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Aterosclerosis , Medicamentos Herbarios Chinos , Hiperlipidemias , Hipertensión , Pueraria , Salvia miltiorrhiza , Amlodipino , Aterosclerosis/tratamiento farmacológico , Atorvastatina , Humanos , Hidroclorotiazida , Hiperlipidemias/tratamiento farmacológico , Hipertensión/tratamiento farmacológicoRESUMEN
SETTING: Shanghai is a mega city where 39% of the population comprises internal migrants. OBJECTIVE: To examine the different roles played by migrants and permanent residents in the transmission of multidrug-resistant tuberculosis (MDR-TB). DESIGN: We conducted a population-based cohort study to assess MDR-TB transmission in Shanghai between 1 January 2009 and 31 December 2012 using genotyping and geospatial analyses. RESULTS: A total of 367 MDR-TB cases formed the study cohort. Significant differences between MDR-TB cases who were internal migrants and those who were permanent residents were found with regard to age, sex, region, genetic characteristics and treatment outcomes. Permanent residents had a higher transmission rate than internal migrants (OR 3.36, 95%CI 1.86-6.09). Permanent residents and genotypic clustering cases had similar clusters in central downtown and some parts of suburban areas. Most of the clusters of internal migrants were found in rural areas bordering suburban areas. Clusters of genotypic non-clustering cases showed patterns that closely matched those of internal migrants, suggesting acquired drug resistance in migrants. CONCLUSION: In Shanghai, permanent residents were significantly associated with recent transmission of MDR-TB in central downtown areas. Clustered cases of internal migrants in rural areas were most likely to have contracted MDR-TB through acquired resistance.
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Emigración e Inmigración , Mycobacterium tuberculosis/genética , Migrantes , Tuberculosis Resistente a Múltiples Medicamentos/transmisión , Adolescente , Adulto , Anciano , Antituberculosos/uso terapéutico , China/epidemiología , Análisis por Conglomerados , Estudios de Cohortes , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Factores de Riesgo , Tuberculosis Resistente a Múltiples Medicamentos/tratamiento farmacológico , Adulto JovenRESUMEN
Dyslexia is a specific impairment in learning to read and has strong heritability. An intronic deletion within the DCDC2 gene, with ~8% frequency in European populations, is increasingly used as a marker for dyslexia in neuroimaging and behavioral studies. At a mechanistic level, this deletion has been proposed to influence sensory processing capacity, and in particular sensitivity to visual coherent motion. Our re-assessment of the literature, however, did not reveal strong support for a role of this specific deletion in dyslexia. We also analyzed data from five distinct cohorts, enriched for individuals with dyslexia, and did not identify any signal indicative of associations for the DCDC2 deletion with reading-related measures, including in a combined sample analysis (N=526). We believe we conducted the first replication analysis for a proposed deletion effect on visual motion perception and found no association (N=445 siblings). We also report that the DCDC2 deletion has a frequency of 37.6% in a cohort representative of the general population recruited in Hong Kong (N=220). This figure, together with a lack of association between the deletion and reading abilities in this cohort, indicates the low likelihood of a direct deletion effect on reading skills. Therefore, on the basis of multiple strands of evidence, we conclude that the DCDC2 deletion is not a strong risk factor for dyslexia. Our analyses and literature re-evaluation are important for interpreting current developments within multidisciplinary studies of dyslexia and, more generally, contribute to current discussions about the importance of reproducibility in science.
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Dislexia/genética , Proteínas Asociadas a Microtúbulos/genética , Adolescente , Adulto , Niño , Femenino , Eliminación de Gen , Predisposición Genética a la Enfermedad , Humanos , Masculino , Percepción de Movimiento , Factores de Riesgo , Adulto JovenAsunto(s)
Infecciones por Adenovirus Humanos/complicaciones , Adenovirus Humanos/aislamiento & purificación , Diabetes Mellitus/virología , Dislipidemias/virología , Heces/virología , Obesidad/virología , Adiposidad , Adulto , Anticuerpos Antivirales/sangre , Pueblo Asiatico , ADN Viral/sangre , Femenino , Hong Kong , Humanos , Masculino , Persona de Mediana Edad , Pruebas de NeutralizaciónRESUMEN
Metabolic disorders including type 2 diabetes, obesity and hypertension have growing prevalence globally every year. Genome-wide association studies have successfully identified many genetic markers associated to these diseases, but few studied their interaction effects. In this study, twenty candidate SNPs from sixteen genes are selected, and a lasso-multiple regression approach is implemented to consider the SNP-SNP interactions among them in an Asian population. It is found out that the main effects of the markers are weak but the interactions among the candidates showed a significant association to diseases. SNPs from genes CDKN2BAS and KCNJ11 are significantly associated to risk for developing diabetes, and SNPs from FTO and APOA5 might interact to play an important role for the onset of hypertension.
RESUMEN
Developmental dyslexia, the most common childhood learning disorder, is highly heritable, and recent studies have identified KIAA0319-Like (KIAA0319L) as a candidate dyslexia susceptibility gene at the 1p36-34 (DYX8) locus. In this experiment, we investigated the anatomical effects of knocking down this gene during rat corticogenesis. Cortical progenitor cells were transfected using in utero electroporation on embryonic day (E) 15.5 with plasmids encoding either: (1) Kiaa0319l small hairpin RNA (shRNA), (2) an expression construct for human KIAA0319L, (3) Kiaa0319l shRNA+KIAA0319L expression construct (rescue), or (4) controls (scrambled Kiaa0319l shRNA or empty expression vector). Mothers were injected with 5-bromo-2-deoxyuridine (BrdU) at either E13.5, E15.5, or E17.5. Disruption of Kiaa0319l function (by knockdown, overexpression, or rescue) resulted in the formation of large nodular periventricular heterotopia in approximately 25% of the rats, which can be seen as early as postnatal day 1. Only a small subset of heterotopic neurons had been transfected, indicating non-cell autonomous effects of the transfection. Most heterotopic neurons were generated in mid- to late-gestation, and laminar markers suggest that they were destined for upper cortical laminae. Finally, we found that transfected neurons in the cerebral cortex were located in their expected laminae. These results indicate that KIAA0319L is the fourth of four candidate dyslexia susceptibility genes that is involved in neuronal migration, which supports the association of abnormal neuronal migration with developmental dyslexia.
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Corteza Cerebral/crecimiento & desarrollo , Dislexia/genética , Regulación del Desarrollo de la Expresión Génica , Malformaciones del Desarrollo Cortical del Grupo II/genética , Células-Madre Neurales/metabolismo , Proteínas Nucleares/metabolismo , Animales , Animales Recién Nacidos , Susceptibilidad a Enfermedades , Electroporación , Humanos , Neurogénesis/genética , Proteínas Nucleares/genética , ARN Interferente Pequeño , Ratas , Ratas Transgénicas , Receptores de Superficie Celular , TransfecciónRESUMEN
AIMS: POU class 2 homeobox 1 (POU2F1), also known as octamer-binding transcription factor-1 (OCT-1), is a ubiquitous transcription factor that plays a key role in the regulation of genes related to inflammation and cell cycles. POU2F1 is located on chromosome 1q24, a region with linkage for Type 2 diabetes in Chinese and other populations. We examined the association of POU2F1 genetic variants with Type 2 diabetes in Hong Kong Chinese using two independent cohorts. METHODS: We genotyped five haplotype-tagging single nucleotide polymorphisms at POU2F1 in 1378 clinic-based patients with Type 2 diabetes and 601 control subjects, as well as 707 members from 179 families with diabetes. RESULTS: We found significant associations of rs4657652, rs7532692, rs10918682 and rs3767434 (OR = 1.26-1.59, 0.0003 < P(unadjusted) < 0.035) with Type 2 diabetes in the clinic-based case-control cohorts. Rs3767434 was also associated with Type 2 diabetes (OR = 1.55, P(unadjusted) = 0.013) in the family-based cohort. Meta-analysis revealed similar associations. In addition, the risk G allele of rs10918682 showed increased usage of insulin treatment during a mean follow-up period of 7 years [hazard ratio = 1.50 (1.05-2.14), P = 0.025]. CONCLUSIONS: Using separate cohorts, we observed consistent results showing the contribution of multiple variants at POU2F1 to the risk of Type 2 diabetes.
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Pueblo Asiatico/genética , Diabetes Mellitus Tipo 2/genética , Predisposición Genética a la Enfermedad , Factor 1 de Transcripción de Unión a Octámeros/genética , Polimorfismo de Nucleótido Simple , Adulto , Diabetes Mellitus Tipo 2/epidemiología , Femenino , Ligamiento Genético/genética , Genotipo , Hong Kong/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Filogenia , Factores de Riesgo , Factores de Transcripción/genéticaRESUMEN
OBJECTIVE: Our study aimed to assess the effectiveness of esomeprazole or rabeprazole in combination with amoxicillin and clarithromycin for the eradication of Helicobacter pylori in Hong Kong non-ulcer dyspepsia (NUD) patients. METHODS: A prospective clinical trial was conducted at the Alice Ho Miu ling Nethersole Hospital outpatient endoscopy center from June 2004 to December 2005. Participants received amoxicillin 1 g, clarithromycin 500 mg, and, esomeprazole 20 mg (EAC) or rabeprazole 20 mg (RAC), all given twice daily for 1 week. The H. pylori status was determined by the [13C] urea breath test at least 4 weeks after completion of the treatment. Mutation status of CYP2C19 in exon 4 and exon 5 associated with the poor metabolizer phenotype was determined. RESULTS: The intention-to-treat eradication rates in patients treated with RAC and EAC were 77% and 84.6% respectively, and per protocol-based eradication rates were 83.7% and 88.9% respectively. The eradication rates did not vary with CYP2C19 phenotype found. For clarithromycin-sensitive strains, the cure rates were statistically significant regardless of CYP2C19 polymorphism (P < 0.0001). CONCLUSION: Triple therapy with either EAC or RAC is effective for Hong Kong Chinese NUD patients with H. pylori infection. Success eradication was related to clarithromycin resistance and not CYP2C19 genotype.
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2-Piridinilmetilsulfinilbencimidazoles/uso terapéutico , Antibacterianos/uso terapéutico , Esomeprazol/uso terapéutico , Infecciones por Helicobacter/tratamiento farmacológico , 2-Piridinilmetilsulfinilbencimidazoles/administración & dosificación , Adulto , Amoxicilina/administración & dosificación , Amoxicilina/uso terapéutico , Antibacterianos/administración & dosificación , Antiulcerosos/administración & dosificación , Antiulcerosos/uso terapéutico , Hidrocarburo de Aril Hidroxilasas/genética , Pueblo Asiatico/genética , Pruebas Respiratorias/métodos , Claritromicina/administración & dosificación , Claritromicina/uso terapéutico , Citocromo P-450 CYP2C19 , Farmacorresistencia Bacteriana , Dispepsia/tratamiento farmacológico , Dispepsia/microbiología , Esomeprazol/administración & dosificación , Femenino , Infecciones por Helicobacter/microbiología , Helicobacter pylori/efectos de los fármacos , Helicobacter pylori/aislamiento & purificación , Hong Kong , Humanos , Masculino , Persona de Mediana Edad , Mutación , Farmacogenética , Polimorfismo Genético , Estudios Prospectivos , Rabeprazol , Urea/metabolismoRESUMEN
OBJECTIVE: Childhood obesity is a growing global epidemic. Recent studies indicate that obesity and related metabolic traits are highly heritable. Increasing evidence suggests that growth hormone (GH) and the insulin-like growth factor-I (IGF-I) axis have important functions in regulating adiposity and insulin sensitivity. Five single-nucleotide polymorphisms (SNPs) at IGF-binding protein-3 (IGFBP3) were genotyped to find their associations with IGF-1 activity level and common clinical metabolic traits. PATIENTS AND METHODS: We examined the associations of five SNPs at IGFBP3 with serum IGF-I and IGFBP-3 levels, as well as with obesity-related metabolic traits in 981 Hong Kong Chinese adolescents. Factor analysis was used to reduce the intercorrelated variables to five factor scores indicating body composition, blood pressure, IGF-I activity, triglyceride (TG)+high-density lipoprotein cholesterol (HDL-C) and total cholesterol (TC)+low-density lipoprotein cholesterol (LDL-C) factor scores. RESULTS: There was a strong association between the -202A/C polymorphism (rs2854744) and IGF-I activity (P=1.2 x 10(-6)) and TC+LDL-C factor scores (P=0.0085), corrected for age and sex. The C allele was associated with decreased IGFBP-3 levels (P=1.21 x 10(-13)), increased IGF-I/IGFBP-3 molar ratio (P=5.22 x 10(-6)) and decreased LDL-C (P=0.020). There was also a significant association between a G/A polymorphism at the 3' flanking sequence (rs13223993) of the IGFBP3 gene and the TG+HDL-C factor score (P=0.0013). The minor A allele carriers of rs13223993 had a lower HDL-C (P=0.0067) level and a tendency toward a high TG level. Haplotype analysis did not increase the significance of associations between single SNPs and phenotypes. CONCLUSION: Our results support the function of IGFBP3 gene polymorphisms in modulating IGF-I activity and lipid levels in adolescents. Given the prognostic significance of IGF-I, IGFBPs and lipids on risk of diabetes, obesity and cancer, long-term studies are required to clarify the clinical meaning of these findings.
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Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Lípidos/sangre , Obesidad/sangre , Adolescente , Composición Corporal/genética , Femenino , Genotipo , Hong Kong/epidemiología , Humanos , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/sangre , Lípidos/genética , Masculino , Obesidad/epidemiología , Fenotipo , Polimorfismo de Nucleótido Simple/genéticaRESUMEN
BACKGROUND: Single-nucleotide polymorphism (SNP)-based genome-wide association study revealed that markers on chromosome 17q21 were linked to childhood asthma but not atopy in Caucasians, with the strongest signal being detected for the SNP rs7216389 in the ORMDL3 gene. Such association was unknown in Chinese. This study delineated the allele and genotype frequencies of 10 SNPs at chromosome 17q21, and investigated the relationship between these SNPs and asthma and plasma IgE in southern Chinese children. METHODS: Asthmatic children and non-allergic controls were recruited from pediatric clinics. Their plasma total and aeroallergen-specific IgE concentrations were measured by immunoassay. Ten SNPs on 17q21 region were genotyped by multiplex SNaPshot, and their genotype associations with asthma traits analyzed using multivariate regression. RESULTS: 315 patients and 192 controls were enrolled. The allele frequency for C allele of rs7216389 varied significantly from 0.232 in our controls, 0.389 in Han Chinese to 0.536 in Caucasians. Asthma diagnosis was associated with rs11650680 and five other SNPs including rs7216389 (P = 0.019-0.034), whereas atopy was associated only with rs11650680 (P = 0.0004). Linear regression revealed the covariates for plasma total IgE to be significant for rs11650680 (P = 0.008-0.0002). Haplotypic associations were found with atopy and increased plasma total IgE, with the respective odds ratios and 95% confidence intervals for TTTCCGTT haplotype to be 0.21 and 0.09-0.52 (P = 0.0002) and 0.41 and 0.18-0.90 (P = 0.025). CONCLUSION: Childhood asthma and atopy are associated with chromosome 17q21 in Chinese, but such association may involve genes other than ORMDL3 in this region.
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Asma/genética , Cromosomas Humanos Par 17/genética , Predisposición Genética a la Enfermedad , Hipersensibilidad Inmediata/genética , Adolescente , Pueblo Asiatico , Estudios de Casos y Controles , Niño , Preescolar , Marcadores Genéticos , Estudio de Asociación del Genoma Completo , Humanos , Inmunoglobulina E/sangre , Proteínas de la Membrana/genética , Polimorfismo de Nucleótido SimpleRESUMEN
Traditional Chinese medicine (TCM) has been used for prevention and treatment of severe acute respiratory syndrome (SARS) in Hong Kong during the outbreak in spring 2003. We investigated the immunomodulating effects of an innovative TCM regimen derived from two herbal formulas (Sang Ju Yin and Yu Ping Feng San) for treating febrile diseases. Thirty-seven healthy volunteers were given the oral TCM regimen daily for 14 days. Peripheral venous blood samples were taken on days 0, 15 and 29 for hematology, biochemistry and immunology tests, including the measurement of blood lymphocyte subsets and plasma T-helper lymphocyte types 1 and 2 cytokines and receptor. After 3 months, 23 of the volunteers participated in a control study without TCM treatment for the same time course of blood tests. Two volunteers withdrew on day 2, due to headache and dizziness. All others remained well without any side effects. No participants showed significant changes in their blood test results, except that the T-lymphocyte CD4/CD8 ratio increased significantly from 1.31 +/- 0.50 (mean +/- SD) on day 0 to 1.41 +/- 0.63 on day 15 (p < 0.02), and reduced to 1.32 +/- 0.47 on day 29 (p < 0.05). In the control study, there were no changes in the CD4/CD8 ratio. The transient increase in CD4/CD8 ratio was likely due to the TCM intake. We postulate that the administration of the innovative TCM may have beneficial immunomodulatory effects for preventing viral infections including SARS.
Asunto(s)
Relación CD4-CD8 , Medicamentos Herbarios Chinos/uso terapéutico , Medicina Tradicional China , Adulto , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Hepatitis B virus (HBV)-encoded X antigen (HBxAg) contributes to the development of hepatocellular carcinoma (HCC). A frequent characteristic of HCC is reduced or absent expression of the cell adhesion protein, E-cadherin, although it is not known whether HBxAg plays a role. To address this, the levels of E-cadherin were determined in HBxAg-positive and -negative HepG2 cells in culture, and in tumor and surrounding nontumor liver from a panel of HBV carriers. The results showed an inverse relationship between HBxAg and E-cadherin expression both in tissue culture and in vivo. In HBxAg-positive cells, E-cadherin was suppressed at both the mRNA and protein levels. This was associated with hypermethylation of the E-cadherin promoter. Depressed E-cadherin correlated with HBxAg trans-activation function, as did the migration of HepG2 cells in vitro. Decreased expression of E-cadherin was also associated with the accumulation of beta-catenin in the cytoplasm and/or nuclei in tissues and cell lines, which is characteristic of activated beta-catenin. Additional work showed that HBxAg-activated beta-catenin. Together, these results suggest that the HBxAg is associated with decreased expression of E-cadherin, accumulation of beta-catenin in the cytoplasm and nucleus, and increased cell migration, which may contribute importantly to hepatocarcinogenesis.
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Cadherinas/metabolismo , Carcinoma Hepatocelular/virología , Neoplasias Hepáticas/virología , Transactivadores/metabolismo , Cadherinas/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Movimiento Celular , Núcleo Celular/química , Citoplasma/química , Metilación de ADN , Regulación hacia Abajo , Virus de la Hepatitis B , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Regiones Promotoras Genéticas , Transporte de Proteínas , Transactivadores/análisis , Transactivadores/genética , Transfección , Proteínas Reguladoras y Accesorias Virales , beta Catenina/análisis , beta Catenina/metabolismoRESUMEN
Severe acute respiratory syndrome (SARS) is a global health concern. In Hong Kong, two major outbreaks, one hospital based and the other in the Amoy Gardens apartments, were identified. The frequency of diarrhoea, admission to intensive care, and mortality differed significantly between the two outbreaks. We did genomic sequencing for viral isolates from five Amoy Gardens patients. The virus sequence was identical in four of these five patients. The sequence data from one hospital case and the four identical community cases had only three nucleotide differences. Alterations in the SARS coronavirus genome are unlikely to have caused the distinctive clinical features of the Amoy Gardens patients, and these results highlight the importance of non-viral genomic factors in this outbreak.
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Brotes de Enfermedades/estadística & datos numéricos , Genoma Viral , Síndrome Respiratorio Agudo Grave/virología , Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo/genética , Secuencia de Bases/genética , Infección Hospitalaria/virología , Hong Kong/epidemiología , Humanos , Síndrome Respiratorio Agudo Grave/diagnóstico , Síndrome Respiratorio Agudo Grave/epidemiologíaRESUMEN
Elfin (previously named CLIM1) is a protein that possesses an N-terminal PDZ domain and a C-terminal LIM domain. It belongs to the family of Enigma proteins. Enigma proteins are a family of cytoplasmic proteins that contain an N-terminal PDZ domain and a series of C-terminal LIM domains. By virtue of these two protein interacting domains, Enigma proteins are capable of protein-protein interactions. It has been proposed that Enigma proteins may act as adapters between kinases and the cytoskeleton. We have previously shown that Elfin is most abundantly expressed in the heart and it colocalizes with alpha-actinin 2 at the Z-disks of the myocardium. In this report, Elfin was shown to localize at the actin stress fibers of myoblasts, as revealed by green fluorescent protein (GFP) tagging. In situ hybridization and immunostaining showed that Elfin expression begins at an early stage in mouse development and is present throughout the developing heart. Taken together, our experimental results suggest that Elfin may play an important role in myofibrillogenesis and heart development.
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Regulación del Desarrollo de la Expresión Génica , Corazón/embriología , Miocardio/metabolismo , Factores de Transcripción/biosíntesis , Factores de Transcripción/química , Actinina/metabolismo , Secuencia de Aminoácidos , Animales , Línea Celular , Citoplasma/metabolismo , Citoesqueleto/metabolismo , ADN Complementario/metabolismo , Proteínas Fluorescentes Verdes , Hibridación in Situ , Proteínas con Dominio LIM , Proteínas Luminiscentes/metabolismo , Ratones , Datos de Secuencia Molecular , Unión Proteica , Estructura Terciaria de Proteína , ARN Mensajero/metabolismo , Homología de Secuencia de Aminoácido , Fibras de Estrés/metabolismo , Factores de Tiempo , Distribución TisularRESUMEN
Yeast Apc11p together with Rbx1 and Roc2/SAG define a new class of RING-H2 fingers in a superfamily of E3 ubiquitin ligases. The human homolog of Apc11p, ANAPC11 was identified during a large-scale partial sequencing of a human liver cancer cDNA library and partial characterization was performed. This 514 bp full-length cDNA has a predicted open reading frame (ORF) encoding 84 amino acids. The ORF codes for ANAPC11, the human anaphase promoting complex subunit 11 (yeast APC11 homolog), which possesses a RING-H2 finger motif and exhibits sequence similarity to subunits of E3 ubiquitin ligase complexes. In Northern blot hybridization with poly(A) RNA of various human tissues using radio-labelled ANAPC11 cDNA probe, we found strong signals detected in skeletal muscle and heart; moderate signals detected in brain, kidney, and liver; and detectable but low signals in colon, thymus, spleen, small intestine, placenta, lung, and peripheral blood leukocyte. The ANAPC11 gene is located at the human chromosome 17q25. ANAPC11 is distributed diffusely in the cytoplasm and nucleus with discrete accumulation in granular structures in all the cell lines (AML 12, HepG2, and C2C12) transfected. Expression level of ANAPC11 is found higher in certain types of cancer determined in the RNA dot blot experiment.
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Carcinoma Hepatocelular/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Ligasas/genética , Neoplasias Hepáticas/metabolismo , Proteínas de Neoplasias , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Complejos de Ubiquitina-Proteína Ligasa , Ciclosoma-Complejo Promotor de la Anafase , Animales , Subunidad Apc11 del Ciclosoma-Complejo Promotor de la Anafase , Secuencia de Bases/genética , Biomarcadores de Tumor/metabolismo , Encéfalo/metabolismo , Carcinoma Hepatocelular/genética , Mapeo Cromosómico/métodos , Clonación Molecular , ADN Complementario/análisis , ADN de Neoplasias/análisis , Proteínas Fúngicas/genética , Humanos , Células Híbridas , Riñón/metabolismo , Leucemia/metabolismo , Hígado/metabolismo , Neoplasias Hepáticas/genética , Datos de Secuencia Molecular , Músculo Esquelético/metabolismo , Miocardio/metabolismo , Complejo Represivo Polycomb 1 , Fracciones Subcelulares/metabolismo , Distribución Tisular/fisiología , Células Tumorales Cultivadas/metabolismo , Ubiquitina-Proteína Ligasas , Levaduras/genéticaRESUMEN
We have cloned and characterized another alternatively spliced isoform of the human four-and-a-half LIM domain protein 1 (FHL1), designated FHL1C. FHL1C contains a single zinc finger and two tandem repeats of LIM domains at the N-terminus followed by a putative RBP-J binding region at the C-terminus. FHL1C shares the same N-terminal two-and-a-half LIM domains with FHL1 but different C-terminal protein sequences. Due to the absence of the exon 4 in FHL1C, there is a frame-shift in the 3' coding region. Sequence analysis indicated that FHL1C is the human homolog of murine KyoT2. The Northern blot and RT-PCR results revealed that FHL1 is widely expressed in human tissues, including skeletal muscle and heart at a high level, albeit as a relatively low abundance transcript in brain, placenta, lung, liver, kidney, pancreas, and testis. In contrast, FHL1C is specifically expressed in testis, skeletal muscle, and heart at a relatively low level compared with FHL1. The expression of FHL1C transcripts was also seen in aorta, left atrium, left, and right ventricles of human heart at low level. Immunoblot analysis using affinity-purified anti-FHL1C antipeptide antibodies confirmed a 20 kDa protein of FHL1C in human skeletal muscle and heart. Unlike FHL1B, which is another FHL1 isoform recently reported by our group and localized predominantly in the nucleus [Lee et al., 1999], FHL1C is localized both in the nucleus and cytoplasm of mammalian cell.
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Proteínas de Drosophila , Proteínas de Homeodominio/genética , Proteínas Musculares , Músculo Esquelético/metabolismo , Miocardio/metabolismo , Testículo/metabolismo , Empalme Alternativo , Animales , Secuencia de Bases/genética , Northern Blotting/métodos , Encéfalo/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Exones/genética , Mutación del Sistema de Lectura/genética , Proteínas de Homeodominio/química , Proteínas de Homeodominio/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular , Proteínas con Dominio LIM , Masculino , Ratones , Datos de Secuencia Molecular , Isoformas de Proteínas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/instrumentación , Homología de Secuencia , Fracciones Subcelulares/química , Fracciones Subcelulares/metabolismo , Secuencias Repetidas en Tándem/genética , Factores de Transcripción/química , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Dedos de Zinc/fisiologíaRESUMEN
Ilex latifolia is widely used as an ingredient to prepare traditional beverage drinks in southern China. In fact, various Ilex species have been used in Chinese folk medicine to treat coronary heart diseases. The mode of action is believed to be mediated by their coronary vasodilative effects. In this study, the water extract of the leaves of Ilex latifolia (IK-TP) was shown to increase the contractility and decrease the frequency of contraction in an isolated rat heart perfusion system. IK-TP was found to inhibit Na+/K+-ATPase activities in rat heart sarcolemma, rat brain microsomes and a purified enzyme from porcine cerebral cortex. IK-TP also inhibited Ca2+-dependent ATPase at a similar dose. Following exposure of the isolated rat heart to IK-TP at a dose that produces pronounced cardiac effects, inhibition of Na+/K+-ATPase activity can be readily detected in the heart. This study suggests the presence of ATPase inhibitory compounds in Ilex latifolia with specificities different from that of ouabain.
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Medicamentos Herbarios Chinos , Inhibidores Enzimáticos/farmacología , Frecuencia Cardíaca/efectos de los fármacos , Contracción Miocárdica/efectos de los fármacos , Extractos Vegetales/farmacología , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidores , Animales , Encéfalo/enzimología , Encéfalo/ultraestructura , ATPasas Transportadoras de Calcio/antagonistas & inhibidores , Cardiotónicos/farmacología , Corteza Cerebral/enzimología , Cinética , Masculino , Microsomas/enzimología , Miocardio/enzimología , Ratas , Ratas Sprague-Dawley , Sarcolema/enzimología , PorcinosRESUMEN
LIM domain proteins are found to be important regulators in cell growth, cell fate determination, cell differentiation and remodeling of the cell cytoskeleton. Human Four-and-a-half LIM-only protein 3 (FHL3) is a type of LIM-only protein that contains four tandemly repeated LIM motifs with an N-terminal single zinc finger (half LIM motif). FHL3 expresses predominantly in human skeletal muscle. In this report, FHL3 was shown to be a novel interacting partner of FHL2 using the yeast two-hybrid assay. Furthermore, site-directed mutagenesis of FHL3 indicated that the LIM2 of FHL3 is the essential LIM domain for interaction with FHL2. Green fluorescent protein (GFP) was used to tag FHL3 in order to study its distribution during myogenesis. Our result shows that FHL3 was localized in the focal adhesions and nucleus of the cells. FHL3 mainly stayed in the focal adhesion during myogenesis. Moreover, using site-directed mutagenesis, the LIM1 of FHL3 was identified as an essential LIM domain for its subcellular localization. Mutants of GFP have given rise to a novel technique, two-fusion fluorescence resonance energy transfer (FRET), in the determination of protein-protein interaction at particular subcellular locations of eukaryotic cells. To determine whether FHL2 and FHL3 can interact with one another and to locate the site of this interaction in a single intact mammalian cell, we fused FHL2 and FHL3 to different mutants of GFP and studied their interactions using FRET. BFP/GFP fusion constructs were cotransfected into muscle myoblast C2C12 to verify the colocalization and subcellular localization of FRET. We found that FHL2 and FHL3 were colocalized in the mitochondria of the C2C12 cells and FRET was observed by using an epi-fluorescent microscope equipped with an FRET specific filter set.
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Proteínas de Homeodominio/metabolismo , Proteínas Musculares , Factores de Transcripción , Secuencia de Bases , Línea Celular , Núcleo Celular/metabolismo , Cartilla de ADN , Proteínas Fluorescentes Verdes , Proteínas de Homeodominio/genética , Humanos , Péptidos y Proteínas de Señalización Intracelular , Proteínas con Dominio LIM , Proteínas con Homeodominio LIM , Proteínas Luminiscentes/metabolismo , Mutagénesis Sitio-Dirigida , Unión Proteica , Espectrometría de Fluorescencia , Técnicas del Sistema de Dos HíbridosRESUMEN
LIM domain proteins are found to be important regulators in cell growth, cell fate determination, cell differentiation, and remodeling of the cell cytoskeleton. Human Four-and-a-half LIM-only protein 2 (FHL2) is expressed predominantly in human heart and is only slightly expressed in skeletal muscle. Since FHL2 is an abundant protein in human heart, it may play an important role in the regulation of cell differentiation and myofibrillogenesis of heart at defined subcellular compartment. Therefore, we hypothesized that FHL2 act as a multi-functional protein by the specific arrangement of the LIM domains of FHL2 and that one of the LIM domains of FHL2 can function as an anchor and localizes it into a specific subcellular compartment in a cell type specific manner to regulate myofibrillogenesis. From our results, we observed that FHL2 is localized at the focal adhesions of the C2C12, H9C2 myoblast as well as a nonmyogenic cell line, HepG2 cells. Colocalization of vinculin-CFP and FHL2-GFP at focal adhesions was also observed in cell lines. Site-directed mutagenesis, in turn, suggested that the second LIM domain-LIM2 is essential for its specific localization to focal adhesions. Moreover, FHL2 was observed along with F-actin and focal adhesion of C2C12 and H9C2 myotubes. Finally, we believe that FHL2 moves from focal adhesions and then stays at the Z-discs of terminally differentiated heart muscle.
Asunto(s)
Proteínas del Ojo/metabolismo , Adhesiones Focales/metabolismo , Proteínas de Homeodominio/metabolismo , Proteínas Musculares , Músculos/ultraestructura , Miocardio/ultraestructura , Miofibrillas/metabolismo , Factores de Transcripción , Actinas/metabolismo , Secuencia de Aminoácidos , Diferenciación Celular , Línea Celular , Adhesiones Focales/ultraestructura , Proteínas Fluorescentes Verdes , Proteínas de Homeodominio/química , Humanos , Inmunohistoquímica , Proteínas con Homeodominio LIM , Proteínas Luminiscentes/metabolismo , Proteínas de la Membrana/metabolismo , Microscopía Confocal , Datos de Secuencia Molecular , Músculos/citología , Músculos/metabolismo , Mutagénesis Sitio-Dirigida , Miocardio/metabolismo , Estructura Terciaria de Proteína , Proteínas Recombinantes de Fusión/metabolismo , Alineación de Secuencia , Células Tumorales Cultivadas , Vinculina/metabolismoRESUMEN
BACKGROUND: The discovery of Cytochrome P450 2D6 (CYP 2D6) polymorphism is implicated in individual differences in drug metabolism rate. Mutation with defective alleles is associated with reduced metabolism of many anti-psychotic drugs metabolized by CYP 2D6. This may contribute to the development of tardive dyskinesia (TD) in patients with prolonged exposure to anti-psychotic drugs. METHODS: In this controlled study, the genotype of CYP 2D6*10 alleles, movement disorders and clinical characteristics in 38 Chinese schizophrenic patients with TD were compared with 38 age- and sex-matched schizophrenia patients without TD. RESULTS: There was no significant correlation between CYP 2D6*10 genotypes and TD in men. However, a significant increase in the frequency of CYP 2D6*10 allele was found in female patients with TD. CONCLUSIONS: The sex differences in CYP 2D6 genotyping and vulnerability to develop TD suggest that a biological predisposition that affects pharmacokinetics may be more significant in women, whereas other factors may be more important in men.