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  1. Home
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Browsing by Subject "Virus"

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    A new putative Caulimoviridae genus discovered through air metagenomics
    (American Society for Microbiology, 2018-10-11) Rastrojo, Alberto; Núñez, Andrés; Moreno, Diego A.; Alcamí, Antonio; Genética y Microbiología
    Members of the Caulimoviridae family are important plant pathogens. These circular double-stranded DNA viruses may integrate into the host genome, although this integration is not required for the viral replication cycle. Here, we describe three complete genomes belonging to a new putative Caulimoviridae genus discovered through air metagenomics.
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    Apoptosis of thymocytes in experimental African Swine Fever virus infection
    (Murcia : F. Hernández, 2004) Salguero, F.J.; Sánchez-Cordón, P.J.; Sierra, M. A.; Jover, A.; Núñez, A.; Gómez-Villamandos, J. C.
    This paper report on the lesions occurred in the thymus in experimental acute African swine fever (ASF). Twenty-one pigs were inoculated with the highly virulent ASF virus (ASFV) isolate Spain-70. Animals were slaughtered from 1 to 7 days post infection (dpi). Three animals with similar features were used as controls. Thymus samples were fixed in 10% buffered formalin solution for histological and immunohistochemical study and in 2.5% glutaraldehyde for ultrastructural examination. For immunohistochemical study, the avidin-biotin-peroxidase complex (ABC) technique was used to demonstrate viral protein 73 and porcine myeloid-histiocyte antigen SWC3 using specific monoclonal antibodies. Cell apoptosis was evaluated by the TUNEL assay. Blood samples were taken daily from all pigs and were used for leukocyte counts. The results of this study show a severe thymocyte apoptosis not related to the direct action of ASFV on these cells, but probably to a quantitative increase in macrophages in the thymus and their activation. A decrease in the percentage of blood lymphocytes was observed at the same time No significant vascular changes were observed in the study. With these results we suggest that ASFV infection of the thymus does not seem to play a critical role in the acute disease. Although severe apoptosis was observed, animals died because of the severe lesions found in the other organs.
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    Biomarkers of innate immunity and immunological susceptibility to viral Infection in patients with alcoholic cirrhosis
    (MDPI, 2024-02-01) Navarro Noguera, Elena; Collados Ros, Aurelia; Miguel Bolarín, Jose; Muro, Manuel; Legaz Pérez, Isabel; Ciencias Sociosanitarias
    Background: The harmful effect of alcohol on the immune system may be due to both a direct action of the alcohol or its metabolites on immune cells as an indirect action modifying the different mechanisms of intercellular interaction. The interplay between stimulatory (aKIR) and inhibitory (iKIR) natural killer (NK) cell receptors and their corresponding human leukocyte antigen (HLA) ligands influences the outcome of virus infection. The aim was to analyze the influence of the KIR/HLA pair genetic profile in male alcoholic cirrhosis (AC) patients with and without viral infections to find susceptibility biomarkers that can help establish the risks and prevent viral infections. Methods: A total of 281 male AC patients were analyzed. The sociodemographic characteristics, viral hepatitis C (HCV), hepatitis B (HBV), and cytomegalovirus (CMV) infections were analyzed. Genomic DNA was extracted, and genetic the KIR/HLA profiles were investigated. A total of 6 KIR genes and their corresponding ligands (HLA-C) were analyzed. Patients were grouped into two groups: with and without associated viral infection. Results: A statistically significant increase in the combination of KIR2DL2+/C1C1 was observed in male AC patients with viral infection compared to those without viral infection (45.9% vs. 24.5%, p = 0.021). The analysis of KIR2DL3+/C1+ showed a high frequency comparing healthy controls and male AC patients without virus infection (85% vs. 76.4%; p = 0.026). The analysis of KIR2DL3+/C2C2 frequency showed a statistically significant increase comparing male AC patients without viral infection and healthy controls (23.6% vs. 15%; p = 0.026). Conclusions: The genetic KIR2DL2+/C2C2 profiles may play a significant role in determining the vulnerability of male AC patients to viral infections, providing valuable insights for future research and potential therapeutic interventions.
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    Infección por el virus del papiloma humano en mujeres con lesión intraepitelial cervical : aspectos virológicos y clinicopatológicos
    (2014-02-10) Moreno Docón, Antonio; Segovia Hernández, Manuel; Departamento de Genética y Microbiología
    Objetivos: En la actualidad, se conoce bien la relación que existe entre la infección por el virus del papiloma humano (VPH) y las lesiones precursoras del cáncer cervical. El principal objetivo de este estudio fue conocer la prevalencia general de la infección por VPH y estudiar los genotipos más frecuentes en mujeres con lesión intraepitelial cervical en la región de Murcia. Por otra parte, se ha evaluado la utilidad de la detección de VPH como marcador pronóstico de enfermedad residual en mujeres tratadas por lesión cervical de alto grado (CIN2+). Material y métodos: Se estudiaron muestras clínicas procedentes de 3.752 pacientes que tras una exploración ginecológica presentaron un resultado citológico anormal. Este estudio se realizó durante un periodo de 5 años (2008-2012), en un contexto de cribado asistencial oportunista, al no existir un modelo de cribado poblacional en nuestra comunidad. La detección de ADN del VPH se realizó mediante la técnica de captura de híbridos, mientras que la detección y tipado de VPH se realizó mediante una técnica de PCR microarray. Resultados: En nuestro estudio, encontramos que la tasa de detección de VPH fue del 53,5% en ASCUS, 73,2% en LSIL y 78,9% en HSIL. Los cuatro genotipos más frecuentes encontrados fueron los tipos 16 (31,1%), 31 (13,5%), 51 (13,4%) y el 53 (11,8%). La prevalencia del VPH 16 aumentó significativamente con el grado de la lesión citológica (p < 0,0001). El VPH 18 se detectó con una baja frecuencia (5,7%), datos en concordancia con otros estudios publicados en nuestro país. No encontramos diferencias significativas entre el número de tipos de VPH y el grado de severidad de la lesión cervical. La introducción del test de VPH en el seguimiento de mujeres tratadas mediante escisión con asa de diatermia, dada su elevada sensibilidad (93,7%) y valor predictivo negativo (97,3%), permite que estas mujeres pueden realizar su seguimiento mediante un protocolo de cribado convencional cada 3-5 años. ABSTRACT Objetives: At present, the relationship between human papillomavirus (HPV) infection and cervical preneoplasic lesions is well known. The aim of this study was to determine the overall prevalence of HPV infection, and study the most frequent genotypes in women with cervical intraepithelial lesion in the region of Murcia. Moreover, we evaluated the usefulness of HPV detection as a prognostic marker of residual disease in women treated for high-grade cervical lesions (CIN2+). Material and Methods: We studied 3.752 clinical samples from patients after gynecological examination which showed an abnormal cytology result. This study was conducted over a period of five years (2008-2012), in a context of opportunistic screening in the absence of a model for population screening in our community. Detection of HPV DNA was performed by hybrid capture technique, whereas HPV detection and typing was performed by microarray-PCR. Results : In our study we found that HPV detection rate was 53.5% in ASCUS , 73.2 % in LSIL and 78.9 % in HSIL . The four most common genotypes found were types 16 ( 31.1 % ) , 31 ( 13.5 % ) , 51 ( 13.4%) and 53 ( 11.8 % ) . The prevalence of HPV 16 increased significantly with the degree of cervical lesion (p <0.0001). The HPV 18 was detected with a low frequency (5.7 %), according to data published in other studies in our country. There are no significant differences between the number of HPV types and degree of severity of the cervical lesion. The introduction of HPV testing in the monitoring of women treated by diathermy loop excision , given its high sensitivity (93.7 % ) and negative predictive value ( 97.3 % ) , allows these women to follow it up with a conventional screening protocol every 3-5 years.
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    Marinomonas mediterranea synthesizes an R-type bacteriocin
    (American Society for Microbiology, 2024-01-03) Lucas-Elío, Patricia; ElAlami, Tarik; Martínez, Alicia; Sanchez-Amat, Antonio; Genética y Microbiología; Zoología y Antropología Física
    Prophages integrated into bacterial genomes can become cryptic or defective prophages, which may evolve to provide various traits to bacterial cells. Previous research on Marinomonas mediterranea MMB-1 demonstrated the production of defective particles. In this study, an analysis of the genomes of three different strains (MMB-1, MMB-2, and MMB-3) revealed the presence of a region named MEDPRO1, spanning approximately 52 kb, coding for a defective prophage in strains MMB-1 and MMB-2. This prophage seems to have been lost in strain MMB-3, possibly due to the presence of spacers recognizing this region in an I-F CRISPR array in this strain. However, all three strains produce remarkably similar defective particles. Using strain MMB-1 as a model, mass spectrometry analyses indicated that the structural proteins of the defective particles are encoded by a second defective prophage situated within the MEDPRO2 region, spanning approximately 13 kb. This finding was further validated through the deletion of this second defective prophage. Genomic region analyses and the detection of antimicrobial activity of the defective prophage against other Marinomonas species suggest that it is an R-type bacteriocin. Marinomonas mediterranea synthesizes antimicrobial proteins with lysine oxidase activity, and the synthesis of an R-type bacteriocin constitutes an additional mechanism in microbial competition for the colonization of habitats such as the surface of marine plants.
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    Turbot TNFα gene: molecular characterization and biological activity of the recombinant protein
    (Elsevier, 2006-04-17) Ordas, M. C.; Costa, Maria del Mar; Lopez-Castejón, Gloria; Meseguer Peñalver, J.; Figueras, Antonio; Novoa, Beatriz; Mulero Méndez, Victoriano Francisco; Roca Soler, Francisco José; Bioquímica y Biología Molecular B e Inmunología
    The tumor necrosis factor (TNF) superfamily is composed by several proteins with similar structure and functions. One of the main representatives of this family is TNF-alpha (TNFα), a proinflammatory cytokine which is produced by different immune cells and presents a wide variety of activities. Using the RACE technique, we have cloned and sequenced the turbot TNF cDNA. The analysis of its sequence showed several conserved motifs characteristic of members of the TNFα family. A phylogenetic tree constructed with different TNFs of fish and mammals grouped our sequence within the fish TNFα cluster. Therefore, the turbot TNF here studied was identified as TNFα. The complete TNFα gene was obtained by gene walking, and, similarly to the other known fish TNFα genes, presented three introns and four exons. A PCR was designed to study the turbot TNFα expression in vivo using as stimulus the bacteria Vibrio pelagius strain Hq222 and virus VHSV. The expression of the cytokine happened early after injection, and it was dependent on the pathogen injected and organ analyzed. Virus induced a higher TNFα expression, but this response was shorter in time than that induced by bacteria. In addition, TNFα expression was in general higher in kidney than in liver, as expected since the former is the haematopoietic organ of fish. The turbot recombinant TNFα (rTNFα) was obtained by IPTG induction of bacteria transformed with the pET15b-TNFα construct, and it was purified in native conditions. The recombinant protein was approximately 20 kDa in size, and its biological activity was assessed in vitro. No effect of the rTNFα neither alone nor in combination with LPS was observed on the chemiluminescence activity of turbot macrophages at any time tested. However, NO production was enhanced by the recombinant protein alone or with LPS 72 h after the addition of the treatments. Finally, turbot rTNFα was able to recruit and activate inflammatory cells when injected in gilthead seabream, although to a lesser extent than gilthead seabream rTNFα.
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    Ultrastructural pathology of the bone marrow in pigs inoculated with a moderately virulent strain (DR'78) of African swine fever virus
    (Murcia : F. Hernández, 1998) Bautista, María José; Gómez-Villamandos, J. C.; Carrasco, L.; Ruíz-Villamor, E.; Salguero, F.J.; Sierra, M. A.
    Interpretation of changes in bone marrow during infectious processes is quite complex. This paper reports bone marrow lesions observed in pigs inoculated with a moderately virulent ASF virus strain and studies their relationship to the pathogenesis of the disease. In this work, we have carried out the structural and ultrastructural study of the bone marrow of 14 Large White X Landrace pigs that were inoculatd by the intramuscular route with 105 50% hemodsorbing doses (HADSO) of the Dominican Republic178 ASF virus strain. The inoculated pigs were killed at 3, 5, 7, 9, 11, 13, 15 and 17 days postinjection. Analysis of cells and structures belonging to the two main bone-marrow compartments, the hematopoietic cells and the hematopoietic microenvironment, showed that after inoculation with a moderately virulent strain, the most significant changes occurred in macrophages and megakaryocytes, consisting in virus replication in these cell populations and apoptosis of megakaryocytes, related with the sudden and transitory thrombocytopenia detected in the subacute ASF.

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