Genetic transformation of Vitis vinifera L. cvs Thompson Seedless and Chardonnay with the pear PGIP and GFP encoding genes

Transgenic plants of Vitis vinifera L. cvs. Chardonnay and Thompson Seedless expressing the β-glucuronidase gene (GUS) and either the pear polygalacturonase inhibiting protein gene (PGIP) or the green fluorescent protein gene (GFP) were produced via somatic embryogenesis. Various media and culture conditions were tested in order to develop an efficient transformation method. Best results were obtained when embryogenic callus was initiated from anthers cultured on PIV medium and maintained in PT medium. Embryogenic lines of the rootstocks Saint George, 110 Richter and Freedom and from inflorescence primordia of Chardonnay and 110 Richter were also established using the same media. Inoculation with 109 cells·ml-1 Agrobacterium resulted in a higher number of selected calli than cultures inoculated with 107 or 108 cells·ml-1. Plants were regenerated in a modified WP medium from up to 46 % of the selected callus. Approximately 80 % of the lines expressed GUS and either PGIP or GFP but a low correlation was found between β-glucuronidase and polygalacturonase inhibiting protein activities.

First report of porcine respirovirus 1 in South America.

Porcine respirovirus 1 (PRV1) is an emerging virus in pigs that has been previously described in the USA and China. There are no reports of its presence in the rest of the world. The objective of this study was to determine the occurrence of PRV1 in Chile and to determine its phylogeny. Thus, we collected samples (oral fluids, nasal swabs, and lungs) from a swine influenza A virus (IAV) surveillance program, most of which belonged to pigs with respiratory disease. The samples were analyzed by RT-PCR, and the viral sequencing was obtained using RNA whole-genome sequencing approach. Maximum likelihood phylogeny was constructed with the available references. Thirty-one of 164 samples (18.9 %) were RT-PCR positive for PRV1: 62.5 % oral fluids, 19.0 % nasal swabs, and 8.6 % lungs. All 6 farms in this study had at least one positive sample, with 6-40 % of positive results per farm, which suggests that PRV1 is disseminated in Chilean swine farms. Twenty-one of 31 (677%) PRV1-positive samples were also positive for IAV, so the role of PRV1 as secondary pathogen in respiratory disease needs to be further evaluated. Near to complete genome of two PRV1s were obtained from two farms. The phylogenies, in general, showed low bootstrap support, except the concatenated genome and the L gene trees which showed clustering of the Chilean PRV1 with Asian sequences, suggesting a close genetic relationship. This is the first report of PRV1 in the Southern Hemisphere. Further studies are necessary to determine the genetic diversity of this virus in Chile

Current collection at the shuttle orbiter during TSS‐1R high voltage charging

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/95673/1/grl10526.pd

Shuttle charging by tether controlled electron beam

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/95327/1/grl10600.pd

Association Between Physical Fitness and Bone Strength and Structure in 3- to 5-Year-Old Children

Background: The positive association between physical fitness and bone structure has been widely investigated in children and adolescents, yet no studies have evaluated this influence in young children (ie, preschoolers). Hypothesis: Fit children will present improved bone variables when compared with unfit children, and no sex-based differences will emerge in the sample. Study Design: Cross-sectional study. Level of Evidence: Level 3. Methods: Handgrip strength, standing long jump (SLJ), speed/agility, balance, and cardiorespiratory fitness (CRF) were assessed using the Assessing FITness levels in PREschoolers (PREFIT) test battery in 92 children (50 boys; age range, 3-5 years). A peripheral quantitative computed tomography scan was performed at 38% of the length of the nondominant tibia. Cluster analysis from handgrip strength, SLJ, speed/agility, and CRF was developed to identify fitness groups. Bone variables were compared between sexes and between cluster groups. The association between individual physical fitness components and different bone variables was also tested. Results: Three cluster groups emerged: fit (high values on all included physical fitness variables), strong (high strength values and low speed/agility and CRF), and unfit (low strength, speed/agility, and CRF). The fit group presented higher values than the strong and unfit groups for total and cortical bone mineral content, cortical area, and polar strength strain index (all P < 0.05). The fit group also presented a higher cortical thickness when compared with the unfit group (P < 0.05). Handgrip, SLJ, and speed/agility predicted all bone variables except for total and cortical volumetric bone mineral density. No differences were found for bone variables between sexes. Conclusion: The results suggest that global fitness in preschoolers is a key determinant for bone structure and strength but not volumetric bone mineral density. Clinical Relevance: Physical fitness is a determinant for tibial bone mineral content, structure, and strength in very young children. Performing physical fitness tests could provide useful information related to bone health in preschoolers

New GOLD classification: Longitudinal data on group assignment

Rationale: Little is known about the longitudinal changes associated with using the 2013 update of the multidimensional GOLD strategy for chronic obstructive pulmonary disease (COPD). Objective: To determine the COPD patient distribution of the new GOLD proposal and evaluate how this classification changes over one year compared with the previous GOLD staging based on spirometry only. Methods: We analyzed data from the CHAIN study, a multicenter observational Spanish cohort of COPD patients who are monitored annually. Categories were defined according to the proposed GOLD: FEV1%, mMRC dyspnea, COPD Assessment Test (CAT), Clinical COPD Questionnaire (CCQ), and exacerbations-hospitalizations. One-year follow-up information was available for all variables except CCQ data. Results: At baseline, 828 stable COPD patients were evaluated. On the basis of mMRC dyspnea versus CAT, the patients were distributed as follows: 38.2% vs. 27.2% in group A, 17.6% vs. 28.3% in group B, 15.8% vs. 12.9% in group C, and 28.4% vs. 31.6% in group D. Information was available for 526 patients at one year: 64.2% of patients remained in the same group but groups C and D show different degrees of variability. The annual progression by group was mainly associated with one-year changes in CAT scores (RR, 1.138; 95%CI: 1.074-1.206) and BODE index values (RR, 2.012; 95%CI: 1.487-2.722). Conclusions: In the new GOLD grading classification, the type of tool used to determine the level of symptoms can substantially alter the group assignment. A change in category after one year was associated with longitudinal changes in the CAT and BODE index

Envenomations by Bothrops and Crotalus Snakes Induce the Release of Mitochondrial Alarmins

Skeletal muscle necrosis is a common manifestation of viperid snakebite envenomations. Venoms from snakes of the genus Bothrops, such as that of B. asper, induce muscle tissue damage at the site of venom injection, provoking severe local pathology which often results in permanent sequelae. In contrast, the venom of the South American rattlesnake Crotalus durissus terrificus, induces a clinical picture of systemic myotoxicity, i.e., rhabdomyolysis, together with neurotoxicity. It is known that molecules released from damaged muscle might act as ‘danger’ signals. These are known as ‘alarmins’, and contribute to the inflammatory reaction by activating the innate immune system. Here we show that the venoms of B. asper and C. d. terrificus release the mitochondrial markers mtDNA (from the matrix) and cytochrome c (Cyt c) from the intermembrane space, from ex vivo mouse tibialis anterior muscles. Cyt c was released to a similar extent by the two venoms whereas B. asper venom induced the release of higher amounts of mtDNA, thus reflecting hitherto some differences in their pathological action on muscle mitochondria. At variance, injection of these venoms in mice resulted in a different time-course of mtDNA release, with B. asper venom inducing an early onset increment in plasma levels and C. d. terrificus venom provoking a delayed release. We suggest that the release of mitochondrial ‘alarmins’ might contribute to the local and systemic inflammatory events characteristic of snakebite envenomations

Protection of Spanish Ibex (Capra pyrenaica) against Bluetongue Virus Serotypes 1 and 8 in a Subclinical Experimental Infection

Many wild ruminants such as Spanish ibex (Capra pyrenaica) are susceptible to Bluetongue virus (BTV) infection, which causes disease mainly in domestic sheep and cattle. Outbreaks involving either BTV serotypes 1 (BTV-1) and 8 (BTV-8) are currently challenging Europe. Inclusion of wildlife vaccination among BTV control measures should be considered in certain species. In the present study, four out of fifteen seronegative Spanish ibexes were immunized with a single dose of inactivated vaccine against BTV-1, four against BTV-8 and seven ibexes were non vaccinated controls. Seven ibexes (four vaccinated and three controls) were inoculated with each BTV serotype. Antibody and IFN-gamma responses were evaluated until 28 days after inoculation (dpi). The vaccinated ibexes showed significant (P<0.05) neutralizing antibody levels after vaccination compared to non vaccinated ibexes. The non vaccinated ibexes remained seronegative until challenge and showed neutralizing antibodies from 7 dpi. BTV RNA was detected in the blood of non vaccinated ibexes from 2 to the end of the study (28 dpi) and in target tissue samples obtained at necropsy (8 and 28 dpi). BTV-1 was successfully isolated on cell culture from blood and target tissues of non vaccinated ibexes. Clinical signs were unapparent and no gross lesions were found at necropsy. Our results show for the first time that Spanish ibex is susceptible and asymptomatic to BTV infection and also that a single dose of vaccine prevents viraemia against BTV-1 and BTV-8 replication

Genomic Diversity of Mycobacterium tuberculosis Complex Strains in Cantabria (Spain), a Moderate TB Incidence Setting

Background Tuberculosis (TB) control strategies are focused mainly on prevention, early diagnosis, compliance to treatment and contact tracing. The objectives of this study were to explore the frequency and risk factors of recent transmission of clinical isolates of Mycobacterium tuberculosis complex (MTBC) in Cantabria in Northern Spain from 2012 through 2013 and to analyze their clonal complexity for better understanding of the transmission dynamics in a moderate TB incidence setting. Methods DNA from 85 out of 87 isolates from bacteriologically confirmed cases of MTBC infection were extracted directly from frozen stocks and genotyped using the mycobacterial interspersed repetitive units-variable number tandem repeat (MIRU-VNTR) method. The MIRUVNTRplus database tool was used to identify clusters and lineages and to build a neighbor joining (NJ) phylogenetic tree. In addition, data were compared to the SITVIT2 database at the Pasteur Institute of Guadeloupe. Results The rate of recent transmission was calculated to 24%. Clustering was associated with being Spanish-born. A high prevalence of isolates of the Euro-American lineage was found. In addition, MIRU-VNTR profiles of the studied isolates corresponded to previously found MIRU-VNTR types in other countries, including Spain, Belgium, Great Britain, USA, Croatia, South Africa and The Netherlands. Six of the strains analyzed represented clonal variants. Conclusion Transmission of MTBC is well controlled in Cantabria. The majority of TB patients were born in Spain. The population structure of MTBC in Cantabria has a low diversity of major clonal lineages with the Euro-American lineage predominating