3D culture of mesenchymal stem cells from the yolk sac to generate intestinal organoid.

Organoids are in vitro models that originated from the three-dimensional culture of stem cells with the ability to reproduce part of the in vivo structural and functional specificities of body organs. Intestinal organoids have great relevance in cell therapy, as they provide more accurate information about tissue composition and architecture in relation to two-dimensional culture, in addition to serving as a study model for host interaction and drug testing. The yolk sac (YS) is a promising source of mesenchymal stem cells (MSCs), which are multipotent stem cells with self-renewal ability and potential to differentiated into mesenchymal lineages. Besides this, the YS is responsible for the formation of intestinal epithelium during embryonic development. Thus, the aim of this study was to verify if the three-dimensional in vitro culture of stem cells derived from the canine YS is capable of developing intestinal organoids. MSCs from the canine YS and gut cells were isolated and characterized, then three-dimensionally cultured in Matrigel. In both cells lineages, spherical organoids were observed and after 10 days the gut cells formed crypt-like buds and villus-like structures. Despite having the same induction of differentiation process and having the expression of intestinal markers, the MSC from the YS morphology was not in the form of crypt budding. The hypothesis is that these cells could generate structures equivalent to the intestinal organoids of the colon that other studies showed formed only spherical structures. The culture of MSC from the YS, as well as the establishment of protocols for 3D cultivation of this tissue, is relevant, as it will serve as a tool in various applications in basic and scientific biology.

Ozone therapy: protocol for treating canine parvovirus infection.

Canine Parvovirus infection is a disease caused by Canine Parvovirus (CPV) that results in hemorrhagic gastroenteritis and secondary infections, mainly in puppies between six weeks and six months old that are not immunized. Since there is no specific treatment for the condition, supportive therapy based on antibiotics, antiemetics, and non-steroidal anti-inflammatory drugs is traditionally used. Ozone therapy is an economical treatment that has bactericidal, fungicidal, and antiviral properties, besides promoting oxygenation and tissue regeneration, as well as anti-inflammatory and analgesic effects, and was used as a complementary therapy in this study. Therefore, four mixed-breed dogs, aged between 2 and 3 months, with no previous immunization against CPV and testing positive for the virus in a rapid test were selected. The animals were randomly distributed into two groups, being 1: the control group (n=2) that received only supportive treatment; and 2: the experimental group (n=2), that in addition to conventional therapy received intravenously 500 mL of ozonized Ringer's Lactate solution. Before treatment and after 24 and 48 hours, the following clinical signs were evaluated: episodes of emesis and diarrhea, weight, hydration, blood glucose level, abdominal pain, and blood count. One control group animal died within the first hours of hospitalization. Both animals in the experimental group presented faster resolution of diarrheal episodes and shorter hospitalization time when compared to the surviving animal that received only supportive treatment. Although further studies are needed, ozone therapy showed promising results for the treatment of canine parvovirus.

A Parvovirose canina é uma doença infecciosa causada pelo Parvovírus Canino (CPV) que resulta em gastroenterite hemorrágica e infecções secundárias, principalmente em cachorros entre seis semanas e seis meses de idade não imunizados. Como não existe tratamento específico para a doença, utiliza-se tradicionalmente terapia de suporte baseada em antibióticos, antieméticos, e anti-inflamatórios não esteroides. A Ozonioterapia é um tratamento econômico com propriedades bactericidas, fungicidas e antivirais, além de promover a oxigenação e a regeneração dos tecidos, efeitos anti-inflamatórios e analgésicos, e foi utilizada como terapia complementar neste estudo. Neste estudo, foram selecionados quatro cães sem raça definida, com idades entre 2 e 3 meses, sem imunização prévia contra CPV, que testaram positivo para o vírus em um teste rápido. Os animais foram distribuídos aleatoriamente em dois grupos, sendo 1: o grupo controle (n=2) que recebeu apenas tratamento de suporte; e 2: o grupo experimental (n=2), que além da terapia convencional recebeu por via intravenosa 500 mL de Lactato de Ringer ozonizado. Antes do tratamento, após 24 e 48 horas, foram avaliados os seguintes sinais clínicos: episódios de êmese e diarreia, peso, hidratação, glicemia, dores abdominais, e hemograma. Um animal do grupo controle foi a óbito nas primeiras horas de internação. Ambos os animais do grupo experimental apresentaram uma resolução mais rápida dos episódios de diarreia e um tempo de internação mais curto quando comparado com o animal sobrevivente que recebeu apenas tratamento de suporte. Embora sejam necessários mais estudos, a ozonoterapia demonstrou resultados promissores para o tratamento do parvovírus canino.

Potential application of aminiotic stem cells in veterinary medicine.

In regenerative medicine stem cell biology has become one of the most interesting and more often studied subject. The amniotic membrane is the innermost layer of the fetal membranes and is considered a potential tool to treat many pathologies. It is used because it can be collected from discarded fetal material and is a rich source of stem cells with high proliferation and plasticity ratio capable of proliferating and differentiate in vitro. We propose to elucidate the characteristics and potencial clinical application of cells derived of amniotic membrane in veterinary medicine.

Low intensity training provokes adaptations on muscle fibrosis of a muscular dystrophy model / El entrenamiento de baja intensidad provoca adaptaciones en la fibrosis muscular de un modelo de distrofia muscular

Duchenne muscular dystrophy (DMD) is a genetic neuromuscular disorder with progressive clinical signs until death, around the second decade of life. Mdx is the most used animal model to pre-clinical studies of DMD. Parameters of exercise on this muscular disease are still unknown. This research aimed to investigate if the low intensity treadmill training would exacerbate the markers of muscle injury, fibrosis, and the composition of the extracellular matrix by type I and III collagens of the mdx model. Dystrophic 11-week-old male mice were separated in exercised (mdxE, n=8) and sedentary (mdxC, n=8) groups. Wild-type mice were used as control (WT, n=8). Exercised group underwent a LIT protocol (9 m/min, 30min, 3days/week, 60 days) on a horizontal treadmill. Gastrocnemius muscle was collected at day 60 and processed to morphological and morphometric analyzes. Sedentary mdx animals presented inflammatory infiltrate and necrotic fibers. Histochemical analysis revealed that the perimysium of the mdxC group is organized into thick and clustered collagen fibers, which generates a larger area of intramuscular collagen fibers for these animals. Histomorphometry attested that fraction area of collagen fibers of mdxC group was higher than mdxE group (p=0.04) and mdxE group values similar to WT group (p=1.00). Centrally located nuclei fibers and the variance coefficient (VC) of minimal Feret's diameter was similar in mdxE and mdxC groups (p=1.00) and both groups presented higher mean values than WT group (p<0.00). Immunohistochemistry revealed the presence of type I collagen mainly in the mdxC group. LIT protocol had not exacerbated muscle injuries resulting from the dystrophindeficiency membrane fragility at the same time that had reduced the intramuscular collagen deposition. LIT had positively influenced these markers of dystrophic muscle injury on gastrocnemius muscle of mdx model.

La distrofia muscular de Duchenne (DMD) es un trastorno neuromuscular genético con signos clínicos progresivos hasta la muerte, alrededor de la segunda década de la vida. Mdx es el modelo animal más utilizado para estudios preclínicos de DMD. Los parámetros del ejercicio en esta enfermedad muscular aún se desconocen. Esta investigación tuvo como objetivo investigar si el entrenamiento de cinta de baja intensidad exacerbaría los marcadores de lesión muscular, fibrosis y la composición de la matriz extracelular por colágenos tipo I y III del modelo mdx. Ratones machos de 11 semanas de edad con distrofia se separaron en grupos ejercitados (mdxE, n = 8) y sedentarios (mdxC, n = 8). Se usaron ratones de tipo salvaje como control (WT, n = 8). El grupo sometido a ejercicio se sometió a un protocolo LIT (9 m / min, 30 min, 3 días / semana, 60 días) en una cinta de trotar horizontal. El músculo gastrocnemio se retiró el día 60 y se procesó para realizar análisis morfológicos y morfométricos. Los animales sedentarios mdx presentaron infiltrado inflamatorio y fibras necróticas. El análisis histoquímico reveló que el perimisio del grupo mdxC está organizado en fibras de colágeno gruesas y agrupadas, lo que genera una mayor área de fibras de colágeno intramusculares para estos animales. La histomorfometría indicó que el área de fracción de las fibras de colágeno del grupo mdxC era más alta que el grupo mdxE (p = 0,04) y los valores del grupo mdxE eran similares al grupo WT (p = 1,00). Las fibras de los núcleos ubicados centralmente y el coeficiente de varianza (VC) del diámetro mínimo de Feret fueron similares en los grupos mdxE y mdxC (p = 1,00) y ambos grupos presentaron valores medios más altos que el grupo WT (p <0,00). La inmunohistoquímica reveló la presencia de colágeno tipo I principalmente en el grupo mdxC. El protocolo LIT no había agravado las lesiones musculares resultantes de la fragilidad de la membrana con deficiencia de distrofina al mismo tiempo que había reducido la deposición intramuscular de colágeno. LIT ha influido positivamente en estos marcadores de lesión muscular distrófica en el músculo gastrocnemio del modelo mdx.

Associação entre o desenvolvimento neuropsicomotor e fatores de risco biológico e ambientais em crianças na primeira infância / Association between neuropsychomotor development and biological and environmental risk factors in early childhood children

RESUMO Objetivo: avaliar a associação entre o desenvolvimento infantil e riscos biológicos e ambientais. Métodos: foram selecionadas 30 crianças (0 a 30 meses), residentes em uma cidade no Vale do Jequitinhonha, Minas Gerais, Brasil, atendidas pelo Centro Viva Vida de Referência Secundária. Os critérios de inclusão foram: crianças que tinham história clínica de prematuridade e/ou desnutrição moderada à grave e outros fatores neonatais de risco. As crianças foram avaliadas quanto ao desenvolvimento por meio do teste Denver II e a qualidade de estímulo no ambiente domiciliar foi avaliada pelo Home Observation for Measurement of the Environment (HOME). Resultados: das 30 crianças avaliadas, 60% dos ambientes foram considerados de risco para o desenvolvimento infantil e 43,3% apresentou desenvolvimento inadequado. O principal domínio afetado foi o da linguagem. Maior escolaridade materna, constituição familiar biparental apresentaram associação com o adequado desenvolvimento infantil. A presença de intercorrências neonatais e necessidade de internação no centro de terapia intensiva foram mais presentes nas crianças que falharam no teste. A menor receptividade dos pais e disponibilidade de materiais para aprendizagem no domicílio foram fatores associados ao pior desempenho das crianças no Denver II. Conclusão: as crianças de alto risco do presente estudo apresentaram atraso no desenvolvimento, especialmente no domínio linguagem. Esses atrasos estão associados à baixa escolaridade materna, à relação monoparental, receptividade dos pais e intercorrências neonatais.

ABSTRACT Objective: to verify the association between infant development and biological and environmental risks. Methods: 30 children between 0-30 months, living in a town in Minas Gerais, Brazil, attending a Health Center, were selected. The inclusion criteria were children who had a history of prematurity and/or, moderate to severe malnutrition and other risk factors. Their development was assessed through the Denver II test and the quality of stimulation in the home environment assessed by the Home Observation for Measurement of the Environment (HOME). Results: 60% of the environments were considered to be at risk for child development and 43.3% presented inadequate development. The main domain affected was the language. Higher maternal education and bi-parental families showed a relationship with proper child development. Neonatal complications and hospitalization in intensive care units were more common in children who failed the test. The parents' low receptivity and the availability of materials at home were factors associated with the children's worst development performance. Conclusion: the results show that the high-risk children in this study had a developmental delay, especially in the language area. These delays are associated with low maternal education, single-parent home, parents' responsiveness and neonatal complications.

Growth and development and their environmental and biological determinants / Crescimento e desenvolvimento e seus determinantes ambientais e biológicos

Abstract Objective To investigate child growth, cognitive/language development, and their environmental and biological determinants. Methods This was a cross-sectional, predictive correlation study with all 92 children aged 24-36 months who attended the municipal early childhood education network in a town in the Vale do Jequitinhonha region, in 2011. The socioeconomic profile was determined using the questionnaire of the Associação Brasileira de Empresas de Pesquisa. The socio-demographicand maternal and child health profiles were created through a self-prepared questionnaire. The height-for-age indicator was selected to represent growth. Cognitive/language development was assessed through the Bayley Scale of Infant and Toddler Development. The quality of educational environments was assessed by Infant/Toddler Environment Scale; the home environment was assessed by the Home Observation for Measurement of the Environment. The neighborhood quality was determined by a self-prepared questionnaire. A multivariate linear regression analysis was performed. Results Families were predominantly from socioeconomic class D, with low parental education. The prevalence of stunted growth was 14.1%; cognitive and language development were below average at 28.6% and 28.3%, respectively. Educational institutions were classified as inadequate, and 69.6% of homes were classified as presenting a risk for development. Factors such as access to parks and pharmacies and perceived security received the worst score regarding neighborhood environment. Biological variables showed a greater association with growth and environmental variables with development. Conclusion The results showed a high prevalence of stunting and below-average results for cognitive/language development among the participating children. Both environmental and biological factors were related to growth and development. However, biological variables showed a greater association with growth, whereas environmental variables were associated with development.

Resumo Objetivo Investigar o crescimento e desenvolvimento cognitivo/linguagem de crianças e seus determinantes ambientais e biológicos. Método Estudo transversal, correlacional preditivo, com todas as 92 crianças entre 24-36 meses, frequentadoras da rede municipal de educação infantil de uma cidade no Vale do Jequitinhonha, ano 2011. Traçou-se o perfil econômico utilizando-se o questionário da Associação Brasileira de Empresas de Pesquisa. O perfil sociodemográfico e saúde materno-infantil por questionário próprio. Elegeu-se o indicador estatura/idade para representar o crescimento. O desenvolvimento cognitivo/linguagem foi avaliado por meio do Bayley Scale of Infant and Toddler Development. Avaliou-se os ambientes educacionais pelo Infant/Toddler Environment Scale, e o ambiente domiciliar pelo Home Observation for Measurement of the Environment. Aferiu-se a qualidade da vizinhança através de questionário próprio. Foram realizadas análises de regressão linear multivariada. Resultados As famílias eram predominantemente da classe D com baixa escolaridade dos pais. A prevalência de déficit de estatura foi 14,1%; desenvolvimento abaixo da média na linguagem 28,6% e cognitivo 28,3%. As instituições educacionais classificaram-se como inadequadas e 69,6% dos domicílios como de risco para o desenvolvimento. Aspectos como, disponibilidade de praças e farmácias e segurança foram aspectos de pior pontuação no ambiente vizinhança. Variáveis biológicas demonstraram maior associação com o crescimento e variáveis ambientais ao desenvolvimento. Conclusão Observou-se elevado déficit de estatura e de resultados abaixo da média para desenvolvimento cognitivo/linguagem entre as crianças participantes. Fatores ambientais e biológicos relacionaram-se tanto ao crescimento quanto ao desenvolvimento. Entretanto, variáveis biológicas demonstraram maior associação com o crescimento e variáveis ambientais com o desenvolvimento.

Growth and development and their environmental and biological determinants.

OBJECTIVE: To investigate child growth, cognitive/language development, and their environmental and biological determinants. METHODS: This was a cross-sectional, predictive correlation study with all 92 children aged 24-36 months who attended the municipal early childhood education network in a town in the Vale do Jequitinhonha region, in 2011. The socioeconomic profile was determined using the questionnaire of the Associação Brasileira de Empresas de Pesquisa. The socio-demographicand maternal and child health profiles were created through a self-prepared questionnaire. The height-for-age indicator was selected to represent growth. Cognitive/language development was assessed through the Bayley Scale of Infant and Toddler Development. The quality of educational environments was assessed by Infant/Toddler Environment Scale; the home environment was assessed by the Home Observation for Measurement of the Environment. The neighborhood quality was determined by a self-prepared questionnaire. A multivariate linear regression analysis was performed. RESULTS: Families were predominantly from socioeconomic class D, with low parental education. The prevalence of stunted growth was 14.1%; cognitive and language development were below average at 28.6% and 28.3%, respectively. Educational institutions were classified as inadequate, and 69.6% of homes were classified as presenting a risk for development. Factors such as access to parks and pharmacies and perceived security received the worst score regarding neighborhood environment. Biological variables showed a greater association with growth and environmental variables with development. CONCLUSION: The results showed a high prevalence of stunting and below-average results for cognitive/language development among the participating children. Both environmental and biological factors were related to growth and development. However, biological variables showed a greater association with growth, whereas environmental variables were associated with development.