD supplementation showed the highest recovery values relative to those recorded for control rats: leukocytes (40 %), erythrocytes (24 %), lymphocytes (33 %), granulocytes (88 %), hematocrit (17 %), and hemoglobin (18 %), suggesting that common bean leaves could be a good source of bioavailable iron with possible immunomodulatory effects.