Efficacy of short-course treatment for prevention of congenital transmission of Chagas disease: A retrospective cohort study.

BACKGROUND: In regions with controlled vector transmission of T. cruzi, congenital transmission is the most frequent route of infection. Treatment with benznidazole (BZ) or nifurtimox (NF) for 60 days in girls and women of childbearing age showed to be effective in preventing mother to child transmission of this disease. Reports on short-course treatment (≤30 days) are scarce. METHODS: Retrospective cohort study. Offspring of women with Chagas disease who received short-course treatment (≤30 days) with BZ or NF, attended between 2003 and 2022, were evaluated. Parasitemia (microhaematocrit and/or PCR) was performed at <8 months of age, and serology (ELISA and IHA) at ≥8 months to rule out congenital infection. RESULTS: A total of 27 women receiving ≤30 days of treatment and their children were included in this study. NF was prescribed in 17/27 (63%) women, and BZ in 10/27 (37%). The mean duration of treatment was 29.2 days. None of the women experienced serious adverse events during treatment, and no laboratory abnormalities were observed. Forty infants born to these 27 treated women were included. All newborns were full term, with appropriate weight for their gestational age. No perinatal infectious diseases or complications were observed. DISCUSSION: Several studies have shown that treatment of infected girls and women of childbearing age for 60 days is an effective practice to prevent transplacental transmission of T. cruzi. Our study demonstrated that short-duration treatment (≤30 days) is effective and beneficial in preventing transplacental transmission of Chagas disease.

Efficacy and safety of fexinidazole for treatment of chronic indeterminate Chagas disease (FEXI-12): a multicentre, randomised, double-blind, phase 2 trial.

BACKGROUND: More than six million people worldwide, particularly in vulnerable communities in Latin America, are infected with Trypanosoma cruzi, the causative agent of Chagas disease. Only a small portion have access to diagnosis and treatment. Both drugs used to treat this chronic, neglected infection, benznidazole and nifurtimox, were developed more than 50 years ago, and adverse drug reactions during treatment pose a major barrier, causing 20% of patients to discontinue therapy. Fexinidazole proved efficacious in an earlier, interrupted clinical trial, but the doses evaluated were not well tolerated. The present study evaluated fexinidazole at lower doses and for shorter treatment durations. METHODS: In this randomised, double-blind, phase 2 trial, we included adult patients (18-60 years old) with confirmed T cruzi infection by serology and PCR and without signs of organ involvement. We evaluated three regimens of fexinidazole-600 mg once daily for 10 days (6·0 g total dose), 1200 mg daily for 3 days (3·6 g), and 600 mg daily for 3 days followed by 1200 mg daily for 4 days (6·6 g)-and compared them with a historical placebo control group (n=47). The primary endpoint was sustained negative results by PCR at end of treatment and on each visit up to four months of follow-up. This study is registered with ClinicalTrials.gov, NCT03587766, and EudraCT, 2016-004905-15. FINDINGS: Between Oct 16, 2017, and Aug 7, 2018, we enrolled 45 patients (n=15 for each group), of whom 43 completed the study. Eight (19%) of 43 fexinidazole-treated patients reached the primary endpoint, compared with six (13%) of 46 in the historical control group. Mean parasite load decreased sharply following treatment but rebounded beginning 10 weeks after treatment. Five participants had seven grade 3 adverse events: carpal tunnel, sciatica, device infection, pneumonia, staphylococcal infection, and joint and device dislocation. Two participants discontinued treatment due to adverse events unrelated to fexinidazole. INTERPRETATION: The fexinidazole regimens in this study had an acceptable safety profile but did not prove effective against T cruzi infection. Development of fexinidazole monotherapy for treating T cruzi infection has been stopped. FUNDING: The Drugs for Neglected Diseases initiative.

Efficacy of three benznidazole dosing strategies for adults living with chronic Chagas disease (MULTIBENZ): an international, randomised, double-blind, phase 2b trial.

BACKGROUND: Treatment with benznidazole for chronic Chagas disease is associated with low cure rates and substantial toxicity. We aimed to compare the parasitological efficacy and safety of 3 different benznidazole regimens in adult patients with chronic Chagas disease. METHODS: The MULTIBENZ trial was an international, randomised, double-blind, phase 2b trial performed in Argentina, Brazil, Colombia, and Spain. We included participants aged 18 years and older diagnosed with Chagas disease with two different serological tests and detectable T cruzi DNA by qPCR in blood. Previously treated people, pregnant women, and people with severe cardiac forms were excluded. Participants were randomly assigned 1:1:1, using a balanced block randomisation scheme stratified by country, to receive benznidazole at three different doses: 300 mg/day for 60 days (control group), 150 mg/day for 60 days (low dose group), or 400 mg/day for 15 days (short treatment group). The primary outcome was the proportion of patients with a sustained parasitological negativity by qPCR during a follow-up period of 12 months. The primary safety outcome was the proportion of people who permanently discontinued the treatment. Both primary efficacy analysis and primary safety analysis were done in the intention-to-treat population. The trial is registered with EudraCT, 2016-003789-21, and ClinicalTrials.gov, NCT03191162, and is completed. FINDINGS: From April 20, 2017, to Sept 20, 2020, 245 people were enrolled, and 234 were randomly assigned: 78 to the control group, 77 to the low dose group, and 79 to the short treatment group. Sustained parasitological negativity was observed in 42 (54%) of 78 participants in the control group, 47 (61%) of 77 in the low dose group, and 46 (58%) of 79 in the short treatment group. Odds ratios were 1·41 (95% CI 0·69-2·88; p=0·34) when comparing the low dose and control groups and 1·23 (0·61-2·50; p=0·55) when comparing short treatment and control groups. 177 participants (76%) had an adverse event: 62 (79%) in the control group, 56 (73%) in the low dose group, and 59 (77%) in the short treatment group. However, discontinuations were less frequent in the short treatment group compared with the control group (2 [2%] vs 11 [14%]; OR 0·20, 95% CI 0·04-0·95; p=0·044). INTERPRETATION: Participants had a similar parasitological responses. However, reducing the usual treatment from 8 weeks to 2 weeks might maintain the same response while facilitating adherence and increasing treatment coverage. These findings should be confirmed in a phase 3 clinical trial. FUNDING: European Community's 7th Framework Programme.

Follow-up for More than 10 Years of Patients with Peritoneal Metastases Treated with Cytoreductive Surgery + Hyperthermic Intraperitoneal Chemotherapy in a Specialized Unit.

Cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) have demonstrated their impact on disease-free survival (DFS) and overall survival (OS) of patients with peritoneal metastases (PM). However, prior literature lacks evidence regarding any follow-up beyond 5 years. In this study, we analyse long-term OS and DFS (more than 10 years of follow-up) of patients undergoing CRS + HIPEC in a specialized unit. We conducted a retrospective study that included only patients who underwent CRS + HIPEC from January 2001 to May 2012. Data collection was conducted by reviewing medical records and telephone calls to patients or relatives. A total of 86 patients were included. The mean PCI was nine (range 0-39) and complete cytoreduction (CC-0) was reached in 80% of patients. Postoperative complications Clavien-Dindo III-IV occurred in 27.9% of patients and the 30-day mortality rate was 2.3%. After 10 years of actual follow-up, OS was 33.7% and DFS was 31.4%. Considering the historical context in which the standard of care for patients with PM was palliation, the results obtained show that CRS + HIPEC was a valid option, with morbimortality comparable to other major abdominal surgeries and encouraging survival results, since, after 10 years of follow-up, almost one-third of patients are still alive and disease-free.

Venopulmonary Artery Extracorporeal Life Support (VPa ECMO): A Novel Strategy for Refractory Hypoxemia Complicating VV ECMO.

Refractory hypoxemia (RH) during venovenous extracorporeal membrane oxygenation (VV ECMO) support is a complex problem that limits the benefit of this therapy. The need for sustained deep sedation and delays in active rehabilitation are considered as a direct consequence of RH. Changing from VV ECMO to a configuration that returns the flow to pulmonary artery, such as venopulmonary extracorporeal membrane oxygenation (VPa ECMO) may decrease recirculation and improve systemic oxygen delivery. We present a retrospective report that describes the impact of VPa ECMO on oxygenation during sedation withdrawal in 41 patients who received VV ECMO for coronavirus disease 2019 (COVID-19). We evidenced that arterial oxygen pressure (PaO2) increased from 68 to 112.3 mm Hg (p = 0.001) with a reduction of ECMO flow (5.7-4.8 L/m; p = 0.001). Other findings included lower rates of depth sedation (Richmond Agitation Sedation Scale [RASS] ≤3, 37-63%; p = 0.007) and lower requirement inotropic support assessed by LVIS score (4.7-1.1; p = 0.005). Discharge survival was 54% with a sustained benefit until day 79. This cannulation strategy improved effectively PaO2 in this cohort, it may be an alternative in patients with RH in VV ECMO.

A novel, stain-free, natural auto-fluorescent signal, Sig M, identified from cytometric and transcriptomic analysis of infectivity of Cryptosporidium hominis and Cryptosporidium parvum.

Cryptosporidiosis is a worldwide diarrheal disease caused by the protozoan Cryptosporidium. The primary symptom is diarrhea, but patients may exhibit different symptoms based on the species of the Cryptosporidium parasite they are infected with. Furthermore, some genotypes within species are more transmissible and apparently virulent than others. The mechanisms underpinning these differences are not understood, and an effective in vitro system for Cryptosporidium culture would help advance our understanding of these differences. Using COLO-680N cells, we employed flow cytometry and microscopy along with the C. parvum-specific antibody Sporo-Glo™ to characterize infected cells 48 h following an infection with C. parvum or C. hominis. The Cryptosporidium parvum-infected cells showed higher levels of signal using Sporo-Glo™ than C. hominis-infected cells, which was likely because Sporo-Glo™ was generated against C. parvum. We found a subset of cells from infected cultures that expressed a novel, dose-dependent auto-fluorescent signal that was detectable across a range of wavelengths. The population of cells that expressed this signal increased proportionately to the multiplicity of infection. The spectral cytometry results confirmed that the signature of this subset of host cells closely matched that of oocysts present in the infectious ecosystem, pointing to a parasitic origin. Present in both C. parvum and C. hominis cultures, we named this Sig M, and due to its distinct profile in cells from both infections, it could be a better marker for assessing Cryptosporidium infection in COLO-680N cells than Sporo-Glo™. We also noted Sig M's impact on Sporo-Glo™ detection as Sporo-Glo™ uses fluoroscein-isothiocynate, which is detected where Sig M also fluoresces. Lastly, we used NanoString nCounter® analysis to investigate the transcriptomic landscape for the two Cryptosporidium species, assessing the gene expression of 144 host and parasite genes. Despite the host gene expression being at high levels, the levels of putative intracellular Cryptosporidium gene expression were low, with no significant difference from controls, which could be, in part, explained by the abundance of uninfected cells present as determined by both Sporo-Glo™ and Sig M analyses. This study shows for the first time that a natural auto-fluorescent signal, Sig M, linked to Cryptosporidium infection can be detected in infected host cells without any fluorescent labeling strategies and that the COLO-680N cell line and spectral cytometry could be useful tools to advance the understanding of Cryptosporidium infectivity.

Soft skills in personnel training: Report of publications in scopus, topics explored and future research agenda.

Recent research has documented the interest of organizations in training their staff in soft skills, but few studies have been found. Therefore, the objective of this research was to analyze 753 publications in the Scopus database related to soft skills in staff training during the period 1999-2021. These documents were analyzed to identify the main information, the most explored areas, and a future research agenda; all under a bibliometric and bibliographic approach with the help of RStudio and VOSviewer software. The results showed that the keywords with the most co-occurrence were personnel training (n = 110) and soft skills (n = 79). The year with the most documents was 2021 (n = 121). The country with the most publications was the United Kingdom (n = 199). Medicine is the subject area with the most documents (n = 278) and the Article is the type of document with the most studies (n = 566). Eleven areas of further exploration were identified: "Soft skills in software engineering at the higher education level", "Soft skills and communication", "Soft skills and engineering education", "Soft skills in virtual environments", "Soft skills in machine learning", "Serious games in teaching soft skills", "Soft skills for problem-based learning", "Soft skills for project management", "Soft skills and technical skills", "Project-based learning for the assessment of soft skills" and "Soft leadership skills". Five potential areas for future research were derived: soft skills in collaborative work (CSCL), soft skills in computer-aided collaborative work (CSCW), facial expressions as a mirror of soft skills, soft skills for employability and Professional Development Plan (PDP) to assess soft skills. In conclusion, this Review type document on soft skills in personnel training helped to identify the most studied topics during the evaluated period, as well as to identify the little explored topics for future research.

Diagnosis of protozoa diarrhoea in Campylobacter patients increases markedly with molecular techniques.

Cryptosporidium and Giardia are major causes of diarrhoea globally, and two of the most notified infectious diseases in New Zealand. Diagnosis requires laboratory confirmation carried out mostly via antigen or microscopy-based techniques. However, these methods are increasingly being superseded by molecular techniques. Here we investigate the level of protozoa detection by molecular methods in campylobacteriosis cases missed through antigen-based assays and investigate different molecular testing protocols. We report findings from two observational studies; the first among 111 people during a Campylobacter outbreak and the second during normal surveillance activities among 158 people presenting with diarrhoea and a positive Campylobacter test, but negative Cryptosporidium and Giardia antigen-based test results. The molecular methods used for comparison were in-house end-point PCR tests targeting the gp60 gene for Cryptosporidium and gdh gene for Giardia. DNA extraction was performed with and without bead-beating and comparisons with commercial real-time quantitative (qPCR) were made using clinical Cryptosporidium positive sample dilutions down to 10-5. The Cryptosporidium prevalence was 9% (95% CI: 3-15; 10/111) and Giardia prevalence 21% (95% CI: 12-29; 23/111) in the 111 Campylobacter outbreak patients. The Cryptosporidium prevalence was 40% (95% CI: 32-48; 62/158) and Giardia prevalence 1.3% (95% CI: 0.2-4.5; 2/158) in the 158 routine surveillance samples. Sequencing identified Cryptosporidium hominis, C. parvum, and Giardia intestinalis assemblages A and B. We found no statistical difference in positive test results between samples using end-point PCR with or without bead-beating prior to DNA extraction, or between the in-house end-point PCR and qPCR. The qPCR Ct value was 36 (95% CI: 35-37) for 1 oocyst, suggesting a high limit of detection. In conclusion in surveillance and outbreak situations we found diagnostic serology testing underdiagnoses Cryptosporidium and Giardia coinfections in Campylobacter patients, suggesting the impact of protozoa infections may be underestimated through underdiagnosis using antigen-based assays.

Structural and functional integrity of endocrine pancreas post administration of Karwinskia humboldtiana fruit to Wistar rats: a possible therapeutic application for cancer of exocrine origin.

AIMS: Pancreatic adenocarcinoma represents a therapeutic challenge due to the high toxicity of antineoplastic treatments and secondary effects of pancreatectomy. T-514, a toxin isolated from Karwinskia humboldtiana (Kh) has shown antineoplastic activity on cell lines. In acute intoxication with Kh, we reported apoptosis on the exocrine portion of pancreas. One of the mechanisms of antineoplastic agents is the induction of apoptosis, therefore our main objective was to evidence structural and functional integrity of the islets of Langerhans after the administration of Kh fruit in Wistar rats. METHODS: TUNEL assay and immunolabelling against activated caspase-3 were used to detect apoptosis. Also, immunohistochemical tests were performed to search for glucagon and insulin. Serum amylase enzyme activity was also quantified as a molecular marker of pancreatic damage. RESULTS: Evidence of toxicity on the exocrine portion, by positivity in the TUNEL assay and activated caspase-3, was found. On the contrary, the endocrine portion remained structurally and functionally intact, without apoptosis, and presenting positivity in the identification of glucagon and insulin. CONCLUSIONS: These results demonstrated that Kh fruit induces selective toxicity on the exocrine portion and establish a precedent to evaluate T-514 as a potential treatment against pancreatic adenocarcinoma without affecting the islets of Langerhans.

Biomecánica del pase con borde interno entre futbolistas prejuveniles y juveniles de Formativas y Academia / Biomecânica do passe com bordo interior entre os jogadores de futebol pré-juvenil e juvenil de formação e academia / Biomechanics of the pass with internal edge among prejuvenile and juvenile soccer players of Formative and Academy centers

RESUMEN La biomecánica es una ciencia que ayuda al estudio de los movimientos aplicados en distintos deportes, incluyendo el fútbol, que permite identificar y corregir errores técnicos. Esta es una base fundamental para el control de la preparación del deportista. En tal sentido, se planteó como propósito de la investigación analizar biomecánicamente el pase con borde interno del pie en futbolistas prejuveniles y juveniles de Formativas y Academia. La investigación es de tipo descriptiva-explicativa, de orden correlacional; se diagnostican biomecánicamente a 40 futbolistas en dos grupos independientes (grupo 1: Formativas; grupo 2: Academia; entre 14-18 años). Se analizan diez indicadores, se incluye la edad. Se demostraron diferencias significativas en el indicador "DB" (p=0.000), el indicador "ÁFRA" (p=0.006), el indicador "DPAE" (p=0.000), y los indicadores "VF", "A" y "TE" (p=0.000) respectivamente, mientras que las variables o indicadores "ÁFRE" (p=1.000), "EX" (p=0.102) y "EY" (p=0.056) no presentaron diferencias significativas entre los grupos independientes. En la comparación realizada con los jugadores de un equipo profesional y los jugadores de una Academia de formación, se pudo evidenciar que existen diferencias notables desde la fase inicial del pase con el borde interno del pie, mostrando la distancia del jugador con respecto al balón, así como en la fase previa donde los jugadores de Formativas poseen una mejor ejecución del movimiento técnico; esto se refleja en la fase de contacto donde se evidencia una mayor velocidad y aceleración al momento del impacto con el balón.

RESUMO A biomecânica é uma ciência que ajuda o estudo dos movimentos aplicados em diferentes desportos, incluindo o futebol, o que permite a identificação e correção de erros técnicos. Esta é uma base fundamental para o controlo da preparação do atleta. Neste sentido, o objectivo da investigação era analisar biomecanicamente o passe com a extremidade interna do pé em jogadores de futebol pré-juvenil e juvenil em treino e academia. A investigação é de tipo descritivo-explicativo, de ordem correlativa; 40 jogadores de futebol são diagnosticados biomecanicamente em dois grupos independentes (Grupo 1: Formação; Grupo 2: Academia; entre 14-18 anos de idade). Foram analisados dez indicadores, incluindo a idade. Foram demonstradas diferenças significativas no indicador "DB" (p=0,000), o indicador "ÁFRA" (p=0,006), o indicador "DPAE" (p=0,000), e os indicadores "VF", "A" e "TE" (p=0,000) respectivamente, enquanto que as variáveis ou indicadores "ÁFRE" (p=1,000), "EX" (p=0,102) e "EY" (p=0,056) não apresentavam diferenças significativas entre os grupos independentes. Na comparação feita com os jogadores de uma equipa profissional e com os jogadores de uma academia de treino, constatou-se que existem diferenças notáveis em relação à fase inicial do passe com a borda interior do pé, mostrando a distância do jogador em relação à bola, bem como na fase anterior em que os jogadores da academia de treino têm uma melhor execução do movimento técnico; isto reflete-se na fase de contato onde é evidente uma maior velocidade e aceleração no momento do impacto com a bola.

ABSTRACT Biomechanics is a science that helps the study of movements applied in different sports, including soccer, which allows identifying and correcting technical errors. This is a fundamental basis for the control of the athlete's preparation. In this sense, the purpose of the research was to analyze biomechanically the pass with the inner edge of the foot in pre-juvenile and youth soccer players of training and academy. The research is of descriptive-explanatory type, of correlational order; 40 soccer players were biomechanically diagnosed in two independent groups (Group 1: Formative; Group 2: Academy; between 14-18 years). Ten indicators are analyzed, including age. Significant differences were demonstrated in the "DB" indicator (p=0.000), the "ÁFRA" indicator (p=0.006), the "DPAE" indicator (p=0.000), and the "VF", "A" and " TE" (p=0.000) respectively, while the variables or indicators "ÁFRE" (p=1.000), "EX" (p=0.102) and "EY" (p=0.056) did not present significant differences between the independent groups. In the comparison made with the players of a professional team and the players of a training academy, it was possible to show that there are notable differences from the initial phase of the pass with the inner edge of the foot, showing the distance of the player with respect to the ball, as well as in the previous phase where the players in formatiom have a better execution of the technical movement; this is reflected in the contact phase where greater speed and acceleration are evident at the moment of impact with the ball.

Diferencias biomecánicas entre jugadores avanzados y amateur en la ejecución del tiro libre en fútbol / Diferenças biomecânicas entre jogadores avançados e amadores na execução do pontapé livre no futebol / Biomechanical differences between advanced and amateur players in the execution of the free kick in soccer

RESUMEN El tiro libre en el fútbol es una de las técnicas deportivas de importancia, la cual permite reanudar el juego luego de la falta cometida por el otro jugador. La potenciación de la habilidad motriz específica permite desarrollar mayores rendimientos a corto y largo plazo, por lo cual su caracterización es importante para desarrollar metodologías especializadas en las etapas de iniciación deportiva. En tal sentido, la presente investigación tuvo por objetivo analizar las diferencias biomecánicas del tiro libre entre jugadores avanzados y amateur de fútbol. Este estudio es de tipo descriptivo-explicativo de orden correlacional, se diagnostican intencionalmente a 30 futbolistas de la Escuela de Fútbol Chiqui Park (16-32 años de edad), clasificados en dos grupos independientes, el grupo 1 con futbolistas avanzados, y el grupo 2 con futbolistas de nivel amateur. Se evalúan tres variables: el ángulo inicial de la Fase unipodal (AIFU), el ángulo final de la fase final (AFF), y el tiempo de ejecución del tiro libre (TE). No se evidencian diferencias significativas entre grupos independientes en las variables angulares AIFU (p=0.683) y AFF (p=0.389), y una diferencia significativa a favor del grupo 2 en la variable TE (p=0.000). Sin embargo, todos los rangos promedios favorecieron a los futbolistas de nivel avanzado. Existe la necesidad de establecer integralmente mayores estudios que caractericen las categorías formativas estudiadas, sirviendo de base teórica y metodología que fundamente el desarrollo de acciones técnico-tácticas y físicas de consideración en los procesos de dirección del entrenamiento deportivo.

RESUMO O pontapé livre no futebol é uma das técnicas desportivas mais importantes, que permite o reinício do jogo após uma falta cometida pelo outro jogador. A potenciação da habilidade motora específica permite o desenvolvimento de desempenhos mais elevados a curto e longo prazo, razão pela qual a sua caracterização é importante para desenvolver metodologias especializadas nas fases de iniciação ao desporto. Neste sentido, o objectivo desta investigação era analisar as diferenças biomecânicas do pontapé livre entre jogadores de futebol avançados e amadores. Este estudo é de um tipo descritivo-explicativo de ordem correlacional, 30 jogadores de futebol da Escola de Futebol do Chiqui Park (16-32 anos de idade) são intencionalmente diagnosticados, classificados em dois grupos independentes, grupo 1 com jogadores de futebol avançados, e grupo 2 com jogadores de futebol amadores. São avaliados os seguintes São avaliadas três variáveis de interesse: o ângulo inicial da fase unipodal (AIFU), o ângulo final da fase final (AFF), e o tempo de execução do lançamento livre (TE). Não foram evidentes diferenças significativas entre grupos independentes nas variáveis angulares AIFU (p=0,683) e AFF (p=0,389), e uma diferença significativa a favor do grupo 2 na variável TE (p=0,000). No entanto, todas as gamas médias favoreceram os jogadores de nível avançado. Há necessidade de estabelecer de forma abrangente outros estudos que caracterizem as categorias de formação estudadas, servindo como base teórica e metodológica para o desenvolvimento de ações técnico-tácticas e físicas de consideração nos processos de gestão da formação desportiva.

ABSTRACT The free kick in soccer is one of the important sports techniques, which allows the game to resume after a foul committed by the other player. The enhancement of specific motor skills allows the development of higher performance in the short and long term, for which its characterization is important to develop specialized methodologies in the stages of sports initiation. In this sense, the present research aimed to analyze the biomechanical differences of the free kick between advanced and amateur soccer players. This study is of a descriptive-explanatory type of correlational order, 30 soccer players from the Chiqui Park Soccer School (16-32 years of age) are intentionally diagnosed, classified into two independent groups, group 1 with advanced soccer players, and group 2 with amateur level soccerers. Three variables of interest are evaluated: the Initial Angle of the Unipodal Phase (AIFU in Spanish), the Final Angle of the Final Phase (AFF in Spanish), and the Execution Time of the Free Throw (TE in Spanish). There are no significant differences between independent groups in the angular variables AIFU (p=0.683) and AFF (p=0.389), and a significant difference in favor of group 2 in the variable TE (p=0.000). However, all the average ranges favored the advanced level players. There is a need to comprehensively establish larger studies that characterize the training categories studied, serving as a theoretical basis and methodology that supports the development of technical-tactical and physical actions of consideration in the processes of sports training management.

Uncovering the genetic diversity of Giardia intestinalis in isolates from outbreaks in New Zealand.

BACKGROUND: Giardia intestinalis is one of the most common causes of diarrhoea worldwide. Molecular techniques have greatly improved our understanding of the taxonomy and epidemiology of this parasite. Co-infection with mixed (sub-) assemblages has been reported, however, Sanger sequencing is sometimes unable to identify shared subtypes between samples involved in the same epidemiologically linked event, due to samples showing multiple dominant subtypes within the same outbreak. Here, we aimed to use a metabarcoding approach to uncover the genetic diversity within samples from sporadic and outbreak cases of giardiasis to characterise the subtype diversity, and determine if there are common sequences shared by epidemiologically linked cases that are missed by Sanger sequencing. METHODS: We built a database with 1109 unique glutamate dehydrogenase (gdh) locus sequences covering most of the assemblages of G. intestinalis and used gdh metabarcoding to analyse 16 samples from sporadic and outbreak cases of giardiasis that occurred in New Zealand between 2010 and 2018. RESULTS: There is considerable diversity of subtypes of G. intestinalis present in each sample. The utilisation of metabarcoding enabled the identification of shared subtypes between samples from the same outbreak. Multiple variants were identified in 13 of 16 samples, with Assemblage B variants most common, and Assemblages E and A present in mixed infections. CONCLUSIONS: This study showed that G. intestinalis infections in humans are frequently mixed, with multiple subtypes present in each host. Shared sequences among epidemiologically linked cases not identified through Sanger sequencing were detected. Considering the variation in symptoms observed in cases of giardiasis, and the potential link between symptoms and (sub-) assemblages, the frequency of mixed infections could have implications for our understanding of host-pathogen interactions.

Culture of cancer spheroids and evaluation of anti-cancer drugs in 3D-printed miniaturized continuous stirred tank reactors (mCSTRs).

Cancer continues to be a leading cause of mortality in modern societies; therefore, improved and more reliablein vitrocancer models are needed to expedite fundamental research and anti-cancer drug development. Here, we describe the use of a miniaturized continuous stirred tank reactor (mCSTR) to first fabricate and mature cancer spheroids (i.e. derived from MCF7 cells, DU145 cells, and a mix of MCF7 cells and fibroblasts), and then to conduct anti-cancer drug assays under continuous perfusion. This 3 ml mCSTR features an off-center agitation system that enables homogeneous chaotic laminar mixing at low speeds to support cell aggregation. We incubated cell suspensions for 3 d in ultra-low-attachment plates to allow formation of discoid cell aggregates (∼600µm in diameter). These cell aggregates were then transferred into mCSTRs and continuously fed with culture medium. We characterized the spheroid morphology and the expression of relevant tumor biomarkers at different maturation times for up to 4 weeks. The spheroids progressively increased in size during the first 5-6 d of culture to reach a steady diameter between 600 and 800µm. In proof-of-principle experiments, we demonstrated the use of this mCSTR in anti-cancer drug testing. Three drugs commonly used in breast cancer treatment (doxorubicin, docetaxel, and paclitaxel) were probed at different concentrations in MCF7-derived spheroids. In these experiments, we evaluated cell viability, glucose consumption, spheroid morphology, lactate dehydrogenase activity, and the expression of genes associated with drug resistance (ABCB1andABCC1) and anti-apoptosis (Bcl2). We envision the use of this agitated system as a tumor-on-a-chip platform to expedite efficacy and safety testing of novel anti-cancer drugs and possibly in personalized medicine applications.

Genetic polymorphism of Trypanosoma cruzi bloodstream populations in adult chronic indeterminate Chagas disease patients from the E1224 clinical trial.

BACKGROUND: The role that the genetic diversity of natural Trypanosoma cruzi populations plays in response to trypanocidal treatment of chronic Chagas disease (CD) patients remains to be understood. We analysed the genetic polymorphisms of parasite bloodstream populations infecting chronic CD patients enrolled in the E1224 clinical trial. METHODS: A total of 506 baseline and post-treatment follow-up samples from 188 patients were analysed. T. cruzi satellite DNA (satDNA) was amplified and sequenced using cruzi1/cruzi2 primers, and samples with TcI/III, TcII, TcIV or hybrid satDNA sequences were identified. Minicircle signatures were obtained after kinetoplast DNA amplification using 121/122 primers and restriction enzyme digestion. Genetic distances between baseline and post-treatment minicircle signatures were estimated using the Jaccard coefficient. RESULTS: At baseline, 74.3% TcII, 17.9% hybrid and 7.8% TcI/III satDNA sequences were found, whereas at the end of follow-up the distribution was 55.2% TcII, 35.2% hybrid and 9.5% TcI/III. The placebo arm was the treatment group with the highest variation of satDNA sequences between baseline and post-treatment follow-up. Genetic distances between baseline and post-treatment minicircle signatures were similar among all treatment arms. No association between minicircle signature variability and satDNA type distribution was found. CONCLUSIONS: Genetic variability of T. cruzi bloodstream populations during post-treatment follow-up did not differ from that observed during chronic infection in the absence of treatment, suggesting that there were no selection events of E1224-resistant parasite populations. This is the first report documenting the genetic polymorphism of natural T. cruzi populations in chronic patients in the context of clinical trials with trypanocidal drugs.

Abordaje mínimamente invasivo del tumor de Wilms unilateral: un reporte de caso y revisión de la literatura / Minimally invasive approach to unilateral Wilms tumor: a case report and literature review

Introducción. El tumor de Wilms es la neoplasia abdominal más común en pacientes pediátricos. En la mayoría de los casos se presenta como una masa unilateral indolora en el abdomen. El objetivo de este artículo fue presentar el caso de una paciente de 4 años con tumor de Wilms unilateral derecho manejado con cirugía mínimamente invasiva en el Hospital Universitario del Valle. Métodos. Revisión de la historia clínica e imágenes de la paciente, descripción de la técnica quirúrgica y revisión de la literatura del manejo de tumor de Wilms unilateral. Caso clínico. Paciente femenina de 4 años quien consultó en abril de 2019 por un cuadro clínico de dolor abdominal y sensación de masa en flanco derecho. Se realizó ecografía abdominal donde se encontró imagen nodular heterogénea de contornos definidos en riñón derecho, con riñón izquierdo normal. Se hizo diagnóstico de tumor de Wilms unilateral y se llevó a cirugía mediante abordaje mínimamente invasivo, con buena evolución postoperatoria. Conclusión. La cirugía es el pilar del manejo, y la nefrectomía mediante abordaje laparoscópico para casos seleccionados, en manos entrenadas, tiene la suficiente eficacia, seguridad y cumplimiento de los principios quirúrgicos y oncológicos que provee la cirugía abierta.

Introduction. Wilms tumor is the most common abdominal neoplasm in pediatric patients. In most cases it presents as a painless unilateral mass in the abdomen. The objective of this article was to present the case of a 4-year-old patient with right unilateral Wilms tumor managed with minimally invasive surgery at the Hospital Universitario del Valle. Methods. Review of the patient's clinical history and images, description of the surgical technique and review of the literature on the management of unilateral Wilms tumor.Clinical case. A 4-year-old female patient who consulted on April 4, 2019 with a clinical presentation of abdominal pain and sensation of mass in the right flank. Abdominal ultrasound was performed where a heterogeneous nodular image of defined contours was found in the right kidney, with a normal left kidney. A diagnosis of unilateral Wilms tumor was made and surgery was carried out using a minimally invasive approach, with good postoperative evolution. Conclusion. Wilms tumor is the most common abdominal neoplasm in pediatric patients. The most common presentation is a painless palpable mass. Surgery is the mainstay of management, and nephrectomy using a laparoscopic approach for selected cases, in trained hands, has sufficient efficacy, safety, and compliance with surgical and oncological principles that open surgery provides.

Molecular Imaging of Vulnerable Coronary Plaque with Radiolabeled Somatostatin Receptors (SSTR).

Atherosclerosis is responsible for the majority of heart attacks and is characterized by several modifications of the arterial wall including an inflammatory reaction. The silent course of atherosclerosis has made it necessary to develop predictors of disease complications before symptomatic lesions occur. Vulnerable to rupture atherosclerotic plaques are the target for molecular imaging. To this aim, different radiopharmaceuticals for PET/CT have emerged for the identification of high-risk plaques, with high specificity for the identification of the cellular components and pathophysiological status of plaques. By targeting specific receptors on activated macrophages in high-risk plaques, radiolabelled somatostatin analogues such as 68Ga-DOTA-TOC, TATE,0 or NOC have shown high relevance to detect vulnerable, atherosclerotic plaques. This PET radiopharmaceutical has been tested in several pre-clinical and clinical studies, as reviewed here, showing an important correlation with other risk factors.

Manejo quirúrgico de prolapso uretral en pediatría: Reporte de caso / Surgical management of urethral prolapse in pediatrics: A case report

Introducción. El prolapso uretral es una entidad poco común, con una incidencia estimada de 1 en 3000 mujeres. Se presenta cuando la mucosa uretral sobresale espontáneamente más allá del meato uretral. Es una patología poco diagnosticada dada su baja frecuencia y de allí la importancia de conocer sobre su presentación, diagnóstico y tratamiento. El objetivo de este artículo fue presentar el caso de una paciente de 10 años con diagnóstico de prolapso uretral y su manejo quirúrgico. Caso clínico. Paciente femenina de 10 años, que consultó por cuadro clínico de 1 año de evolución consistente en dolor en región urogenital, que se irradiaba a hipogastrio, asociado a pujo y disuria, a quien se le diagnosticó prolapso uretral y se realizó corrección quirúrgica de mucosa uretral prolapsada mediante técnica de Kelly-Burnham modificada. Conclusión. El prolapso uretral es una entidad que con frecuencia es diagnosticada erróneamente pues su diagnóstico es eminentemente clínico. Si bien se ha descrito el tratamiento médico en primera instancia, éste tiene una alta tasa de recurrencia, por lo que en estos casos se prefiere la resección quirúrgica del tejido prolapsado.

Introduction. Urethral prolapse is a rare entity, with an estimated incidence of 1 in 3,000 women. It occurs when the urethral mucosa spontaneously protrudes beyond the urethral meatus. It is a poorly diagnosed pathology given its low frequency and hence the importance of knowing about its presentation, diagnosis and treatment. The objective of this article was to present the case of a 10-year-old patient with a diagnosis of urethral prolapse and its surgical management. Clinical case. A 10-year-old female patient, who consulted for a 1-year clinical picture consisting of pain in the urogenital region, radiating to the hypogastrium, associated with pushing and dysuria, who was diagnosed with urethral prolapse and a surgical correction of the urethral mucosa was performed prolapsed by modified Kelly-Burnham technique. Conclusion. Urethral prolapse is an entity that is frequently misdiagnosed because its diagnosis is eminently clinical. Although medical treatment has been described in the first instance, it has a high recurrence rate, so surgical resection of the prolapsed tissue is preferred in these cases.

Prospective multicenter evaluation of real time PCR Kit prototype for early diagnosis of congenital Chagas disease.

BACKGROUND: Current algorithm for Congenital Chagas Disease (cCD) diagnosis is unsatisfactory due to low sensitivity of the parasitological methods. Moreover, loss to follow-up precludes final serodiagnosis after nine months of life in many cases. A duplex TaqMan qPCR kit for Trypanosoma cruzi DNA amplification was prospectively evaluated in umbilical cord (UCB) and peripheral venous blood (PVB) of infants born to CD mothers at endemic and non-endemic sites of Argentina. METHODS: We enrolled and followed-up 370 infants; qPCR was compared to gold-standard cCD diagnosis following studies of diagnostic accuracy guidelines. FINDINGS: Fourteen infants (3·78%) had cCD. The qPCR sensitivity and specificity were higher in PVB (72·73%, 99·15% respectively) than in UCB (66·67%, 96·3%). Positive and negative predictive values were 80 and 98·73% and 50 and 98·11% for PVB and UCB, respectively. The Areas under the Curve (AUC) of ROC analysis for qPCR and micromethod (MM) were 0·81 and 0·67 in UCB and 0·86 and 0·68 in PVB, respectively. Parasitic loads ranged from 37·5 to 23,709 parasite equivalents/mL. Discrete typing Unit Tc V was identified in five cCD patients and in six other cCD cases no distinction among Tc II, Tc V or Tc VI was achieved. INTERPRETATION: This first prospective field study demonstrated that qPCR was more sensitive than MM for early cCD detection and more accurate in PVB than in UCB. Its use, as an auxiliary diagnostic tool to MM will provide more accurate records on cCD incidence. FUNDING: FITS SALUD 001-CHAGAS (FONARSEC, MINCyT, Argentina) to the Public-Private Consortium (INGEBI-CONICET, INP-ANLIS MALBRAN and Wiener Laboratories); ERANET-LAC-HD 328 to AGS and PICT 2015-0074 (FONCYT, MinCyT) to AGS and FA.

New regimens of benznidazole monotherapy and in combination with fosravuconazole for treatment of Chagas disease (BENDITA): a phase 2, double-blind, randomised trial.

BACKGROUND: Current treatment for Chagas disease with the only available drugs, benznidazole or nifurtimox, has substantial limitations, including long treatment duration and safety and tolerability concerns. We aimed to evaluate the efficacy and safety of new benznidazole monotherapy regimens and combinations with fosravuconazole, in the treatment of Chagas disease. METHODS: We did a double-blind, double-dummy, phase 2, multicentre, randomised trial in three outpatient units in Bolivia. Adults aged 18-50 years with chronic indeterminate Chagas disease, confirmed by serological testing and positive qualitative PCR results, were randomly assigned (1:1:1:1:1:1:1) to one of seven treatment groups using a balanced block randomisation scheme with an interactive response system. Participants were assigned to benznidazole 300 mg daily for 8 weeks, 4 weeks, or 2 weeks, benznidazole 150 mg daily for 4 weeks, benznidazole 150 mg daily for 4 weeks plus fosravuconazole, benznidazole 300 mg once per week for 8 weeks plus fosravuconazole, or placebo, with a 12-month follow-up period. The primary endpoints were sustained parasitological clearance at 6 months, defined as persistent negative qualitative PCR results from end of treatment, and incidence and severity of treatment-emergent adverse events, serious adverse events, and adverse events leading to treatment discontinuation. Primary efficacy analysis was based on the intention-to-treat and per-protocol populations and secondary efficacy analyses on the per-protocol population. Safety analyses were based on the as-treated population. Recruitment is now closed. This trial is registered with ClinicalTrials.gov, NCT03378661. FINDINGS: Between Nov 30, 2016, and July 27, 2017, we screened 518 patients, and 210 were enrolled and randomised. 30 patients (14%) were assigned to each treatment group. All 210 randomised patients were included in the intention-to-treat population, and 190 (90%) were included in the per-protocol population. In the intention-to-treat analysis, only one (3%) of 30 patients in the placebo group had sustained parasitological clearance at 6 months of follow-up. Sustained parasitological clearance at 6 months was observed in 25 (89%) of 28 patients receiving benznidazole 300 mg daily for 8 weeks (rate difference vs placebo 86% [95% CI 73-99]), 25 (89%) of 28 receiving benznidazole 300 mg daily for 4 weeks (86% [73-99]), 24 (83%) of 29 receiving benznidazole 300 mg daily for 2 weeks (79% [64-95]), 25 (83%) of 30 receiving benznidazole 150 mg daily for 4 weeks (80% [65-95]), 23 (85%) of 28 receiving benznidazole 150 mg daily for 4 weeks plus fosravuconazole (82% [67-97]), and 24 (83%) of 29 receiving benznidazole 300 mg weekly for 8 weeks plus fosravuconazole (79% [64-95]; p<0·0001 for all group comparisons with placebo). Six patients (3%) had ten serious adverse events (leukopenia [n=3], neutropenia [n=2], pyrexia, maculopapular rash, acute cholecystitis, biliary polyp, and breast cancer), eight had 12 severe adverse events (defined as interfering substantially with the patient's usual functions; elevated alanine aminotransferase [n=4], elevated gamma-glutamyltransferase [n=2], elevated aspartate aminotransferase [n=1], neutropenia [n=3], leukopenia [n=1], and breast cancer [n=1]), and 15 (7%) had adverse events that led to treatment discontinuation (most of these were in the groups who received benznidazole 300 mg daily for 8 weeks, benznidazole 300 mg once per week for 8 weeks plus fosravuconazole, and benznidazole 150 mg daily for 4 weeks plus fosravuconazole). No adverse events leading to treatment discontinuation were observed in patients treated with benznidazole 300 mg daily for 2 weeks or placebo. There were no treatment-related deaths. INTERPRETATION: Benznidazole induced effective antiparasitic response, regardless of treatment duration, dose, or combination with fosravuconazole, and was well tolerated in adult patients with chronic Chagas disease. Shorter or reduced regimens of benznidazole could substantially improve treatment tolerability and accessibility, but further studies are needed to confirm these results. FUNDING: Drugs for Neglected Diseases initiative (DNDi). TRANSLATION: For the Spanish translation of the abstract see Supplementary Materials section.

Cancer cells educate natural killer cells to a metastasis-promoting cell state.

Natural killer (NK) cells have potent antitumor and antimetastatic activity. It is incompletely understood how cancer cells escape NK cell surveillance. Using ex vivo and in vivo models of metastasis, we establish that keratin-14+ breast cancer cells are vulnerable to NK cells. We then discovered that exposure to cancer cells causes NK cells to lose their cytotoxic ability and promote metastatic outgrowth. Gene expression comparisons revealed that healthy NK cells have an active NK cell molecular phenotype, whereas tumor-exposed (teNK) cells resemble resting NK cells. Receptor-ligand analysis between teNK cells and tumor cells revealed multiple potential targets. We next showed that treatment with antibodies targeting TIGIT, antibodies targeting KLRG1, or small-molecule inhibitors of DNA methyltransferases (DMNT) each reduced colony formation. Combinations of DNMT inhibitors with anti-TIGIT or anti-KLRG1 antibodies further reduced metastatic potential. We propose that NK-directed therapies targeting these pathways would be effective in the adjuvant setting to prevent metastatic recurrence.