Viral properties, primary structure and phylogenetic analysis of the coat protein of an Olive latent virus 1 isolate from Olea europaea L

An Olive latent virus 1 isolate designated GM6, obtained from a Portuguese olive tree, was characterized and the coat protein gene sequenced and analysed. The purified virus particles showed to be isometric with ca. 30 nm in diameter and contained a single-stranded RNA species with ca. 3.7 kb. The dsRNA profile obtained from infected tissues showed three major species with ca. 3.7, 1.5 and 1.3 kbp. SDS-PAGE analysis revealed a major peptide with an apparent molecular mass of 32 kDa identified as the coat protein. A viral genome region containing the coat protein gene was amplified by RT-PCR and the cDNA was cloned and sequenced. The coat protein gene revealed to be 813 nucleotides long and encode a peptide with 270 amino acid residues and an estimated Mr of 29,851. Alignment of the deduced amino acid sequence with that of other necroviruses showed a higher identity with OLV-1 tulip isolate (97.7%) than with OLV-1 citrus isolate (87.7%). The consensus pattern of the coat protein 'S' domain is conserved in GM6 isolate coat protein sequence, except in amino acid 151, leucine. This is the first report on the coat protein sequence of an OLV-1 olive isolate.     

Effect of pharmacological purging on natural killer cell number and activity in human bone marrow.

We assayed natural killer (NK) cell activity and phenotype from human bone marrow (BM) following "purge" with 4-hydroperoxycyclophosphamide (4HC) at 60 micrograms/ml for 30 min in vitro. In all cases studied, lytic activity against the K562 cell line was either significantly decreased or abolished following 4HC purge. Although NK activity was significantly affected by 4HC treatment, no major differences in the phenotype between the purged and unpurged population were seen. Further, while in vitro culture of BM with IL2 resulted in a significant increment of NK activity, no IL-2 responsive cells were found in the 4HC purged BM after 14 days of culture. This study demonstrates that pharmacological purging of bone marrow results in a persistent functional decline of NK cell activity and may serve as a useful model for the study of the ontogeny of NK cells.     

IS1245 Restriction Fragment Length Polymorphism Typing of Mycobacterium avium Isolates: Proposal for Standardization

Mycobacterium avium has become a major human pathogen, primarily due to the emergence of the AIDS epidemic. Restriction fragment length polymorphism (RFLP) typing, using insertion sequence IS1245 as a probe, provides a powerful tool in the molecular epidemiology of M. avium-related infections and will facilitate well-founded studies into the sources of M. avium infections in animal and environmental reservoirs. The standardization of this technique allows computerization of IS1245 RFLP patterns for comparison on a local level and the establishment of M. avium DNA fingerprint databases for interlaboratory comparison. Moreover, by combining international DNA typing results of M. avium complex isolates from a broad spectrum of sources, long-lasting questions on the epidemiology of this major agent of mycobacterial infections will be answered.     

The serology of hepatitis C virus (HCV) infection: antibody crossreaction in the hypervariable region 1

Summary We determined the NS1/E2 N-terminal sequence including the hypervariable region 1 (HVR1) from five individuals chronically infected with HCV: two from the Czech Republic and three from Germany. From each sequence, six 12-mer overlapping peptides were synthesized and used in a peptide scan to evaluate seroreactivity of each of those patients, as well as three anti-HCV positive blood donors to the different isolates. We could show the general presence of antibodies to multiple HVR1 specific sequences reflecting the existence of multiple variants in infected persons. Finally, we observed the persistance of HCV infections in all individuals despite an active humoral response directed against the virus.     

Functional Health Literacy: Psychometric Properties of the Newest Vital Sign for Portuguese Adolescents (NVS-PTeen)

Self-management of health requires skills to obtain, process, understand, and use health-related information. Assessment of adolescents’ functional health literacy requires valid, reliable, and low-burden tools. The main objective of this study was to adapt and study the psychometric properties of the Newest Vital Sign for the Portuguese adolescents’ population (NVS-PTeen). Classic psychometric indicators of reliability and validity were combined with item response theory (IRT) analyses in a cross-sectional survey, complemented with a 3-month test-retest assessment. The NVS-PTeen was self-administered to students enrolled in grades 8 to 12 (12 to 17 years old) in a school setting. Overall, 386 students (191 girls) from 16 classes of the same school participated in the study (mean age = 14.5; SD = 1.5). Internal reliability of the NVS-PTeen was α = 0.60. The NVS-PTeen total score was positively and significantly correlated with Portuguese (r = 0.28) and mathematics scores (r = 0.31), school years (r = 0.31), and age (r = 0.19). Similar to the original scale (for the U.S.), the NVS-PTeen is composed of two dimensions, reading-related literacy and numeracy. Temporal reliability is adequate, though with a learning effect. IRT analyses revealed differences in difficulty and discriminative capacity among items, all with adequate outfit and infit values. Results showed that the NVS-PTeen is valid and reliable, sensible to inter-individual educational differences, and adequate for regular screening of functional health literacy in adolescents.     

Web Data Mining: Validity of Data from Google Earth for Food Retail Evaluation

To overcome the challenge of obtaining accurate data on community food retail, we developed an innovative tool to automatically capture food retail data from Google Earth (GE). The proposed method is relevant to non-commercial use or scholarly purposes. We aimed to test the validity of web sources data for the assessment of community food retail environment by comparison to ground-truth observations (gold standard). A secondary aim was to test whether validity differs by type of food outlet and socioeconomic status (SES). The study area included a sample of 300 census tracts stratified by SES in two of the largest cities in Brazil, Rio de Janeiro and Belo Horizonte. The GE web service was used to develop a tool for automatic acquisition of food retail data through the generation of a regular grid of points. To test its validity, this data was compared with the ground-truth data. Compared to the 856 outlets identified in 285 census tracts by the ground-truth method, the GE interface identified 731 outlets. In both cities, the GE interface scored moderate to excellent compared to the ground-truth data across all of the validity measures: sensitivity, specificity, positive predictive value, negative predictive value and accuracy (ranging from 66.3 to 100%). The validity did not differ by SES strata. Supermarkets, convenience stores and restaurants yielded better results than other store types. To our knowledge, this research is the first to investigate using GE as a tool to capture community food retail data. Our results suggest that the GE interface could be used to measure the community food environment. Validity was satisfactory for different SES areas and types of outlets.