Differentiation of equine bone marrow derived mesenchymal stem cells increases the expression of immunogenic genes

Mesenchymal stem cells (MSCs) are a promising treatment for equine musculoskeletal injuries because of their ability to regulate the inflammation and to differentiate into other cell types. Since interest in allogeneic therapy is rising, concerns about MSC immunogenicity need to be addressed. Differentiated MSCs from several species increase their expression of immunogenic molecules and induce alloresponses, but equine MSC immunogenic profile after differentiation has not been reported. Therefore, the aim of this study was to assess the gene expression of immunogenic markers in tri-lineage differentiated equine bone marrow derived MSCs (eBM-MSCs). For this purpose, eBM-MSCs (n = 4) were differentiated into osteoblasts, adipocytes and chondrocytes. Differentiation was confirmed by specific staining and gene expression of lineage-related markers. Subsequently, gene expression of MHC-I, MHC-II, CD40 and CD80 was analyzed in undifferentiated (control) and tri-lineage differentiated eBM-MSCs. Osteogenesis and adipogenesis, but not chondrogenesis, significantly upregulated MHC-I; MHC-II expression significantly increased in the three lineages, while CD40 and CD80 expression did not change. Despite this, MHC-I and MHC-II upregulation after differentiation might lead to increased immunogenicity and risk of allorecognition, either eBM-MSCs differentiate in vivo after administration or they are differentiated prior to administration, with potential negative consequences for effectiveness and safety of allogeneic therapy

The amino terminal end determines the stability and assembling capacity of eukaryotic ribosomal stalk proteins P1 and P2

The eukaryotic ribosomal proteins P1 and P2 bind to protein P0 through their N-terminal domain to form the essential ribosomal stalk. A mutational analysis points to amino acids at positions 2 and 3 as determinants for the drastic difference of Saccharomyces cerevisiae P1 and P2 half-life, and suggest different degradation mechanisms for each protein type. Moreover, the capacity to form P1/P2 heterodimers is drastically affected by mutations in the P2β four initial amino acids, while these mutations have no effect on P1β. Binding of P2β and, to a lesser extent, P1β to the ribosome is also seriously affected showing the high relevance of the amino acids in the first turn of the NTD α-helix 1 for the stalk assembly. The negative effect of some mutations on ribosome binding can be reversed by the presence of the second P1/P2 couple in the ribosome, indicating a stabilizing structural influence between the two heterodimers. Unexpectedly, some mutations totally abolish heterodimer formation but allow significant ribosome binding and, therefore, a previous P1 and P2 association seems not to be an absolute requirement for stalk assembly. Homology modeling of the protein complexes suggests that the mutated residues can affect the overall protein conformation

In vivo assembling of bacterial ribosomal protein L11 into yeast ribosomes makes the particles sensitive to the prokaryotic specific antibiotic thiostrepton

Article available at http://dx.doi.org/10.1093/nar/gkm773Eukaryotic ribosomal stalk protein L12 and its bacterial orthologue L11 play a central role on ribosomal conformational changes during translocation. Deletion of the two genes encoding L12 in Saccharomyces cerevisiae resulted in a very slow-growth phenotype. Gene RPL12B, but not the RPL12A, cloned in centromeric plasmids fully restored control protein level and the growth rate when expressed in a L12-deprived strain. The same strain has been transformed to express Escherichia coli protein EcL11 under the control of yeast RPL12B promoter. The bacterial protein has been found in similar amounts in washed ribosomes from the transformed yeast strain and from control E. coli cells, however, EcL11 was unable to restore the defective acidic protein stalk composition caused by the absence of ScL12 in the yeast ribosome. Protein EcL11 induced a 10% increase in L12-defective cell growth rate, although the in vitro polymerizing capacity of the EcL11-containing ribosomes is restored in a higher proportion, and, moreover, the particles became partially sensitive to the prokaryotic specific antibiotic thiostrepton. Molecular dynamic simulations using modelled complexes support the correct assembly of bacterial L11 into the yeast ribosome and confirm its direct implication of its CTD in the binding of thiostrepton to ribosomesThis work was funded by Ministerio de Educación y Ciencia, Spain (BFU2006-00365 to J.P.G.B., GEN2003-206420-C09-08 and BIO2005-0576 to A.R.O.); Fundación Ramón Areces (institutional grant to CBMSO)Peer reviewe

Analysis of different ventilation strategies and CO2 distribution in a naturally ventilated classroom

CO2 monitoring has proven to be an effective and affordable way of controlling air ventilation rates, a paramount task for minimizing airborne contagions in indoor shared spaces. In this work, the CO2 distribution in a naturally-ventilated classroom has been thoroughly characterized, gaining information not only on the effectiveness of diverse ventilation strategies but also on the expected differences between CO2 values when varying the sampling location within the room. The results confirm that an adequate renewal of the air in the room requires the use of cross-ventilation, with openings in different walls. Furthermore, it was found that ventilation is optimized, for a given total opening area, when the openings are distributed as much as possible among different windows. For most of the studied conditions, a global windows opening area of 1.24 m2 with an open door was typically enough to yield CO2 concentrations below 700 ppm. The CO2 readings displayed a noticeable and consistent dependency on the sampling height, with below-average values at 0.75 m, the highest concentrations at 1.5 m, and levels close to the average when sampling at a height of 2.2 m. For a given height, the influence of the sampling location within the room was weaker, and more dependent on the specific ventilation strategy applied. However, the tests consistently showed CO2 records significantly lower for sensors installed on the walls. Besides a detailed spatial and temporal characterization of the ventilation process under different ventilation strategies, these results are thought to provide useful and novel information for a judicious placement of CO2 monitoring systems

SARS-CoV-2 congenital infection and pre-eclampsia-like syndrome in dichorionic twins: A case report and review of the literature

Although the route of transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is mainly respiratory, vertical transmission seems possible.1 We report the case of a woman with a dichorionic diamniotic twin pregnancy admitted to Hospital Clínico Universitario Lozano Blesa at 38+4 weeks of gestation due to severe pre-eclampsia in the context of a SARS-CoV-2 infection (positive nasopharyngeal PCR; Viasure, CerTest Biotec., Zaragoza, Spain) with a probable transplacental transmission of the virus to both twins..

Simultaneous optimization of circadian and color performance for smart lighting systems design

We present in this work a method to design lighting sources that can be adapted to different temperatures of color and, simultaneously, with a tunable circadian character. We obtained an acceptable range of tuning in both parameters compared to the bibliography. This kind of lighting source has potential applications particularly in building lighting, but also in farming or agriculture. At the same time, we have shown the possibilities of multiobjective optimizations in the lighting industry. The optimization has been developed using the Genetic Algorithm and multiobjective merit functions. The lighting source is able to work under two different regimes regarding the circadian effect, with a design based on a combination of two monochromatic and two white Lighting Emitting Diodes (enough for controlling the circadian character and the color performance at the same time). A prototype, which can be manually or automatically controlled, has been also implemented and evaluated, with a performance in terms of color coordinates very close to the daylight, showing a modulation of the Circadian Efficacy of Radiation between 6% and 16%, and a Color Rendering Index above 80%

Fast continuous alcohol amination employing a hydrogen borrowing protocol

A continuous flow method for the direct conversion of alcohols to amines via a hydrogen borrowing approach is reported.The authors are grateful to Eli Lilly & Co. (RL) and the EPSRC (SVL, grants EP/K009494/1, EP/M004120/1 and EP/K039520/1) for financial support. This work was supported by Eli Lilly & Co. through the Lilly Research Award Program (LRAP)

Inflammation affects the viability and plasticity of equine mesenchymal stem cells: Possible implications in intra-articular treatments

Mesenchymal stem cells (MSCs) are gaining relevance for treating equine joint injuries because of their ability to limit inflammation and stimulate regeneration. Because inflammation activates MSC immunoregulatory function, proinflammatory priming could improve MSC efficacy. However, inflammatory molecules present in synovial fluid or added to the culture medium might have deleterious effects on MSCs. Therefore, this study was conducted to investigate the effects of inflammatory synovial fluid and proinflammatory cytokines priming on viability and plasticity of equine MSCs. Equine bone marrow derived MSCs (eBM-MSCs) from three animals were cultured for 72 h in media supplemented with: 20% inflammatory synovial fluid (SF); 50 ng/mL IFN-¿ and TNF-a (CK50); and 20 ng/mL IFN-¿ and TNF-a (CK20). Proliferation assay and expression of proliferation and apoptosis-related genes showed that SF exposed-eBM-MSCs maintained their viability, whereas the viability of CK primed-eBM-MSCs was significantly impaired. Tri-lineage differentiation assay revealed that exposure to inflammatory synovial fluid did not alter eBM-MSCs differentiation potential; however, eBM-MSCs primed with cytokines did not display osteogenic, adipogenic or chondrogenic phenotype. The inflammatory synovial environment is well tolerated by eBM-MSCs, whereas cytokine priming negatively affects the viability and differentiation abilities of eBM-MSCs, which might limit their in vivo efficacy

Comparison of autologous bone marrow and adipose tissue derived mesenchymal stem cells, and platelet rich plasma, for treating surgically induced lesions of the equine superficial digital flexor tendon

Several therapies have been investigated for equine tendinopathies, but satisfactory long term results have not been achieved consistently and a better understanding of the healing mechanism elicited by regenerative therapies is needed. The aim of this study was to assess the separate effects of autologous bone marrow (BM) and adipose tissue (AT) derived mesenchymal stem cells (MSCs), and platelet rich plasma (PRP), for treating lesions induced in the superficial digital flexor tendon (SDFT) of horses. Lesions were created surgically in both SDFTs of the forelimbs of 12 horses and were treated with BM-MSCs (six tendons), AT-MSCs (six tendons) or PRP (six tendons). The remaining six tendons received lactated Ringer''s solution as control. Serial ultrasound assessment was performed prior to treatment and at 2, 6, 10, 20 and 45 weeks post-treatment. At 45 weeks, histopathology and gene expression analyses were performed. At week 6, the ultrasound echogenicity score in tendons treated with BM-MSCs suggested earlier improvement, whilst all treatment groups reached the same level at week 10, which was superior to the control group. Collagen orientation scores on histological examination suggested a better outcome in treated tendons. Gene expression was indicative of better tissue regeneration after all treatments, especially for BM-MSCs, as suggested by upregulation of collagen type I, decorin, tenascin and matrix metalloproteinase III mRNA. Considering all findings, a clear beneficial effect was elicited by all treatments compared with the control group. Although differences between treatments were relatively small, BM-MSCs resulted in a better outcome than PRP and AT-MSCs