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Sterol 14-demethylase (CYP51) inhibitors, encompassing new chemical entities and repurposed drugs, have emerged as promising candidates for Chagas disease treatment, based on preclinical studies reporting anti-Trypanosoma cruzi activity. Triazoles like ravuconazole (RAV) and posaconazole (POS) progressed to clinical trials. Unexpectedly, their efficacy was transient in chronic Chagas disease patients, and their activity was not superior to benznidazole (BZ) treatment. This paper aims to summarize evidence on the global activity of CYP51 inhibitors against T. cruzi by applying systematic review strategies, risk of bias assessment, and meta-analysis from in vivo studies. PubMed and Embase databases were searched for original articles, obtaining fifty-six relevant papers meeting inclusion criteria. Characteristics of animal models, parasite strain, treatment schemes, and cure rates were extracted. Primary outcomes such as maximum parasitaemia values, survival, and parasitological cure were recorded for meta-analysis, when possible. The risk of bias was uncertain in most studies. Animals treated with itraconazole, RAV, or POS survived significantly longer than the infected non-treated groups (RR = 4.85 [3.62, 6.49], P < 0.00001), and they showed no differences with animals treated with positive control drugs (RR = 1.01 [0.98, 1.04], P = 0.54). Furthermore, the overall analysis showed that RAV or POS was not likely to achieve parasitological cure when compared with BZ or NFX treatment (OD = 0.49 [0.31, 0.77], P = 0.002). This systematic review contributes to understanding why the azoles had failed in clinical trials and, more importantly, how to improve the animal models of T. cruzi infection by filling the gaps between basic, translational, and clinical research.

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Introduction: Shift work has been hypothesized as a potential risk factor for overweight/obesity or other metabolic changes. We examined the relationship between work shift and body mass index, waist-hip ratio, lipid profile, and glucose concentration in workers from a food manufacturing factory in Colombia. Objectives: To investigate the association between shift work and changes in physiological variables in food manufacturing industry workers in Medellín, Colombia. Methods: This cross-sectional study was conducted with 763 employees from a food manufacturing factory. Information was collected from the medical records from the occupational health provider institution in charge of workers' periodic follow-up. Results: The study sample consisted of 637 (83.5%) men and 126 (16.5%) women. Mean age was 43.35 ± 9.8 years, and mean body mass index was 25.49 ± 3.23 kg/m2. After adjusting for potential confounders, logistic multivariate regression revealed a statistically significant association between shift work and higher body mass index and higher total cholesterol levels compared with dayshift (p < 0.05). Finally, the analysis of waist-hip ratio for each shift scheme and sex showed that this ratio was higher for rotating shift workers, with a significant difference for women. Conclusions: Significant associations were observed between shift work and overweight/obesity and hypercholesterolemia. However, these findings should be confirmed by longitudinal studies.

Introducción: El trabajo por turnos se ha asociado como un potencial factor de riesgo para sobrepeso/obesidad u otras alteraciones metabólicas. Examinamos la relación entre el trabajo por turnos y el índice de masa corporal, índice cintura-cadera, perfil lipídico y glicemia en trabajadores de la industria manufacturera de alimentos en Colombia. Objetivos: Conocer la asociación entre el trabajo por turnos y alteraciones en variables fisiológicos en trabajadores del sector de alimentos en Medellín, Colombia. Métodos: Este estudio transversal fue realizado con 763 trabajadores de una planta del sector de la industria alimentaria. La información fue analizada a partir de registros médicos en la base de datos de una institución prestadora de servicios de salud ocupacional encargada del seguimiento periódico de los trabajadores. Resultados: La muestra del estudio estuvo compuesta por 637 (83,5%) hombres y 126 (16,5%) mujeres. La edad media fue de 43,35 ± 9,8 años y el índice de masa corporal medio de 25,49 ± 3,23 kg/m2. Después de ajustar los posibles factores de confusión, la regresión logística multivariada reveló una asociación estadísticamente significativa entre el trabajo por turno y un mayor índice de masa corporal y nivel de colesterol total, comparado con el turno diurno (p < 0,05). Finalmente, el análisis de la relación cintura-cadera para cada esquema de turno y sexo mostro que esta medida fue mayor para los trabajadores en turno rotativo, siendo significativa la diferencia para las mujeres. Conclusiones: Se observaron asociaciones significativas entre el trabajo por turnos y el sobrepeso/obesidad e hipercolesterolemia; sin embargo, estos hallazgos deben confirmarse mediante estudios longitudinales.

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The discovery and improvements of antimicrobial peptides (AMPs) have become an alternative to conventional antibiotics. They are usually small and heat-stable peptides, exhibiting inhibitory activity against Gram-negative and Gram-positive bacteria. In this way, studies on broad-spectrum AMPs found in amphibians with the remarkable capability to regenerate a wide array of tissues are of particular interest in the search for new strategies to treat multidrug-resistant bacterial strains. In this work, the use of bioinformatic approaches such as sequence alignment with Fasta36 and prediction of antimicrobial activity allowed the identification of the Ramosin peptide from the de novo assembled transcriptome of the plethodontid salamander Bolitoglossa ramosi obtained from post-amputation of the upper limb tissue, heart, and intestine samples. BLAST analysis revealed that the Ramosin peptide sequence is unique in Bolitoglossa ramosi. The peptide was chemically synthesized, and physicochemical properties were characterized. Furthermore, the in vitro antimicrobial activity against relevant Gram-positive and Gram-negative human pathogenic bacteria was demonstrated. Finally, no effect against eukaryotic cells or human red blood cells was evidenced. This is the first antibacterial peptide identified from a Colombian endemic salamander with interesting antimicrobial properties and no hemolytic activity.

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OBJECTIVES: We aimed to contribute to the understanding of the ancient geographic origins of the uniparentally inherited markers in modern admixed Argentinian populations from central Patagonia with new information provided for the city of Trelew. We attempted to highlight the importance of combining different genetic markers when studying population history. METHODS: The mtDNA control region sequence was typified in 89 individuals and 12 Y-STR and 15 Y-SNP loci were analyzed in 66 males. With these data, analysis of molecular variance and Network analyses were carried out. We exhaustively compared the modern data with ancient mtDNA information. Finally, we tested the differences in continental origins estimated by uniparental and previously published biparental markers. RESULTS: Native American mtDNAs (53.9%) increased when maternal ancestors were born in the northern (81.8%) and southern (58.5%) regions of Argentina or in Chile (77.8%). Population substructure was only observed for Y-chromosome haplotypes. Some mtDNA haplogroups have been present in the area for at least ca. 2762-2430 and ca. 500 (D1g and D1g4 haplogroups) and ca. 6736 and ca. 6620 (C1b and C1c haplogroups) years, respectively. In contrast, haplogroups B2i2 and C1b13, frequent in modern Patagonia populations, had not been found in previous ancient DNA studies of the region. CONCLUSIONS: The results suggest that Native American ancestry is well preserved in the region. Trelew samples had characteristic native mtDNA haplogroups previously described in Chilean and Argentine Patagonian populations, but not observed in ancient samples until now. These findings support the idea that these lineages have a recent regional origin. Finally, the estimated proportions of continental ancestry depend on the genetic marker analyzed.

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BACKGROUND: Assessment of therapeutic response with standard serological diagnostic assays in patients with chronic Chagas disease is a major challenge due to the long persistence of parasite-specific antibodies. The current consensus for parasitological cure is to monitor conversion from positive to negative Trypanosoma cruzi serology (seroreversion). However, because of robust humoral immune response, seroreversion by standard serological tests can take years to decades. Developing novel tests of parasitological cure or surrogates is thus a priority in the Chagas disease field. We aimed to evaluate the MultiCruzi assay as a predictive tool for parasitological cure in a cohort of treated infants and children with acute and chronic Chagas disease enrolled in a long-term retrospective longitudinal study with clinical, serological, and parasitological follow-up, and to explore whether MultiCruzi could predict parasitological cure more quickly than the current reference method. METHODS: Patients from two retrospective paediatric Chagas disease cohort studies with clinical, serological, and parasitological follow-up, diagnosed and treated at the parasitology service, Hospital de Niños Ricardo Gutierrez (Buenos Aires, Argentina) were included in this retrospective cohort study. Serum samples were collected every 6 months to 12 months between Oct 22, 1990, and June 3, 2019, for cohort 1 and 1 month after birth for cohort 2 and then every 3 months for a year between July 23, 2012, and April 19, 2016. We evaluated serological follow-up with the Chagatest ELISA (Wiener Lab, Rosario, Argentina) and used this as a clinical reference method for the evaluation of seroreversion. We compared Chagatest ELISA results with results of MultiCruzi (InfYnity Biomarkers, Lyon, France), a novel antibody profiling multiplex assay, investigating seroreversion events with both of the assays and prediction of seroreversion with MultiCruzi using an interpretation formula. FINDINGS: Combining experimental data from discrete analysis of 15 T cruzi antigens efficiently predicted seroreversion at an early stage, which was later confirmed by conventional T cruzi serology. In cohort 1 (n=69), which included children of three different age groups, we observed differences 2 years after therapy. In the 27 individuals from cohort 1 who were treated within the first 12 months of age, MultiCruzi predicted early seroreversion in 21 (78%) patients whereas nine (33%) patients showed seroreversion with Chagatest ELISA (seroreversion difference 0·44, 95% CI 0·26-0·63; p=0·0005). In the 12 patients from cohort 1 treated between 1 year and 2 years of age, MultiCruzi predicted early seroreversion in six (50%) patients, whereas only one (8%) patient was confirmed to be seronegative with Chagatest ELISA (seroreversion difference 0·42, 95% CI 0·14-0·70; p=0·0253). In the 30 patients from cohort 1 who were treated between 2 years and 19 years of age, MultiCruzi predicted early seroreversion in five (6%) patients, whereas no patients were found to be seronegative with Chagatest ELISA (seroreversion difference 0·17, 0·03-0·30; p=0·0253). In cohort 2 (n=27), which included only children younger than 1 year of age and had a shorter follow up (between 5 months and 32 months), the proportion of reported events was significantly different 180 days after treatment for the T cruzi-positive group (early seroreversion predicted in nine [90%] of ten patients with MultiCruzi and confirmed seroreversion in four [40%] of ten patients with Chagatest ELISA; seroreversion difference 0·50, 95% CI 0·19-0·81; p=0·0253) and for the T cruzi-negative group 90 days (early seroreversion predicted in five [29%] of 17 patients with MultiCruzi and confirmed seroreversion in one [6%] of 17 patients with Chagatest ELISA; seroreversion difference 0·24, 0·03-0·44; p=0·0455) and 180 days (early seroreversion predicted in 17 [100%] of 17 patients with MultiCruzi and confirmed seroreversion only in seven [41%] of 17 patients with Chagatest ELISA; seroreversion difference 0·59, 0·35-0·82; p=0·0016) after treatment. INTERPRETATION: The MultiCruzi assay can be used as a predictive monitoring tool to assess parasitological cure in children. This approach might be a solution to forecast forthcoming seroreversion in treated adults infected with T cruzi, but this requires further investigation. FUNDING: Drugs for Neglected Diseases initiative. TRANSLATIONS: For the Spanish, Portuguese and French translations of the abstract see Supplementary Materials section.

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In order to combat bacterial and cancer resistance, we identified peptides (pugnins) with dual antibacterial l-anticancer activity from the Boana pugnax (B. pugnax) skin transcriptome through in silico analysis. Pugnins A and B were selected owing to their high similarity to the DS4.3 peptide, which served as a template for their alignment to the B. pugnax transcriptome, as well as their function as part of a voltage-dependent potassium channel protein. The secondary peptide structure stability in aqueous medium was evaluated as well, and after interaction with the Escherichia coli (E. coli) membrane model using molecular dynamics. These pugnins were synthesized via solid-phase synthesis strategy and verified by Reverse phase high-performance liquid chromatography (RP-HPLC) and mass spectrometry. Subsequently, their alpha-helix structure was determined by circular dichroism, after which antibacterial tests were then performed to evaluate their antimicrobial activity. Cytotoxicity tests against cancer cells also showed selectivity of pugnin A toward breast cancer (MFC7) cells, and pugnin B toward prostate cancer (PC3) cells. Alternatively, flow cytometry revealed necrotic cell damage with a major cytotoxic effect on human keratinocytes (HaCaT) control cells. Therefore, the pugnins found in the transcriptome of B. pugnax present dual antibacterial-anticancer activity with reduced selectivity to normal eukaryotic cells.

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Haplogroup Q originated in Eurasia around 30,000 years ago. It is present in Y-chromosomes from Asia and Europe at rather low frequencies. Since America is undoubtedly one of the continents where this haplogroup is highly represented, it has been defined as one of the founding haplogroups. Its M3 clade has been early described as the most frequent, with pan-American representation. However, it was also possible to find several other haplogroup Q clades at low frequencies. Numerous mutations have been described for haplogroup Q, allowing analysis of its variability and assignment of its geographic origin. We have analyzed 442 samples of unrelated men from Argentina and Paraguay belonging to haplogroup Q; here we report specifically on 27 Q (xM3) lineages. We tested 3 single-nucleotide polymorphisms (SNPs) by amplified product-length polymorphism (APLP) analysis, 3 SNPs for restriction fragment length polymorphism (RFLP) analysis, 15 SNPs by Sanger sequencing, and 17 short tandem repeats (STRs). Our approach allowed us to identify five subhaplogroups. Q-M3 and Q-CTS2730/Z780 are undoubtedly autochthonous lineages and represent the most frequent subhaplogroups, with significant representation in self-defined aboriginal populations, and their autochthonous status has been previously described. The aim of present work was to identify the continental origin of the remaining Q lineages. Thus, we analyzed the STR haplotypes for the samples and compared them with haplotypes described by other authors for the rest of the world. Even when haplogroup Q lineages have been extensively studied in America, some of them could have their origin in post-Columbian human migration from Europe and Middle East.

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Research on alternatives to the use of animal models and cell cultures has led to the creation of organ-on-a-chip systems, in which organs and their physiological reactions to the presence of external stimuli are simulated. These systems could even replace the use of human beings as subjects for the study of drugs in clinical phases and have an impact on personalized therapies. Organ-on-a-chip technology present higher potential than traditional cell cultures for an appropriate prediction of functional impairments, appearance of adverse effects, the pharmacokinetic and toxicological profile and the efficacy of a drug. This potential is given by the possibility of placing different cell lines in a three-dimensional-arranged polymer piece and simulating and controlling specific conditions. Thus, the normal functioning of an organ, tissue, barrier, or physiological phenomenon can be simulated, as well as the interrelation between different systems. Furthermore, this alternative allows the study of physiological and pathophysiological processes. Its design combines different disciplines such as materials engineering, cell cultures, microfluidics and physiology, among others. This work presents the main considerations of OoC systems, the materials, methods and cell lines used for their design, and the conditions required for their proper functioning. Examples of applications and main challenges for the development of more robust systems are shown. This non-systematic review is intended to be a reference framework that facilitates research focused on the development of new OoC systems, as well as their use as alternatives in pharmacological, pharmacokinetic and toxicological studies.

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Similarly to other populations across the Americas, Argentinean populations trace back their genetic ancestry into African, European and Native American ancestors, reflecting a complex demographic history with multiple migration and admixture events in pre- and post-colonial times. However, little is known about the sub-continental origins of these three main ancestries. We present new high-throughput genotyping data for 87 admixed individuals across Argentina. This data was combined to previously published data for admixed individuals in the region and then compared to different reference panels specifically built to perform population structure analyses at a sub-continental level. Concerning the Native American ancestry, we could identify four Native American components segregating in modern Argentinean populations. Three of them are also found in modern South American populations and are specifically represented in Central Andes, Central Chile/Patagonia, and Subtropical and Tropical Forests geographic areas. The fourth component might be specific to the Central Western region of Argentina, and it is not well represented in any genomic data from the literature. As for the European and African ancestries, we confirmed previous results about origins from Southern Europe, Western and Central Western Africa, and we provide evidences for the presence of Northern European and Eastern African ancestries.

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