Detection, quantification and genetic variability of Mycoplasma hyopneumoniae from apparently healthy and pneumonic swine

Mycoplasma hyopneumoniae is the causative agent of the Porcine Enzootic Pneumonia. However, this mycoplasma can be detected in healthy and symptomatic pigs, that difficults the conclusion for the etiology of this disease. In the present study we aimed to detect, quantify and do molecular analyses of M. hyopneumoniae strains in respiratory clinical samples recovered from healthy pigs and from those with pneumonia or other respiratory symptoms. The analytical sensitivity and specificity of PCR assays directed to Mollicutes detection and porcine mycoplasmas identification in clinical samples were evaluated. The identification of M. hyopneumoniae in the samples was performed using different molecular approaches, Multiplex PCR, Real Time PCR and Multilocus Variable-Number Tandem-Repeat amplification. Molecular characterization of the strains was achieved by determining and comparing the VNTR copy number directly in the samples. The highest number of samples positive to M. hyopneumoniae was identified by the multilocus VNTR amplification assay using labeled primers, followed by capillary electrophoresis. The highest concentration of M. hyopneumoniae was detected in pneumonic lungs (2, 3 * 108 genome copies /mL). The VNTR copy number analysis demonstrated that despite the high genetic variability of the M. hyopneumoniae strains, predominant strains in the swine farms could be identified by means of the VNTR copy number analysis of P97R1 and P146R3. (English)Molecular differences among Mycoplasma hyopneumoniae strains present in pneumonic lungs of swine have been largely studied. However, no comparative studies concerning the strains present in apparently healthy pigs have been carried out. This study aimed to detect, quantify and perform molecular analysis of M. hyopneumoniae strains in pig lungs with and without pneumonic lesions. The detection of M. hyopneumoniae was performed using multiplex PCR (YAMAGUTI, 2008), real-time PCR (STRAIT et al., 2008) and multiple VNTR amplification (VRANCKX et al., 2011). Molecular characterization of the strains was achieved by analysis of the VNTR copy number in P97R1, P146R3, H2R1 and H4. M. hyopneumoniae was detected in samples from healthy and pneumonic pigs and the amount of M. hyopneumoniae positive samples detected varied with the type of assay. The greater number of positive samples was identified by the multiple VNTR amplification combined with capillary electrophoresis. Using real-time PCR, 4.9*104 M. hyopneumoniae genome copies/mL was detected in apparently healthy lungs. A mean quantity of 3.9*106 M. hyopneumoniae genome copies/mL was detected in pneumonic lungs. The analysis of VNTR copy number demonstrated a high genetic variability of the M. hyopneumoniae strains present in apparently healthy and pneumonic lungs. Strains having 3 VNTR copy number in P97R1, were detected only in pneumonic lungs and strains having 40 and 43 VNTR copy number in P146R3 were detected only in apparently healthy lungs. Despite the genetic variability of M. hyopneumoniae, predominant strains in the swine farms could be identified.As diferenças moleculares entre as estirpes de Mycoplasma hyopneumoniae presentes em pulmões de suínos com pneumonia tem sido estudadas. Porém, estudos comparativos relativos as estirpes presentes nos suínos aparentemente saudáveis não foram levados a cabo. O objetivo do estudo foi a detecção, quantificação e analise molecular de M. hyopneumoniae nos pulmões suínos com e sem lesões pneumônicas. Para a detecção de M. hyopneumoniae usaramse o PCR Multiplo (YAMAGUTI, 2008), o PCR a Tempo Real (STRAIT et al., 2008) e a amplificação de múltiplo VNTR (VRANCKX et al., 2011). A caracterização molecular das estirpes foi realizada mediante a análise do número de copias de VNTR em P97R1, P146R3, H2R1 e H4. O M. hyopneumoniae foi detectado em amostras de suínos saudáveis e pneumônicos e a quantidade de M. hyopneumoniae nas amostras positivas variou com o tipo de ensaio. O maior número de amostras positivas foi identificado pela amplificação de múltiplas VNTR combinado com a eletroforese de capilares. Usando o PCR a Tempo Real, 4.9*104 copias de genoma/mL de M. hyopneumoniae foram detectadas em pulmões aparentemente saudáveis. Uma quantidade média de 3.9*106 copias de genoma/mL de M. hyopneumoniae foi detectada em pulmões pneumônicos. A análise do número de copias de VNTR demonstrou uma elevada variabilidade

The Incorporation of etanercept into a porous tri-layer scaffold for restoring and repairing cartilage tissue

Producción CientíficaCartilage diseases currently affect a high percentage of the world’s population. Almost all of these diseases, such as osteoarthritis (OA), cause inflammation of this soft tissue. However, this could be controlled with biomaterials that act as an anti-inflammatory delivery system, capable of dosing these drugs over time in a specific area. The objective of this study was to incorporate etanercept (ETA) into porous three-layer scaffolds to decrease the inflammatory process in this soft tissue. ETA is a blocker of pro-inflammatory cytokines, such as tumour necrosis factor alpha (TNF-α) and interleukin 6 (IL-6). For this reason, the scaffold was built based on natural polymers, including chitosan and type I collagen. The scaffold was grafted next to subchondral bone using hydroxyapatite as filler. One of the biomaterials obtained was also crosslinked to compare its mechanical properties with the non-treated one. Both samples’ physicochemical properties were studied with SEM, microCT and photoacoustic imaging, and their rheological properties were also compared. The cell viability and proliferation of the human chondrocyte C28/I2 cell line were studied in vitro. An in vitro and in vivo controlled release study was evaluated in both specimens. The ETA anti-inflammatory effect was also studied by in vitro TNF-α and IL-6 production. The crosslinked and non-treated scaffolds had rheological properties suitable for this application. They were non-cytotoxic and favoured the in vitro growth of chondrocytes. The in vitro and in vivo ETA release showed desirable results for a drug delivery system. The TNF-α and IL-6 production assay showed that this drug was effective as an anti-inflammatory agent. In an in vivo OA mice model, safranin-O and fast green staining was carried out. The OA cartilage tissue improved when the scaffold with ETA was grafted in the damaged area. These results demonstrate that this type of biomaterial has high potential for clinical applications in tissue engineering and as a controlled drug delivery system in OA articular cartilage.European Union through the Erasmus PLUS doctoral fellowship (project 2015-1-NL01-KA 107-008639)VIDI personal grant (project 723.012.110

Detection, quantification and genetic variability of Mycoplasma hyopneumoniae from apparently healthy and pneumonic swine

Molecular differences among Mycoplasma hyopneumoniae strains present in pneumonic lungs of swine have been largely studied. However, no comparative studies concerning the strains present in apparently healthy pigs have been carried out. This study aimed to detect, quantify and perform molecular analysis of M. hyopneumoniae strains in pig lungs with and without pneumonic lesions. The detection of M. hyopneumoniae was performed using multiplex PCR (YAMAGUTI, 2008), real-time PCR (STRAIT et al., 2008) and multiple VNTR amplification (VRANCKX et al., 2011). Molecular characterization of the strains was achieved by analysis of the VNTR copy number in P97R1, P146R3, H2R1 and H4. M. hyopneumoniae was detected in samples from healthy and pneumonic pigs and the amount of M. hyopneumoniae positive samples detected varied with the type of assay. The greater number of positive samples was identified by the multiple VNTR amplification combined with capillary electrophoresis. Using real-time PCR, 4.9*104 M. hyopneumoniae genome copies/mL was detected in apparently healthy lungs. A mean quantity of 3.9*106 M. hyopneumoniae genome copies/mL was detected in pneumonic lungs. The analysis of VNTR copy number demonstrated a high genetic variability of the M. hyopneumoniae strains present in apparently healthy and pneumonic lungs. Strains having 3 VNTR copy number in P97R1, were detected only in pneumonic lungs and strains having 40 and 43 VNTR copy number in P146R3 were detected only in apparently healthy lungs. Despite the genetic variability of M. hyopneumoniae, predominant strains in the swine farms could be identified

Modificación de conocimientos sobre salud bucal en adolescentes de la Secundaria Básica "Orlando Fernández Badell"

Se realizó una intervención educativa en 40 estudiantes de la Secundaria Básica "Orlando Fernández Badell" de Santiago de Cuba -- seleccionados por muestro aleatorio simple --, de enero a julio del 2014, a fin de modificar sus conocimientos sobre salud bucal. En los adolescentes se efectuó un examen bucal y se aplicó un cuestionario, antes y después de presentar el programa de actividades educativas, que garantizó la motivación y el incremento significativo de los conocimientos sobre los aspectos abordados (95,0 %). En el estudio se obtuvo que la mayoría de los jóvenes desconocían información necesaria acerca de la higiene bucal y la dieta cariogénica; de modo que la actividad educativa desarrollada fue oportuna y efectiva, pues aportó excelentes resultados en la transformación de los conocimientos sobre salud buca

Algunas consideraciones sobre el surgimiento y la evolución de la medicina natural y tradicional

La medicina natural, o naturismo, es un método curativo que utiliza múltiples medios naturales para permitir que el individuo adquiera un nivel máximo de salud. El sistema naturalista para curar enfermedades se basa en el uso de los recursos del medio ambiente para regular la dieta, la respiración, el ejercicio y los baños, y además saber emplear las fuerzas que eliminan todos los productos nocivos que se acumulan en el organismo, de manera que sea recuperada la vitalidad y se alcance un nivel de salud adecuado. A través de este trabajo se puede constatar que a medida que el hombre ha ido evolucionando, se han desarrollado las diferentes ramas de la medicina natural y tradiciona

The Effects of crosslinking on the rheology and cellular behavior of polymer-based 3D-multilayered scaffolds for restoring articular cartilage

Producción CientíficaPolymer-based tri-layered (bone, intermediate and top layers) scaffolds used for the restoration of articular cartilage were prepared and characterized in this study to emulate the concentration gradient of cartilage. The scaffolds were physically or chemically crosslinked. In order to obtain adequate scaffolds for the intended application, the impact of the type of calcium phosphate used in the bone layer, the polymer used in the intermediate layer and the interlayer crosslinking process were analyzed. The correlation among SEM micrographs, physical-chemical characterization, swelling behavior, rheological measurements and cell studies were examined. Storage moduli at 1 Hz were 0.3–1.7 kPa for physically crosslinked scaffolds, and 4–5 kPa (EDC/NHS system) and 15–20 kPa (glutaraldehyde) for chemically crosslinked scaffolds. Intrinsic viscoelasticity and poroelasticity were considered in discussing the physical mechanism dominating in different time/frequency scales. Cell evaluation showed that all samples are available as alternatives to repair and/or substitute cartilage in articular osteoarthritis.The European Union through the Erasmus MUNDUS LINDO post-doctoral fellowship,(project code ML14PD1157) and Erasmus PLUS doctoral fellowship, (project code 2015-1-NL01-KA 107-008639)European Union’s Horizon 2020 research and innovation program. grant agreement - H2020- MSCA-RISE (644373—PRISAR), H2020-MSCA-RISE (777682—CANCER), H2020-WIDESPREAD-05- 2017-Twinning (807281—ACORN), H2020-WIDESPREAD-2018-03 (852985—SIMICA), H2020-SCA RISE-2016 (734684—CHARMED) and MSCA-ITN-2015-ETN (675743—ISPIC), 861190 (PAVE), 857894 (CAST), 859908 (NOVA-MRI); 872860 (PRISAR2)VIDI grant (project number 723.012.110

A novel information criterion to elucidate a drug delivery mechanism from poly (acrylamide-co-2-hydroxyethyl methacrylate) reinforced with hydroxyapatite composite

Producción CientíficaHydrophilic composites of poly (acrylamide-co-2-hydroxyethyl methacrylate) reinforced with hydroxyapatite were synthesized, characterized and evaluated as drug delivery systems. The results show a strong dependence of the polymer composition, the degree of crosslinking and the homogeneity of the particles dispersion (filler or the mixed drug) in the characteristics that were analyzed. The mechanical properties were better while the swelling and the diffusion coefficient were worse than hydrogels with cefazolin of similar compositions. Additionally, a new information criterion was used to elucidate the best fit. The drug diffusion into the medium was anomalous with a tendency towards Case II transport, indicating that under certain conditions zero-order diffusion could be achieved. This fact suggests that these materials could be used in the manufacture of medical devices for the controlled delivery of drugs and active or biological principles, significantly improving therapeutic procedures today.2020-12-062020-12-06European Union (Erasmus PLUS project NL01-KA 107-008639; Erasmus MUNDUS LINDO ML14PD1157

Cuadernos del CIPS 2008 : experiencias de investigación social en Cuba

A XXV años de su fundación, el Centro de Investigaciones Psicológicas y Sociológicas (CIPS)se ha propuesto hacer un alto en el camino para valorar y socializar el trabajo realizado en esto años de estudio de la sociedad cubana. En Los Cuadernos del CIPS 2008: Experiencias de investigación social en Cuba, se profundiza en el abordaje histórico de categorías y problemáticas vitales de la sociedad cubana como la estructura social y las desigualdades, la familia, la religión, la subjetividad social, la juventud, el trabajo con las organizaciones, el desarrollo local y comunitario, así como la creatividad y el desarrollo humano. La obra ofrece además, una información pormenorizada sobre las investigaciones realizadas, las metodologías producidas y los reconocimientos recibidos en este cuarto de siglo. Incluye tres balances críticos sobre generalidades de nuestro quehacer, que abundan en el análisis de las principales líneas de trabajo, la inserción en el contexto de las ciencias sociales internacionales y en los retos futuros

El derecho a los derechos : infancias y adolescencias en Cuba

«El derecho a los derechos. Infancias y adolescencias en Cuba», constituye un título atractivo y seductor, en el que se ofrece al lector una mirada crítica a retos, problemáticas y cuestiones teórico-prácticas en la protección integral de los niños, niñas y adolescentes. Profesionalidad, originalidad, impronta y opinión personal de cada uno de los autores signaron la concepción del libro. De igual forma, un lenguaje apropiado que se sustrae de tecnicismos barrocos y excesivos para facilitar la lectura. Es una obra pensada desde la academia para impactar la realidad social. (...) engrosa los estudios sobre infancia y adolescencia existentes, es expresión de un saber especializado, tanto a nivel de la investigación científica como de la práctica y la gestión institucional estatal y de la sociedad civil. No busca ser meta, punto de llegada, al contrario, es una propuesta que invita permanentemente al debate, la discusión teórica, la reflexión, al examen de las formas de hacer y al pensamiento crítico. Dr. Yuri Pérez Martíne