Monocyte glycolysis determines CD8+ T cell functionality in human Chagas disease.

Chagas disease is a lifelong pathology resulting from Trypanosoma cruzi infection. It represents one of the most frequent causes of heart failure and sudden death in Latin America. Herein, we provide evidence that aerobic glycolytic pathway activation in monocytes drives nitric oxide (NO) production, triggering tyrosine nitration (TN) on CD8+ T cells and dysfunction in patients with chronic Chagas disease. Monocytes from patients exhibited a higher frequency of hypoxia-inducible factor 1α and increased expression of its target genes/proteins. Nonclassical monocytes are expanded in patients' peripheral blood and represent an important source of NO. Monocytes entail CD8+ T cell surface nitration because both the frequency of nonclassical monocytes and that of NO-producing monocytes positively correlated with the percentage of TN+ lymphocytes. Inhibition of glycolysis in in vitro-infected peripheral blood mononuclear cells decreased the inflammatory properties of monocytes/macrophages, diminishing the frequency of IL-1ß- and NO-producing cells. In agreement, glycolysis inhibition reduced the percentage of TN+CD8+ T cells, improving their functionality. Altogether, these results clearly show that glycolysis governs oxidative stress on monocytes and modulates monocyte-T cell interplay in human chronic Chagas disease. Understanding the pathological immune mechanisms that sustain an inflammatory environment in human pathology is key to designing improved therapies.

The Microbiome of the Prostate Tumor Microenvironment.

BACKGROUND: The advent of molecular-based methods of identification and characterization of complex microbial populations has led to a new era of microbial discovery. A detailed and comprehensive analysis of the microbial ecosystem of the pathologic and healthy prostate tissues has not been yet reported. OBJECTIVES: To characterize the microbiome possibly associated to the pathologic prostate microenvironment. DESIGN, SETTING, AND PARTICIPANTS: The microbiome profile of tumor, peri-tumor, and nontumor tissues was assessed on 16 radical prostatectomy-specimens. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Microbiome analysis was assessed by massive ultradeep pyrosequencing. Bacteria load was expressed as a percentage of the total number of bacteria. The statistical significance of differences among specimen-groups was tested with Friedman's test (Dunn posthoc test) and Wilcoxon rank-sum test. RESULTS AND LIMITATIONS: Three phyla, six classes, nine orders, 14 families, and 11 genera were above the set threshold value of 1%, respectively. Significant differences in specific microbial populations among tumor/peri-tumor and nontumor prostate specimens were observed at certain taxonomic levels. Among genera, Propionibacterium spp. were the most abundant. Staphylococcus spp. were more represented in the tumor/peri-tumor tissues (p<0.05). The restricted number of specimens represents a potential limitation. CONCLUSIONS: The prostate contains a plethora of bacteria, which set themselves within the gland with a distribution dependent on the nature of the tissue, thus suggesting a possible pathophysiological correlation between the composition of the local microbial niche and the presence of the tumor itself. Future studies will help to clarify the role of these specific bacteria and their potential to be exploited as new biomarkers. PATIENT SUMMARY: The pathological prostate is populated by specific microbial populations, whose distribution varies according to the nature of the tissue. This finding opens interesting perspectives for the identification of novel therapeutic approaches and biomarkers.

IL-6 promotes M2 macrophage polarization by modulating purinergic signaling and regulates the lethal release of nitric oxide during Trypanosoma cruzi infection.

The production of nitric oxide (NO) is a key defense mechanism against intracellular pathogens but it must be tightly controlled in order to avoid excessive detrimental oxidative stress. In this study we described a novel mechanism through which interleukin (IL)-6 mediates the regulation of NO release induced in response to Trypanosoma cruzi infection. Using a murine model of Chagas disease, we found that, in contrast to C57BL/6 wild type (WT) mice, IL-6-deficient (IL6KO) mice exhibited a dramatic increase in plasma NO levels concomitant with a significantly higher amount of circulating IL-1ß and inflammatory monocytes. Studies on mouse macrophages and human monocytes, revealed that IL-6 decreased LPS-induced NO production but this effect was abrogated in the presence of anti-IL-1ß and in macrophages deficient in the NLRP3 inflammasome. In accordance, while infected WT myocardium exhibited an early shift from microbicidal/M1 to anti-inflammatory/M2 macrophage phenotype, IL6KO cardiac tissue never displayed a dominant M2 macrophage profile that correlated with decreased expression of ATP metabolic machinery and a lower cardiac parasite burden. The deleterious effects of high NO production-induced oxidative stress were evidenced by enhanced cardiac malondialdehyde levels, myocardial cell death and mortality. The survival rate was improved by the treatment of IL-6-deficient mice with a NO production-specific inhibitor. Our data revealed that IL-6 regulates the excessive release of NO through IL-1ß inhibition and determines the establishment of an M2 macrophage profile within infected heart tissue.

Natural Product Kuwanon-L Inhibits HIV-1 Replication through Multiple Target Binding.

In recent years many advances have been made in the fight against HIV-1 infection. However, the lack of a vaccine, together with the increasing resistance to the highly active anti-retroviral therapy (HAART), make HIV-1 infection still a serious global emergency. Thus, new compounds with original modes of action are continuously required, and natural products have ever been a very interesting class of pharmacologically active molecules. Some of them have been used since ancient times against viral infections. Here we present a work in which we suggest that kuwanon-L, a natural product active as an HIV-1 integrase (IN) inhibitor, might exert its overall antiviral activity through binding to multiple viral targets. Specific enzymatic tests, together with a time-of-addition (TOA) experiment, support our hypothesis of binding both to IN and to reverse transcriptase (RT). Overall, this compound can be considered an attractive lead for the development of new classes of antiviral agents able to overcome the problem of resistance, due to its ability to exert its action by binding simultaneously to multiple viral targets.

Adaptive immunity against gut microbiota enhances apoE-mediated immune regulation and reduces atherosclerosis and western-diet-related inflammation.

Common features of immune-metabolic and inflammatory diseases such as metabolic syndrome, diabetes, obesity and cardiovascular diseases are an altered gut microbiota composition and a systemic pro-inflammatory state. We demonstrate that active immunization against the outer membrane protein of bacteria present in the gut enhances local and systemic immune control via apoE-mediated immune-modulation. Reduction of western-diet-associated inflammation was obtained for more than eighteen weeks after immunization. Immunized mice had reduced serum cytokine levels, reduced insulin and fasting glucose concentrations; and gene expression in both liver and visceral adipose tissue confirmed a reduced inflammatory steady-state after immunization. Moreover, both gut and atherosclerotic plaques of immunized mice showed reduced inflammatory cells and an increased M2 macrophage fraction. These results suggest that adaptive responses directed against microbes present in our microbiota have systemic beneficial consequences and demonstrate the key role of apoE in this mechanism that could be exploited to treat immune-metabolic diseases.

IL-6 Improves the Nitric Oxide-Induced Cytotoxic CD8+ T Cell Dysfunction in Human Chagas Disease.

Reactive oxygen and nitrogen species are important microbicidal agents and are also involved in lymphocyte unresponsiveness during experimental infections. Many of the biological effects attributed to nitric oxide are mediated by peroxynitrites, which induce the nitration of immune cells, among others. Our group has demonstrated that nitric oxide is involved in the suppressive activity of myeloid-derived suppressor cells in Trypanosoma cruzi-infected mice, with a higher number of CD8+ T cells suffering surface-nitration compared to uninfected controls. Studying the functional and phenotypic features of peripheral CD8+ T cells from chagasic patients and human cells experimentally infected with T. cruzi, we found that different regulatory mechanisms impaired the effector functions of T cytotoxic population from seropositive patients. Peripheral leukocytes from chagasic patients showed increased nitric oxide production concomitant with increased tyrosine nitration of CD8+ T cells. Additionally, this cytotoxic population exhibited increased apoptotic rate, loss of the TCRζ-chain, and lower levels of CD107a, a marker of degranulation. Strikingly, IL-6 stimulation of in vitro-infected peripheral blood mononuclear cells obtained from healthy donors, blunted T. cruzi-induced nitration of CD3+CD8+ cells, and increased their survival. Furthermore, the treatment of these cultures with an IL-6 neutralizing antibody increased the percentage of T. cruzi-induced CD8+ T cell nitration and raised the release of nitric oxide. The results suggest that the under-responsiveness of cytotoxic T cell population observed in the setting of long-term constant activation of the immune system could be reverted by the pleiotropic actions of IL-6, since this cytokine improves its survival and effector functions.

Estado de vacunación y condición serológica del personal de salud de Córdoba, Argentina. Sector Público y Privado / Vaccination status and serologic condition in health personnel in Córdoba, Argentina

Introducción: el personal de salud (PS) tiene mayor riesgo de exposición/transmisión de enfermedades, siendo la vacunación un medio eficaz para reducirlo.Objetivos: evaluar estado de vacunación del lPS. Detectar factores de vacunación incompleta. Estado serológico (VHB, VHC, VIH).Material y métodos: estudio multicéntrico, observacional, descriptivo de corte transversal. Se encuestó 30 % del PS de 3 instituciones públi-cas y 2 privadas de Córdoba en marzo/abril de 2013. Recomendaciones Argentina: hepatitis B (HB), sarampión/rubéola (SR), difteria, tétanos, pertusis (dT/dTpa) e influenza. Análisis bivariado, prueba del Chi-cua-drado. Epidat 3.1.Resultados: total 676 encuestas. Esquema completo 20 % (estu-diantes 38 %, médicos 18 %, enfermeros 18 %, radiología 7 %, lim-pieza 5 %). Esquema vigente dT/dTpa: 78 %. Esquema HB: 64 %. In-fluenza 2012: 55 %. SR: 46 %. Motivos de vacunación incompleta: 28 % desconocía indicación, 23 % falta de interés, 2 % otros moti-vos, 2 % no acepta vacunación, 45 % no contestó. Conocen su con-dición serológica de VIH 63 %, VHB: 52 %, VHC: 44 %. Sector público (n = 546) vs privado (n = 130): esquema comple-to 21,24 % vs 14,61 % (p = 0,08). HB 66 % vs 54 % (p = 0,0099) dT/dTpa 81 % vs 65 % (p = 0,0001). SR 46 % vs 45 % (p = 0,87). Influenza 56 % vs 49 % (p = 0,12). Conclusión: el esquema de vacunación es incom-pleto en alto porcentaje de encuestados, principal-mente personal de limpieza; causa predominante: desconocimiento de indicación y desinterés. Ma-yor cobertura para dT/dTpa y HB. No hay diferen-cias entre sector público y privado respecto de es-quema completo. El sector público presenta mejor cobertura para HB y dT/dTpa, estadísticamente significativa. El test del VIH es el realizado con mayor frecuencia. Son necesarias campañas para concientizar y mejorar niveles de cobertura de vacunación

Introduction: Health personnel (HP) have higher risk of exposure/transmission of diseases, vaccination remains an effective means to reduce it.Objectives: Evaluate recommended vaccination in HP. Detect conditions of incomplete vaccination. Assess knowledge of their serological status: HBV, HCV, HIV.Material and Methods: Multicenter, observational, analitic, cross-sectional, conducted in 3 public hospitals and 2 private institutions Cba, Arg. We surveyed 10-30% of staff in each institution during March-April 2013. Recommendations Arg: Hepatitis B, Measles/Rubella (MR) Tetanus, diphtheria, pertussis (Td/Tdap) and Influenza. Bivariate analysis, using X2 test. Results: 676 surveys were completed. Complete vaccination: 20% (students 38%, physicians 18%, nurses 18%, lab 13%, radiology 7%, cleaners 5%). Current scheme for Td / Tdap: 78%. Hepatitis B:64%. Influenza vaccine: 55%, MR: 46%. Analysis by public (n=546) vs. private (n=130): Full scheme 21,24%vs14,61% (p=0.08). Hepatitis B 66%vs54%(p=0.0099). Td/Tdap 81%vs65% (p=0.0001). MR 46%vs45% (p=0.87). Influenza 56%vs49% (p=0.12). Causes of incomplete vaccination: 28% unknow this indication, 23% lack of interest. Know their serologic status: HIV 63%, HBV 52%, HCV 44%. Conclusion: High percentage of workers has incomplete vaccination.Lack of indication and lack of interest are the reasons most frequently detected. Better coverage for Td/Tdap and HB. Public or private sector does not predict better coverage but HB and Td/Tdapin public sector has a statistically significantly better coverag. Low percentage of respondents known their serological status. The HIV test is the most frequently performed. Designed campaigns are needed to increase vaccination coverageand to stimulating the knowledge of serological status of HP

Estado de vacunación en embarazadas y puérperas de un hospital público polivalente / Vaccination status in pregnant and puerperal women in a public general hospital

Introducción: conocer el estado de vacunación durante la gestación tiene por objeto la protección de madre e hijo. Objetivos: caracterizar epidemiológicamente a la población. Deter-minar el estado de vacunación de embarazadas/puérperas. Anali-zar la indicación de vacuna antigripal según época del año.Materiales y Métodos: estudio analítico. Encuesta en dos tiempos: previo a época invernal (grupo1, 13/2/13-13/3/13) y durante época invernal (grupo2, 5/7/13-15/8/13) en embarazadas/puérperas inter-nadas. Variables: demográficas, cobertura de doble adultos (dT)/triple bacteriana acelular (dTpa), antihepatitis B y antigripal.Resultados: total 437 pacientes, 47 embarazadas y 390 puérperas. Edad promedio 25 años (r = 14-48). Argentinas 89,5 %. Embarazos controlados 76 %. Escolaridad: secundario completo 21,5 %, in-completo 52,6 %. En unión estable 68 %. Embarazo no planifica-do 60,2 %. Vacuna dT: vigente: 81 %. dTpa: colocada 59,5 % (93 % durante el embarazo). Antihepatitis B: 50,3 % tenían la vacuna (91,8 % esque-ma completo). Cobertura antigripal total: 54,2 %. Grupo 1: 29 % vs Grupo 2, 86 % (p< 0,001). El 99 % se colocó la vacuna durante la gestación. Coberturas según embarazo controlado/no controla-do: dT 81,5 % / 73 % (p = 0,28), dTpa 61,8 % / 23 % (p = 0,0001), antihepatitis B 46,2 % / 46,1 % (p = 0,99) y antigripal 57 % / 11,5 % (p = 0,0000). Conclusión: predominaron mujeres jóvenes, con secundario incompleto, embarazo no planificado y deficiencias en su estado vacunal. La cobertura para dTpa y antigripal fue mayor en controladas con relación esta-dísticamente significativa. La vacuna antigripal tuvo mayor indica-ción en época invernal. Preocupa el desconocimiento sobre su estado de inmunización. Es importante la intervención del equipo de salud para motivar contro-les durante el embarazo, evaluar esquema de vacunación y educar sobre enfermedades inmunoprevenibles

Introduction: Knowing the status of vaccination during pregnancy aims at the protection of mother and child. Objectives: Epidemiologically characterize the population. To determine the vaccination status of pregnant/postpartum women. Analyze the indication of influenza vaccine according to the season. Materials and Methods: analytical study. Survey in two stages: pre-winter period (group 1, 13/2/13-13/3/13) and during winter time (Group 2, 5/7/13-15/8/13) in pregnant/postpartum women admitted. Variables: demographic, adults double coverage (dT)/acellular bacterial triple (DTaP), hepatitis B and influenza. Results: Total 437 patients, 47 pregnant and 390 postpartum women. Average age 25 years (r=14-48). Argentine 89.5%. Controlled pregnancies 76%. Schooling: Secondary full 21.5%, 52.6% incomplete. Cohabitants 68%. Unplanned pregnancy 60.2%. dT vaccine: current 81%. DTaP: placed 59.5% (93% during pregnancy). Hepatitis B: 50.3 % had the vaccine (91.8% complete scheme). Total influenza coverage: 54.2%. Group 1:29% vs Group 2, 86 % (p<0.001). 99% placed the vaccine during pregnancy. Hedges as controlled/uncontrolled pregnancy: dT 81.5 %/73 % (p=0.28), DTaP 61.8% / 23% (p =0.0001), hepatitis B 46.2 %/46.1% (p=0.99) and influenza 57%/11.5 % (p= 0.0000). Conclusion: Young women predominated, with incomplete secondary, unplanned pregnancy and deficiencies in their vaccination status. Coverage for DTaP and flu vaccine was higher in controlled pregnancies statistically significant relationship. The flu vaccine had greater indication winter. Ignorance about their immunization status is concerned. Intervention is important to the health team to motivate controls during pregnancy, evaluate vaccination and education on preventable diseases

Prognostic value of circulating cytokines on overall survival and disease-free survival in cancer patients.

Through their tumor-promoting and/or tumor-suppressive properties, cytokines can influence progression of cancer. We systematically reviewed the current literature on the prognostic value of the circulating cytokines IL-1α/ß, IL-6, IL-8, IL-10, IL-12, TNF-α, TGF-ß and IFN-γ to predict overall and disease-free survival in any type of cancer patients. PubMed was systematically searched and based on eligibility assessment using our five criteria of the Reporting Recommendations for Tumor Marker Prognostic Studies (REMARK) checklist, six unique studies were reviewed. Elevated IL-6 and IL-10 levels seem independently associated with worse prognosis in terms of overall and disease-free survival. The prognostic value of IL-1α/ß, IL-8, IL-12, TNF-α, TGF-ß and IFN-γ could not be demonstrated. The small number of selected studies underlines the need for large well-designed prospective studies, using the REMARK checklist as a guideline, to determine which cytokines have prognostic value on survival in cancer patients.

Compromiso respiratorio en embarazadas: puérperas hospitalizadas en el contexto de la Pandemia 2009 de Influenza A (H1N1)pdm09 / Respiratory compromise in hospitalised pregnant women: postpartum during Pandemic Influenza A (H1N1) pdm09, 2009

Introducción: el diagnóstico de influenza en gestantes está asociado a un alto riesgo de morbi-mortalidad. Objetivos: describir características clínico-epidemiológicas de embarazadas/puérperas cursando enfermedad tipo influenza (ETI) y el compromiso respiratorio durante la pandemia de influenza A (H1N1) pdm09. Comparar severidad según edad gestacional. Material y métodos: estudio retrospectivo, descriptivo mediante revisión de historias clínicas de embarazadas/puérperas internadas por ETI en dos maternidades, período: 27/06/09-14/08/09. Se tabularon en dos grupos: G1, n= 35 (1º y 2º trimestre); G2, n= 49 (3º trimestre/puerperio). Resultados: se incluyeron 84 pacientes, edad promedio 25 años (R: 14-42). 81% sin comorbilidades. 20% ingresaron a Unidad de Terapia Intensiva (UTI). El 54% consultó tardíamente. Presentaron neumonía el 50%. Recibió tratamiento antiviral el 89%. El 23% terminó su embarazo durante la internación por indicación obstétrica. El 94% del total presentó evolución favorable. Comparando G1 vs G2: necesidad de UTI (5/35 vs 12/49, p=0.25), roncus/sibilancias (8/35 vs. 18/49, p= 0,17), saturación de oxígeno menor de 96 % (6/35 vs 15/49 p=0,15), número de muertes (1/35 vs. 4/49, p= 0,30). Internación prolongada (3/35 vs 14/49, p= 0,02). La mortalidad general fue 6%. Conclusiones: la mayoría de las pacientes no presentaba comorbilidades y se encontraba cursando el tercer trimestre de embarazo o puerperio al momento de la internación. Hubo mayor frecuencia de compromiso respiratorio severo, internación en UTI y mortalidad en las pacientes en este grupo, sin diferencia significativa respecto al primer y segundo trimestre de embarazo. La mortalidad fue comparable a la reportada en la bibliografía.

Introduction: diagnosis of influenza in pregnant women is associated with a high risk of morbidity and mortality. Objectives: To describe clinical and epidemiological characteristics and respiratory compromise of hospitalized pregnant/postpartum women suffering from influenza like illness, assisted during the pandemic of influenza A (H1N1)pdm09 and compare serverity of respiratory compromise according to gestational age. Material and methods: retrospective, descriptive study through a review of medical charts of pregnant/postpartum women asisted in two maternity hospitals, period: 27/June to 14/August 2009. The data was tabulated into two groups: G1, n=35 (1st and 2nd Trimester), and G2, n=49, 3rd Trimester/puerperium. Results: 84 patients was included. Average age 25 years (R: 15-42). 81% without comorbidities. 20% was admitted on Intensive Care Unit (ICU). Pneumonia was diagnosed in 50%. 89% received antiviral treatment. 23% ended their pregnancy during the hospitalization for obstetric indication. 94% of the population presents favorable clinical evolution. Comparing G1 vs. G2: need for ICU admission (5/35 vs 12/49, p=0.25), presence of rhonchi/wheezing (8/35 vs 18/49, p=0.17), oxygen saturation < 96% (6/35 vs 15/49, p=0.15), mortality (1/35 vs 4/49, p=0.30), prolonged hospitalization (3/35 vs 14/49, p=0.02). Overall mortality was 6%. Conclusions: Most patientes had not comorbidities and was enrolled in the third trimester of pregnancy or puerperium at the time of hospitalization. There was a higher frequency of severe respiratory compromise, hospitalization in ICU and mortality in patients in this group, without significant difference compared to the first and second trimester. Mortality was comparable to that reported in the literature.

Overexpression of 17ß-hydroxysteroid dehydrogenase type 1 increases the exposure of endometrial cancer to 17ß-estradiol.

CONTEXT: The local interconversions between estrone (low activity) and 17ß-estradiol (potent compound) by 17ß-hydroxysteroid dehydrogenases (17ß-HSDs) can lead to high 17ß-estradiol generation in endometrial cancer (EC). OBJECTIVE: Examine the balance between the 17ß-HSDs reducing estrone to 17ß-estradiol (types 1, 5, 12, and 7) and those oxidizing 17ß-estradiol to estrone (2, 4, and 8), in EC. PATIENTS AND METHODS: Reducing and oxidizing 17ß-HSD activities (HPLC) and mRNA level (RT-PCR) were assessed in normal post-menopausal (n = 16), peritumoral endometrium (normal tissue beside cancer, n = 13), and 58 EC (29 grade 1, 18 grade 2, 11 grade 3). RESULTS: Grade 1 EC displayed a shifted estrone reduction/17ß-estradiol oxidation balance in favor of 17ß-estradiol compared with controls. This was more pronounced among estrogen receptor-α (ER-α)-positive biopsies. Type 1 17ß-HSD mRNA (HSD17B1 gene expression, real time PCR) and protein levels (immunohistochemistry) were higher in ER-α-positive grade 1 EC than controls. The mRNA coding for types 4, 5, 7, 8, and 12 17ß-HSD did not vary, whereas that coding for type 2 17ß-HSD was increased in high-grade lesions compared with controls. Three-dimensional ex vivo EC explant cultures demonstrated that 17ß-HSD type 1 generated 17ß-estradiol from estrone and increased tumor cell proliferation. Additional in vitro studies using EC cells confirmed that in the presence of 17ß-HSD type 1, estrone induced estrogen signaling activation similarly to 17ß-estradiol. Therefore, estrone was reduced to 17ß-estradiol. CONCLUSIONS: Type 1 17ß-HSD increases 17ß-estradiol exposure in grade 1 EC, thus supporting tumor growth. This enzyme represents a potential therapeutic target.