RESUMO
OBJECTIVE: To investigate serum levels of B cell activating factor (BAFF) in patients with myositis and correlate these to autoantibody profile, clinical phenotype and treatment. METHODS: BAFF levels in sera from 49 patients with dermatomyositis, 44 with polymyositis, 6 with inclusion body myositis and 30 matched controls were measured by ELISA. Specific autoantibodies were detected by line blot and western blot assays. RESULTS: Serum levels of BAFF were significantly higher in patients compared to healthy controls (p = 0.003). Patients with anti-Jo-1 autoantibodies had higher BAFF levels than control individuals (p<0.003) or patients without any specific autoantibodies (p<0.05). Patients with dermatomyositis had higher BAFF levels compared to polymyositis (p<0.05). Patients with interstitial lung disease (ILD) had higher BAFF levels than patients without ILD (p<0.05) or controls (p<0.01) but this could be explained by presence of anti-Jo-1 autoantibodies. BAFF levels correlated with serum creatine kinase (CK) (rs = 0.365, p = 0.0005) but not with C-reactive protein (CRP) levels. A negative correlation of BAFF levels with glucocorticoid daily dose for all patients (rs = -0.292, p = 0.003) and with cumulative glucocorticoid doses in early myositis cases (rs = -0.659, p<0.001) was recorded. CONCLUSION: Our finding of elevated serum levels of BAFF in patients with myositis with described phenotypes together with the correlations between levels of BAFF and CK and a negative correlation with dose of glucocorticoids, indicate that BAFF could be a potential therapeutic target in such cases.
Assuntos
Autoanticorpos/sangue , Fator Ativador de Células B/sangue , Miosite/sangue , Adolescente , Adulto , Idoso , Análise de Variância , Anticorpos Antinucleares/sangue , Autoanticorpos/imunologia , Proteína C-Reativa/análise , Estudos de Casos e Controles , Criança , Creatina Quinase/sangue , Dermatomiosite/sangue , Dermatomiosite/tratamento farmacológico , Dermatomiosite/imunologia , Feminino , Glucocorticoides/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Miosite/tratamento farmacológico , Miosite/imunologia , Polimiosite/sangue , Polimiosite/tratamento farmacológico , Polimiosite/imunologia , Estatísticas não Paramétricas , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/sangue , Adulto JovemRESUMO
BACKGROUND: Iron accumulation as seen in genetic haemochromatosis is a major cause of hepatic fibrogenesis. A link between chronic liver disease and Dupuytren's disease (DD) is well established, especially in alcoholics. AIM: The aim of the present study was to test the hypothesis that iron accumulation might cause fibrosis of the palmar aponeurosis leading to DD. PATIENTS AND METHODS: We examined iron metabolism, mutations of the HFE gene, serum cholesterol, alcohol consumption, presence of chronic liver disease, diabetes and history of severe manual work in a group of 90 patients who had undergone surgery for a severe form of DD. The tissue removed during surgery was histologically examined to confirm the diagnosis of DD. For a control group, we used 33 healthy subjects with similar profiles. RESULTS: The DD group consisted of 82 men and 8 women. Chronic liver disease was found in 27% of DD patients, compared with 6.1% of control subjects (P = 0.013). A history of hand traumatization was present in 33% of DD patients vs. 15% of control subjects (P = 0.048). Excessive alcohol consumption was present in 35.5% of DD patients compared with 15.1% of controls (P = 0.029). None of the other tested parameters, including the prevalence of HFE gene mutations, showed a significant difference between the two groups. CONCLUSIONS: Iron accumulation does not play a major role in the pathogenesis of DD. However, sex, age, manual labour and alcohol consumption are risk factors for progression of DD. We observed a high incidence of chronic liver disease in patients with DD.
Assuntos
Contratura de Dupuytren/etiologia , Contratura de Dupuytren/metabolismo , Ferro/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Consumo de Bebidas Alcoólicas/efeitos adversos , Colesterol/sangue , Diabetes Mellitus/metabolismo , Contratura de Dupuytren/genética , Contratura de Dupuytren/cirurgia , Feminino , Hemocromatose/complicações , Hemocromatose/genética , Hemocromatose/metabolismo , Humanos , Hepatopatias/complicações , Masculino , Pessoa de Meia-Idade , Mutação , Ocupações , Fatores de RiscoRESUMO
HFE-linked hereditary haemochromatosis is a common autosomal recessive disease among Caucasians. The primary pathogenetic mechanism is excessive absorption of iron, which is deposited in various organs with their subsequent damage. In 1996 the gene responsible for haemochromatosis was detected--the HFE gene and its major mutation C282Y. The discovery of further mutations followed. Two sites of point mutations in the HFE gene, C282Y and H63D, are associated with more than 80% of haemochromatosis cases. Another mutation-- S65C--was detected on 8% of chromosomes of haemochromatosis patients, which were negative for mutations C282Y or H63D. The objective of this study was to identify the allele frequency of S65C and other HFE mutations in the Czech population. DNA extracted from 481 randomly selected newborn screening cards (Guthrie cards) from all over the country was analysed by PCR-RFLP. No (0%) sample was identified as homozygous for S65C or C282Y mutation and 8 (1.67%) were homozygous for H63D mutation. Twelve (2.49%) samples were S65C heterozygous, 33 (6.86%) samples were C282Y heterozygous, and 128 (26.61%) were H63D heterozygous. Of these, 11 (2.29%) carried one copy of each mutation, i.e. were compound heterozygous. Two samples were S65C/H63D compound heterozygous and nine were C282Y/H63D compound heterozygous. Allele frequencies for S65C, C282Y, and H63D were 1.25% (95% CI, +/- 0.70), 3.43% (95% CI, +/- 1.15), and 14.97% (95% CI, +/- 2.25), respectively. The observed genotype frequency for S65C, C282Y, and H63D mutations in the Czech Republic agrees with those reported for other Central European populations.
Assuntos
Frequência do Gene , Hemocromatose/genética , Antígenos de Histocompatibilidade Classe I/genética , Proteínas de Membrana/genética , Mutação , República Tcheca/epidemiologia , Análise Mutacional de DNA , Predisposição Genética para Doença , Testes Genéticos , Hemocromatose/epidemiologia , Proteína da Hemocromatose , Humanos , Recém-Nascido , Triagem NeonatalRESUMO
BACKGROUND: Hereditary hemochromatosis is one of the most common autosomal recessive diseases. Aim of the study. 1. To establish frequency of C282Y and H63D mutations in the HFE gene (the hemochromatosis gene) in general population of the Czech Republic and in patients with hemochromatosis. 2. To find out whether hemochromatosis in homo- or heterozygous state plays a role in the pathogenesis of rheumatic diseases. METHODS AND RESULTS: In 32 patients with hereditary hemochromatosis, in 84 patients with polymyositis or dermatomyositis, in 246 patients with juvenile idiopathic arthritis and in 481 persons of the control group the presence of HFE gene mutations was etablished. The HFE gene mutations were screened for by restriction enzyme analysis performed on PCR amplified products. In the control group, 6.86% carriers of the C282Y mutation and 26.61% those of H63D were found. Homozygous C282Y or H63D mutation was found in 90.6% (p<0.001) of patients with hemochromatosis. Heterozygous C282Y mutation was found in 12.2% (p<0.05) of patients with juvenile idiopathic arthritis. We didn't detected higher prevalence of HFE gene mutations in patients with polymyositis and dermatomyositis. CONCLUSIONS: Results of this study show that heterozygosity for C282Y mutation may be a risk factor for juvenile idiopathic arthritis but not for polymyositis and dermatomyositis.
Assuntos
Predisposição Genética para Doença , Antígenos de Histocompatibilidade Classe I/genética , Proteínas de Membrana/genética , Mutação , Doenças Reumáticas/genética , Frequência do Gene , Hemocromatose/genética , Proteína da Hemocromatose , Heterozigoto , Homozigoto , Humanos , Fatores de RiscoRESUMO
The aim of this study was to assess the rate of hepatitis B virus (HBV) and hepatitis C virus (HCV) coinfection ("the coinfection") in chronic liver disease (CLD) and to reveal overt and hidden HBV infection in patients with antibodies to HCV (anti-HCV). A total of 209 untreated patients (64 with chronic hepatitis B, 79 with chronic hepatitis C and 66 with porphyria cutanea tarda (PCT)) were screened for serological markers of HBV and HCV infection in serum by third generation enzyme-linked immunosorbent assay (ELISA) methods and for HBV DNA and HCV RNA in serum by polymerase chain reaction (PCR). The rate of the overt coinfection in chronic hepatitis B was very low (2/64, 3%). However, in chronic hepatitis C, the rate of the hidden coinfection with HBV was relatively high (19/79, 24%); these patients had higher alanine transaminase (ALT) and asparagine transaminase (AST) levels in serum and a more advanced liver disease. In PCT patients, the rates of HBV and HCV infections were the same, 21% (14/66). In the PCT patients infected with HBV or HCV, the rate of the coinfection was 33% (7/21). The PCT patients with the coinfection had a high serum ALT level and the worst histological picture in the liver. The hidden HBV infection was more frequent than the overt one. The possibility of the overt or hidden coinfection in CLD renders a detailed analysis of all serum samples for both viruses mandatory. Vaccination against HBV infection should be offered to anti-HCV-positive individuals as well as to PCT patients not showing antibodies to HBV (anti-HBV).
Assuntos
Hepatite B Crônica/epidemiologia , Hepatite C Crônica/epidemiologia , Porfiria Cutânea Tardia/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença Crônica , Comorbidade , República Tcheca/epidemiologia , DNA Viral/análise , Ensaio de Imunoadsorção Enzimática , Feminino , Hepacivirus/isolamento & purificação , Anticorpos Anti-Hepatite B/sangue , Vírus da Hepatite B/isolamento & purificação , Hepatite B Crônica/sangue , Anticorpos Anti-Hepatite C/sangue , Hepatite C Crônica/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Porfiria Cutânea Tardia/sangue , RNA Viral/análiseRESUMO
A commercially available set, the Gen-Probe amplified MTD test (Mycobacterium tuberculosis Direct Test; Gen-Probe Incorporated, 9080 Campus Point Drive, 92121 California, USA) has been applied for the detection of the M. tuberculosis complex in clinical material in parallel to direct microscopy and cultivation in liquid and solid growth media. The method is based on the amplification of a specific 16 S rRNA segment. Detection of the amplified segment is facilitated by single-strand probes tagged with acridinium ester. Hybridization results are interpreted in RLU (Relative Light Unit) values and readings exceeding 30,000 RLU are considered positive, lower values being negative. In all, there have examined 69 samples of clinical material (35 sputums, 29 bronchoalveolar washings or bronchial aspirations, and 5 cerebrospinal fluids). As positive amplification controls served freshly cultured strains of M. tuberculosis and M. bovis BCG; negative controls were M. gordonae, M. xenopi, M. kansasii, and M. terrae. Out of a total 69 samples examined, M. tuberculosis was detected in 28 (40.7%) cases by the Gen-Probe amplified MTD test, in 10 (14.5%) cases by direct microscopy, and in 26 (37.7%) cases by cultivation. In samples from 8 patients M. tuberculosis was detected by just Gen-Probe amplified MTD test the results of cultivation and direct microscopy being negative. Culture-positive samples yielded no Gen-Probe amplified MTD test negative results. The Gen-Probe amplified MTD test contributes significantly to the speeding up of diagnostics in tuberculosis because the results is obtainable with six hours. Under the conditions of this pilot study the results were termed as preliminary and collaborating physicians were advised to wait for cultivation results and their definite evaluation.
Assuntos
Mycobacterium tuberculosis/genética , Sondas RNA , Líquido da Lavagem Broncoalveolar/microbiologia , Amplificação de Genes , Humanos , Mycobacterium tuberculosis/citologia , Mycobacterium tuberculosis/crescimento & desenvolvimento , Hibridização de Ácido Nucleico , Escarro/microbiologiaRESUMO
The epidemiological relationship between tuberculosis cases in a prison and between cases within five families was investigated. Therefore, the isolated Mycobacterium tuberculosis strains were subjected to restriction fragment length polymorphism (RFLP) analysis using insertion sequence IS6110 as a probe. In case of 11 patients, the expected links of transmission were confirmed by RFLP typing. In contrast, in case of 4 patients the conclusion of classical contact tracing were not in agreement with the DNA fingerprinting results. These findings reinforce the usefulness of this recently developed technique as an additional tool in contact tracing. The IS6110DNA fingerprints of all strains investigated consisted of 7 to 13 bands and showed a high degree of polymorphism. Comparison of these fingerprints with those recorded in the Czech Republic previously, revealed the presence of a predominant DNA fingerprint type, without a known connection between the cases. Furthermore, other patterns found in the present study showed a high degree of similarity with the previously obtained fingerprints. Part of the patients were sampled twice. All of these double isolates showed identical fingerprints, confirming the previously described stability of IS6110DNA fingerprints. In contrast, from one couple of strains, isolated from husband and wife both suffering from tuberculosis, a slight change in one of the two patterns was observed. The patterns shared 10 band positions, confirming the expected relationship between these cases, but one of the patterns contained one additional band.
Assuntos
Busca de Comunicante/métodos , Impressões Digitais de DNA , Mycobacterium tuberculosis/genética , Tuberculose/transmissão , República Tcheca , Feminino , Humanos , Masculino , Polimorfismo de Fragmento de RestriçãoRESUMO
BACKGROUND: Anaemia is a common phenomenon encountered in patients on hemodialysis. Although treatment with rHuEPO therapy is effective, it may fail even at high doses. As rHuEPO efficacy depends on the bioavailability of iron, we monitored the effect of consistent iron supplementation on hematocrit levels and rHuEPO dosage. METHODS AND RESULTS: 24 patients of our outpatient dialysis centre were included in this study. The mean age was 59 years. The age group over 60 included 14 patients. The mean duration of dialysis treatment was 23.8 months. The patients were followed for 6 months according to the NKF-DOQI (National Kidney Foundation Dialysis Outcomes Quality Initiative) recommendations for the treatment of anaemia. Following values were examined monthly: hematocrit, transferin saturation (TSAT) and ferritin. Iron and rHuEPO dosage was adjusted accordingly. Genetic tests for haemochromatosis were conducted in 4 patients with the highest value of TSAT and ferritin. TSAT increased from a mean of 15.9% to 35.9% (p < 0.001). In 23 patients (96%) TSAT levels were within the recommended range after the treatment. Hematocrit increased from 27.7% to 35.7% (p < 0.001). The recommended value of 33% was achieved in 18 patients (75%). The weekly dose of eHuEPO fell from 3958 IU (International Unit) to 2042 IU (p < 0.001), i.e. 1857 IU of rHuEPO were saved per week, per patient. The average dose of iron administered was 157 mg per week. The average level of ferritin rose from 457 micrograms/k to 1387 micrograms/l (p < 0.001). All results were comparable, even in the group of the senior's selected cases. Genetic testing for haemochromatosis showed mutation H63D in heterozygous state of HFE gene in 2 of 4 patients with the highest value of TSAT and ferritin. Sufficient iron supplementation leads to a significant rise in hematocrit and a concomitant decrease of required rHuEPO doses. TSAT, and not ferritin, is a good marker of iron bioavailability. CONDITIONS: The financial savings due to decreased rHuEPO requirements are 20 times higher than the costs related to iron supplementation, calculated in relation to prices valid for the Czech Republic in 1999. Cause and effect of increased level of ferritin should be carefully studied.
Assuntos
Anemia/terapia , Eritropoetina/uso terapêutico , Ferro/uso terapêutico , Compostos Organometálicos/uso terapêutico , Polissacarídeos/uso terapêutico , Diálise Renal , Adulto , Idoso , Anemia/sangue , Anemia/economia , Anemia/etiologia , Redução de Custos , República Tcheca , Eritropoetina/economia , Feminino , Hematócrito , Humanos , Ferro/sangue , Ferro/economia , Masculino , Pessoa de Meia-Idade , Compostos Organometálicos/economia , Polissacarídeos/economia , Proteínas Recombinantes , Diálise Renal/efeitos adversos , Transferrina/análiseRESUMO
Using commercially available probes Gen-Probe (Accu-Probe, Epignost, Austria) labelled with acridinium ester, the authors identified 102 cultures of mycobacteria which are the causal agents of classical tuberculosis or cause mycobacterioses which are difficult to control by therapeutic measures. The cultures were isolated from sputum or lymph nodes and cultivated on Löwenstein-Jensen medium. The authors proved a 100% sensitivity of the probe for isolates of M. tuberculosis and M. kansasii. In a group of 36 strains sent as suspect M. avium 33 strains reacted with the probe for the M. avium complex, 28 strains with the M. avium probe and 4 strains with the M. intracellulare. One isolate hybridized only with the probe for the M. avium complex. In one isolate a positive hybridization value was recorded with the M. gordonae probe and in one pigmented strain with the M. kansasii probe. One isolate did not hybridize with any of the probes used. In one instance a mixed culture of M. tuberculosis and M. avium was involved. The results of hybridization were interpreted in RLU values (relative light unit). Isolates above 30,000 RLU were evaluated positively, as recommended by the manufacturer.