16 research outputs found
Polimorfismos genéticos de enzimas del metabolismo de los folatos, drd4 y gstm1 y su relación con la disfunción temporomandibular
Temporomandibular disorder (TMD) is a multifactorial syndrome related to a critical period of human life. TMD has been associated with psychological dysfunctions, oxidative state and sexual dimorphism with coincidental occurrence along the pubertal development. In this work we study the association between TMD and genetic polymorphisms of folate metabolism, neurotransmission, oxidative and hormonal metabolism. Folate metabolism, which depends on genes variations and diet, is directly involved in genetic and epigenetic variations that can influence the changes of last growing period of development in human and the appearance of the TMD.
Methods
A case-control study was designed to evaluate the impact of genetic polymorphisms above described on TMD. A total of 229 individuals (69% women) were included at the study; 86 were patients with TMD and 143 were healthy control subjects. Subjects underwent to a clinical examination following the guidelines by the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Genotyping of 20 Single Nucleotide Polymorphisms (SNPs), divided in two groups, was performed by multiplex minisequencing preceded by multiplex PCR. Other seven genetic polymorphisms different from SNPs (deletions, insertions, tandem repeat, null genotype) were achieved by a multiplex-PCR. A chi-square test was performed to determine the differences in genotype and allelic frequencies between TMD patients and healthy subjects. To estimate TMD risk, in those polymorphisms that shown significant differences, odds ratio (OR) with a 95% of confidence interval were calculated.
Results
Six of the polymorphisms showed statistical associations with TMD. Four of them are related to enzymes of folates metabolism: Allele G of Serine Hydoxymethyltransferase 1 (SHMT1) rs1979277 (OR = 3.99; 95%CI 1.72, 9.25; p = 0.002), allele G of SHMT1 rs638416 (OR = 2.80; 95%CI 1.51, 5.21; p = 0.013), allele T of Methylentetrahydrofolate Dehydrogenase (MTHFD) rs2236225 (OR = 3.09; 95%CI 1.27, 7.50; p = 0.016) and allele A of Methionine Synthase Reductase (MTRR) rs1801394 (OR = 2.35; 95CI 1.10, 5.00; p = 0.037). An inflammatory oxidative stress enzyme, Gluthatione S-Tranferase Mu-1(GSTM1), null allele (OR = 2.21; 95%CI 1.24, 4.36; p = 0.030) and a neurotransmission receptor, Dopamine Receptor D4 (DRD4), long allele of 48 bp-repeat (OR = 3.62; 95%CI 0.76, 17.26; p = 0.161).
Conclusions
Some genetic polymorphisms related to folates metabolism, inflammatory oxidative stress, and neurotransmission responses to pain, has been significantly associated to TMD syndrom
Epithelial in vitro differentiation of human mesenchymal stem cells (hMSCs) from adipose tissue (AT) and bone marrow (BM): cellular characterization and study of HLA I and II expression
AGRADECIMIENTOS
Laboratorio de Citogenética del servicio de Análisis Clínicos
del Hospital Universitario Virgen de las Nieves. Servicio de Análisis Clínicos (Sección de Citometría/Biopatología tumoral) del Hos-
pital Universitario Virgen de las Nieves.Introducción: Las células troncales mesenquimales derivadas de tejido adiposo o médula ósea constituyen uno de
los tratamientos de terapia celular más utilizados en los ensayos clínicos actuales por su capacidad inmunomoduladora.
Además, por su potencial de diferenciación a células epiteliales pueden ser utilizadas en ingeniería tisular
incorporadas a tejidos artificiales como la piel o córnea, sustituyendo a las células epiteliales autólogas de estos
tejidos. Es necesario realizar una correcta caracterización de estas células diferenciadas y estudiar el efecto de la
diferenciación en la expresión del HLA de clase I y II.
Objetivos: Caracterizar y realizar los controles de calidad GMP en dos líneas de células mesenquimales troncales
humanas de distintos orígenes (tejido adiposo y médula ósea) tras diferenciarlas a células epiteliales in vitro, y
analizar si se modifica la expresión de los marcadores HLA I y II antes y después del proceso diferenciador.
Metodología: Se ha realizado el aislamiento y expansión de las dos líneas celulares de células mesenquimales
troncales a partir del tejido fuente y se ha procedido a su diferenciación in vitro a células epiteliales mediante
medios de cultivos suplementados con factores de crecimiento específico. Se han realizado controles de calidad
siguiendo los requerimientos de las normas de correcta fabricación y se ha estudiado por citometría de flujo la
expresión de HLA tipo I y II antes y después del proceso diferenciador. Finalmente se ha comprobado mediante
estudio histológico e inmunohistoquímico las características de las células diferenciadas.
Resultados: Se han aislado dos líneas de células mesenquimales troncales de tejido adiposo y médula ósea que
cumplen los controles de calidad propuestos. Tras el proceso diferenciador in vitro, las células mesenquimales
troncales humanas no expresan marcadores HLA (I y II) importantes en la respuesta inmune, pero sí expresan
débilmente proteínas relacionadas con los principales estratos epiteliales (CK5, CK6 y CK14).
Conclusión: La ausencia de expresión de marcadores de HLA I y II por citometría de flujo en las células diferenciadas
favorecería su uso con carácter alogénico en la construcción de piel y córneas humanas por ingeniería de
tejidos, sin embargo, son necesarios más estudios que confirmen estos resultados preliminares y protocolos que
optimicen el proceso diferenciador in vitro de las células mesenquimales troncales.Background: Human mesenchymal stem cells derived from adipose tissue and bone marrow are one of the most
common cell therapy procedures used in recent clinical trials due to their immunomodulation capacity. Furthermore,
for their epithelial differentiation potential can be used in tissue engineering, incorporated in artificial
tissues such as skin and cornea, replacing autologous epithelial cells. It is necessary to make a correct cellular
characterization of differentiated cells and to study the effect in HLA I and II expression.
Objetives: Characterization and quality controls under GMP conditions of in vitro differentiated human mesenchymal
stem cells from different sources (adipose tissue and bone marrow) to epithelial lineage, and study of HLA I
and II expression before and after differentiation.
Methods: Isolation and expansion of two human mesenchymal stem cells lines from their tissues of origin and in
vitro differentiation to epithelial cells using culture mediums supplemented with specific growth factors. Quality
controls according Good Manufacturing Practices have been made and HLA I and II expression before and after
differentiation have been studied. Finally, characteristics of differentiated cells have been demonstrated by histological
and immunohistochemical analysis.
Results: Two human mesenchymal stem cells lines from adipose tissue and bone marrow have been isolated
complying with the proposed quality controls. After in vitro differentiation, human mesenchymal stem cells do
not express HLA (I and II) markers, which are important in immune response, but weakly express proteins related
to main epithelial layers of human skin (CK5, CK6 and CK14).
Conclusion: The absence of expression of HLA I and II by flow cytometry in differentiated cells would promote the
use of them with allogenic character to construct human skin and cornea by tissue engineering, however, more
studies and protocols are required to confirm these preliminary results and to optimize in vitro differentiation of
human mesenchymal stem cells.FIS ISC-III and FEDER PI13/0257
A machine-learned analysis of human gene polymorphisms modulating persisting pain points to major roles of neuroimmune processes
Background Human genetic research has implicated functional variants of more than one hundred genes in the modulation of persisting pain. Artificial intelligence and machine-learning techniques may combine this knowledge with results of genetic research gathered in any context, which permits the identification of the key biological processes involved in chronic sensitization to pain. MethodsResultsBased on published evidence, a set of 110 genes carrying variants reported to be associated with modulation of the clinical phenotype of persisting pain in eight different clinical settings was submitted to unsupervised machine-learning aimed at functional clustering. Subsequently, a mathematically supported subset of genes, comprising those most consistently involved in persisting pain, was analysed by means of computational functional genomics in the Gene Ontology knowledgebase. Clustering of genes with evidence for a modulation of persisting pain elucidated a functionally heterogeneous set. The situation cleared when the focus was narrowed to a genetic modulation consistently observed throughout several clinical settings. On this basis, two groups of biological processes, the immune system and nitric oxide signalling, emerged as major players in sensitization to persisting pain, which is biologically highly plausible and in agreement with other lines of pain research. ConclusionsSignificanceThe present computational functional genomics-based approach provided a computational systems-biology perspective on chronic sensitization to pain. Human genetic control of persisting pain points to the immune system as a source of potential future targets for drugs directed against persisting pain. Contemporary machine-learned methods provide innovative approaches to knowledge discovery from previous evidence. We show that knowledge discovery in genetic databases and contemporary machine-learned techniques can identify relevant biological processes involved in Persitent pain.Peer reviewe
Genetic polymorphisms in folate pathway enzymes, DRD4 and GSTM1 are related to temporomandibular disorder
Journal Article; Research Support, Non-U.S. Gov't;BACKGROUND
Temporomandibular disorder (TMD) is a multifactorial syndrome related to a critical period of human life. TMD has been associated with psychological dysfunctions, oxidative state and sexual dimorphism with coincidental occurrence along the pubertal development. In this work we study the association between TMD and genetic polymorphisms of folate metabolism, neurotransmission, oxidative and hormonal metabolism. Folate metabolism, which depends on genes variations and diet, is directly involved in genetic and epigenetic variations that can influence the changes of last growing period of development in human and the appearance of the TMD.
METHODS
A case-control study was designed to evaluate the impact of genetic polymorphisms above described on TMD. A total of 229 individuals (69% women) were included at the study; 86 were patients with TMD and 143 were healthy control subjects. Subjects underwent to a clinical examination following the guidelines by the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Genotyping of 20 Single Nucleotide Polymorphisms (SNPs), divided in two groups, was performed by multiplex minisequencing preceded by multiplex PCR. Other seven genetic polymorphisms different from SNPs (deletions, insertions, tandem repeat, null genotype) were achieved by a multiplex-PCR. A chi-square test was performed to determine the differences in genotype and allelic frequencies between TMD patients and healthy subjects. To estimate TMD risk, in those polymorphisms that shown significant differences, odds ratio (OR) with a 95% of confidence interval were calculated.
RESULTS
Six of the polymorphisms showed statistical associations with TMD. Four of them are related to enzymes of folates metabolism: Allele G of Serine Hydoxymethyltransferase 1 (SHMT1) rs1979277 (OR = 3.99; 95%CI 1.72, 9.25; p = 0.002), allele G of SHMT1 rs638416 (OR = 2.80; 95%CI 1.51, 5.21; p = 0.013), allele T of Methylentetrahydrofolate Dehydrogenase (MTHFD) rs2236225 (OR = 3.09; 95%CI 1.27, 7.50; p = 0.016) and allele A of Methionine Synthase Reductase (MTRR) rs1801394 (OR = 2.35; 95CI 1.10, 5.00; p = 0.037). An inflammatory oxidative stress enzyme, Gluthatione S-Tranferase Mu-1(GSTM1), null allele (OR = 2.21; 95%CI 1.24, 4.36; p = 0.030) and a neurotransmission receptor, Dopamine Receptor D4 (DRD4), long allele of 48 bp-repeat (OR = 3.62; 95%CI 0.76, 17.26; p = 0.161).
CONCLUSIONS
Some genetic polymorphisms related to folates metabolism, inflammatory oxidative stress, and neurotransmission responses to pain, has been significantly associated to TMD syndrome.This study was supported by the Ministerio de Educación y Ciencia (Spain), (SAF2008-03314) and “Programa Torres Quevedo” (PTQ-09-0100496) GrantsYe