Failure to remove bluetongue serotype 8 virus (BTV-8) from in vitro produced and in vivo derived bovine embryos and subsequent transmission of BTV-8 to recipient cows after embryo transfer

The behavior of BTV-8 in cattle is different from most other serotypes not only with regards to clinical signs but certainly with respect to virus transmission (transplacental, contact). Therefore, the possibility of virus transmission by means of embryo transfer was examined by in vitro exposure of in vitro produced and in vivo derived bovine blastocysts to BTV-8 followed by different washing protocols, including longer exposure times (up to 120 s) to 0.25% trypsin at room temperature or at 37 degrees C. None of the washing protocols used was successful in removing the viral genome completely from the in vitro produced and in vivo derived embryos as was demonstrated by real-time PCR. Moreover, BTV-8 virus was transmitted to recipient cows after embryo transfer of in vivo derived BTV8-exposed embryos, which had been subjected to routine decontamination as recommended by IETS, consisting of 5 washes in PBS followed by a double treatment of 0.25% trypsin for 45s at 37 degrees C, and an additional 5 washes in PBS with 2% FCS. This study clearly demonstrates the necessity of vigorous application of the directives for screening of potential donors and the collected embryos, especially in regions with BTV-8, to prevent transmission of the disease

Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant

Viral infections cause morbidity and mortality in allogeneic hematopoietic stem cell transplant (HSCT) recipients. We and others have successfully generated and infused T-cells specific for Epstein Barr virus (EBV), cytomegalovirus (CMV) and Adenovirus (Adv) using monocytes and EBV-transformed lymphoblastoid cell (EBV-LCL) gene-modified with an adenovirus vector as antigen presenting cells (APCs). As few as 2x105/kg trivirus-specific cytotoxic T lymphocytes (CTL) proliferated by several logs after infusion and appeared to prevent and treat even severe viral disease resistant to other available therapies. The broader implementation of this encouraging approach is limited by high production costs, complexity of manufacture and the prolonged time (4-6 weeks for EBV-LCL generation, and 4-8 weeks for CTL manufacture – total 10-14 weeks) for preparation. To overcome these limitations we have developed a new, GMP-compliant CTL production protocol. First, in place of adenovectors to stimulate T-cells we use dendritic cells (DCs) nucleofected with DNA plasmids encoding LMP2, EBNA1 and BZLF1 (EBV), Hexon and Penton (Adv), and pp65 and IE1 (CMV) as antigen-presenting cells. These APCs reactivate T cells specific for all the stimulating antigens. Second, culture of activated T-cells in the presence of IL-4 (1,000U/ml) and IL-7 (10ng/ml) increases and sustains the repertoire and frequency of specific T cells in our lines. Third, we have used a new, gas permeable culture device (G-Rex) that promotes the expansion and survival of large cell numbers after a single stimulation, thus removing the requirement for EBV-LCLs and reducing technician intervention. By implementing these changes we can now produce multispecific CTL targeting EBV, CMV, and Adv at a cost per 106 cells that is reduced by >90%, and in just 10 days rather than 10 weeks using an approach that may be extended to additional protective viral antigens. Our FDA-approved approach should be of value for prophylactic and treatment applications for high risk allogeneic HSCT recipients

Adenoviral Infections in Hematopoietic Stem Cell Transplantation

AbstractAdenoviruses are lytic DNA viruses that are ubiquitous in human communities. In total, 51 different serotypes with varying tissue tropisms have been identified. Adenovirus infections, although frequent, are rarely fatal in immunocompetent individuals who have potent innate and adaptive immunity. But in immunosuppressed individuals, adenoviruses are a significant cause of morbidity and mortality, with limited treatment options. In particular, pediatric recipients of allogeneic hematopoietic stem cell transplantation frequently develop infections early in the posttransplantation period. Because the endogenous recovery of adenovirus-specific T cells has proven important in controlling infection, we explore the potential of adoptive T-cell immunotherapy as a therapeutic strategy. We discuss the advantages and limitations of T-cell therapy for the prophylaxis and treatment of adenovirus infection posttransplantation

The use of equine chondrogenic‐induced mesenchymal stem cells as a treatment for osteoarthritis : a randomised, double‐blinded, placebo‐controlled proof‐of‐concept study

Background: There is a need to improve therapies for osteoarthritis in horses. Objectives To assess the efficacy of equine allogeneic chondrogenic-induced mesenchymal stem cells combined with equine allogeneic plasma as a novel therapy for osteoarthritis in horses. Study design: Randomised, double-blinded, placebo-controlled experiment. Methods: In 12 healthy horses, osteoarthritis was induced in the metacarpophalangeal joint using an osteochondral fragment-groove model. Five weeks after surgery, horses were randomly assigned to either an intra-articular injection with chondrogenic-induced mesenchymal stem cells + equine allogeneic plasma (= intervention) or with 0.9% saline solution (= control). From surgery until the study end, horses underwent a weekly joint and lameness assessment. Synovial fluid was collected for cytology and biomarker analysis before surgery and at Weeks 5, 5 + 1d, 7, 9 and 11. At Week 11, horses were subjected to euthanasia, and the metacarpophalangeal joints were evaluated macroscopically and histologically. Results: No serious adverse events or suspected adverse drug reactions occurred during the study. A significant improvement in visual and objective lameness was seen with the intervention compared with the control. Synovial fluid displayed a significantly higher viscosity and a significantly lower glycosaminoglycan concentration in the intervention group. Other biomarkers or cytology parameters were not significantly different between the treatment groups. Significantly less wear lines and synovial hyperaemia were present in the intervention group. The amount of cartilage oligomeric matrix protein, collagen type II and glycosaminoglycans were significantly higher in the articular cartilage of the intervention group. Main limitations: This study assessed the short-term effect of the intervention on a limited number of horses, using an osteoarthritis model. This study also included multiple statistical tests, increasing the risk of type 1 error. Conclusions: Equine allogeneic chondrogenic-induced mesenchymal stem cells combined with equine allogeneic plasma may be a promising treatment for osteoarthritis in horses

Automatic natural language generation applied to alternative and augmentative communication for online video content services using SimpleNLG for Spanish

We present our work to build the Spanish version of SimpleNLG by adapting it and creating new code to satisfy the Spanish linguistic requirements. Not only have we developed this version but also we have achieved a library that only needs the main words as input and it is able to conduct the generation process on its own. The adaptation of the library uses aLexiS, a complete and reliable lexicon with morphology that we created. On the other hand, our enhanced version uses Elsa created from the pictogram domain, which also contains syntactic and semantic information needed to conduct the generation process automatically. Both the adaptation and its enhanced version may be useful integrated in several applications as well as web applications, bringing them natural language generation functionalities. We provide a use case of the system focused on Augmentative and Alternative Communication and online video content services.Xunta de Galicia | Ref. GRC2014/046Xunta de Galicia | Ref. ED341D R2016/012Ministerio de Economía, Industria y Competitividad | Ref. TEC2016-76465-C2-2-

Prevention of pressure ulcers with a static air support surface : a systematic review

The aims of this study were to identify, assess, and summarise available evidence about the effectiveness of static air mattress overlays to prevent pressure ulcers. The primary outcome was the incidence of pressure ulcers. Secondary outcomes included costs and patient comfort. This study was a systematic review. Six electronic databases were consulted: Cochrane Library, EMBASE, PubMed (Medline), CINAHL (EBSCOhost interface), Science direct, and Web of Science. In addition, a hand search through reviews, conference proceedings, and the reference lists of the included studies was performed to identify additional studies. Potential studies were reviewed and assessed by 2 independent authors based on the title and abstract. Decisions regarding inclusion or exclusion of the studies were based on a consensus between the authors. Studies were included if the following criteria were met: reporting an original study; the outcome was the incidence of pressure ulcer categories I to IV when using a static air mattress overlay and/or in comparison with other pressure-redistribution device(s); and studies published in English, French, and Dutch. No limitation was set on study setting, design, and date of publication. The methodological quality assessment was evaluated using the Critical Appraisal Skills Program Tool. Results were reported in a descriptive way to reflect the exploratory nature of the review. The searches included 13 studies: randomised controlled trials (n = 11) and cohort studies (n = 2). The mean pressure ulcer incidence figures found in the different settings were, respectively, 7.8% pressure ulcers of categories II to IV in nursing homes, 9.06% pressure ulcers of categories I to IV in intensive care settings, and 12% pressure ulcers of categories I to IV in orthopaedic wards. Seven comparative studies reported a lower incidence in the groups of patients on a static air mattress overlay. Three studies reported a statistical (P < .1) lower incidence compared with a standard hospital mattress (10 cm thick, density 35 kg/m(3)), a foam mattress (15 cm thick), and a viscoelastic foam mattress (15 cm thick). No significant difference in incidence, purchase costs, and patient comfort was found compared with dynamic air mattresses. This review focused on the effectiveness of static air mattress overlays to prevent pressure ulcers. There are indications that these mattress overlays are more effective in preventing pressure ulcers compared with the use of a standard mattress or a pressure-reducing foam mattress in nursing homes and intensive care settings. However, interpretation of the evidence should be performed with caution due to the wide variety of methodological and/or reporting quality levels of the included studies

Engineered CAR T cell therapy for solid tumors

The adoptive transfer of T cells redirected to tumor-associated antigens via transgenic expression of chimeric antigen receptors (CARs) has produced impressive clinical responses in patients with hematologic malignances. However the successful extension of this therapy to solid tumors has proven challenging due to i) the paucity of target antigens that are tumor selective, leading to a heightened risk of “on-target, off-tumor” toxicities and, ii) the suppressive tumor microenvironment, which subverts T cell effector function. Therefore, to overcome these limitations we have programmed T cells with a combination of receptors that recognize a gene expression pattern that is unique to the tumor site and whose endodomains deliver intracellular signals 1, 2 and 3 (antigen, co-stimulation and cytokine) required for optimal T cell activation and protection from suppressive factors present at the tumor site. The current presentation will not only highlight our T cell engineering improvements but also our process optimization, including the incorporation of the G-Rex device, to facilitate the clinical and commercial development of potentially curative therapie

Susceptibility of in vitro produced hatched bovine blastocysts to infection with bluetongue virus serotype 8

Bluetongue virus serotype 8 (BTV-8), which caused an epidemic in ruminants in central Western Europe in 2006 and 2007, seems to differ from other bluetongue serotypes in that it can spread transplacentally and has been associated with an increased incidence of abortion and other reproductive problems. For these reasons, and also because BTV-8 is threatening to spread to other parts of the world, there is a need for more information on the consequences of infection during pregnancy. The aim of the present study was to investigate whether hatched (i.e. zona pellucida-free) in vitro produced bovine blastocysts at 8-9 days post insemination are susceptible to BTV-8 and whether such infection induces cell death as indicated by apoptosis. Exposure of hatched in vitro produced bovine blastocysts for 1 h to a medium containing 103.8 or 104.9 TCID50 of the virus resulted in active viral replication in between 25 and 100% of the cells at 72 h post exposure. The infected blastocysts also showed growth arrest as evidenced by lower total cell numbers and a significant level of cellular apoptosis. We conclude from this in vitro study that some of the reproductive problems that are reported when cattle herds are infected with BTV-8 may be attributed to direct infection of blastocysts and other early-stage embryos in utero