The Caenorhabditis elegans protein SOC-3 permits an alternative mode of signal transduction by the EGL-15 FGF receptor.

Fibroblast Growth Factors and their receptors (FGFRs) comprise a cell signaling module that can stimulate signaling by Ras and the kinases Raf, MEK, and ERK to regulate animal development and homeostatic functions. In Caenorhabditis elegans, the sole FGFR ortholog EGL-15 acts with the GRB2 ortholog SEM-5 to promote chemoattraction and migration by the sex myoblasts (SMs) and fluid homeostasis by the hypodermis (Hyp7). Cell-specific differences in EGL-15 signaling were suggested by the phenotypes caused by egl-15(n1457), an allele that removes a region of its C-terminal domain (CTD) known to bind SEM-5. To determine how mutations altered EGL-15 activity in the SMs and Hyp7, we used the kinase reporter ERK-KTR to measure activation of the ERK ortholog MPK-1. Consequences of egl-15(n1457) were cell-specific, resulting in loss of MPK-1 activity in the SMs and elevated activity in Hyp7. Previous studies of Hyp7 showed that loss of the CLR-1 phosphatase causes a fluid homeostasis defect termed "Clear" that is suppressed by reduction of EGL-15 signaling, a phenotype termed "Suppressor of Clear" (Soc). To identify mechanisms that permit EGL-15 signaling in Hyp7, we conducted a genetic screen for Soc mutants in the clr-1; egl-15(n1457) genotype. We report the identification of SOC-3, a protein with putative SEM-5-binding motifs and PH and PTB domains similar to DOK and IRS proteins. In combination with the egl-15(n1457) mutation, loss of either soc-3, the GAB1 ortholog soc-1, or the SHP2 ortholog ptp-2, reduced MPK-1 activation. We generated alleles of soc-3 to test the requirement for the SEM-5-binding motifs, finding that residue Tyr356 is required for function. We propose that EGL-15-mediated SM chemoattraction relies solely on the direct interaction between SEM-5 and the EGL-15 CTD. In Hyp7, EGL-15 signaling uses two mechanisms: the direct SEM-5 binding mechanism; and an alternative, CTD-independent mechanism involving SOC-3, SOC-1, and PTP-2. This work demonstrates that FGF signaling uses distinct, tissue-specific mechanisms in development, and identifies SOC-3 as a potential adaptor that facilitates Ras pathway activation by FGFR.

Novel Antennapedia and Ultrabithorax trimeric complexes with TBP and Exd regulate transcription.

BACKGROUND: Hox proteins interact with DNA and many other proteins, co-factors, transcriptional factors, chromatin remodeling components, non-coding RNAs and even the extracellular matrix that assembles the Hox complexes. The number of interacting partners continues to grow with diverse components and more transcriptional factors than initially thought. Hox complexes present many activities, but their molecular mechanisms to modulate their target genes remain unsolved. RESULTS: In this paper we showed the protein-protein interaction of Antp with Ubx through the homeodomain using BiFC in Drosophila. Analysis of Antp-deletional mutants showed that AntpHD helixes 1 and 2 are required for the interaction with Ubx. Also, we found a novel interaction of Ubx with TBP, in which the PolyQ domain of TBP is required for the interaction. Moreover, we also detected the formation of two new trimeric complexes of Antp with Ubx, TBP and Exd using BiFC-FRET; these proteins, however, do not form a trimeric interaction with BIP2 or TFIIEß. The novel trimeric complexes reduced Antp transcriptional activity, indicating that they could confer specificity for repression. CONCLUSIONS: Our results increase the number of transcriptional factors in the Antp and Ubx interactomes that form two novel trimeric complexes with TBP and Exd. We also report a new Ubx interaction with TBP. These novel interactions provide important clues of the dynamics of Hox-interacting complexes involved in transcriptional regulation, contributing to better understand Hox function.

T-FINDER: A highly sensitive, pan-HLA platform for functional T cell receptor and ligand discovery.

Effective, unbiased, high-throughput methods to functionally identify both class II and class I HLA-presented T cell epitopes and their cognate T cell receptors (TCRs) are essential for and prerequisite to diagnostic and therapeutic applications, yet remain underdeveloped. Here, we present T-FINDER [T cell Functional Identification and (Neo)-antigen Discovery of Epitopes and Receptors], a system to rapidly deconvolute CD4 and CD8 TCRs and targets physiologically processed and presented by an individual's unmanipulated, complete human leukocyte antigen (HLA) haplotype. Combining a highly sensitive TCR signaling reporter with an antigen processing system to overcome previously undescribed limitations to target expression, T-FINDER both robustly identifies unknown peptide:HLA ligands from antigen libraries and rapidly screens and functionally validates the specificity of large TCR libraries against known or predicted targets. To demonstrate its capabilities, we apply the platform to multiple TCR-based applications, including diffuse midline glioma, celiac disease, and rheumatoid arthritis, providing unique biological insights and showcasing T-FINDER's potency and versatility.

Genotyping of Toxoplasma gondii in domestic animals from Campeche, México, reveals virulent genotypes and a recombinant ROP5 allele.

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.

Silica-coated magnetic particles for efficient RNA extraction for SARS-CoV-2 detection.

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.

Efficient catalytic activity of NiO and CeO2 films in benzoic acid removal using ozone.

This research focuses on the synthesis of NiO and CeO2 thin films using spray pyrolysis for the removal of benzoic acid using ozone as an oxidant. The results indicate that the addition of CeO2 films significantly enhances the mineralization of benzoic acid, achieving a rate of over 80% as the CeO2 films react with ozone to produce strong oxidant species, such as hydroxyl radicals, superoxide radicals, and singlet oxygen as demonstrated by the presence of quenchers in the reaction system. The difference in catalytic activity between NiO and CeO2 films was analyzed via XPS technique; specifically, hydroxyl oxygen groups in the CeO2 film were greater in number than those in the NiO film, thus benefitting catalytic oxidation as these species are considered active oxidation sites. The effects of nozzle-substrate distances and deposition time during the synthesis of the films on benzoic acid removal efficiency were also explored. Based on XRD characterization, it was established that the NiO and CeO2 films were polycrystalline with a cubic structure. NiO spherical nanoparticles were well-distributed on the substrate surface, while some pin holes and overgrown clusters were observed in the CeO2 films according to the SEM results. The stability of the CeO2 films after five consecutive cycles confirms their reusability. The retrieval of films is easy because it does not require additional separation methods, unlike the catalyst in powder form. The obtained results indicate that the CeO2 films have potential application in pollutant removal from water through catalytic ozonation.

PhyloString: A web server designed to identify, visualize, and evaluate functional relationships between orthologous protein groups across different phylogenetic lineages.

Proteins are biological units whose essence is defined by their functional relationships with other proteins or biomolecules such as RNA, DNA, lipids, or carbohydrates. These functions encompass enzymatic, structural, regulatory, or physical interaction roles. The STRING database (Nucleic Acids Research, 8 Jan 2021;49(D1): D605-12) provides an index that defines the functional interaction networks between proteins in model organisms. To facilitate the identification, visualization, and evaluation of potential functional networks across organisms from different phylogenetic lineages, we have developed PhyloString (https://biocomputo.ibt.unam.mx/phylostring/), a web server that utilizes the indices of the STRING database. PhyloString decomposes these functional networks into modules, representing cohesive units of proteins grouped based on their similarity of STRING values and the phylogenetic origins of their respective organisms. This study presents and thoroughly discusses examples of such functional networks and their modules identified using PhyloString.

Decisiones por sustitución en personas mayores con deterioro cognitivo y su delegación de autonomía ¿representada? / Surrogate decision-making in older people with cognitive impairment and the delegation of their autonomy: is it represented? / As decisões de substituição em idosos com deficiência cognitiva e sua delegação de autonomia ¿representada?

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.

Non-Targeted Metabolomic Analysis of Arabidopsis thaliana (L.) Heynh: Metabolic Adaptive Responses to Stress Caused by N Starvation.

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.

The E3/E4 ubiquitin ligase UFD-2 suppresses normal and oncogenic signaling mediated by a Raf ortholog in Caenorhabditis elegans.

Signaling by the kinase cascade composed of Raf, MEK, and ERK is critical for animal development and is often inappropriately activated in human malignancies. We sought to identify factors that control signaling mediated by the Caenorhabditis elegans Raf ortholog LIN-45. A genetic screen showed that the degradation of LIN-45 required the E3/E4 ubiquitin ligase UFD-2. Both UFD-2 and its partner, the ATP-dependent segregase CDC-48, were required for the developmental regulation of LIN-45 protein abundance. We showed that UFD-2 acted in the same pathway as the E3 ubiquitin ligase SCFSEL-10 to decrease LIN-45 abundance in cells in which Raf-MEK-ERK signaling was most highly active. UFD-2 also reduced the protein abundance of activated LIN-45 carrying a mutation equivalent to the cancer-associated BRAF(V600E) variant. Our structure-function studies showed that the disruption of LIN-45 domains that mediate protein-protein interactions, including the conserved cysteine-rich domain and 14-3-3 binding motifs, were required for UFD-2-independent degradation of LIN-45. We propose a model in which UFD-2 and CDC-48 act downstream of SCFSEL-10 to remove LIN-45 from its protein interaction partners and facilitate proteasomal targeting and degradation. These findings imply that UFD-2 and CDC-48 may be important for Raf degradation during normal and oncogenic Ras and MAPK signaling in mammalian cells.

Genotyping of toxoplasma gondii isolates from México reveals non-archetypal and potentially virulent strains for mice.

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.

Cytotoxicity Induced by Newly Synthesized Palladium (II) Complexes Lead to the Death of MCF-7 and MDA-MB-435 Cancer Cell Lines.

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.

In Vitro Gastrointestinal Digestion Affects the Bioaccessibility of Bioactive Compounds in Hibiscus sabdariffa Beverages.

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.

COVID-19 Vaccine Equity and Access: Case Study for Health Care Chatbots.

BACKGROUND: Disparities in COVID-19 information and vaccine access have emerged during the pandemic. Individuals from historically excluded communities (eg, Black and Latin American) experience disproportionately negative health outcomes related to COVID-19. Community gaps in COVID-19 education, social, and health care services (including vaccines) should be prioritized as a critical effort to end the pandemic. Misinformation created by the politicization of COVID-19 and related public health measures has magnified the pandemic's challenges, including access to health care, vaccination and testing efforts, as well as personal protective equipment. Information and Communication Technology (ICT) has been demonstrated to reduce the gaps of marginalization in education and access among communities. Chatbots are an increasingly present example of ICTs, particularly in health care and in relation to the COVID-19 pandemic. OBJECTIVE: This project aimed to (1) follow an inclusive and theoretically driven design process to develop and test a COVID-19 information ICT bilingual (English and Spanish) chatbot tool named "Ana" and (2) characterize and evaluate user experiences of these innovative technologies. METHODS: Ana was developed following a multitheoretical framework, and the project team was comprised of public health experts, behavioral scientists, community members, and medical team. A total of 7 iterations of ß chatbots were tested, and a total of 22 ß testers participated in this process. Content was curated primarily to provide users with factual answers to common questions about COVID-19. To ensure relevance of the content, topics were driven by community concerns and questions, as ascertained through research. Ana's repository of educational content was based on national and international organizations as well as interdisciplinary experts. In the context of this development and pilot project, we identified an evaluation framework to explore reach, engagement, and satisfaction. RESULTS: A total of 626 community members used Ana from August 2021 to March 2022. Among those participants, 346 used the English version, with an average of 43 users per month; and 280 participants used the Spanish version, with an average of 40 users monthly. Across all users, 63.87% (n=221) of English users and 22.14% (n=62) of Spanish users returned to use Ana at least once; 18.49% (n=64) among the English version users and 18.57% (n=52) among the Spanish version users reported their ranking. Positive ranking comprised the "smiley" and "loved" emojis, and negative ranking comprised the "neutral," "sad," and "mad" emojis. When comparing negative and positive experiences, the latter was higher across Ana's platforms (English: n=41, 64.06%; Spanish: n=41, 77.35%) versus the former (English: n=23, 35.93%; Spanish: n=12, 22.64%). CONCLUSIONS: This pilot project demonstrated the feasibility and capacity of an innovative ICT to share COVID-19 information within diverse communities. Creating a chatbot like Ana with bilingual content contributed to an equitable approach to address the lack of accessible COVID-19-related information.

Oxidative Damage by 3-nitrotyrosine in Young Adults with Obesity: Its Implication in Chronic and Contagious Diseases.

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.

Úlceras de pie diabético y plasma rico en plaquetas / Healing of diabetic foot ulcers with platelet-rich plasma

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.

The use of ecological analytical tools as an unconventional approach for untargeted metabolomics data analysis: the case of Cecropia obtusifolia and its adaptive responses to nitrate starvation.

Plant metabolomics studies haves revealed new bioactive compounds. However, like other omics disciplines, the generated data are not fully exploited, mainly because the commonly performed analyses focus on elucidating the presence/absence of distinctive metabolites (and/or their precursors) and not on providing a holistic view of metabolomic changes and their participation in organismal adaptation to biotic and abiotic stress conditions. Therefore, spectral libraries generated from Cecropia obtusifolia cell suspension cultures in a previous study were considered as a case study and were reanalyzed herein. These libraries were obtained from a time-course experiment under nitrate starvation conditions using both electrospray ionization modes. The applied methodology included the use of ecological analytical tools in a systematic four-step process, including a population analysis of metabolite α diversity, richness, and evenness (i); a chemometrics analysis to identify discriminant groups (ii); differential metabolic marker identification (iii); and enrichment analyses and annotation of active metabolic pathways enriched by differential metabolites (iv). Our species α diversity results referring to the diversity of metabolites represented by mass-to-charge ratio (m/z) values detected at a specific retention time (rt) (an uncommon way to analyze untargeted metabolomic data) suggest that the metabolome is dynamic and is modulated by abiotic stress. A total of 147 and 371 m/z_rt pairs was identified as differential markers responsive to nitrate starvation in ESI- and ESI+ modes, respectively. Subsequent enrichment analysis showed a high degree of completeness of biosynthetic pathways such as those of brassinosteroids, flavonoids, and phenylpropanoids.

Transcranial electric stimulation motor evoked potentials for cervical spine intraoperative monitoring complications: systematic review and illustrative case of cardiac arrest.

PURPOSE: We show a systematic review of known complications during intraoperative neuromonitoring (IONM) using transcranial electric stimulation motor evoked potentials (TES-MEP) on cervical spine surgery, which provides a summary of the main findings. A rare complication during this procedure, cardiac arrest by cardioinhibitory reflex, is also described. METHODS: Findings of 523 scientific papers published from 1995 onwards were reviewed in the following databases: CENTRAL, Cochrane Library, Embase, Google Scholar, Ovid, LILACS, PubMed, and Web of Science. This study evaluated only complications on cervical spine surgery undergoing TES-MEP IONM. RESULTS: The review of the literature yielded 13 studies on the complications of TES-MEP IONM, from which three were excluded. Five studies are case series; the rest are case reports. Overall, 169 complications on 167 patients were reported in a total of 38,915 patients, a global prevalence of 0.43%. The most common complication was tongue-bite in 129 cases, (76.3% of all complication events). Tongue-bite had a prevalence of 0.33% (CI 95%, 0.28-0.39%) in all patients on TES-MEP IONM. A relatively low prevalence of severe complications was found: cardiac-arrhythmia, bradycardia and seizure, the prevalence of this complications represents only one case in all the sample. Alongside, we report the occurrence of cardiac arrest attributable to TES-MEP IONM. CONCLUSIONS: This systematic review shows that TES-MEP is a safe procedure with a very low prevalence of complications. To our best knowledge, asystole is reported for the first time as a complication during TES-MEP IONM.

Acute Disseminated Toxoplasmosis in Two Specimens of Macropus rufogriseus Caused by a Genotype so far Exclusive to South America.

Macropods are included among the species considered highly susceptible to Toxoplasma gondii infection. Clinically, it is difficult to distinguish between acute toxoplasmosis due to primary infection and reactivation of chronic latent infection in susceptible species until pathologic studies are performed. Here, we described the clinical cases and lesions found in two deceased Bennett's wallabies (Macropus rufogriseus) with a presumptive diagnosis of toxoplasmosis, as well as the genetic characterization of the T. gondii isolates obtained from these specimens. Both animals presented acute infection lesions in the lungs, liver, spleen and lymph nodes associated to T. gondii infection. Histopathology and immunohistochemistry also demonstrated tissue cysts of different sizes, indicating that the wallabies were previously infected with this parasite. Two isolates were obtained, one from each specimen and the molecular characterization was done; both isolates were the ToxoDB #116 genotype. This is the first study that reports the isolation of this particular genotype outside South America, and given the histopathological findings, it could be considered virulent for this species. The dynamics of infection that T. gondii is causing in definitive and intermediate hosts in a region allows us to know the risks to which the animals and humans that live in the area are exposed, and in the future to implement a preventive medicine plan against this parasite.