Risk of chronic diseases in indigenous Totonacs from Mexico short running title: Risk of chronic diseases in indigenous Totonacs.

AIM: The purpose of this study was to determine the risk of noncommunicable diseases (NCDs) development in four indigenous Totonac communities. BACKGROUND: Poverty and low education levels increase the risk of unhealthy lifestyles, leading to a higher incidence of NCDs in indigenous communities. In addition, limited access to health services significantly reduces the opportune screening of risk factors. METHODS: This was a cross-sectional observational study, in which we evaluated the incidence and/or the risk (%) to develop NCDs in adults from indigenous Totonac communities of Puebla State, Mexico. The sample consisted of 255 adults over 20 years old. Analysis of variance (ANOVA), Student's t-test, and Pearson's correlation were used for statistical analysis. RESULTS: We found significant differences between communities regarding the risk of diabetes (p < 0.05) and hypertension (p < 0.01). However, there were no significant differences in terms of CVD risk (p > 0.05). Men were at higher risk for hypertension and CVD than women (37.8% vs 27.3% and 20.9% vs 12.2%, respectively), whereas women were at a higher risk of diabetes than men based on the FINDRISC score (11.7 vs 9.7, respectively). CONCLUSION: The Zapotitlan community showed a higher risk of developing both diabetes and hypertension compared with the other communities. Men showed higher risks of hypertension and CVD compared with women. Women are at higher risk of diabetes than men.

Autophagy Activation by Resveratrol Reduces Severity of Experimental Rheumatoid Arthritis.

SCOPE: Previous work reported that dietary supplementation with resveratrol lowers synovial hyperplasia, inflammatory and oxidative damage in an antigen-induced arthritis (AIA) model. Here, it is investigated whether resveratrol can regulate the abnormal synovial proliferation by inducing autophagy and controlling the associated inflammatory response. METHODS AND RESULTS: Animals treated with resveratrol 8 weeks before AIA induction show the highest significant signal for microtubule-associated protein 1 light chain 3 by confocal microscopy. Besides, resveratrol significantly reduces p62 expression, but it does not increase the signal of beclin-1. Also, active caspase-3 expression, as well as poly(ADP-ribose) polymerase, is upregulated in the AIA group, and is significantly reduced in resveratrol-treated AIA group. Resveratrol also mitigates angiopoietin-1 and vascular endothelial growth factor signals. Finally, resveratrol significantly reduces the serum levels of IL-1ß, C reactive protein, and prostaglandin E2, as well as nuclear factor κB synovial tissue expression, which shows a significant correlation with p62 expression. CONCLUSION: Dietary supplementation with resveratrol induces the noncanonical autophagy pathway and limits the cross-talk with inflammation, which in consequence modulates the synovial hyperplasia. Preventive strategies that incorporate dietary intervention with resveratrol may offer a potential therapeutic alternative to drugs to influence the risk of rheumatoid arthritis and influence its course.

Influencia de la lengua dispar (tutunakú y español) en los mensajes e instrucciones que emite el personal de salud y la de las receptoras de la información / Influence of disparate language (Tutunakú and Spanish) in messages and instructions issued by health personnel and those receiving the information

Resumen OBJETIVO: Identificar la influencia de la lengua dispar (tutunakú y español) entre los mensajes e instrucciones que emite el personal de salud y la de las receptoras de esa información, en particular la dirigida a la práctica de la autoexploración con fines de prevención del cáncer de mama. MATERIALES Y MÉTODOS: Estudio descriptivo, transversal, efectuado en mujeres indígenas, mayores de 20 años, a quienes se aplicó un instrumento semiestructurado para la identificación de factores de riesgo y protección de cáncer de mama. Para determinar la asociación de las variables de hablantes de totonakú con el nivel de conocimiento, prevención y riesgo de cáncer de mama se utilizó la prueba de χ2. RESULTADOS: Se estudiaron 187 mujeres; de las que 109 (58.5%) solo hablaban tutunakú y 78 (41.5%) eran bilingües (tutunakú y español). La edad promedio de las participantes fue de 43.4 años (DE ± 14.8), con límites de 20 y 80 años. En relación con la escolaridad: 93 de 109 (85.5%) de las monolingües eran analfabetas y solo 44 de las 78 (57.1%) mujeres bilingües habían concluido la educación básica. Por lo que se refiere al conocimiento del cáncer de mama, las mujeres monolingües tuvieron menor nivel de conocimiento y menor práctica de la autoexploración; solo 34 de las 78 bilingües practicaban la autoexploración. CONCLUSIÓN: Si bien el riesgo de cáncer de mama no muestra asociación con el hecho de hablar una lengua sí lo tiene, directamente, con la escolaridad, con la capacidad necesaria para comprender instrucciones que permitan la práctica de la autoexploración.

Abstract OBJECTIVE: To identify the influence of the disparate language (tutunaku and Spanish) between the messages and instructions issued by health personnel and those of the recipients of this information, in particular that directed at the practice of self-examination for the purpose of breast cancer prevention. MATERIALS AND METHODS: Descriptive, cross-sectional study, carried out on indigenous women over 20 years of age, to whom a semi-structured instrument was applied for the identification of risk factors and protection from breast cancer. To determine the association of Totonaku speakers' variables with the level of knowledge, prevention and risk of breast cancer, the test of 2 was used. RESULTS: A total of 187 women were studied; 109 (58.5%) spoke only tutunaku and 78 (41.5%) were bilingual (tutunaku and Spanish). The average age of participants was 43.4 years (SD 14.8), with limits of 20 and 80 years. With regard to schooling: 93 of 109 (85.5%) of the monolingual women were illiterate and only 44 of 78 (57.1%) of the bilingual women had completed basic education. With regard to knowledge of breast cancer, monolingual women had a lower level of knowledge and less practice of self-examination; only 34 of the 78 bilingual women practiced self-examination. CONCLUSION: Although the risk of breast cancer is not associated with speaking a language, it is directly associated with schooling, with the ability to understand instructions that allow the practice of self-examination.

Genetic abrogation of the fibronectin-α5ß1 integrin interaction in articular cartilage aggravates osteoarthritis in mice.

The balance between synthesis and degradation of the cartilage extracellular matrix is severely altered in osteoarthritis, where degradation predominates. One reason for this imbalance is believed to be due to the ligation of the α5ß1 integrin, the classic fibronectin (FN) receptor, with soluble FN fragments instead of insoluble FN fibrils, which induces matrix metalloproteinase (MMP) expression. Our objective was to determine whether the lack of α5ß1-FN binding influences cartilage morphogenesis in vivo and whether non-ligated α5ß1 protects or aggravates the course of osteoarthritis in mice. We engineered mice (Col2a-Cre;Fn1RGE/fl), whose chondrocytes express an α5ß1 binding-deficient FN, by substituting the aspartic acid of the RGD cell-binding motif with a glutamic acid (FN-RGE). At an age of 5 months the knee joints were stressed either by forced exercise (moderate mechanical load) or by partially resecting the meniscus followed by forced exercise (high mechanical load). Sections of femoral articular knees were analysed by Safranin-O staining and by immunofluorescence to determine tissue morphology, extracellular matrix proteins and matrix metalloproteinase expression. The articular cartilage from untrained control and Col2a-Cre;Fn1RGE/fl mice was normal, while the exposure to high mechanical load induced osteoarthritis characterized by proteoglycan and collagen type II loss. In the Col2a-Cre;Fn1RGE/fl articular cartilage osteoarthritis progressed significantly faster than in wild type mice. Mechanistically, we observed increased expression of MMP-13 and MMP-3 metalloproteinases in FN-RGE expressing articular cartilage, which severely affected matrix remodelling. Our results underscore the critical role of FN-α5ß1 adhesion as ECM sensor in circumstances of articular cartilage regeneration.

Resveratrol lowers synovial hyperplasia, inflammatory markers and oxidative damage in an acute antigen-induced arthritis model.

OBJECTIVE: The present study aimed to determine the protective effects of dietary supplementation with resveratrol (RSV) in an acute antigen-induced arthritis (AIA) model. METHODS: Rats were randomly divided into three groups: control, AIA and RSV-treated AIA group. RSV (12.5 mg/kg/day) was given orally for 8 weeks before induction of AIA and until the end of the experiment (48 h after intra-articular injection). The control and AIA animals were administered 100 µl of water. Results were evaluated by macroscopic observation, histopathology and immunohistochemistry for anti-PCNA, macrophages (CD68), T lymphocytes (CD3), monocyte chemoattractant protein-1 and 8-oxo-7,8-dihydro-2'-deoxyguanine (a marker of DNA damage). Cytokine-induced neutrophil chemoattractant-1 in serum and peroxidase activity in synovial tissue were measured using commercial kits. RESULTS: At the end of the study, RSV significantly reduced knee swelling. Likewise, the histological score of synovial tissue also reduced significantly. The arthritis-protective effects were associated with a significant decrease in PCNA, CD68, CD3 and monocyte chemoattractant protein-1 staining, as well as a reduction in serum concentrations of cytokine-induced neutrophil chemoattractant-1. RSV treatment also decreased the level of the marker of DNA damage, 8-oxo-7,8-dihydro-2'-deoxyguanine. Accordingly, peroxidase activity in the synovial tissue was up-regulated. CONCLUSION: Dietary supplementation with RSV lowers the main pathological hallmarks of RA disease in an acute model of AIA. RSV may represent a promising strategy in controlling the severity of RA.

Changes in the integrins expression are related with the osteoarthritis severity in an experimental animal model in rats.

We identify changes in the expression and localization of α5, α4, and α2 integrins during osteoarthritis (OA) pathogenesis in a rat experimental model. The changes were concomitant with variations in the extracellular matrix (ECM) content and the increase of metalloproteinases (MMPs) activity during OA pathogenesis, which were analyzed by immunofluorescence and Western blot assays. Our results showed an increased expression of α5 and α2 integrins at OA late stages, which was co-related with changes in the ECM content, as a consequence of the MMPs activity. In addition, this is the first report that has shown the presence of α4 integrin since OA early stages, which was co-related with the loss of proteoglycans and clusters formation. However, at late OA stages, the increased expression of α4 integrin in the middle and deep zones of the cartilage was also co-related with the abnormal endochondral ossification of the cartilage through its interaction with osteopontin. Finally, we conclude that ECM-chondrocytes interaction through specific cell receptors is essential to maintain the cartilage homeostasis. However, due to integrins cell signaling is ligand-dependent; changes in the ECM contents could induce activation of either anabolic or catabolic processes, which limits the reparative capacity of chondrocytes, favoring OA severity.

Hepatocyte growth factor protects against isoniazid/rifampicin-induced oxidative liver damage.

The worldwide increment of multidrug- and extensively drug-resistant tuberculosis has emphasized the importance of looking for new options in therapeutics. Long-time usage or higher doses of isoniazid and rifampicin have been considered for the treatment of multidrug-resistant tuberculosis; however, the risk of liver failure is proportionally increased. Hepatocyte growth factor (HGF) is a multitask growth factor that stimulates both antiapoptotic and antioxidant responses that counteract the toxic effects of drug metabolism in the liver. The present work was focused to address the antioxidant and antiapoptotic effects of HGF on isoniazid- and rifampicin-induced hepatotoxicity. BALB/c mice were subjected to rifampicin (150mg/kg, intragavage [ig]) plus isoniazid (75mg/kg, ig) for 7 days. Increments in alanine aminotransferase activity, steatosis, apoptosis, and oxidative stress markers were found in animals. Recombinant HGF (iv) prevented all the harmful effects by increasing the activation of Erk1/2 and PKCδ signaling pathways and glutathione (GSH) synthesis. Furthermore, inhibition of endogenous HGF with anti-HGF antibody (iv) enhanced the isoniazid- and rifampicin-induced oxidative stress damage and decreased the GSH content, aggravating liver damage. In conclusion, HGF demonstrated to be a good protective factor against antituberculosis drug-induced hepatotoxicity and could be considered a good adjuvant factor for the use of high doses of or the reintroduction of these antituberculosis drugs.

Perlecan maintains microvessel integrity in vivo and modulates their formation in vitro.

Perlecan is a heparan sulfate proteoglycan assembled into the vascular basement membranes (BMs) during vasculogenesis. In the present study we have investigated vessel formation in mice, teratomas and embryoid bodies (EBs) in the absence of perlecan. We found that perlecan was dispensable for blood vessel formation and maturation until embryonic day (E) 12.5. At later stages of development 40% of mutant embryos showed dilated microvessels in brain and skin, which ruptured and led to severe bleedings. Surprisingly, teratomas derived from perlecan-null ES cells showed efficient contribution of perlecan-deficient endothelial cells to an apparently normal tumor vasculature. However, in perlecan-deficient EBs the area occupied by an endothelial network and the number of vessel branches were significantly diminished. Addition of FGF-2 but not VEGF(165) rescued the in vitro deficiency of the mutant ES cells. Furthermore, in the absence of perlecan in the EB matrix lower levels of FGFs are bound, stored and available for cell surface presentation. Altogether these findings suggest that perlecan supports the maintenance of brain and skin subendothelial BMs and promotes vasculo- and angiogenesis by modulating FGF-2 function.

Guidelines for the use and interpretation of assays for monitoring autophagy.

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.