Seroprevalence of bovine viral diarrhea virus (BVDV) in cattle from Sotaquirá, Colombia

Worldwide distributed Bovine Viral Diarrhea Virus (BVDV) represents a high risk of infection in most bovine farms, in which it is associated with gastrointestinal, respiratory, and reproductive diseases. The purpose of this research was to establish the seroprevalence and the main risk factors associated with the presentation of BVDV in the municipality of Sotaquirá, Colombia. Samples were taken from 1000 cattle of Holstein, Ayrshire, Jersey, Normande Gyr and Holstein x Gyr. Epidemiological surveys were implemented, reproductive and management variables were taken into consideration. Indirect ELISA was performed to detect specific antibodies against BVDV using the commercial kit SERELISA® BVD p80 Ab Mono Blocking. The overall seroprevalence of antibodies against BVDV was 42.5% (425/1000), where the Gyr breed (59.1% apparent prevalence (AP); 60.3% real prevalence (PR)) and the age group > 4 years (53.0% PA; 54.4% PR) presented the highest seroprevalences. A significant statistical association was found for the breed, age, management practices evaluated and the presentation of PI3 (p ≤ 0.05). Age group > 4 years, Normande breed, presentation of PI3 and grazing lease were established as risk factors associated with BVDV in the herds. These infections are mainly associated with dairy cattle and herds with many animals, so it is important to consider vaccination plans as a preventive system and follow up on the most common diseases

Nuclear Translocation of b-Catenin during Mesenchymal Stem Cells Differentiation into Hepatocytes Is Associated with a Tumoral Phenotype

Wnt/b-catenin pathway controls biochemical processes related to cell differentiation. In committed cells the alteration of this pathway has been associated with tumors as hepatocellular carcinoma or hepatoblastoma. The present study evaluated the role of Wnt/b-catenin activation during human mesenchymal stem cells differentiation into hepatocytes. The differentiation to hepatocytes was achieved by the addition of two different conditioned media. In one of them, b-catenin nuclear translocation, up-regulation of genes related to the Wnt/b-catenin pathway, such as Lrp5 and Fzd3, as well as the oncogenes c-myc and p53 were observed. While in the other protocol there was a Wnt/b-catenin inactivation. Hepatocytes with nuclear translocation of b-catenin also had abnormal cellular proliferation, and expressed membrane proteins involved in hepatocellular carcinoma, metastatic behavior and cancer stem cells. Further, these cells had also increased auto-renewal capability as shown in spheroids formation assay. Comparison of both differentiation protocols by 2D-DIGE proteomic analysis revealed differential expression of 11 proteins with altered expression in hepatocellular carcinoma. Cathepsin B and D, adenine phosphoribosyltransferase, triosephosphate isomerase, inorganic pyrophosphatase, peptidyl-prolyl cis-trans isomerase A or lactate dehydrogenase b-chain were up-regulated only with the protocol associated with Wnt signaling activation while other proteins involved in tumor suppression, such as transgelin or tropomyosin b-chain were downregulated in this protocol. In conclusion, our results suggest that activation of the Wnt/b-catenin pathway during human mesenchymal stem cells differentiation into hepatocytes is associated with a tumoral phenotyp

Detección de Fasciola hepatica por medio de ELISA indirecto en ovinos y caprinos de Boavita, Colombia

The aim of this study was to determine the seroprevalence of Fasciola hepatica by indirect ELISA in sheep and goats from Boavita (Boyaca, Colombia). A descriptive cross-sectional study with simple random sampling was carried out. A total of 297 blood samples from sheep and 337 from goats of different breeds and age groups were collected. The sera were analyzed by the indirect ELISA technique with the commercial ELISA kit BIO K 211 - Monoscreen AbELISA F. hepatica. Additionally, an epidemiological survey was carried out. The seroprevalence of F. hepatica was 67.34% (200/297) in sheep and 59.94% (202/337) in goats. Likewise, seroprevalence was higher in males (sheep: 77.78%, 21/27; goats: 63.89%, 23/36), in sheep older than 3 years (83.33%, 50/60) and in goats less than 1 year of age. (69.77%, 60/86), as well as in Creole breed (sheep: 69.84%; goats: 61.79%). Sheep over 3 years of age were determined as a risk factor, while in goats the Creole breed variables and extensive grazing were identified as risk factors.El objetivo del estudio fue determinar la seroprevalencia de Fasciola hepatica por medio de ELISA indirecto en ovinos y caprinos de Boavita (Boyacá, Colombia). Se realizó un estudio descriptivo de corte transversal con muestreo aleatorio simple. Se colectaron 297 muestras de sangre de ovinos y 337 de caprinos de diferentes grupos raciales y etarios. Los sueros fueron analizados mediante la técnica ELISA indirecta con el kit comercial ELISA BIO K 211 - Monoscreen AbELISA F. hepatica. Adicionalmente se realizó una encuesta epidemiológica. La seroprevalencia de F. hepatica fue de 67.34% (200/297) en ovinos y de 59.94% (202/337) en caprinos. Asimismo, la seroprevalencia fue mayor en los machos (ovinos: 77.78%. 21/27; caprinos: 63.89%, 23/36), en ovinos mayores de 3 años (83.33%, 50/60) y en caprinos menores de 1 año (69.77%, 60/86), así como en los de raza criolla (ovinos: 69.84%; caprinos: 61.79%). Los ovinos mayores de 3 años se determinaron como factor de riesgo, en tanto que en los caprinos las variables raza criolla y el pastoreo extensivo se identificaron como factores de riesgo. El estudio demuestra una alta seropositividad de la enfermedad en la zona

Diagnóstico coproparasitológico de fascioliasis en ovinos y caprinos de Boavita, Boyacá (Colombia)

The aim of this study was to establish the prevalence of Fasciola hepatica through coprological analysis and to identify risk factors associated with the presentation of the parasite in sheep and goats in the municipality of Boavita, Boyacá. The study was observational, descriptive, cross-sectional with simple random sampling. Faecal samples were taken from 297 sheep and 337 goats to identify parasite eggs. The general prevalence was 8.0% (51/634), being 9.1% for sheep and 7.1% for goats. The prevalence in sheep was higher in males (14.8%) than in females (8.5%), likewise, sheep less than one year old (9.3%) and Criolla (11.1%) presented the highest prevalence. In goats, the prevalence was higher in females (7.3%) than in males (5.6%), and those less than one year old (10.5%) and the Alpine breed (8%) presented the greater prevalence. No significant statistical association was found between females and males. The Creole breed was established as a risk factor for sheep.El objetivo del estudio fue establecer la prevalencia de Fasciola hepatica mediante análisis coprológico e identificar factores de riesgo asociados a la presentación del parásito en ovinos y caprinos del municipio de Boavita, Boyacá. El estudio fue observacional, descriptivo de corte transversal con muestreo aleatorio simple. Se tomaron muestras de materia fecal a 297 ovinos y 337 caprinos para identificar los huevos del parásito. La prevalencia general fue de 8.0% (51/634), siendo de 9.1% para ovejas y de 7.1% para cabras. La prevalencia en ovinos fue mayor en machos (14.8%) que en hembras (8.5%); asimismo, los ovinos menores a un año (9.3%) y de Criolla (11.1%) presentaron las prevalencias más altas. En las cabras, la prevalencia fue mayor en hembras (7.3%) que en machos (5.6%), y los menores a un año (10.5%) y de la raza Alpina (8%) presentaron las prevalencias más altas. No se encontró asociación estadística significativa entre hembras y machos. La raza Criolla se estableció como factor de riesgo para los ovinos

Short-course combination treatment for experimental chronic Chagas disease

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, affects millions of people in the Americas and across the world, leading to considerable morbidity and mortality. Current treatment options, benznidazole (BNZ) and nifurtimox, offer limited efficacy and often lead to adverse side effects because of long treatment durations. Better treatment options are therefore urgently required. Here, we describe a pyrrolopyrimidine series, identified through phenotypic screening, that offers an opportunity to improve on current treatments. In vitro cell-based washout assays demonstrate that compounds in the series are incapable of killing all parasites; however, combining these pyrrolopyrimidines with a subefficacious dose of BNZ can clear all parasites in vitro after 5 days. These findings were replicated in a clinically predictive in vivo model of chronic Chagas disease, where 5 days of treatment with the combination was sufficient to prevent parasite relapse. Comprehensive mechanism of action studies, supported by ligand-structure modeling, show that compounds from this pyrrolopyrimidine series inhibit the Qi active site of T. cruzi cytochrome b, part of the cytochrome bc1 complex of the electron transport chain. Knowledge of the molecular target enabled a cascade of assays to be assembled to evaluate selectivity over the human cytochrome b homolog. As a result, a highly selective and efficacious lead compound was identified. The combination of our lead compound with BNZ rapidly clears T. cruzi parasites, both in vitro and in vivo, and shows great potential to overcome key issues associated with currently available treatments.</p

Short-course combination treatment for experimental chronic Chagas disease

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, affects millions of people in the Americas and across the world, leading to considerable morbidity and mortality. Current treatment options, benznidazole (BNZ) and nifurtimox, offer limited efficacy and often lead to adverse side effects because of long treatment durations. Better treatment options are therefore urgently required. Here, we describe a pyrrolopyrimidine series, identified through phenotypic screening, that offers an opportunity to improve on current treatments. In vitro cell-based washout assays demonstrate that compounds in the series are incapable of killing all parasites; however, combining these pyrrolopyrimidines with a subefficacious dose of BNZ can clear all parasites in vitro after 5 days. These findings were replicated in a clinically predictive in vivo model of chronic Chagas disease, where 5 days of treatment with the combination was sufficient to prevent parasite relapse. Comprehensive mechanism of action studies, supported by ligand-structure modeling, show that compounds from this pyrrolopyrimidine series inhibit the Qi active site of T. cruzi cytochrome b, part of the cytochrome bc1 complex of the electron transport chain. Knowledge of the molecular target enabled a cascade of assays to be assembled to evaluate selectivity over the human cytochrome b homolog. As a result, a highly selective and efficacious lead compound was identified. The combination of our lead compound with BNZ rapidly clears T. cruzi parasites, both in vitro and in vivo, and shows great potential to overcome key issues associated with currently available treatments.</p

Nuclear Translocation of β-Catenin during Mesenchymal Stem Cells Differentiation into Hepatocytes Is Associated with a Tumoral Phenotype

Wnt/β-catenin pathway controls biochemical processes related to cell differentiation. In committed cells the alteration of this pathway has been associated with tumors as hepatocellular carcinoma or hepatoblastoma. The present study evaluated the role of Wnt/β-catenin activation during human mesenchymal stem cells differentiation into hepatocytes. The differentiation to hepatocytes was achieved by the addition of two different conditioned media. In one of them, β-catenin nuclear translocation, up-regulation of genes related to the Wnt/β-catenin pathway, such as Lrp5 and Fzd3, as well as the oncogenes c-myc and p53 were observed. While in the other protocol there was a Wnt/β-catenin inactivation. Hepatocytes with nuclear translocation of β-catenin also had abnormal cellular proliferation, and expressed membrane proteins involved in hepatocellular carcinoma, metastatic behavior and cancer stem cells. Further, these cells had also increased auto-renewal capability as shown in spheroids formation assay. Comparison of both differentiation protocols by 2D-DIGE proteomic analysis revealed differential expression of 11 proteins with altered expression in hepatocellular carcinoma. Cathepsin B and D, adenine phosphoribosyltransferase, triosephosphate isomerase, inorganic pyrophosphatase, peptidyl-prolyl cis-trans isomerase A or lactate dehydrogenase β-chain were up-regulated only with the protocol associated with Wnt signaling activation while other proteins involved in tumor suppression, such as transgelin or tropomyosin β-chain were down-regulated in this protocol. In conclusion, our results suggest that activation of the Wnt/β-catenin pathway during human mesenchymal stem cells differentiation into hepatocytes is associated with a tumoral phenotype

Identification of the PTPN22 functional variant R620W as susceptibility genetic factor for giant cell arteritis

Objective: To analyse the role of the PTPN22 and CSK genes, previously associated with autoimmunity, in the predisposition and clinical phenotypes of giant cell arteritis (GCA). Methods: Our study population was composed of 911 patients diagnosed with biopsy-proven GCA and 8136 unaffected controls from a Spanish discovery cohort and three additional independent replication cohorts from Germany, Norway and the UK. Two functional PTPN22 polymorphisms (rs2476601/R620W and rs33996649/R263Q) and two variants of the CSK gene (rs1378942 and rs34933034) were genotyped using predesigned TaqMan assays. Results: The analysis of the discovery cohort provided evidence of association of PTPN22 rs2476601/R620W with GCA (PFDR=1.06E-04, OR=1.62, CI 95% 1.29 to 2.04). The association did not appear to follow a specific GCA subphenotype. No statistically significant differences between allele frequencies for the other PTPN22 and CSK genetic variants were evident either in the case/control or in stratified case analysis. To confirm the detected PTPN22 association, three replication cohorts were genotyped, and a consistent association between the PTPN22 rs2476601/R620W variant and GCA was evident in the overall meta-analysis (PMH=2.00E-06, OR=1.51, CI 95% 1.28 to 1.79). Conclusions: Our results suggest that the PTPN22 polymorphism rs2476601/R620W plays an important role in the genetic risk to GCA

Short-course combination treatment for experimental chronic Chagas disease.

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, affects millions of people in the Americas and across the world, leading to considerable morbidity and mortality. Current treatment options, benznidazole (BNZ) and nifurtimox, offer limited efficacy and often lead to adverse side effects because of long treatment durations. Better treatment options are therefore urgently required. Here, we describe a pyrrolopyrimidine series, identified through phenotypic screening, that offers an opportunity to improve on current treatments. In vitro cell-based washout assays demonstrate that compounds in the series are incapable of killing all parasites; however, combining these pyrrolopyrimidines with a subefficacious dose of BNZ can clear all parasites in vitro after 5 days. These findings were replicated in a clinically predictive in vivo model of chronic Chagas disease, where 5 days of treatment with the combination was sufficient to prevent parasite relapse. Comprehensive mechanism of action studies, supported by ligand-structure modeling, show that compounds from this pyrrolopyrimidine series inhibit the Qi active site of T. cruzi cytochrome b, part of the cytochrome bc1 complex of the electron transport chain. Knowledge of the molecular target enabled a cascade of assays to be assembled to evaluate selectivity over the human cytochrome b homolog. As a result, a highly selective and efficacious lead compound was identified. The combination of our lead compound with BNZ rapidly clears T. cruzi parasites, both in vitro and in vivo, and shows great potential to overcome key issues associated with currently available treatments