Lectins as antimicrobial agents.

The resistance of micro-organisms to antimicrobial agents has been a challenge to treat animal and human infections, and for environmental control. Lectins are natural proteins and some are potent antimicrobials through binding to carbohydrates on microbial surfaces. Oligomerization state of lectins can influence their biological activity and maximum binding capacity; the association among lectin polypeptide chains can alter the carbohydrate-lectin binding dissociation rate constants. Antimicrobial mechanisms of lectins include the pore formation ability, followed by changes in the cell permeability and latter, indicates interactions with the bacterial cell wall components. In addition, the antifungal activity of lectins is associated with the chitin-binding property, resulting in the disintegration of the cell wall or the arrest of de novo synthesis from the cell wall during fungal development or division. Quorum sensing is a cell-to-cell communication process that allows interspecies and interkingdom signalling which coordinate virulence genes; antiquorum-sensing therapies are described for animal and plant lectins. This review article, among other approaches, evaluates lectins as antimicrobials.

[Autoimmune cholangiopathy]. / Colangiopatia auto-imune.

We report two patients with mixed characteristics of primary biliary cirrhosis and autoimmune hepatitis. They are two female patients in their fifties (54 and 58 years-old) one of them with autoimmune phenomenon, rheumatoid arthritis and Sjögren syndrome. Both of them showed laboratory values of cholestasis and the liver biopsy revealed liver cirrhosis with significant lesions of the bile ducts. They were treated with prednisolone with a rapid improvement and normalisation of their blood tests. They belong to a group called as overlap syndrome or autoimmune cholangitis with mixed characteristics, clinical, biochemical, immunological, and histopathologic overlapping between primary biliary cirrhosis and autoimmune hepatitis type I.

Alpha-interferon improves liver fibrosis in chronic hepatitis C: clinical significance of the serum N-terminal propeptide of procollagen type III.

Our objective was to estimate the effect of interferon (IFN) on the evolution of fibrosis in chronic hepatitis C and the significance of the N-terminal propeptide of procollagen type III (PIIIP) as a marker of fibrogenesis. One hundred seventeen patients, 72 male (61%) and 45 female (39%), with a mean age of 40.7+/-11.9 years were treated with a2b-IFN, 3 to 5 MU, for 12 months: sustained responders (SR = 44), relapsers (RR = 35), and nonresponders (NR = 38). Liver biopsies were performed before treatment and 1 year after cessation of IFN for evaluation of the histological activity index (HAI). Serum PIIIP was obtained at the time of liver biopsy, at the beginning, during, and end of therapy and during the follow-up. The normal value in 29 healthy individuals was 0.37+/-0.18 U/L. Staging was reduced in 58% of SR, 12.5% of RR, and 11.5% of NR. There was a correlation between PIIIP and the HAI before (n = 71, r(s) = 0.41, P < 0.0004) and after IFN (n = 71, r(s) = 0.58, P < 0.0001). The SR had a better improvement in grading (90.3%; P < 0.05) and staging (58%; P < 0.001). The correlation of the HAI parameters with the variation of PIIIP showed significance only for fibrosis (r(s) = 0.36, P < 0.002) and portal inflammation (r(s) = 0.35, P < 0.01). PIIIP normalized only in patients whose fibrosis improved (P < 0.01). At the end of therapy, PIIIP had a predictive value in the distinction of SR from RR (PPV, 64; PNV, 55.6). During the follow-up, PIIIP remained lower in SR compared with RR and NR (P < 0.002). The response to a-IFN improved liver inflammation and fibrosis. Serum PIIIP is a useful noninvasive method to evaluate serially fibrogenesis in chronic hepatitis C treated with IFN.

Hepatic stellate cell activation occurs in nonalcoholic steatohepatitis.

BACKGROUND/AIMS: Hepatic stellate cell activation has a major role in the pathogenesis of hepatic fibrosis, considered to constitute part of the healing response to a necroinflammatory stimulus. However, steatosis per se, has also been shown to induce this activation. This study evaluates if hepatic stellate cell activation is present, and how it correlates with steatosis, in nonalcoholic steatohepatitis, whose hallmark is steatosis. METHODOLOGY: Steatosis, hepatocyte damage, inflammation and fibrosis were graded from 0 to 3+, in liver biopsies from 15 well documented nonalcoholic steatohepatitis and 5 normal controls. Activated hepatic stellate cell activation were identified immunohistochemically using a monoclonal antibody raised against cytoplasmic alpha-smooth muscle actin, and semiquantitatively graded using a scoring method. RESULTS: Nonalcoholic steatohepatitis patients showed significantly greater numbers of alpha-smooth muscle actin-reactive hepatic stellate cell than controls: hepatic stellate cell index of 3.6 +/- 1.9 versus 1.5 +/- 0.5, P < 0.05. The distribution of alpha-smooth muscle actin-reactive hepatic stellate cell was higher in the perivenular areas, than in the intermediate zone and portal area, with no significant association between steatosis and alpha-smooth muscle actin-expressing hepatic stellate cell. However, a significant association was found between portal and lobular inflammation and hepatic stellate cell index, r = 0.72, P = 0.0005 and r = 0.75, P = 0.0002, respectively. CONCLUSIONS: This study demonstrates that hepatic stellate cell activation occurs in nonalcoholic steatohepatitis, clearly correlating with portal and lobular inflammation, but not with steatosis, suggesting that the mechanisms implicated in fibrosis in nonalcoholic steatohepatitis are probably related with inflammation.

Apoptotic cell death does not parallel other indicators of liver damage in chronic hepatitis C patients.

The mechanisms of hepatocyte damage and the events that lead to high rates of chronic liver disease in hepatitis C virus (HCV) infection remain unclear. Recent in vitro studies have suggested that the HCV core protein may disrupt specific signalling pathways of apoptosis. This prompted us to study patients with chronic HCV infection to: determine the extent of apoptosis in the liver; evaluate whether clinical and biochemical data are correlated with histological findings; and to investigate if apoptosis is related to the histological activity of the disease. Twelve patients with chronic hepatitis C were included in the study. Liver histology was scored by using the histological activity index (HAI) of Knodell et al. DNA fragmentation was assessed in liver tissue by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labelling (TUNEL) assay. Routine methods were used to determine serum markers of liver disease. Bile acids were measured in serum and liver by gas chromatography. Patients were placed, according to their HAI score, into group A (3.8 +/- 0.3) or group B (7.8 +/- 0.8) (P < 0.01). Liver enzymes tended to be higher in group B patients than in patients of group A. Levels of toxic bile acids in serum were greater in patients than in controls (P < 0.01). Chenodeoxycholic acid values were slightly higher in serum and liver of patients in group A. Liver biopsies with low HAI scores showed an increased rate of apoptosis (18.0 +/- 4.0 apoptotic cells per field) compared to those with higher HAI scores (6.6 +/- 2.1, P < 0.05) or to controls (3.5 +/- 0.4, P < 0.01). Hence, less severe liver disease, associated with lower histological grades and biochemistries, as well as increased levels of chenodeoxycholic acid, induces an expanded apoptotic response. The lower apoptotic rate in advanced liver disease may be associated with the high incidence of hepatocellular dysplasia/neoplasia.

Correlation of genotypes and route of transmission with histologic activity and disease stage in chronic hepatitis C.

Our objective was to evaluate the histopathological features of chronic hepatitis C of 64 liver biopsies and to correlate this with the route of transmission of hepatitis C virus, the genotype of HCV, and the patient's age. Moderate chronic hepatitis was the most frequently observed (62.5%). Cirrhosis was observed in 14 patients (21.9%) and was more frequently found among patients over 40 years of age (34.3% vs. 6.9%, P = 0.025). The mean histopathological activity index (HAI) was significantly higher in the sporadic (10+/-3.1) than the posttransfusional (7.5+/-3.7) and the intravenous drug use (IVDU) groups (6.3+/-2.8) (P<0.02). Moreover the sporadic group showed more fibrosis (P<0.04) than the posttransfusional group. No liver cirrhosis was found in the IVDU group. The overall prevalence of HCV variants was: 54.7% type 1b, 4.6% type 1a, 37.5% type 2c, 1.6% type 2b, 1.6% type 2. The genotype distribution showed no relation to the HAI, hepatitis activity (grade), and fibrosis (stage) of the liver disease. In conclusion, the sporadic route of transmission of HCV was related to a more severe chronic hepatic disease, a finding that could influence future antiviral therapies. The predominance of HCV type 1b in this study reflects the higher frequency of this variant in our area. Our data suggests that the ultimate consequence of HCV chronic infection depends on patient age rather than on HCV genotype.

Serum gastrin and gastrin-immunoreactive cells in the antral mucosa of patients with alcoholic liver disease.

BACKGROUND/AIMS: Hypergastrinaemia has been reported in liver cirrhosis; meanwhile, it is unclear whether it is associated with an increase in gastrin cell function. The serum gastrin concentration and the number of gastrin cells in antral biopsies were studied in patients with alcoholic liver disease. METHODS: Immunocytochemical and quantification techniques were used to localize and determine the number of gastrin cells. RESULTS: Slight non-significantly higher serum gastrin values were observed in the alcoholic liver disease patients compared with controls, but the individual variation within the groups was considerable. The frequency of gastrin cells did not differ between groups. However, the size of the gastrin cell nuclei was larger in patients with liver disease than in controls, indicating increased cellular activity. CONCLUSIONS: Alcoholic liver disease, with a disturbed liver function, influences the gastrin cells. The observed alterations may reflect the effect of alcohol and/or malnutrition, or may be secondary to the influence of liver disease on other regulatory peptides.

Long-term gallbladder stone recurrence and risk factors after successful lithotripsy.

OBJECTIVE: The risk of recurrence has limited the acceptability of conservative therapies of gallbladder stones. The aim of the present study was to determine the rate of stone recurrence and its risk factors, after successful shock-wave lithotripsy. DESIGN: Prospective ultrasound follow-up at yearly intervals or whenever biliary pain was reported. METHODS: One hundred and fifty-eight consecutive patients (single stone, n = 130; two or three stones, n = 28) were followed up to 70 months (median, 33 months) after stone disappearance and discontinuation of ursodeoxycholic acid. RESULTS: Forty-three patients developed recurrent stones. By actuarial analysis, the recurrence rates (as a percent) at 1, 2, 3, 4 and 5 years were, respectively: 6+/-2, 14+/-3, 27+/-4, 35+/-5, and 44+/-6 (observed +/- SE). Cox's regression analysis identified high body mass indexes to be a risk factor (P = 0.02) for newly formed stones. Having had a single primary stone did not seem to be protective. Fourteen of the 43 patients (33%) had early symptoms of recurrence. Thirty-eight patients (89%) chose to undergo oral dissolution again, which was complemented by lithotripsy in eight patients (19%). CONCLUSION: The 5-year gallbladder stone recurrence after lithotripsy and ursodeoxycholic acid is not substantially smaller than that reported by post-bile acid studies. Obesity is a risk factor for recurrent stones as it is for primary stones. Most patients with secondary stones choose to have conservative therapy again. Gallbladder stone recurrence still is one of the major drawbacks of these treatments and cost-effective strategies are needed to prevent it.

Autoimmune cholangitis: case report.

We report on 2 patients who showed mixed signs of primary biliary cirrhosis and autoimmune hepatitis. Both patients were female, in their fifties (54 and 58), their laboratory tests indicated cholestasis, and a liver biopsy revealed liver cirrhosis with significant lesions of the bile ducts. Both were treated with prednisolone with their liver tests showing a rapid normalization of their aminotransferases. These patients can be considered as presenting with what is known as the overlap syndrome or autoimmune cholangitis, which has the clinical, biochemical, immunological, and histopathological characteristics of primary biliary cirrhosis and autoimmune hepatitis type I.

Non-alcoholic fatty liver: another feature of the metabolic syndrome?

BACKGROUND AND AIMS: Hepatic steatosis and nonalcoholic steatohepatitis (NASH) have been associated with obesity, non insulin-dependent diabetes mellitus and hyperlipidemia. The present study was designed in order to evaluate whether patients with steatosis/NASH presented common features with the metabolic syndrome. METHODS: In 30 patients with nonalcoholic fatty liver the prevalence of hypertension and diabetes; the glucose/insulin profile, lipid profile, and serum leptin were evaluated and correlated with body composition and energy expenditure, assessed by bioimpedance spectroscopy and indirect calorimetry, respectively. Results were compared with a group of eight controls. RESULTS: Obesity was present in 80% of patients, hypertension in 50% and non insulin dependent diabetes in 33%. Glucose metabolism was altered in 69%, with elevated insulin in 14 patients. Serum leptin, higher in women, was increased in patients: 33.9 +/- 38.9 vs 9.6 +/- 6.9 ng/ml, P< 0.05. There was a correlation between insulin and leptin, both of which correlated with body mass index, fat mass and percentage of body fat. Dyslipidaemia was found in 80% of patients: 45% presented low high density lipoproteins cholesterol, 58% high low density lipoproteins and 38% elevated very low density lipoproteins. CONCLUSIONS: There is a strong association between nonalcoholic fatty liver and features of the metabolic syndrome, suggesting a simultaneous insulin resistance and decreased sensitivity to leptin.

Delayed clearance of serum HBsAg in compensated cirrhosis B: relation to interferon alpha therapy and disease prognosis. European Concerted Action on Viral Hepatitis (EUROHEP)

OBJECTIVE: The aim of this study was to evaluate the incidence, prognostic factors and clinical significance of delayed clearance of serum HBsAg in compensated cirrhosis B. METHODS: This was a retrospective cohort study of 309 consecutive white patients with biopsy-proved compensated cirrhosis type B. RESULTS: During a mean follow-up of 68 months, HBsAg loss occurred in 32 patients, including 16 (8%) of 196 untreated patients (mean annual incidence 0.8%), 8 (10%) of 82 interferon (IFN) alpha-treated patients and eight patients who had been treated with other antivirals or steroids. The 5-yr probability of HBsAg loss was 4% and 16% for untreated and IFN-treated patients, respectively (p = 0.0001). Cox's regression analysis identified hepatitis B e antigen-positivity at entry as the sole independent prognostic factor for HBsAg loss. Of the 32 patients who lost HBsAg, one (3%) subsequently developed hepatocellular carcinoma (HCC) and died, whereas, among the patients who remained HBsAg-positive, 11% developed HCC and 20% had died. The probability of HCC appearance was lower (p = 0.0137) and survival was longer (p = 0.0006) in patients who cleared HBsAg compared with patients with HBsAg persistence. CONCLUSION: The incidence of HBsAg loss is about 0.8% in cirrhosis type B. Prognostic factors for clearance of HBsAg are initial HBeAg positivity and therapy with alpha interferon. Patients with cirrhosis type B, who lose HBsAg, have a low risk for liver cancer or liver-related death.

Diphenylhydantoin-induced hepatitis: a case report.

This case report is of a 25-year old man with severe diphenylhydantoin-induced hepatitis, fever, rash and generalized lymphadenopathy. Hepatic histology and lymphocyte transformation tests strongly supported the diagnosis. He made an uneventful recovery with supportive therapy.

[Vaccine against hepatitis B. Eight years of experience]. / Vacinação contra a hepatite B. Oito anos de experiência.

Hepatitis B virus infection is one of the world's major health problems and health care workers are especially at risk, leading to the need for a high priority vaccination program. Such a program was begun in 1989 in S. Maria Hospital and included all hospital personnel and students of the Faculty of Medicine. The screening included 2360 health care workers and 1153 students. Fifty-five percent of hospital health care workers and 41% of the students participated in the vaccination program. The overall prevalence of hepatitis B markers was 16.8%, for hospital personnel, the chronic carrier being 0.95%, and for the students, 5.5% and 0.3% respectively. The highest prevalence was observed in the Biochemical Laboratory--64% (7/11), Surgery--42% (13/31), Pneumology--39% (9/23), Emergency Department--29% (7/24), Hematology Laboratory--29% (7/24) and Orthopedics--29% (10/35). The prevalence was also higher in students in the last three years of Medical School compared to those in the first three years, 12.2% vs 7.2%, p = NS. Local pain has been the most frequent complaint in 8.6% of vaccinees. The control with the quantification of anti-HBs in the 7th month has shown a serological efficacy of about 95%. A non-response was observed in male workers, 13% compared to 5% for females, p < 0.05. Older employees also showed higher non-response: the average age of workers with anti-HBs of 0 U/L was 52.3 years, and those with anti-HBs of more than 100 U/L was 38.4 years, p < 0.02. Hepatitis B vaccine is a safe and effective preventative measure that has been widely used for years. Our study shows the need for a more aggressive approach to the vaccination of health care workers because a significant percentage of them in our country are not protected.

The changing epidemiological pattern of hepatitis A in Lisbon, Portugal.

OBJECTIVE: To evaluate the prevalence of anti-hepatitis A virus (HAV) in an urban population, in order to assess the changing epidemiological pattern of hepatitis A. SUBJECTS: We studied 325 health care workers from Santa Maria Hospital and 201 students of the Medical School of Lisbon. RESULTS: The prevalence of anti-HAV was lower in the younger age groups: 29% in the first 3 years of Faculty compared to 46% in the last 3 years, P < 0.001. The overall prevalence for students was 35%, whereas in health care workers of less than 30 years it was 65%, P < 0.001. These findings show a declining prevalence of anti-HAV, particularly in younger age groups, when compared with the results obtained in 1983, which showed a prevalence of 85%. CONCLUSION: The epidemiological pattern of hepatitis A in Lisbon, Portugal, is changing in some urban groups, with prevalences approaching those of more developed countries.

Centrocytic lymphoma presenting as a subphrenic abscess and a solitary liver nodule.

Non Hodgkin's lymphoma revealed by hepatic manifestations is extremely rare. We describe here a 82-year old male patient who presented with a right subphrenic abscess and a solitary liver tumour that was shown to be a centrocytic lymphoma. Furthermore, asymptomatic cryptogenic liver cirrhosis was diagnosed. This previously unreported form of clinical presentation of a non Hodgkin's lymphoma as well as the association with liver cirrhosis are discussed in the context of the recent literature.

Streptococcus bovis spontaneous bacterial peritonitis in patients with alcoholic cirrhosis.

Spontaneous bacterial peritonitis (SBP) is a frequent cause of decompensated alcoholic cirrhosis. The authors describe the first two cases caused by infection with Streptococcus bovis. They suggest that this microorganism may be present in the intestinal flora of these patients more frequently than assumed. Furthermore, the course of SBP due to S. bovis may be particularly severe.

Nonalcoholic steatohepatitis. Clinicopathological comparison with alcoholic hepatitis in ambulatory and hospitalized patients.

This study reports a clinicopathological analysis of 105 patients whose liver histology showed a pattern of alcohol-like steatohepatitis. There were 32 nonalcoholic, 21 asymptomatic ambulatory, and 52 hospitalized alcoholic hepatitis patients. Female sex, obesity, and diabetes predominated in nonalcoholics. Clinical and laboratory presentation were similar in nonalcoholics and ambulatory alcoholics, but different from the hospitalized alcoholics. Histology showed an increasing degree of severity of hepatocellular damage, Mallory bodies, neutrophil and mononuclear infiltration, and pericellular and portal fibrosis from the nonalcoholics to the hospitalized alcoholics, with ambulatory alcoholics displaying an intermediate degree of severity. Steatosis and glycogenated nuclei were prevalent in the obese, diabetic nonalcoholics, of whom 47% had significant fibrosis and 8% cirrhosis, the latter present in 38% and 89% of ambulatory and hospitalized alcoholic hepatitis (P = 0.0001), respectively. In asymptomatic subjects with suspected liver disease, a liver biopsy is the only way of establishing the type and severity of liver lesions.

Tamoxifen-associated steatohepatitis--report of three cases.

The authors describe three cases of tamoxifen-associated steatohepatitis, which resulted from a daily dosage of 20 mg used as the adjuvant treatment of breast carcinoma. Liver tests became normal after discontinuation of tamoxifen.

An aggressive protocol of ESWL and dissolution therapy of gallbladder stones.

OBJECTIVES: to assess the overall efficacy and which factors are independent predictors of success of ESWL and oral dissolution therapy of gallbladder stones using an aggressive protocol (high energy shock waves -median 22 Kv- and allowance to up to 6 sessions with an electro-hydraulic lithotripter). PATIENTS AND METHODS: inclusion criteria were 1) biliary pain; 2) 1 to 3 radiolucent stones or with slight calcification; 3) total stone volume under 15 cm3, equivalent to a single stone 3 cm diameter and 4) opacified cholecystography. Data was collected prospectively for 139 consecutive patients undergoing this treatment and the stone-free curves up to 12 months were analyzed as a function of age, sex, body-mass index, total stone volume, number of stones and the presence of slight calcification. RESULTS: patients underwent a mean of 2.6 sessions (range: 1-6) and 2834 shock waves (range: 589-8175). The global stone-free rate at 12 months was 54% (95% confidence interval: 45-64%). Factors that significantly -and adversely- influenced outcome were total stone volume (P < 0.001), number of stones (P = 0.005) and slight calcification (P = 0.038), using Cox's regression. Beyond significance, these three factors showed a marked effect on the stone-free curves. CONCLUSIONS: our data suggest that, even with this aggressive protocol, these factors are clearly detrimental. Thus, the results of our study agree with the current trend to restrict this combination therapy to patients with single, non-calcified stones with a small volume, or up to 2 cm diameter as is usually quoted.

[Echo endoscopy in the local staging of gastric cancer]. / A eco-endoscopia no estadiamento local do cancro gástrico.

OBJECTIVES: to evaluate the accuracy of endoscopic ultrasonography (EUS) in the pre-operative T and N staging of gastric cancer. PATIENTS AND METHODS: 41 consecutive patients with gastric cancer (35 carcinomas and 6 lymphomas) underwent EUS using an Olympus GF-UM20, with 360 degrees sector scan and interchangeable frequency (7.5-12 MHz). They were classified as T1-T4 and N0-N2, according to the TNM system. These results were then compared with the surgical and pathological staging (SP), by the weighted K statistic (Kw). RESULTS: In 2 patients EUS staging was not possible due to cardiac stenosis and in other 4 patients, information about SP staging was not available. In the remaining 35 patients the EUS/SP agreement was good (Kw = 0.80) for stage T and moderate (Kw = 0.49) for stage N. Non-agreement was mostly due to EUS understaging, both T and N. CONCLUSIONS: 1) EUS is the most accurate procedure for pre-operative local staging of gastric cancer, and 2) EUS should preferably be performed by endoscopists with sonographic experience and a particular interest in the method.