RESUMEN
The multimarker approach using Luminex technology represents a new tool for studying the pathogenesis of cardiovascular disease. Although many cardiac biomarkers in heart failure have been well established, the role and significance of their measurement in hypertensive patients is still questionable. The aim of our study was to evaluate the relationship of selected biomarkers in L-NAME-induced hypertension to the left ventricular remodeling in the two different periods of hypertension development. Four groups of 3-month-old male Wistar rats were investigated: (1) control 4 (placebo for 4 weeks), (2) control 7 (placebo for 7 weeks), (3) L-NAME 4 (40 mg/kg/day for 4 weeks), and (4) L-NAME 7 (40 mg/kg/day for 7 weeks). BNP, cTnI, TNF-α, and VEGF were measured using Rat CVD Panel 1 Kit (Milliplex® MAP). Cardiac troponin T was determined using Elecsys® Troponin T high sensitive immunoassay (Roche, Switzerland). Although the systolic blood pressure increases about 50% in L-NAME-induced hypertension in rat, both hypertrophy and fibrosis were expressed only slightly in this experiment. The levels of BNP, TNF-α, or VEGF did not differ significantly among groups. However, cardiac troponin T measured by high sensitive ELISA was significantly (P<0.05) increased in L-NAME 4 (0.229 µg/l versus 0.034 µg/l) and L-NAME-7 groups (0.366 µg/l versus 0.06 µg/l) in comparison with the controls. We conclude that the slightly increased cTnT levels could indicate ischemic damage of L-NAME-hypertensive heart. Importantly, to our best knowledge, this is the first study indicating that CVD rat panel may be a useful methodological tool in experimental cardiology.
Asunto(s)
Hipertensión/sangre , Animales , Biomarcadores/sangre , Presión Sanguínea/efectos de los fármacos , Inhibidores Enzimáticos , Hipertensión/inducido químicamente , Hipertensión/fisiopatología , Inmunoensayo , Masculino , NG-Nitroarginina Metil Éster , Péptido Natriurético Encefálico/sangre , Óxido Nítrico Sintasa/antagonistas & inhibidores , Ratas , Ratas Wistar , Troponina I/sangre , Troponina T/sangre , Factor de Necrosis Tumoral alfa/sangre , Factor A de Crecimiento Endotelial Vascular/sangreRESUMEN
Primary OA is a common multifactorial disease with not fully clarified molecular factors influencing the development of the disease. Among factors disturbing the cartilage integrity are cytokines, such as IL-1, which can stimulate proteinases, resulting in the cartilage destruction. In this regard, IL-1RA competing with IL-1 for binding to its receptor may act as an inhibitor of cartilage breakdown. Because of the possible functional implications, we tested VNTR polymorphism in the second intron of the IL-1RN gene as a putative factor of susceptibility to knee OA. Fifty patients with primary knee OA (diagnosed according to ACR criteria) and 170 healthy controls were included into the study. PCR using primers flanking the VNTR region containing variable numbers of an 86-bp tandem repeat was employed to test the hypothesis. An increased frequency and carriage rate of the IL-1RN*2 allele was found in OA patients in comparison with controls (28 % vs. 15 %, P = 0.0013, OR = 2.97; 95% CI 1.55-5.68 for frequency; 52.5 % vs. 25.3 %, P = 0.0019, OR = 2.95; 95% CI 1.54-5.68 for carriage rate). In addition, a higher frequency of genotype IL-1RN*1/*2 in OA patients was observed as compared with controls (42 % vs. 20.6 %, P = 0.0032, OR = 2.79; 95% CI 1.42-5.48). These results suggest that the IL-1RN*2 allele might represent a factor of susceptibility to OA; however, no correlation between this allele and the markers of cartilage degradation was found.
Asunto(s)
Antirreumáticos/metabolismo , Biomarcadores/metabolismo , Cartílago Articular/patología , Proteína Antagonista del Receptor de Interleucina 1 , Osteoartritis de la Rodilla/genética , Osteoartritis de la Rodilla/patología , República Checa , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Proteína Antagonista del Receptor de Interleucina 1/genética , Proteína Antagonista del Receptor de Interleucina 1/metabolismo , Repeticiones de Minisatélite , Osteoartritis de la Rodilla/inmunología , Polimorfismo Genético , Receptores de Interleucina-1/genética , Receptores de Interleucina-1/metabolismo , Población Blanca/genéticaRESUMEN
OBJECTIVE: The cartilage proteins melanoma inhibitory activity (MIA) and human cartilage gp-39 (HC gp-39) are candidate autoantigens in rheumatoid arthritis (RA). The present study was undertaken to investigate the endogenous HLA-DR4-restricted presentation of these self proteins, in order to seek in vivo evidence in support of their potential immunologic role. METHODS: MIA and HC gp-39 were assessed in synovial fluid (SF) by enzyme-linked immunosorbent assay and in synovial tissue (ST) by immunohistochemistry. Presentation by SF cells was investigated using specific, HLA-DR-restricted T cell hybridomas. RESULTS: MIA and HC gp-39 were detected in RA SF and ST, as well as in specimens from patients with other forms of arthritis. When HC gp-39-specific and MIA-specific HLA-DR4-restricted T cell hybridomas raised in HLA-DR4-transgenic mice were incubated with RA SF cells as antigen-presenting cells in the presence of HC gp-39 or MIA peptides, the corresponding T cell hybridomas showed strong responses, which were blocked by anti-HLA-DR antibodies. Weaker but qualitatively similar responses were observed with exogenous protein, indicating uptake and processing of these antigens by SF cells. More importantly, without addition of peptide or protein, endogenous presentation of MIA and HC gp-39 was detected in SF cells from 53% and 80% of HLA-DRB1*0401-positive RA patients, respectively. In addition, SF cells from 3 of 10 patients with spondylarthritis exhibited endogenous HC gp-39 presentation. CONCLUSION: These data indicate that immunodominant epitopes of MIA and HC gp-39 are actively presented in an HLA-DR-restricted manner in the inflamed RA joint. The question remains as to whether this leads to activation of autoreactive T cells, which could play a role in either the immunopathology or the immunomodulation of arthritis.
Asunto(s)
Artritis Reumatoide/inmunología , Proteínas de la Matriz Extracelular/análisis , Glicoproteínas/análisis , Antígenos HLA-DR/inmunología , Inflamación/inmunología , Articulaciones/inmunología , Proteínas de Neoplasias/análisis , Adipoquinas , Animales , Proteína 1 Similar a Quitinasa-3 , Ensayo de Inmunoadsorción Enzimática , Antígeno HLA-DR4/inmunología , Cadenas HLA-DRB1 , Prueba de Histocompatibilidad , Humanos , Hibridomas/inmunología , Articulaciones/fisiopatología , Lectinas , Ratones , Ratones Transgénicos , Líquido Sinovial/química , Linfocitos T/inmunologíaRESUMEN
Genetic variation in the interferon regulatory factor 5 (IRF5) gene affects systemic lupus erythematosus (SLE) susceptibility. However, association is complex and incompletely defined. We obtained fourteen European sample collections with a total of 1383 SLE patients and 1614 controls to better define the role of the different IRF5 variants. Eleven polymorphisms were studied, including nine tag single nucleotide polymorphisms (SNPs) and two extra functional polymorphisms. Two tag SNPs showed independent and opposed associations: susceptibility (rs10488631, P<10(-17)) and protection (rs729302, P<10(-6)). Haplotype analyses showed that the susceptibility haplotype, identified by the minor allele of rs10488631, can be due to epistasis between three IRF5 functional polymorphisms. These polymorphisms determine increased mRNA expression, a splice variant with a different exon 1 and a longer proline-rich region in exon 6. This result is striking as none of the three polymorphisms had an independent effect on their own. Protection was independent of these polymorphisms and seemed to reside in the 5' side of the gene. In conclusion, our results help to understand the role of the IRF5 locus in SLE susceptibility by clearly separating protection from susceptibility as caused by independent polymorphisms. In addition, we have found evidence for epistasis between known functional polymorphisms for the susceptibility effect.
Asunto(s)
Epistasis Genética , Predisposición Genética a la Enfermedad , Factores Reguladores del Interferón/genética , Lupus Eritematoso Sistémico/genética , Polimorfismo de Nucleótido Simple , Alelos , Estudios de Cohortes , Femenino , Genotipo , Haplotipos , Humanos , MasculinoRESUMEN
We obtained eight collections of DNA samples from ethnically matched systemic lupus erythematosus (SLE) patients and controls from five European countries totaling 783 patients and 1210 controls. A highly significant cline in the frequency of the PD1.3 A allele was found among controls but not among SLE patients. The frequency of the PD1.3 A allele increased from the Northeast to the Southwest of Europe. The cline was clearly apparent (P=1.2 x 10(-6)) when data from controls of other five SLE susceptibility studies were included in the analysis. This variation has severely biased SLE association studies owing to the lack of parallel changes in SLE patients. As a consequence, the PD1.3 A allele was more common in SLE patients than in controls in the Northeast and Center of Europe, similar to controls in Southeast Europe, and less frequent than in the controls in the Southwest of the Continent. This dissociation in allele frequencies between SLE patients and controls in different subpopulations indicated that programmed cell death 1 variation and disease susceptibility are not independent but the type of relationship is currently unclear. As allele frequency clines are common in other polymorphisms their impact in genetic epidemiology studies should be carefully considered.
Asunto(s)
Antígenos CD/genética , Proteínas Reguladoras de la Apoptosis/genética , Predisposición Genética a la Enfermedad/genética , Lupus Eritematoso Sistémico/epidemiología , Lupus Eritematoso Sistémico/genética , Polimorfismo Genético , Teorema de Bayes , Sesgo , Análisis por Conglomerados , Cartilla de ADN , Demografía , Europa (Continente)/epidemiología , Frecuencia de los Genes , Genotipo , Haplotipos/genética , Humanos , Receptor de Muerte Celular Programada 1RESUMEN
OBJECTIVE: To evaluate a contribution of selected laboratory parameters for a prediction of progressive and erosive development in patients with early rheumatoid arthritis (RA). METHODS: In a prospective study baseline levels of antibodies to cyclic citrullinated peptide (anti-CCP), IgM, IgA, and IgG rheumatoid factors (RFs) were measured by enzyme linked immunosorbent assay (ELISA) in 104 patients with RA with disease duration <2 years. Antikeratin antibodies (AKA) and antiperinuclear factor (APF) were detected by indirect immunofluorescence. Patients were divided into two groups based either on the presence or absence of erosions or according to progression of Larsen score at the end of the 24 months' follow up. RESULTS: Sixty seven (64%) patients developed radiographic erosions, 49 (47%) had progression in Larsen score, and 36 (35%) progressed by more than 10 Larsen units. Significant differences in erosions and progression between the two groups were detected for anti-CCP, AKA, APF, IgM RF, IgA RF, and IgG RF. Baseline Larsen score correlated significantly with anti-CCP, IgM RF, and IgA RF levels, and all measured antibodies correlated with the progression >10 units. The combination of anti-CCP and IgM RF increased the ability to predict erosive and progressive disease. CONCLUSION: The data confirmed that measurement of anti-CCP, AKA, APF, and individual isotypes of RFs was useful for prediction of structural damage early in the disease course. Combined analysis of anti-CCP and IgM RF provides the most accurate prediction.
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Artritis Reumatoide/inmunología , Autoanticuerpos/sangre , Anticuerpos Antinucleares/sangre , Artritis Reumatoide/diagnóstico por imagen , Biomarcadores/sangre , Progresión de la Enfermedad , Ensayo de Inmunoadsorción Enzimática/métodos , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Queratinas/inmunología , Péptidos Cíclicos/inmunología , Valor Predictivo de las Pruebas , Estudios Prospectivos , Radiografía , Factor Reumatoide/sangre , Índice de Severidad de la EnfermedadAsunto(s)
Colágeno Tipo I/genética , Predisposición Genética a la Enfermedad , Hiperostosis Esquelética Difusa Idiopática/genética , Polimorfismo de Longitud del Fragmento de Restricción , Receptores de Calcitriol/genética , Anciano , Cadena alfa 1 del Colágeno Tipo I , República Checa/epidemiología , ADN/análisis , Dermatoglifia del ADN , Femenino , Frecuencia de los Genes , Haplotipos , Humanos , Hiperostosis Esquelética Difusa Idiopática/epidemiología , Hiperostosis Esquelética Difusa Idiopática/patología , Masculino , Países Bajos/epidemiologíaRESUMEN
OBJECTIVE: To investigate the contribution of polymorphism in the immunoglobulin heavy chain variable region V1-69 gene set to genetic susceptibility to rheumatoid arthritis (RA) in Czech and British patients. METHODS: We used V1-69 gene sequence-specific polymerase chain reaction (PCR) and restriction enzyme digestion to study polymorphism in the V1-69 gene set in germline DNA of 109 Czech and 159 British RA patients and 164 ethnically matched controls. Polymorphism was further studied by nucleotide sequencing of the V1-69 gene locus in germline DNA. RESULTS: We found that all patients and controls had at least one V1-69 gene copy. In the Czech RA cohort, the dimorphic nucleotide in codon 73 of V1-69 (GAA or AAA) was present in the homozygous form 73(A/A) in 31 of 109 (28.4%) RA patients vs 12 of 79 (15.2%) controls [odds ratio (OR)=2.22, P<0.001]. When the RA patients and controls were classified according to HLA shared epitope (SE) status, 73(A/A) was found in 18 of 76 (23.7%) SE(+) patients compared with 13 of 38 (34.2%) SE(-) patients, four of 12 (18.2) SE(+) controls and eight of 57 (14%) SE(-) controls. This suggests that homozygosity for the dimorphic sequence 73(A) contributed to susceptibility to RA in SE(-) Czech individuals (OR=3.2, P<0.001). The most striking observation was that none of the 38 SE(-) Czech patients, compared with 11 of 76 (14.5%) SE(+) RA patients, three of 22 (13.6%) SE(+) and 11 of 57 (19.3%) SE(-) ethnically matched controls, were homozygous for the alternative dimorphic sequence 73(G/G) (OR=9.1, P<0.05). These data, however, were not replicated in a Caucasoid British RA population. CONCLUSION: The dimorphic sequence at codon 73 (73(A/A)) of the V1-69 gene contributes to genetic susceptibility in SE(-) Czech RA patients.
Asunto(s)
Artritis Reumatoide/genética , Genes de Inmunoglobulinas , Antígenos HLA-DR/genética , Cadenas Pesadas de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Polimorfismo Genético , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Artritis Reumatoide/inmunología , Epítopos/genética , Femenino , Predisposición Genética a la Enfermedad , Genotipo , Antígenos HLA-DR/inmunología , Cadenas HLA-DRB1 , Humanos , Masculino , Persona de Mediana EdadRESUMEN
OBJECTIVE: An increased incidence of allele 2 of the interleukin-1 receptor antagonist gene (IL1RN*2) in several inflammatory diseases has recently been reported. The aim of this study was to examine a variable number tandem repeat (VNTR) polymorphism of the IL1RN gene in patients with juvenile idiopathic arthritis (JIA). METHODS: Findings in 185 Czech patients with JIA were compared with those in 168 Czech controls, 50 JIA patients and 52 controls of Turkish origin, and 79 controls from central England. VNTR polymorphism analysis of IL1RN was performed by polymerase chain reaction using 2 flanking primers to amplify an 86-bp tandem repeat region in intron 2. RESULTS: The frequency and carriage rate of IL1RN*2 were significantly increased in Czech JIA patients compared with the Czech controls (frequency 27.6% versus 15.8%; carriage rate 44.3% versus 26.2%). Increased frequency and carriage rate of IL1RN*2 were found in 23.3% and 40.0% of Turkish JIA patients and in 17.3% and 34.6% of ethnically matched controls. The high representation of IL1RN*2 in 52.3% of the 22 patients with extended oligoarthritis, 31.3% of the 56 patients with enthesitis-related arthritis, and 42.9% of the 14 patients with other arthritis was particularly responsible for the increased frequency of IL1RN*2 in the Czech JIA patients. We found no association of IL1RN*2 with disease activity or severity parameters. CONCLUSION: Inheritance of IL1RN*2 may contribute to genetic susceptibility in several forms of autoimmune diseases, including JIA. The IL1RN*2 allele may be useful as a prognostic indicator of the evolution of an extended oligoarticular course of JIA.
Asunto(s)
Artritis Juvenil/genética , Sialoglicoproteínas/genética , Alelos , República Checa , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Proteína Antagonista del Receptor de Interleucina 1 , Secuencias Repetidas en TándemRESUMEN
We compared clinical data from 45 patients with Huntington's Disease (HD) with CAG triplet repeats and the planimetric measurement of the caudate nucleus head area (CNHA) in CT scans. The mean age of patients was 50.4 yrs (SD +/- 10.2), the mean duration of HD 7.4 yrs (4.6), the mean age at the onset of HD 43.1 yrs (11.1). HD started with motor symptoms in 28 patients, with psychiatric symptoms in 14 patients, the history was unknown in 3 patients. The paternal transmission was observed in 29 patients, the maternal one in 12 patients, unknown in 4 patients. The mean number of CAG repeats was 46.6 (6.1). The mean CNHA was 0.4 cm2 (0.1). We found statistically significant reversed correlation between CAG repeats and the age at the onset of HD (p < 0.0001, r -0.6). The earlier onset of HD in patients with the paternal transmission compared to the maternal one was found statistically significant (p < 0.05). This phenomenon was not related to the larger number of CAG triplets in patients with the paternal transmission. No differences either of the age at the onset of HD or numbers of CAG repeats were found between subgroups of HD patients starting with motor or psychiatric symptoms. We also observed the significant reversed correlation between the duration of HD and CNHA measurement (p < 0.001, r -0.5). Even in the earliest stage of HD patients showed the marked atrophy of CNHA.
Asunto(s)
Núcleo Caudado/diagnóstico por imagen , Enfermedad de Huntington/diagnóstico , Repeticiones de Trinucleótidos , Atrofia , Núcleo Caudado/patología , Femenino , Humanos , Enfermedad de Huntington/diagnóstico por imagen , Enfermedad de Huntington/genética , Enfermedad de Huntington/patología , Masculino , Persona de Mediana Edad , Tomografía Computarizada por Rayos XRESUMEN
Mutations, haplotypes, and other polymorphic markers in the phenylalanine hydroxylase (PAH) gene were analysed in 133 unrelated Czech families with classical phenylketonuria (PKU). Almost 95% of all mutant alleles were identified, using a combination of PCR and restriction analysis, denaturing gradient gel electrophoresis (DGGE), and sequencing. A total of 30 different mutations, 16 various RFLP/VNTR haplotypes, and four polymorphisms were detected on 266 independent mutant chromosomes. The most common molecular defect observed in the Czech population was R408W (54.9%). Each of the other 29 mutations was present in no more than 5% of alleles and 13 mutations were found in only one PKU allele each (0.4%). Four novel mutations G239A, R270fsdel5bp, A342P, and IVS11nt-8g-->a were identified. In 14 (5.1%) alleles, linked to four different RFLP/VNTR haplotypes, the sequence alterations still remain unknown. Our results confirm that PKU is a heterogeneous disorder at the molecular level. Since there is evidence for the gene flow coming from northern, western, and southern parts of Europe into our Slavic population, it is clear that human migration has been the most important factor in the spread of PKU alleles in Europe.
Asunto(s)
Alelos , Mutación , Fenilalanina Hidroxilasa/genética , Fenilcetonurias/genética , República Checa , Análisis Mutacional de ADN , Marcadores Genéticos , Genotipo , Haplotipos , Humanos , Fenilcetonurias/enzimología , Polimorfismo GenéticoRESUMEN
BACKGROUND: Phenylketonuria (PKU) is an autosomal recessive, disease, heterogeneous at the molecular level, caused by mutations in the gene of phenylalanine hydroxylase (PAH). The objective of the present work was to identify mutations and their frequency in the Central Bohemian and Prague population in relation to the clinical phenotype. METHODS AND RESULTS: The authors analyzed a group of 33 patients from 32 unrelated families. The phenotypic manifestations were classified as non-PKU hyperphenylalaninaemia (non-PKU-HPA), mild and classical PKU. Sixty-six mutant alleles of the PAH gene were analyzed by means of the polymerase chain reaction on a Perkin Elmer (480) apparatus and on PHC Techne. A total of eight mutations linked with five haplotypes were detected. R408W mutation linked with 2.4 haplotype was detected on 53% of mutant alleles. No type of mutation was detected by hitherto published procedures in 27% of mutant alleles. CONCLUSIONS: The finding on the distribution and frequency of mutations indicate a genotypic homogeneity of the PKU population in the Central Bohemian area and Prague and are consistent with hitherto published data from the Czech Republic. The revealed data can be used in prenatal and postnatal DNA diagnosis and genotype classification of PKU.
Asunto(s)
Mutación , Fenilalanina Hidroxilasa/genética , Fenilcetonurias/genética , Alelos , República Checa/epidemiología , Frecuencia de los Genes , Humanos , Fenilcetonurias/epidemiología , Reacción en Cadena de la PolimerasaRESUMEN
Twelve patients with bifocal intraocular lenses (3M, Suncoast and IOLAB) were evaluated. In 4 patients authors were not able to evaluate bifocality because of macular disorders and optic atrophy. In all remaining 8 patients the bifocality was strongly expressed. The main problem is a postoperative ammetropia. Implantation of bifocal lenses is connected with side effects which are not usually seen in implantations of monofocal intraocular lenses.