Etiología y caracterización epidemiológica del síndrome febril no palúdico en tres municipios del Urabá antioqueño, Colombia

Introduction: Urabá, a region on the northern coast of Colombia, is endemic to several acute febrile illnesses of infectious origin; however, only patients with malaria may have access to quick and effective diagnosis. For this reason, many non-malarial febrile patients go without a clear etiologic diagnosis.Aim: To establish the etiology and clinical signs of acute febrile non-malaria syndromes and explore some of the likely risk factors in patients originating in the municipalities of Necocli, Turbo and Apartado who exhibit these symptoms.Materials and methods: We obtained acute and convalescent sera from 220 non-malarial febrile patients from the rural and urban zones of Necocli, Turbo and Apartado during 2007 and 2008. Serologic tests for dengue (IgM by ELISA), leptospirosis (IgM and IgG by IFA), rickettsiosis (IgG by IFI), hanta and arenavirus (IgG by ELISA) were performed.Results: We found that the frequency of infection for dengue, leptospirosis, rickettsiosis and arenavirus, was 37.3%; 14.1%; 2.7% and 0.5%, respectively. There were 12 co-infection cases of leptospirosis-dengue and one of leptospirosis-rickettsiosis-dengue. Male gender and relative humidity were considered risk factors for dengue, and the beginning of clinical signs in February of 2008 was associated with the infection of dengue and leptospirosis.Conclusion: This study confirms previous records that underline the importance of Rickettsia spp,dengue virus and Leptospira spp as causal agents of febrile syndrome in this region of Colombia. doi: http://dx.doi.org/10.7705/biomedica.v33i0.734Introducción. La región de Urabá es endémica para varias enfermedades febriles agudas de origen infeccioso. Sin embargo, solo los pacientes con malaria pueden acceder a un diagnóstico oportuno y rápido, motivo por el cual muchos síndromes febriles no palúdicos quedan sin diagnóstico etiológico claro.Objetivo. Establecer la etiología, describir las manifestaciones clínicas y explorar algunos posibles factores de riesgo relacionados con los síndromes febriles agudos no palúdicos en pacientes procedentes de los municipios de Necoclí, Turbo y Apartadó.Materiales y métodos. Se tomaron muestras de suero en fase aguda y de convalecencia de 220 pacientes febriles negativos para malaria, provenientes de zonas rurales y urbanas de Necoclí, Turbo y Apartadó en los años 2007 y 2008. Se practicaron pruebas para diagnóstico de dengue (detecciónde anticuerpos IgM por ELISA), leptospirosis (detección de anticuerpos IgM e IgG por IFI), rickettsiosis (detección de anticuerpos IgG por IFI), hantavirus y arenavirus (detección de anticuerpos IgG porELISA).Resultados. Se encontraron frecuencias de dengue, leptospirosis, rickettsiosis y arenavirus de 37,3 %,14,1 %, 2,7 % y 0,5 %, respectivamente. Se presentaron 12 casos de coinfección de leptospirosis-denguey uno de leptospirosis-rickettsiosis-dengue. El sexo masculino y la humedad relativa media,fueron factores de riesgo para dengue. El inicio de signos clínicos en febrero de 2008, se asoció tanto con la infección por dengue como por leptospirosis.Conclusión. Se reafirma la importancia del virus del dengue, Rickettsia spp. y Leptospira spp., como agentes causantes del síndrome febril en la región del Urabá. doi: http://dx.doi.org/10.7705/biomedica.v33i0.734

[autosomal Chronic Granulomatous Disease: Case Report And Mutation Analysis Of Two Brazilian Siblings].

To report the case of two siblings with chronic granulomatous disease. Chronic granulomatous disease is a primary immunodeficiency disorder characterized by abnormal microbicidal activity. Mutations in the p47-phox gene (NCF-1) are present in about 30% of the patients with chronic granulomatous disease; this group presents a better prognosis and later onset of recurrent infections as compared with the X-linked variant, present in about 56% of patients. Case 1 is a female presenting repeat infections since age 10, starting with impetigo followed by severe pneumonia six months later. The severity of the lung infection associated with liver abscess and the patient's resistance to treatment prompted laboratory investigation for immunodeficiency. The results of the nitroblue tetrazolium and superoxide release tests were consistent with a diagnosis of chronic granulomatous disease. The parents and siblings were assessed, revealing the presence of granulomatous disease in a brother (Case 2). He also presented repeat infections with impetigo at age 10, followed by pneumonia six months later, however in a non severe form. Single-strand conformational polymorphism analysis detected abnormal electrophoretic mobility of exon 2 of the NCF-1 gene. Sequence DNA analysis revealed a dinucleotide GT deletion in exon 2. It is important to evaluate the relatives of chronic granulomatous disease patients, even in the absence of typical clinical signs. Defining the mutation and its correlation with phenotype is important to provide appropriate genetic counseling and clinical prognosis.80425-

Isolation of human bone marrow mesenchymal stem cells and evaluation of their osteogenic potential

Las células madre mesenquimatosas de médula ósea humana (abreviadas hBMSCs) constituyen una fuente de células auto-renovables con alto potencial de diferenciación, comúnmente aisladas a partir de los aspirados medulares en huesos largos. Su diferenciación hacia el linaje osteogénico, por ejemplo, ha sido ampliamente utilizada para la evaluación biológica de biomateriales o matrices con aplicaciones en la ingeniería de tejidos óseos. El objetivo de este trabajo consistió en aislar hBMSCs a partir de la cabeza femoral de pacientes sometidos a artroplastia total de cadera, así como evaluar su potencial osteogénico. Brevemente, se extrajo el hueso esponjoso y se disgregó mecánicamente; las células desprendidas se cultivaron y las células no adherentes se eliminaron luego de 4 días. El potencial osteogénico se evaluó en la quinta generación de cultivo, mediante ensayos de diferenciación a 14 y 20 días donde se compararon cultivos con y sin suplementos osteogénicos. La evaluación se realizó mediante tinción con Alizarina Roja y la cuantificación de los niveles de expresión génica de los marcadores osteogénicos colágeno tipo I, osteonectinca y sialoprotiena ósea mediante RT-PCR en tiempo real. Las hBMSCs obtenidas presentaron un fenotipo no-diferenciado estable, así como la capacidad de mineralizar la matriz extracelular y expresar un fenotipo similar al osteoblasto durante la inducción osteogénica. Los tres marcadores evaluados se sobre-expresaron en los cultivos en condiciones osteogénicas, y se encontró que cambios hasta de 2X en sus niveles de expresión son relevantes para el desarrollo del proceso de diferenciación. El modelo de hBMSCS presentado podría ser utilizado para la evaluación in vitro de la osteoinductividad de diferentes biomateriales, moléculas bioactivas o matrices para ingeniería de tejidos.Human bone marrow mesenchymal stem cells (hBMSCs) comprise a cell population capable of self-renewal and multilineage differentiation commonly isolated from bone marrow aspirates of large bones. Their osteogenic potential has been extensively exploited for the biological evaluation of scaffolds or biomaterials with applications in bone tissue engineering. This work aimed to isolate hBMSCs from femoral heads of patients undergoing total hip arthroplasty and to evaluate their osteogenic potential. Briefly, the trabecular bone was extracted and mechanically disaggregated; the released cells were cultured and non-adherent cells were removed after 4 days. The osteogenic potential was evaluated at the fifth passage after 14 and 20 days of induction, comparing cultures with and without osteogenic supplements, via Alizarin red staining and the quantification of the gene expression levels of the osteogenic markers collagen type I, osteonectin and bone sialoprotein through real-time RT-PCR. The obtained hBMSCs presented a stable undifferentiated phenotype after prolonged cell culture, matrix mineralization capabilities and expression of osteoblast phenotype upon osteogenic induction. The three markers were up-regulated in cultures under osteogenic conditions and 2 fold differences in their expression levels were found to be significant for the onset of the differentiation process. The obtained hBMSCs may have applications on the in vitro evaluation of the osteoinductivity of different biomaterials, bioactive molecules or tissue engineering scaffolds

Leptospirosis humana en Colombia: la experiencia del Instituto Colombiano de Medicina Tropical - CES

Leptospirosis is an emerging infectious disease that is transmitted from animals to humans in both rural and urban settings. This report presents the experience of Colombian Institute of Tropical Medicine-CES, during the period of 2000-2005; we determined the frequency of antibodies for 14 serovar of Leptospira spp mainly with the serovars Grippotyphosa and Copenhageni. In the Embera-Katio's indigenous community from Cordoba, a transversal study based on enzyme-linked immunosorbert assay (ELISA), the seroprevalence was 18,1 % In the Colombian region of Uraba the seroprevalence was 12,5 %. In conclusion there is a high prevalence of Leptospira spp antibodies and it is thus necessary to establish control measures in order to decrease the risk of environmental exposure to leptospirosis in the region.La leptospirosis ha venido aumentando su frecuencia en todo el mundo, tanto en humanos como en animales, así como en los ambientes rurales y los urbanos. En este trabajo se presenta la experiencia entre 2000 y 2005 del Instituto Colombiano de Medicina Tropical - CES. Durante el período de estudio se encontraron anticuerpos para 14 serovoariedades de Leptospira spp, principalmente para las serovariedades Grippotyphosa y Copenhageni. En un estudio realizado en una comunidad indígena de Córdoba, utilizando el método de ELISA, la seroprevalencia fue de 18,1 %. En otro  trabajo en población urbana de la región de Urabá, se encontró una prevalencia de 12,5 %. Estos resultados confirman la circulación de Leptospira spp en nuestro medio, y dada su importancia clínica y epidemiológica, es necesario lograr la integración entre los organismos de salud animal y humana, para controlar la leptospirosis y estudiar otros aspectos de la enfermedad que aun son desconocidos

Leptospirosis: Diagnóstico serológico

Leptospirosis is a globally important zoonotic disease that affects humans on all continents, in both urban and rural contexts, and in temperate and tropical climates. Leptospirosis is a disease of the environment; transmission depends on interactions between humans and mammalian reservoir hosts. A variety of infectious diseases that present as undifferentiated febrile syndromes, such as malaria, dengue and influenza can mimic Leptospirosis. Differential diagnosis and clinical suspicion of the similar presentations of these diseases are necessary as well as identify predominant risk factors and to improve he leptospirosis laboratory~based surveillance system for the early detection of patients with febrile syndromes.La leptospirosis es una enfermedad zoonótica ampliamente distribuida, que afecta a la pobla¬ción humana en todos los continentes, en ambientes tanto urbanos como rurales y en climas templados y tropicales. La leptospirosis es una enfermedad que procede del ambiente y la transmisión depende de la interacción entre humanos y animales reservorios. Una variedad de enfer¬medades infecciosas que se presentan como síndromes febriles indiferenciados, tales como malaria, influenza y dengue pueden simular leptospirosis, por lo que un diagnóstico diferencial entre estas etiologías se debe considerar. Es necesario determinar también, posibles factores de riesgo de esta enfermedad y reforzar la vigilancia centinela de síndromes febriles desarrollando la capacidad diagnóstica en los laboratorios de Salud Pública, incluyendo las pruebas serológicas para Leptospira como parte de los estudios rutinarios en pacientes febriles

Frecuencia de anticuerpos para 14 serovariedades de Leptospira spp detectados por la prueba de microaglutinación en una serie de casos humanos de Antioquia, Colombia

Leptospirosis is a zoonotic disesase of great incidence intropical regions due to environmental, climatic and socialfactors that favor its transmission. The microagglutinationtest (MAT) is the reference diagnostic for the WorldOrganization of the Health, test is also used for epidemicstudies to be a test serovar specific, it can specify theserovars that are circulating in human and animalpopulations, indispensable fact when is necessary toestablish the bases to propose measures of control of theillness. To determine the frequency of antibodies for 14serovars of Leptospira spp in 49 individuals that consultedto the Colombian Institute of Tropical Medicine-CES,during the period of 2000-2006, to confirm theleptospirosis diagnosis. The 49 individuals had an age 34.2 year-old average, 53,8 % was men. 70 % was residentsin the Medellin city and other Antioquia municipalities,of which 30 % came from Apartado in the regionNorth West of Antioquia state. The MAT revealed that67,34 % of the serums agglutinated with the serovarGrippotyphosa; 63,2 % for the serovar Copenhageni;40,81 % belonged to the serovar Icterohaemorragiae; 6,12 %for the serovar Tarassovi and 2,04 % presented agglutinationsagainst L. biflexa. This study allowed thepresence of Leptospira and the serovars that are infectingthe population from Antioquia state and it demonstratesthat there are several circulating serovars.  La leptospirosis es una zoonosis de gran incidencia en regiones tropicales debido a factores ambientales, climáticos y sociales que favorecen su transmisión. La microaglutinación (MAT) es la prueba diagnóstica de referencia para la Organización Mundial de la Salud, la prueba se emplea además para estudios epidemiológicos por ser una prueba serovar específica, es decir, que puede precisar las serovariedades que están circulando en poblaciones humanas y animales, dato indispensable cuando se plantea establecer las bases para proponer medidas de control de la enfermedad. Se propuso determinar la frecuencia de anticuerpos para 14 serovariedades de Leptospira spp en 49 individuos que consultaron al Instituto Colombiano de Medicina Tropical – CES, durante el periodo de 2000-2006, para confirmar el diagnóstico de leptospirosis. Los 49 individuos tuvieron una edad promedio de 34,2 años, el 53,8 % fueron hombres. El 70 % eran residentes en la ciudad de Medellín y otros municipios antioqueños, de los cuales el 30 % procedían del municipio de Apartadó en la región noroccidental del departamento. La prueba de MAT reveló que 67,34 % de los sueros aglutinaron con el serovar Grippotyphosa; 63,2 % para el serovar Copenhageni; 40,81 % pertenecían al serovar Icterohaemorragiae; 6,12 % para el serovar Tarassovi y 2,04 % presentaron aglutinaciones contra L. biflexa. Este estudio permitió establecer la presencia de Leptospira y las serovariedades que están infectando a la población humana residente en el departamento de Antioquia y demuestra que hay múltiples serovariedades circulantes en esta población

Canine Leptospirosis in a Northwestern Region of Colombia: Serological, Molecular and Epidemiological Factors

Canine leptospirosis is a zoonosis of epidemiological importance. Dogs are recognized as primary reservoirs of Leptospira interrogans serogroup Canicola and a source of infection to the environment through urine. This study aimed to determine the presence of antibodies against Leptospira in canines from 49 municipalities in the Department of Antioquia, Colombia. We performed a cross-sectional study of dogs included in a neutering control program. We collected 1335 sera samples, assayed by a microagglutination test (MAT), and performed PCR detection in 21 urine samples. We also surveyed 903 dog owners. We found a seroreactivity of 11.2% (150/1335) in Antioquia with titers ≥1:50. Municipalities with the highest number of cases were Belmira (46.1%), Turbo (34.5%), and Concepción (31.0%). L. santarosai was identified by phylogenetic analysis in one urine sample from the municipality of Granada. The most important factor associated with a positive result was the lack of vaccination against leptospirosis (PR 3.3, p ≤ 0.014). Environmental factors such as water presence and bare soil around the household were also associated with Leptospira seroreactivity in the Department of Antioquia. We reviewed a national epidemiological surveillance database for human cases in those municipalities. We found a correlation between the high number of cases in canines and humans, especially in the Uraba. Serological and molecular results showed the circulation of Leptospira. Future public health efforts in the municipalities with the highest numbers of seroreactivity should be directed towards vaccination to prevent animal disease and decrease the probability of transmission of Leptospira. Dogs actively participate in the Leptospira cycle in Antioquia and encourage the implementation of vaccination protocols and coverage

Chromoblastomycosis Murine Model and In vitro Test to Evaluate the Sensitivity of Fonsecaea pedrosoi to Ketoconazole, Itraconazole and Saperconazole

An experimental model of murine chromoblastomycosis and in vitro tests with Fonsecaea pedrosoi were used to test the sensitivity of this fungus to three different antimycotics. The experimental model was standardized in BALB/c mice inoculated intraperitoneally with a 10^6 CFU/ml suspension of a F. pedrosoi isolate. Clinical infection was evident after 5 days of inoculation. Three groups of 27 mice each were used in the experiment. One group was treated with ketoconazole (KTZ), another with itraconazole (ITZ) and the other with saperconazole (SPZ). Antimycotic therapy was continued for 21 days. The control group consisted of 40 mice which were inoculated, but not treated. Infection was documented by macroscopic and microscopic examination of affected tissue in addition to culture of tissue macerates. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) for the F. pedrosoi strain used were done. The in vitro results showed that SPZ was the most active with MIC 0.01 mg/ml and MFC 0.1 mg/ml, followed by ITZ. SPZ was also the most effective in vivo since 63% of the treated animals (p=0.01) showed a curative effect after the observation period. We concluded that SPZ had the best in vitro and in vivo activity against F. pedrosoi

Inmunología de la leptospirosis

Leptospirosis is a re-emergent disease of worldwide distribution caused by pathogenic spirochetes of the Leptospira genus that affect humans, domestic and wild animals. The clinical manifestations of the disease are diverse and are the result of the interaction of the immune response of the host and the virulence conditions of the pathogenic species. Although many aspects of immunity in infection with Leptospira spp are unknown, it is recognized that susceptible hosts show differences in their immune response, such as activation / evasion of the complement system, activation of cellular subpopulations, production of cytokines, development of antibodies. Study of the immunological profile in patients with leptospirosis has been documented and contributes in the identification of bio-markers associated with severity. This review presents updated events related to the immune response from the entry of the bacteria in the initial phase of the infection to its multiplication and generation of human disease.Leptospirosis es una enfermedad re-emergente de distribución mundial ocasionada por espiroquetas patogénicas del género Leptospira que afectan humanos, animales domésticos o silvestres. Las manifestaciones clínicas de la enfermedad son diversas y son el resultado de la interacción de la respuesta inmune del hospedador y las condiciones de virulencia propias de las especies patógenas. Aunque se desconocen muchos aspectos de la inmunidad en la infección por Leptospira spp, es reconocido que los hospederos susceptibles presentan diferencias en su respuesta inmune, como la activación/evasión del sistema del complemento, la activación de subpoblaciones celulares, la producción de citoquina y el desarrollo de anticuerpos. El estudio del perfil inmunológico en pacientes con leptospirosis ha sido documentado y contribuye en la identificación de biomarcadores asociados con severidad. Esta revisión presenta algunos de los eventos relacionados con la respuesta inmune desde el ingreso de la bacteria en la fase inicial de la infección hasta su multiplicación y generación de enfermedad en el human