Virologic response and resistance to lamivudine in patients with chronic hepatitis B: a ten-year retrospective analysis.

BACKGROUND: Emergence of resistance was recognized shortly after the introduction of lamivudine. This 10 year retrospective study investigates resistance to lamivudine and the modifications of antiviral strategies required. PATIENTS AND METHODS: Two hundred and nine patients were treated with lamivudine. Sixty seven out of 209 patients were excluded from the present study. HBVDNA was tested using the PCR assay and genotypic resistance was performed using the direct PCR sequencing. RESULTS: In the 125 patients initially treated with lamivudine monotherapy: Α) 48 (38.4%) patients with a mean time of 63.6±26.2 months under lamivudine treatment have normal ALT levels with negative (19%) or low (<1X102) HBVDNA levels, 10% developed cirrhosis, 1 HCC and 6% cleared HBsAg. Β) Resistance was developed in 61.60% patients within 45±23.84 months of lamivudine treatment. These patients were: 1) either switched to adefovir (9), entecavir (2) or tenofovir (2) or adefovir was added to lamivudine (21) for a short time and then they were switched to adefovir alone. Six out of 34 patients developed cirrhosis and 4 HCC while on treatment. 2) or adefovir was added-on to lamivudine (43). In 39 out of 43 treatment is ongoing while on virological response. No one developed cirrhosis or HCC. C) Seventeen patients received de novo combination therapy with lamivudine and adefovir and 2 out of 17 (11.7%) showed resistance to adefovir after 24 months of therapy. CONCLUSIONS: Our results showed that a) approximately 38.4% of patients maintain viral suppression more than 5 years of lamivudine treatment and b) rescue therapy with add-on adefovir to ongoing lamivudine, seems to be a better treatment strategy associated with long term benefit regarding disease complications.

Health-related quality of life in Greek chronic hepatitis C patients during pegylated interferon and ribavirin treatment.

UNLABELLED: Background - Aims: Chronic hepatitis C (CHC) can cause a series of neuropsychiatric symptoms, whereas the currently approved treatment for this disease often induces similar symptoms as well. The aim of the present study was to compare Greek CHC patients' health-related quality of life (HRQoL) with that of healthy controls, to identify any possible relationships between HRQoL and demographic and laboratory parameters and to study the fluctuation of HRQoL during therapy and follow-up. PATIENTS AND METHODS: Ninty nine patients with CHC and 91 healthy controls were enrolled in the study. ALT, viral load, HCV genotype, fibrosis stage by liver biopsy and BMI, were determined at baseline. All patients completed the SF-36 quality of life questionnaire, which was self-administered, before treatment. They were treated with pegylated interferon alpha2-a or alpha-2b and ribavirin for 24 or 48 weeks and evaluated in the middle of therapy, at the end and six months after treatment cessation. SF-36 questionnaire was also completed in each evaluation. RESULTS: Patients' HRQoL was found to be below that of healthy controls in all SF-36 scales before treatment. There was a significant negative association between history of drug abuse and general health and a positive association between age and mental health. Multivariate analysis revealed that history of drug abuse seemed to play a significant role in bodily pain and general health of patients, as well as age did in vitality and mental health. The course of patients' HRQoL showed that in the middle of treatment values in all SF-36 scales were below those of baseline and they returned to pretreatment levels at the end of therapy. However, at the end of the six month follow-up period, an improvement in almost all scales compared to baseline was noted. CONCLUSION: Our results showed that a) Greek CHC patients' HRQoL was worse than that of healthy individuals and fluctuated significantly during treatment b) A history of drug abuse and age can independently affect HRQoL c) During treatment values of HRQoL are worsened possibly due to interferon-a treatment and d) In the long-term treatment results in improvement of HRQoL.

Endothelial nitric oxide synthase (eNOS) is not upregulated in gastric mucosa of Helicobacter pylori (H. pylori)-positive patients with type 2 diabetes mellitus.

AIM: To evaluate the expression of eNOS and CD34 in gastric mucosa of Helicobacter pylori (H. pylori) positive diabetic patients, in correlation with glycaemic control and diabetic autonomic neuropathy (DAN). METHODS: We prospectively studied 49 diabetic type 2 patients (29 women, mean age 65.32+/-8.56 years) and 30 control subjects (15 women, mean age 58.47+/-12.40) that underwent endoscopy. Biopsies from the body and antrum were evaluated for H. pylori-gastritis, eNOS and angiogenic marker CD34 expression. Statistical analysis in correlation with mean glycosylated haemoglobin (HbA1c) of the last 3 years, and DAN was performed. RESULTS: The two groups were matched for age (p=0.144), sex (p=0.335), H. pylori-infection (p=0.617) and degree of gastritis (p=0.78). eNOS and CD34 attenuated expression correlated with diabetes mellitus (DM) in the corpus (p=0.009 and 0.02, respectively). eNOS and CD34 expression was upregulated in H. pylori-positive controls but not in H. pylori-positive diabetic patients (p=0.010 and 0.007 for the corpus and p=0.036 and 0.047 for the antrum, respectively). eNOS expression correlated with good glycaemic control (GGC) in the gastric corpus (p<0.001) and antrum (p=0.0037) and with absence of DAN (p=0.009 and 0.036, respectively for the corpus and antrum). CONCLUSION: Chronic glycaemic control affects eNOS expression and angiogenesis in the gastric mucosa of patients with type 2 DM. eNOS expression is not upregulated in H. pylori-positive diabetic patients.

A short course of preoperative radiotherapy improves prognosis of operable rectal carcinoma: a case control study.

BACKGROUND/AIMS: Randomized Swedish studies demonstrate the efficacy of a 5-fraction course of preoperative radiotherapy for rectal carcinoma. The present study evaluates the results in a single Greek institution over a 10-year period, with a similar regimen. METHODOLOGY: During the period of 1995-2000, 150 consecutive patients with Dukes' B or C rectal cancer were matched to receive preoperative radiotherapy (Group I) or not (Group II). Seventy-five patients received pelvic radiotherapy of 2500cGY/5 fractions, followed by surgery within one week. Radiotherapy was delivered through 4 portals, with the patient lying in the prone position. A CT scan was used to define treatment volume. The 5-fraction course was used for lesions that seemed readily resectable. Patients in both groups received adjuvant chemotherapy. Local recurrence, disease-free interval and 5-year survival were evaluated and analyzed. RESULTS: The disease-free interval was significantly longer in Group I (p < 0.0005). This benefit was mainly due to a significantly lower incidence of local recurrence in Group I (9/75, 12%) compared with Group II (30/75, 40%) (p < 0.0005). The incidence of distant metastases was not significantly different between the 2 groups. The 5-year survival for all patients, who underwent "curative" surgery was significantly higher in Group I (77.3%) as compared to Group II (39%), (p < 0.0005). CONCLUSIONS: Patients with resectable rectal cancer who received 2500cGy/5 fractions preoperative radiotherapy to the pelvis had excellent local control of disease, longer disease-free interval and higher 5-year survival than patients who did not. These patients were able to undergo sphincter preserving surgery and adjuvant chemotherapy.

Cytokine mRNA expression in hepatitis C virus infection: TH1 predominance in patients with chronic hepatitis C and TH1-TH2 cytokine profile in subjects with self-limited disease.

Many determinants of the immune response have been implied in the pathogenesis of chronic hepatitis C. TH1 and TH2 cytokines play a prominent role in viral infections and a dysregulation of these cytokines could account for viral persistence and evolution of chronic disease. To explore a possible TH1 and TH2 cytokine dysregulation resulting in the inability to terminate hepatitis C virus (HCV) infection, we studied TH1 [interferon (IFN)-gamma, interleukin (IL)-2] and TH2 (IL-4, IL-10) mRNA expression of peripheral blood mononuclear cells (PBMC) in response to NS3 HCV antigen stimulation, in 31 untreated patients with chronic hepatitis C and 29 subjects with self-limited disease. After a 48 h culture of PBMC, total RNA isolation was performed and complementary DNA was prepared by reverse transcription. mRNA levels were quantified by real-time polymerase chain reaction using a standard curve formed after cloning each cytokine gene and a reference gene using recombinant DNA technology in a specific plasmid vector. In the patients group, mRNA expression of IFN-gamma, IL-2 and IL-4 but not IL-10 was detected, IFN-gamma being the predominant cytokine expressed. All four cytokines were expressed in subjects with self limited disease, however levels of IFN-gamma were lower and a significant higher expression of IL-10 compared to patients was found. There was a significant correlation between IFN-gamma mRNA expression levels and stage of fibrosis. Our findings show that in chronic hepatitis C, TH1 cytokines predominate and correlate to liver immunopathology. Furthermore, subjects with self-limited disease, maintain the ability to respond to HCV antigens for a long time after disease resolution.

Treatment of intravenous drug users with chronic hepatitis C: treatment response, compliance and side effects.

BACKGROUND: Although intravenous drug users (IVDUs) comprise the majority of patients with chronic hepatitis C, most of them are excluded from treatment because of concerns about adherence to treatment and side effects. MATERIAL AND METHODS: In this study we retrospectively evaluated safety, compliance to treatment and efficacy of treatment in IVDUs with HCV infection in 163 former IVDUs with chronic hepatitis C, who were not in methadone substitution and were attending our clinics the period 1997-2004. All subjects were HCVRNA (+), had ALT levels>x1.5 UNL and were treated for their HCV infection. Treatment consisted of three different regimens: IFN-alpha monotherapy (39.8%), IFN-alpha/ribavirin combination therapy (30.1%) and pegylated IFN-alpha/ribavirin combination therapy. RESULTS: Eighty seven over 163 patients (53.3%) discontinued treatment early due to drug abuse relapse (62%), side effects (32.1%, 10% psychiatric) and 5,7% for other reasons. Eighty precent of those who discontinued treatment had pre-treatment drug abstinence/= 9 months.

The effect of aspirin and high fibre diet on colorectal carcinoma: a comparative experimental study.

BACKGROUND: The aim of this study is to assess the role of high fibre diet and aspirin on dimethylhydrazine (DMH)-induced colorectal cancer in rats. MATERIALS AND METHODS: Colorectal tumours were induced with DMH. The animals were randomly divided into five groups (15 rats each): I, controls; II, rats receiving only the carcinogen; III, rats receiving the carcinogen and high fibre diet; IVA, rats receiving the carcinogen plus low dose aspirin; IVB, rats receiving the carcinogen plus high dose aspirin. RESULTS: Adenocarcinomas were detected in 100% of the rats in group II, 47% of the rats in group III (chi2, p<0.05), 100% of the rats in group IVA, but the incidence was reduced to 50% in the rats of group IVB (p<0.05). CONCLUSIONS: These data indicate that high fibre diet and aspirin suppress experimental colon carcinogenesis and the protective effect of aspirin is dose related.

Angiogenesis and apoptosis-related protein (p53, bcl-2, and bax) expression versus response of gastric adenocarcinomas to paclitaxel and carboplatin chemotherapy.

The role of angiogenesis and apoptosis-related proteins in defining response to chemotherapy is poorly understood. We examined the microvessel density (MVD) and the expression of p53, bcl-2, and bax proteins in a series of 28 locally advanced gastric adenocarcinomas, treated with paclitaxel and carboplatin. A strong cytoplasmic reactivity in more than 10% of cancer cells was recorded in 25% of cases for p53 protein, and in 14% and 64% of cases for bcl-2 and bax proteins, respectively. Microvessel density was assigned in three categories: low (<35), medium (35-60), and high (>60). Tumors of medium MVD showed a significantly higher response rate compared with those of high or low MVD (p = 0.01 and 0.001, respectively), and prognosis was significantly better in this group of patients with medium MVD tumors (p < 0.02). Loss of bax protein expression was somewhat more frequent in tumors resistant to chemotherapy, but this difference was not of statistical significance. Nuclear p53 reactivity was associated with higher MVD (p = 0.02). The expression of p53 and bcl-2 did not influence the outcome of treatment. The present study suggests that although apoptosis-related proteins may have a role in defining response to taxanes, parameters related to tumors' vasculature, such as drug availability or angiogenic tissue regeneration, may be equally important.

Bax protein expression in colorectal cancer: association with p53, bcl-2 and patterns of relapse.

This study evaluated the frequency and the prognostic significance of bax, bcl-2 and p53 proteins in stage B and C adenocarcinomas of the colon and rectum. Paraffin-embedded specimens from 268 patients with colorectal adenocarcinomas, treated with surgery, were assessed; of these 160 cases were Duke's stage B and 108 cases were Duke's stage C disease. Adjuvant chemotherapy was given to all stage C and to 108 out of 160 stage B cancer patients, while those having rectal malignancy also received pelvic radiotherapy. Duke's stage B patients were treated either with surgery alone or with surgery and radiotherapy. The follow-up period at the time of analysis ranged from 12-72 months (median 32 months). Immunohistochemical expression of bax, bcl-2 and mutant p53 proteins was detected with a frequency of 42%, 37% and 48%, respectively. However, the expression was strong only in 17% of tumours, on average. A strong bcl-2 expression was significantly associated with a strong bax expression (p < 0.0001) and with absence of p53 nuclear accumulation (p < 0.005). There was, however, no correlation between bax and p53 proteins. Furthermore, bcl-2 expression was significantly more frequent in grade I and 2 adenocarcinomas compared to grade 3 disease (p = 0.01). In stage B (but not C) adenocarcinomas, bax expression was directly associated with higher risk of local relapse (p = 0.04). By contrast, cases with p53 nuclear accumulation, when they had received adjuvant radiotherapy, were significantly associated with a lower incidence of local relapse (p = 0.01), but a higher rate of distant metastasis (p = 0.06). Multivariate analysis for disease free and overall survival showed that bax expression and high Duke's stage were independent prognostic parameters associated with an unfavourable outcome (p = 0.009 and p = 0.0001, respectively). It was concluded that the immunohistochemical expression of bax is a marker of poor prognosis and of a higher risk of local relapse in patients with colorectal adenocarcinomas. p53 nuclear accumulation is associated with a better local control, following radiotherapy and with a metastatic phenotype. The development of novel monoclonal antibodies recognising specifically the mutated versus the wild type form of proteins would apparently improve the prognostic and predictive value of the immunohistochemically detected apoptotic proteins.

Immunohistochemical phenotyping and PCNA detection in gastrointestinal stromal tumors.

The immunohistochemical profile and the expression of proliferating cell nuclear antigen (PCNA) were studied in a series of 44 mesenchymal tumors of the gastrointestinal tract (GIT). On routinely hematoxylin-eosin stained sections 31 cases were classified as leiomyomas or leiomyomatoid tumors, 12 as leiomyosarcomas and 1 as a neurilemmoma. Immunohistochemical stains for smooth muscle antigen (SMA), S-100 protein, glial fibrillary acidic protein (GFAP), vimentin and desmin were performed with the peroxidase-antiperoxidase method on paraffin sections. The streptavidin-biotin method for PCNA immunostaining was applied using the monoclonal antibody PC 10. On the basis of immunohistochemical findings, 32 cases were identified as leiomyomatoid tumors or leiomyosarcomas (SMA positive, S-100 protein negative), 2 cases as nerve sheath tumors (SMA negative, S-100 protein and GFAP positive), whereas 8 cases presented a mixed phenotype (SMA positive and S-100 protein positive). Two cases were negative for both SMA and S-100 protein. All tumors showed positive immunostaining for vimentin and a negative one for desmin. There was a correlation between the histologic grade and proliferating score, displayed by PCNA expression in tumors of smooth muscle origin. The PCNA expression in tumors of mixed phenotype was intermediate to that seen in leiomyosarcomas (high expression) and in leiomyomas (low expression).

Changes of hepatitis B core antigen (HBcAg) in liver biopsies of patients with chronic active hepatitis B treated with interferon.

Data have shown that hepatitis B core antigen (HBcAg) is detected in both the hepatocyte nucleus and cytoplasm. Its expression is associated with chronic hepatitis and active viral replication. The intrahepatic distribution of HBcAg was studied in liver biopsies of 14 patients with chronic active hepatitis B (CAH-B) (5 were hepatitis B e antigen [HBeAg]+/anti-HBe--, 9 were HBeAg--/anti-HBe+) by an immunohistochemical method (PAP) before and after 6-month treatment with interferon (IFN), and our findings were analyzed according to the response of patients to treatment. Our findings showed that, at the end of treatment, nuclear HBcAg was decreased or absent in 4 of 5 and cytoplasmic HBcAg in 2 of 4 HBeAg+/anti-HBe--patients, irrespective of the response to treatment. Loss of cytoplasmic expression was related to the outcome of treatment in 5 of 9 HBeAg--/anti-HBe+ patients. Four patients expressed no HBcAg before or at the end of treatment. These findings possibly reflect a different pattern of viral core antigen expression as a result of IFN therapy in the two groups of patients.

The value of neuroendocrine markers in non-small cell lung cancer: a comparative immunohistopathologic study.

In order to estimate the value of immunohistochemical identification of neuroendocrine (NE) differentiation markers in non-small cell lung carcinomas (NSCLCs), we investigated the expression of five neuroendocrine and neural differentiation-related antigens in 51 NSCLCs. Additionally, 20 epithelial lung tumors with NE differentiation [15 carcinoids and five small cell lung carcinomas (SCLCs)] and 61 epithelial tumors of various other origin (breast, prostate, colon and head-neck carcinomas) were studied. An indirect two-stage immunoperoxidase method was performed in formalin-fixed and paraffin-embedded tissue specimens, by using commercially available monoclonal antibodies. These antibodies are directed against neuron-specific enolase (NSE), chromogranin-A and Leu-7 which are general markers of NE differentiation, bombesin, which is a specific NE secretory product and neurofilament triplet protein (NFTP), an intermediate filament protein of neuronal differentiation. All five markers demonstrated a positive immunoreactivity in NSCLCs, equally distributed to all three histologic subtypes, ranging from 16 to 47% of the cases (NSE 47%, bombesin 21.5%, Leu-7 21.5%, chromogranin-A 18% and NFTP 16%). Most of the carcinoids and SCLCs expressed multiple or all NE markers. The other four epithelial tumors showed a positive immunoreactivity for bombesin, Leu-7 and NFTP, ranging from 11 to 40% of the cases. Chromogranin-A was not expressed in any of these tumors, whereas NSE was demonstrated only in 17% of breast carcinomas. The following remarks can be drawn from this study: (1) some NSCLCs showed immunophenotypic NE differentiation; (2) among all the markers used, NSE was the most sensitive (sensitivity, 100%) and chromogranin-A the most specific (specificity, 100%); and (3) NSE and chromogranin-A appear to be the most valuable and useful indicators of probable neuroendocrine differentiation in lung epithelial tumors.

Epstein-Barr virus and primary lung lymphoma: a study utilizing the polymerase chain reaction.

Seven cases of primary lung lymphoma were analyzed for the presence of Epstein-Barr virus DNA sequences by the polymerase chain reaction. The series included: four cases of diffuse, small lymphocytic lymphoma; one case of diffuse, intermediate lymphocytic lymphoma; one case of diffuse, small cleaved cell lymphoma; and one case of large cell, immunoblastic lymphoma. The latter occurred in a patient with acquired immunodeficiency syndrome. A 200-bp sequence of the Epstein-Barr nuclear antigen 1 gene was used as a template for PCR amplification. The only tumor that contained Epstein-Barr virus sequences was the immunoblastic lymphoma. These findings support previous observations that small lymphocytic lymphomas of the lung are not related to Epstein-Barr virus infection. In contrast, some large cell lymphomas may represent Epstein-Barr-virus--associated lymphoproliferative disorders.