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5 Pág., Project VIROPLANT, European Union's Horizon 2020 Research and Innovation Programme Grant/Award Number: 773567

Resumen Este artículo se propuso un objetivo: visibilizar la forma como el cuidado se activa para ofrecer acompañamiento al dolor desde las experiencias de la Asociación de Mujeres Afro por la Paz, quienes desarrollan estrategias de agenciamiento político que contribuyen a dar sentido a sus vivencias colectivas y a asegurar el valor de su memoria y la construcción de paz conforme a sus prácticas ancestrales como víctimas-sobrevivientes. De esta manera se reconoce la importancia de una plataforma política mientras se transmiten elementos claves para el cuidado, como son: solidaridad, confianza y empatía, con la finalidad de movilizar nuevas ciudadanías en favor de la defensa de la vida y los derechos humanos. El artículo es el resultado de un proceso de investigación con enfoque narrativo como método, y que comprende experiencias humanas en claves simbólicas cuya narración crea un marco de referencia donde se valora reflexivamente la práctica terapéutica del cuidado para reparar y acompañar el dolor vivido por las mujeres durante la guerra. Abstract The objective of the article is to make visible the way care is activated around the accompaniment of pain from the experiences of the Association of Afro Women for Peace, which supports strategies of political agency, which provide a sense of their collective experiences and thus recognize the importance of a political platform that ensures the value of its memory and the construction of peace subject to its ancestral practices as victims-survivors which transmit key elements for the care of solidarity, trust and empathy with the purpose of mobilize new citizenships committed to the defense of life and human rights. This article is the result of a research process of which I approach the narrative approach as a method which consists of human lessons to give meaning and meaning to the behaviors of the subjects through the story that involves symbolic keys in which the narrative is a frame of reference and seeks to reflectively contribute to assess care as to accompany the pain of the women lived in the war. Resumo O objetivo do artigo é tornar visível a forma como o cuidado é acionado em torno do acompanhamento da dor a partir das experiências da Associação de Mulheres Afro-Americanas pela Paz, que apóia estratégias de atuação política, que proporcionam um senso de suas experiências coletivas. e assim reconhecer a importância de uma plataforma política que assegure o valor de sua memória e a construção da paz sujeita às suas práticas ancestrais como vítimassobreviventes que transmitem elementos-chave para o cuidado da solidariedade, confiança e empatia com o propósito de mobilizar novas cidadanias comprometidas com a defesa da vida e dos direitos humanos. Este artigo é o resultado de um processo de pesquisa que abordou a abordagem narrativa como um método que consiste em aulas humanos para dar sentido e significado aos comportamentos de indivíduos através da história envolvendo chaves simbólicas em que a narrativa é um quadro de referência e procura contribuir reflexivamente para avaliar o cuidado de acompanhar a dor das mulheres vividas na guerra.

CAR-T-cell therapy against MM currently shows promising results, but usually with serious toxicities. CAR-NK cells may exert less toxicity when redirected against resistant myeloma cells. CARs can be designed through the use of receptors, such as NKG2D, which recognizes a wide range of ligands to provide broad target specificity. Here, we test this approach by analyzing the antitumor activity of activated and expanded NK cells (NKAE) and CD45RA− T cells from MM patients that were engineered to express an NKG2D-based CAR. NKAE cells were cultured with irradiated Clone9.mbIL21 cells. Then, cells were transduced with an NKG2D-4-1BB-CD3z-CAR. CAR-NKAE cells exhibited no evidence of genetic abnormalities. Although memory T cells were more stably transduced, CAR-NKAE cells exhibited greater in vitro cytotoxicity against MM cells, while showing minimal activity against healthy cells. In vivo, CAR-NKAE cells mediated highly efficient abrogation of MM growth, and 25% of the treated mice remained disease free. Overall, these results demonstrate that it is feasible to modify autologous NKAE cells from MM patients to safely express a NKG2D-CAR. Additionally, autologous CAR-NKAE cells display enhanced antimyeloma activity demonstrating that they could be an effective strategy against MM supporting the development of NKG2D-CAR-NK-cell therapy for MM., This study was supported by a grant from the Spanish Society for Hematology and Hemotherapy to Alejandra Leivas, the CRIS Foundation to Beat Cancer and the Instituto de Salud Carlos III (PI18/01519).

Se describe el patrón de inervación del miembro posterior con el establecimiento de los plexos lumbosacros y los nervios femoral, ciático, tibial, fibular y plantares en el embrión de rata (Rattus norvergicus). La metodología utilizada permitió una mejor visualización de este fenómeno biológico y su relación con otras estructuras corporales como cartílago, ganglios sensoriales y médula espinal ya que todas ellas sufren cambios que se presentan tanto en el espacio como en el tiempo. Además, el modelo 3D generado será de gran utilidad para posteriores investigaciones en este campo, al igual que de herramienta docente para ilustrar de manera sencilla y adecuada este complejo fenómeno.

ZnO nanoparticles (NPs) are arising as promising novel antibiotics toward device-related infections. The surface functionalization of Zn, a novel resorbable biomaterial, with ZnO NPs could present an effective solution to overcome such a threat. In this sense, the antibacterial and antibiofilm activity of nano- and microsized ZnO coatings was studied against clinically relevant bacteria, methicillin resistant Staphylococcus aureus (MRSA). The bacterial viability of planktonic and biofilm cells together with the corresponding biofilm structures revealed that only the nanosized ZnO coating had an antibiofilm effect. To elucidate this effect, a novel approach was taken: preconditioning of bacteria with this ZnO coating followed by exposure to subinhibitory concentrations of antibiotics with well-known modes of actions. This approached revealed (i) a decreased biofilm formation in combination with gentamycin, targeting protein synthesis, and (ii) an increased biofilm formation in the presence of rifampicin and vancomycin, acting on RNA and cell wall biosynthesis, respectively. The increased bacteria resistance to these two antibiotics gave new insights into the antibiofilm effect of this nanosized ZnO coating. The synergistic effect observed for gentamycin opened new perspectives for the design of effective solutions against implant-related infections. During the in vitro degradation of this nanosized ZnO-coated Zn, a specific corrosion product, hopeite [Zn

Background: Acute myeloid leukemia (AML) is a hematological malignancy with a very low overall survival. Among the new treatment modalities, chimeric antigen receptor (CAR) therapy is showing promising results in other hematological malignancies. Since AML exhibits high heterogeneity and does not have specific differential antigens of the hematopoietic stem cell, using NKG2D-CAR cells could be an appropriate therapeutic strategy against AML. NKG2D receptor has a wide range of specific tumor cell ligands (MICA, MICB, ULBP-1, ULBP-2 and ULBP-3) which are expressed in more than 80% of all tumors. For this reason, the objective of this work was to evaluate the anti-tumor activity of activated and expanded natural killer cells (NKAE) and T cells expressing an NKG2D CAR. Methods: T cells and NK cells were isolated from the healthy donor´s peripheral blood mononuclear cells ring (n = 5) by immunomagnetic depletion. NKAE cells were obtained by co-culture with subletally irradiated CSTX002 cells. The purified NKAEs and T cells were transduced with an NKG2D CAR with 4-1BB and CD3z signaling domains. The efficiency of transduction was evaluated by flow cytometry detecting NKG2D expression. Also, the immunoprofiling of surface molecules and the cytotoxicity against primary blasts of AML, as well as the expression of NKG2D ligands in tumor cells (AML cell lines and primary blasts) were analyzed by flow cytometry. The cytotoxicity of untransduced NKAE, CAR-NKAE cells, untransduced T cells and CAR-T cells was evaluated by 4 hour europium release assay. Toxicity on healthy tissue (healthy lung cells and PBMCs from third party) was analyzed in the same way. The safety of NKG2D-CAR transduced cells was evaluated using CGH arrays to detect chromosomal abnormalities. Results: Both the AML cell lines and primary blasts from AML patients showed expression of the MICA/B and ULBPs-1 to 3 ligands. NKG2D ligands expression was highly variable from a cell line to another. However, all the cell lines and samples showed high expression of at least 1 of the 5 analyzed ligands. Four hour europium release assays revealed that untransduced T cells had higher cytotoxicity than untransduced NKAE cells at the same ratio (32: 1) against both OCI-AML-3 cytarabine-resistant cell line (52.7% ± 14.2% vs. 32.5% ± 7.2%) as to the cytarabine-sensitive cell line (56.42% ± 5.6% vs. 50.14 % ± 5.9%). T cells showed a better transduction efficiency than NKAE cells at a multiplicity of infection (MOI) of 5. It was observed that both the CAR-NKAE cells and the CAR-T cells had higher cytotoxicity against these two lines (OCI-AML-3R and OCI-AML-3S) after being transduced with our NKG2D-CAR. However, the antitumor activity of CAR-T cells always remained superior to that of the CAR-NKAE for both OCI-AML-3R (54.26% ± 3.8% vs. 35.3% ± 6.7%) and for OCI-AML-3S (63.36% ± 3.5% vs. 54.3% ± 3.7%) cell lines, with a greater difference compared to drug resistant cells. The antitumor activity of CAR-T cells and CAR-NKAE cells was always superior on the sensitive cell line. The antitumor activity of NKG2D CAR-T cells was evaluated against primary blasts from AML patients (n = 4), observing a nearly complete destruction of the blasts after 24 hours, at a very low target: effector ratio of 4:1. CAR-T cells populations were analyzed by flow cytometry and we found that CAR-T cells were highly positive to CD45RO and reduced expression of CD45RA, on the other hand, they showed high IL-2 production exhibiting a central memory phenotype. Slight toxicity was observed against lung cells in both cell types. However, CAR-T cells exhibited some toxicity on third party PBMCs being null in the case of CAR-NKAE. CGH arrays studies showed no variation in the copy number, resulting from the introduction of the CAR, of genes which their variation has been associated with chromosomal instability. No significant changes associated with transduction process were observed in the surface phenotype of CAR-NKAE cells or CAR-T cells. Conclusions: We have demonstrated that AML cells could be target with an NKG2D-CAR. Primary NKAE cells and T cells can be transduced with an NKG2D-CAR at a very low MOI to enhance their antileukemic activity. CD3+ CAR-T cells are more effective than CAR-NKAE cells. Moreover, CAR-T cells were able to completely destroy AML blasts. Although further studies are needed, these results show the potential of NKG2D-CAR T and NK cell therapy in AML. Disclosures Rio: Rocket Pharmaceuticals: Equity Ownership, Patents & Royalties, Research Funding. Lee:Kiadis Pharma: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding.

El Grupo de lnvestigación PACA de la Universidad Surcolombiana ha liderado el proyecto "lndagación sistemática sobre los factores de permanencia y graduación estudiantiles en la Uni­versidad Surcolambiana" el cual hace parte del Convenio 619 de octubre de 2012 celebrado con el Ministerio de Educación Nacional. En ese sentido, desde octubre del año 2012 el Grupo de Investigación PACA encabezado poir el docente investigador Nelson Ernesto López Jiménez y las Magíster Karol Stefanía Amaya García y Laura Cordoba Muñoz han desarrollado las actividades que se definieron en el mar­co de la convocatoria y que responden a cada uno de los componentes que han permitido abordar la problemática de la permanencia y del abandono estudiantil en la Universidad Surcolombiana.

La Primera lnfancia, comprendida entre los cero y los seis años, es una etapa en la que tanto los niños como las niñas requieren estimulación, atención y tratamiento especiales que les permita desarrollarse biológica, psicológica, cultural y socialmente. Diversos estudios reali­zados desde distintas disciplinas han comprobado con argumentos sólidos que desde su concepción los niños cuentan con una serie de capacidades físicas, psicológicas, emociona­les y sociales que facilitan el aprendizaje, la socialización y la comunicación.

El Grupo de investigación PACA - Categoría A de Colciencias ha adelan­tado investigaciones en torno al fenómeno de la permanencia escolar", en ellos se ha identificado una problemática relacionada con el estado particular entre el abandono y la perrnanencia", la cual se ha denomi­nado: migración escolar. Este fenómeno es entendido como el cambio de programa académico que realiza el estudiante universitario en la misma institución, después de haber cursado un número determinado de semestres.

Endothelial cells (EC) are able to actively control vascular permeability, coagulation, blood pressure and angiogenesis. Most recently, a role for endothelial cells in the immune response has been described. Therefore, the endothelium has a dual role controlling homeostasis but also being the first line for host defence and tissue damage repair thanks to its ability to mount an inflammatory response. Endothelial cells have been shown to express pattern-recognition receptors (PRR) including Toll-like receptors (TLR) that are activated in response to stimuli within the bloodstream including pathogens and damage signals. TLRs are strategic mediators of the immune response in endothelial cells but they also regulate the angiogenic process critical for tissue repair. Nevertheless, endothelial activation and angiogenesis can contribute to some pathologies. Thus, inappropriate endothelial activation, also known as endothelial dysfunction, through TLRs contributes to tissue damage during autoimmune and inflammatory diseases such as atherosclerosis, hypertension, ischemia and diabetes associated cardiovascular diseases. Also TLR induced angiogenesis is required for the growth of some tumors, atherosclerosis and rheumatoid arthritis, among others. In this review we discuss the importance of various TLRs in modulating the activation of endothelial cells and their importance in immunity to infection and vascular disease as well as their potential as therapeutic targets.

Hepatitis E virus (HEV) is an important but extremely understudied human pathogen, and the mechanisms of HEV replication and pathogenesis are largely unknown. We previously identified an attenuated genotype 3 HEV mutant (pSHEV-1) containing three unique amino acid mutations (F51L, T59A, and S390L) in the capsid protein. To determine the role of each of these mutations, we constructed three HEV single mutants (rF51L, rT59A, and rS390L) which were all found to be replication competent in Huh7 liver cells. To determine the pathogenicities of the mutants, we utilized the specific-pathogen-free (SPF) pig model for HEV and a unique inoculation procedure that bypasses the need for propagating infectious HEV in vitro . A total of 60 pigs were intrahepatically inoculated, via an ultrasound-guided technique, with in vitro -transcribed full-length capped RNA transcripts from the infectious clones of each single mutant, the pSHEV-1 triple mutant, wild-type pSHEV-3, or phosphate-buffered saline (PBS) buffer ( n = 10). The results showed that the F51L mutation partially contributed to virus attenuation, whereas the T59A and S390L mutations resulted in more drastic attenuation of HEV in pigs, as evidenced by a significantly lower incidence of viremia, a delayed appearance and shorter duration of fecal virus shedding and viremia, and lower viral loads in liver, bile, and intestinal content collected at three different necropsy times. The results indicate that the three mutations in the capsid protein collectively contribute to HEV attenuation. This study has important implications for developing a modified live-attenuated vaccine against HEV.

Genotypes 1 and 2 hepatitis E virus (HEV) infect only humans whereas genotypes 3 and 4 HEV infect both humans and pigs. To evaluate the mechanism of cross-species HEV infection between humans and swine, in this study we constructed five intergenotypic chimeric viruses and tested for their infectivity in vitro and in pigs. We demonstrated that chimeric viruses containing the ORF2 capsid gene either alone or in combination with its adjacent 5' junction region (JR) and 3' noncoding region (NCR) from a genotype 4 human HEV in the backbone of a genotype 3 swine HEV are replication-competent in Huh7 cells and infectious in HepG2/C3A cells and in pigs, and thus supporting the hypothesis that genotypes 3 and 4 human HEV are of swine origin. However, chimeric viruses containing the JR+ORF2+3' NCR of genotypes 3 or 4 HEV in the backbone of genotype 1 human HEV failed to infect pigs, suggesting that other genomic regions such as 5' NCR and ORF1 may also be involved in HEV cross-species infection. The results from this study provide the first experimental evidence of the exchangeability of the capsid gene between genotype 3 swine HEV and genotype 4 human HEV, and have important implications for understanding the mechanism of HEV cross-species infection.

Lo curricular en el contexto educativo se convierte en la expresión de la complejidad, que caracteriza los procesos de formación profesional en los diferentes niveles de la educación superior en nuestro país. Una de las características que más han generado la reflexión académica en materia curricular es la que hace referencia a la pertenencia social y pertinencia académica de los diferentes programas que actualmente constituyen el universo de la educación superior en Colombia y de la educación formal en general. Si se asume que todos los programas académicos se sustentan en necesidades que los justifican, es conveniente problematizar la manera como han sido diseñados los componentes que forman parte de las diferentes estructuras curriculares. No son nuevos los cuestionamientos que se les hacen a los perfiles académicos que se constituyen en el referente por excelencia en la mayoría de programas existentes hoy en día en la educación superior de nuestro país.

Porcine circovirus type 1 (PCV1), originally isolated as a contaminant of PK-15 cells, is nonpathogenic, whereas porcine circovirus type 2 (PCV2) causes an economically important disease in pigs. To determine the factors affecting virus replication, we constructed chimeric viruses by swapping open reading frame 1 (ORF1) ( rep ) or the origin of replication (Ori) between PCV1 and PCV2 and compared the replication efficiencies of the chimeric viruses in PK-15 cells. The results showed that the replication factors of PCV1 and PCV2 are fully exchangeable and, most importantly, that both the Ori and rep of PCV1 enhance the virus replication efficiencies of the chimeric viruses with the PCV2 backbone.

Hepatitis E virus (HEV) is a single-strand positive-sense RNA virus in the family Hepeviridae . The disease caused by HEV, hepatitis E, is an important public health problem in developing countries of Asia and Africa and is also endemic in many industrialized countries, including the United States. HEV has been identified from several other animal species in addition to humans, including the pig, chicken, mongoose, deer, rabbit, ferret, bat, and fish. Here we report the complete genome sequence of the first strain of HEV from rabbits in the United States. Sequence and phylogenetic analyses revealed that the U.S. rabbit HEV is a distant member of the zoonotic genotype 3 HEV, thus raising a concern for potential zoonotic human infection. A unique 90-nucleotide insertion within the X domain of the ORF1 was identified in the rabbit HEV, and this insertion may play a role in the species tropism of HEV.

Hepatitis E virus (HEV) is an important but extremely understudied human pathogen. Genotypes 1 and 2 are restricted to humans, whereas genotypes 3 and 4 are zoonotic, infecting both humans and pigs. This report describes, for the first time, the successful rescue of infectious HEV in vitro and in vivo from cloned cDNA of a genotype 4 human HEV (strain TW6196E). The complete genomic sequence of the TW6196E virus was determined and a full-length cDNA clone (pHEV-4TW) was assembled. Capped RNA transcripts from the pHEV-4TW clone were replication competent in Huh7 cells and infectious in HepG2/C3A cells. Pigs inoculated intrahepatically with capped RNA transcripts from pHEV-4TW developed an active infection, as evidenced by faecal virus shedding and seroconversion, indicating the successful rescue of infectious genotype 4 HEV and cross-species infection of pigs by a genotype 4 human HEV. To demonstrate the utility of the genotype 4 HEV infectious clone and to evaluate the potential viral determinant(s) for species tropism, four intergenotypic chimeric clones were constructed by swapping various genomic regions between genotypes 1 and 4, and genotypes 1 and 3. All four chimeric clones were replication competent in Huh7 cells, but only the two chimeras with sequences swapped between genotypes 1 and 4 human HEVs produced viruses capable of infecting HepG2/C3A cells. None of the four chimeras was able to establish a robust infection in pigs. The availability of a genotype 4 HEV infectious clone affords an opportunity to delineate the molecular mechanisms of HEV cross-species infection in the future.

Hepatitis E virus (HEV), the causative agent of hepatitis E, is primarily transmitted via the fecal-oral route through contaminated water supplies, although many sporadic cases of hepatitis E are transmitted zoonotically via direct contact with infected animals or consumption of contaminated animal meats. Genotypes 3 and 4 HEV are zoonotic and infect humans and other animal species, whereas genotypes 1 and 2 HEV are restricted to humans. There exists a single serotype of HEV, although the cross-protective ability among the animal HEV strains is unknown. Thus, in this study we expressed and characterized N-terminal truncated ORF2 capsid antigens derived from swine, rat, and avian HEV strains and evaluated their cross-protective ability in a pig challenge model. Thirty, specific-pathogen-free, pigs were divided into 5 groups of 6 pigs each, and each group of pigs were vaccinated with 200 µg of swine HEV, rat HEV, or avian HEV ORF2 antigen or PBS buffer (2 groups) as positive and negative control groups. After a booster dose immunization at 2 weeks post-vaccination, the vaccinated animals all seroconverted to IgG anti-HEV. At 4 weeks post-vaccination, the animals were intravenously challenged with a genotype 3 mammalian HEV, and necropsied at 4 weeks post-challenge. Viremia, fecal virus shedding, and liver histological lesions were compared to assess the protective and cross-protective abilities of these antigens against HEV challenge in pigs. The results indicated that pigs vaccinated with truncated recombinant capsid antigens derived from three animal strains of HEV induced a strong IgG anti-HEV response in vaccinated pigs, but these antigens confer only partial cross-protection against a genotype 3 mammalian HEV. The results have important implications for the efficacy of current vaccines and for future vaccine development, especially against the novel zoonotic animal strains of HEV.

Hepatitis E virus (HEV) is an important human pathogen. In addition to humans, HEV has also been identified in pig, chicken, mongoose, deer, rat, rabbit and fish. There are four recognized and two putative genotypes of mammalian HEV. Genotypes 1 and 2 are restricted to humans, while genotypes 3 and 4 are zoonotic. The recently identified rabbit HEV is a distant member of genotype 3. Here, we first expressed and purified the recombinant capsid protein of rabbit HEV and showed that the capsid protein of rabbit HEV cross-reacted with antibodies raised against avian, rat, swine and human HEV. Conversely, we showed that antibodies against rabbit HEV cross-reacted with capsid proteins derived from chicken, rat, swine and human HEV. Since pigs are the natural host of genotype 3 HEV, we then determined if rabbit HEV infects pigs. Twenty pigs were divided into five groups of four each and intravenously inoculated with PBS, US rabbit HEV, Chinese rabbit HEV, US rat HEV and swine HEV, respectively. Results showed that only half of the pigs inoculated with rabbit HEV had low levels of viraemia and faecal virus shedding, indicative of active but not robust HEV infection. Infection of pigs by rabbit HEV was further verified by transmission of the virus recovered from pig faeces to naïve rabbits. Pigs inoculated with rat HEV showed no evidence of infection. Preliminary results suggest that rabbit HEV is antigenically related to other HEV strains and infects pigs and that rat HEV failed to infect pigs.

The RNA genome of the hepatitis E virus (HEV) contains a hypervariable region (HVR) in ORF1 that tolerates small deletions with respect to infectivity. To further investigate the role of the HVR in HEV replication, we constructed a panel of mutants with overlapping deletions in the N-terminal, central, and C-terminal regions of the HVR by using a genotype 1 human HEV luciferase replicon and analyzed the effects of deletions on viral RNA replication in Huh7 cells. We found that the replication levels of the HVR deletion mutants were markedly reduced in Huh7 cells, suggesting a role of the HVR in viral replication efficiency. To further verify the results, we constructed HVR deletion mutants by using a genetically divergent, nonmammalian avian HEV, and similar effects on viral replication efficiency were observed when the avian HEV mutants were tested in LMH cells. Furthermore, the impact of complete HVR deletion on virus infectivity was tested in chickens, using an avian HEV mutant with a complete HVR deletion. Although the deletion mutant was still replication competent in LMH cells, the complete HVR deletion resulted in a loss of avian HEV infectivity in chickens. Since the HVR exhibits extensive variations in sequence and length among different HEV genotypes, we further examined the interchangeability of HVRs and demonstrated that HVR sequences are functionally exchangeable between HEV genotypes with regard to viral replication and infectivity in vitro , although genotype-specific HVR differences in replication efficiency were observed. The results showed that although the HVR tolerates small deletions with regard to infectivity, it may interact with viral and host factors to modulate the efficiency of HEV replication.

SUMMARYWest Nile virus (WNV) was probably introduced in southern and northern Mexico from the USA in two independent events. Since then, WNV activity has been reported in several Mexican states bordering the USA and the Gulf of Mexico, but disease manifestations seen there in humans and equids are quite different to those observed in the USA. We have analysed WNV seroprevalence in asymptomatic, unvaccinated equids from two Mexican states where no data had been previously recorded. WNV IgG antibodies were detected in 31·6% (91/288) of equine sera from Chiapas and Puebla states (53·3% and 8·0%, respectively). Analysis by plaque reduction neutralization test (PRNT) showed good specificity (99·4%) and sensitivity (84·9%) with the ELISA results. Further analyses to detect antibodies against three different flaviviruses (WNV, St Louis encephalitis virus, Ilheus virus) by haemagglutination inhibition (HI) tests on a subset of 138 samples showed that 53% of the 83 HI-positive samples showed specific reaction to WNV. These data suggest continuous expansion of WNV through Mexico.

West Nile fever outbreaks in the USA have caused over 700 human deaths, primarily due to neurological disease. The usual transmission route of West Nile virus (WNV) involves mosquito bites; however, alternative routes, including intrauterine infection, have also been reported. Here, the pathogenicity of WNV in mice during gestation has been investigated. An extremely high mortality rate was observed in pregnant mice (98 %, 60/61) compared with non-pregnant mice (52 %, 28/53; P

Animal enteroviruses shed in the feces of infected animals are likely environmental contaminants and thus can be used as indicators of animal fecal pollution. Previous work has demonstrated that bovine enterovirus (BEV) present in bovine feces contaminates waters adjacent to cattle herds and that BEV-like sequences are also present in shellfish and in deer feces from the same geographical area. However, little information is available about the prevalence, molecular epidemiology, and genomic sequence variation of BEV field isolates. Here we describe an optimized highly sensitive real-time reverse transcription-PCR method to detect BEV RNA in biological and environmental samples. A combination of the amplification procedure with a previously described filtration step with electropositive filters allowed us to detect up to 12 BEV RNA molecules per ml of water. The feasibility of using the method to detect BEV in surface waters at a high risk of fecal pollution was confirmed after analysis of water samples obtained from different sources. The method was also used to study the prevalence of BEV in different cattle herds around Spain, and the results revealed that 78% (78 of 100) of the fecal samples were BEV positive. BEV-like sequences were also detected in feces from sheep, goats, and horses. Nucleotide sequence analyses showed that BEV isolates are quite heterogeneous and suggested the presence of species-specific BEV-like variants. Detection of BEV-like sequences may help in the differentiation and characterization of animal sources of contamination. Copyright © 2005, American Society for Microbiology. All Rights Reserved.