RESUMEN
Heterogeneity of Mesenchymal stem cells (MSCs) imposes limitations for their in vitro expansion and accounts for the lack of reproducibility in some clinical studies. So, this study was designed to isolate and enrich clones of multipotent and self-renewing MSCs from cord blood (CB). Enriched clones with higher proliferation and differentiation potential provide regenerative cells suitable for various clinical demands. MSCA and MSCB original (progenitor) cells were isolated from CB samples, and single cells were cloned by limiting dilution method, in mouse embryonic fibroblast conditioned media. Original MSCs and their single-cell derived clones were characterized by identifying their proliferation rate, immunophenotyping of surface antigens, expression of pluripotency and proliferation genes (Oct4, Sox2, Nanog, KLF4, c-Myc, and PDGFRA), and differentiation potential into multiple lineages (osteogenic, adipogenic, and chondrogenic). Some single-cell clones of MSCA showed a higher proliferation rate and greater differentiation potential than their original cells. However, original MSCB cells were of greater proliferation and differentiation potential than their derived single-cell clones, except for one clone which had comparable results. Cloning of MSCs was attainable when cultured in mouse embryonic fibroblast conditioned media. Single clones with higher proliferation and differentiation potential than their original progenitor cells were obtained by cloning of poorly functioning MSCs progenitor cells, enabling the selection of more therapeutically efficacious MSCs with better performance in clinical applications. Moreover, this study draws attention to the importance of CD105 as a possible MSCs biomarker associated with the multilineage commitment of MSCs.
Asunto(s)
Clonación Molecular/métodos , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/metabolismo , Adipogénesis/fisiología , Diferenciación Celular/fisiología , Proliferación Celular/fisiología , Células Cultivadas , Condrogénesis/fisiología , Sangre Fetal/citología , Humanos , Inmunofenotipificación , Factor 4 Similar a Kruppel , Reproducibilidad de los Resultados , Cordón Umbilical/citologíaRESUMEN
In Egypt, liver diseases are exceptionally high, maintaining the highest prevalence of hepatitis C virus (HCV) worldwide, and increasing rates of hepatocellular carcinoma (HCC). Available diagnostic methods show poor performance in early diagnosis of HCC. Definite pathogenic factors contributing in the development of HCV are still lacking. MicroRNAs have been reported as promising biomarkers for cancers diagnosis and in virus-host interaction. This study was conducted to detect the role of miR-182 and miR-150 as biomarkers for development of cirrhosis and malignant transformation in HCV infected patients. The expression of miR-182 and miR-150 was evaluated using real-time quantitative PCR (qRT-PCR) in 120 subjects: 40 HCC patients, 40 hepatitis C patients (20 cirrhotic and 20 non-cirrhotic HCV genotype 4) and 40 healthy controls. In HCC, statistically significant decrease of miR-182 and miR-150 compared to non-cirrhotic HCV patients (pâ¯=â¯0.015, pâ¯=â¯0.006 respectively) and of miR-150 compared to controls (pâ¯=â¯0.039). In cirrhotic HCV patients, significant down regulation of miR-182 and miR-150 compared to non-cirrhotic HCV (pâ¯=â¯0.003, pâ¯=â¯0.024 respectively). On the other hand, significant upregulation of miR-182 was observed in non-cirrhotic HCV compared to controls (pâ¯=â¯0.036). Alpha-fetoprotein (AFP) showed sensitivity 15% for HCC diagnosis at the cut-off value of 400â¯ng/ml, while combining AFP with miR-182 and miR-150, resulted in improving sensitivity to (90%) and diagnostic accuracy to (80%). miR-182 and miR-150 can be used as non invasive biomarkers for HCC and combination of these miRNAs and AFP markedly improve the diagnosis of HCC. Both miR-182 and miR-150 can also be used as predictive markers for detection of cirrhosis progression in HCV infected patients.
Asunto(s)
Carcinoma Hepatocelular/sangre , Hepatitis C Crónica/sangre , Cirrosis Hepática/sangre , Neoplasias Hepáticas/sangre , MicroARNs/sangre , Adulto , Anciano , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/genética , Carcinoma Hepatocelular/complicaciones , Carcinoma Hepatocelular/epidemiología , Carcinoma Hepatocelular/genética , Progresión de la Enfermedad , Egipto/epidemiología , Femenino , Regulación Neoplásica de la Expresión Génica , Hepatitis C Crónica/complicaciones , Hepatitis C Crónica/epidemiología , Hepatitis C Crónica/genética , Humanos , Cirrosis Hepática/complicaciones , Cirrosis Hepática/epidemiología , Cirrosis Hepática/genética , Neoplasias Hepáticas/complicaciones , Neoplasias Hepáticas/epidemiología , Neoplasias Hepáticas/genética , Masculino , MicroARNs/genética , Persona de Mediana Edad , Prevalencia , Curva ROC , Sensibilidad y Especificidad , alfa-Fetoproteínas/análisisRESUMEN
Egypt is confronted with the highest hepatitis C virus (HCV) epidemic. Apoptosis and cellular immune responses are crucial to the clearance or persistence of viral infections. This case-control study was carried out to detect whether apoptosis genes single nucleotide polymorphisms (SNPs) confer risk to HCV in a cohort of Egyptian patients and to explore their association with viral load. One hundred and ninety six blood samples were withdrawn from 96 HCV patients and 100 controls. The Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) -1525G>A and FasL-844T>C SNPs were genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Hepatitis C viral load was measured using Real time PCR. Results Genotypes distributions of TRAIL -1525G>A and FasL-844 T>C polymorphisms in controls were in accordance with Hardy-Weinberg equilibrium (p>0.05). The study showed a statistically significant difference in the distribution of the TRAIL -1525G>A polymorphism genotypes and the FasL-844 T>C polymorphism genotypes between the HCV patients and the controls (p=0.001 and 0.02 respectively), with association of the -1525GA genotype and -844 TT genotype with increased risk of HCV infection (OR=2.68, 1.942 respectively, 95% CI=1.482-4.846, 1.1-3.43, respectively). No significant association was detected between TRAIL, FasL and the viral load. Our results suggest that the FasL -844T>C SNP is implicated in the susceptibility to HCV in Egyptian patients and firstly report the involvement of TRAIL gene polymorphism in the risk of the disease. Therefore we recommend national programs to delineate genetic factors that may put individuals at risk for contracting HCV.
Asunto(s)
Epidemias , Proteína Ligando Fas/genética , Predisposición Genética a la Enfermedad , Hepatitis C/genética , Polimorfismo de Nucleótido Simple , Ligando Inductor de Apoptosis Relacionado con TNF/genética , Carga Viral , Anciano , Estudios de Casos y Controles , Egipto/epidemiología , Femenino , Técnicas de Genotipaje , Hepatitis C/epidemiología , Hepatitis C/inmunología , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Medición de RiesgoRESUMEN
BACKGROUND: Neonatal diabetes mellitus (NDM) is a monogenic form of diabetes mellitus. Until now, patients in developing countries who had this condition had been misdiagnosed as having type 1 diabetes mellitus and accordingly directed to erroneous, ineffective, and costly therapeutic regimens. OBJECTIVE: To detect Egyptian patients who harbor pathological variant in the KCNJ11 gene, so that their treatment regimen can be modified as needed to increase its effectiveness. METHODS: We sequenced KCNJ11 in 17 ethnic Egyptian probands diagnosed with diabetes mellitus before age 2 years. RESULTS: A preliminary case individual harboring a KCNJ11 pathological variant (p.R201H) was identified. The patient was successfully shifted from insulin therapy to sulfonylurea. Four previously identified benign variants, namely, E23K, I337V, L270V, and A190A, were detected in this patient. CONCLUSION: Implementing the findings of this molecular analysis could have a major clinical and nationwide economic impact on world health, especially in developing countries.
Asunto(s)
Análisis Mutacional de ADN , Diabetes Mellitus , Canales de Potasio de Rectificación Interna/genética , Adolescente , Niño , Preescolar , Estudios Transversales , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/tratamiento farmacológico , Diabetes Mellitus/genética , Diabetes Mellitus Tipo 1 , Errores Diagnósticos , Egipto , Femenino , Humanos , Lactante , Masculino , Mutación , Estudios Prospectivos , Compuestos de Sulfonilurea/uso terapéuticoRESUMEN
At least 1 in 10 of the Egyptian population aged 15-59 is burdened with hepatitis C virus (HCV) infection, stamping Egypt the highest country harboring HCV worldwide. Considerable evidence supported the involvement of host genetic factors in the pathogenesis of HCV and the possibility of implementation in target therapies. ApoB gene polymorphisms are postulated to affect the susceptibility of HCV infection. Hence, we aimed to evaluate the relationship between ApoB-516C/T promoter gene polymorphism and HCV infection in a cohort of Egyptian patients and to explore whether higher levels of low-density lipoprotein (LDL) might compete with lipoviral particles (LVP) in the binding to LDL receptor (LDLR), thus escaping infection. Ninety-seven HCV patients and 96 matched controls were enrolled in this study. We genotyped ApoB-516C/T using PCR-RFLP method. ApoB concentrations were measured by immunoturbidimetric assay. The genotype and the allele frequencies of ApoB-516C/T promoter gene polymorphism in cases were statistically insignificant compared with healthy individuals (P = 0.109, 0.125, respectively). Sex stratification showed significantly lower counts of C/T genotype in female patients compared with female controls (P = 0.011, OR = 0.132, 95% CI = 0.026-0.657). Significantly higher levels of LDL and ApoB were detected in the control group (P < 0.001). This study shows that the ApoB-516C/T promoter gene polymorphism has no impact on the risk of HCV infection. However, the C/T genotype may be a protective factor for our female cohort. Further studies with larger samples are needed to verify this genetic gender diversity. Additionally, high levels of LDL and ApoB might prevent HCV infection.
Asunto(s)
Apolipoproteína B-100/sangre , Apolipoproteína B-100/genética , Predisposición Genética a la Enfermedad , Hepatitis C/genética , Polimorfismo de Nucleótido Simple , Regiones Promotoras Genéticas , Adolescente , Adulto , Estudios de Cohortes , Egipto , Femenino , Técnicas de Genotipaje , Humanos , Masculino , Persona de Mediana Edad , Nefelometría y Turbidimetría , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Medición de Riesgo , Factores Sexuales , Adulto JovenRESUMEN
BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune disease, with multiple genetic and environmental factors involved in its etiology. The toll-like receptor 9 (TLR9) gene has been reported to have important roles in the development and progression of SLE. In this case-control study, the effect of TLR9 polymorphism on susceptibility to SLE was investigated in Egyptian patients. METHODS: We studied the distribution of the TLR9 rs352139 (G + 1174A) single nucleotide polymorphism (SNP) by allele-specific polymerase chain reaction (PCR) in 104 Egyptian patients with SLE and 108 age-, sex-, and ethnically matched controls. RESULTS: There was no statistically significant difference in the distribution of the AA genotype and alleles between SLE patients and the control group in our study; however, the GA heterozygous patients were three times more likely to develop SLE (P < 0.001). A significant association was detected between TLR9 genotypes and some of the disease manifestations as myositis (p = 0.032), psychosis (p = 0.014), photosensitivity (p = 0.002), and pleurisy (p = <0.001). Moreover, we observed a significant association between the TLR9 AA and GA genotypes and the presence of antinuclear antibodies (ANA) (p = 0.038). CONCLUSION: The G + 1174A SNP in the toll receptor 9 gene may contribute to the genetic susceptibility of SLE in Egyptian patients. Also, an influence for this polymorphism on disease manifestations has been elucidated.
Asunto(s)
Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Lupus Eritematoso Sistémico/genética , Polimorfismo de Nucleótido Simple , Receptor Toll-Like 9/genética , Adulto , Alelos , Estudios de Casos y Controles , Egipto , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Lupus Eritematoso Sistémico/diagnóstico , Oportunidad Relativa , Fenotipo , Adulto JovenRESUMEN
Fas/Fas ligand (FasL) system is the most critical apoptotic signaling entity in the extrinsic apoptotic pathway; hence mutations affecting this pathway may prevent the immune system from the removal of newly-formed tumor cells, and thus lead to tumor formation. The present study investigated the association between the FasL -844T/C polymorphism and the risk of hepatocellular carcinoma (HCC) in a cohort of Egyptian patients and explored the relationship of various clinical and pathological parameters with this single nucleotide polymorphism (SNP). Blood samples were withdrawn from hundred HCC patients and 100 age-, sex- and ethnically matched controls. The FasL -844T/C (rs763110) gene polymorphism was typed from genomic DNA using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay. Genotype distributions and allelic frequencies between patients and control subjects showed that the TT homozygous patients were two times more likely to develop HCC (p=0.011). Also, the T allele was found to be a significant risk factor for the disease (OR 1.970, 95% CI 1.250-3.105, p=0.003). No association was detected between different parameters of the disease and the SNP. For the first time, our results suggest that the -844T/C polymorphism in the FasL gene confers risk to HCC. The alarming increase in the incidence of HCC in Egypt encourages further studies to document our results in a larger sample, and recommends more genetic studies hoping to define a genomic risk prediction specific to this cancer in our population.
Asunto(s)
Carcinoma Hepatocelular/genética , Proteína Ligando Fas/genética , Neoplasias Hepáticas/genética , Anciano , Egipto , Proteína Ligando Fas/metabolismo , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , RiesgoRESUMEN
There is evidence supporting an association between cardiovascular disease (CVD) and periodontitis. We determined whether patients with chronic periodontitis, who are otherwise healthy individuals, have higher serum concentrations of emerging risk markers of CVD such as C-reactive protein (CRP) and interleukin 6 (IL-6) and investigated the effect of subsequent periodontal treatment on the levels of these markers. A total of 40 individuals were included in the study. Serum levels of CRP and IL-6 were estimated twice, once on the initial visits and the other 3 months after periodontal therapy. The mean CRP and IL-6 levels were significantly higher (P < .001) in the patients compared with controls and significantly decreased (P < .001) following periodontal treatment. This study suggests that periodontitis is a potential modifiable risk factor for CVD.