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Toxoplasma gondii has at least 318 genotypes distributed worldwide, and tropical regions usually have greater genetic diversity. Campeche is a state located in the southeastern region of México and has favourable climate conditions for the replication and dissemination of this protozoan, similar to those in South American countries where broad genetic diversity has been described. Thus, in this study, 4 T. gondii isolates were obtained from tissues of stray dogs and free-range chickens in Campeche, México, and were genotyped by Mn-PCR-RFLP with 10 typing markers (SAG1, altSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) and 5 virulence markers (CS3, ROP16, ROP17, ROP18 and ROP5) to provide new information about the distribution and virulence prediction of T. gondii genotypes. Two isolates of T. gondii genotype #116 and 2 of genotype #38 were obtained from stray dogs and chickens, respectively. The parasite load found in these species was between <50 and more than 35 000 tachyzoites per mg of tissue. Virulence marker genotyping revealed a recombinant 1&3 ROP5 RFLP pattern in 2 ToxoDB #116 isolates with no prediction of virulence in a murine model, while in the 2 ToxoDB #38 isolates, the ROP18/ROP5 combination predicted high virulence. Considering all the typed markers, there is a predominance of type I and III alleles, as constantly reported for the isolates characterized in various regions of México. It is crucial to determine their phenotype to corroborate the genetic virulence profile of the T. gondii isolates obtained in this study.

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Molecular diagnostic methods to detect and quantify viral RNA in clinical samples rely on the purification of the genetic material prior to reverse transcription polymerase chain reaction (qRT-PCR). Due to the large number of samples processed in clinical laboratories, automation has become a necessity in order to increase method processivity and maximize throughput per unit of time. An attractive option for isolating viral RNA is based on the magnetic solid phase separation procedure (MSPS) using magnetic microparticles. This method offers the advantage over other alternative methods of making it possible to automate the process. In this study, we report the results of the MSPS method based on magnetic microparticles obtained by a simple synthesis process, to purify RNA from oro- and nasopharyngeal swab samples of patients suspected of COVID-19 provided by three diagnostic laboratories located in the Buenos Aires Province, Argentina. Magnetite nanoparticles of Fe3O4 (MNPs) were synthesized by the coprecipitation method and then coated with silica (SiO2) produced by hydrolysis of tetraethyl orthosilicate (TEOS). After preliminary tests on samples from the A549 human lung cell line and swabs, an extraction protocol was developed. The quantity and purity of the RNA obtained were determined by gel electrophoresis, spectrophotometry, and qRT-PCR. Tests on samples from naso- and oropharyngeal swabs were performed in order to validate the method for RNA purification in high-throughput SARS-CoV-2 diagnosis by qRT-PCR. The method was compared to the spin columns method and the automated method using commercial magnetic particles. The results show that the method developed is efficient for RNA extraction from nasal and oropharyngeal swab samples, and also comparable to other extraction methods in terms of sensitivity for SARS-CoV-2 detection. Of note, this procedure and reagents developed locally were intended to overcome the shortage of imported diagnostic supplies as the sudden spread of COVID-19 required unexpected quantities of nucleic acid isolation and diagnostic kits worldwide.

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Objetivo: Analizar las consideraciones éticas que surgen en situaciones donde las personas mayores con deterioro cognitivo delegan sus decisiones a terceros. Desarrollo: Trabajo de reflexión basado en una revisión narrativa desde diferentes fuentes de información. El envejecimiento de la población es un fenómeno donde se evidencian múltiples desafíos a atender. Chile se encuentra en una etapa de envejecimiento avanzado estimando una esperanza de vida sobre los 85 años para el 2050. La Organización Mundial de la Salud promueve el envejecimiento saludable mediante el fortalecimiento de la capacidad funcional como indicador de bienestar. Existen ocasiones donde las personas mayores delegan a un tercero su capacidad de decidir. Las decisiones por sustitución, que abordan dimensiones éticas y legales deben proteger la dignidad de la persona en todo momento, independientemente de su condición de salud y representar plenamente el derecho de autonomía cedido. Conclusiones. La discusión sobre las decisiones por sustitución y la protección de la autonomía, incluso en situaciones de limitaciones cognitivas, subraya la necesidad de tratar a las personas mayores con respeto y ética, desafiando los estereotipos negativos y evitando conductas discriminatorias y estigmatizantes. La importancia de promover la igualdad, inclusión, información y el respeto de los derechos de las personas mayores tiene un impacto directo en el bienestar y calidad de vida, elemento fundamental que la salud pública promueve bajo el contexto del envejecimiento activo, saludable y digno.

Objective: To analyze the ethical considerations in situations where older people with cognitive impairment delegate their decisions to third parties. Development: Reflective practice based on a narrative review of different sources of information. Population aging is a phenomenon where multiple challenges need to be addressed. Chile is at an advanced stage of population aging, with an estimated life expectancy of over 85 years to be reached by 2050. The World Health Organization (WHO) promotes healthy aging by strengthening functional capacity as an indicator of well-being. There are circumstances in which older people delegate their decision-making capacity to a third party. Surrogate decision-making, with its ethical and legal dimensions, should protect the person's dignity at all times, regardless of the state of their health, and fully represent their ceded right of autonomy. Conclusions: The discussion on surrogate decision-making and the protection of autonomy, even in the context of cognitive impairment, highlights the need to treat older people ethically and respectfully, challenging negative stereotypes and avoiding discriminatory and stigmatizing behaviors. Promoting equality, inclusion, information, and respect for the rights of older adults has a direct impact on their well-being and quality of life, which are fundamental elements that public health promotes in the context of active, healthy, and dignified aging.

Objetivo: Analisar as considerações éticas que surgem em situações em que pessoas idosas com deficiência cognitiva delegam as suas decisões a terceiros. Desenvolvimento: Trabalho de reflexão baseado numa revisão narrativa a partir de diferentes fontes de informação. O envelhecimento da população é um fenómeno onde existem múltiplos desafios a enfrentar. O Chile encontra-se numa fase avançada de envelhecimento, estimando uma esperança de vida superior a 85 anos até 2050. A Organização Mundial de Saúde promove o envelhecimento saudável através do fortalecimento da capacidade funcional como indicador de bem-estar. Há ocasiões em que os idosos delegam a sua capacidade de decisão a terceiros. As decisões de substituição, que abordam as dimensões éticas e legais, devem proteger a dignidade da pessoa em todos os momentos, independentemente do seu estado de saúde, e representar plenamente o direito concedido à autonomia. Conclusões: A discussão sobre as decisões de substituição e a proteção da autonomia, mesmo em situações de limitações cognitivas, sublinha a necessidade de tratar os idosos com respeito e ética, desafiando estereótipos negativos e evitando comportamentos discriminatórios e estigmatizantes. A importância da promoção da igualdade, da inclusão, da informação e do respeito pelos direitos dos idosos tem impacto direto no bem-estar e na qualidade de vida, elemento fundamental que a saúde pública promove no contexto do envelhecimento ativo, saudável e digno.

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As sessile organisms, plants develop the ability to respond and survive in changing environments. Such adaptive responses maximize phenotypic and metabolic fitness, allowing plants to adjust their growth and development. In this study, we analyzed the metabolic plasticity of Arabidopsis thaliana in response to nitrate deprivation by untargeted metabolomic analysis and using wild-type (WT) genotypes and the loss-of-function nia1/nia2 double mutant. Secondary metabolites were identified using seedlings grown on a hydroponic system supplemented with optimal or limiting concentrations of N (4 or 0.2 mM, respectively) and harvested at 15 and 30 days of age. Then, spectral libraries generated from shoots and roots in both ionization modes (ESI +/-) were compared. Totals of 3407 and 4521 spectral signals (m/z_rt) were obtained in the ESI+ and ESI- modes, respectively. Of these, approximately 50 and 65% were identified as differentially synthetized/accumulated. This led to the presumptive identification of 735 KEGG codes (metabolites) belonging to 79 metabolic pathways. The metabolic responses in the shoots and roots of WT genotypes at 4 mM of N favor the synthesis/accumulation of metabolites strongly related to growth. In contrast, for the nia1/nia2 double mutant (similar as the WT genotype at 0.2 mM N), metabolites identified as differentially synthetized/accumulated help cope with stress, regulating oxidative stress and preventing programmed cell death, meaning that metabolic responses under N starvation compromise growth to prioritize a defensive response.

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Genotyping and virulence studies of Toxoplasma gondii are essential to investigate the pathogenesis of strains circulating worldwide. In this study, eight T. gondii isolates obtained from a congenitally infected newborn, a calf, two cats, three dogs, and a wallaby from five states of México were genotyped by Mn-PCR-RFLP with 11 typing markers (SAG1, SAG2 5'3', alt. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico), five virulence markers (CS3, ROP16, ROP17, ROP18 and ROP5), 15 microsatellite markers (TUB-2, W35, TgM-A, B18, B17, M33, IV.1, XI.1, M48, M102, N60, N82, AA, N61, N83), and sequencing. A phylogenetic network was built to determine the relationship between Mexican isolates and those reported worldwide. Six different genotypes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), ToxoDB #8, #10, #28 (n = 3), #48, #116, and #282. Genotyping by microsatellite analysis differentiated the three PCR-RFLP genotype #28 isolates into two strains, revealing a total of seven microsatellite genotypes. Three different allele combinations of ROP18/ROP5 virulence markers were also found, 3/3, 1/1, and 4/1. The last two combinations are predicted to be highly virulent in the murine model. According to the phylogenetic network, the T. gondii strains studied here are related to archetypal strains I and III, but none are related to the strains previously reported in México. The genotypes identified in this study in different species of animals demonstrate the great genetic diversity of T. gondii in México. The ToxoDB-PCR-RFLP #28 genotype was found in three isolates from different hosts and states. Additionally, four of the isolates are predicted to be highly virulent in mice. The next step will be to perform in vitro and in vivo assays to determine the phenotype of these T. gondii isolates in murine models.

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Purpose: Breast cancer is the most common female malignancy and melanoma is the most lethal type of skin cancer. Traditional therapy for cancer treatment is far from satisfactory due to drug resistance and side effects, thus a search for new medicines is being emphasized. Palladium(II) complexes have been reported as anticancer potential agents. In this work, the anticancer activities and cell death induction of a new series of square-planar Pd(II) complexes were evaluated against MCF-7 and MDA-MB-435 cancer cells. Methods: MCF-7 (breast carcinoma) and MDA-MB-435 (melanoma) cells were cultivated, and treated with ligand and Pd(II) complexes. Cell growth, migration and adhesion inhibition, morphological alterations, cell death induction and, DNA interaction upon treatment were studied. Results: Pd(II) complexes exhibited both short and long-term antiproliferative effects on both cell lines, reducing by 80% cell growth in the SRB assay and abolishing longterm proliferation, estimated by the clonogenic assay. Complexes reduced significantly (P<0.05) cell migration and adhesion when compared to the control group. Complexes induced morphological alterations in cell lines and significant (P<0.05) cellular shrinkage. Cell death was induced and the complexes were able to interact with DNA, inducing cleavage of double-stranded DNA, which may account for the complexes cytotoxic effects, observed against both MCF-7 and MDA-MB-435 cells. Conclusion: Overall, the complexes exhibited cytotoxic activities and induced cell death. These observations emphasize an anticancer role with a potential therapeutic value for Pd(II) complexes to improve the outcome of patients with breast cancer and melanoma.

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Hibiscus sabdariffa possess great versatility to be used as an ingredient for a whole range of products with natural-based ingredients, which are growing in popularity due to the health benefits of bioactive compounds (BC). Therefore, the objective of this study was to characterize the BC content in Hibiscus beverages and to evaluate their in vitro bioaccessibility. Results showed significant differences (p < 0.05) in the total contents of BC prior to the in vitro intestinal digestion. Hibiscus acid was the most abundant compound identified. Thirty-five compounds were identified in the Hibiscus beverage at the initial stage, while a maximum of 15 compounds were quantified in the different fractions of gastrointestinal digestion. After digestion, significant differences were found compared with the initial content of BC. That phenolic acids were the less bioaccessible group, while flavonoids were the most diverse. Principal components analysis showed different clusters and changes in the profiles of BC present at the initial stage and those bioaccessible, showing that intestinal digestion significantly affects the BC profile of the beverage.

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INTRODUCTION: Cellular damage by oxidation occurs in numerous chronic diseases, such as obesity, type II diabetes, cardiovascular disease, nonalcoholic fatty liver, etc. The oxidized compound 3-nitrotyrosine is a marker of oxidative stress and protein oxidation damage. OBJECTIVE: The article aims to assess whether 3-nitrotyrosine levels are higher in young people with obesity than in the same population without obesity. METHODS: Anthropometry and blood chemistry analyses were performed on 24 young Mexican participants (18-30 years old), categorized into two groups based on their waist circumference: Withobesity (≥ 80 cm women; ≥ 90 cm men) and without-obesity (<80 cm women; <90 cm men). Additionally, 3-nitrotyrosine blood values were quantified by ELISA. RESULTS: Except for HDL-cholesterol, the mean values of lipids increased in women and men with obesity (p<0.05), and 3-nitrotyrosine concentration (nM/µg total protein) was higher by 60% in the group with-obesity compared to the group without-obesity, both for women (66.21 ± 23.85 vs. 40.69 ± 16.25, p<0.05) and men (51.72 ± 20.56 vs. 30.52 ± 5.21, p<0.05). CONCLUSION: Oxidative damage measured by compound 3-nitrotyrosine was higher in the group with obesity than in the group without obesity, which, if not controlled, could lead to a chronic oxidative condition and thereby to a degree of cellular aging with adverse health effects.

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Introducción: El plasma rico en plaquetas (PRP) se encuentra en desarrollo desde los años 80, asociado con aplicaciones en medicina cardíaca, traumatológica y dermatológica. El PRP es una preparación autóloga, con una concentración de plaquetas por sobre el valor normal en sangre periférica, que se obtiene a partir de su centrifugación siguiendo diferentes protocolos que fueron valorados en distintas revisiones sistemáticas. El plasma luego se activa con trombina o cloruro cálcico, o por lisis física (ultrasonido o frizado). El PRP contiene factores de crecimiento, citoquinas y proteínas de adhesión que, al aplicarse en la lesión, favorecen la hemostasia, la síntesis de tejido conectivo y la revascularización. El producto se aplica en solución o gel. Se intentaron diferentes clasificaciones del PRP para estandarizarlo, sin éxito. Se conceptualizó la receta del PRP para aplicación clínica, que presentó las siguientes características: valores altos de plaquetas, disminución de la contaminación de glóbulos rojos, presencia de neutrófilos y leucocitos para el éxito terapéutico. Protocolo y casos: Se presenta el protocolo de preparación de PRP de centrifugación única de 7 minutos a 1400 rpm, con activación por medio de gluconato de calcio según la siguiente proporción: 3 ml plasma/0.4 ml gluconato cálcico. Además, se informan tres casos de úlceras de pie diabético de nuestra institución. Conclusión: El PRP no presenta complejidad en su preparación y aplicación, por lo que es factible de realizar en el primer nivel de atención que cuente con los recursos materiales y profesionales con conocimiento en el abordaje de heridas crónicas.

Introduction: Platelet-rich plasma (PRP) has been under development since the 1980s, associated with applications in cardiac, traumatological, and dermatological medicine. PRP is an autologous preparation with a platelet concentration above the normal value in peripheral blood that is obtained from its centrifugation following different protocols that were evaluated in different systematic reviews. The plasma is then activated with thrombin or calcium chloride, or by physical lysis (ultrasound or friz). PRP contains growth factors, cytokines, and adhesion proteins that, when applied to the lesion, favor hemostasis, connective tissue synthesis, and revascularization. The product is applied in solution or gel. Different classifications of the PRP were used with the intention of standardizing the procedure without success. The PRP recipe for clinical application was conceptualized. It presented the following characteristics: high platelet values, decreased red blood cell contamination, presence of neutrophils and leukocytes for therapeutic success. Protocol and cases: The PRP preparation protocol for single centrifugation for 7 minutes at 1400 rpm with activation through calcium gluconate is presented according to the following ratio: 3 ml plasma/0.4 ml of calcium gluconate. Three cases of diabetic foot ulcers from our institution are reported. Conclusion: The PRP does not present complexity in its preparation and application, so it is feasible to perform it in the first level of care that has the material and professional resources with knowledge in the approach to chronic wounds.

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