Inflammatory recruitment of healthy hematopoietic stem and progenitor cells in the acute myeloid leukemia niche.

Inflammation in the bone marrow (BM) microenvironment is a constitutive component of leukemogenesis in acute myeloid leukemia (AML). Current evidence suggests that both leukemic blasts and stroma secrete proinflammatory factors that actively suppress the function of healthy hematopoietic stem and progenitor cells (HSPCs). HSPCs are also cellular components of the innate immune system, and we reasoned that they may actively propagate the inflammation in the leukemic niche. In two separate congenic models of AML we confirm by evaluation of the BM plasma secretome and HSPC-selective single-cell RNA sequencing (scRNA-Seq) that multipotent progenitors and long-lived stem cells adopt inflammatory gene expression programs, even at low leukemic infiltration of the BM. In particular, we observe interferon gamma (IFN-γ) pathway activation, along with secretion of its chemokine target, CXCL10. We show that AML-derived nanometer-sized extracellular vesicles (EVAML) are sufficient to trigger this inflammatory HSPC response, both in vitro and in vivo. Altogether, our studies indicate that HSPCs are an unrecognized component of the inflammatory adaptation of the BM by leukemic cells. The pro-inflammatory conversion and long-lived presence of HSPCs in the BM along with their regenerative re-expansion during remission may impact clonal selection and disease evolution.

Hemorragia digestiva baja: factores de riesgo de gravedad, necesidad de cirugía de urgencia y mortalidad hospitalaria / Lower gastrointestinal bleeding: risk factors of severity, need for emergency surgery and in-hospital Mortality

RESUMEN Antecedentes: Últimamente creció el interés en poder determinar, en etapas tempranas de las hemorragias digestivas bajas (HDB), aquellos factores de riesgo relacionados con la posibilidad de presentar resultados evolutivos adversos. Objectivo: Determinar los factores de riesgo asociados a sangrados graves, cirugía de urgencia y mortalidad hospitalaria. Material y métodos: Realizamos un estudio observacional y retrospectivo sobre 1.850 pacientes, atendidos en forma consecutiva entre enero de 1999 y diciembre de 2018 por HDB. Para evaluar el riesgo de gravedad analizamos trece variables en las primeras cuatro horas desde la admisión. Para determinar los factores relacionados con la cirugía de urgencia, agregamos la enfermedad diverticular y, para evaluar mortalidad, la cirugía de urgencia y el puntaje (score) preoperatorio de la Sociedad Americana de Anestesiología (ASA). Resultados: De los 1.850 casos, 194 fueron graves y 1656 leves/moderados. Resultaron estadísticamente significativos como factores de mayor gravedad: > 70 años, FC > 120 lat/min., TA < 90 mm Hg, oliguria, hematoquecia masiva, hematocrito < 30%, hemoglobina < 7 g/% y necesidad transfusional. Resultaron predictores significativos de cirugía de urgencia: > 70 años, anti-coagulación, hipotensión arterial, taquicardia, hemoglobina < 7 g/%, oliguria, transfusiones y hematoquecia masiva. Se construyó una fórmula pronóstica de requerimiento de cirugía (sensibilidad 94%, especificidad 74%, valor predictivo positivo 91% y valor predictivo negativo 81%). AUC: 0,89%. Fueron significativos para mortalidad: > 70 años, anticoagulados, hematoquecia masiva, transfusiones y cirugía urgente. De los dieciséis pacientes operados y fallecidos de la serie, quince presentaban un ASA ≥ IV. Conclusiones: Las variables utilizadas resultaron simples, fiables y estadísticamente significativas para predecir gravedad, cirugía de urgencia y mortalidad.

ABSTRACT Background: Background: There has been a growing interest in determining those risk factors associated with adverse outcomes in early stages of lower gastrointestinal bleeding (LGIB). Objective: The aim of our study was to analyze the risk factors associated with severe bleeding, emergency surgery and in-hospital mortality. Material and methods: We conducted an observational and retrospective study on 1850 patients consecutive managed between January 1999 and December 2018 for LGIB. We analyzed thirteen variables within the first four hours of hospitalization to evaluate risk severity. Diverticular disease was considered to determine factors associated with emergency surgery, and the preoperative American Society of Anesthesiologists (ASA) score was used to assess mortality and emergency surgery. Results: Out of 1850 cases, 194 were severe and 1656 were mild/moderate, Patients > 70 years, with HR > 120 beats/min, BP < 90 mm Hg, oliguria, massive hematochezia, hematocrit < 30%, hemoglobin < 7 g% and need for transfusions presented statistically significant associations with severe bleeding. Age > 70 years, anticoagulation, hypotension, tachycardia, hemoglobin < 7 g%, oliguria, need for transfusion and massive hematochezia were significant predictors of emergency surgery. A prognostic formula was constructed to predict the need for surgery (sensitivity 94%, specificity 74%, positive predictive value 91% and negative predictive value 81%). AUC-ROC: 0,89%. Age > 70 years, anticoagulation, massive hematochezia transfusions and emergency surgery were identified as predictors of mortality. Fifteen of the sixteen patients who underwent surgery and died had ASA ≥ grade 4. Conclusions: The variables analyzed are simple, reliable and statistically significant to estimate the risk of severe bleeding, need for emergency surgery and mortality.

Control de daños en las emergencias abdominales no traumáticas: causas, indicaciones, factores de riesgo y resultados / Damage control in non-traumatic abdominal emergencies: causes, indications, risk factors and results

RESUMEN Antecedentes : la estrategia de control de daños (ECD) es muy utilizada para el tratamiento de las emergencias abdominales no traumáticas. Objetivo : describir las causas y condiciones fisiopatológicas del empleo de la ECD, determinar la mortalidad según la etiología, criterios de aplicación y factores de riesgo asociados, y comparar la mortalidad observada con la esperada en una serie consecutiva. Material y métodos : se realizó un estudio observacional y retrospectivo, sobre 118 pacientes tratados con ECD, portadores de peritonitis secundarias y hemorragias abdominales graves, con síndrome compartimental abdominal, sepsis abdominal o sistémica o ambas, hipotensión y parámetros de acidosis metabólica asociados. Se analizaron varios factores de riesgo y se comparó la mortalidad observada versus la esperada (APACHE II). Resultados : 112 pacientes presentaron peritonitis generalizada y 6, sangrados intraabdominales graves. La mortalidad fue mayor en la isquemia intestinal grave (p = 0,002). Estuvo relacionada con mayor número de criterios fisiopatológicos de aplicación y con algunos factores de riesgo: glóbulos blancos (GB) ≥ 10 000 ×mm3, hemoglobina (HB) ≤ 9 g/%, creatininemia ≥ 1,3 mg/%, pH ≤ 7,25, ácido láctico ≥ 2,5 mmol/L, diabetes, puntuación (score) ASA ≥ 4, ≥ 4 operaciones y ausencia de cierre parietal inicial. La mortalidad global observada fue 43,1% y la esperada ‒según APACHE II‒ fue del 53%. Conclusiones : la mortalidad fue significativamente mayor en la isquemia intestinal grave y con la presencia de algunos de los factores de riesgo evaluados. Estuvo asociada al número de criterios de aplicación. La mortalidad observada fue menor que la esperada, aunque no significativa.

ABSTRACT Background : Damage control strategy (DCS) is usually used for the treatment of non-traumatic abdominal emergencies. Objective : The aim of the present study was to describe the main causes and pathophysiologic conditions to perform this strategy, the criteria applied and the associated factors and to compare the observed mortality with the expected mortality in the series. Material and methods : We conducted an observational and retrospective study of 118 patients treated with DSC, with secondary peritonitis and severe abdominal bleeding, abdominal compartment syndrome, abdominal or systemic sepsis or both, hypotension and parameters of metabolic acidosis. Several risk factors were analyzed and it was compared observed versus expected mortality (APACHE II). Results : 112 patients presented generalized peritonitis and 6 had severe intra-abdominal bleeding. Mortality was greater in severe mesenteric ischemia (p = 0.002) and was associated with the number of pathophysiologic criteria used for implementation and with white blood cell (WBC) count ≥ 10 000 x mm3, hemoglobin (Hb) ≤ 9 g/dL, creatinine level ≥ 1.3 mg/dL, pH ≤ 7.25, lactic acid ≥ 2.5 mmol/L, diabetes, ASA score ≥ 4, ≥ 4 operations and open abdomen. The overall observed mortality and expected mortality according to the APACHE II score were 43.1% and 53%, respectively. Conclusions : Mortality was significantly greater in patients with severe mesenteric ischemia, presence of some of the risk factors evaluated and was associated with the number of criteria used for implementation. The observed mortality was non-significantly lower than expected.

AMITE: A Novel Polynomial Expansion for Analyzing Neural Network Nonlinearities.

Polynomial expansions are important in the analysis of neural network nonlinearities. They have been applied thereto addressing well-known difficulties in verification, explainability, and security. Existing approaches span classical Taylor and Chebyshev methods, asymptotics, and many numerical approaches. We find that, while these have useful properties individually, such as exact error formulas, adjustable domain, and robustness to undefined derivatives, there are no approaches that provide a consistent method, yielding an expansion with all these properties. To address this, we develop an analytically modified integral transform expansion (AMITE), a novel expansion via integral transforms modified using derived criteria for convergence. We show the general expansion and then demonstrate an application for two popular activation functions: hyperbolic tangent and rectified linear units. Compared with existing expansions (i.e., Chebyshev, Taylor, and numerical) employed to this end, AMITE is the first to provide six previously mutually exclusive desired expansion properties, such as exact formulas for the coefficients and exact expansion errors. We demonstrate the effectiveness of AMITE in two case studies. First, a multivariate polynomial form is efficiently extracted from a single hidden layer black-box multilayer perceptron (MLP) to facilitate equivalence testing from noisy stimulus-response pairs. Second, a variety of feedforward neural network (FFNN) architectures having between three and seven layers are range bounded using Taylor models improved by the AMITE polynomials and error formulas. AMITE presents a new dimension of expansion methods suitable for the analysis/approximation of nonlinearities in neural networks, opening new directions and opportunities for the theoretical analysis and systematic testing of neural networks.

COVID-19 y cirugía. Impacto en los volúmenes de atención, complicaciones posoperatorias y mortalidad. Resultados después de un año de pandemia / COVID-19 and surgery. Impact on volumes of care, postoperative complications and mortality. Results after one year of the pandemic

RESUMEN Antecedentes: la pandemia por COVID-19 generó importantes cambios en la atención y tratamiento de los pacientes quirúrgicos. Objetivo: los objetivos de este estudio fueron comparar los volúmenes de prestaciones realizadas durante un año de pandemia con un período igual sin pandemia, proyectar su impacto asistencial e institucional, y comparar pacientes COVID+ versus COVID- para determinar complicaciones posoperatorias, mortalidad y los factores de riesgo asociados a estos eventos. Material y métodos: estudio observacional y retrospectivo. Comparamos el volumen de prestaciones realizadas entre el 19/3/20 y el 18/3/21 con idéntico período de 2019/20. Efectuamos un estudio de cohorte emparejada (2:1) entre los pacientes con COVID-19 y sin él y se analizaron las complicaciones posoperatorias, la mortalidad, y doce variables objetivas como factores de riesgo asociados. Resultados: todas las variables prestacionales analizadas disminuyeron, pero solo las internaciones programadas y las cirugías y endoscopias no urgentes cayeron significativamente. De los 979 ingresos, 41 casos fueron COVID+ (4,1%). La mortalidad fue del 29,2% en COVID+ (12/41) vs. 7,3% en COVID- (6/82) P = 0,021. Los factores de riesgo significativos asociados a mortalidad fueron: edad ≥ 75 años, hombres, COVID+, urgencias, neumonía, requerimiento de UTI y ARM. Los pacientes operados presentaron una tasa significativamente mayor de neumonías. El análisis de regresión logística (COVID+ vs. -) mostró que por ser COVID+ y registrar la necesidad de ARM, como variables determinantes, en los COVID+ solo la ARM fue determinante en la mortalidad. Conclusión: la pandemia por COVID-19 disminuyó la actividad prestacional y aumentó la mortalidad de los afectados por la virosis.

ABSTRACT Background: The COVID-19 pandemic produced significant changes in the care and treatment of surgical patients. Objectives: The aims of this study were to compare the volume of services provided during a year of pandemic with an equal period without pandemic, estimate its impact on health care and institutional care, and compare COVID-positive versus COVID-negative patients to determine postoperative complications, mortality and risk factors associated with these events. Material and methods: We conducted an observational and retrospective study, comparing the volume of services performed between March 19, 2020, and March 18, 2021, with the same period in 2019/2020. We performed a matched cohort study (in a 2:1 ratio) between patients with and without COVID-19 and analyzed the postoperative complications, mortality, and twelve objective variables as associated risk factors. Results: There was a significant decrease in planned hospitalizations and non-urgent surgeries and endoscopies, while all the other variables showed a non-significant reduction. Of the 979 admissions, 41 corresponded to COVID-positive patients (4.1%). Mortality was 29.2% in COVID-positive patients (12/41) vs. 7.3%% in those COVID negative (p = 0.021). The significant risk factors associated with mortality were age ≥75 years, male sex, COVID+, emergencies, pneumonia, requirement of ICU and MV. Patients operated on had a significantly higher rate of pneumonia. Logistic regression analysis between COVID+ patients and COVID- patients showed that COVID+ and need for MV were predictors of mortality. In COVID+ patients, only MV was a determinant of mortality. Conclusion: The COVID-19 pandemic reduced healthcare services and increased mortality in patients infected with the virus.

SARS-CoV-2 Omicron efficiently infects human airway, but not alveolar epithelium

In late 2021, the highly mutated SARS-CoV-2 Omicron variant emerged, raising concerns about its potential extensive immune evasion, increased transmissibility and pathogenicity. Here, we used organoids of the human airways and alveoli to investigate Omicrons fitness and replicative potential in comparison with earlier SARS-CoV-2 variants. We report that Omicron replicates more rapidly in the airways and has an increased fitness compared to the early 614G variant and Delta. In contrast, Omicron did not replicate productively in human alveolar type 2 cells. Mechanistically, we show that Omicron does not efficiently use TMPRSS2 for entry or spread through cell-cell fusion. Altogether, our data show that Omicron has an altered tropism and protease usage, potentially explaining its higher transmissibility and decreased pathogenicity.

Tracheostomy Decannulation and Disorders of Consciousness Evolution.

BACKGROUND: Tracheostomy is a frequent surgical procedure in subjects with chronic disorders of consciousness (DOC). There is no consensus about safety of tracheostomy decannulation in this population.The aim of our study was to estimate if DOC improvement is a predictor for tracheostomy decannulation. Secondary outcomes include mortality rate and discharge destination. METHODS: We conducted an observational, retrospective, case-control study at a weaning and rehabilitation center (WRC). We included tracheostomized subjects with DOC admitted between August 2015 and December 2017. We matched groups based on the consciousness level at admission assessed withthe coma recovery scale revised (CRS-R). Subjects who were later decannulated formed the cases, while those that remained tracheostomized at the end of follow-up formed the controls. Improvement of DOC was defined as a progress in the categories of the CRS-R. RESULTS: 22 subjects were included in each group. No significant differences were found in clinical and demographic variables, except that controls had longer neurologic injury evolution (65.5 vs 51 days, P = .047), more tracheostomy days at admission to ourinstitution (53 vs 33.5, P = .02), and higher prevalence of neurological comorbidities (12 vs 4, P = .03). Subjects who improved their DOC had more chances of being decannulated (OR 11.28, 95% CI 1.96-123.08). Tracheostomy decannulation could not be achieved in most subjects who did not improve from vegetative state (VS) (OR 0.13, 95% CI 0.02-0.60). 8 subjects, however, could be decannulated in VS, with only one decannulation failure and no deaths. Mortality was higher in controls (0 vs 6, P = .02), especially among VS (0 vs 5, P = .049). No significant differences were found in discharge destination between groups. CONCLUSIONS: Subjects who improve their DOC are more likely to achieve tracheostomy decannulation. Some subjects in VS were decannulated, with lower mortality than those who remained tracheostomized.

Single-Cell Transcriptomics Links Loss of Human Pancreatic ß-Cell Identity to ER Stress.

The maintenance of pancreatic islet architecture is crucial for proper ß-cell function. We previously reported that disruption of human islet integrity could result in altered ß-cell identity. Here we combine ß-cell lineage tracing and single-cell transcriptomics to investigate the mechanisms underlying this process in primary human islet cells. Using drug-induced ER stress and cytoskeleton modification models, we demonstrate that altering the islet structure triggers an unfolding protein response that causes the downregulation of ß-cell maturity genes. Collectively, our findings illustrate the close relationship between endoplasmic reticulum homeostasis and ß-cell phenotype, and strengthen the concept of altered ß-cell identity as a mechanism underlying the loss of functional ß-cell mass.

Neopterin and CXCL-10 in Cerebrospinal Fluid as Potential Biomarkers of Neuroinvasive Dengue and Chikungunya.

Dengue (DENV) and chikungunya viruses (CHIKV) cause severe neurological complications, sometimes undiagnosed. Therefore, the use of more accessible neuroinflammatory biomarkers can be advantageous considering their diagnostic and prognostic potential for aggravated clinical outcomes. In this study, we aimed to evaluate neopterin and C-X-C motif chemokine ligand 10 (CXCL-10) in cerebrospinal fluid (CSF) for the diagnosis of neuroinvasive DENV and CHIKV. We analyzed the CSF of 66 patients with neurological disorders, comprising 12 neuroinvasive DENV/CHIKV, 20 inflammatory control (viral, bacterial, and fungal meningitis, and autoimmune disorders), and 24 noninflammatory control (cerebrovascular disease, dementia, neoplasm). There was no difference between the concentration of CSF neopterin in the neuroinvasive DENV/CHIKV and control groups. However, there was a significant difference in the CXCL-10 level when comparing the neuroinvasive DENV/CHIKV group and the non-inflammatory control (p < 0.05). Furthermore, we found a linear correlation between neopterin and CXCL-10 CSF levels in the three groups. For the DENV/CHIKV neuroinvasive diagnosis, the ROC curve showed the best cut-off values for CSF neopterin at 11.23 nmol/L (sensitivity of 67% and specificity of 63%), and for CSF CXCL-10 at 156.5 pg/mL (91.7% sensitivity and specificity). These results show that CXCL-10 in CSF represents an accurate neuroinflammatory biomarker that may contribute to neuroinvasive DENV/CHIKV diagnosis.

Measurement error due to self-absorption in calibration-free laser-induced breakdown spectroscopy.

Self-absorption of spectral lines is known to lower the performance of analytical measurements via calibration-free laser-induced breakdown spectroscopy. However, the error growth due to this effect is not clearly assessed. Here we propose a method to quantify the measurement error due to self-absorption based on the calculation of the spectral radiance of a plasma in local thermodynamic equilibrium. Validated through spectroscopic measurements for a binary alloy thin film of compositional gradient, the method evidences that measurement performance lowering due to self-absorption depends on the spectral shape of the analytical transition and on the intensity measurement method. Thus, line-integrated intensity measurements of Stark broadened lines enable accurate analysis, even at large optical thickness, if line width and plasma size are precisely known. The error growth due to self-absorption is significantly larger for line shapes dominated by Doppler broadening and for line-center intensity measurements. The findings present a significant advance in compositional measurements via calibration-free laser-induced breakdown spectroscopy, as they enable straightforward selection of most appropriate analytical lines.

IL-10 and IL-12 (P70) Levels Predict the Risk of Covid-19 Progression in Hypertensive Patients: Insights From the BRACE-CORONA Trial.

Background: Cardiovascular comorbidities such as hypertension and inflammatory response dysregulation are associated with worse COVID-19 prognoses. Different cytokines have been proposed to play vital pathophysiological roles in COVID-19 progression, but appropriate prognostic biomarkers remain lacking. We hypothesized that the combination of immunological and clinical variables at admission could predict the clinical progression of COVID-19 in hypertensive patients. Methods: The levels of biomarkers, including C-reactive protein, lymphocytes, monocytes, and a panel of 29 cytokines, were measured in blood samples from 167 hypertensive patients included in the BRACE-CORONA trial. The primary outcome was the highest score during hospitalization on the modified WHO Ordinal Scale for Clinical Improvement. The probability of progression to severe disease was estimated using a logistic regression model that included clinical variables and biomarkers associated significantly with the primary outcome. Results: During hospitalization, 13 (7.8%) patients showed progression to more severe forms of COVID-19, including three deaths. Obesity, diabetes, oxygen saturation, lung involvement on computed tomography examination, the C-reactive protein level, levels of 15 cytokines, and lymphopenia on admission were associated with progression to severe COVID-19. Elevated levels of interleukin-10 and interleukin-12 (p70) combined with two or three of the abovementioned clinical comorbidities were associated strongly with progression to severe COVID-19. The risk of progression to severe disease reached 97.5% in the presence of the five variables included in our model. Conclusions: This study demonstrated that interleukin-10 and interleukin-12 (p70) levels, in combination with clinical variables, at hospital admission are key biomarkers associated with an increased risk of disease progression in hypertensive patients with COVID-19.

The Prognostic Potential of Human Prostate Cancer-Associated Macrophage Subtypes as Revealed by Single-Cell Transcriptomics.

Macrophages in the tumor microenvironment are causally linked with prostate cancer development and progression, yet little is known about their composition in neoplastic human tissue. By performing single cell transcriptomic analysis of human prostate cancer resident macrophages, three distinct populations were identified in the diseased prostate. Unexpectedly, no differences were observed between macrophages isolated from the tumorous and nontumorous portions of the prostatectomy specimens. Markers associated with canonical M1 and M2 macrophage phenotypes were identifiable, however these were not the main factors defining unique subtypes. The genes selectively associated with each macrophage cluster were used to develop a gene signature which was highly associated with both recurrence-free and metastasis-free survival. These results highlight the relevance of tissue-specific macrophage subtypes in the tumor microenvironment for prostate cancer progression and demonstrates the utility of profiling single-cell transcriptomics in human tumor samples as a strategy to design gene classifiers for patient prognostication. IMPLICATIONS: The specific macrophage subtypes present in a diseased human prostate have prognostic value, suggesting that the relative proportions of these populations are related to patient outcome. Understanding the relative contributions of these subtypes will not only inform patient prognostication, but will enable personalized immunotherapeutic strategies to increase beneficial populations or reduce detrimental populations.

Human organoid systems reveal in vitro correlates of fitness for SARS-CoV-2 B.1.1.7

A new phase of the COVID-19 pandemic has started as several SARS-CoV-2 variants are rapidly emerging globally, raising concerns for increased transmissibility. As animal models and traditional in vitro systems may fail to model key aspects of the SARS-CoV-2 replication cycle, representative in vitro systems to assess variants phenotypically are urgently needed. We found that the British variant (clade B.1.1.7), compared to an ancestral SARS-CoV-2 clade B virus, produced higher levels of infectious virus late in infection and had a higher replicative fitness in human airway, alveolar and intestinal organoid models. Our findings unveil human organoids as powerful tools to phenotype viral variants and suggest extended shedding as a correlate of fitness for SARS-CoV-2. One-Sentence SummaryBritish SARS-CoV-2 variant (clade B.1.1.7) infects organoids for extended time and has a higher fitness in vitro.

Trends towards lower azole susceptibility among 200 Candida tropicalis bloodstream isolates from Brazilian medical centres.

OBJECTIVES: Candida tropicalis is one of the three most frequent species causing candidaemia in Latin America. Despite the high prevalence of C. tropicalis in candidaemia cases in Brazil, little is known about the trends in fluconazole susceptibility over time. The objective of this study was to evaluate temporal trends in azole resistance rates among C. tropicalis bloodstream isolates from patients treated in six Brazilian medical centres over a 12-year period. METHODS: We selected 200 C. tropicalis bloodstream isolates from six medical centres in Brazil collected between 2007 and 2018. Species identification was confirmed by MALDI-TOF/MS. Antifungal susceptibility testing for four antifungal agents was performed by the Clinical and Laboratory Standards Institute (CLSI) microbroth method. RESULTS: Overall, rates of non-susceptibility were 4% and 3.5% to fluconazole and voriconazole, respectively. All isolates were susceptible to amphotericin B and only one isolate was resistant to echinocandins. CONCLUSION: Although we failed to demonstrate statistical differences in the rates of azole resistance documented during the period of analysis, trends towards lower susceptibility to fluconazole and voriconazole were shown.

An organoid-derived bronchioalveolar model for SARS-CoV-2 infection of human alveolar type II-like cells.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), which may result in acute respiratory distress syndrome (ARDS), multiorgan failure, and death. The alveolar epithelium is a major target of the virus, but representative models to study virus host interactions in more detail are currently lacking. Here, we describe a human 2D air-liquid interface culture system which was characterized by confocal and electron microscopy and single-cell mRNA expression analysis. In this model, alveolar cells, but also basal cells and rare neuroendocrine cells, are grown from 3D self-renewing fetal lung bud tip organoids. These cultures were readily infected by SARS-CoV-2 with mainly surfactant protein C-positive alveolar type II-like cells being targeted. Consequently, significant viral titers were detected and mRNA expression analysis revealed induction of type I/III interferon response program. Treatment of these cultures with a low dose of interferon lambda 1 reduced viral replication. Hence, these cultures represent an experimental model for SARS-CoV-2 infection and can be applied for drug screens.

Androgen receptor signalling in macrophages promotes TREM-1-mediated prostate cancer cell line migration and invasion.

The androgen receptor (AR) is the master regulator of prostate cancer (PCa) development, and inhibition of AR signalling is the most effective PCa treatment. AR is expressed in PCa cells and also in the PCa-associated stroma, including infiltrating macrophages. Macrophages have a decisive function in PCa initiation and progression, but the role of AR in macrophages remains largely unexplored. Here, we show that AR signalling in the macrophage-like THP-1 cell line supports PCa cell line migration and invasion in culture via increased Triggering Receptor Expressed on Myeloid cells-1 (TREM-1) signalling and expression of its downstream cytokines. Moreover, AR signalling in THP-1 and monocyte-derived macrophages upregulates IL-10 and markers of tissue residency. In conclusion, our data suggest that AR signalling in macrophages may support PCa invasiveness, and blocking this process may constitute one mechanism of anti-androgen therapy.

Exactitud de la punción aspiración con aguja fina en el diagnóstico de cáncer diferenciado de tiroides / Diagnostic accuracy of fine needle aspiration puncture for diagnosing differentiated thyroid cancer

INTRODUCCIÓN: La importancia clínica de los nódulos tiroideos (NT) radica en la necesidad de excluir cáncer de tiroides (7 al 15% de los casos). En Argentina representa el 2,2% y 0,5% de los cánceres que ocurren anualmente en mujeres y hombres respectivamente. La citología por punción aspiración con aguja fina (PAAF) de NT estima el riesgo de malignidad y es la prueba confirmatoria de elección para el diagnóstico, con una especificidad reportada del 94%.OBJETIVO: Determinar la exactitud diagnóstica de los criterios citopatológicos del Sistema Bethesda (SB) obtenidos por PAAF de NT para el diagnóstico de cáncer de tiroides. Establecer la frecuencia de cada categoría del SB en nuestra población. Calcular el porcentaje de malignidad para cada categoría del SB en nuestra población. PACIENTES Y MÉTODOS: Estudio observacional, retrospectivo, de pruebas diagnósticas. Se incluyeron pacientes adultos con NT sospechosos de malignidad por ecografía y PAAF. Se analizaron las historias clínicas de pacientes de ambos sexos intervenidos mediante cirugía tiroidea entre el 1 de enero de 2012 y el 31 de diciembre de 2018 en la Clínica Universitaria Reina Fabiola de la Ciudad de Córdoba. Se evaluó la sensibilidad, especificidad, valores predictivos positivo y negativo de los criterios citopatológicos del SB obtenidos por PAAF de NT tomando como estándar de oro el resultado anatomopatológico. RESULTADOS: Se incluyeron 191 pacientes. De ellos el 82,20% eran de sexo femenino y la media de edad fue 43,45 ± 13,29. Presentaron diagnóstico de cáncer de tiroides 89 pacientes, correspondiendo la totalidad a CDT. Las tasas de malignidad para las categorías del SB II, III, IV y V fueron del 18,5; 31,6; 62,8 y 90,5%, respectivamente. Al analizar la utilidad del SB como prueba de screening encontramos una sensibilidad para detectar malignidad del 84,6%, con una especificidad del 72,6%, un VPP del 76,7%, un VPN del 81,5% y una exactitud diagnóstica global del 78,8%. Este análisis, en puncionesaltamente sospechosas, aumentó la exactitud diagnóstica de la prueba hasta el 85,2%, sensibilidad 76,5%, especificidad 93%, VPP 90,7% y VPN 81,5%. CONCLUSIONES: La exactitud diagnóstica de los criterios citopatológicos del Sistema Bethesda (SB) obtenidos por PAAF de nódulos tiroideos para el diagnóstico de cáncer de tiroides es buena, por lo cual es uno de los métodos recomendados en el algoritmo diagnóstico de cáncer de tiroides. (AU)

INTRODUCTION: The importance of thyroid nodules (NT) lies in the need to exclude thyroid cancer, which occurs in 7 to 15% of cases. In Argentina it represents 2.2% and 0.5% of all cancers that occur annually in women and men respectively. NT fine needle aspiration (PAAF) cytology estimates the risk of malignancy and is the confirmatory test for diagnosis, with a specificity of 94%. OBJECTIVE: To determine the diagnostic accuracy of the cytopathological criteria of the BS obtained by TN FNA for the diagnosis of thyroid cancer. To establish the frequency of each BS category in our population. To alculate the percentage of malignancy for each category of the BS in our population. PATIENTSAND METHODS: Observational, retrospective, diagnostic test study in which adult patients with ultrasound suspiciousformalignancy TN and FNA were included. Data of patients who underwent thyroid surgery between January 1, 2012 and December 31, 2018 at the Reina Fabiola University Clinic in Córdoba City were analyzed. Sensitivity, specificity, positive and negative predictive values of the cytopathological criteria of the BS obtained by FNA were evaluated, taking as a gold standard the pathological result. RESULTS: 191 patients were included. 82.20% were female and the mean age was 43.45 ± 13.29. 89 patients were diagnosed of thyroid cancer, all of them classified as differentiated thyroid carcinoma (DTC). Regarding the percentages of malignancy in the different categories of the Bethesda system, the rates of malignancy for categories II, III, IV and V were 18.5%; 31.6%; 62.8% and 90.5%, respectively. Regarding the performance of the Bethesda system, when analyzing its usefulness as a screening test (analysis 1: category II vs. IV + V + VI) we found a sensitivity to detect malignancy of 84.6%, with a specificity of 72.6% , a PPV of 76.7%, a NPV of 81.5% and a global diagnostic accuracy of 78.8%. This analysis, in highly suspicious punctures (analysis 2: category II vs. V + VI), increased the diagnostic accuracy of the test to 85.2%, sensitivity 76.5%, specificity 93%, PPV 90.7% and NPV 81.5%. CONCLUSIONS: The diagnostic accuracy of the BS used for the cytopathological evaluation of the risk of malignancy in the FNA of thyroid nodules is good, which is why it is one of the recommended methods in the diagnostic algorithm of thyroid cancer. (AU)

Preclinical Investigation of Methylene Blue-mediated Antimicrobial Photodynamic Therapy on Leishmania Parasites Using Real-Time Bioluminescence.

Cutaneous leishmaniasis (CL) is a neglected disease that promotes destructive lesions. Difficulties in treatment are related to accessibility of drugs, resistance and toxicity. Antimicrobial photodynamic therapy (APDT) has been emerging as a promising treatment for CL. In this work, we evaluated methylene blue (MB)-mediated APDT (MB-APDT) on Leishmania amazonensis in vitro and in vivo by bioluminescence technique. In vitro, MB-APDT was performed using a red LED (λ = 660 ± 11 nm, 100 mW cm-2 ) and MB (100 µm) at different light doses. In vivo, mice were infected and 4 weeks later, randomly divided into three groups: control, APDT 1 (single session) and APDT 2 (two sessions of MB-APDT). MB was used at 100 µm and energy dose was established at 150 J cm-2 . Parasite burden, lesion size and pain were evaluated weekly for 4 weeks. In vitro, lethal dose for 90% parasite inactivation was achieved at 48.8 J cm-2 . In vivo, although APDT 1 and APDT 2 groups have showed similar parasite burden after 4 weeks, two sessions were clinically better, especially considering the inflammatory process associated to CL. Our findings reinforce MB-APDT as a cost-effective treatment to combat CL.

Single-cell analysis uncovers that metabolic reprogramming by ErbB2 signaling is essential for cardiomyocyte proliferation in the regenerating heart.

While the heart regenerates poorly in mammals, efficient heart regeneration occurs in zebrafish. Studies in zebrafish have resulted in a model in which preexisting cardiomyocytes dedifferentiate and reinitiate proliferation to replace the lost myocardium. To identify which processes occur in proliferating cardiomyocytes we have used a single-cell RNA-sequencing approach. We uncovered that proliferating border zone cardiomyocytes have very distinct transcriptomes compared to the nonproliferating remote cardiomyocytes and that they resemble embryonic cardiomyocytes. Moreover, these cells have reduced expression of mitochondrial genes and reduced mitochondrial activity, while glycolysis gene expression and glucose uptake are increased, indicative for metabolic reprogramming. Furthermore, we find that the metabolic reprogramming of border zone cardiomyocytes is induced by Nrg1/ErbB2 signaling and is important for their proliferation. This mechanism is conserved in murine hearts in which cardiomyocyte proliferation is induced by activating ErbB2 signaling. Together these results demonstrate that glycolysis regulates cardiomyocyte proliferation during heart regeneration.

Precisión diagnóstica de la ecografía en el diagnóstico de cáncer diferenciado de tiroides / Diagnostic accuracy of ultrasonography for diagnosing differentiated thyroid cancer

INTRODUCCIÓN: El carcinoma diferenciado de tiroides es la neoplasia endócrina más prevalente. En Argentina representa el 2,2% de los cánceres en mujeres y en los hombres corresponde al 0,5%. La ecografía es el método inicial de screening de los nódulos tiroideos y la citología por aspiración con aguja fina (PAAF) es la prueba diagnóstica confirmatoria de elección. Sin embargo, existe una limitación debido a la variabilidad intra e interobservador en los estudios de imágenes y citopatológico. OBJETIVO: Determinar la precisión diagnóstica de los criterios ultrasonográficos sospechosos de malignidad de nódulos tiroideos para el diagnóstico de cáncer de tiroides. Determinar el valor de corte del número de criterios ecográficos de sospecha de malignidad que muestre el mejor balance de sensibilidad/especificidad para diagnosticar cáncer de tiroides. PACIENTES Y MÉTODOS: Estudio observacional, retrospectivo, de pruebas diagnósticas. Se incluyeron pacientes adultos con nódulos sospechosos de malignidad por ecografía y punción aspirativa. Se analizaron las historias clínicas de pacientes de ambos sexos intervenidos mediante cirugía tiroidea entre el 1 de enero de 2012 y el 31 de diciembre de 2018 en la Clínica Universitaria Reina Fabiola en la Ciudad de Córdoba. Se evaluó la sensibilidad, especificidad, valores predictivos positivo y negativo de la presencia de al menos un criterio ecográfico positivo para malignidad tomando como gold standard el resultado anatomopatológico. Se analizó el valor de corte del número de criterios ecográficos que mostrara la mejor relación de sensibilidad/especificidad. RESULTADOS: Se incluyeron 223 pacientes (56%). De ellos 179 (80%) eran de sexo femenino, con una edad media (desviación estándar - DE) de 43,49 (13,03) años. Entre los pacientes con patología benigna, 65 casos (52%) resultaron en adenoma folicular (hallazgo más frecuente). Noventa y nueve (44%) pacientes presentaron diagnóstico de carcinoma diferenciado de tiroides (CDT). Setenta y seis (76.81%) eran de sexo femenino con una edad media (DE) de 40,36 (12,48) años. La media (DE) del tamaño tumoral fue 20,35 (12,73) mm. El 94,45% resultaron CDT papilar cuya variante más frecuente fue la clásica con 72 de 94 (73%) casos. La media (DE) de criterios ecográficos sospechosos en el grupo de pacientes con malignidad fue de 2,10 (1,22) y para el grupo de pacientes con resultado benigno fue de 1,36 (1,08) (p <0,001). La sensibilidad de los criterios ecográficos sospechosos de malignidad para el diagnóstico de CDT fue superior en el criterio hipoecogénico con el 70,7%, y la especificidad fue alta en los siguientes criterios: más alto que ancho, microcalcificaciones, bordes irregulares, invasión perinodular y adenopatías. El valor de corte del número de criterios ecográficos sospechosos de malignidad que mostró el mejor balance de sensibilidad/especificidad para diagnosticar cáncer de tiroides. CONCLUSIONES: La ecografía es el método de screening inicial de los nódulos tiroideos. Si bien algunos criterios ecográficos tienen mayor sensibilidad que otros para el diagnóstico de cáncer de tiroides, la precisión diagnóstica del método se incrementa a medida que aumenta el número de características sospechosas. (AU)

INTRODUCTION: Differentiated thyroid carcinoma is the most prevalent endocrine neoplasm. In Argentina, it represents 2.2% of all cancers that occur annually in women while in men it corresponds to 0.5% of the total cases detected. Ultrasound is the initial screening method for thyroid nodules and fine needle aspiration cytology (FNA) is the confirmatory test for the most appropriate diagnosis and treatment. However, there is a limitation due to intra and interobserver variability in imaging and cytopathological studies. OBJECTIVES: To determine the diagnostic accuracy of the ultrasonographic features for the diagnosis od thyroid nodules malignancy. To determine the cut-off value of the number of ultrasound suspicious criteria for malignancy that shows the best balance of sensitivity / specificity for diagnosing thyroid carcinoma. PATIENTS AND METHOD: This is an observational, retrospective, diagnostic test study that included adult patients with ultrasound and aspiration punction of nodules suspicious of malignancy. The sensitivity, specificity, positive and negative predictive values of the presence of at least one positive ultrasound criterion for malignancy were evaluated, taking the histopathology results as a gold standard. Likewise, it analyzed the cut-off value of the number of ultrasound criteria that shows the best sensitivity / specification ratio. RESULTS: Data from 223 patients (56%) were included. Of these, 179 (81%) were female, with a mean age (standard deviation - SD) of 43.49 (13.03) years. Among patients with benign pathology, 65 (52%) cases resulted in follicular adenoma (most frequent pathological finding). Ninety nine (45%) patients had a diagnosis of differentiated thyroid carcinoma. Seventy six (77%) were female with a mean age (SD) of 40.36 (12.48) years. The mean (SD) of the tumor size was 20.35 (13) mm. Ninety five percent were papillary CDT whose most frequent variant was the classic one with 72 of 94 (73%) cases. The mean (SD) of suspicious ultrasound criteria in the group of patients with malignancy was 2.10 (1.22), for the group of patients with benign results it was 1.36 (1.08) (p <0.001). The sensitivity of the ultrasonographic criteria suspected for malignancy for the diagnosis of differentiated thyroid cancer was higher for the hypoechogenic criterion (0.7) and the specificity was higher in the following criteria: taller than wide, microcalcifications, irregular borders, perinodular invasion and adenopathies. The cut-off value of the number of ultrasonographic criteria suspicious of malignancy that showed the best sensitivity / specificity balance for diagnosing thyroid cancer calculated was 2. CONCLUSION: Ultrasound is the initial screening method for thyroid nodules. Although some ultrasound criteria are more sensitive for the diagnosis of thyroid cancer than others, the diagnostic accuracy of the method increases as the number of suspicious features detected increases. (AU)