RESUMO
BACKGROUND: Recently, a new approach of autologous chondrocyte implantation technique (using as biomaterial a collagen type i/iii membrane) based on increasing cell density called HD-ACI (High Density Autologous Chondrocyte Implantation) has been described. The objective of this paper was to study the clinical outcome and incidence of subchondral bone oedema in patients with cartilage lesions in the knee treated with HD-ACI at 1-2 years of follow-up. METHODS: This is a retrospective study performed with forty patients with chondral injuries grade iii-iv. All patients were treated with HD-ACI with a cellular dose of 5×106 chondrocytes /cm2 of lesion. The subjective perception of improvement of symptoms and functionality was measured with the IKDC score (International Knee Documentation Committee). The presence of bone oedema was assessed at 6, 12 and 24 months of follow-up by magnetic resonance imaging. RESULTS: IKDC values showed a significant improvement at 12 and 24 months (P<.001). The mean difference of IKDC between the baseline visit and 12 months was 26.3 points, and 31.6 points at 24 months. Twenty-seven point five percent of the patients presented subchondral bone oedema at 2 years of follow-up. CONCLUSIONS: HD-ACI is an effective and safe treatment that improves pain, clinical perception and functionality of the joint. No correlation was found between the presence of bone oedema and the patients' clinical outcome.
Assuntos
Doenças Ósseas/etiologia , Condrócitos/transplante , Edema/etiologia , Articulação do Joelho/cirurgia , Complicações Pós-Operatórias/etiologia , Adulto , Feminino , Seguimentos , Humanos , Masculino , Estudos Retrospectivos , Fatores de Tempo , Transplante Autólogo/efeitos adversos , Transplante Autólogo/métodosRESUMO
OBJECTIVE: The aim of this study is to present our initial experience with the stereotactic echo-MRI fusion system for diagnosing prostate cancer. MATERIAL AND METHODS: Between September 2014 and January 2015, we performed 50 prostate biopsies using the stereotactic echo-MRI fusion system. The 3-Tesla multiparameter MR images were superimposed using this image fusion system on 3D echo images obtained with the Biopsee system for the exact locating of areas suspected of prostate cancer. The lesions were classified using the Prostate Imaging Report and Date System. RESULTS: We assessed a total of 50 patients, with a mean age of 63 years (range, 45-79), a mean prostate-specific antigen level of 8 ng/mL (range, 1.9-20) and a mean prostate volume of 52mL (range, 12-118). Prostate cancer was diagnosed in 69% of the patients and intraepithelial neoplasia in 6%. The results of the biopsy were negative for 24% of the patients. The results of the biopsy and MRI were in agreement for 62% of the patients; however, 46% also had a tumour outside of the suspicious lesion. We diagnosed 46% anterior tumours and 33% apical tumours. One patient had a haematuria, another had a haematoma and a third had acute urine retention. CONCLUSIONS: Multiparametric prostatic MRI helps identify prostate lesions suggestive of cancer. The Biopsee echo-MRI fusion system provides for guided biopsy and increases the diagnostic performance, reducing the false negatives of classical biopsies and increasing the diagnosis of anterior tumours. Transperineal access minimises the risk of prostatic infection and sepsis.
Assuntos
Imageamento por Ressonância Magnética , Próstata/patologia , Neoplasias da Próstata/patologia , Ultrassonografia , Idoso , Humanos , Biópsia Guiada por Imagem/métodos , Masculino , Pessoa de Meia-Idade , Imagem Multimodal , PeríneoRESUMO
BACKGROUND: Hepatitis C virus replicates by the synthesis of an antigenomic HCV-RNA. As the end point of anti-viral therapy is to decrease viral replication, the amount of antigenomic HCV-RNA could influence the response. AIM: To study if amounts of genomic and antigenomic HCV-RNA in the baseline liver biopsy are predictive factors of response to anti-viral therapy. METHODS: Eighty-eight patients with chronic HCV infection (anti-HIV-negative) treated with pegyltaed-interferon-alpha2b plus ribavirin for 12 months were included. Intrahepatic genomic and antigenomic HCV-RNA concentrations were determined by real-time polymerase chain reaction and percentage of infected hepatocytes by in situ hybridization. RESULTS: Of the 88 patients, 31% were responders while 69% were not. Median of antigenomic HCV-RNA in liver of responders and non-responders was 120 000 copies/microg RNA (range: 10,000-775,000) vs. 150,000 copies/microg RNA (range: 100-3,200,000; P = 0.38). Median of genomic HCV-RNA in liver of responders was 1,250,000 copies/microg RNA (range: 5000-9,000,000) and in non-responders 3,180,000 copies/microg RNA (range: 4600-18,000,000; P = 0.0191). Predictive factors of response in the logistic regression were: intrahepatic amount of genomic HCV-RNA, percentage of infected hepatocytes and previous therapy. CONCLUSION: Response to 12 months of therapy with pegylated interferon-alpha2b plus ribavirin depends on the amount of genomic HCV-RNA in the pre-treatment liver biopsy.
Assuntos
Antivirais/uso terapêutico , Genoma Viral/efeitos dos fármacos , Hepatite C Crônica/tratamento farmacológico , Interferon-alfa/uso terapêutico , Ribavirina/uso terapêutico , Adulto , Biópsia/métodos , Quimioterapia Combinada , Feminino , Genoma Viral/genética , Hepatite C Crônica/genética , Humanos , Interferon alfa-2 , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Polietilenoglicóis , RNA Viral/genética , Proteínas RecombinantesRESUMO
We have recently described the presence of occult hepatitis C virus (HCV) infection (HCV-RNA in liver in the absence of anti-HCV and serum HCV-RNA) in patients with persistently abnormal liver function tests of unknown aetiology. The aim of this study was to compare the characteristics of patients with occult HCV infection vs those of patients with chronic hepatitis C. We compared clinical features of 68 patients with occult HCV infection and 69 untreated chronic HCV patients (anti-HCV and serum HCV-RNA positive), matched for age, gender, duration of abnormal liver function tests and body mass index. Aspartate aminotransferase and alanine aminotransferase were higher (P < 0.001) in chronic HCV, but cholesterol and triglycerides were significantly higher in patients with occult HCV infection (P < 0.001 and P = 0.002). Chronic HCV patients had higher gamma-globulin (P = 0.005), alpha-foetoprotein (P < 0.001) and iron (P < 0.001) levels. Percentage of patients with necroinflammatory activity and fibrosis was higher (P < 0.001) in chronic HCV than in occult HCV infection. Mean percentage of infected hepatocytes was higher (P = 0.001) in chronic HCV (10.1%) than in occult HCV infection (5.3%). This occult HCV infection is a milder disease than chronic HCV, and this could be related to the significantly lower number of infected hepatocytes observed in occult HCV.
Assuntos
Hepacivirus/crescimento & desenvolvimento , Hepatite C Crônica/sangue , Hepatite C/sangue , Adulto , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Biópsia , Colesterol/sangue , Feminino , Ferritinas/sangue , Hepatite C/fisiopatologia , Hepatite C/virologia , Hepatite C Crônica/fisiopatologia , Hepatite C Crônica/virologia , Humanos , Ferro/sangue , Masculino , Pessoa de Meia-Idade , RNA Viral/sangue , Triglicerídeos/sangue , alfa-Fetoproteínas/metabolismo , gama-GlutamiltransferaseRESUMO
BACKGROUND: Occult hepatitis C virus infection is defined by the presence of hepatitis C virus-RNA in liver but with undetectable anti-hepatitis C virus and serum viral RNA. AIM: To study the response to anti-viral therapy in occult hepatitis C virus infection to assess the pathogenic effect of occult hepatitis C virus. METHODS: Ten patients with occult hepatitis C virus infection were treated with pegylated-interferon plus ribavirin for 24 weeks and were followed-up 24 weeks after therapy. All patients had abnormal alanine aminotransferase, hepatitis C virus-RNA positive in peripheral blood mononuclear cells and liver necroinflammation. RESULTS: At the end of treatment and follow-up, the percentage of patients with normal alanine aminotransferase was 80% (95% CI: 48-96%) and 60% (95% CI: 31-84%) respectively, and hepatitis C virus-RNA in peripheral blood mononuclear cells was negative in 80% (95% CI: 48-96%) and 70% (95% CI: 40-90%) cases. At the end of follow-up sustained response was observed in 30% (95% CI: 11-61%) of cases. Five patients underwent a second liver biopsy. In all cases, liver hepatitis C virus-RNA persisted, although hepatitis C virus-RNA load was significantly lower (3.2 x 10(4) +/- 5.1 x 10(4) copies/microg RNA) than in the basal biopsy (2.4 x 10(5) +/- 3.8 x 10(5) copies/microg RNA); (P = 0.043). Necroinflammation and fibrosis decreased in three cases. CONCLUSION: The biochemical, virological and histological response to therapy achieved in patients with occult hepatitis C virus infection demonstrates the pathologic effects of occult hepatitis C virus.
Assuntos
Antivirais/uso terapêutico , Hepacivirus , Hepatite C/tratamento farmacológico , Interferon-alfa/uso terapêutico , Fígado/virologia , Polietilenoglicóis/uso terapêutico , Ribavirina/uso terapêutico , Adulto , Alanina Transaminase/sangue , Intervalos de Confiança , Feminino , Seguimentos , Hepacivirus/genética , Hepacivirus/patogenicidade , Hepatite C/enzimologia , Hepatite C/virologia , Humanos , Hibridização In Situ/métodos , Interferon alfa-2 , Leucócitos Mononucleares/virologia , Fígado/patologia , Cirrose Hepática/enzimologia , Cirrose Hepática/virologia , Masculino , Pessoa de Meia-Idade , RNA Viral/análise , Proteínas Recombinantes , Estatísticas não Paramétricas , Carga ViralRESUMO
In this work, we have shown that hepatitis C virus (HCV) and hepatitis B virus (HBV) can coexist in the same hepatocyte using double fluorescent in situ hybridization in liver biopsy samples from patients with chronic HCV infection with occult HBV infection. Digital image analysis of hybridization signals showed that the HBV DNA levels in coinfected hepatocytes were lower than those in cells infected only with HBV. This finding supports the hypothesis of inhibition of HBV replication by HCV. Furthermore, HCV RNA levels were lower in coinfected cells than in cells infected only with HCV, suggesting that HBV may also inhibit HCV replication.
Assuntos
Hepacivirus/isolamento & purificação , Vírus da Hepatite B/isolamento & purificação , Hepatite B/complicações , Hepatite C Crônica/complicações , Hepatócitos/virologia , Biópsia , DNA Viral/análise , Hepacivirus/genética , Hepatite B/virologia , Vírus da Hepatite B/genética , Humanos , Fígado/citologiaRESUMO
BACKGROUND: Occult hepatitis C virus (HCV) infection is characterised by the presence of HCV-RNA in the liver in the absence of anti-HCV, and serum viral RNA. Up to 70% of these patients also have HCV-RNA in peripheral blood mononuclear cells (PBMC) but it is not known if HCV is replicating in these cells. AIM: We studied possible HCV replication in PBMC of 18 patients with an occult HCV infection who were selected on the basis of HCV-RNA positivity in PBMC. METHODS: Detection of HCV-RNA positive and negative strands in PBMC was done by strand specific reverse transcriptase-polymerase chain reaction (RT-PCR) and by in situ hybridisation. RESULTS: The presence of HCV-RNA positive strand in PBMC was confirmed in all patients by strand specific RT-PCR and by in situ hybridisation. Mean percentage of PBMC which had the HCV-RNA positive strand was 3.3% (95% confidence interval (CI) 2.1-4.4) The HCV-RNA negative strand was found in the PBMC of 11/18 (61%) patients by strand specific RT-PCR and confirmed by in situ hybridisation, and the percentage of PBMC harbouring the HCV-RNA negative strand was 3.1% (95% CI 0.8-5.5). There was a significant correlation (p = 0.001, r = 0.84) between the percentage of PBMC with the HCV-RNA positive strand and that of PBMC with the HCV-RNA negative strand. CONCLUSION: HCV replicates in the PBMC of patients with occult HCV infection and thus, although these patients do not have serum HCV-RNA, they could be potentially infectious.
Assuntos
Portador Sadio/virologia , Hepacivirus/fisiologia , Hepatite C Crônica/virologia , Leucócitos Mononucleares/virologia , Replicação Viral , Adulto , Feminino , Humanos , Hibridização in Situ Fluorescente , Masculino , Pessoa de Meia-Idade , RNA Viral/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e EspecificidadeRESUMO
Epidemiological studies have demonstrated a correlation between oral lichen planus and different liver diseases. The new virus termed TT virus (TTV) is highly prevalent in patients with chronic hepatitis of different etiology and it may be speculated that TT virus may be involved in the pathogenesis of oral lichen planus. This study examined the presence of TT virus DNA in serum by PCR and in oral mucosa biopsies by in situ hybridization from 20 patients with oral lichen planus (13 with chronic hepatitis and seven without liver disease). Serum and oral mucosa biopsies from six patients all with chronic hepatitis with leukoplakia were also studied as controls. TT virus DNA was positive in the serum of 17/20 (85%) of the patients with oral lichen planus and in all the controls. TT virus DNA hybridization signals were detected in mucosa biopsies from all the patients with TT virus DNA in serum but in none of the three cases without this marker. The percentage of positive cells ranged from 1.6-80%. No differences were found in the percentage of positive cells between TT virus positive patients with and without oral lichen planus and there was no relationship between the number of positive cells and the intensity of the inflammatory infiltrate. In conclusion, TT virus infects oral epithelial cells but the results do not support a role for TT virus in causing oral lichen planus.
Assuntos
Infecções por Vírus de DNA/virologia , Líquen Plano/virologia , Mucosa Bucal/virologia , Torque teno virus/isolamento & purificação , Idoso , Infecções por Vírus de DNA/sangue , DNA Viral/sangue , Feminino , Humanos , Hibridização In Situ , Líquen Plano/sangue , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Torque teno virus/genéticaRESUMO
Patients with chronic renal failure on hemodialysis have a high risk of infections with viruses such as hepatitis B (HBV), hepatitis C (HCV), GB virus C/hepatitis G (GBV-C/HGV) and TT (TTV) viruses. The prevalence of HBV, HCV, GBV-C/HGV and TTV in patients with chronic renal failure who are on conservative management before entering into a hemodialysis program (predialysis) in comparison with hemodialyzed patients was studied to elucidate whether the high prevalence of these viruses is influenced by that observed in the predialysis stage. The presence of hepatitis B virus surface antigen (HBsAg), HCV RNA, GBV-C/HGV RNA and TTV DNA was analyzed in sera from 80 patients with chronic renal failure (35 on predialysis and 45 on hemodialysis). HBsAg, HCV RNA, GBV-C/HGV RNA and TTV DNA were detected in one (2.8%), six (17.1%), eight (22.5%) and 16 (45.7%) of the 35 patients on predialysis. Two (5.7%) of these patients were coinfected with HCV and GBV-C/HGV, whereas six (17.1%) had GBV-C/HGV and TTV coinfection. In the 45 hemodialyzed patients, HBsAg, HCV RNA, GBV-C/HGV RNA and TTV DNA were detected in one (2.2%), two (4.4%), seven (15.5%) and 26 (57.7%). One (2.2%) patient had HBV and TTV coinfection, two (4.4%) HCV and TTV coinfection whereas four (8.8%) were coinfected with GBV-C/HGV and TTV. No differences regarding age, gender, previous surgery and number of transfusions were found between infected and uninfected patients within and between both groups. In conclusion, the prevalence of the viruses studied in predialysis may influence their prevalence in dialysis units.
Assuntos
Flaviviridae/isolamento & purificação , Hepacivirus/isolamento & purificação , Vírus da Hepatite B/isolamento & purificação , Hepatite Viral Humana/virologia , Torque teno virus/isolamento & purificação , Idoso , DNA Viral/análise , Feminino , Antígenos de Superfície da Hepatite B/análise , Hepatite Viral Humana/transmissão , Humanos , Falência Renal Crônica/complicações , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , RNA Viral/análise , Diálise RenalRESUMO
The main site of TT virus (TTV) replication remains unknown. Therefore, we have studied the presence and titres of TTV DNA in paired serum, liver and PBMC samples from 50 patients with liver disease (32 with chronic hepatitis B or C, seven with cryptogenic hepatitis and 11 with nonviral liver disease) were included. TTV DNA was analysed by polymerase chain reaction (PCR) using primers from the open reading frame 1 (ORF 1) and from the untranslated region (UTR) and titres were semiquantified by PCR using an external standard. TTV DNA was detected in 26% of serum, 24% of liver and 14% of PBMC samples with ORF 1 primers. When UTR primers were used, 70% of serum and liver samples and 64% of PBMC were TTV DNA positive. No differences between TTV positive and negative patients were found regarding epidemiological or biochemical parameters. Trypsin treatment and fluorescent in situ hybridization confirm the intracellular location of TTV in PBMC. The mean of TTV DNA titres was statistically higher in liver than in serum or PBMC. TTV titres in serum correlated with those in PBMC but not with those in liver. In conclusion, although the liver seems to be the main site for TTV replication, this virus is also able to infect PBMC.
Assuntos
Infecções por Vírus de DNA/virologia , DNA Viral/sangue , Hepatite Crônica/virologia , Fígado/virologia , Torque teno virus/isolamento & purificação , Adulto , DNA Viral/análise , Feminino , Hepatite Crônica/etiologia , Humanos , Hibridização in Situ Fluorescente , Leucócitos Mononucleares/virologia , Masculino , Pessoa de Meia-Idade , Filogenia , Reação em Cadeia da PolimeraseRESUMO
To study the existence of GB virus C/hepatitis G virus (GBV-C/HGV) variants with different tropism, we have analyzed the heterogeneity and quasispecies composition of GBV-C/HGV isolated from in vitro-infected peripheral blood mononuclear cells (PBMC) and from sera, livers, and PBMC from two chronically infected patients. For this purpose, the GBV-C/HGV 5' noncoding region (5'NCR) was amplified by reverse transcription-PCR and the amplified products were cloned and sequenced. These analyses showed that the master 5'NCR sequences isolated from the in vitro-infected PBMC and from the PBMC isolated from the patient whose serum was used as the inoculum were identical but different from that of the inoculum. Furthermore, phylogenetic analysis revealed that all PBMC sequences grouped together into a branch which was separate from those of the inoculum. For one of the two chronically infected patients, all the sequences from the PBMC and one from the liver clustered into a single branch while the sequences from the serum and all the other liver sequences grouped together in the other branch. For the other patient, the sequences from the serum and PBMC and three sequences from the liver grouped together into one branch, while the remaining five sequences from the liver were separated in a different cluster. In conclusion, our results support the existence of different GBV-C/HGV variants with different tissue tropism.
Assuntos
DNA Viral/genética , Flaviviridae/classificação , Hepatite Viral Humana/virologia , Adulto , Sequência de Bases , Doença Crônica , DNA Viral/análise , Flaviviridae/genética , Flaviviridae/imunologia , Humanos , Soros Imunes , Leucócitos Mononucleares/virologia , Fígado/virologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNARESUMO
A novel hepatitis-associated virus named TT virus (TTV) has been isolated. However, its hepatotropism has not been proven. We have retrospectively analyzed the presence of TTV-DNA by polymerase chain reaction (PCR) and in situ hybridization in liver biopsies from 30 patients with liver disease (15 TTV-DNA-positive and 15 TTV-DNA-negative in serum), and prospectively in serum and liver from eight patients with normal liver histology. TTV-DNA was detected by PCR in the liver from the 15 patients with serum TTV-DNA and in serum and liver of two of the eight patients without liver disease. TTV-DNA titers in liver were 10 times higher than in serum, although no correlation between TTV-DNA titers in serum and liver were observed. In situ hybridization shows positive signals in the hepatocytes of the 17 patients infected by TTV but in none of the TTV-DNA-negative patients by PCR. No morphological changes were observed in the hepatocytes showing hybridization signals. The percentage of positive hepatocytes ranged from 2.1% to 30% and correlated with the TTV-DNA titers in liver (r = 0.54; P = 0.037). In conclusion, our results show that TTV is able to infect liver cells although they do not support a role for TTV in causing liver disease.
Assuntos
Vírus de DNA/genética , DNA Viral/isolamento & purificação , Fígado/virologia , Adulto , Sequência de Bases/genética , Biópsia , Circoviridae/genética , Feminino , Humanos , Hibridização In Situ , Fígado/patologia , Hepatopatias/patologia , Hepatopatias/virologia , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: A novel virus named TT virus (TTV) has been isolated recently from patients with posttransfusional hepatitis of unknown etiology. The prevalence of TTV in several groups at risk has been reported, however, there is no information about the prevalence of TTV in patients on continuous ambulatory peritoneal dialysis (CAPD) without blood transfusions or hemodialysis antecedents. OBJECTIVE: To study the incidence of TTV in serum and peripheral blood mononuclear cells (PBMC) of CAPD patients. DESIGN: TTV DNA was detected by polymerase chain reaction, using primers from the open reading frames (ORF) 1 and 2, in serum and PBMC from 22 CAPD patients who had not received blood transfusions or hemodialysis therapy prior to CAPD. As controls, sera from 20 patients with chronic viral hepatitis (10 with HBV and 10 with HCV) and 20 healthy donors were included in the study. RESULTS: TTV DNA was detected in the serum of 5 of 22 (22.7%) CAPD patients with both sets of primers. Four of the 5 (80%) patients with TTV DNA in their serum were TTV positive in their PBMC with primers from ORF1 and ORF2. Five of 20 (25%) patients with chronic viral hepatitis (2 patients with HBV and 3 with HCV) and 4 of 20 (20%) healthy donors were TTV DNA positive in serum. No relation was found between TTV infection and the underlying kidney disease, previous surgery, and abnormal alanine aminotransferase levels. CONCLUSION: We have found a relatively high prevalence of TTV that is similar to that found in healthy donors and in patients with chronic viral hepatitis.
Assuntos
Sangue/virologia , Vírus de DNA/isolamento & purificação , Leucócitos Mononucleares/virologia , Diálise Peritoneal Ambulatorial Contínua , Adulto , Idoso , Vírus de DNA/genética , DNA de Cadeia Simples , Feminino , Hepatite C Crônica/virologia , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , PrevalênciaRESUMO
Twenty-four patients with chronic hepatitis B virus (HBV), antibody to hepatitis B e antigen (anti-HBe), HBV DNA positivity, and alanine transaminase (ALT) elevation who failed previous interferon alfa (IFN-alpha) therapy were included in a pilot study of combination therapy with ribavirin and IFN-alpha. The patients received daily oral ribavirin (1,000-1,200 mg according to body weight) plus 5 million units (MU) IFN-alpha2b three times a week for 12 months and were followed-up for 12 months. The median viremia level decreased significantly at the end of treatment (1.2 x 10(3) copies/mL) and follow-up (4.0 x 10(2) copies/mL) compared with the baseline (3.0 x 10(6) copies/mL; P <.05). After 12 months, 8 of 24 (33%) patients had cleared HBV DNA and 12 (50%) had normal ALT levels. At the end of the study virological and biochemical response was 50% and 21%, respectively. Thus, virological and biochemical response sustained in 5 of 24 (21%) patients retreated with ribavirin and IFN-alpha; none of them lost hepatitis B surface antigen (HBsAg). Liver histology improved in 2 of 4 sustained responders but in none of the 12 nonresponders with paired biopsies (P =.05). The response was independent of dose and duration of previous treatment, viral load, or the distribution of HBV precore wild-type/mutant variants. However, sustained responders had significantly higher necroinflammation (P =.036) and fibrosis (P =.007) scores. IFN-alpha-related side effects were mild and reversible on discontinuation. In 4 (17%) patients who suffered nausea and diarrhea the ribavirin dosage was reduced by 50% after 1 month of therapy and finally discontinued in all of them. No patient had liver disease decompensation. In summary, combination therapy with ribavirin and IFN-alpha may be efficacious to treat viremic anti-HBe-positive patients with chronic hepatitis B who have failed previous IFN therapy.
Assuntos
Antivirais/uso terapêutico , Antígenos E da Hepatite B/análise , Hepatite B Crônica/tratamento farmacológico , Hepatite B Crônica/imunologia , Interferon-alfa/uso terapêutico , Ribavirina/uso terapêutico , Adulto , Alanina Transaminase/sangue , Antivirais/efeitos adversos , DNA Viral/sangue , Quimioterapia Combinada , Feminino , Vírus da Hepatite B/genética , Hepatite B Crônica/sangue , Hepatite B Crônica/patologia , Humanos , Interferon alfa-2 , Interferon-alfa/efeitos adversos , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Mutação , Projetos Piloto , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/uso terapêutico , Retratamento , Ribavirina/efeitos adversos , Resultado do TratamentoRESUMO
The existence of four GB C virus/hepatitis G virus (GBV-C/HGV) subtypes has been reported. The subtype was determined in 16 multitransfused GBV-C/HGV infected patients prior to bone marrow transplantation by comparing the 5' untranslated region (5' UTR) sequence with 39 available sequences. Phylogenetic and bootstrap analyses were carried out with PHYLIP package 3.5c. In the samples with undefined subtype, the whole 5' UTR was cloned and sequenced. Comparison of distances showed that the isolates from 12/16 and 4/16 patients belonged theoretically to subtypes 2a and 2b, respectively. The phylogenetic tree and bootstrap analyses confirmed this result in only 11/16 samples. Analysis of the entire 5' UTR from the remaining five samples with undefined GBV-C/HGV subtype revealed genomic variability within the isolates from each patient and between the isolates of different patients. Evolutionary distances, phylogenetic tree, and bootstrap showed that the isolates from these samples were grouped in a separate branch, different from the published subtypes. In conclusion, a novel GBV-C/HGV subtype was found in a group of multitransfused patients with GBV-C/HGV infection.
Assuntos
Flaviviridae/isolamento & purificação , Neoplasias Hematológicas/virologia , Regiões 5' não Traduzidas/análise , Adulto , Evolução Molecular , Feminino , Flaviviridae/classificação , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , RNA Viral/análise , RNA Viral/sangue , Reação TransfusionalRESUMO
Serum samples from 20 anti-hepatitis B e antigen-positive patients with and without normal alanine aminotransferase (ALT) levels who had serum hepatitis B virus (HBV) DNA detectable only by polymerase chain reaction (PCR) were examined. Viral DNA was amplified by PCR, using primers that encompassed precore and ORF-X regions and sequenced directly, to investigate whether mutations in the nucleotide sequences of X and precore gene regions of HBV-DNA might be responsible for the difference in the activity of disease and in the levels of viral replication. The HBV-DNA concentration in patients with abnormal ALT levels was higher than in those with normal ALT. The amount of HBV-DNA correlated with the ALT levels (P < 0.05). Seventy-two percent of patients had HBV-DNA harboring the 1896 precore stop mutation, and there was a negative correlation between the percentage of precore mutant genotype and the HBV-DNA concentration (P < 0.05). Thirty percent of patients had mutations in ORF-X. Patients with ORF-X mutations had lower levels of HBV-DNA than those who had wild-type virus. The presence of mutations in precore and X regions may be related to a low HBV-DNA concentration and reduced biochemical activity in patients with anti-HBe.
Assuntos
Alanina Transaminase/sangue , Anticorpos Anti-Hepatite B/sangue , Antígenos E da Hepatite B/imunologia , Vírus da Hepatite B/genética , Hepatite B Crônica/virologia , Proteínas do Core Viral/genética , Adulto , DNA Viral/análise , Feminino , Genes Virais , Antígenos de Superfície da Hepatite B/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Fases de Leitura Aberta/genética , Reação em Cadeia da Polimerase/métodos , Transativadores/genética , Proteínas Virais Reguladoras e Acessórias , Replicação ViralRESUMO
The nucleotide sequence of hepatitis GB virus type C (HGBV-C)/hepatitis G virus (HGV) NS3/helicase and 5'-untranslated regions from 23 Spanish patients were analyzed to assign the HGV isolates one of the proposed HGBV-C/HGV genotypes. The analysis of the evolutionary distance frequency showed that the distances among all sequences in NS3/helicase region were distributed around a single peak of 0.20, suggesting that all included sequences belonged to the same HGBV-C/HGV genotype. By contrast, in the 5'-untranslated region, all the distances corresponding to our sequences and those of the HGBV-C/HGV types 2 and 3 were distributed around a major peak of 0.03. The remaining distances corresponding to the HGBV-C/HGV type 1 sequences were distributed around a minor peak of 0.11. The phylogenetic tree and pairwise comparison of evolutionary distances among the 5'-untranslated region of the infected patients and each HGBV-C/HGV genotype demonstrated that our HGBV-C/HGV isolates belonged to subtype 2a (17/23; 78%) and 2b (5/23; 22%). No relation was found between HGBV-C/HGV subtype and hepatitis B or C virus infection.
Assuntos
Flaviviridae/classificação , Hepatite B Crônica/virologia , Hepatite C Crônica/virologia , Filogenia , Regiões 5' não Traduzidas/genética , Adolescente , Adulto , DNA Viral/análise , Feminino , Flaviviridae/genética , Hepatite Viral Humana/virologia , Humanos , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , RNA Viral/sangue , Análise de Sequência de DNA , Espanha , Proteínas não Estruturais Virais/genéticaRESUMO
Fifty percent of healthy hepatitis B surface antigen carriers may have histologically proven chronic hepatitis. Our aim was to study the benefit of interferon-alpha in healthy patients. Twenty-nine hepatitis B surface antigen carriers with normal liver enzymes and with serum hepatitis B virus DNA were randomized into two groups: Group I, 14 patients treated with 9 megaunits of interferon alpha-2a thrice weekly for six months, and Group II, 15 control patients. A liver biopsy was obtained from each patient at study initiation. A second biopsy was available in nine treated patients and six controls. During treatment, a significant increase in alanine amino transferase levels was observed in treated patients as compared with the controls (P < 0.05). After treatment, transaminase levels decreased to normal values. No differences between treated and control patients were observed in clearance of hepatitis B virus markers. A significant increase in the total histological activity index between base line and final liver biopsies was observed in treated patients (P < 0.05). It is concluded that interferon alpha treatment may induce a biochemical and histological activation of liver disease. Accordingly, interferon alpha should not be administered to healthy hepatitis B surface antigen carriers, at least with the schedule used in this work.
Assuntos
Portador Sadio/terapia , Hepatite B/terapia , Interferon-alfa/efeitos adversos , Adulto , Alanina Transaminase/sangue , Portador Sadio/enzimologia , Portador Sadio/patologia , DNA Viral/sangue , Feminino , Hepatite B/enzimologia , Hepatite B/patologia , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Hepatite Crônica/enzimologia , Hepatite Crônica/patologia , Hepatite Crônica/terapia , Humanos , Interferon alfa-2 , Interferon-alfa/administração & dosagem , Fígado/enzimologia , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Proteínas RecombinantesRESUMO
The aim of this work was to study the presence of the hepatitis GB virus type C (HGBV-C) in liver and serum samples of children with chronic viral hepatitis, the time course of changes in viral RNA, and the possible acquisition routes of infection. Frozen serum and liver samples from 58 children with chronic hepatitis B (n = 33) or C (n = 25) were analyzed using polymerase chain reaction. Twenty-seven children had been included in different interferon trials. Two additional serum samples from the HGBV-C-positive children as well as serum samples from 29 of their relatives were also analyzed. HGBV-C RNA was detected in serum and liver samples from 9 of 58 (15%) of children as well as in serum samples from 3 of 29 of the relatives of the HGBV-C-infected children: the mother and the brother of one child (index case A) and the mother of another child (index case B). The homologies of the HGBV-C RNA sequences were 93% between index case A and his mother, 88% between index case A and his brother, and 94% between index case B and his mother. In the 3 children receiving alpha-interferon, HGBV-C RNA became undetectable during treatment although it reappeared in 2 of them after therapy. In conclusion, we found that 15% of children with chronic viral hepatitis were coinfected with HGBV-C. HGBV-C RNA was simultaneously present in serum and liver samples and tended to remain detectable even after alpha-interferon therapy. Our results suggest that vertical transmission of HGBV-C may occur.
Assuntos
Flaviviridae/isolamento & purificação , Hepatite B/virologia , Hepatite C/virologia , Fígado/virologia , RNA Viral/sangue , Adolescente , Adulto , Criança , Feminino , Flaviviridae/genética , Hepatite B/sangue , Hepatite B/transmissão , Hepatite C/sangue , Hepatite C/transmissão , Humanos , Transmissão Vertical de Doenças Infecciosas , Fígado/química , Masculino , Sondas de Oligonucleotídeos , Reação em Cadeia da PolimeraseRESUMO
We compared the response and the relapse rates of HBeAb-positive patients treated with interferon-alpha for 6 or 12 months. Thirty-eight HBeAb-positive patients which chronic hepatitis B were randomly allocated into two groups: Group I (19 patients receiving 10 MU of recombinant interferon-alpha 2b three times a week for 2 months, followed by 5 MU three times a week for 2 months and then 3 MU three times a week for 2 months); Group II (19 patients receiving 10 MU of recombinant interferon-alpha 2b three times a week for 2 months, followed by 5 MU three times a week for 2 months and then 3 MU three times a week for 8 months). At the end of treatment, alanine aminotransferase normalization was higher but not more significant in Group I than in II (53% vs 26%), while hepatitis B virus DNA clearance was similar in both groups (21% in Group I vs 26% in Group II). However, at 12 months of follow-up, biochemical relapses occurred only in Group I (60% vs 0% in Groups I and II, respectively). Five complete responders cleared hepatitis B surface antigen at that time. In conclusion, prolonged treatment of HBeAb patients is efficient in reducing the biochemical relapse.