Creating an HLA-homozygous iPS cell bank for the Brazilian population: Challenges and opportunities.

Identifying human leukocyte antigen (HLA) haplotype-homozygous donors for the generation of induced pluripotent stem (iPS) cell lines permits the construction of biobanks immunologically compatible with significant numbers of individuals for use in therapy. However, two questions must be addressed to create such a bank: how many cell lines are necessary to match most of the recipient population and how many people should be tested to find these donors? In Japan and the UK, 50 and 100 distinct HLA-A, -B, and -DRB1 triple-homozygous haplotypes would cover 90% of those populations, respectively. Using data from the Brazilian National Registry of Bone Marrow Donors (REDOME), encompassing 4,017,239 individuals, we identified 1,906 distinct triple-homozygous HLA haplotypes. In Brazil, 559 triple-homozygous cell lines cover 95% of the population, and 3.8 million people would have to be screened. Finally, we show the contribution of the 30 most frequent triple-homozygous HLA haplotypes in Brazil to populations of different countries.

Zika Virus Strains and Dengue Virus Induce Distinct Proteomic Changes in Neural Stem Cells and Neurospheres.

Brain abnormalities and congenital malformations have been linked to the circulating strain of Zika virus (ZIKV) in Brazil since 2016 during the microcephaly outbreak; however, the molecular mechanisms behind several of these alterations and differential viral molecular targets have not been fully elucidated. Here we explore the proteomic alterations induced by ZIKV by comparing the Brazilian (Br ZIKV) and the African (MR766) viral strains, in addition to comparing them to the molecular responses to the Dengue virus type 2 (DENV). Neural stem cells (NSCs) derived from induced pluripotent stem (iPSCs) were cultured both as monolayers and in suspension (resulting in neurospheres), which were then infected with ZIKV (Br ZIKV or ZIKV MR766) or DENV to assess alterations within neural cells. Large-scale proteomic analyses allowed the comparison not only between viral strains but also regarding the two- and three-dimensional cellular models of neural cells derived from iPSCs, and the effects on their interaction. Altered pathways and biological processes were observed related to cell death, cell cycle dysregulation, and neurogenesis. These results reinforce already published data and provide further information regarding the biological alterations induced by ZIKV and DENV in neural cells.

Influence of diode laser and warm air drying on the shear bond strength of Lithium di-silicate to Dentin. An in-vitro study / Influência do laser de diodo e da secagem com ar quente na resistência ao cisalhamento do Di-silicato de Lítio a dentina. Um estudo in-vitro

Objective: The current study aimed to evaluate the effect of diode laser irradiation (980 nm) and warm air drying (50°C) on shear bond strength between Lithium di-silicate (IPS e.max; Ivoclar) and human dentin using both (Etch & Rinse) adhesive, Adper™ Single Bond 2 (3M ESPE) and (Self-etch) adhesive, Single Bond UniversalTM(3M ESPE) before adhesive polymerization. Material and Methods: 54 sound lower molars were sectioned to obtain flat dentinal surfaces. Specimens were divided into 2 equal groups (n=27): Group 1 (ER) and Group 2 (SE) according to bonding approach. Each subgroup was subdivided according to dentin surface treatment into 3 equal subgroups (n=9): Control (Co), Diode laser irradiation (L) and Warm air drying (W). All specimens were adhesively cemented to IPS e.max® CAD discs using RelyX™ Ultimate Clicker™(3M ESPE) resin cement. Samples were then subjected to pre-loading in a thermodynamic manner. All samples were tested for shear bond strength using computer-controlled material testing machine. Data analysis was performed using two-way (ANOVA) (p< 0.05) followed by pair-wise Tukey's post-hoc tests. Results: In (SE) group, the subgroup (W) had the highest shear bond strength values followed by (Co) subgroup and the least was (L) subgroup with statistically significant difference. As for (ER) group, the subgroup (W) had the highest shear bond strength values followed by (Co) subgroup and the least was (L) subgroup with no statistically significant difference. Conclusion: Warm air drying for (SE) bonding approach increased shear bond strength of Lithium di-silicate to human dentin and can be introduced as a new effective protocol.(AU)

Objetivo: o objetivo do estudo atual é avaliar o efeito da radiação do laser de diodo (980nm) e secagem de ar quente (50°C) na resistência ao cisalhamento entre dissilicati de Lítio (IPS e.max; Ivoclar) e a dentina humana usando ambos modelos de adesivos (condicionamento total) Adper™ Single Bond e (auto-condicionante) Single Bond Universal™ (3M ESPE), Single Bond UniversalTM (3M ESPE) antes da fotopolimerização. Material e Métodos: 54 segundos molares inferiores foram selecionados para obter superfícies dentinárias planas. Os espécimes foram divididos em 2 grupos iguais (n=27): grupo 1 (ER) e grupo 2 (SE) de acordo com protocolo de adesividade. Cada grupo foi subdividido de acordo com o tratamento de superfície dentro de 3 subgrupos iguais (n=9): Controle (co), irradiação com laser de diodo (L) e secagem com ar quente (W). Todos os espécimes foram adesivamente cimentados a discos de IPS emax CAD usando RelyX Ultimate Clicker (3M ESPE) cimento resinoso. As amostras foram então submetidas a pré-carregamento de forma termodinâmica. Todas as amostras foram testadas para resistência a cisalhamento usando máquina de teste de materiais controlados por computador. A análise de dados foi realizada usando ANOVA dois fatores (p<0.05) seguindo por testes de Tukey pareado como test post-hoc. Resultados: No grupo (SE), o subgrupo (W) apresentou maiores valores de resistência ao cisalhamento seguindo o subgrupo (Co) é o menor foi o subgrupo (L) com diferença estatisticamente significativa. Já para o grupo (Er), o subgrupo (W) apresentou os maiores valores de resistência ao cisalhamento seguido do grupo (Co) e o menor foi o subgrupo (L) sem diferença estatisticamente significante. Conclusão: a secagem com ar quente para a abordagem de adesividade (Se) aumentou a resistência ao cisalhamento do dissilicato de lítio à dentina humana e poderia ser introduzido como um novo e eficaz protocolo(AU)

Non-permissive SARS-CoV-2 infection in human neurospheres.

Coronavirus disease 2019 (COVID-19) was initially described as a viral infection of the respiratory tract. It is now known, however, that several other organs are affected, including the brain. Neurological manifestations such as stroke, encephalitis, and psychiatric conditions have been reported in COVID-19 patients, but the neurotropic potential of the virus is still debated. Herein, we sought to investigate SARS-CoV-2 infection in human neural cells. We demonstrated that SARS-CoV-2 infection of neural tissue is non-permissive, however, it can elicit inflammatory response and cell damage. These findings add to the hypothesis that most of the neural damage caused by SARS-CoV-2 infection is due to a systemic inflammation leading to indirect harmful effects on the central nervous system despite the absence of local viral replication.

An evaluation of the effect of different veneer materials on fracture resistance of zirconia core / Avaliação dos diferentes efeitos de materiais de revestimento na resistência à fratura do núcleo de zircônia

Objectives: this in vitro study was done to evaluate the effect of packable P60 composite and Tetric N-Ceram composite veneer material on fracture strength of zirconia cores. Material and Methods:Twenty four zirconium cores (Vita, Germany) with 0.7 mm thickness were fabricated by CAD/CAM technology and then subjected to air abrasion with 50 µm of Al2O3. Cores were randomly divided into three groups according to veneering material (group A: control group sandblasted with 50 µm Al2O3 veneered by IPS E-max Ceram porcelain, group C: sandblasted with 50 µm Al2O3, etching with hydrofluoric acid and veneered with P60 composite, group E: sandblasted with 50 µm Al2O3, etching with hydrofluoric acid and veneered with Tetric N-Ceram composite). All crowns were subjected to fracture strength test in the testing machine, with load application by steel ball indenter and 0.5 mm/min. cross head speed. Results: statistical analysis was carried out utilizing one-way ANOVA, LSD. The results of fracture strength value test showed the highest mean value was registered for group (A), and the lowest mean for group (E). One-way ANOVA test represented that, there was a statistically high significant different among all groups. LSD results showed a high significant difference increase in fracture resistance for Group A at p value (*p < 0.001 High significant). Conclusions: Within the limitation of this study, sandblasting zirconia core with 50 µm Al2O3 and veneering with conventional ceramic produced restoration with acceptable fracture resistance value (AU)

Objetivo: o objetivo desse estudo in vitro foi avaliar o efeito dos compósitos P60 compactáveis e material laminado de compósitos Tetric-N Ceram na resistência à fratura de núcleos de zircônia. Material e métodos:Vinte e quatro núcleos de zircônia (Vita, Germany) com 0.7 mm de espessura fabricados por tecnologia de CAD/CAM e sinterizados em alta temperatura (1450ºC por 60 min) de acordo com as instruções do fabricante. Núcleos totalmente de zircônias foram submetidos a abração com 50 µm de Al2O3. Os núcleos de zircônia foram divididos de forma randomiza em três grupos de acordo com o material de revestimento (grupo A: grupo controle jateado com 50 µm Al2O3 folheado com porcelana IPS E-max Ceram, grupo C: jateado com 50 µm Al2O3 , coberto com concentração 9,5% de ácido fluorídrico e folheado com resina composta compatível, grupo E: jateado com 50 µm Al2O3 coberto com concentração 9,5% de ácido fluorídrico e folheado com porcelana IPS E-max Ceram. Todos os espécimes (coroas) foram submetidos a teste de resistência a fratura com máquina de teste universal, essa máquina utilizou para aplicação de carga bola de aço com 6mm de diâmetro 0,5 mm/min de velocidade da cruzeta. Resultados: análise estatística foi realizada utilizando One-way ANOVA, LSD. O resultado da resistência a fratura mostra que o maior valor médio foi registrado no grupo A e a menor média para o grupo E. O resultado do teste One-Way ANOVA representaram que houve uma diferença estatisticamente alta e significante entre todos os grupos. LSD foi executada para mostrar a fonte de significância. Os resultados do LSD mostraram um aumento da diferença significativa alta na resistência à fratura para o grupo A no valor de p (p < 0.001 signicancia alta). Conclusão: Dentro das limitações desse estudo o uso convencional de jateamento com 50 µm Al2O3 e estratificação com cerâmica convencional produziu restauração com valor aceitável de resistência a fratura (AU)

Mechanical and thermal stress analysis of hybrid ceramic and lithium disilicate based ceramic CAD-CAM inlays using 3-D finite element analysis / Análise do estresse mecânico e térmico de inlays de cerâmica hibrida e de cerâmica a base de dissilicato de lítio através da análise em 3D de elementos finitos

Objective: The aim of this study was to analyze mechanical and thermal stresses of hybrid ceramic and lithium disilicate based ceramic of CAD/CAM inlays using 3D Finite element analysis. Material and Methods:A three dimensions finite element model of permanent maxillary premolar designed according to standard anatomy with class II cavity preparation for inlay restored with two different ceramic materials: 1- Hybrid ceramic (Vita Enamic), 2- Lithium disilicate based ceramic (IPS e.max CAD). Totally six runs were performed on the model as: One loading case for each restorative material was tested in stress analysis; seven points of loading with 140N vertically applied at palatal cusp tip and cusp slop, marginal ridges and central fossa while the models base was fixed as a boundary condition in the two cases. Two thermal analysis cases were performed for each restoration material by applying 5ºC and 55ºC on the crown surface including the restoration surface. Results:The results of all structures were separated from the rest of the model to analyze the magnitude of stress in each component. For each group, maximum stresses on restorative materials, cement, enamel, and dentin were evaluated separately. Both ceramic materials generated similar stress distribution patterns for all groups when a total occlusal load of 140 N was applied. Conclusion: Thermal fluctuations of temperature have a great influence on the stresses induced on both restoration and tooth structure. IPS e.max CAD produced more favorable stresses on the tooth structure than Vita Enamic. (AU)

Objetivo: O objetivo deste estudo foi analisar o estresse mecânico e térmico de inlays de cerâmica hibrida e a base de dissilicato de lítio através da análise em 3D de elementos finitos. Material e Métodos: Foi realizado o design de um modelo de três dimensões em elementos finitos de um pré-molar superior de acordo com os padrões anatômicos de um preparo de cavidade classe II para restauração de inlay em dois tipos de cerâmicas diferentes: 1 Cerâmica hibrida (Vita Enamic), 2 ­ Cerâmica a base de dissilicato de lítio (IPS e.max CAD). No total, seis execuções foram realizadas no modelo como: Uma carga para cada material restaurador foi testado para análise de tensão; sete pontos de carga com 140N foi aplicado verticalmente na ponta da cúspide palatina, saliência da cúspide, cristas marginais e fossa central enquanto a base do modelo foi fixada como uma condição de limite nos dois casos. Duas análises térmicas foram realizadas para cada material de restauração, aplicando 5ºC e 55ºC na superfície da coroa, incluindo a superfície da restauração. Resultados: Os resultados de todas as estruturas foram separados do resto do modelo para analisar a magnitude do estresse de cada componente. Para cada grupo, o máximo de estresse nos materiais restauradores, cemento, esmalte e dentina foram avaliados separadamente. Padrões similares de distribuição de estresse foram gerados em todos os grupos para ambos os materiais cerâmicos, quando a carga oclusal de 140 N foi aplicada. Conclusão: As variações térmicas de temperatura têm uma grande influência nas tensões induzidas na restauração e na estrutura dentária. IPS e.max CAD produziu tensões mais favoráveis na estrutura dentária do que o Vita Enamic (AU)

One-year clinical evaluation of IPS Empress CAD versus polished Celtra Duo ceramic Laminate veneers (randomized controlled clinical trial) / Avaliação clínica de um ano de IPS Empress CAD versus laminados de cerâmica Celtra Duo polidos (ensaio clínico randomizado controlado)

Objective: To evaluate fracture resistance and survival rate of IPS Empress CAD versus Polished Celtra Duo ceramic laminate veneers. Material and Methods:Thirty-six ceramic laminate veneers were fabricated for maxillary anterior teeth. The patients were divided into two groups according to the material Group 1(control group) fabricated from IPS Empress CAD laminate veneers and group 2 (intervention group) fabricated from Polished Celtra Duo laminate veneers. Standardized the same preparation with butt joint design and chamfer finish line located supra gingival were performed for all the teeth. The fabrication of the veneers was performed using Cad\Cam (Ceramill motion) machine, with software (Exocad). The veneers surfaces were treated and silanated according to the manufacture instruction of each ceramic and enamel surfaces were etched where total etch adhesive protocol was obeyed using BISCO. Follow up sessions were done every two months up to one year for each patient using dental probe and operator vision to evaluate the fracture, survival rate, marginal adaptation, sensitivity and caries. according to USPHS criteria (United States Public Health Service). This was performed by an experienced, blinded investigator. Results: Fracture resistance, marginal adaptation, retention, caries and sensitivity were evaluated according to the criteria of USPHS and we found there is no significant difference as both groups scaled zero score. Conclusion: Both IPS Empress Cad and Polished Celtra Duo laminate veneers revealed successful clinical performance in terms of fracture resistance, marginal adaptation, retention, and sensitivity after one year follow up period (AU)

Objetivo: Avaliar a resistência à fratura e a taxa de sobrevivência de laminados de cerâmica IPS Empress CAD versus Celtra Duo polidos. Material e Métodos: Trinta e seis facetas cerâmicas laminadas foram confeccionadas para dentes anteriores superiores. Os pacientes foram divididos em dois grupos de acordo com o material Grupo 1 (grupo controle) confeccionado com laminados IPS Empress CAD e grupo 2 (grupo intervenção) confeccionado com laminados Celtra Duo polidos. A mesma forma de preparo e acabamento de chanfro localizado supra-gengival padronizados foram realizados para todos os dentes. A confecção das facetas foi realizada em máquina Cad \ Cam (Ceramill motion), com software (Exocad). As superfícies laminadas foram tratadas e silanizadas de acordo com as instruções do fabricante de cada cerâmica e as superfícies de esmalte foram condicionadas seguindo o protocolo adesivo de condicionamento ácido total com BISCO. Sessões de acompanhamento foram realizadas a cada dois meses durante um ano para cada paciente usando sonda exploradora e visão do operador para avaliar a fratura, taxa de sobrevivência, adaptação marginal, sensibilidade e cárie; de acordo com os critérios USPHS (Serviço de Saúde Pública dos Estados Unidos). Isso foi realizado por um investigador experiente e cego. Resultados: a resistência à fratura, adaptação marginal, retenção, cárie e sensibilidade foram avaliadas de acordo com os critérios da USPHS e não encontramos diferença significativa, pois ambos os grupos escalonaram pontuação zero. Conclusão: As facetas laminadas IPS Empress Cad e Celtra Duo polido revelaram desempenho clínico bem-sucedido em termos de resistência à fratura, adaptação marginal, retenção e sensibilidade após um período de acompanhamento de um ano. (AU)

Probing structural properties and antioxidant activity mechanisms for eleocarpanthraquinone.

In this work, we present a computational investigation on the structure and energetics of eleocarpanthraquinone, a newly isolated polyphenolic anthrone-antraquinone. Properties such as bond lengths, angles, atomic charges, bond dissociation enthalpies (BDEs), and ionization potential (IP) were determined through the use of density functional theory (DFT). The B3LYP and M06-2X exchange-correlation functionals were employed along with the 6-31+G(d,p), 6-31+ +G(d,p), and 6-311+G(d,p) basis sets for performing computations in the gas-phase, water, methanol, and ethanol. The conformation presenting all the hydroxyl groups undergoing hydrogen-bond interactions with neighboring oxygen atoms (conformation 5) was assigned as the most stable structure while its counterpart presenting no hydrogen-bond interaction was found to be 36.45 kcal/mol less stable than conformation 5 in the potential energy surface probed at the B3LYP/6-311+G(d,p) level of theory in the gas-phase, for instance. More importantly, the lowest O-H bond dissociation enthalpy was determined to be 93.80 kcal/mol at the B3LYP/6-311+G(d,p) level of theory in water against the 146.58 kcal/mol regarding the IP computed at the same approach, suggesting the hydrogen atom transfer mechanism as being preferred over the single electron transfer mechanism in regards to the antioxidant potential for the case of eleocarpanthraquinone; the same conclusion was drawn from the outcomes of all the other approaches used.

Interaction of fibroblasts and induced pluripotent stem cells with poly(vinyl alcohol)-based hydrogel substrates.

Induced pluripotent stem cells (iPSCs) provide a promising means of creating custom-tailored cell lines for cellular therapies. Their application in regenerative medicine, however, depends on the possibility that the maintenance and differentiation of cells and organs occur under defined conditions. One major component of stem cell culture systems is the substrate, where the cells must attach and proliferate. The present study aimed to investigate the putative cytotoxic effects of poly(vinyl alcohol) (PVA)-based matrices on the in vitro culture of mouse fetal fibroblasts. In addition, the PVA-based hydrogels were used to determine the capacity of bovine induced pluripotent stem cells (biPSCs) to adhere and proliferate on synthetic substrates. Our results show that both cell types interacted with the substrate and presented proliferation during culture. The biPSCs formed new colonies when cell suspensions were placed onto the hydrogel surface for culture. These results may represent a new characterized xeno-free clinical grade culture system to be widely applied in cell-based therapies.

A platform to understand amyotrophic lateral sclerosis (ALS) and extend human motor neurons longevity / Plataforma para empezar a comprender la esclerosis lateral amiotrófica (ELA) y extender la longevidad de neuronas motoras humanas

In vitro modeling of neurodegenerative diseases is now possible by using patient-derived induced pluripotent stem cells (iPS). Through them, it is nowadays conceivable to obtain human neurons and glia, and study diseases cellular and molecular mechanisms, an attribute that was previously unavailable to any human condition. Amyotrophic lateral sclerosis (ALS) is one of the diseases that has gained a rapid advance with iPS technology. By differentiating motor neurons from iPS cells of ALS- patients, we are studying the mechanisms underlying ALS- disease onset and progression. Here, we introduce a cellular platform to help maintain longevity of ALS iPS-motor neurons, a cellular feature relevant for most late-onset human diseases. Long term cultures of patient-derived iPS cells might prove to be critical for the development of personalized-drugs.

Actualmente es posible modelar in vitro enfermedades neurodegenerativas humanas mediante el uso de células madre pluripotentes inducidas (iPS) derivadas del paciente. A través de ellas, es hoy concebible obtener neuronas y glía humanas, y estudiar mecanismos celulares y moleculares de enfermedades, un atributo que anteriormente no era posible para ninguna condición humana. La esclerosis lateral amiotrófica (ELA) es una de las enfermedades que se ha beneficiado con la tecnología de iPS. Al diferenciar neuronas motoras de células iPS obtenidas de pacientes con ELA, hemos iniciado estudios sobre los mecanismos que subyacen a la aparición y progresión de la enfermedad. Aquí, presentamos el desarrollo de una plataforma celular que permite extender la longevidad de las neuronas motoras derivadas de iPS, una característica relevante para la mayoría de las enfermedades humanas de inicio tardío. Los cultivos a largo plazo de células iPS provenientes de pacientes pueden ser determinantes en el desarrollo de terapias asociadas a la medicina de precisión.

Adult and iPS-derived non-parenchymal cells regulate liver organoid development through differential modulation of Wnt and TGF-ß.

BACKGROUND: Liver organoid technology holds great promises to be used in large-scale population-based drug screening and in future regenerative medicine strategies. Recently, some studies reported robust protocols for generating isogenic liver organoids using liver parenchymal and non-parenchymal cells derived from induced pluripotent stem cells (iPS) or using isogenic adult primary non-parenchymal cells. However, the use of whole iPS-derived cells could represent great challenges for a translational perspective. METHODS: Here, we evaluated the influence of isogenic versus heterogenic non-parenchymal cells, using iPS-derived or adult primary cell lines, in the liver organoid development. We tested four groups comprised of all different combinations of non-parenchymal cells for the liver functionality in vitro. Gene expression and protein secretion of important hepatic function markers were evaluated. Additionally, liver development-associated signaling pathways were tested. Finally, organoid label-free proteomic analysis and non-parenchymal cell secretome were performed in all groups at day 12. RESULTS: We show that liver organoids generated using primary mesenchymal stromal cells and iPS-derived endothelial cells expressed and produced significantly more albumin and showed increased expression of CYP1A1, CYP1A2, and TDO2 while presented reduced TGF-ß and Wnt signaling activity. Proteomics analysis revealed that major shifts in protein expression induced by this specific combination of non-parenchymal cells are related to integrin profile and TGF-ß/Wnt signaling activity. CONCLUSION: Aiming the translation of this technology bench-to-bedside, this work highlights the role of important developmental pathways that are modulated by non-parenchymal cells enhancing the liver organoid maturation.

Transplantation studies reveal internuclear transfer of toxic RNA in engrafted muscles of myotonic dystrophy 1 mice.

BACKGROUND: Stem cell transplantation represents a potential therapeutic option for muscular dystrophies (MD). However, to date, most reports have utilized mouse models for recessive types of MD. Here we performed studies to determine whether myotonic dystrophy 1 (DM1), an autosomal dominant type of MD, could benefit from cell transplantation. METHODS: We injected human pluripotent stem (PS) cell-derived myogenic progenitors into the muscles of a novel mouse model combining immunodeficiency and skeletal muscle pathology of DM1 and investigated transplanted mice for engraftment as well as for the presence of RNA foci and alternative splicing pattern. FINDINGS: Engraftment was clearly observed in recipient mice, but unexpectedly, we detected RNA foci in donor-derived engrafted myonuclei. These foci proved to be pathogenic as we observed MBNL1 sequestration and abnormal alternative splicing in donor-derived transcripts. INTERPRETATION: It has been assumed that toxic CUG repeat-containing RNA forms foci in situ in the nucleus in which it is expressed, but these data suggest that CUG repeat-containing RNA may also exit the nucleus and traffic to other nuclei in the syncytial myofiber, where it can exert pathological effects. FUND: This project was supported by funds from the LaBonte/Shawn family and NIH grants R01 AR055299 and AR071439 (R.C.R.P.). R.M-G. was funded by CONACyT-Mexico (#394378).

Evaluation of the antioxidant potential of myricetin 3-O-α-L-rhamnopyranoside and myricetin 4'-O-α-L-rhamnopyranoside through a computational study.

In this work, we present a computational study on the antioxidant potential of myricetin 3-O-α-L-rhamnopyranoside (Compound M3) and myricetin 4'-O-α-L-rhamnopyranoside (Compound M4'). Structural parameters, bond dissociation enthalpies (BDEs), ionization potentials (IPs), proton dissociation enthalpies (PDEs), proton affinities (PAs), and electron transfer enthalpies (ETEs), which are properties connected with different mechanisms related to antioxidant activity, were determined using density functional theory (DFT) with B3LYP, LC-ωPBE, M06-2X, and BMK functionals along with the 6-311G(d,p) and 6-311+G(d,p) basis sets in the gas phase, water, and pentylethanoate. The values obtained were compared with results previously available in the literature for myricetin (the parent molecule and a well-known antioxidant) and myricetin 3,4'-di-O-α-L-rhamnopyranoside (Compound M3,4'). As the BDEs are considerably lower than the IPs, the HAT mechanism is preferred over SET for the compounds M3 and M4'. The present study indicates Compound M3 as having its lowest bond dissociation enthalpy from the several different OH groups with similar value to the lowest for myricetin (74.72 kcal/mol versus 74.8 kcal/mol, respectively, at the B3LYP/6-311G(d,p) level of theory in the gas phase) and, thus, presenting antioxidant potential as good as its parent molecule. On the other hand, Compound M4' presented 78.97 kcal/mol as the lowest BDE at the B3LYP/6-311G(d,p) level of theory in the gas phase, that is very close to the 78.34 kcal/mol computed using the same approach for Compound M3,4'. Therefore, the present investigation indicated Compound M4' as being a slightly inferior antioxidant (with antioxidant potential comparable to Compound M3,4') than Compound M3. In addition, the inclusion of the sugar moiety studied here in the position 4'-ArOH of myricetin seems to have a more marked impact (downward) on the antioxidant activity than the glycosylation in the position 3-ArOH.

Caracterización de la atención psicoterapéutica en la IPS CES Sabaneta, Colombia, 2014-2015 / Characterization of psychotherapeutic assistance in the IPS CES Sabaneta, Colombia, 2014-2015

Resumen Los cambios en la cosmovisión imperante en los últimos decenios han traído consigo sujetos con nuevas características, intereses y exigencias, que han impactado a la psicología clínica en tanto le exigen nuevas formas de comprender e intervenir las problemáticas demandadas por ellos en contextos institucionales, entre otros. De cara a estas nuevas realidades y con el fin de hacer más pertinentes las intervenciones psicoterapéuticas, se realizó una caracterización de las mismas en la Institución Prestadora de Servicios de Salud -IPS- CES Sabaneta (Colombia) durante los años 2014-201 5, a través de un análisis descriptivo de 9.140 registros de atención y 532 historias clínicas elegidas aleatoriamente. Se estudiaron aspectos sociodemográficos, administrativos y teórico-técnicos de la atención a partir de análisis de frecuencia, medidas de tendencia central, de dispersión y de forma. Se encontró que consultan tres hombres por cada mujer, siendo la población entre los 6 y 18 años la que tiene el mayor peso (59,77%). Los trastornos emocionales y del comportamiento fueron el principal diagnóstico (44,92%), seguido por los trastornos neuróticos secundarios a situaciones estresantes y somatomorfos (18,6%). Llama la atención la baja prevalencia de los trastornos de personalidad (0,56%) y que el 48,9% de los pacientes asistió a un máximo de seis sesiones. Los resultados permiten ajustar los perfiles y estrategias de atención de acuerdo con las problemáticas más prevalentes, así como los procesos administrativos y formativos relacionados con ellas.

Abstract The changes happened in the prevailing worldview in the last decades have brought subjects with new characteristics, interests and requirements, which have had an impact on the clinical psychology, as a consequence, they require from clinical psychology new ways of understanding and controlling the arisen issues that concern the institutional contexts. In order to face these realities, relevant psychotherapeutic interventions were implemented through a characterization of health attendance carried out in IPS CES Saba-neta during 2014-201 5, conducted by a descriptive analysis of 9140 records of assistance and an analysis of 532 clinical records, chosen randomly. Sociodemographic, administrative and theoretical-technical aspects of the care service were studied based on frequency analysis, measures of central tendency and dispersion using the SPSS software. It was found that 3 men in 1 woman attend to medical service, being the population between ages of 6 and 18 the one that present the highest weight (59, 77 %). Emotional and behavioral disorders were the main diagnosis (44.92%), followed by neurotic disorders secondary to stressful and somatomorphic situations (18.6%). It is noticed the low prevalence of personality disorders (0.56%) even though 48.9% of patients attended a maximum of six sessions. The results allow adjusting the profiles and care service strategies according to the most prevalent problems, as well as the administrative and formative processes that involve these issues.

Distinct features of rabbit and human adipose-derived mesenchymal stem cells: implications for biotechnology and translational research.

INTRODUCTION: Owing to their similarity with humans, rabbits are useful for multiple applications in biotechnology and translational research from basic to preclinical studies. In this sense, mesenchymal stem cells (MSCs) are known for their therapeutic potential and promising future in regenerative medicine. As many studies have been using rabbit adipose-derived MSCs (ASCs) as a model of human ASCs (hASCs), it is fundamental to compare their characteristics and understand how distinct features could affect the translation to human medicine. OBJECTIVE: The aim of this study was to comparatively characterize rabbit ASCs (rASCs) and hASCs to further uses in biotechnology and translational studies. MATERIALS AND METHODS: rASCs and hASCs were isolated and characterized by their immunophenotype, differentiation potential, proliferative profile, and nuclear stability in vitro. RESULTS AND DISCUSSION: Both ASCs presented differentiation potential to osteocytes, chondrocytes, and adipocytes and shared similar immunophenotype expression to CD105+, CD34-, and CD45-, but rabbit cells expressed significantly lower CD73 and CD90 than human cells. In addition, rASCs presented greater clonogenic potential and proliferation rate than hASCs but no difference in nuclear alterations. CONCLUSION: The distinct features of rASCs and hASCs can positively or negatively affect their use for different applications in biotechnology (such as cell reprogramming) and translational studies (such as cell transplantation, tissue engineering, and pharmacokinetics). Nevertheless, the particularities between rabbit and human MSCs should not prevent rabbit use in preclinical models, but care should be taken to interpret results and properly translate animal findings to medicine.

Exogenous human OKSM factors maintain pluripotency gene expression of bovine and porcine iPS-like cells obtained with STEMCCA delivery system.

OBJECTIVES: The use of induced pluripotent stem (iPS) cells as an alternative to embryonic stem cells to produce transgenic animals requires the development of a biotechnological platform for their generation. In this study, different strategies for the generation of bovine and porcine iPS cells were evaluated. Lentiviral vectors were used to deliver human factors OCT4, SOX2, KLF4 and c-MYC (OKSM) into bovine and porcine embryonic fibroblasts and different culture conditions were evaluated. RESULTS: Protocols based on the integrative lentiviral vector STEMCCA produced porcine iPS-like cells more efficiently than in bovine cells. The iPS-like cells generated displayed stem cell features; however, expression of exogenous factors was maintained along at least 12 passages. Since inactivation of the exogenous factors is still a major bottleneck for establishing fully reprogrammed iPS cells, defining culture conditions that support endogenous OKSM expression is critical for the efficient generation of farm animals' iPS cells.

Alternativas de tratamientos de superficie para adhesión de cerámica de disilicado de litio / Alternatives of surface treatments for adhesion of lithium disilicate ceramics

Introduction: the clinical success of a restoration is strongly associated with the quality and durability of the ceramic-cement resin interface. In order to obtain an adequate union between these materials of different nature surface treatments are used and achieve mechanical retention or chemical interaction. Objectives: to check if any method promotes a true chemical bond between lithium disilicate ceramics and resin cement. As well as determineif there is any treatment that reports bonding values comparable to hydrofluoric acid and silane (gold standard). Methods: a systematic literature review was developed based on the PRISMA strategy, where the databases were searched: Science Direct, Pubmed (MEDLINE), EMBASE, Springer Journal, SciELO with MeSH and free terms from 2005 to November 2016 for articles in English and Spanish on surface treatments for lithium disilicate. Results: from 58 publications selected a sample of 21 articles. Two articles reported high risk of bias. Conclusions: hydrofluoric acid and silane continue to be the method with the highest and most reliable adhesion values in the literature. Universal adhesives are an alternative to promote chemical adhesion additional to the silane. Diamond burs, Nd: YAG and Er: YAG laser are not recommended as surface treatments(AU)

Introducción: el éxito clínico de una restauración se asocia fuertemente a la calidad y duración de la interface cerámica-cemento resinoso. Para que exista una adecuada unión entre estos materiales de distinta naturaleza se emplean tratamientos de superficie para lograr una buena retención mecánica o interacción química. Objetivos: revisar si algún método promueve una verdadera adhesión química entre la cerámica de disilicato de litio y el cemento resinoso, así como determinar si existe algún tratamiento que reporte valores de unión comparables al ácido fluorhídrico y silano (patrón de oro). Métodos: se desarrolló una revisión sistemática de literatura basada en la estrategia PRISMA, donde se buscó en las bases de datos: Science Direct, Pubmed (MEDLINE), EMBASE, Springer Journal, SciELO con términos MeSH y libres desde el 2005 a noviembre de 2016 para artículos en inglés y español sobre tratamientos de superficie para disilicato de litio. Resultados: de 58 publicaciones, se seleccionó una muestra de 21 artículos. Dos artículos reportaron riesgo de sesgo alto. Conclusiones: el ácido fluorhídrico y silano continúan siendo el método con los valores de adhesión más altos y confiables de la literatura. Los adhesivos universales son una alternativa para promover adhesión química adicional al silano. Fresas diamantadas, laser Nd: YAG y Er:YAG no se recomienda como tratamientos de superficie(AU)

Alternativas de tratamientos de superficie para adhesión de cerámica de disilicado de litio / Alternatives of surface treatments for adhesion of lithium disilicate ceramics

Introduction: the clinical success of a restoration is strongly associated with the quality and durability of the ceramic-cement resin interface. In order to obtain an adequate union between these materials of different nature surface treatments are used and achieve mechanical retention or chemical interaction. Objectives: to check if any method promotes a true chemical bond between lithium disilicate ceramics and resin cement. As well as determineif there is any treatment that reports bonding values comparable to hydrofluoric acid and silane (gold standard). Methods: a systematic literature review was developed based on the PRISMA strategy, where the databases were searched: Science Direct, Pubmed (MEDLINE), EMBASE, Springer Journal, SciELO with MeSH and free terms from 2005 to November 2016 for articles in English and Spanish on surface treatments for lithium disilicate. Results: from 58 publications selected a sample of 21 articles. Two articles reported high risk of bias. Conclusions: hydrofluoric acid and silane continue to be the method with the highest and most reliable adhesion values in the literature. Universal adhesives are an alternative to promote chemical adhesion additional to the silane. Diamond burs, Nd: YAG and Er: YAG laser are not recommended as surface treatments(AU)

Introducción: el éxito clínico de una restauración se asocia fuertemente a la calidad y duración de la interface cerámica-cemento resinoso. Para que exista una adecuada unión entre estos materiales de distinta naturaleza se emplean tratamientos de superficie para lograr una buena retención mecánica o interacción química. Objetivos: revisar si algún método promueve una verdadera adhesión química entre la cerámica de disilicato de litio y el cemento resinoso, así como determinar si existe algún tratamiento que reporte valores de unión comparables al ácido fluorhídrico y silano (patrón de oro). Métodos: se desarrolló una revisión sistemática de literatura basada en la estrategia PRISMA, donde se buscó en las bases de datos: Science Direct, Pubmed (MEDLINE), EMBASE, Springer Journal, SciELO con términos MeSH y libres desde el 2005 a noviembre de 2016 para artículos en inglés y español sobre tratamientos de superficie para disilicato de litio. Resultados: de 58 publicaciones, se seleccionó una muestra de 21 artículos. Dos artículos reportaron riesgo de sesgo alto. Conclusiones: el ácido fluorhídrico y silano continúan siendo el método con los valores de adhesión más altos y confiables de la literatura. Los adhesivos universales son una alternativa para promover adhesión química adicional al silano. Fresas diamantadas, laser Nd: YAG y Er:YAG no se recomienda como tratamientos de superficie(AU)

Modeling Inflammation in Autism Spectrum Disorders Using Stem Cells.

Recent reports show an increase in the incidence of Autism Spectrum Disorders (ASD) to 1 in every 59 children up to 8 years old in 11 states in North America. Induced pluripotent stem cell (iPSC) technology offers a groundbreaking platform for the study of polygenic neurodevelopmental disorders in live cells. Robust inflammation states and immune system dysfunctions are associated with ASD and several cell types participate on triggering and sustaining these processes. In this review, we will examine the contribution of neuroinflammation to the development of autistic features and discuss potential therapeutic approaches. We will review the available tools, emphasizing stem cell modeling as a technology to investigate the various molecular pathways and different cell types involved in the process of neuroinflammation in ASD.

Células-tronco pluripotentes induzidas (células iPS) em animais domésticos e a possibilidade de geração in vitro de gametas / Induced pluripotent stem cells (iPS cells) in domestic animals and the possibility to generate gametes in vitro

Células-tronco são conhecidas pela característica de alto potencial de auto-renovação e podem ser classificadas segundo seu estágio de indiferenciação e perfil epigenético. Células-tronco embrionárias (CTEs) e células-tronco adultas são bastante estudadas e caracterizadas, principalmente nos modelos humano e em camundongos por apresentarem novas possibilidades tanto para a medicina regenerativa quanto para o entendimento do desenvolvimento inicial dos mamíferos. Contudo, a derivação e a manutenção de CTEs em espécies domésticas é desafiadora e apresenta resultados inconsistentes em relação à manutenção da pluripotência in vitro. Nesse contexto, a geração das células pluripotentes induzidas in vitro (células iPS ou iPSCs) é imprescindível para a geração de novas possibilidades na medicina veterinária regenerativa e reprodutiva devido à capacidade de diferenciação destas em uma variedade de outros tipos celulares, inclusive em animais. Assim, esta revisão apresenta a geração das células iPS em animais domésticos, e em especial, recentes estudos sobre as etapas necessárias para a possibilidade de geração de gametas funcionais in vitro, uma importante contribuição na correção de infertilidades, conservação de espécies ameaçadas de extinção e geração de indivíduos geneticamente superiores ou modificados para aplicações agropecuárias ou biomédicas.(AU)

Stem cells are widely known for their high potential for self-renewal, being classified according to their stage of undifferentiation and epigenetic profile. Embryonic stem cells (ES) and adult stem cells are already well studied and characterized, mainly in human and mouse models, once they present new possibilities both for regenerative medicine and for understanding the initial development of mammals. However, the derivation and maintenance of ES in domestic species is challenging and presents inconsistent results regarding maintenance of in vitro pluripotency. In this context, the derivation of in vitro induced pluripotent cells (iPS cells or iPSCs) enables new possibilities in regenerative and reproductive veterinary medicine due to their ability to differentiate into a variety of other cell types. Therefore, this review presents the generation of iPS cells in domestic animals, and focuses on recent studies on the steps necessary for the generation of functional gametes in vitro, an important contribution of stem cells aiming the correction of infertility, the conservation of species in risk of extinction and for the generation of genetically superior or modified organisms for agricultural and biomedical applications.(AU)