Secondary metabolites with antimicrobial activity produced by thermophilic bacteria from a high-altitude hydrothermal system.

Thermophilic microorganisms possess several adaptations to thrive in high temperature, which is reflected as biosynthesis of proteins and thermostable molecules, isolation and culture represent a great methodological challenge, therefore High throughput sequencing enables screening of the whole bacterial genome for functional potential, providing rapid and cost-effective information to guide targeted cultures for the identification and characterization of novel natural products. In this study, we isolated two thermophilic bacterial strains corresponding to Bacillus LB7 and Streptomyces LB8, from the microbial mats in the Atacama Desert. By combining genome mining, targeted cultures and biochemical characterization, we aimed to identify their capacity to synthesize bioactive compounds with antimicrobial properties. Additionally, we determined the capability to produce bioactive compounds under controlled in vitro assays and detected by determining their masses by Thin-Layer Chromatography/Mass Spectrometry (TLC/MS). Overall, both isolates can produce antimicrobial (e.g., Myxalamide C by-product) and antioxidants (e.g. Dihydroxymandelic Acid, Amide biotine and Flavone by-products) compounds. Bacillus LB7 strain possesses a more diverse repertoire with 51.95% of total metabolites unmatched, while Streptomyces LB8 favors mainly antioxidants, but has over 70% of unclassified compounds, highlighting the necessity to study and elucidate the structure of novel compounds. Based on these results, we postulate that the uncultured or rare cultured thermophiles inhabiting high-altitude hydrothermal ecosystems in the Atacama Desert offer a promising opportunity to the study of novel microbial bioactive compounds.

Genotypic and morphological identification of opportunistic microorganisms in triple-syringe tubing from dental units.

BACKGROUND: In Mexico and around the world, water in dental units, including triple syringes, comes from municipal chlorinated water mains. The microbial contamination of dental unit water systems constitutes a risk factor for opportunistic infections. OBJECTIVES: The present work aimed to identify the bacteria present in the triple-syringe water lines of dental units at a dental school of a public university in Mexico, with a hypothesis that opportunistic bacteria of importance to human health would be found. MATERIAL AND METHODS: A cross-sectional study was carried-out. A total of 100 samples of triple-syringe tubing from dental units operated by a dental school of a public university in Mexico were analyzed before and after their use in dental practice. Bacterial biofilm was cultured and isolated from the tubing, using standard microbiological methods, and then the species present were identified through 16S rRNA gene sequencing. The characterization of the biofilm was performed by means of scanning electron microscopy (SEM). RESULTS: Bacterial growth was observed in 20% of the non-disinfected and 10% of the disinfected samples, with 11 strains isolated. Six genera and 11 bacterial species were genetically identified. Coagulasenegative staphylococci (CoNS), considered opportunistic human pathogens, were among the most critical microorganisms. Scanning electron microscopy revealed a thick polymeric matrix with multiple bacterial aggregates. CONCLUSIONS: Opportunistic bacteria from human skin and mucous membranes were detected. Under normal conditions, these bacteria are incapable of causing disease, but are potentially harmful to immunosuppressed patients.

Chemical composition and antioxidant properties of native Ecuadorian fruits: Rubus glabratus Kunth, Vaccinium floribundum Kunth, and Opuntia soederstromiana.

The native berries of South America present promising marketing opportunities owing to their high antioxidant content, notably rich in anthocyanin and phenolic compounds. However, Ecuador's endemic fruits, primarily found in the wild, lack comprehensive data regarding their phytochemical composition and antioxidant capacity, underscoring the need for research in this area. Accordingly, this study evaluated the total phenolic, anthocyanin, flavonoid, resveratrol, ascorbic acid, citric acid, sugars, and antioxidant content of three native Ecuadorian fruits: mora de monte (Rubus glabratus Kunth), mortiño (Vaccinium floribundum Kunth), and tuna de monte (Opuntia soederstromiana). Determination of resveratrol, ascorbic acid, citric acid, and sugars was determined by HPLC analysis, and UPLC analysis was used to determine tentative metabolites with nutraceutical properties. Antioxidant capacity was assessed using cyclic voltammetry and the DPPH method; differential pulse voltammetry was used to evaluate antioxidant power. Analysis of results through UPLC-QTOF mass spectrometry indicated that R. glabratus Kunth and V. floribundum Kunth are important sources of various compounds with potential health-promoting functions in the body. The DPPH results showed the following antioxidant capacities for the three fruits: R. glabratus Kunth > O. soederstromiana > V. floribundum Kunth; this trend was consistent with the antioxidant capacity results determined using the electrochemical methods.

FLNA expression modulates pathological markers of pituitary neuroendocrine tumours.

Due to the current limited knowledge about the role of filamin A (FLNA) in pituitary tumour progression, we aimed to analyse FLNA expression levels and its impact on aggressive markers of pituitary neuroendocrine tumours (PitNETs), using an integrative approach of in vivo and in vitro models and human samples. An increase in the expression levels of FLNA was observed in the advanced tumoural stages of the hyperplastic adenomatous pituitary model, concomitant with a decrease in cell proliferation and with a modification in the subcellular localisation of this protein. Similarly, overexpression of FLNA in the somatolactotropic GH3 cell line induced a decrease in the cell proliferation, promoted a migratory phenotype, increased invasion activity, and decreased the prolactin secretion. Cyclin D1 (CCND1) and cyclin-dependent kinase 4 (CDK4) expression increased in both models in correlation with the increase observed in FLNA levels. When human tissues were analysed a significant increase of FLNA was observed in PitNETs compared to normal pituitary gland, with heterogeneous intracellular localisation. Higher levels of FLNA expression were observed in tumours with invasive characteristics. These results underline the crucial roles of FLNA as a modulator of pathological markers and as a potential prognostic marker in pituitary tumours.

Endophytic bacterial communities in ungerminated and germinated seeds of commercial vegetables.

Chile is a prominent seed exporter globally, but the seed microbiome of vegetables (46% of seeds) and its role in the early stages of plant growth have remained largely unexplored. Here, we employed DNA metabarcoding analysis to investigate the composition and putative functions of endophytic bacterial communities in ungerminated and germinated seeds of the commercial vegetables Apiaceae (parsley and carrot), Asteraceae (lettuce), Brassicaceae (cabbage and broccoli), and Solanaceae (tomato). Bacterial quantification showed 104 to 108 copies of the 16S rRNA gene per gram of ungerminated and germinated seeds. Alpha diversity analysis (e.g., Chao1, Shannon, and Simpson indices) did not indicate significant differences (Kruskal-Wallis test) between ungerminated and germinated seeds, except for Solanaceae. However, beta diversity (PCoA) analysis showed distinctions (Adonis test) between ungerminated and germinated seeds, except Apiaceae. Pseudomonadota and Bacillota were identified as the dominant and specialist taxa in both ungerminated and germinated seed samples. Chemoheterotrophy and fermentation were predicted as the main microbial functional groups in the endophytic bacterial community. Notably, a considerable number of the 143 isolated endophytic strains displayed plant growth-promoting traits (10 to 64%) and biocontrol activity (74% to 82%) against plant pathogens (Xanthomonas and Pseudomonas). This study revealed the high variability in the abundance, diversity, composition, and functionality of endophytic bacteria between ungerminated and germinated seeds in globally commercialized vegetables. Furthermore, potential beneficial endophytic bacteria contained in their seed microbiomes that may contribute to the microbiome of the early stages, development, growth and progeny of vegetables were found.

Environmentally relevant DEHP exposure during gestational and lactational period inhibits filamin a testicular expression.

Toxicological studies have revealed that DEHP exposure during pregnancy may induce developmental disorders, especially in male offspring, leading to morphological and functional alterations in the reproductive system by mechanisms that should be investigated. Thus, the aim of this work was to analyze the testicular toxicity induced by an environmentally relevant DEHP dose during development and its impact on FLNA, a protein that participates in the blood-testis barrier assembly. We used male Wistar rats exposed to DEHP during pregnancy and lactation. The results showed that DEHP exposure during development and lactation increased body weight, decreased gonadal weight and shortened anogenital distance. This phthalate induced morphological changes in the testis, suggestive of hypospermatogenesis. DEHP exposure decreased the number of FLNA positive cells and the expression of FLNA and claudin-1 in prepubertal testes. Furthermore, DEHP inhibited FLNA and claudin-1 protein expression in adult male rats. These results indicated that exposure to DEHP during gestation and lactation perturbed testis development and suggested that FLNA is a target protein of DEHP, possibly contributing to the phthalate-induced damage on BTB.

Regulation of FGF2-induced proliferation by inhibitory GPCR in normal pituitary cells.

Introduction: Intracellular communication is essential for the maintenance of the anterior pituitary gland plasticity. The aim of this study was to evaluate whether GPCR-Gαi modulates basic fibroblast growth factor (FGF2)-induced proliferative activity in normal pituitary cell populations. Methods: Anterior pituitary primary cell cultures from Wistar female rats were treated with FGF2 (10ng/mL) or somatostatin analog (SSTa, 100nM) alone or co-incubated with or without the inhibitors of GPCR-Gαi, pertussis toxin (PTX, 500nM), MEK inhibitor (U0126, 100µM) or PI3K inhibitor (LY 294002, 10 µM). Results: FGF2 increased and SSTa decreased the lactotroph and somatotroph BrdU uptak2e (p<0.05) whereas the FGF2-induced S-phase entry was prevented by SSTa co-incubation in both cell types, with these effects being reverted by PTX, U0126 or LY294002 pre-incubation. The inhibition of lactotroph and somatotroph mitosis was associated with a downregulation of c-Jun expression, a decrease of phosphorylated (p) ERK and pAKT. Furthermore, SSTa was observed to inhibit the S-phase entry induced by FGF2, resulting in a further increase in the number of cells in the G1 phase and a concomitant reduction in the number of cells in the S phases (p< 0.05), effects related to a decrease of cyclin D1 expression and an increase in the expression of the cell cycle inhibitors p27 and p21. Discussion: In summary, the GPCR-Gαi activated by SSTa blocked the pro-proliferative effect of FGF2 in normal pituitary cells via a MEK-dependent mechanism, which acts as a mediator of both anti and pro-mitogenic signals, that may regulate the principal effectors of the G1 to S-phase transition.

Characterization of a Clostridioides difficile ST-293 isolate from a recurrent infection in Argentina

Resumen Clostridioides difficile es un patógeno esporulado oportunista responsable de diarrea asociada a antibióticos en humanos. C. difficile produce 2 toxinas principales: TcdAy TcdB, además de la toxina binaria (CDT), también asociada a la virulencia. Este estudio buscó caracterizar el aislamiento ALCD3, involucrado en un episodio de recurrencia de una infección nosocomial. La caracterización molecular mostró que dicho aislamiento pertenece al toxinotipo 0/v y el análisis por MLST demostró un perfil alélico adk:91, atpA:1, dxr:2, glyA: 1, recA:27, sodA: 1 y tpi:1, lo cual corresponde al ST293 (MLST clado 1). Durante el crecimiento, el aislamiento ALCD3 mostró un incremento temprano de la tasa de esporulación y valores máximos de formas termorresistentes luego de 2 días de incubación. Tanto la cinética de esporulación como la producción de formas termorresistentes fueron más rápidas en el aislamiento ALCD3 que en la cepa de referencia VPI 10463. La germinación en presencia del germinante natural taurocolato fue más rápida en el aislamiento ALCD3 que en la cepa VPI 10463, lo que indica que aquel comienza la hidrólisis del córtex antes. También, el co-germinante glicina indujo una rápida liberación de ácido dipicolínico en ALCD3. Estos hallazgos indican que el aislamiento ALCD3 es particularmente eficiente en la esporulación y en la germinación. El presente trabajo representa el primer informe de la circulación de C. difficile ST293 en Argentina. La habilidad del aislamiento ALCD3 para producir toxinas y su alta capacidad de esporulación/germinación son características claves compatibles con un alto potencial de diseminación e inducción de infecciones recurrentes.

Abstract Clostridioides difficile is an opportunistic spore-forming pathogen responsible for antibiotic-associated diarrhea in humans. C. difficile produces two main toxins: TcdA and TcdB as well as a third toxin named binary toxin (CDT) that is also involved in virulence. The present study aimed at characterizing the C. difficile isolate ALCD3 involved in a relapse episode of nosocomial infection. Molecular characterization showed that isolate ALCD3 belongs to tox-inotype 0/v and the MLST analysis demonstrated allelic profile adk:91, atpA:1, dxr:2, glyA: 1, recA:27, sodA: 1 and tpi:1 which corresponds to ST293 (MLST clade: 1). During growth, isolate ALCD3 showed an early increase in the sporulation ratio as well as maximal values of heat resis-tant forms after 2 days of incubation. Both sporulation kinetics and production of heat resistant forms were faster for isolate ALCD3 than for the reference strain VPI 10463. Germination in the presence of the natural germinant taurocholate was faster for isolate ALCD3 than for strain VPI 10463, which indicates that isolate ALCD3 starts cortex hydrolysis earlier than strain VPI 10463. Furthermore, the co-germinant glycine, induces rapid release of dipicolinic acid (DPA) in isolate ALCD3. These findings indicate that isolate ALCD3 is particularly efficient in both sporulation and germination. The present work represents the first report of the circulation of C. difficile ST293 in Argentina. The ability of isolate ALCD3 to produce toxins and its high sporulation/germination capacity are key features compatible with a microorganism with high dissemination potential and the possibility of inducing recurrent infections.

Nationwide genetic analysis of more than 600 families with inherited eye diseases in Argentina.

This study corresponds to the first large-scale genetic analysis of inherited eye diseases (IED) in Argentina and describes the comprehensive genetic profile of a large cohort of patients. Medical records of 22 ophthalmology and genetics services throughout 13 Argentinian provinces were analyzed retrospectively. Patients with a clinical diagnosis of an ophthalmic genetic disease and a history of genetic testing were included. Medical, ophthalmological and family history was collected. A total of 773 patients from 637 families were included, with 98% having inherited retinal disease. The most common phenotype was retinitis pigmentosa (RP, 62%). Causative variants were detected in 379 (59%) patients. USH2A, RPGR, and ABCA4 were the most common disease-associated genes. USH2A was the most frequent gene associated with RP, RDH12 early-onset severe retinal dystrophy, ABCA4 Stargardt disease, PROM1 cone-rod dystrophy, and BEST1 macular dystrophy. The most frequent variants were RPGR c.1345 C > T, p.(Arg449*) and USH2A c.15089 C > A, p.(Ser5030*). The study revealed 156/448 (35%) previously unreported pathogenic/likely pathogenic variants and 8 possible founder mutations. We present the genetic landscape of IED in Argentina and the largest cohort in South America. This data will serve as a reference for future genetic studies, aid diagnosis, inform counseling, and assist in addressing the largely unmet need for clinical trials to be conducted in the region.

Genetic variability of the 16S rRNA gene of Nocardia brasiliensis, the most common causative agent of actinomycetoma in Latin America and the Caribbean.

Mycetoma is a neglected tropical disease (NTD) declared by the World Health Organization (WHO) in 2016. It is characterized by the progressive growth of nodules and granulomatous lesions on the legs, arms, and trunk. It is potentially disfiguring and causes disability or amputations in working-age people from marginalized areas. The causative agents can be fungi (eumycetoma) or actinobacteria (actinomycetoma), the latter being the most common in America and Asia. Nocardia brasiliensis is the most important causal agent of actinomycetoma in the Americas. Taxonomic problems have been reported when identifying this species, so this study aimed to detect the 16S rRNA gene variations in N. brasiliensis strains using an in silico enzymatic restriction technique. The study included strains from clinical cases of actinomycetoma in Mexico, isolated from humans and previously identified as N. brasiliensis by traditional methods. The strains were characterized microscopically and macroscopically, then subjected to DNA extraction and amplification of the 16S rRNA gene by PCR. The amplification products were sequenced, and consensus sequences were constructed and used for genetic identification and in silico restriction enzyme analysis with the New England BioLabs® NEBcutter program. All study strains were molecularly identified as N. brasiliensis; however, in silico restriction analysis detected a diversity in the restriction patterns that were finally grouped and subclassified into 7 ribotypes. This finding confirms the existence of subgroups within N. brasiliensis. The results support the need to consider N. brasiliensis as a complex species.

Clostridioides difficile: Characterization of the circulating toxinotypes in an Argentinean public hospital / Clostridioides difficile: caracterización de los toxinotipos circulantes en un hospital público de Argentina

Abstract Clostridioides difficile is a spore-forming anaerobe microorganism associated to nosocomial diarrhea. Its virulence is mainly associated with TcdA and TcdB toxins, encoded by their respective tcdA and tcdB genes. These genes are part of the pathogenicity locus (PaLoc). Our aim was to characterize relevant C. difficile toxinotypes circulating in the hospital setting. The tcdA and tcdB genes were amplified and digested with different restriction enzymes: EcoRI for tcdA; HincII and AccI for tcdB. In addition, the presence of the cdtB (binary toxin) gene, TcdA and TcdB toxins by dot blot and the cytotoxic effect of culture supernatants on Vero cells, were evaluated. Altogether, these studies revealed three different circulating toxinotypes according to Rupnik's classification: 0, I and VIII, being the latter the most prevalent one. Even though more studies are certainly necessary (e.g. sequencing analysis), it is worth noting that the occurrence of toxinotype I could be related to the introduction of bacteria from different geographical origins. The multivariate analysis conducted on the laboratory values of individuals infected with the most prevalent toxinotype (VIII) showed that the isolates associated with fatal outcomes (GCD13, GCD14 and GCD22) are located in regions of the biplots related to altered laboratory values at admission. In other patients, although laboratory values at admission were not correlated, levels of urea, creatinine and white blood cells were positively correlated after the infection was diagnosed. Our study reveals the circulation of different toxinotypes of C. difficile strains in this public hospital. The variety of toxinotypes can arise from pre-existing microorganisms as well as through the introduction of bacteria from other geographical regions. The existence of microorganisms with different pathogenic potential is relevant for the control, follow-up, and treatment of the infections.

Resumen Clostridioides difficile es un anaerobio esporulado que se asocia con episodios de diarreas hospitalarias. Su virulencia se encuentra vinculada, principalmente, a las toxinas TcdA y TcdB, codificadas por sus respectivos genes, tcdA y tcdB, que son parte de un locus de patogenicidad (PaLoc). Nuestro objetivo fue caracterizar los toxinotipos de C. difficile circulantes en un hospital público. Los genes tcdA y tcdB fueron amplificados y digeridos con diferentes enzimas de restricción: EcoRI para tcdA; HincII y AccI para tcdB. Además, se evaluó la presencia de cdtB (gen de la toxina binaria B) y de las toxinas A y B (por dot blot), así como el efecto citotóxico de sobrenadantes de cultivo sobre células Vero. En conjunto, estos estudios revelaron tres toxinotipos circulantes según la clasificación de Rupnik: 0, I y VIII; el más prevalente fue el último. Aunque son necesarios más estudios (ej., secuenciación), es interesante notar que la presencia del toxinotipo I podría estar relacionada con la introducción de bacterias de diferente origen geográfico. En los pacientes infectados con el toxinotipo VIII, el análisis multivariante de los resultados de laboratorio mostró que los aislamientos asociados a decesos (GCD13, GCD14 y GCD22) estaban situados en regiones de los biplots relacionados con valores de laboratorio alterados al momento de la internación. En los otros pacientes, aunque no se observó correlación entre los valores de laboratorio al momento de la internación y la evolución clínica, los niveles de urea, creatinina y recuento de glóbulos blancos estuvieron correlacionados positivamente entre sí una vez diagnosticada la infección. Nuestro estudio revela la circulación de diferentes toxinotipos de C. difficile en un mismo hospital público. La variedad de toxinotipos puede originarse a partir de microorganismos preexistentes en la región, así como también por la introducción de bacterias provenientes de otras regiones geográficas. La existencia de microorganismos con diferente potencial patogénico es relevante para el control, el seguimiento y el tratamiento de las infecciones.

Antioxidant Capacity through Electrochemical Methods and Chemical Composition of Oenocarpus bataua and Gustavia macarenensis from the Ecuadorian Amazon.

This study evaluated the antioxidant properties and chemical composition of the seeds, pulp and peels of Ungurahua (Oenocarpus bataua) and Pasu (Gustavia macarenensis)-fruits, native to the Ecuadorian Amazon. The antioxidant capacity was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and cyclic voltammetry (antioxidant index 50 (AI50)) assays; differential pulse voltammetry was used to evaluate antioxidant power using the electrochemical index. The total phenolic content, as well as the yellow flavonoid and anthocyanin content, were quantified via spectrophotometry. In addition, the trans-resveratrol and ascorbic acid content were evaluated through high performance liquid chromatography (HPLC). Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) was used to identify secondary metabolites with possible therapeutic properties. Results showed that the Pasu peel and seed extracts had the highest antioxidant capacity, followed by the Ungurahua peel; these results were consistent for both spectroscopic and electrochemical assays. HPLC and UPLC-MS analysis suggest that Oenocarpus bataua and Gustavia macarenensis are important sources of beneficial bioactive compounds.

Ultrastructural and Molecular Evidence of Macroautophagy in Functioning PitNETs and Experimental Pituitary Tumors.

INTRODUCTION: Macroautophagy is a lysosome-mediated degradation process that controls the quality of cytoplasmic components and organelles, with its regulation depending on autophagy-related proteins (Atg) and with Beclin1/Atg6 and microtubule-associated protein light chain 3 (LC3/Atg8) being key players in the mammalian autophagy. As reports on this mechanism in the field of pituitary neuropathology and neuroendocrinology are scarce, our study analyzed the ultrastructural signs of macroautophagy and the expression of Beclin1 and LC3 proteins in human functioning PitNETs and in experimental pituitary tumors. METHODS: A group of humans functioning PitNETs and an experimental lactotroph model in rats of the F344 strain stimulated with estradiol benzoate (BE) were used. Ultrastructural and molecular evidence of the macroautophagic process was evaluated using different techniques. RESULTS: In functioning PitNETs cohort, 60% exhibited evidence of macroautophagy, with a significant difference found for Beclin1 and LC3 between macro- and micro-PitNETs (p < 0.05). In the experimental model, the expression of both Beclin1 and LC3 proteins was immunopositive in normal and tumoral glands when analyzed by immunofluorescence, Western blot, and immunohistochemistry. In the experimental model, protein expression was associated with increased glandular size and weight. CONCLUSIONS: Our study revealed evidence of macroautophagy at the pituitary level and the important role of Beclin1 and LC3 in the progression of functioning PitNETs, implying that this mechanism participate in regulating pituitary cell growth.

Characterization of a Clostridioides difficile ST-293 isolate from a recurrent infection in Argentina.

Clostridioides difficile is an opportunistic spore-forming pathogen responsible for antibiotic-associated diarrhea in humans. C. difficile produces two main toxins: TcdA and TcdB as well as a third toxin named binary toxin (CDT) that is also involved in virulence. The present study aimed at characterizing the C. difficile isolate ALCD3 involved in a relapse episode of nosocomial infection. Molecular characterization showed that isolate ALCD3 belongs to toxinotype 0/v and the MLST analysis demonstrated allelic profile adk:91, atpA:1, dxr:2, glyA: 1, recA:27, sodA: 1 and tpi:1 which corresponds to ST293 (MLST clade: 1). During growth, isolate ALCD3 showed an early increase in the sporulation ratio as well as maximal values of heat resistant forms after 2 days of incubation. Both sporulation kinetics and production of heat resistant forms were faster for isolate ALCD3 than for the reference strain VPI 10463. Germination in the presence of the natural germinant taurocholate was faster for isolate ALCD3 than for strain VPI 10463, which indicates that isolate ALCD3 starts cortex hydrolysis earlier than strain VPI 10463. Furthermore, the co-germinant glycine, induces rapid release of dipicolinic acid (DPA) in isolate ALCD3. These findings indicate that isolate ALCD3 is particularly efficient in both sporulation and germination. The present work represents the first report of the circulation of C. difficile ST293 in Argentina. The ability of isolate ALCD3 to produce toxins and its high sporulation/germination capacity are key features compatible with a microorganism with high dissemination potential and the possibility of inducing recurrent infections.

Clostridioides difficile: Characterization of the circulating toxinotypes in an Argentinean public hospital.

Clostridioides difficile is a spore-forming anaerobe microorganism associated to nosocomial diarrhea. Its virulence is mainly associated with TcdA and TcdB toxins, encoded by their respective tcdA and tcdB genes. These genes are part of the pathogenicity locus (PaLoc). Our aim was to characterize relevant C. difficile toxinotypes circulating in the hospital setting. The tcdA and tcdB genes were amplified and digested with different restriction enzymes: EcoRI for tcdA; HincII and AccI for tcdB. In addition, the presence of the cdtB (binary toxin) gene, TcdA and TcdB toxins by dot blot and the cytotoxic effect of culture supernatants on Vero cells, were evaluated. Altogether, these studies revealed three different circulating toxinotypes according to Rupnik's classification: 0, I and VIII, being the latter the most prevalent one. Even though more studies are certainly necessary (e.g. sequencing analysis), it is worth noting that the occurrence of toxinotype I could be related to the introduction of bacteria from different geographical origins. The multivariate analysis conducted on the laboratory values of individuals infected with the most prevalent toxinotype (VIII) showed that the isolates associated with fatal outcomes (GCD13, GCD14 and GCD22) are located in regions of the biplots related to altered laboratory values at admission. In other patients, although laboratory values at admission were not correlated, levels of urea, creatinine and white blood cells were positively correlated after the infection was diagnosed. Our study reveals the circulation of different toxinotypes of C. difficile strains in this public hospital. The variety of toxinotypes can arise from pre-existing microorganisms as well as through the introduction of bacteria from other geographical regions. The existence of microorganisms with different pathogenic potential is relevant for the control, follow-up, and treatment of the infections.

A novel kinetic model for a cocoa waste fermentation to ethanol reaction and its experimental validation.

A non-segregated kinetic model is proposed to describe a fermentation process of agro-industrial residues derived via cocoa (mucilage juice) by Pichia kudriavzevii. The novel proposed hybrid model is based on a multiple coupling reaction mechanisms (structured) to describe the kinetics of substrate consumption, biomass, carbon dioxide, and ethanol, coupled to an unstructured model for the activity enzyme. The parameters of the kinetic model are estimated by non-linear least-squares curve fitting using the Marquardt-Levenberg algorithm. In addition, numerical simulations were compared with the experimental data via residual graphs. The effectiveness of the model was statistically evaluated using dimensionless efficiency coefficients under different initial conditions. A global sensitivity analysis was applied (Fisher's information matrix). The experimental results of the batch reactor showed a maximum ethanol concentration of 29 g/L, with a yield of 0.48 g-ethanol/g-glucose and a productivity of 0.30 g/L h. The method determined that the cell formation coefficient and the specific substrate consumption rate (θ1 and θ2) directly influence most of the states of our system. The proposed scheme is particularly suitable to assist in the rational design of cell factory properties or fermentation processes because it can represent the complex biochemistry in more detail and under different initial experimental conditions; the above reveals that the generated model is robust and can be considered for control and optimization purposes.

Rol de la Comisión Honoraria para la Lucha Antituberculosa y Enfermedades Prevalentes en la Estrategia Nacional de Vacunación contra la COVID-19 en Uruguay: período noviembre 2020 - julio 2021 / Role of the Honorary Commission for the Fight against Tuberculosis and Prevalent Diseases (CHLA-EP) in the National Vaccination Strategy against COVID-19 in Uruguay: november 2020 - July 2021 / Papel da Comissão Honorária de Luta contra a Tuberculose e Doenças Prevalentes (CHLA-EP) na Estratégia Nacional de Vacinação contra a COVID-19 no Uruguai: período de novembro de 2020 a julho de 2021

La pandemia por la infección por el nuevo coronavirus SARS-CoV-2 determinó que en Uruguay a inicio de marzo de 2020 el gobierno diseñara el Plan Nacional de Contingencia para la enfermedad COVID-19 causada por este virus. Este plan incluyó medidas preventivas no farmacológicas como: higiene de manos, distanciamiento físico, uso de mascarilla de diferentes tipos de acuerdo al riesgo de quien debía utilizarlos, en algunos períodos restricciones varias sobre circulación de personas, actividades educativas o espectáculos públicos, y tránsitos por tierra, aire o mar. Se apeló fuertemente a la libertad responsable. A partir de diciembre 2020 se decidió incorporar a la prevención primaria una Estrategia de Vacunación. El 1/3/2021 se inició la Campaña Vacunación COVID-19 2021, que fue no obligatoria, poblacional, inicialmente para personas mayores de 18 años y a partir de junio incluyó adolescentes entre 12 y 17 años y mujeres embarazadas. El Grupo Asesor Científico Honorario (GACH) del Gobierno Nacional junto a la Comisión Nacional Asesora de Vacunas (CNAV) del Ministerio de Salud Pública (MSP) que cuenta con largos años de aconsejar a las autoridades sanitarias en decisiones científicas sobre la introducción de vacunas, y una Comisión ad Hoc creada para vacunas COVID-19 reunieron a expertos del país para asesorar sobre la estrategia de vacunación. La Comisión Honoraria para la Lucha Antituberculosa y Enfermedades Prevalentes (CHLA-EP) como brazo operativo del Departamento de Inmunizaciones del MSP estuvo desde el mes de diciembre involucrada con todos los actores para implementar la Campaña de Vacunación poblacional COVID-19 sin descuidar las tareas que le competen en la vacunación para prevenir otras enfermedades infecciosas. Metodología: se realizó un estudio descriptivo, retrospectivo de las estrategias implementadas por el Dpto. Operativo de Inmunizaciones (brazo operativo del Departamento de Inmunizaciones del MSP), Laboratorio A. Calmette, Comisión Honoraria, Coordinación General y los departamentos administrativos: entre los meses de noviembre de 2020 a julio de 2021. Fuente de datos: informes difundidos o aportados especialmente para este trabajo por el Dpto. Operativo de Inmunizaciones, Laboratorio A. Calmette, Dpto. de Inmunizaciones del Ministerio de Salud Pública, Recursos Humanos, Recursos Materiales, Servicio de Apoyo, Comunicación y Diseño, Centros Periféricos, Dirección Ejecutiva y Secretaría General. Resultados: se muestran los resultados de 4 períodos: 1) planificación (noviembre-diciembre 2020); 2) capacitación, contratación de recursos humanos y asesoramiento para la adquisición de recursos materiales (enero-febrero 2021); 3) apertura de centros (hubs), puestos de vacunación COVID-19 y puesta en marcha de la Campaña COVID-19 (marzo 2021); 4) superposición campaña antigripal 2021 (abril-junio 2021) y mantenimiento coberturas de las vacunas del (PNI) 1. Planificación: se tomaron en cuenta las características de las vacunas que se recibirían vacunas a virus muertos, vacunas vectoriales y en particular la RNAm con requerimiento de almacenamiento a -70ºC, traslado y distribución a -25ºC, y reconstitución y uso a +28ºC en los diferentes escenarios. Se realizó análisis de riesgo y análisis de costo. Se intercambió con diferentes empresas que brindan dicho servicio, participando de actividades de capacitación con alguna de ellas, fundamentales a la hora de diseñar una cadena de frío sólida y sin fallas (por ejemplo actividad de entrenamiento con Vac-Q-tec). Se diseñó el traslado de las vacunas hacia los hubs, vacunatorios terciarios o centros periféricos de vacunación. Teniendo en cuenta el sitio de llegada de las vacunas, se plantearon los diferentes escenarios posibles, no solo a nivel central (Montevideo-Aeropuerto Internacional de Carrasco), sino también a nivel nacional utilizando las diferentes terminales aéreas. Así también se planificaron estrategias de distribución de las vacunas por vía terrestre involucrando diferentes actores (públicos o privados), basados en la experiencia de la institución debido al cotidiano trabajo que realizan, tanto en campaña de vacunación como en el mantenimiento del PNI. Se realizó la previsión de compras de insumos para el almacenamiento y distribución de la vacuna, como para la vacunación en sí. Se trabajó en conjunto con Departamento de Compras de Ministerio de Economía y Dirección de Secretaria del MSP. Se participó en la conformación de equipos de vacunación con la consigna de vacunación rápida, universal, gratuita y segura. Se estableciendo roles por equipo de vacunación constituidos por 6 personas. 2. Capacitación, contratación de recursos humanos y asesoramiento para la adquisición de recursos materiales. Se confeccionaron cursos y materiales en calidad y cantidad para que el personal adquiriera conocimientos actualizado sobre SARS-CoV-2 y COVID-19, mecanismos de transmisión, utilización y planificación de uso de equipos de protección personal (EPP) y seguimiento detallado de las vacunas en desarrollo. Análisis minucioso de los posibles candidatos a llegar a Uruguay, focalizándose en tres de ellos (Pfizer BioNtech, CoronaVac, Oxford AZ). Se realizó capacitación por roles (vacunadores, preparadores, administradores, referente de puesto, backup). Especial énfasis tuvo la capacitación en el cuidado de la persona a vacunar, registro del acto vacunal, cadena de frío en todos sus puntos, descarte de materiales utilizados y llenado de planillas de fiscalización. Todos los contenidos y materiales quedaron disponibles en la página web de la institución. Recursos humanos: se destinaron funcionarios para la confección, firma y papeleo de altas de 1.463 nuevos contratos de los equipos de todo el país, RRHH dedicados no sólo a la vacunación en sí, sino a participar activamente en cada uno de los puestos de vacunación. Esto significó que el trabajo se multiplicó por seis para las tareas requeridas durante este período. El Dpto. Recursos Materiales pasó de empaquetar y distribuir de 7 a 15 paquetes a 80 paquetes mensuales para cada centro periférico/vacunatorio. El envío habitual de materiales se cuadruplicó y en los comienzos de la campaña se llegó a quintuplicar y sextuplicar. Algunas de las cifras a destacar son: 3.300.000 jeringas con aguja, 8.800 Litros de alcohol eucaliptado, 852.000 guantes de látex y nitrilo y 141.000 sobreáticos. 3. Apertura de centros (hubs), puestos de vacunación COVID-19 y puesta en marcha de la Campaña COVID-19. Esto generó en forma progresiva la habilitación de centros (hubs) de máxima respuesta (vacunación masiva): ocho en Montevideo, uno en Canelones y uno en Maldonado. Dado que se decidió vacunar de 3 a 5 personas cada 15 minutos, en el hub del Hospital de Clínicas se administaron 211.153 dosis en sus 32 puestos de vacunación y en el hub Antel Arena se administraron 615.637 dosis en sus 15 puestos de vacunación. Al 31 de julio habían 144 puestos de vacunación COVID 19 habilitados en todo el territorio Nacional. La distribución se realizó a nivel nacional: 3.229. 320 dosis de vacuna Sinovac, 97.790 dosis de vacunas AstraZeneca y 1.585452 de vacuna Pfizer. Se participó en la conformación de 60 equipos con un referente (identificación, registro, control), preparador, 2 vacunadores y chofer para la vacunación en los Establecimientos de Larga Estadía de Personas Mayores (ELEPEM) y personas con situaciones especiales. Se participó activamente en la vacunación en ELEPEM, refugios y escuelas especiales, visitándose un total de 1679 Instituciones y se vacunaron 35.006 personas. 4. Superposición campaña antigripal 2021 (abril-junio 2021) y mantenimiento coberturas de las vacunas del (PNI). A partir del 24/04/2021 comenzó la Campaña Antigripal 2021, superponiéndose a la campaña descripta de COVID-19, lo que implicó reforzar las estrategias y ampliar las acciones para poder cumplir con ambas campaña paralelamente, siempre con el objetivo de no desatender el Programa Nacional de Inmunizaciones. Dentro de ésta se llevó a cabo la vacunación en los ELEPEM, como anualmente realiza la comisión y vacunación en territorio de vacunas COVID-19, antigripal y del PNI. Según los datos recibidos al 17 de Julio en el informe de la Unidad de Inmunizaciones del MSP (semana 12), fueron 605.802 dosis registradas, de las cuales al menos 87.802 dosis fueron administradas por la institución. Esta diferencia en relación a las campañas anteriores podría ser atribuida al intenso trabajo realizado en la estrategia de vacunación COVID-19. En el marco de la actividad interinstitucional barrio a barrio las personas vacunadas con vacuna antigripal en el móvil de la CHLA-EP en Montevideo fueron 7.225. La CHLA-EP ha contribuido desde la logística y la vacunación con esquema de dos dosis para las tres plataformas y una dosis de refuerzo para los que recibieron vacunas a virus muertos. La CHLA-EP ha contribuido para alcanzar las cifras de la Campaña de Vacunación COVID-19 entre el 1/3 y el 29/8/21: 5.487.181 dosis aplicadas, 2.65.269 personas que recibieron una dosis de vacuna y de ellas 2.496.290 tienen dos dosis y han recibido dosis de refuerzo 333.662. Conclusiones: la planificación, capacitación, provisión de recursos humanos y materiales lograron llevar a cabo dos campañas de vacunación coexistentes en hubs, vacunatorios, ELEPEM y en territorio que se han considerado exitosas e históricas.

The new Coronavirus SARS CoV-2 pandemic was declared at the beginning of March 2020 when the Uruguayan government appointed the National Contingency Plan for the COVID-19 disease caused by this virus. This plan included non-pharmacological preventive measures such as: hand hygiene, physical distancing, use of different types of masks according to the risk of who should wear them, in some periods, restrictions to mobility, educational activities or public shows, and movement by land, air or sea. Responsible freedom was strongly called for. In December 2020, the Vaccination Strategy was included as a primary prevention mechanism. On 3/1/2021, the COVID-19 2021 Vaccination Campaign began, it was non-mandatory, population-based, initially for people over 19 years of age, and for adolescents between 12 and 17 years of age and pregnant women since June. The National Honorary Scientific Advisory Group (GACH) together with the National Vaccine Advisory Committee (CNAV) of the Ministry of Public Health (MSP) that has vast experience in advising health authorities in scientific decisions on the introduction of vaccines, and an ad-Hoc Commission created for COVID-19 vaccines, brought together experts to advise on the vaccination strategy. The Honorary Commission for the Fight Against Tuberculosis and Prevalent Diseases (CHLA-EP), as the operational branch of the MSP's Department of Immunizations, has participated with all stakeholders since December to implement the COVID-19 Population Vaccination Campaign without neglecting their tasks regarding the vaccination to prevent other infectious diseases. Methodology: a descriptive, retrospective study was carried out regarding the strategies implemented by the Immunization Operational Department (operational branch of the Ministry of Health's Immunization Department), A. Calmette Laboratory, Honorary Commission, General Coordination and Administrative Departments between November, 2020 and July 2021. Data source: reports were issued or provided specially for this work by the Immunization Operational Department, A. Calmette Laboratory, by the Ministry of Public Health's immunization Department, Human Resources, Material Resources, Support, Communication and Design Service, Peripheral Centers, Executive Board and General secretariat. Results: the results of the 4 periods are as follows: 1- Planning (Period November - December 2020), 2- Training, hiring of human resources and advice for the acquisition of qualitative and quantitative material resources (January - February 2021), 3- Opening of COVID-19 vaccination centers (Hubs), posts and launch of the COVID-19 Campaign (March 2021), 4- Overlap of the 2021 flu campaign (April-June) and maintenance of vaccine coverage (National Immunization Programme - PNI). 1- Planning: The characteristics of the different vaccines (dead virus vaccines, vector vaccines and in particular mRNA vaccines, with storage requirements of -70ºC, transfer and distribution requirements of -25ºC, and reconstitution and use of +2 + 8ºC) were taken into account in the different scenarios. Risk and costs analysis were performed. We interchanged data with different companies that provided this service, we participated in training activities with some of them, mainly in the design of a solid cold chain without failures (e.g. a Vac-Q-tec training activity). The transfer of vaccines to hubs, tertiary vaccination centers or peripheral vaccination centers was designed. The different destination site of the vaccines, the different possible scenarios were taken into account, both at local central level (Montevideo - Carrasco International Airport), but also at National level using the different air terminals. Similarly, strategies for the distribution of vaccines by land were planned involving different public or private actors based on the institutions' experience, both regarding the vaccination campaign and in the PNI maintenance. We made a supply acquisition plan for the vaccine's storage and distribution, for the vaccination itself, and for the purchase of supplies jointly with the Procurement Department of the Ministry of Economy and Executive Secretariat of the MSP. Six-people vaccination teams were created with clear roles and with the slogan of rapid, universal, free and safe vaccination for all. 2- Training: Courses and materials were organized for the staff to acquire updated knowledge about SARS CoV 2 and COVID-19, transmission mechanisms, use and planning of use of personal protective equipment (PPE) and of course, a detailed monitoring of the vaccines being developed. We carried out a thorough analysis of the vaccine companies to arrive in Uruguay, we focused on 3 of them (Pfizer BioNtech, CoronaVac, Oxford AZ). Training was carried out by roles (vaccinators, preppers, administrators, position referents, backups). Special emphasis was placed on the training of the care of the person to be vaccinated, registration of the vaccination act, end-to-end cold chain, disposal of used materials and filing out inspection forms. All the contents and materials were available on the Institution's webpage. Human resources: human resources were allocated for the creation, signature and registration of the paperwork for 1,463 new contracts for teams across the country. These resources were allocated not only to vaccination itself, but also to actively participate in each of the vaccination posts. This meant that the work was multiplied by 6 for the tasks required during this period. The Department of Material Resources, went from packaging and distributing from 7 to 15 packages a month to 80 packages a month for each Peripheral/Vaccination Center. Material shipment quadrupled at the beginning of the campaign and it eventually grew by five and sixfold. Some of the key figures to highlight are: 3, 300,000 needle syringes, 8,800 liters of Eucalyptus Alcohol, 852,000 latex and nitrile gloves and 141,000 isolation gowns. 3. COVID-19 Vaccination Hubs, Posts and Campaigns. We gradually opened maximum response mass vaccination Hubs in Montevideo (8), Canelones (1) and Maldonado (1). Since it was had been decided to vaccinate 3 to 5 people every 15 minutes, the primary results at the Hospital de Clínicas Hub were 211,153 doses administered in 32 vaccination posts, and at Antel Arena Hub 615,637 doses were administered in its 15 vaccination posts. As of July 31, there were 144 COVID 19 vaccination posts throughout the national territory. The distribution was made nationwide: 3,229,320 Sinovac vaccine doses, 97,790 Oxford-Astra Zeneca vaccine doses, and 1,585,452 Pfizer-BioNtech vaccine doses. We devised 60 teams including a referent (identification, registration, control), a trainer, 2 vaccinators and vaccination driver for Homes for the Elderly at ELEPEM and people needing special care. They actively participated in the vaccination at ELEPEM, Shelters and Schools for Children with Special Needs and visited a total of 1679 Institutions and overall, 35,006 people were vaccinated. 4- Overlap of the 2021 flu campaign (April-June) and the maintenance of the COVID vaccine coverage (National Immunization Programme - PNI). Since 04/24/2021, the Anti-Flu Campaign began, and it overlapped the COVID-19 campaign described above, which meant reinforcing strategies and expanding actions to be able to carry out both campaigns simultaneously, always with the objective of maintaining the National Immunization Program. Vaccination was carried out at the ELEPEM, as it is performed annually by the commission and vaccination of COVID-19, influenza and PNI was performed on the territory. According to the data received on July 17 in the Ministry of Public Health Immunization Unit Report (week 12), there were 605,802 registered doses, of which at least 87,802 had been administered by the Institution. This difference regarding previous campaigns could have been caused by the intense work carried out during the COVID19 Vaccination Campaign. Within the framework of the inter-institutional activity "Barrio a Barrio" (Vaccination in all Neighborhoods), 7,225 people were vaccinated with the influenza vaccine at the CHLA-EP mobile vaccination bus in Montevideo. The CHLA-EP has contributed with logistics and vaccination with a 2-dose schedule for the 3 platforms and a booster dose for those who received virus vaccines. The CHLA-EP has contributed to achieving the target of the COVID-19 Vaccination Campaign between 1/3 and 8/29/21: 5,487,181 doses were applied, 2,065,269 people received 1 dose of vaccine and 2,496,290 of them received 2 doses and 333,662 have received booster doses. Conclusion: the planning, training, provision of human and material resources has enabled us to successfully carry out two historical simultaneous vaccination campaigns in hubs, vaccination centers, ELEPEM and all along the territory.

A pandemia causada pela infecção pelo novo Coronavírus SARS CoV-2 determinou que no Uruguai no início de março de 2020 o governo elaborasse o Plano Nacional de Contingência para a doença COVID-19 causada por este vírus. Este plano incluía medidas preventivas não farmacológicas como: higiene das mãos, distanciamento físico, uso de diferentes tipos de máscaras dependendo do risco de quem as deve usar, em alguns períodos várias restrições à circulação de pessoas, atividades educativas ou espetáculos públicos e a trânsito por terra, ar ou mar). Foi feito um forte apelo à liberdade responsável. A partir de dezembro de 2020, decidiu-se incorporar uma Estratégia de Vacinação à prevenção primária. Em 01/03/2021 teve início a Campanha de Vacinação COVID-19 2021, não obrigatória, de base populacional, inicialmente para maiores de 18 anos e a partir de junho contempla adolescentes entre 12 e 17 anos e gestantes. O Grupo Honorário de Assessoramento Científico do Governo Nacional juntamente com a Comissão Nacional de Assessoramento de Vacinas (CNAV) do Ministério da Saúde Pública (MSP), que assessora autoridades de saúde em decisões científicas sobre a introdução de vacinas, e uma Comissão ad hoc criada para vacinas contra a COVID-19 reuniu especialistas do país para aconselhar sobre a estratégia de vacinação. A Comissão Honorária de Luta contra a Tuberculose e Doenças Prevalentes (CHLA-EP) como braço operacional do Departamento de Vacinação do MSP envolveu-se desde dezembro com todos os intervenientes na implementação da Campanha de Vacinação da População COVID-19, aliás de realizar as tarefas de vacinação para prevenir outras doenças infecciosas. Metodologia: foi realizado um estudo descritivo e retrospectivo das estratégias implementadas pelo Departamento Operacional de Imunizações (braço operacional do Departamento de Imunizações do MSP), Laboratório A. Calmette, Comissão Honorária, Coordenação Geral e departamentos administrativos: entre os meses de novembro de 2020 a julho de 2021. Fonte de dados: relatórios divulgados ou oferecidos especialmente para este trabalho pelo Departamento Operacional de Imunizações, Laboratório A. Calmette, Departamento de Imunizações do Ministério da Saúde Pública, Recursos Humanos, Recursos Materiais, Serviço de Apoio, Comunicação e Design, Centros Periféricos, Direção Executiva e Secretaria Geral. Resultados: são apresentados os resultados de 4 períodos: 1- Planejamento (novembro - dezembro de 2020), 2- Capacitação, contratação de recursos humanos e assessoria para aquisição de recursos materiais em qualidade e quantidade (janeiro - fevereiro de 2021), 3- Abertura de centros (Hubs), postos de vacinação COVID-19 e implementação da Campanha COVID-19 (março 2021), 4- Sobreposição da campanha da gripe 2021 (abril-junho 2021) e manutenção da cobertura vacinal (PNI) 1- Planejamento: As características das vacinas a receber eram vacinas de vírus mortos, vacinas vetoriais e em particular mRNA com requisitos de armazenamento de temperaturas de -70ºC, transferência e distribuição de -25ºC, e reconstituição e uso de +2 +8ºC nos diferentes cenários. Análise de risco e análise de custo foram realizadas. Realizaram-se intercâmbios com diferentes empresas que prestam este serviço, participando em ações de formação com algumas delas, essenciais na criação de uma cadeia de frio sólida e sem falhas (ex. ação de formação com Vac-Q-tec). Foi projetada a transferência de vacinas para os hubs, centros de vacinação terciários ou periféricos. Tendo em conta o local de chegada das vacinas, foram considerados os diferentes cenários possíveis, não só a nível central (Montevidéu - Aeroporto Internacional de Carrasco), mas também a nível nacional utilizando os diferentes terminais aéreas. Da mesma forma, foram planejadas estratégias de distribuição de vacinas por via terrestre envolvendo diferentes atores (públicos ou privados), com base na experiência da instituição, tanto na campanha de vacinação quanto na manutenção do PNI. A previsão de compra de insumos foi feita tanto para o armazenamento e distribuição da vacina, quanto para a própria vacinação, como também na tomada de decisão na compra de insumos, atuando em conjunto com o Departamento de Compras do Ministério da Economia e Gestão Secretário do MSP. Eles participaram da formação de equipes de 6 pessoas com funções especificas de vacinação com o slogan de vacinação rápida, universal, gratuita e segura. 2- Capacitação, contratação de recursos humanos e assessoria para aquisição de recursos materiais em qualidade e quantidade. Foram organizados cursos e materiais para que os funcionários adquirissem conhecimento atualizado sobre SARS CoV2 e COVID-19, mecanismos de transmissão, uso e planejamento de uso de equipamentos de proteção individual (EPI) e também para o acompanhamento minucioso das vacinas em desenvolvimento. Fizemos a análise minuciosa das possíveis vacinas candidatas para chegar ao Uruguai, e focamos em 3 delas (Pfizer BioNtech, CoronaVac, Oxford AZ). O treinamento foi realizado por funções (vacinadores, treinadores, administradores, posição de referência, backup). Foi dado especial destaque à formação no cuidado da pessoa a vacinar, registo do ato vacinal, rede de frio em todos os seus pontos, eliminação de materiais utilizados e preenchimento de fichas de controlo. Todos os conteúdos e materiais foram disponibilizados na página web da Instituição. Recursos humanos: funcionários foram alocados para a preparação, assinatura e registro de 1.463 novos contratos para complementar as equipes em todo o país, estes recursos humanos estiveram dedicados não apenas à vacinação em si, mas a participar ativamente de cada um dos postos de vacinação. Isso significou que o trabalho se multiplicou por 6 para as tarefas exigidas durante esse período. O Departamento de Recursos Materiais, passou de embalar e distribuir de 7 para 15 embalagens por mês, passando para 80 embalagens para cada Periférico/Centro de Vacinação. A remessa habitual de materiais se quadruplicou e no início da campanha aumentou cinco e seis vezes. Alguns dos números a destacar são: 3.300.000 seringas com agulha, 8.800 litros de álcool de eucalipto, 852.000 luvas de látex e nitrílica e 141.000 túnicas. 3. Abertura de centros e postos de vacinação COVID-19 e lançamento da Campanha COVID-19. Isso gerou progressivamente o estabelecimento de centros de resposta máxima (Hubs) (vacinação em massa) 8 em Montevidéu, um em Canelones e um em Maldonado. Já que decidiu-se vacinar de 3 a 5 pessoas a cada 15 minutos, no Hub. O Hospital de Clínicas administrou 211.153 doses em seus 32 postos de vacinação e na Arena Hub Antel foram administradas 615.637 doses em seus 15 postos de vacinação. Até 31 de julho, havia 144 postos de vacinação contra a COVID 19 habilitados em todo o território nacional. A distribuição foi realizada, 3.229 foram distribuídos em todo o país, 320 doses de vacina Sinovac, 97.790 doses de vacinas AstraZeneca e 1.585452 de vacina Pfizer. Participaram no treinamento de 60 equipes de referência (identificação, registo, controle), treinador, 2 vacinadores e motorista para vacinação em estabelecimentos de longa permanência para idosos (ELEPEM) e pessoas com situações especiais em albergues e escolas especiais, visitando um total de 1.679 Instituições e vacinando a 35.006 pessoas. 4- Sobreposição da campanha de gripe de 2021 (abril a junho de 2021) e manutenção da cobertura vacinal (PNI). A partir de 24/04/2021 teve início a Campanha da Gripe 2021, sobrepondo-se à campanha COVID-19 descrita, o que implicou o reforço de estratégias e ampliação das ações para poder cumprir ambas campanhas em paralelo, sempre com o objetivo de não descurar o Programa Nacional de Imunizações. Foi realizada a vacinação no ELEPEM, conforme anualmente realizada pela comissão e a vacinação no território das vacinas COVID-19, influenza e PNI. De acordo com os dados recebidos até 17 de julho no relatório da Unidade de Imunizações dos Ministérios da Saúde Pública (semana 12), foram registradas 605.802 doses, das quais pelo menos 87.802 doses foram administradas pela Instituição. Essa diferença em relação às campanhas anteriores pode ser atribuída ao intenso trabalho realizado na estratégia de Vacinação COVID19. No âmbito da atividade interinstitucional Bairro a Bairro, 7.225 pessoas foram vacinadas com a vacina contra gripe no celular do CHLAEP de Montevidéu. O CHLA-EP tem contribuído desde a logística e vacinação com um esquema de 2 doses para as 3 plataformas e uma dose de reforço para quem recebeu vacinas de vírus mortos. O CHLA-EP contribuiu para atingir os números da Campanha de Vacinação COVID-19 entre 1/3 e 29/8/21: 5.487.181 doses aplicadas, 2.065.269 pessoas que receberam 1 dose de vacina e delas 2.496.290 têm 2 doses e 333.662 receberam doses de reforço. Conclusões: O planejamento, capacitação, disponibilização de recursos humanos e materiais permitiram a realização de duas campanhas de vacinação coexistentes nos hubs, vacinatórios, ELEPEM e no território, o que pode ser considerado um processo bem sucedido e histórico.

Genetic variability of the 16S rRNA gene of Nocardia brasiliensis, the most common causative agent of actinomycetoma in Latin America and the Caribbean

ABSTRACT Mycetoma is a neglected tropical disease (NTD) declared by the World Health Organization (WHO) in 2016. It is characterized by the progressive growth of nodules and granulomatous lesions on the legs, arms, and trunk. It is potentially disfiguring and causes disability or amputations in working-age people from marginalized areas. The causative agents can be fungi (eumycetoma) or actinobacteria (actinomycetoma), the latter being the most common in America and Asia. Nocardia brasiliensis is the most important causal agent of actinomycetoma in the Americas. Taxonomic problems have been reported when identifying this species, so this study aimed to detect the 16S rRNA gene variations in N. brasiliensis strains using an in silico enzymatic restriction technique. The study included strains from clinical cases of actinomycetoma in Mexico, isolated from humans and previously identified as N. brasiliensis by traditional methods. The strains were characterized microscopically and macroscopically, then subjected to DNA extraction and amplification of the 16S rRNA gene by PCR. The amplification products were sequenced, and consensus sequences were constructed and used for genetic identification and in silico restriction enzyme analysis with the New England BioLabs® NEBcutter program. All study strains were molecularly identified as N. brasiliensis; however, in silico restriction analysis detected a diversity in the restriction patterns that were finally grouped and subclassified into 7 ribotypes. This finding confirms the existence of subgroups within N. brasiliensis. The results support the need to consider N. brasiliensis as a complex species.

Kummel's disease

Kummel's disease is a crush fracture of a vertebral body caused by a minor trauma, with the most accepted etiology being avascular necrosis. It is more frequent in individuals with risk factors such as osteoporosis or prolonged treatment with corticosteroids. Initially, it usually has normal radiological tests and an asymptomatic period, followed by a progressive onset of pain along with probable kyphosis and a sign of void or fluid abscess on radiological tests, which may create nerve/spinal involvement. The case is presented of a 76-year-old man, who was admitted to this center for the differential diagnosis of a single vertebral lesion. After imaging tests and biopsy, the definitive diagnosis of Kummel's disease was reached. The confirmatory diagnosis was reached by vertebral biopsy, but given its invasive nature, imaging techniques can play a significant role. As regards metabolic imaging tests, bone scintigraphy has shown to be one of the most sensitive tools to detect ischemia in earlier stages or to determine if it affects other locations. The whole body scan with diphosphonates shows an increase in activity in relation to bone remodeling activity in this condition. The 3-phase study makes it possible to differentiate whether it is an acute/subacute or chronic process, and can influence the therapeutic decision. Knowledge of this disease is important to make a differential diagnosis with tumour or infectious pathology, with emphasis on performing imaging tests in the event of persistent pain with a normal initial plaque.

La enfermedad de Kummel es una fractura-aplastamiento de un cuerpo vertebral precedida por un traumatismo menor, cuya etiología más aceptada es la necrosis avascular. Es más frecuente en individuos con factores de riesgo como osteoporosis o tratamiento prolongado con corticoides. Inicialmente, suele presentar pruebas radiológicas normales y un periodo asintomático, con aparición progresiva del dolor junto a probable cifosis y signo del vacío o absceso líquido en las pruebas radiológicas, pudiendo llegar a crear compromiso nervioso/medular. Presentamos el caso de un varón de 76 arios que ingresa en nuestro centro para el diagnóstico diferencial de una lesión única vertebral; tras la realización de las pruebas de imagen y biopsia se llega al diagnóstico definitivo de enfermedad de Kummel. El diagnóstico de confirmación de esta enfermedad se alcanza mediante la biopsia vertebral, pero dada su naturaleza invasiva, las técnicas de imagen toman un papel relevante. En relación con las pruebas de imagen metabólicas, la gammagrafía ósea ha demostrado ser una de las herramientas más sensibles para detectar isquemia en fases más tempranas o para conocer si afecta a otras localizaciones. El rastreo corporal de cuerpo completo con difosfonatos muestra un aumento de actividad en relación con la actividad ósea remodelativa en este cuadro. El estudio de tres fases permite diferenciar si se trata de un proceso agudo/subagudo o crónico, lo que influye en la decisión terapéutica. Es importante el conocimiento de esta enfermedad para realizar diagnóstico diferencial con patología tumoral o infecciosa e insistir en la realización de pruebas de imagen ante la persistencia del dolor con una placa inicial normal.