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

Measurement method of optical properties of ex vivo biological tissues of rats in the near-infrared range

An optical fiber-based supercontinuum setup and a custom-made spectrophotometer that can measure spectra from 1100 to 2300 nm, are used to describe attenuation properties from different ex vivo rat tissues. Our method is able to differentiate between scattering and absorption coefficients in biological tissues. Theoretical assumptions combined with experimental measurements demonstrate that, in this infrared range, tissue attenuation and absorption can be accurately measured, and scattering can be described as the difference between both magnitudes. Attenuation, absorption, and scattering spectral coefficients of heart, brain, spleen, retina, and kidney are given by applying these theoretical and experimental methods. Light through these tissues is affected by high scattering, resulting in multiple absorption events, and longer wavelengths should be used to obtain lower attenuation values. It can be observed that the absorption coefficient has a similar behavior in the samples under study, with two main zones of absorption due to the water absorption bands at 1450 and 1950 nm, and with different absolute absorption values depending on the constituents of each tissue. The scattering coefficient can be determined, showing slight differences between retina and brain samples, and among heart, spleen and kidney tissues

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

¿Se deberían replantear las recomendaciones relativas al uso sedentario del tiempo de pantalla en adolescentes?

El objetivo fue analizar el tiempo medio de uso diario de cuatro medios tecnológicos de pantalla (i.e., TV, ordenador, videojuegos y móvil) e identificar el grado de cumplimiento de las recomendaciones de pantalla, tanto de manera independiente como en su conjunto. Asimismo, se analizaronlas diferencias en función del tipo de jornada y el género. Participaron 105 estudiantes (M=13.05±0.59)que cumplimentaron un cuestionario referente al tiempo de pantalla de dichos medios tecnológicos. Los adolescentes de esta muestra pasan untiempo medio de uso diario de 407 minutos de medios tecnológicos de pantalla, siendo la TV el medio más utilizado.Ninguno de los adolescentes cumple las recomendaciones de tiempo de pantalla. Asimismo, existe un mayor de tiempo de uso tecnológico los fines de semana y por parte de los chicos. Se discute la necesidad de actualización de las recomendaciones relativas al uso del tiempo de pantalla en adolescentes. The aim of this study was to analyze the daily sedentary screen time of four electronic devices behaviors (i.e., TV, computer, video games, and mobile), and the proportion of adolescents who meet recommendations for screen time either independently or combined. Differences according to the type of day and gender were analyzed. One hundred five students (M=13.05 +/- 0.59) completed a self-reported questionnaire about these sedentary screen time behaviors. Adolescents reported an average daily rime of 407 minutes of sedentary screen time behaviors, with TV being the most used electronic device. Results showed that all adolescents failed to meet screen time recommendations. Greater values of sedentary screen time behaviors were reported on weekends days and in boys than in weekdays and in girls respectively. The need to update the sedentary screen time guidelines in adolescents is discussed

Single-shot d-scan technique for ultrashort laser pulse characterization using transverse second-harmonic generation in random nonlinear crystals

We demonstrate a novel dispersion-scan (d-scan) scheme for single-shot temporal characterization of ultrashort laser pulses. The novelty of this method relies on the use of a highly dispersive crystal featuring antiparallel nonlinear domains with a random distribution and size. This crystal, capable of generating a transverse second-harmonic signal, acts simultaneously as the dispersive element and the nonlinear medium of the d-scan device. The resulting in-line architecture makes the technique very simple and robust, allowing the acquisition of single-shot d-scan traces in real time. The retrieved pulses are in very good agreement with independent frequency-resolved optical grating measurements. We also apply the new single-shot d-scan to a terawatt-class laser equipped with a programmable pulse shaper, obtaining an excellent agreement between the applied and the d-scan retrieved dispersions

¿Se deberían replantear las recomendaciones relativas al uso sedentario del tiempo de pantalla en adolescentes?

The aim of thisstudy was to analyze the daily sedentary screen time of four electronicdevices behaviors (i.e., TV, computer, videogames, and mobile),and the proportion of adolescents who meet recommendations for screen time either independently or combined. Differences according to the type of day and gender were analyzed. One hundred fivestudents (M=13.05±0.59) completed a self-reported questionnaire about these sedentary screen time behaviors. Adolescents reported an average daily time of 407 minutes of sedentary screen time behaviors, with TV being the most used electronic device. Results showed that all adolescents failed to meet screen time recommendations. Greater values of sedentary screen time behaviors were reported on weekends days and in boys than in weekdays and in girls respectively. The need to update the sedentary screen time guidelines in adolescents is discussed.El objetivo fue analizar el tiempo medio de uso diario de cuatro medios tecnológicos de pantalla (i.e., TV, ordenador, videojuegos y móvil) e identificar el grado de cumplimiento de las recomendaciones de pantalla, tanto de manera independiente como en su conjunto. Asimismo, se analizaronlas diferencias en función del tipo de jornada y el género. Participaron 105 estudiantes (M=13.05±0.59)que cumplimentaron un cuestionario referente al tiempo de pantalla de dichos medios tecnológicos. Los adolescentes de esta muestra pasan untiempo medio de uso diario de 407 minutos de medios tecnológicos de pantalla, siendo la TV el medio más utilizado.Ninguno de los adolescentes cumple las recomendaciones de tiempo de pantalla. Asimismo, existe un mayor de tiempo de uso tecnológico los fines de semana y por parte de los chicos. Se discute la necesidad de actualización de las recomendaciones relativas al uso del tiempo de pantalla en adolescentes

El gen tpi como herramienta en los estudios epidemiolo´gicos de la giardiosis

Giardia duodenalis es un protozoo que causa infección en humanos y animales, que se puede transmitir por vía hídrica, de persona a persona o por contacto con animales, siendo una de las infecciones intestinales más frecuentes en nuestro país, por lo que supone una preocupación de Salud Pública. Su estudio epidemiológico, requiere la caracterización molecular de los parásitos, utilizando genes con gran variabilidad como el que codifica la triosafosfatoisomerasa (tpi) y analizando la homología entre aislamientos. El objetivo del trabajo es establecer el criterio de identidad que permita la comparación epidemiológica de los aislamientos de Giardia. Se recogieron 2-3 muestras de heces en días alternos, de 26 pacientes con giardiosis. Tras la extracción de ADN, se amplificaron por técnicas de PCR, un fragmento del gen tpi y un fragmento del gen de la beta-giardina (bg), que se utilizó como comparación. Los fragmentos obtenidos fueron secuenciados y las secuencias analizadas con los programas BioEdit y DnaSP v.5.0. Las secuencias del gen tpi mostraron una elevada divergencia, con valores de diversidad ¿ entre 0 y 0, 21219. La aparición de picos múltiples en el cromatograma, indicaron la presencia de varios clones en la misma muestra. Las diferencias entre aislamientos del mismo paciente fueron iguales o mayores que las encontradas para el conjunto de todas las muestras. La variabilidad del gen tpi no permite establecer unos criterios de identidad, necesarios para la identificación de aislamientos. Las infecciones mixtas intragenotipo ocurren de una forma muy frecuente, sugiriendo una implicación de la vía ambiental como principal fuente de transmisión o una variación genética muy elevada. Giardia duodenalis is a protozoon that causes infection in humans and animals. It can be transmitted by contaminated water, from person to person or by contact with animals; it being the cause one of the most common intestinal infections in our country, so it is a public health concern. The epidemiological study thereof requires the molecular characterization of parasites, using genes with great variability, such as the one that codes triosephosphate isomerase (tpi), and analizing the homology between isolates. The purpose of this work is to establish the identity criterion for epidemiological comparison of Giardia isolates. 2-3 stool samples were collected in alternate days from 26 patients with giardiasis. After DNA extraction, a fragment of the tpi gene and a fragment of the beta-giardin (bg) gene-used for comparison purposes-were amplified by means of PCR techniques. The obtained fragments were sequenced and the sequences analyzed with the BioEdit and DnaSP v.5.0 software. The tpi gene sequences showed a high divergence, with values of diversity ¿ ranging from 0 to 0.21219. The appearance of multiple peaks in the chromatogram points to the presence of various clones in the same sample. The differences between isolates from the same patient where equal or higher than those found for the collection of all samples. The variability of the tpi gene does not allow identity criteria to be established, which are necessary for isolate identification. Mixed intragenotype infections occur very frequently, which suggests the environmental path is the principal path of transmission and/or there is very high genetic variability

Ongoing outbreak of hepatitis A in Italy: preliminary report as of 31 May 2013

Since January 2013, an unusual increase in hepatitis A cases has been detected in northern Italy. A total number of 352 cases were reported to the integrated surveillance system between January and the end of May 2013 and this represents a 70% increase compared to the same period of the previous year. The outbreak is ongoing and the public health authorities are continuing their investigations to establish the transmission vehicle and to control the outbreak