Prevalence and risk factors of Strongyloides stercoralis in haemodialysis in Cochabamba, Bolivia: a cross-sectional study.

BACKGROUND: Strongyloidiasis is an infectious disease that can be fatal in immunocompromised patients. Patients with end-stage renal failure who are on dialysis have a considerably weakened immune system, and organ transplantation is a major risk factor for severe strongyloidiasis. Knowledge of the local epidemiology in tropical and subtropical areas is an essential prerequisite for designing an appropriate strategy to prevent this potentially lethal complication. In this study, we aimed to estimate the prevalence and associated risk factors of S. stercoralis infection in patients on dialysis in Cochabamba, Bolivia. METHODS: A cross-sectional study was carried out among patients undergoing haemodialysis in Cochabamba (elevation 2,500 m, temperate climate), collecting information on socio-demographic, lifestyle, and clinical variables, and using one coproparasitological technique (the modified Baermann technique) and one serological (ELISA) test for S.stercoralis diagnosis. RESULTS: In total, 149 patients participated in the study (mean age = 51.4 years, 48.3% male). End-stage renal disease was predominantly (59%) of hypertensive and/or diabetic origin. The positive serological prevalence was 18.8% (95% CI: 13.3%-25.9%). Based on the sensitivity and specificity of the ELISA test, the estimate of the actual prevalence was 15.1% (95% CI: 9.4%-20.7%). Stool samples of 105 patients (70.5%) showed a coproparasitological prevalence of 1.9% (95% CI: 0.52%-6.68%). No potential risk factors were significantly associated with S. stercoralis infection. CONCLUSIONS: We found a high seroprevalence of S. stercoralis in Bolivian patients undergoing haemodialysis in Cochabamba. We recommend presumptive antiparasitic treatment at regular intervals to avoid the potentially fatal complications of severe strongyloidiasis.

Clonotypic IgH Response against Systemic Viral infection in Pronephros and Spleen of a Teleost Fish.

Upon infection, B lymphocytes develop clonal responses. In teleost fish, which lack lymph nodes, the kinetics and location of B cell responses remain poorly characterized. Fish pronephros is the site of B cell differentiation and the main niche for persistence of plasma cells. In this study, we undertook the analysis of the rainbow trout IgHµ repertoire in this critical tissue for humoral adaptive immunity after primary immunization and boost with a rhabdovirus, the viral hemorrhagic septicemia virus (VHSV). We used a barcoded 5' RACE-cDNA sequencing approach to characterize modifications of the IgHµ repertoire, including VH usage in expressed V(D)J rearrangements, clonal diversity, and clonotype sharing between individual fish and treatments. In the pronephros, our approach quantified the clonotype frequency across the whole IgH repertoire (i.e., with all VH), measuring the frequency of Ag-responding clonotypes. Viral infection led to extensive modifications of the pronephros B cell repertoire, implicating several VH subgroups after primary infection. In contrast, only modest changes in repertoire persisted 5 mo later, including VHSV-specific public expansions. The IgM public response implicating IgHV1-18 and JH5, previously described in spleen, was confirmed in pronephros in all infected fish, strongly correlated to the response. However, the distribution of top clonotypes showed that pronephros and spleen B cells constitute distinct compartments with different IgH repertoires. Unexpectedly, after boost, the frequency of anti-VHSV clonotypes decreased both in pronephros and spleen, raising questions about B cell circulation. A better monitoring of B cell response kinetics in lymphoid tissues will be an essential step to understand B memory and plasmocyte formation mechanisms in fish.

Effect of CaS Nanostructures in the Proliferation of Human Breast Cancer and Benign Cells In Vitro.

We report on the effect of naked CaS nanostructures on the proliferation of carcinoma cancer cells and normal fibroblasts in vitro. The CaS nanostructures were prepared via the microwave-mediated decomposition of dimethyl sulfoxide (DMSO) in the presence of calcium acetate Ca ( CH 3 CO 2 ) 2 . Light scattering measurements revealed that dispersions contain CaS nanostructures in the size range of a few Å to about 1 nanometer, and are formed when DMSO is decomposed in the presence of Ca ( CH 3 CO 2 ) 2 . Theoretical calculations at the DFT/B3LYP/DGDZVP level of theory on ( C a S ) n clusters ( n = 1 , 2 , 3 , and 4) are consistent with clusters in this size range. The absorption spectra of the CaS nanostructures are dominated by strong bands in the UV, as well as weaker absorption bands in the visible. We found that a single dose of CaS nanoclusters smaller than 0.8 nm in diameter does not affect the survival and growth rate of normal fibroblasts and inhibits the proliferation rate of carcinoma cells in vitro. Larger CaS nanostructures, approximately (1.1 ± 0.2) nm in diameter, have a similar effect on carcinoma cell proliferation and survival rate. The CaS nanoclusters have little effect on the normal fibroblast cell cycle. Human carcinoma cells treated with CaS nanocluster dispersion exhibited a decreased ability to properly enter the cell cycle, marked by a decrease in cell concentration in the G0/G1 phase in the first 24 h and an increase in cells held in the SubG1 and G0/G1 phases up to 72 h post-treatment. Apoptosis and necrotic channels were found to play significant roles in the death of human carcinoma exposed to the CaS nanoclusters. In contrast, any effect on normal fibroblasts appeared to be short-lived and non-detrimental. The interaction of CaS with several functional groups was further investigated using theoretical calculations. CaS is predicted to interact with thiol ( R-SH ), hydroxide ( R - OH ), amino ( R - NH 2 ), carboxylic acid ( R - COOH ), ammonium ( R-NH 3 + ), and carboxylate ( R-COO - ) functional groups. None of these interactions are predicted to result in the dissociation of CaS. Thermodynamic considerations, on the other hand, are consistent with the dissociation of CaS into Ca 2 + ions and H 2 S in acidic media, both of which are known to cause apoptosis or cell death. Passive uptake and extracellular pH values of carcinoma cells are proposed to result in the observed selectivity of CaS to inhibit cancer cell proliferation with no significant effect on normal fibroblast cells. The results encourage further research with other cell lines in vitro as well as in vivo to translate this nanotechnology into clinical use.

HIV and Chagas Disease: An Evaluation of the Use of Real-Time Quantitative Polymerase Chain Reaction to Measure Levels of Trypanosoma cruzi Parasitemia in HIV Patients in Cochabamba, Bolivia.

This cross-sectional study evaluated epidemiologic characteristics of persons living with HIV (PWH) coinfected with Trypanosoma cruzi in Cochabamba, Bolivia, and estimated T. cruzi parasitemia by real-time quantitative polymerase chain reaction (qPCR) in patients with and without evidence of reactivation by direct microscopy. Thirty-two of the 116 HIV patients evaluated had positive serology for T. cruzi indicative of chronic Chagas disease (27.6%). Sixteen of the 32 (50%) patients with positive serology were positive by quantitative polymerase chain reaction (qPCR), and four of the 32 (12.5%) were positive by direct microscopy. The median parasite load by qPCR in those with CD4+ < 200 was 168 parasites/mL (73-9951) compared with 28.5 parasites/mL (15-1,528) in those with CD4+ ≥ 200 (P = 0.89). There was a significant inverse relationship between the degree of parasitemia estimated by qPCR from blood clot and CD4+ count on the logarithmic scale (rsBC= -0.70, P = 0.007). The correlation between T. cruzi estimated by qPCR+ blood clot and HIV viral load was statistically significant with rsBC = 0.61, P = 0.047. Given the significant mortality of PWH and Chagas reactivation and that 57% of our patients with CD4+ counts < 200 cells/mm3 showed evidence of reactivation, we propose that screening for chronic Chagas disease be considered in PWH in regions endemic for Chagas disease and in the immigrant populations in nonendemic regions. Additionally, our study showed that PWH with advancing immunosuppression have higher levels of estimated parasitemia measured by qPCR and suggests a role for active surveillance for Chagas reactivation with consideration of treatment with antitrypanosomal therapy until immune reconstitution can be achieved.

Cutting Edge: Neutralizing Public Antibody Responses Are an Ancient Form of Defense Conserved in Fish and Mammals.

The repertoire of Abs is generated by genomic rearrangements during B cell differentiation. Although V(D)J rearrangements lead to repertoires mostly different between individuals, recent studies have shown that they contain a substantial fraction of overrepresented and shared "public" clones. We previously reported a strong public IgHµ clonotypic response against the rhabdovirus viral hemorrhagic septicemia virus in a teleost fish. In this study, we identified an IgL chain associated with this public response that allowed us to characterize its functionality. We show that this public Ab response has a potent neutralizing capacity that is typically associated with host protection during rhabdovirus infections. We also demonstrate that the public response is not restricted to a particular trout isogenic line but expressed in multiple genetic backgrounds and may be used as a marker of successful vaccination. Our work reveals that public B cell responses producing generic Abs constitute a mechanism of protection against infection conserved across vertebrates.

Effect of jasmonic acid on major terpenes and density of glandular trichomes in Lippia graveolens kunth (Verbenaceae).

The effect of exogenous application of jasmonic acid (JA) on the concentration of main terpenes and density of glandular trichomes was investigated in the Mexican oregano, propagated from seeds from 3 localities. JA 1 mM was applied locally and to the whole plant. JA locally applied increased the number of trichomes, with a mean of 20 trichomes more with respect to the controls in plants from Tecomavaca and Zapotitlán Salinas, and significantly increased the thymol concentration by 185% systemically and 255% locally, compared to the control. JA applied to the whole plant decreased the number of trichomes and increased the concentration of caryophyllene from 0.79 to 1.7 mg g-1, and α-caryophyllene from 0.3 to 0.8 mg g-1 in plants from San Rafael with reference to water control. The results suggest a plasticity of morphologic and phytochemical responses, and a potential use of JA to improve phenolic monoterpenes and sesquiterpenes production.

Standardized IMGT® Nomenclature of Salmonidae IGH Genes, the Paradigm of Atlantic Salmon and Rainbow Trout: From Genomics to Repertoires.

In teleost fish as in mammals, humoral adaptive immunity is based on B lymphocytes expressing highly diverse immunoglobulins (IG). During B cell differentiation, IG loci are subjected to genomic rearrangements of V, D, and J genes, producing a unique antigen receptor expressed on the surface of each lymphocyte. During the course of an immune response to infections or immunizations, B cell clones specific of epitopes from the immunogen are expanded and activated, leading to production of specific antibodies. Among teleost fish, salmonids comprise key species for aquaculture. Rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) are especially important from a commercial point of view and have emerged as critical models for fish immunology. The growing interest to capture accurate and comprehensive antibody responses against common pathogens and vaccines has resulted in recent efforts to sequence the IG repertoire in these species. In this context, a unified and standardized nomenclature of salmonid IG heavy chain (IGH) genes is urgently required, to improve accuracy of annotation of adaptive immune receptor repertoire dataset generated by high-throughput sequencing (AIRRseq) and facilitate comparisons between studies and species. Interestingly, the assembly of salmonids IGH genomic sequences is challenging due to the presence of two large size duplicated IGH loci and high numbers of IG genes and pseudogenes. We used data available for Atlantic salmon to establish an IMGT standardized nomenclature of IGH genes in this species and then applied the IMGT rules to the rainbow trout IGH loci to set up a nomenclature, which takes into account the specificities of Salmonid loci. This unique, consistent nomenclature for Salmonid IGH genes was then used to construct IMGT sequence reference directories allowing accurate annotation of AIRRseq data. The complex issues raised by the genetic diversity of salmon and trout strains are discussed in the context of IG repertoire annotation.

Spleen and head kidney differential gene expression patterns in trout infected with Lactococcus garvieae correlate with spleen granulomas.

Lactococcus garvieae is a significant pathogen in aquaculture with a potential zoonotic risk. To begin to characterize the late immune response of trout to lactococcosis, we selected infected individuals showing clinical signs of lactococcosis. At the time lactococcosis clinical signs appeared, infection by L. garvieae induced a robust inflammatory response in the spleen of rainbow trout, which correlated with abundant granulomatous lesions. The response in kidney goes in parallel with that of spleen, and most of the gene regulations are similar in both organs. A correlation existed between the early inflammatory granulomas in spleen (containing macrophages with internalized L. garvieae) and up-regulated gene sets, which defined the presence of macrophages and neutrophils. This is the first analysis of the immune transcriptome of rainbow trout following L. garvieae infection during the initiation of adaptive immune mechanisms and shows a transcriptome induction of antibody response by both IgM (+) and IgT (+) spleen B cells to respond to systemic infection. These results increase our understanding of lactococcosis and pave the way for future research to improve control measures of lactococcosis on fish farms.

Micropapillary Cervical Adenocarcinoma: A Clinicopathologic Study of 44 Cases.

Micropapillary adenocarcinoma has been reported as an aggressive variant of adenocarcinoma in several organs, where it is associated with poor clinical outcome. This study reports the clinicopathologic features and outcomes of cervical adenocarcinomas with a micropapillary component (micropapillary cervical adenocarcinomas); this represents the largest reported study of these neoplasms. The study comprised 44 cervical adenocarcinomas of usual (human papillomavirus-related)-type (84%), mucinous, not otherwise specified (4.5%), gastric-type (4.5%), endometrioid (4.5%), and adenosquamous carcinoma (2%). The micropapillary component comprised >50% of the neoplasm in 34 cases (77%) (group 1), and 10% to 50% in 10 cases (23%) (group 2). Lymph node metastasis was present in 41 of 44 (93%) cases and typically the nodal tumor retained a prominent micropapillary morphology. Follow-up ranged from 7 to 123 months (mean, 65.9 mo). Seventeen of 44 (38.6%) patients had no evidence of disease on follow-up, 6/44 (13.6%) were alive with disease, and 21/44 (47.7%) died of disease. There were no survival differences between group 1 and group 2. On univariate analysis, lymph node metastasis (P=0.0015), lymphovascular space invasion (P=0.002), parametrial involvement (P=0.03), and depth of stromal invasion (P=0.045) were related to tumor recurrence. On multivariate analysis, lymph node metastasis (P=0.001), and extent of lymphovascular space invasion (P=0.027) were significant independent predictors of tumor recurrence. Our study shows that a micropapillary component in cervical adenocarcinoma may be associated with aggressive behavior and that a micropapillary architecture may occur within a variety of types of cervical adenocarcinoma.

Epidemiology of Strongyloides stercoralis infection in Bolivian patients at high risk of complications.

BACKGROUND: Strongyloidiasis can be fatal in immunocompromised patients, but few epidemiological studies investigated the burden of this neglected tropical disease among these populations, particularly in low- and middle-income countries such as Bolivia. This study aimed to fill in this gap by estimating prevalence rate and risk factors associated with strongyloidiasis among patients at high risk of complications. METHODS: A cross-sectional study was carried out in Santa Cruz (elevation 400 meters, tropical climate) and Cochabamba (elevation 2,500 meters, temperate climate), among patients with cancer, HIV infection and rheumatic or hematologic disease, using four coproparasitological techniques and one serological (ELISA) test. RESULTS: In total, 1,151 patients participated in this study, including individuals who were HIV-positive (30%) or with rheumatic (29%), oncologic (32%) or hematologic (9%) diseases. The serological and coproparasitological prevalence was 23.0% (95% confidence interval [CI], 20.7-25.5; n = 265/1151) and 7.6% (95% CI, 6.2-9.3; n = 88/1151), respectively, with an estimated actual prevalence of 20.2% (95% CI, 17.9-22.5). Positive serology and positive coproparasitology were associated with younger age and lower education levels. There was no significant difference in prevalence between Cochabamba and Santa Cruz as defined by coproparasitology (6.4% vs. 8.9%; p = 0.11) or serology (24.0% vs. 22.0%; p = 0.4). Among 64 patients in Cochabamba who had never travelled to the tropical lowlands, 5 (7.8%) had a positive coproparasitology. CONCLUSIONS: Strongyloidiasis is widely prevalent in Bolivia among vulnerable patients at increased risk of life-threatening complications. Transmission of the parasite occurs both in tropical lowlands and temperate elevation (≥ 2,500 m). Control strategies to prevent transmission and complications of this serious parasitic disease should be urgently reinforced.

Caracterización clínico-epidemiológica de pacientes con leptospirosis en el municipio Cumanayagua. Provincia Cienfuegos. 2007-2017 / Clinical-epidemilogical characterization of patients with leptopirosis in the Cumanayagua Municipality. Cienfuegos Province. 2007-2017

Fundamento: la leptospirosis continua siendo la zoonosis más frecuente a nivel mundial. Objetivo: caracterizar clínica y epidemiológicamente pacientes con leptospirosis. Métodos: investigación observacional descriptiva que incluyó los 38 casos notificados con leptospirosis en el municipio Cumanayagua de la provincia Cienfuegos, en el período 2007-2017. Las variables que se utilizaron fueron: año y mes del diagnóstico, edad, sexo, procedencia, antecedentes de vacunación, posible fuente de infección, signos, síntomas y complicaciones. Los datos fueron recogidos de las series cronológicas y de las historias epidemiológicas dispuestas en la Unidad Municipal de Higiene y Epidemiología del municipio. El procesamiento de los datos se realizó en el programa Excel y se utilizaron porcientos y tasas de incidencia. Resultados: se notificaron el período 2072 pacientes sospechosos de la enfermedad y solo hubo 38 confirmados. La tasa más elevada estuvo en el año 2013 con 22,5 por 100 000 habitantes. El grupo de edad de 25 a 59 años fue el más afectado con un 42,1%. Solo el 15 % tenía antecedentes de haber sido vacunados con la vacuna antileptospírica. La mayor fuente de infección fue el estar vinculado a labores agrícolas con un 47,4 % seguido del contacto con animales para un 28,9 %. Dentro de los síntomas más frecuentes estuvieron la fiebre, cefalea, mialgias y artralgias. Conclusiones: se evidenció subregistro en el diagnóstico serológico de los casos, riesgo ocupacional con vínculo hacia las labores agrícolas y predominio de la forma clínica anictérica.

Foundation: leptospirosis still is the most prequent zoonosis worldwide. Objective: to characterize clinically and epidemiologically patients with leptospirosis. Methods: Observational descriptive method which included 38 cases diagnosed leptospirosis in the Cumanayagua Municipality, Cienfuegos Province in the period 2207- 2017. The variables used were year and month of the diagnosis, age sex, precedence, vaccination antecedents, possible source of infection, signs, symptoms and complications. Data was collected from chronological series and epidemiological histories at the Municipal Center of Hygiene and Epidemiology. Data processing was done with Excel and percentages and incidence rates were used. Results: during the period 2072 patients were reported as suspected to have the disease and only 38 were confirmed. The highest rate was in 2013 with 42.1%. Only 15% had antecedents of being vaccinated with the anti- leptospirosis vaccine. The infection source of higher incidence was to be an agricultural worker with 47.4%, followed by contact with animals 28.9%. Among the most frequent symptoms were fever, chephalalgias, myalgias and arthralgias. Conclusion: a sub-registration in the serological diagnosis of the cases., occupational risks with contact to agricultural activities and predominance of clinical anicteric form.

Guía para la enseñanza del apoyo vital socorrista. Consenso para el proyecto de formación e investigación en apoyo vital. Cienfuegos, 2018 / Guideline for teaching life support. Agreement for the project of training and research in life support. Cienfuegos, 2018

Fundamento: la parada cardiaca primaria es un problema de salud mundial y una de sus soluciones es disponer, tanto la población como los profesionales de la salud, de conocimientos en reanimación cardiopulmocerebral. El proyecto para la formación e investigación en apoyo vital en emergencias y desastres puede ser un marco académico para lograrlo. Objetivo: actualizar, para el proyecto, las guías y estrategia docente para la enseñanza del socorrismo. Métodos: taller nacional realizado en julio del 2018 en Cienfuegos, con 13 expertos y cuatro informantes clave. Se utilizaron la técnica de tormenta de ideas y de grupo nominal, cuestionario semiestructurado y revisión documental previa. Resultados: se proponen para la enseñanza del socorrismo tanto en el pregrado como el posgrado: los objetivos, aspectos didácticos, estrategia docente, contenidos, alcance, habilidades a desarrollar, futuras investigaciones y áreas para la colaboración interinstitucional. Se definió la estructura de un curso prototipo para la enseñanza del tema y las principales consideraciones para su ejecución. Conclusiones: las propuestas académicas para el curso "Apoyo vital socorrista" permiten el abordaje de la enseñanza de los primeros auxilios tanto a la población como a grupos especializados de socorristas.

Foundation: Primary cardiac failure is a worldwide Health problem and one of its solutions is to count on health professionals and the population in general with knowledge in cardio-pulmonary-cerebral resuscitation. The project for the training and research in life support in emergencies and disasters may be an academic framework to achieve it. Objective: to update, for the project, the teaching guides and strategy for first aids. Methods: national workshop developed in July 2018 in Cienfuegos, with 13 experts and four key informers. The techniques of brain storming, and nominal group, semi-structured questionnaire and documentary review were used. Results: it is proposed for teaching first aids to undergraduate students and postgradutes as well: the objectives, specific aspects, teaching strategy, contents, scope, skills to develop, future research, and areas for inter-institutional collaboration. Conclusion: academic proposals for the course "Life Support" allows approaching treaching first aids to the population and specialized groups as well.

Guía para la enseñanza del apoyo vital avanzado ante múltiples víctimas. Consenso para el proyecto de formación e investigación en apoyo vital. Cienfuegos, 2018 / Guideline for teaching advanced life support in the presence of mass casualty. Consensus for the project of training and research in life support. Cienfuegos, 2018

Fundamento: en situaciones de desastres, cuando el número de enfermos o lesionados es alto y los recursos limitados, la atención médica precisa de cambios de estructura y procesos si se quiere salvar un número elevado de personas. El personal de salud debe estar entrenado para enfrentar esta situación cada vez más frecuente. El "Proyecto para la formación e investigación en apoyo vital en emergencias y desastres" puede ser un marco académico para lograrlo. Objetivo: actualizar, para el proyecto, las guías y estrategia docente para la enseñanza del apoyo vital ante víctimas múltiples. Métodos: taller nacional realizado el 10-11 de julio del 2018 en Cienfuegos, con 13 expertos y cuatro informantes clave. Se utilizaron la técnica de tormenta de ideas y de grupo nominal, cuestionario semiestructurado y revisión documental previa. Resultados: se proponen para la enseñanza del apoyo vital en situaciones de desastres, tanto en el pregrado como el posgrado: los objetivos, aspectos didácticos, estrategia docente, contenidos, alcance, habilidades a desarrollar, futuras investigaciones y áreas para la colaboración interinstitucional. Se definió la estructura de un curso prototipo para la enseñanza del tema y las principales consideraciones para su ejecución. Conclusiones: las propuestas académicas para el curso "Apoyo vital avanzado ante víctimas múltiples" permiten la preparación de los profesionales de la salud para brindar asistencia médica en situaciones de desastres, con escasos recursos y en ambientes complejos.

Foundation: in disaster situations, when the number of patients or injured is high and the resources are limited, medical care requires changes of structures and processes if it is aimed to save most of the persons involved. Health personnel should be trained to face this situation becoming more frequent each time. Objective: to update, for the project, the guidelines and strategies for teaching life support in the presence of mass casualty. Methods: national workshop developed in July 10th and 11th in Cienfuegos, with 13 experts and four key informers. The techniques of brain storming and nominal group, semi-structured and previous documentary review. Results: objectives, specific aspects, teaching strategy, contents, scope of the abilities, skills to develop, future research, and areas for inter-institutional collaboration were proposed for teaching life support in situations of disaster. Conclusion: academic proposals for the course "Life support in the presence of mass casualty¨ allow preparing health professionals to offer medical assistance in situations of disasters with limited resources in complex environments.

Caracterización clínico-epidemiológica de pacientes con tuberculosis en el municipio Cumanayagua. Provincia Cienfuegos. 2007-2017 / Clinical-epidemiological characterization of tuberculosis in the Cumanayagua Municiality. Cienfuegos Province. 2007-2017

Fundamento: en Cuba constituye hoy un reto eliminar la tuberculosis. Objetivo: caracterizar clínica y epidemiológicamente pacientes con tuberculosis. Métodos: investigación observacional descriptiva que incluyó los 24 casos notificados con tuberculosis en el municipio Cumanayagua de la provincia Cienfuegos, en el período 2007-2017. Las variables que se analizaron fueron: año del diagnóstico, clasificación según tipo de tuberculosis (pulmonar y extrapulmonar), edad, sexo, lugar del diagnóstico, ocupación, consejo popular, área de salud, factores de riesgo asociados, signos y síntomas. Los datos fueron recogidos de las series cronológicas y de las historias epidemiológicas dispuestas en la Unidad Municipal de Higiene y Epidemiología del municipio. El procesamiento de los datos se realizó en el programa Excel y se utilizaron porcientos y tasas de incidencia. Resultados: la mayor tasa de la enfermedad se presentó en el año 2013. El mayor número de casos se reportó en los pacientes de 55 a 59 años con nueve. Predominaron los pacientes con baciloscopia positiva con un 66,7 %. El 62,5 % fue diagnosticado en el segundo nivel de atención. Predominó el sexo masculino, jubilados y obreros agrícolas. Los factores de riesgo más frecuentes fueron el tabaquismo y alcoholismo. Los síntomas más frecuentes fueron la tos, pérdida de peso, astenia y fiebre. Conclusiones: la tuberculosis en el municipio Cumanayagua se presentó de acuerdo a las características clínico epidemiológicas descritas en la literatura revisada; hubo dificultades en la oportunidad del diagnóstico en el primer nivel de atención.

Introduction: It is a challange to eliminate tuberculosis in Cuba. Objective: to characterize clinically and epidemiologically pacients with tuberculosis. Methods: observational descriptive research which included 24 cases reported with tuberculosis in Cumanayagua from 2007 to 2017. Tha variables analized were: year of diagnosis, classification according to the type of tuberculosis (pulmonary or extra-pulmonary), Age, sex, place of diagnosis, occupation, community, health area, associated risk factors, signs and symptoms. Data was collected from the chronological series and epidemiological histories existing at the Municipal Units of Hygiene and Epidemilogy in the municipality. Data processing was done by excell and percentages and incidence rate were used. Results: the highest rate of the disease presented in 2013. The highest number of cases was reported from 55 to 59 years old with 9 cases. Patients with positive basiloscopy with a 66,7 %. A 62,5 % was diagnosed in the second level of care. The male sex, retired and agriculture workers predominated. The most frequent risk factors were tabaquism and alcoholism. The most frequent symptoms were cough, loss of weight, asthenia and fiver. Conclusion: tuberculosis at the Municipality of Cumanayagua presented according to the clinical and epidemiological characteristics described in the literature reviewed; there were difficulties in the opportunity of diagnosis at the first care level.

B Cell-Activating Factor Regulates Different Aspects of B Cell Functionality and Is Produced by a Subset of Splenic B Cells in Teleost Fish.

In mammals, B cell functionality is greatly influenced by cytokines released by innate cells, such as macrophages or dendritic cells, upon the early recognition of common pathogen patterns through invariant receptors. B cell-activating factor (BAFF) is one of these innate B cell-helper signals and plays a key role in the survival and differentiation of B cells. Although, evolutionarily, teleost fish constitute the first animal group in which adaptive immunity based on Ig receptors is present, fish still rely greatly on innate responses. In this context, we hypothesized that BAFF would play a key role in the control of B cell responses in fish. Supporting this, our results show that teleost BAFF recapitulates mammalian BAFF stimulating actions on B cells, upregulating the expression of membrane MHC II, improving the survival of fish naïve B cells and antibody-secreting cells, and increasing the secretion of IgM. Surprisingly, we also demonstrate that BAFF is not only produced in fish by myeloid cells but is also produced by a subset of splenic B cells. Thus, if this B cell-produced BAFF proves to be actively regulating this same B cell subset, our findings point to an ancient mechanism to control B cell differentiation and survival in lower vertebrates, which has been silenced in mammals in physiological conditions, but reemerges under pathological conditions, such as B cell lymphomas and autoimmune diseases.

The BAFF / APRIL axis plays an important role in virus-induced peritoneal responses in rainbow trout.

IgM+ B cells have been recently demonstrated to be key regulators of peritoneal inflammation in teleost, as a large number of them occupy the peritoneal cavity after 48 h of antigenic stimulation. Despite this, the number of studies addressing the mechanism through which this cell population expands and differentiates in response to stimuli has been scarcely addressed. Because the BAFF/APRIL axis is known to play a major role in B cell survival and differentiation in mammals, we hypothesized that it could be affected in the peritoneal cavity in response to an inflammatory stimulus. To verify this hypothesis, we studied how BAFF, APRIL and the fish-specific related cytokine BALM as well as their putative receptors are regulated in rainbow trout after intraperitoneal (i.p.) injection of viral hemorrhagic septicemia virus (VHSV). When the transcriptional analysis was performed in total cells from the peritoneum, we observed that VHSV provoked an up-regulation of both BAFF and BAFF receptor (BAFF-R) mRNA levels. However, when we examined how isolated peritoneal IgM+ B cells were transcriptionally affected by VHSV i.p. injection, we found that APRIL, BALM and the transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) were also up-regulated in response to the virus. IgM- cells, on the other hand, only up-regulated BALM transcription in response to VHSV. Finally, to gain further insight on the role that these cytokines play in the peritoneum, we have studied their effect on the survival of peritoneal IgM+ B cells. This work demonstrates a key role for the BAFF/APRIL axis in the peritoneal inflammatory response and contributes to further understanding how IgM+ B cells are regulated at this specific peripheral site.

Describing the diversity of Ag specific receptors in vertebrates: Contribution of repertoire deep sequencing.

During the last decades, gene and cDNA cloning identified TCR and Ig genes across vertebrates; genome sequencing of TCR and Ig loci in many species revealed the different organizations selected during evolution under the pressure of generating diverse repertoires of Ag receptors. By detecting clonotypes over a wide range of frequency, deep sequencing of Ig and TCR transcripts provides a new way to compare the structure of expressed repertoires in species of various sizes, at different stages of development, with different physiologies, and displaying multiple adaptations to the environment. In this review, we provide a short overview of the technologies currently used to produce global description of immune repertoires, describe how they have already been used in comparative immunology, and we discuss the future potential of such approaches. The development of these methodologies in new species holds promise for new discoveries concerning particular adaptations. As an example, understanding the development of adaptive immunity across metamorphosis in frogs has been made possible by such approaches. Repertoire sequencing is now widely used, not only in basic research but also in the context of immunotherapy and vaccination. Analysis of fish responses to pathogens and vaccines has already benefited from these methods. Finally, we also discuss potential advances based on repertoire sequencing of multigene families of immune sensors and effectors in invertebrates.

Expansion and differentiation of IgM+ B cells in the rainbow trout peritoneal cavity in response to different antigens.

To date, intraperitoneal (i.p.) injection seems to be the most effective vaccination route in aquaculture, as many i.p. administered fish vaccines are capable of conferring strong and long-lasting immune responses. Despite this, how peritoneal leukocytes are regulated upon antigen encounter has only been scarcely studied in fish. Although, in the past, myeloid cells were thought to be the main responders to peritoneal inflammation, a recent study revealed that IgM+ B cells are one of the main cell types in the teleost peritoneal cavity in response to pathogenic bacteria. Thus, in the current work, we have focused on establishing how IgM+ B cells are recruited into the peritoneum in rainbow trout (Oncorhynchus mykiss) comparing different antigens: Escherichia coli as a bacterial model, E. coli-derived lipopolysaccharide (LPS) or viral hemorrhagic septicemia virus (VHSV). In addition to studying their capacity to dominate the peritoneal cavity, we have established how these IgM+ B cells are regulated in response to the different antigens, determining their levels of IgM secretion, surface MHC II expression, cell size and phagocytic abilities. Our results reveal that IgM+ B cells are one of the main cell types amplified in the peritoneum in response to either bacterial or viral antigens and that these immunogenic stimulations provoke a differentiation of some of these cells towards plasmablasts/plasma cells whereas others seem to be implicated in antigen presentation. These findings contribute to a better understanding of the immune processes that regulate peritoneal inflammation in teleost fish.

Distinct Differentiation Programs Triggered by IL-6 and LPS in Teleost IgM(+) B Cells in The Absence of Germinal Centers.

Although originally identified as a B cell differentiation factor, it is now known that mammalian interleukin-6 (IL-6) only regulates B cells committed to plasma cells in response to T-dependent (TD) antigens within germinal centers (GCs). Even though adaptive immunity is present in teleost fish, these species lack lymph nodes and GCs. Thus, the aim of the present study was to establish the role of trout IL-6 on B cells, comparing its effects to those induced by bacterial lipopolysaccharide (LPS). We demonstrate that the effects of teleost IL-6 on naïve spleen B cells include proliferation, activation of NF-κB, increased IgM secretion, up-regulation of Blimp1 transcription and decreased MHC-II surface expression that point to trout IL-6 as a differentiation factor for IgM antibody-secreting cells (ASCs). However, LPS induced the secretion of IgM without up-regulating Blimp1, driving the cells towards an intermediate activation state in which antigen presenting mechanisms are elicited together with antibody secretion and expression of pro-inflammatory genes. Our results reveal that, in trout, IL-6 is a differentiation factor for B cells, stimulating IgM responses in the absence of follicular structures, and suggest that it was after follicular structures appeared that this cytokine evolved to modulate TD responses within the GC.

Cross Talk Between Growth and Immunity: Coupling of the IGF Axis to Conserved Cytokine Pathways in Rainbow Trout.

Although disease and infection is associated with attenuated growth, the molecular pathways involved are poorly characterized. We postulated that the IGF axis, a central governor of vertebrate growth, is repressed during infection to promote resource reallocation towards immunity. This hypothesis was tested in rainbow trout (Oncorhynchus mykiss) challenged by Aeromonas salmonicida (AS), a Gram-negative bacterial pathogen, or viral hemorrhagic septicemia virus (VHSv) at hatch, first feeding, and 3 weeks after first feeding. Quantitative transcriptional profiling was performed for genes encoding both IGF hormones, 19 salmonid IGF binding proteins (IGFBPs) and a panel of marker genes for growth and immune status. There were major differences in the developmental response of the IGF axis to AS and VHSv, with the VHSv challenge causing strong down-regulation of many genes. Despite this, IGFBP-1A1 and IGFBP-6A2 subtypes, each negative regulators of IGF signaling, were highly induced by AS and VHSv in striking correlation with host defense genes regulated by cytokine pathways. Follow-up experiments demonstrated a highly significant coregulation of IGFBP-1A1 and IGFBP-6A2 with proinflammatory cytokine genes in primary immune tissues (spleen and head kidney) when trout were challenged by a different Gram-negative bacterium, Yersinia ruckeri. Based on our findings, we propose a model where certain IGFBP subtypes are directly regulated by cytokine signaling pathways, allowing immediate modulation of growth and/or immune system phenotypes according to the level of activation of immunity. Our findings provide new and comprehensive insights into cross talk between conserved pathways regulating teleost growth, development, and immunity.