Preliminary Analysis of Unsaturated Fatty Acid Profiles of Coffea arabica L., in Samples with a Denomination of Origin and Speciality of Oaxaca, Mexico.

In February 2020, Coffea arabica L. grown on the coast and in the Southern Sierra Madre of the state of Oaxaca, Mexico obtained the denomination of origin. Which does not have data on color and chemical composition, the first associated with the degree of roasting and the second with lipids (17-18%), as the group of compounds responsible, in part, for flavor, consistency, and may contribute to health benefits. In the present work, color was determined on the CIE L*a*b* scale and the unsaturated fatty acids by Nuclear Magnetic Resonance (NMR) of 1H and 13C in samples of medium roasted specialty coffee from the "Pluma" coffee-growing region, Oaxaca, Mexico. The average value of L* luminosity in ground coffee was 42.1 ± 0.1 reported for a light roast. Unsaturated fatty acids were quantified from the lipid fraction of the gr1 ound grain by NMR 1H and 13C, obtaining on average the highest abundance of linoleic (41.7 ± 0.5 by 1 H and 41.24 ± 0.5 by 13C), followed by oleic (9.2 ± 0.2 by 1H and 7.4 ± 0.2 by 13C) and linolenic (1.5 ± 0.1 by H and 1.1 ± 0.2 by 13C). This study indicates that 1H and 13C NMR spectroscopy is a useful tool for the quantification of linolenic, linoleic, and oleic fatty acids by the method of key signal shifts of these acids found in lipid samples in roasted coffee grains.

The Role of B Cell and T Cell Glycosylation in Systemic Lupus Erythematosus.

Glycosylation is a post-translational modification that affects the stability, structure, antigenicity and charge of proteins. In the immune system, glycosylation is involved in the regulation of ligand-receptor interactions, such as in B-cell and T-cell activating receptors. Alterations in glycosylation have been described in several autoimmune diseases, such as systemic lupus erythematosus (SLE), in which alterations have been found mainly in the glycosylation of B lymphocytes, T lymphocytes and immunoglobulins. In immunoglobulin G of lupus patients, a decrease in galactosylation, sialylation, and nucleotide fucose, as well as an increase in the N-acetylglucosamine bisector, are observed. These changes in glycoisolation affect the interactions of immunoglobulins with Fc receptors and are associated with pericarditis, proteinuria, nephritis, and the presence of antinuclear antibodies. In T cells, alterations have been described in the glycosylation of receptors involved in activation, such as the T cell receptor; these changes affect the affinity with their ligands and modulate the binding to endogenous lectins such as galectins. In T cells from lupus patients, a decrease in galectin 1 binding is observed, which could favor activation and reduce apoptosis. Furthermore, these alterations in glycosylation correlate with disease activity and clinical manifestations, and thus have potential use as biomarkers. In this review, we summarize findings on glycosylation alterations in SLE and how they relate to immune system defects and their clinical manifestations.

Mechanisms of Immunothrombosis by SARS-CoV-2.

SARS-CoV-2 contains certain molecules that are related to the presence of immunothrombosis. Here, we review the pathogen and damage-associated molecular patterns. We also study the imbalance of different molecules participating in immunothrombosis, such as tissue factor, factors of the contact system, histones, and the role of cells, such as endothelial cells, platelets, and neutrophil extracellular traps. Regarding the pathogenetic mechanism, we discuss clinical trials, case-control studies, comparative and translational studies, and observational studies of regulatory or inhibitory molecules, more specifically, extracellular DNA and RNA, histones, sensors for RNA and DNA, as well as heparin and heparinoids. Overall, it appears that a network of cells and molecules identified in this axis is simultaneously but differentially affecting patients at different stages of COVID-19, and this is characterized by endothelial damage, microthrombosis, and inflammation.

Evaluación del crecimiento radial de Lactarius volemus en medio semisólido / Evaluation of radial growth of Lactarius volemus in semisolid medium

Objetivo: Evaluar el crecimiento radial de Lactarius volemus en cinco medios de cultivo semisólidos in vitro.Materiales y métodos: Cuerpos fructíferos de L. volemus provenientes de la Sierra Norte del estado de Oaxaca, México, se cultivaron en laboratorio en medios Agar Papa Dextrosa, Agar Czapek-Dox, Agar Extracto de Malta, Agar Papa Sacarosa y Agar Dextrosa Saboraud; mediante dos técnicas de sembrado. Se evaluaron las características morfológicas de colonias obtenidas de distintas muestras del cuerpo fructífero, así como el crecimiento radial de cada una.Resultados: El crecimiento colonial evaluado permitió seleccionar un medio que reúne las condiciones óptimas para el cultivo de Lactarius volemus in vitro. No todas las muestras utilizadas desarrollan un crecimiento abundante: la muestra proveniente del látex presenta un crecimiento escaso.Conclusiones: Con la evaluación del crecimiento radial de Lactarius volemus se obtiene una referencia directa del ciclo de crecimiento de esta especie; es posible identificar las fases exponencial y estacionaria pero las condiciones del medio no permiten evaluar la fase de muerte debido a la deshidratación y reducción del agar.(AU)

Objective: To evaluate the radial growth of Lactarius volemus in five semi-solid culture media in vitro.Materials and methods: Fruitful bodies of L. volemus from the Sierra Norte of the state of Oaxaca, Mexico, were cultured in the laboratory in Potato Dextrose Agar Papa, Czapek-Dox Agar, Malt Extract Agar, Potato Sucrose Agar and Dextrose Saboraud Agar; using two seeding techniques. The morphological characteristics of colonies obtained from different samples of the fruiting body were evaluated, as well as the radial growth of each one.Results: The evaluated colonial growth allowed to select a culture medium that meets the optimal conditions for the cultivation of Lactarius volemus in vitro. Not all samples used develop abundant growth: the sample from latex shows little growth.Conclusions: With the evaluation of the radial growth of Lactarius volemus a direct reference to the growth cycle of this species is obtained; it is possible to identify the exponential and stationary phases but the conditions of the medium do not allow evaluating the phase of death due to dehydration and reduction of the agar.(AU)

Cuantificación de la actividad específica de lectina en teosinte Zea diploperennis / Quantification of specific lectin activity in teosinte Zea diploperennis

Objetivo: Evaluar la actividad específica de lectina en teosinte Zea diploperennis sano e infectado con Ustilago maydis.Materiales y métodos: Plántulas de Zea diploperennis de 6 días de crecimiento fueron inoculadas con Ustilago maydis. Se evaluó la concentración de proteínas totales y la actividad hemaglutinante de extractos crudos de teosinte sano e infectado en placas de microtitulación con eritrocitos tipo O al 3% siguiendo la técnica de diluciones dobles seriadas durante 6 días. Resultados: La concentración de proteínas totales se incrementa en coleoptilo sano durante cada día de su crecimiento. No así en teosinte infectado donde la curva presenta una tendencia a la baja desde el momento de la inoculación. La actividad específica de lectina disminuye en ambos casos desde el primer día de cuantificación. Conclusiones: La evidente reducción en la actividad de lectina en teosinte infectado en comparación con teosinte sano podría explicar la susceptibilidad de este teosinte a dicho fitopatógeno. Si bien, la participación de las lectinas de teosinte y maíz en el mecanismo de defensa a Ustilago maydis todavía no ha sido esclarecida, los resultados obtenidos contribuyen a la comprensión del efecto que puede tener la concentración de lectina y proteína sobre la resistencia en teosinte.(AU)

Objective: To evaluate the specific activity of lectin in healthy and infected teosinte Zea diploperennis with Ustilago maydis. Materials and methods: Zea diploperennis seedlings of 6 days of growth were inoculated with Ustilago maydis. Total protein concentration and hemagglutinating activity of crude extracts of healthy and infected teosinte were evaluated in microtiter plates with 3% type O red cells following the technique of serial double dilutions for 6 days. Results: The concentration of total proteins increases in healthy coleoptile during each day of its growth. Not so in infected teosinte where the curve shows a downward trend from the moment of inoculation. The specific lectin activity decreases in both cases from the first day of quantification. Conclusions: The evident reduction in lectin activity in infected teosinte compared to healthy teosinte could explain the susceptibility of this teosinte to said phytopathogen. Although the participation of the teosinte and corn lectins in the defense mechanism against Ustilago maydis has not yet been clarified, the results obtained contribute to the understanding of the effect that the concentration of lectin and protein can have on resistance in teosinte.(AU)

Analysis of SARS-CoV-2 mutations in Mexico, Belize, and isolated regions of Guatemala and its implication in the diagnosis.

The genomic sequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide are publicly available and are derived from studies due to the increase in the number of cases. The importance of study of mutations is related to the possible virulence and diagnosis of SARS-CoV-2. To identify circulating mutations present in SARS-CoV-2 genomic sequences in Mexico, Belize, and Guatemala to find out if the same strain spread to the south, and analyze the specificity of the primers used for diagnosis in these samples. Twenty three complete SARS-CoV-2 genomic sequences, available in the GISAID database from May 8 to September 11, 2020 were analyzed and aligned versus the genomic sequence reported in Wuhan, China (NC_045512.2), using Clustal Omega. Open reading frames were translated using the ExPASy Translate Tool and UCSF Chimera (v.1.12) for amino acid substitutions analysis. Finally, the sequences were aligned versus primers used in the diagnosis of COVID-19. One hundred and eighty seven distinct variants were identified, of which 102 are missense, 66 synonymous and 19 noncoding. P4715L and P5828L substitutions in replicase polyprotein were found, as well as D614G in spike protein and L84S in ORF8 in Mexico, Belize, and Guatemala. The primers design by CDC of United States showed a positive E value. The genomic sequences of SARS-CoV-2 in Mexico, Belize, and Guatemala present similar mutations related to a virulent strain of greater infectivity, which could mean a greater capacity for inclusion in the host genome and be related to an increased spread of the virus in these countries, furthermore, its diagnosis would be affected.

Vitis vinifera: An Alternative for the Prevention of Neurodegenerative Diseases.

To present a systematic review of published studies in databases such as PUBMED, REDALYC, SCIELO, DIALNET, SCOPUS, EBSCO and CONRICYT related to the role-played by the components present in the vegetable oil of grape seed (Vitis vinífera) and the prevention or delay in the onset or progression of neurodegenerative diseases. The analysis of the research revealed that neurodegenerative diseases causes alterations in consciousness or in the nervous system leading to severe damage in neuronal cells, these pathologies are considered gradual and progressive. Various syndromes manifest the degenerative diseases of the nervous system; in some of them the predominant symptom is the progressive dementia. Among the components of the diet that in numerous epidemiological studies have shown an inverse association are vitamins, minerals, carotenoids, polyunsaturated fatty acids and polyphenols, the latter being the ones addressed in this document. There is an important evidence that a nutritional support based on polyunsaturated fatty acids and antioxidants can be applied to subjects with a history of neurodegenerative conditions in order to act as neuroprotectors. This requires the determination of the nutritional benefits of these nutrients or of nutraceuticals for the health of this group of patients.

TSH Levels in Subclinical Hypothyroidism in the 97.5th Percentile of the Population.

The debate regarding the cutoff point in the treatment of patients with subclinical hypothyroidism (Shypo) is ongoing. Generally, two different groups are identified for treatment by levels of 10 and 20 mIU/L. Nevertheless, the question remains, "what cutoff point should be chosen?" We have written a selective nonsystematic review focused on the 97.5 percentile reference value reported in healthy subjects in a number of countries and observed important disparities, which partly show the challenge of identifying a single cutoff point for those patients needing medication. We identified studies of TSH on the natural history of subclinical hypothyroidism from population-based prospective cohort studies, which follow up patients for several years. The evolution of TSH levels in these patients is variable. Some cases of TSH may return to lower levels at different stages over the years, but others may not, possibly even developing into overt thyroid failure, also variable. We analyzed factors that may explain the normalization of serum TSH levels. In addition, we found that thorough population-based prospective cohort studies following up on TSH levels, thyroid antibodies, and ultrasonography are important in decisions made in the treatment of patients. However, the 97.5 percentile reference value varies in different countries; therefore, an international cutoff point for subclinical hypothyroidism cannot be recommended.

Aggregation and Molecular Properties of ß-Glucosidase Isoform II in Chayote (Sechium edule).

The presence of isoforms of ß-glucosidase has been reported in some grasses such as sorghum, rice and maize. This work aims to extract and characterize isoform II in ß-glucosidase from S. edule. A crude extract was prepared without buffer solution and adjusted to pH 4.6. Contaminating proteins were precipitated at 4 °C for 24 h. The supernatant was purified by chromatography on carboxymethyl cellulose (CMC) column, molecular exclusion on Sephacryl S-200HR, and exchange anionic on QFF column. Electrophoretic analyzes revealed a purified enzyme with aggregating molecular complex on SDS-PAGE, Native-PAGE, and AU-PAGE. Twelve peptides fragments were identified by nano liquid chromatography-tandem mass spectrometry (nano LC-ESI-MS/MS), which presented as 61% identical to Cucurbita moschata ß-glucosidase and 55.74% identical to ß-glucosidase from Cucumis sativus, another Cucurbitaceous member. The relative masses which contained 39% hydrophobic amino acids ranged from 982.49 to 2,781.26. The enzyme showed a specificity to ß-d-glucose with a Km of 4.59 mM, a Vmax value of 104.3 µM∙min-1 and a kcat of 10,087 µM∙min-1 using p-nitrophenyl-ß-D-glucopyranoside. The presence of molecular aggregates can be attributed to non-polar amino acids. This property is not mediated by a ß-glucosidase aggregating factor (BGAF) as in grasses (maize and sorghum). The role of these aggregates is discussed.

Obesity subtypes, related biomarkers & heterogeneity.

Obesity is a serious medical condition worldwide, which needs new approaches and recognized international consensus in treating diseases leading to morbidity. The aim of this review was to examine heterogeneous links among the various phenotypes of obesity in adults. Proteins and associated genes in each group were analysed to differentiate between biomarkers. A variety of terms for classification and characterization within this pathology are currently in use; however, there is no clear consensus in terminology. The most significant groups reviewed include metabolically healthy obese, metabolically abnormal obese, metabolically abnormal, normal weight and sarcopenic obese. These phenotypes do not define particular genotypes or epigenetic gene regulation, or proteins related to inflammation. There are many other genes linked to obesity, though the value of screening all of those for diagnosis has low predictive results, as there are no significant biomarkers. It is important to establish a consensus in the terminology used and the characteristics attributed to obesity subtypes. The identification of specific molecular biomarkers is also required for better diagnosis in subtypes of obesity.

A preliminary study of platelet hyperactivity in the chronic indeterminate phase of Chagas' disease.

The chronic indeterminate phase of Chagas' disease is asymptomatic despite positive test results for antibodies specific to Trypanosoma cruzi. CD62P-APC (P-selectin) and PAC-1 FITC (GpIIb/IIIa) may improve diagnosis as biomarkers of platelet activity. Nine asymptomatic seropositive subjects, previously untreated, were selected from a blood bank within a year of Chagas' disease detection, in addition to a control group of four. All subjects were evaluated by flow cytometry for CD62P, PAC-1 and CD41, and in a complementary study, by Tissue Doppler Echocardiography for isovolumic relaxation times (IVRT) and E/A ratios. The subjects were classified as positive or negative for CD62P and PAC-1 by a cut off obtained from their mean±2SD. For IVRT and E/A ratios, cut offs were obtained from the American Society of Echocardiography and the European Association of Cardiovascular Imaging recommendations. Fisher's exact test was used for associated findings. Pre-test and post-test probability, sensitivity, specificity, positive and negative predictive values and likelihood ratios were calculated. Abnormalities were expressed as platelet hyperactivity and ventricular dysfunction in CD62P, PAC-1, IVRT and E/A ratios. CD62P appears to have greater sensitivity (0.75) and PAC-1, more accurate specificity (0.75), which may explain thrombotic events in Chagas' disease. We recommend the use of CD62P and PAC-1 as biomarkers of platelet hyperactivity in patients in the chronic indeterminate phase of Chagas' disease.

Reduced platelet aggregation in women after intercourse: a possible role for the cyclooxygenase pathway.

We hypothesise that molecules in the cyclooxygenase pathway affect platelet activity when seminal fluid (SF) is present. We considered the influence of SF on platelet aggregation in women, and believe that the prostanoids in SF signalling are significant. Thirty-one female subjects were studied, 20 of whom were sexually active. Male partners were given either aspirin or indomethacin to inhibit cyclooxygenase. The 6-keto prostaglandin F1α (6-keto PGF1α) and prostaglandin E metabolite (PGE-M) in SF were measured by competitive assay. Platelets and prostanoids were evaluated in women, periodically, before and after intercourse. The platelets were tested with adenosine diphosphate (ADP) and arachidonic acid (AA). To block the interaction between the uterus and SF, some couples used condoms. We found that the 6-keto prostaglandin F1α in urine at 2 hours post-intercourse (1418.75 pg/mL, Std 688.39) was greater than pre-intercourse (772.68 pg/mL, Std 116.54). Post-intercourse, a transient decrease in platelet aggregation was observed in women whose partners did not use condoms. Averages for platelet aggregation were 20.16% with ADP, and more significantly, 37.79% with AA after 2 hours. In contrast, couples using condoms showed no changes, averaging 64.02% with ADP and 72.06% with AA. Women whose partners were taking aspirin or indomethacin also showed no changes. SF from men taking aspirin or indomethacin led to no reduction in platelet aggregometry in their partners. These results indicate that in cases of exposure to SF, the transient change in women's platelet activity could be related to the cyclooxygenase pathway.

Protein C activation peptide inhibits the expression of ICAM-1, VCAM-1, and interleukin-8 induced by TNF-α in human dermal microvascular endothelial cells.

Activated protein C (APC) is generated from the cleavage of protein C by thrombin coupled to thrombomodulin and, subsequently, is released as protein C activation peptide (papC). The aim of this study was to evaluate the effect of papC on human dermal microvascular endothelial cells (HMEC-1), activated with 5 ng//mL TNF-α. Flow cytometry showed that papC inhibited the expression of VCAM-1 and ICAM-1, after activation with TNF-a. Similarly, RT-PCR analysis revealed that 2 and 4 pM papC inhibited the expression of VCAM-1 and IL-8 mRNA in TNF-α-treated HMEC-1. In addition, the expression of endothelial nitric oxide synthase(eNOS) increased in HMEC-1 treated with papC, compared to those without treatment. Furthermore, Jurkat cell adhesion to HMEC-1 induced by TNF-a was significantly inhibited after the addition of papC, compared to HMEC-1 without papC (p = 0.03). Finally, a control peptide analog to papC showed no effect on the expression of ICAM and VCAM on the surface of HMEC-1. In conclusion, our results suggest that papC exerts anti-inflammatory effects on endothelial cells.