Cordycepin Triphosphate as a Potential Modulator of Cellular Plasticity in Cancer via cAMP-Dependent Pathways: An In Silico Approach.

Cordycepin, or 3'-deoxyadenosine, is an adenosine analog with a broad spectrum of biological activity. The key structural difference between cordycepin and adenosine lies in the absence of a hydroxyl group at the 3' position of the ribose ring. Upon administration, cordycepin can undergo an enzymatic transformation in specific tissues, forming cordycepin triphosphate. In this study, we conducted a comprehensive analysis of the structural features of cordycepin and its derivatives, contrasting them with endogenous purine-based metabolites using chemoinformatics and bioinformatics tools in addition to molecular dynamics simulations. We tested the hypothesis that cordycepin triphosphate could bind to the active site of the adenylate cyclase enzyme. The outcomes of our molecular dynamics simulations revealed scores that are comparable to, and superior to, those of adenosine triphosphate (ATP), the endogenous ligand. This interaction could reduce the production of cyclic adenosine monophosphate (cAMP) by acting as a pseudo-ATP that lacks a hydroxyl group at the 3' position, essential to carry out nucleotide cyclization. We discuss the implications in the context of the plasticity of cancer and other cells within the tumor microenvironment, such as cancer-associated fibroblast, endothelial, and immune cells. This interaction could awaken antitumor immunity by preventing phenotypic changes in the immune cells driven by sustained cAMP signaling. The last could be an unreported molecular mechanism that helps to explain more details about cordycepin's mechanism of action.

Breast Cancer Tissue Explants: An Approach to Develop Personalized Therapy in Public Health Services.

Breast cancer is one of the main causes of death worldwide. Lately, there is great interest in developing methods that assess individual sensitivity and/or resistance of tumors to antineoplastics to provide personalized therapy for patients. In this study we used organotypic culture of human breast tumor slices to predict the experimental effect of antineoplastics on the viability of tumoral tissue. Samples of breast tumor were taken from 27 patients with clinically advanced breast cancer; slices were obtained and incubated separately for 48 h with paclitaxel, docetaxel, epirubicin, 5-fluorouracil, cyclophosphamide, and cell culture media (control). We determined an experimental tumor sensitivity/resistance (S/R) profile by evaluating tissue viability using the Alamar Blue® metabolic test, and by structural viability (histopathological analyses, necrosis, and inflammation). These parameters were related to immunohistochemical expression of the estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. The predominant histological type found was infiltrating ductal carcinoma (85.2%), followed by lobular carcinoma (7.4%) and mixed carcinoma (7.4%). Experimental drug resistance was related to positive hormone receptor status in 83% of samples treated with cyclophosphamide (p = 0.027). Results suggest that the tumor S/R profile can help to predict personalized therapy or optimize chemotherapeutic treatments in breast cancer.

[Educational intervention on organ and tissue donation at UMAE No. 34]. / Intervención educativa sobre donación de órganos y tejidos en la UMAE No. 34.

Background: The lack of information associated with donation is devastating for those patients in need of a transplant and requires a solution based on changing social perception through educational interventions. Objective: Determine the level of knowledge of the general population after an educational intervention about organ and tissue donation at the Hospital de Cardiología UMAE No. 34. Methods: Educational intervention study with measurement before and after, prospective. Instrument validated using the Kuder-Richardson formula with a reliability coefficient of 0.74. The study population was made up of the general population in the waiting rooms at UMAE No. 34, only the associations with values of p < 0.05 were considered statistically significant. Results: 266 evaluation instruments were applied to 133 participants. The educational intervention contributed to an increase in the level of knowledge (p = 0.0001). The level of knowledge after the intervention was higher in the younger participants (p = 0.013) and in those with a university studies (p = 0.0001). The relation between age and the level of subsequent knowledge showed favorable significance in the intention to donate in younger participants with high subsequent knowledge (p = 0.046). Conclusions: An educational intervention on donation of organs and tissues for transplant purposes is an effective strategy to increase and reinforce the knowledge of the general population.

Introducción: la falta de información relacionada con la donación de órganos y tejidos resulta devastadora para aquellos pacientes en necesidad de un trasplante, y requiere de una solución basada en el cambio de percepción social mediante intervenciones educativas. Objetivo: determinar el nivel de conocimiento de la población general posterior a una intervención educativa sobre la donación de órganos y tejidos en el Hospital de Cardiología No. 34. Métodos: estudio de intervención educativa con medición antes y después, prospectivo. Instrumento validado mediante fórmula de Kuder-Richardson con coeficiente de fiabilidad de 0.74. La población de estudio se conformó por la población general en las salas de espera de la UMAE No. 34. Las asociaciones con valores de p < 0.05 se consideraron estadísticamente significativas. Resultados: se aplicaron 266 instrumentos de evaluación en 133 participantes. La intervención educativa contribuyó a aumentar el nivel de conocimiento (p = 0.0001). El nivel de conocimiento posterior a la intervención fue mayor en los participantes jóvenes (p = 0.013) y en aquellos con estudios universitarios (p = 0.0001). La relación entre la edad y el nivel de conocimiento posterior mostró significancia favorable en la intención de donación en jóvenes con conocimiento posterior alto (p = 0.046). Conclusiones: una intervención educativa sobre la donación de órganos y tejidos con fines de trasplantes es una estrategia eficaz para aumentar y reforzar el conocimiento de la población general.

Identification of variants in genes associated with hypertrophic cardiomyopathy in Mexican patients.

The objective of this work was to identify genetic variants in Mexican patients diagnosed with hypertrophic cardiomyopathy (HCM). According to world literature, the genes mainly involved are MHY7 and MYBPC3, although variants have been found in more than 50 genes related to heart disease and sudden death, and to our knowledge there are no studies in the Mexican population. These variants are reported and classified in the ClinVar (PubMed) database and only some of them are recognized in the Online Mendelian Information in Men (OMIM). The present study included 37 patients, with 14 sporadic cases and 6 familial cases, with a total of 21 index cases. Next-generation sequencing was performed on a predesigned panel of 168 genes associated with heart disease and sudden death. The sequencing analysis revealed twelve (57%) pathogenic or probably pathogenic variants, 9 of them were familial cases, managing to identify pathogenic variants in relatives without symptoms of the disease. At the molecular level, nine of the 12 variants (75%) were single nucleotide changes, 2 (17%) deletions, and 1 (8%) splice site alteration. The genes involved were MYH7 (25%), MYBPC3 (25%) and ACADVL, KCNE1, TNNI3, TPM1, SLC22A5, TNNT2 (8%). In conclusion; we found five variants that were not previously reported in public databases. It is important to follow up on the reclassification of variants, especially those of uncertain significance in patients with symptoms of the condition. All patients included in the study and their relatives received family genetic counseling.

[Donation in the Cardiology Hospital No. 34]. / Donación en el Hospital de Cardiología No. 34.

BACKGROUND: Chronic degenerative diseases have become a challenge for the Mexican health system. Faced with this problem, health institutions have focused on the therapeutic potential of organ and tissue donation and transplantation. OBJECTIVE: To analyze the experience of the donation program and to identify areas of opportunity at the Hospital de Cardiología No. 34 (Hospital of Cardiology Number 34), in Monterrey, Nuevo León, Mexico. MATERIAL AND METHODS: Observational, cross-sectional, and retrospective study. The study population was made up by deaths and successful interviews. Only groupings with values of p < 0.05 were considered statistically significant. RESULTS: A global of 1947 deaths were registered and a total of 210 interviews were carried out; 83 (39.5%) family members accepted to donate and 127 (60.5%) refused to donate. Only 3 associations between variables had significant statistical value. The year was an important determinant in the increase in effective donations (p = 0.010), and so was the month of the year (p = 0.037), obtaining more positive interviews in the second half of the year; finally, the shift also contributed to the family response (p = 0.012), with the morning shift being the best shift to conduct a successful family interview. CONCLUSIONS: It is imperative to conduct studies that analyze and describe the experience of donation programs to promote and encourage the value of donation.

INTRODUCCIÓN: las enfermedades crónico-degenerativas se han convertido en un desafío para el sistema de salud mexicano. Frente a este problema, las instituciones sanitarias se han enfocado en el potencial terapéutico de la donación y el trasplante de órganos y tejidos. OBJETIVO: analizar la experiencia del programa de donación e identificar áreas de oportunidad en el Hospital de Cardiología No. 34, en Monterrey, Nuevo León, México. MATERIAL Y MÉTODOS: estudio observacional, transversal y retrospectivo. La población de estudio se conformó por defunciones y entrevistas exitosas. Únicamente agrupaciones con valores de p < 0.05 se consideraron estadísticamente significativas. RESULTADOS: se registró un global de 1947 defunciones y se efectuaron en total 210 entrevistas; 83 (39.5%) disponentes secundarios aceptaron donar y 127 (60.5%) se negaron. Solo tres asociaciones entre variables tuvieron valor estadístico significativo. El año fue un determinante importante en el incremento de las donaciones efectivas (p = 0.010) y también lo fue el mes del año (p = 0.037), pues se obtuvieron más entrevistas positivas en el segundo semestre del año; finalmente, el turno también contribuyó en la respuesta familiar (p = 0.012) y fue el turno matutino el mejor para hacer una entrevista familiar exitosa. CONCLUSIONES: es imperativo llevar a cabo estudios que analicen y describan la experiencia del programa de donación para promover y fomentar el valor de la donación.

Curcumin Attenuates the Pathogenicity of Entamoeba histolytica by Regulating the Expression of Virulence Factors in an Ex-Vivo Model Infection.

Infection with the enteric protozoan Entamoeba histolytica is still a serious public health problem, especially in developing countries. Amoebic liver abscess (ALA) is the most common extraintestinal manifestation of the amoebiasis, and it can lead to serious and potentially life-threatening complications in some people. ALA can be cured by metronidazole (MTZ); however, because it has poor activity against luminal trophozoites, 40-60% of treated patients get repeated episodes of invasive disease and require repeated treatments that can induce resistance to MTZ, this may emerge as an important public health problem. Anti-virulence strategies that impair the virulence of pathogens are one of the novel approaches to solving the problem. In this study, we found that low doses of curcumin (10 and 50 µM) attenuate the virulence of E. histolytica without affecting trophozoites growth or triggering liver injury. Curcumin (CUR) decreases the expression of genes associated with E. histolytica virulence (gal/galnac lectin, ehcp1, ehcp5, and amoebapore), and is correlated with significantly lower amoebic invasion. In addition, oxidative stress is critically involved in the etiopathology of amoebic liver abscess; our results show no changes in mRNA expression levels of superoxide dismutase (SOD) and catalase (CAT) after E. histolytica infection, with or without CUR. This study provides clear evidence that curcumin could be an anti-virulence agent against E. histolytica, and makes it an attractive potential starting point for effective treatments that reduce downstream amoebic liver abscess.

Development of a Novel Ex-vivo 3D Model to Screen Amoebicidal Activity on Infected Tissue.

Amoebiasis is a parasitic disease that causes thousands of deaths every year, its adverse effects and resistance to conventional treatments have led to the search of new treatment options, as well as the development of novel screening methods. In this work, we implemented a 3D model of intestine and liver slices from hamsters that were infected ex vivo with virulent E. histolytica trophozoites. Results show preserved histology in both uninfected tissues as well as ulcerations, destruction of the epithelial cells, and inflammatory reaction in intestine slices and formation of micro abscesses, and the presence of amoebae in the sinusoidal spaces and in the interior of central veins in liver slices. The three chemically synthetized compounds T-001, T-011, and T-016, which act as amoebicides in vitro, were active in both infected tissues, as they decreased the number of trophozoites, and provoked death by disintegration of the amoeba, similar to metronidazole. However, compound T-011 induced signs of cytotoxicity to liver slices. Our results suggest that ex vivo cultures of precision-cut intestinal and liver slices represent a reliable 3D approach to evaluate novel amoebicidal compounds, and to simultaneously detect their toxicity, while reducing the number of experimental animals commonly required by other model systems.

Breast Organotypic Cancer Models.

Breast cancer is the most common cancer type diagnosed in women, it represents a critical public health problem worldwide, with 1,671,149 estimated new cases and nearly 571,000 related deaths. Research on breast cancer has mainly been conducted using two-dimensional (2D) cell cultures and animal models. The usefulness of these models is reflected in the vast knowledge accumulated over the past decades. However, considering that animal models are three-dimensional (3D) in nature, the validity of the studies using 2D cell cultures has recently been questioned. Although animal models are important in cancer research, ethical questions arise about their use and usefulness as there is no clear predictivity of human disease outcome and they are very expensive and take too much time to obtain results. The poor performance or failure of most cancer drugs suggests that preclinical research on cancer has been based on an over-dependence on inadequate animal models. For these reasons, in the last few years development of alternative models has been prioritized to study human breast cancer behavior, while maintaining a 3D microenvironment, and to reduce the number of experiments conducted in animals. One way to achieve this is using organotypic cultures, which are being more frequently explored in cancer research because they mimic tissue architecture in vivo. These characteristics make organotypic cultures a valuable tool in cancer research as an alternative to replace animal models and for predicting risk assessment in humans. This chapter describes the cultures of multicellular spheroids, organoids, 3D bioreactors, and tumor slices, which are the most widely used organotypic models in breast cancer research.

[Myocardial infarction in young mexicans associated to metabolic syndrome]. / Infarto agudo al miocardio en jóvenes mexicanos asociado a síndrome metabólico.

BACKGROUND: Acute coronary diseases are catastrophic, especially in young patients. OBJECTIVE: To determine the risk of metabolic syndrome (MS) for premature acute myocardial infarction (AMI), combined with familial, behavioral, and nutritional factors in the northeast of Mexico. MATERIAL AND METHODS: This is a case control study of patients less than 47 years of age with no personal history of angina, AMI, or cerebrovascular disease. Cases corresponded to patients with AMI (incident and primary cases; n = 55) and controls were blood donors located at the same hospital (n = 55). Behavioral, nutritional, and cardiometabolic risk factors were measured. Multivariate logistic regression was used for estimating odds ratios (OR) and 95% confidence intervals (95% CI). RESULTS: MS increased the risk for premature AMI (95% CI: 1.73-39.5) eightfold, followed by smoking (OR: 7.76; 95% CI: 1.27-47.3), family history of AMI or sudden death (OR: 11.0; 95% CI: 2.03-60.4), and sedentary lifestyle (OR: 2.26; 95% CI: 2.52-9.80), independent of potential confounders. CONCLUSIONS: The study highlights the magnitude of the risk of MS for AMI in Mexican young adults. The phenomenon of coronary diseases among young adults needs essential attention from the health sector.

Organotypic culture of breast tumor explants as a multicellular system for the screening of natural compounds with antineoplastic potential.

Breast cancer is the leading cause of death in women worldwide. The search for novel compounds with antitumor activity, with less adverse effects and higher efficacy, and the development of methods to evaluate their toxicity is an area of intense research. In this study we implemented the preparation and culture of breast tumor explants, which were obtained from precision-cut breast tumor slices. In order to validate the model we are proposing to screen antineoplastic effect of natural compounds, we selected caffeic acid, ursolic acid, and rosmarinic acid. Using the Krumdieck tissue slicer, precision-cut tissue slices were prepared from breast cancer samples; from these slices, 4 mm explants were obtained and incubated with the selected compounds. Viability was assessed by Alamar Blue assay, LDH release, and histopathological criteria. Results showed that the viability of the explants cultured in the presence of paclitaxel (positive control) decreased significantly (P < 0.05); however, tumor samples responded differently to each compound. When the explants were coincubated with paclitaxel and compounds, a synergic effect was observed. This study shows that ex vivo culture of breast cancer explants offers a suitable alternative model for evaluating natural or synthetic compounds with antitumor properties within the complex microenvironment of the tumor.

Induction of virulence factors, apoptosis, and cytokines in precision-cut hamster liver slices infected with Entamoeba histolytica.

Precision-cut liver slices (PCLS) are mainly used to evaluate hepatotoxicity and metabolism of chemicals, as well as to study mechanisms of liver damage and repair. However, recently they have been used as a system to study amoebic infections. The aim of this study was to validate this model as an alternative for experimental amoebic liver absess (ALA) in animals. To do this, the PCLS was analyzed for the expression of amoebapore and cysteine proteinases 1 and 5, three of the most studied virulence factors of Entamoeba histolytica, as well as the induction of apoptosis and cytokines production in response to the ex vivo infection. PCHLS were prepared with the Brendel-Vitron tissue slicer and then, infected with 200,000 trophozoites of E. histolytica. Samples were taken at 0, 6, 12, 18, and 24 h and compared to control non-infected slices. Morphological studies were performed in order to verify the infection; while apoptosis was studied by TUNEL and PAS techniques. The expression of cysteine proteinases (1 and 5), and amoebapore, was analyzed by real-time PCR. By using ELISA assays, the production of cytokines was also studied. PCHLS were found to be a reproducible infection system, and E. histolytica caused the expression of cysteine proteinases and amoebapore in infected slices. At the same time, trophozoites induce release of cytokines and apoptotic death of the hepatocytes close to them. PCHLS represent a new and suitable alternative model to study the pathogenesis of hepatic amoebiasis.

Precision-cut hamster liver slices as an ex vivo model to study amoebic liver abscess.

Entamoeba histolytica is the etiological agent of amoebiasis, the second cause of global morbidity and mortality due to parasitic diseases in humans. In approximately 1% of the cases, amoebas penetrate the intestinal mucosa and spread to other organs, producing extra-intestinal lesions, among which amoebic liver abscess (ALA) is the most common. To study ALA, in vivo and in vitro models are used. However, animal models may pose ethical issues, and are time-consuming and costly; and cell cultures represent isolated cellular lineages. The present study reports the infection of precision-cut hamster liver slices with Entamoeba histolytica trophozoites. The infection time-course, including tissue damage, parallels findings previously reported in the animal model. At the same time amoebic virulence factors were detected in the infected slices. This new model to study ALA is simple and reproducible, and employs less than 1/3 of the hamsters required for in vivo analyses.