Isolation and characterization of immortalized porcine aortic endothelial cell lines   总被引：6，自引：0，他引：6  

Carrillo A  Chamorro S  Rodríguez-Gago M  Alvarez B  Molina MJ  Rodríguez-Barbosa JI  Sánchez A  Ramírez P  Muñoz A  Domínguez J  Parrilla P  Yélamos J 《Veterinary immunology and immunopathology》2002,89(1-2):91-98

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).  相似文献   

Specific polymerase chain reaction for differential diagnosis of Dirofilaria immitis and Dipetalonema reconditum using primers derived from internal transcribed spacer region 2 (ITS2)   总被引：2，自引：0，他引：2  

Mar PH  Yang IC  Chang GN  Fei AC 《Veterinary parasitology》2002,106(3):243-252

Both Dirofilaria immiti and Dipetalonema reconditum may be found in blood of infected dogs but it is not easy to distinguish D. immitis from D. reconditum in morphology. We cloned and sequenced the contiguous internal transcribed spacer (ITS) region, ITS1-5.8S-ITS2, of these two different parasites and published on GenBank as AF217800 for D. immiti and AF217801 for D. reconditum in this study. We designed two pairs of specific primers derived from ITS2 being used for polymerase chain reaction (PCR). The amplicons of ITS2 from D. immiti and D. reconditum are 302 and 348bp, respectively. Moreover, the limitation for amplifying ITS2 gene using this PCR demonstrated that 1 x 10(-2) microfilaria of each species of parasite smashed or even with mixed samples could be detected and the PCR products were predicted as the same as that described above. Thus, D. immiti and D. reconditum could be differentially diagnosed by this specific PCR. Seventeen clinical cases were evaluated and all of them were correctly identified. In this study, ITS1-5.8S-ITS2 of D. immiti or D. reconditum were the first time sequenced and analyzed. No significant similarity of ITS1 and ITS2 between D. immiti and D. reconditum could be observed.  相似文献   

Assessment of relevant factors and relationships concerning human dermal exposure to pesticides in greenhouse applications     

Martínez Vidal JL  Egea González FJ  Garrido Frenich A  Martínez Galera M  Aguilera PA  López Carrique E 《Pest management science》2002,58(8):784-790

Principal component analysis (PCA) was applied to the gas chromatographic data obtained from 23 different greenhouse trials. This was used to establish which factors, including application technique (very small, small, medium and large drop-size), crop characteristics (short/tall, thin/dense) and pattern application of the operator (walking towards or away from the treated area) are relevant to the dermal exposure levels of greenhouse applicators. The results showed that the highest exposure by pesticides during field applications in greenhouses, in the climatic conditions and in the crop conditions typical of a southern European country, occurs on the lower legs and front thighs of the applicators. Similar results were obtained by hierarchical cluster analysis (HCA). Drop-size seems to be very important in determining total exposure, while height and density of crops have little influence on total exposure under the conditions of the present study. No pesticide type is a major factor in total exposure. The application of multiple regression analysis (MRA) allowed assessment of the relationships between the pesticide exposure of the less affected parts of the body with the most affected parts.  相似文献   

Detection of hepatitis E virus shedding in feces of pigs at different stages of production using reverse transcription-polymerase chain reaction.     

Salceda Fernández-Barredo  Carolina Galiana  Angel García  Santiago Vega  María Teresa Gómez  María Teresa Pérez-Gracia 《Journal of veterinary diagnostic investigation》2006,18(5):462-465

The aim of this study was to determine at which production stages hepatitis E virus (HEV) is shed by the highest number of pigs and to estimate the relative risk associated with each stage. For this purpose, 146 fecal samples of pigs from 21 farms were studied. In addition, 1 sample from the manure ditch and another sample of drinking water, collected directly from the trough located in the pen, were taken from 16 farms. HEV RNA was detected in fecal samples from 34 pigs (23.29%). The production stages in which most pigs excreted HEV were weaners (41.7%) and pigs in the first month of feeding (60%). The results of the statistical analysis showed that the principal significant risk stage in HEV shedding was the first month of feeding (odds ratio [OR] 19.5, 95% CI 3.59-106.07, P = 0.001) followed by the weaners stage (OR 9.3, 95% CI .78-48.42, P = 0.008). In 8 out of 16 farms tested (50%) HEV RNA was detected in raw manure and in the water trough of only 1. Detection of HEV in manure ditches raises the concern of how to deal with manure of swine origin, because it is used as soil fertilizer.  相似文献   

Evaluation of the API 50CH and API ZYM systems for rapid characterization of Clavibacter michiganensis subsp. sepedonicus, causal agent of potato ring rot     

José Luis Palomo  María M. López  Pablo García-Benavides  Encarna Velázquez  Eustoquio Martínez-Molina 《European journal of plant pathology / European Foundation for Plant Pathology》2006,115(4):443-451

API 50CH and API ZYM systems were used to characterize fifty-three strains of Clavibacter michiganensis subsp. sepedonicus from different geographic locations and several reference strains of the same and different species, including other potato pathogens. Clavibacter michiganensis subsp. sepedonicus strains displayed a high level of homogeneity, both in carbohydrate utilization and in enzymatic activity. Using API 50CH and API ZYM it was possible to differentiate C. michiganensis subsp. sepedonicus strains from the remaining taxa analysed in this study, which included representative strains of the other subspecies of C. michiganensis as well as other bacterial pathogens affecting potatoes. Therefore, these systems could be used as an effective method to characterize C. michiganensis subsp. sepedonicus. Such a procedure would constitute an alternative system to the conventional nutritional and physiological identification tests currently included in the official methods employed in the European Union to detect and identify this bacterium. The results obtained with the API systems agreed with the current taxonomic classification of C. michiganensis, clearly separating sepedonicus from the other subspecies belonging to this species.  相似文献   

<Emphasis Type="Italic">Amaranthus leaf mottle virus</Emphasis>: 3′-end RNA sequence proves classification as distinct virus and reveals affinities within the genus <Emphasis Type="Italic">Potyvirus</Emphasis>     

Eduardo Segundo  Dietrich E. Lesemann  Germán Martín  María P. Carmona  Leticia Ruiz  Isabel M. Cuadrado  Leonardo Velasco  Dirk Janssen 《European journal of plant pathology / European Foundation for Plant Pathology》2007,117(1):81-87

Amaranthus leaf mottle virus (AmLMV) was classified as a member of the genus Potyvirus on the basis of its particle morphology, serology, and biological properties (Casetta et al., 1986). Based on these properties, an Amaranthus viridis-infecting virus isolated in Spain, causing mottle and leaf blistering as well as reduced growth has been identified as AmLMV. The 3′ terminal genomic region of this and a reference isolate from Italy has been sequenced and reveals a 95% nucleotide identity between the two isolates. The sequenced part comprises the coat protein with 281 amino acids and 315 nucleotides of the 3′ untranslated region (UTR) preceding a polyadenylated tail. Pairwise comparisons and phylogenetic analysis of the nucleotide and deduced amino acid sequences of the CP and 3′ UTR of the cloned cDNAs with those of other potyviruses shows that AmLMV is a distinct potyvirus closely related to Potato virus Y.  相似文献   

Grundflächenzuwachs und Massenzuwachs mit verschiedenen Definitionen     

Carl Mar Møller 《European Journal of Forest Research》1954,73(11-12):350-354

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Shoot development and dieback in progenies of Nothofagus obliqua     

Javier Puntieri  Javier Grosfeld  Marina Stecconi  Cecilia Brion  María Marta Azpilicueta  Leonardo Gallo  Daniel Barthélémy 《Annals of Forest Science》2007,64(8):839-844

Shoot growth and dieback were compared among progenies of nursery-grown seedlings of Nothofagus obliqua belonging to seven progenies of the same provenance (Quila-Quina, Argentina). First-year shoots consisted of one growth unit (GU) and second-year shoots of one or two GUs. The probability of development of two GU was similar for all progenies. Progenies were different in terms of shoot size, terminal bud abscission, the extent of shoot dieback after shoot extension and the node of origin of the relay shoot on the first shoot. Plants with a second-year shoot consisting of two GUs had a thicker stem and more nodes than those with single-GU shoots. The selection of N. obliqua seed trees based on architectural traits suitable for forestry development at specific sites must contemplate variability among progenies and their probabilities of successful development under different conditions.  相似文献   

Changes in understory species diversity during theNothofagus pumilio forest management cycle     

Pastur  Guillermo Martínez  Peri  Pablo Luis  Fernández  María Cecilia  Staffieri  Gabriela  Lencinas  Mará Vanessa 《Journal of Forest Research》2002,7(3):165-174

Human commercial activities must be compatible with sustainable management to achieve species conservation in the natural environment. After harvesting, diversity and abundance of understory species differ substantially from the pre-harvest situation. Have knowledge of the changes that logging produces in all components of the system allows us to develop management alternatives with reduced impacts. The objective of this work was to evaluate changes in density, biomass and diversity of understory species along a forest shelterwood cut system management cycle in aNothofagus pumilio forest. Six forest management situations (postharvesting stands and different phases of forest development) were sampled in a pureN. pumilio forest of Tierra del Fuego (Argentina) of middle-high site class (fromclass I toIII). Species number, density and biomass were modified substantially after a shelterwood cut. These were being greatest after cutting and falling substantially when the forest canopy closes during theinitial growth phase, returning to values similar to this virgin forest when themature phase was reached. The impact of the forest management on understory species diversity is analyzed and discussed. The silvicultural practices applied toN. pumilio forest in Tierra del Fuego permit the introduction of alien flora to the system, and produce great changes during the forest management cycle. To Celina Matteri, Itatí Ferraro, and Rosa Kofalt for their aid in taxonomical identification. To Juan Manuel Cellini, Ricardo Vukasovic and Gregorio Martínez Pastur for their participation in the field work; and to the sawmillLos Castores, Universidad Nacional de la Patagonia Austral, Centro Austral de Investigaciones Científicas and Instituto Nacional de Tecnología Agropecuaria for the institutional and financial support.  相似文献   

Spatial pattern of the density of Armillaria epiphytic rhizomorphs on tree collar in an oak stand     

B. Marçais  O. Caël 《Forest Pathology》2006,36(1):32-40

The density of Armillaria gallica epiphytic rhizomorphs on tree collar was mapped in a young stand of pedunculate oaks (Quercus robur). A quick method to estimate the rhizomorph density was developed, investigating 19 trees, and was used in the subsequent study. The method consists of measuring the frequency of rhizomorphs present on a grid applied to an exposed small area of the tree collar. The pattern in tree collar rhizomorph density was closely related to the pattern of stump colonization by Armillaria and stumps were therefore important as inoculum sources. Rhizomorph density on tree collar showed a strong aggregated spatial pattern which could not be explained by variability in the soil characteristics, in particular waterlogging, or by a spatial pattern in tree vigour. Moreover, rhizomorph density on tree collar did not depend on their dominance status.  相似文献