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1.
Hepatol Commun ; 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-34558820

RESUMO

The only definitive therapy for end-stage liver disease is whole-organ transplantation. The success of this intervention is severely limited by the complexity of the surgery, the cost of patient care, the need for long-term immunosuppression, and the shortage of donor organs. In rodents and humans, end-stage degeneration of hepatocyte function is associated with disruption of the liver-specific transcriptional network and a nearly complete loss of promoter P1-driven hepatocyte nuclear factor 4-alpha (P1-HNF4α) activity. Re-expression of HNF4α2, the predominant P1-HNF4α, reinstates the transcriptional network, normalizes the genes important for hepatocyte function, and reverses liver failure in rodents. In this study, we tested the effectiveness of supplementary expression of human HNF4α2 messenger RNA (mRNA) in primary human hepatocytes isolated from explanted livers of patients who underwent transplant for end-stage irreversibly decompensated liver failure (Child-Pugh B, C) resulting from alcohol-mediated cirrhosis and nonalcoholic steatohepatitis. Re-expression of HNF4α2 in decompensated cirrhotic human hepatocytes corrects the disrupted transcriptional network and normalizes the expression of genes important for hepatocyte function, improving liver-specific protein expression. End-stage liver disease in humans is associated with both loss of P1-HNF4α expression and failure of its localization to the nucleus. We found that while HNF4α2 re-expression increased the amount of P1-HNF4α protein in hepatocytes, it did not alter the ability of hepatocytes to localize P1-HNF4α to their nuclei. Conclusion: Re-expression of HNF4α2 mRNA in livers of patients with end-stage disease may be an effective therapy for terminal liver failure that would circumvent the need for organ transplantation. The efficacy of this strategy may be enhanced by discovering the cause for loss of nuclear P1-HNF4α localization in end-stage cirrhosis, a process not found in rodent studies.

2.
Edumecentro ; 13(3): 64-80, jul.-sept. 2021. tab
Artigo em Espanhol | LILACS | ID: biblio-1278989

RESUMO

RESUMEN Fundamento: los programas de preparación para asumir una jubilación saludable son elementos clave en la protección de la calidad de vida de las personas prejubilables. Objetivo: mostrar la eficacia de la aplicación de un programa educativo dirigido a personas en etapa de prejubilación para el afrontamiento de una vida futura con calidad. Métodos: se realizó un estudio polietápico con diseño cuasiexperimental, en el Policlínico Universitario "Marta Abreu", en Santa Clara, Villa Clara, entre enero 2017-febrero 2019. Se utilizaron métodos teóricos: histórico-lógico, análisis-síntesis, inductivo-deductivo y sistémico-estructural; empíricos: análisis documental, cuestionario antes y después de aplicado el programa; y para valorar la propuesta se utilizaron los grupos focal y nominal, y los criterios de especialistas, además se emplearon métodos matemático-estadísticos. Resultados: el diagnóstico realizado demostró la necesidad de la preparación de las personas prejubilables para afrontar con calidad una nueva etapa de sus vidas, por lo que se diseñó un programa educativo el cual fue estructurado en cinco módulos: la jubilación como una etapa trascendental en la vida del adulto, salud física y salud psicológica, bienestar psicosocial e inteligencia emocional, planificación y emprendimiento y la adaptación integral a una nueva fase. Fue valorado por especialistas, quienes lo consideraron muy pertinente y viable. Conclusiones: el programa educativo resultó efectivo para el mejoramiento de la calidad de vida en personas prejubilables del grupo estudio, pues se constataron cambios significativos en sentido de avance al comparar sus conocimientos antes y después de aplicado y en relación con el grupo de control.


ABSTRACT Background: the preparation programs for assuming a healthy retirement are key elements in protecting life quality in early retirement people. Objective: to show the effectiveness of the application of an educational program aimed at people in pre-retirement stage to face a future life with quality. Methods: a multi-stage study with a quasi-experimental design was carried out at the "Marta Abreu" University Polyclinic, in Santa Clara, Villa Clara, from January 2017 to February 2019. Theoretical methods were used: historical-logical, analysis-synthesis, inductive-deductive and systemic-structural; empirical ones: documentary analysis, questionnaire before and after applying the program; and to assess the proposal, the focal and nominal groups were used, as well as the criteria of specialists, in addition, mathematical-statistical methods were used. Results: the diagnosis showed the need to prepare early retirees to face a new stage of their lives with quality, so an educational program was designed which was structured in five modules: retirement as a transcendental stage in the adult life, physical and psychological health, psychosocial well-being and emotional intelligence, planning and entrepreneurship, and comprehensive adaptation to a new phase. It was valued by specialists, who considered it very relevant and feasible. Conclusions: the educational program was effective for improving the quality of life in early retirement in the study group, since meaningful changes in the direction of progress were found when comparing their knowledge before and after it was applied and in relation to the control group.


Assuntos
Qualidade de Vida , Estratégias de Saúde , Educação Médica , Cursos de Capacitação , Aprendizagem
3.
Polymers (Basel) ; 13(16)2021 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-34451306

RESUMO

A green, effective methodology for the preparation of water-based dispersions of poly(lactic acid) (PLA) for coating purposes is herein presented. The procedure consists of two steps: in the first one, an oil-in-water emulsion is obtained by mixing a solution of PLA in ethyl acetate with a water phase containing surfactant and stabilizer. Different homogenization methods as well as oil/water phase ratio, surfactant and stabilizer combinations were screened. In the second step, the quantitative evaporation of the organic provides water dispersions of PLA that are stable, at least, over several weeks at room temperature or at 4 °C. Particle size was in the 200-500 nm range, depending on the preparation conditions, as confirmed by scanning electron microscope (SEM) analysis. PLA was found not to suffer significant molecular weight degradation by gel permeation chromatography (GPC) analysis. Furthermore, two selected formulations with glass transition temperature (Tg) of 51 °C and 34 °C were tested for the preparation of PLA films by drying in PTFE capsules. In both cases, continuous films that are homogeneous by Fourier-transform infrared spectroscopy (FT-IR) and SEM observation were obtained only when drying was performed above 60 °C. The formulation with lower Tg results in films which are more flexible and transparent.

4.
Foods ; 10(7)2021 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-34359460

RESUMO

In this work, novel active films based on ethylene vinyl alcohol copolymer (EVOH) and cinnamaldehyde (CIN) were successfully obtained employing a hybrid technique consisting of a two-step protocol involving the preparation of a polymeric EVOH-CIN masterbatch by solvent-casting for its further utilization in the preparation of bioactive EVOH-based films by melt extrusion processing. The influence of CIN over the EVOH matrix was studied in terms of optical, morphological, thermal, and mechanical properties. Optically transparent films were obtained and the incorporation of cinnamaldehyde resulted in yellow-colored films, producing a blocking effect in the UV region. A decrease in the glass transition temperature was observed in the formulations containing cinnamaldehyde, indicating a plasticizing effect. This phenomenon was confirmed by an increase in the elongation at break values of the extruded films. Results from thermogravimetric analysis determined a slight decrease in the thermal stability of EVOH provoked by the vaporization of the bioactive compound. Bioactive properties of the films were also studied; the presence of residual cinnamaldehyde in EVOH after being subjected to an extrusion process conferred some radical scavenging activity determined by the DPPH assay whereas films were able to exert antifungal activity in vapor phase against Penicillium expansum. Therefore, the present work shows the potential of the hybrid technique employed in this study for the preparation of bioactive films by a ready industrial process technology for food packaging applications.

5.
Mol Cell ; 81(16): 3323-3338.e14, 2021 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-34352207

RESUMO

The emerging "epitranscriptomics" field is providing insights into the biological and pathological roles of different RNA modifications. The RNA methyltransferase METTL1 catalyzes N7-methylguanosine (m7G) modification of tRNAs. Here we find METTL1 is frequently amplified and overexpressed in cancers and is associated with poor patient survival. METTL1 depletion causes decreased abundance of m7G-modified tRNAs and altered cell cycle and inhibits oncogenicity. Conversely, METTL1 overexpression induces oncogenic cell transformation and cancer. Mechanistically, we find increased abundance of m7G-modified tRNAs, in particular Arg-TCT-4-1, and increased translation of mRNAs, including cell cycle regulators that are enriched in the corresponding AGA codon. Accordingly, Arg-TCT expression is elevated in many tumor types and is associated with patient survival, and strikingly, overexpression of this individual tRNA induces oncogenic transformation. Thus, METTL1-mediated tRNA modification drives oncogenic transformation through a remodeling of the mRNA "translatome" to increase expression of growth-promoting proteins and represents a promising anti-cancer target.


Assuntos
Carcinogênese/genética , Metiltransferases/genética , Neoplasias/genética , tRNA Metiltransferases/genética , Guanosina/análogos & derivados , Guanosina/genética , Humanos , Metilação , Neoplasias/patologia , Oncogenes/genética , Processamento Pós-Transcricional do RNA/genética , RNA Mensageiro/genética , RNA de Transferência/genética
6.
Artigo em Inglês | MEDLINE | ID: mdl-34323642

RESUMO

RNA aptamers are single-stranded nucleic acids of 20-100 nucleotides, with high sensitivity and specificity against particular molecular targets. In vitro production and selection of aptamers can be performed using the SELEX method. However, this procedure requires considerable time and cost. In this sense, bioinformatics tools play an important role in reducing the time and cost associated with development and production of aptamers. In this article, we propose bioinformatics strategies for modeling and analysis of the interaction with molecular targets for two RNA aptamers: ATP binding RNA aptamer and iSpinach aptamer. For this purpose, molecular modeling of the tertiary structure of the aptamers was performed with two servers (SimRNA and RNAComposer); and AutoDock Vina and rDock programs were used to dock their respective ligands. The predictions developed with these methods could be used for in silico design of RNA aptamers, through a simple and accessible methodology.Supplemental data for this article is available online at https://doi.org/10.1080/15257770.2021.1951754 .

7.
Biotechnol Prog ; : e3187, 2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34164947

RESUMO

Protein concentration determination is a necessary in-process control for the downstream operations within biomanufacturing. As production transitions from batch mode to an integrated continuous bioprocess paradigm, there is a growing need to move protein concentration quantitation from off-line to in-line analysis. One solution to fulfill this process analytical technology need is an in-line index of refraction (IoR) sensor to measure protein concentration in real time. Here the performance of an IoR sensor is evaluated through a series of experiments to assess linear response, buffer matrix effects, dynamic range, sensor-to-sensor variability, and the limits of detection and quantitation. The performance of the sensor was also tested in two bioprocessing scenarios, ultrafiltration and capture chromatography. The implementation of this in-line IoR sensor for real-time protein concentration analysis and monitoring has the potential to improve continuous bioprocess manufacturing.

8.
Exp Biol Med (Maywood) ; : 15353702211009228, 2021 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-33957803

RESUMO

Metabolic syndrome is a complex disease that involves multiple organ systems including a critical role for the liver. Non-alcoholic fatty liver disease (NAFLD) is a key component of the metabolic syndrome and fatty liver is linked to a range of metabolic dysfunctions that occur in approximately 25% of the population. A panel of experts recently agreed that the acronym, NAFLD, did not properly characterize this heterogeneous disease given the associated metabolic abnormalities such as type 2 diabetes mellitus (T2D), obesity, and hypertension. Therefore, metabolic dysfunction-associated fatty liver disease (MAFLD) has been proposed as the new term to cover the heterogeneity identified in the NAFLD patient population. Although many rodent models of NAFLD/NASH have been developed, they do not recapitulate the full disease spectrum in patients. Therefore, a platform has evolved initially focused on human biomimetic liver microphysiology systems that integrates fluorescent protein biosensors along with other key metrics, the microphysiology systems database, and quantitative systems pharmacology. Quantitative systems pharmacology is being applied to investigate the mechanisms of NAFLD/MAFLD progression to select molecular targets for fluorescent protein biosensors, to integrate computational and experimental methods to predict drugs for repurposing, and to facilitate novel drug development. Fluorescent protein biosensors are critical components of the platform since they enable monitoring of the pathophysiology of disease progression by defining and quantifying the temporal and spatial dynamics of protein functions in the biosensor cells, and serve as minimally invasive biomarkers of the physiological state of the microphysiology system experimental disease models. Here, we summarize the progress in developing human microphysiology system disease models of NAFLD/MAFLD from several laboratories, developing fluorescent protein biosensors to monitor and to measure NAFLD/MAFLD disease progression and implementation of quantitative systems pharmacology with the goal of repurposing drugs and guiding the creation of novel therapeutics.

9.
Semin Liver Dis ; 41(2): 213-224, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33992030

RESUMO

Chronic liver injury results in cirrhosis and end-stage liver disease (ESLD) which represents a leading cause of death worldwide, affecting people in their most productive years of life. Medical therapy can extend life, but the only definitive treatment is liver transplantation (LT). However, LT remains limited by access to quality donor organs and suboptimal long-term outcomes. The degeneration from healthy-functioning livers to cirrhosis and ESLD involves a dynamic process of hepatocyte damage, diminished hepatic function, and adaptation. However, the mechanisms responsible for deterioration of hepatocyte function and ultimately hepatic failure in man are poorly understood. We review the current understanding of cirrhosis and ESLD as a dynamic process and outline the current mechanisms associated with the development of hepatic failure from the clinical manifestations to energy adaptations, regeneration, and regulation of nuclear transcription factors. A new generation of therapeutics could target stabilization of hepatocyte differentiation and function to avoid the need for transplantation in patients with cirrhosis and ESLD.

10.
Philos Trans R Soc Lond B Biol Sci ; 376(1825): 20200168, 2021 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-33813884

RESUMO

Molluscan aquaculture is a major contributor to global seafood production, but is hampered by infectious disease outbreaks that can cause serious economic losses. Selective breeding has been widely used to improve disease resistance in major agricultural and aquaculture species, and has clear potential in molluscs, albeit its commercial application remains at a formative stage. Advances in genomic technologies, especially the development of cost-efficient genomic selection, have the potential to accelerate genetic improvement. However, tailored approaches are required owing to the distinctive reproductive and life cycle characteristics of molluscan species. Transgenesis and genome editing, in particular CRISPR/Cas systems, have been successfully trialled in molluscs and may further understanding and improvement of genetic resistance to disease through targeted changes to the host genome. Whole-organism genome editing is achievable on a much greater scale compared to other farmed species, making genome-wide CRISPR screening approaches plausible. This review discusses the current state and future potential of selective breeding, genomic tools and genome editing approaches to understand and improve host resistance to infectious disease in molluscs. This article is part of the Theo Murphy meeting issue 'Molluscan genomics: broad insights and future directions for a neglected phylum'.

11.
PLoS One ; 16(4): e0245898, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33798205

RESUMO

BACKGROUND: We aimed to compare the performance of the recent CASTLE score to J-CTO, CL and PROGRESS CTO scores in a comprehensive database of percutaneous coronary intervention of chronic total occlusion procedures. METHODS: Scores were calculated using raw data from 1,342 chronic total occlusion procedures included in REBECO Registry that includes learning and expert operators. Calibration, discrimination and reclassification were evaluated and compared. RESULTS: Mean score values were: CASTLE 1.60±1.10, J-CTO 2.15±1.24, PROGRESS 1.68±0.94 and CL 2.52±1.52 points. The overall percutaneous coronary intervention success rate was 77.8%. Calibration was good for CASTLE and CL, but not for J-CTO or PROGRESS scores. Discrimination: the area under the curve (AUC) of CASTLE (0.633) was significantly higher than PROGRESS (0.557) and similar to J-CTO (0.628) and CL (0.652). Reclassification: CASTLE, as assessed by integrated discrimination improvement, was superior to PROGRESS (integrated discrimination improvement +0.036, p<0.001), similar to J-CTO and slightly inferior to CL score (- 0.011, p = 0.004). Regarding net reclassification improvement, CASTLE reclassified better than PROGRESS (overall continuous net reclassification improvement 0.379, p<0.001) in roughly 20% of cases. CONCLUSION: Procedural percutaneous coronary intervention difficulty is not consistently depicted by available chronic total occlusion scores and is influenced by the characteristics of each chronic total occlusion cohort. In our study population, including expert and learning operators, the CASTLE score had slightly better overall performance along with CL score. However, we found only intermediate performance in the c-statistic predicting chronic total occlusion success among all scores.


Assuntos
Oclusão Coronária/cirurgia , Intervenção Coronária Percutânea , Idoso , Área Sob a Curva , Oclusão Coronária/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Medição de Risco , Índice de Gravidade de Doença , Resultado do Tratamento
12.
Curr Opin Gastroenterol ; 37(3): 224-230, 2021 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-33769378

RESUMO

PURPOSE OF REVIEW: In this review, we will explore recent advances in human induced pluripotent stem cell (iPSC)-based modeling of metabolic liver disease and biofabrication of synthetic human liver tissue while also discussing the emerging concept of synthetic biology to generate more physiologically relevant liver disease models. RECENT FINDING: iPSC-based platforms have facilitated the study of underlying cellular mechanisms and potential therapeutic strategies for a number of metabolic liver diseases. Concurrently, rapid progress in biofabrication and gene editing technologies have led to the generation of human hepatic tissue that more closely mimic the complexity of the liver. SUMMARY: iPSC-based liver tissue is rapidly becoming available for modeling liver physiology due to its ability to recapitulate the complex three-dimensional architecture of the liver and recapitulate interactions between the different cell types and their surroundings. These mini livers have also been used to recapitulate liver disease pathways using the tools of synthetic biology, such as gene editing, to control gene circuits. Further development in this field will undoubtedly bolster future investigations not only in disease modeling and basic research, but also in personalized medicine and autologous transplantation.

13.
Gigascience ; 10(3)2021 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-33764468

RESUMO

BACKGROUND: The Pacific oyster (Crassostrea gigas) is a bivalve mollusc with vital roles in coastal ecosystems and aquaculture globally. While extensive genomic tools are available for C. gigas, highly contiguous reference genomes are required to support both fundamental and applied research. Herein we report the creation and annotation of a chromosome-level assembly for C. gigas. FINDINGS: High-coverage long- and short-read sequence data generated on Pacific Biosciences and Illumina platforms were used to generate an initial assembly, which was then scaffolded into 10 pseudo-chromosomes using both Hi-C sequencing and a high-density linkage map. The assembly has a scaffold N50 of 58.4 Mb and a contig N50 of 1.8 Mb, representing a step advance on the previously published C. gigas assembly. Annotation based on Pacific Biosciences Iso-Seq and Illumina RNA-Seq resulted in identification of ∼30,000 putative protein-coding genes. Annotation of putative repeat elements highlighted an enrichment of Helitron rolling-circle transposable elements, suggesting their potential role in shaping the evolution of the C. gigas genome. CONCLUSIONS: This new chromosome-level assembly will be an enabling resource for genetics and genomics studies to support fundamental insight into bivalve biology, as well as for selective breeding of C. gigas in aquaculture.

14.
Medicentro (Villa Clara) ; 25(1): 92-106, ene.-mar. 2021. tab
Artigo em Espanhol | LILACS-Express | LILACS | ID: biblio-1287184

RESUMO

RESUMEN Introducción: la prejubilación es un proceso de relevancia social por la alta tasa de crecimiento de la población activa envejecida, cuya preocupación más relevante surge por la extensión, quizás sobredimensionada, de la vida laboral. Objetivo: determinar los resultados de la musicoterapia en personas prejubilables. Métodos: se realizó una intervención musicoterapéutica con el objetivo de regular las emociones en personas prejubilables, ante el dilema del cese del vínculo laboral o la continuidad de este; se realizó un estudio cuasi-experimental en el Policlínico Docente «Marta Abreu¼, Villa Clara, en el período de enero 2017 a febrero 2019. Se emplearon procedimientos, métodos y técnicas con aplicación de la musicoterapia en un grupo estudio de 200 personas prejubilables, de 55 a 65 años de edad. Se utilizaron métodos teóricos, empíricos y estadísticos-matemáticos. Se aplicaron técnicas psicológicas antes y después de la musicoterapia en el grupo estudio y en el grupo control. Resultados: se constataron resultados relevantes en el grupo estudio, y se registraron cambios muy significativos: más estados emocionales ansiosos bajo y depresivo leve, autoestima alta, y notable mejoría en la vulnerabilidad al estrés. En el grupo control no se reflejaron cambios favorables. Conclusiones: la musicoterapia resultó efectiva por el logro de cambios positivos en el estado emocional en personas prejubilables del grupo estudio. La identificación prometedora de potencialidades en el lenguaje musical, con la utilización mínima de la palabra en la atención integral a personas en etapa de prejubilación, constituyó una novedad.


ABSTRACT Introduction: early retirement is a process of social relevance due to the high growth rate of the aging workforce, whose most relevant concern arises from the extension, perhaps oversized, of working life. Objective: to determine the results of music therapy in early retired people. Methods: a music therapy intervention was carried out with the aim of regulating emotions in early retired people, faced with the dilemma of the termination of the employment relationship or its continuity; a quasi-experimental study was carried out at "Marta Abreu" Teaching Polyclinic, Villa Clara, from January 2017 to February 2019. Procedures, methods and techniques with application of music therapy were used in a study group of 200 early retired people aged 55 to 65 years. Theoretical, empirical and statistical-mathematical methods were used. Psychological techniques were applied in the study group before and after music therapy and in the control group without intervention. Results: relevant results were found in the study group, and very significant changes were recorded: more low anxiety and mild depressive emotional states, high self-esteem, and notable improvement in vulnerability to stress. No favorable changes were reflected in the control group. Conclusions: music therapy was effective for achieving positive changes in emotional state in early retired people from the study group. The promising identification of potentialities in musical language, with the minimum use of spoken language in comprehensive care for people in early retirement, was a novelty.

15.
BMC Genomics ; 22(1): 156, 2021 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-33676414

RESUMO

BACKGROUND: Salmon Rickettsial Syndrome (SRS), caused by Piscirickettsia salmonis, is one of the primary causes of morbidity and mortality in Atlantic salmon aquaculture, particularly in Chile. Host resistance is a heritable trait, and functional genomic studies have highlighted genes and pathways important in the response of salmon to the bacteria. However, the functional mechanisms underpinning genetic resistance are not yet well understood. In the current study, a large population of salmon pre-smolts were challenged with P. salmonis, with mortality levels recorded and samples taken for genotyping. In parallel, head kidney and liver samples were taken from animals of the same population with high and low genomic breeding values for resistance, and used for RNA-Sequencing to compare their transcriptome profile both pre and post infection. RESULTS: A significant and moderate heritability (h2 = 0.43) was shown for the trait of binary survival. Genome-wide association analyses using 38 K imputed SNP genotypes across 2265 animals highlighted that resistance is a polygenic trait. Several thousand genes were identified as differentially expressed between controls and infected samples, and enriched pathways related to the host immune response were highlighted. In addition, several networks with significant correlation with SRS resistance breeding values were identified, suggesting their involvement in mediating genetic resistance. These included apoptosis, cytoskeletal organisation, and the inflammasome. CONCLUSIONS: While resistance to SRS is a polygenic trait, this study has highlighted several relevant networks and genes that are likely to play a role in mediating genetic resistance. These genes may be future targets for functional studies, including genome editing, to further elucidate their role underpinning genetic variation in host resistance.


Assuntos
Doenças dos Peixes , Salmo salar , Animais , Doenças dos Peixes/genética , Estudo de Associação Genômica Ampla , Piscirickettsia , Salmo salar/genética , Análise de Sequência de RNA
17.
Cartilage ; 12(1): 102-111, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-30373376

RESUMO

OBJECTIVE: Human mesenchymal stem cells (hMSCs) are a promising source for regenerative medicine, especially mesodermal lineages. Clinical applications require an understanding of the mechanisms for transcriptional control to maintain the desired cell type. The aim of this study was to identify novel markers for differentiation of hMSCs into bone or cartilage with the use of Kartogenin, by RNA analysis using microarray technology, and explore the role of RhoA-Rho associated protein kinase (ROCK) inhibition in these. METHODS: Commercial human bone marrow derived primary mesenchymal stem cells were purchased from ATCC. Cells were differentiated in vitro in 2-dimensional cultures using Kartogenin as the main cartilage inducer and bone morphogenetic protein 2 for bone differentiation; cells were cultured with and without ROCK inhibitor Y-27632. After 21 days of culture, whole RNA was extracted and analyzed via Affimetrix microarrays. The most significant hits were validated by quantitative polymerase chain reaction. RESULTS: We found a total of 1,757 genes that were either up- or downregulated on differentiation, when compared to P1 hMSC (control) at day 0 of differentiation. Two members of the Serpin superfamily, SERPINA9 and SERPINB2, were significantly upregulated in the cartilage groups, whereas they were unchanged in the bone groups with and without ROCK inhibition. CONCLUSIONS: SERPINA9 and SERPINB2 are novel differentiation markers, and molecular regulator candidates for hMSC lineage commitment toward bone and cartilage, providing a new tool for regenerative medicine. Our study highlights the roles of these 2 genes, with significant upregulation of both in cell cultures stimulated with Kartogenin.

18.
Scand J Immunol ; 93(3): e13002, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33247472

RESUMO

Non-bilayer phospholipids arrangements (NPAs) are transient molecular associations different from lipid bilayers. When they become stable, they can trigger a disease in mice resembling human lupus, which is mainly characterized by the production of anti-NPA IgG antibodies. NPAs are stabilized on liposomes or cell bilayers by the drugs procainamide or chlorpromazine, which produce drug-induced lupus in humans. Here, we evaluated the participation of the TH 2 response, through its hallmark cytokine IL-4, on the development of the lupus-like disease in mice. Wild-type or IL-4 knockout BALB/c mice received liposomes bearing drug-induced NPAs, the drugs alone, or an anti-NPA monoclonal antibody (H308) to induce the lupus-like disease (the last two procedures stabilize NPAs on mice cells). IL-4 KO mice showed minor disease manifestations, compared to wild-type mice, with decreased production of anti-NPA IgG antibodies, no anti-cardiolipin, anti-histones and anticoagulant antibodies, and no kidney or skin lesions. In these mice, H308 was the only inducer of anti-NPA IgG antibodies. These findings indicate that IL-4 has a central role in the development of the murine lupus-like disease induced by NPA stabilization.


Assuntos
Interleucina-4/genética , Interleucina-4/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Fosfolipídeos/imunologia , Células Th2/imunologia , Animais , Anticorpos Monoclonais/imunologia , Autoanticorpos/imunologia , Modelos Animais de Doenças , Feminino , Imunoglobulina G/imunologia , Bicamadas Lipídicas/metabolismo , Lúpus Eritematoso Sistêmico/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout
19.
Pediatr Blood Cancer ; 68(1): e28719, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33026184

RESUMO

BACKGROUND/OBJECTIVES: While outcomes for pediatric T-cell acute lymphoblastic leukemia (T-ALL) are favorable, there are few widely accepted prognostic factors, limiting the ability to risk stratify therapy. DESIGN/METHODS: Dana-Farber Cancer Institute (DFCI) Protocols 05-001 and 11-001 enrolled pediatric patients with newly diagnosed B- or T-ALL from 2005 to 2011 and from 2012 to 2015, respectively. Protocol therapy was nearly identical for patients with T-ALL (N = 123), who were all initially assigned to the high-risk arm. End-induction minimal residual disease (MRD) was assessed by reverse transcription polymerase chain reaction (RT-PCR) or next-generation sequencing (NGS), but was not used to modify postinduction therapy. Early T-cell precursor (ETP) status was determined by flow cytometry. Cases with sufficient diagnostic DNA were retrospectively evaluated by targeted NGS of known genetic drivers of T-ALL, including Notch, PI3K, and Ras pathway genes. RESULTS: The 5-year event-free survival (EFS) and overall survival (OS) for patients with T-ALL was 81% (95% CI, 73-87%) and 90% (95% CI, 83-94%), respectively. ETP phenotype was associated with failure to achieve complete remission, but not with inferior OS. Low end-induction MRD (<10-4 ) was associated with superior disease-free survival (DFS). Pathogenic mutations of the PI3K pathway were mutually exclusive of ETP phenotype and were associated with inferior 5-year DFS and OS. CONCLUSIONS: Together, our findings demonstrate that ETP phenotype, end-induction MRD, and PI3K pathway mutation status are prognostically relevant in pediatric T-ALL and should be considered for risk classification in future trials. DFCI Protocols 05-001 and 11-001 are registered at www.clinicaltrials.gov as NCT00165087 and NCT01574274, respectively.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores Tumorais/genética , Neoplasia Residual/patologia , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patologia , Adolescente , Adulto , Criança , Pré-Escolar , Ensaios Clínicos Fase III como Assunto , Feminino , Seguimentos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Masculino , Neoplasia Residual/tratamento farmacológico , Neoplasia Residual/genética , Leucemia-Linfoma Linfoblástico de Células T Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células T Precursoras/genética , Prognóstico , Ensaios Clínicos Controlados Aleatórios como Assunto , Indução de Remissão , Estudos Retrospectivos , Taxa de Sobrevida , Adulto Jovem
20.
Nat Rev Gastroenterol Hepatol ; 18(4): 252-268, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33335282

RESUMO

Microphysiology systems (MPS), also called organs-on-chips and tissue chips, are miniaturized functional units of organs constructed with multiple cell types under a variety of physical and biochemical environmental cues that complement animal models as part of a new paradigm of drug discovery and development. Biomimetic human liver MPS have evolved from simpler 2D cell models, spheroids and organoids to address the increasing need to understand patient-specific mechanisms of complex and rare diseases, the response to therapeutic treatments, and the absorption, distribution, metabolism, excretion and toxicity of potential therapeutics. The parallel development and application of transdisciplinary technologies, including microfluidic devices, bioprinting, engineered matrix materials, defined physiological and pathophysiological media, patient-derived primary cells, and pluripotent stem cells as well as synthetic biology to engineer cell genes and functions, have created the potential to produce patient-specific, biomimetic MPS for detailed mechanistic studies. It is projected that success in the development and maturation of patient-derived MPS with known genotypes and fully matured adult phenotypes will lead to advanced applications in precision medicine. In this Review, we examine human biomimetic liver MPS that are designed to recapitulate the liver acinus structure and functions to enhance our knowledge of the mechanisms of disease progression and of the absorption, distribution, metabolism, excretion and toxicity of therapeutic candidates and drugs as well as to evaluate their mechanisms of action and their application in precision medicine and preclinical trials.


Assuntos
Biomimética , Desenvolvimento de Medicamentos , Fígado/metabolismo , Medicina de Precisão , Avaliação Pré-Clínica de Medicamentos/métodos , Humanos , Dispositivos Lab-On-A-Chip , Procedimentos Analíticos em Microchip , Microfluídica , Modelos Animais
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