Long-term outcome of chronic hepatitis B in Caucasian patients: mortality after 25 years.

OBJECTIVE: To assess risk factors for liver-related death, we re-evaluated, after a median follow-up of 25 years, a cohort of 70 Caucasian patients with hepatitis B e antigen (HBeAg) positive chronic hepatitis (CH) at presentation. METHODS: Follow-up studies included clinical and ultrasound examinations, biochemical and virological tests, and cause of death. RESULTS: Sixty-one (87%) patients underwent spontaneous HBeAg seroconversion. During a median period of 22.8 years after HBeAg seroclearance, 40 (66%) patients became inactive carriers, whereas the remaining 21 (34%) showed alanine aminotransferase elevation: one (1%) had HBeAg reversion, nine (15%) detectable serum HBV DNA but were negative for HBeAg, eight (13%) concurrent virus(es) infection and three (5%) concurrent non-alcoholic fatty liver disease. Liver-related death occurred in 11 (15.7%) patients, caused by hepatocellular carcinoma in five and liver failure in six. The 25-year survival probability was 40% in patients persistently HBeAg positive, 50% in patients with HBeAg negative CH or HBeAg reversion and 95% in inactive carriers. Older age, male sex, cirrhosis at entry and absence of sustained remission predicted liver-related death independently. The adjusted hazard ratios (95% CI) for liver related death were 33 (3.01-363) for persistently HBeAg positive patients and 38.73 (4.65-322) for those with HBeAg negative CH or HBeAg reversion relative to inactive carriers. CONCLUSION: Most patients with HBeAg seroconversion became inactive carriers with very good prognosis. The risk of liver-related mortality in Caucasian adults with CH is strongly related with sustained disease activity and ongoing high level of HBV replication independently of HBeAg status.

Evidence of Neospora caninum DNA in wild rodents.

Seventy-five house mice (Mus musculus), 103 rats (Rattus norvegicus) and 55 field mice (Apodemus sylvaticus) from North-West Italy were PCR analysed for Neospora caninum infection. Brain, kidney and muscle tissues collected from the above mentioned animals were tested by PCR using Np6 and Np21 primers. The brain tissue from 2 house mice and 2 rats, the kidney from 4 rats, 1 house mouse and 1 field mouse and muscle from 10 rats, 8 house mice and 1 field mouse were tested positive for N. caninum. Sequencing showed a 96-97% identity of PCR products with N. caninum NC1 sequence. Our findings support previous report on house mouse and rat, and for the first time, provides the evidence of the infection also in field mice. Based on our data, it could be hypothesized that mice can act as a reservoir of N. caninum, and they can play a role in maintaining/spreading N. caninum infection also in the sylvatic cycle. The possibility that dogs could be infected by eating infected house mice suggests new opportunities for N. caninum prophylaxis and control.

Antibodies to Neospora caninum in stray cats from north Italy.

Antibodies to Neospora caninum were determined in serum samples from 282 stray cats coming from four colonies near Turin (north-west Italy). Sera were tested using a Neospora Agglutination Test (NAT). Seroprevalence was 24.8% at 1:80, 12.8% at 1:160 and 5.3% at 1:320 dilution. Seroprevalence in females and males, in different colonies and in different age classes, did not differ significantly. Our results confirm that domestic cats are exposed to N. caninum and the observed seroprevalences suggest that risk of exposure is high and N. caninum should be considered in differential diagnosis in cats with neurological signs.

Congenital hypoplastic anaemia in a patient with a new multiple congenital anomalies-mental retardation syndrome.

We report on a girl with congenital hypoplastic anaemia, "coarse" face, generalized hypertrichosis with scalp hypotrichosis, short fifth finger, hypoplastic toenails, and mental retardation. A sister of the proposita, who died at the age of 1 year, had severe congenital anaemia, hypoplastic fingernails, low birth weight, failure to thrive, and repeated upper respiratory tract infections. Based on family history, we suspect that hypoplastic anaemia and the same multiple congenital anomalies-mental retardation syndrome (MCA/MR) were also present in this sister. To the best of our knowledge, this patient represents the first report of congenital hypoplastic anaemia and such a complex MCA/MR syndrome, probably inherited as an autosomal recessive trait.

Secondary acute promyelocytic leukemia with t(8;21) and t(9;22) at onset and loss of the Philadelphia chromosome at relapse.

The translocation (8;21) is a typical marker of the M2 subtype of acute nonlymphocytic leukemia, whereas the Philadelphia (Ph) chromosome is predominantly associated with chronic myelogenous leukemia, and seldom with immature acute leukemias, either lymphoblastic or nonlymphoblastic. Furthermore, the association between t(8;21) and a Ph in the same cell is extremely rare. We present a case of secondary acute promyelocytic leukemia with a karyotype 46,XY,t(8;21), t(9;22) at onset. At relapse the patient lost the Ph, while maintaining the t(8;21). This apparently paradoxical pattern of cytogenetic features and evolution is discussed.

A short-term in vitro drug sensitivity assay in pediatric malignancies.

A short-term in vitro chemosensitivity test has been applied to 131 specimens from 109 children with various malignancies. The clinical outcome was correlated with in vitro percentage inhibition (PI) of DNA synthesis by the drug tested. In 62 correlations, PIs of responsive and of resistant patients differed significantly, with a cut-off value of 37%. The test predicted clinical resistance in 53/54 cases, with a specificity of 98%, and clinical response in 8/8 cases, with a sensitivity of 100%. This test can be useful particularly in predicting chemoresistance in relapsing patients, to avoid unnecessary toxicity.

Urinary nitrite/nitrate concentrations and total antioxidant capacity in patients with chronic hepatitis C in therapy with interferon and ribavirin.

In this study we tried to elucidate the role of nitric oxide (NO) in chronic hepatitis C in relation to antioxidant status, since the mechanisms by which hepatitis C virus (HCV) causes cell damage and the factors underlying its resistance to therapy are not well understood. Before and after one and six months of therapy with alpha-interferon and ribavirin, we measured nitrite/nitrate urinary levels, total antioxidant capacity and selenium serum concentrations in 14 patients with chronic hepatitis C and in 9 healthy subjects. Before therapy, mean urinary nitrite/nitrate levels of patients were not different from those of healthy subjects, but after a 6-month treatment with alpha-interferon and ribavirin, these NO metabolites were higher in virologically negative patients (responders). Moreover, while no changes in selenium were observed in all patients, total antioxidant capacity was significantly higher in non-responders and well correlated with hyperuricemia (due to cell damage) observed in these subjects. Instead, uric acid decreased as free molecule in serum in responders, while we found the excretion of high NO levels as nitrite/nitrate. Our data allow us to hypothesize a role for NO as predictive of the success of therapy, since nitrite/nitrate increase in the urine of some patients precedes disappearance of the virus observed at the end of therapy.

IL28B polymorphisms, IP-10 and viral load predict virological response to therapy in chronic hepatitis C.

BACKGROUND: Hepatitis C virus (HCV) is a major cause of chronic liver disease, cirrhosis and hepatocellular carcinoma and the identification of the predictors of response to antiviral therapy is an important clinical issue. AIM: To determine the independent contribution of factors including IL28B polymorphisms, IFN-gamma inducible protein-10 (IP-10) levels and the homeostasis model assessment of insulin resistance (HOMA-IR) score in predicting response to therapy in chronic hepatitis C (CHC). METHODS: Multivariate analysis of factors predicting rapid (RVR) and sustained (SVR) virological response in 280 consecutive, treatment-naive CHC patients treated with peginterferon alpha and ribavirin in a prospective multicentre study. RESULTS: Independent predictors of RVR were HCV RNA <400 000 IU/mL (OR 11.37; 95% CI 3.03-42.6), rs12980275 AA (OR 7.09; 1.97-25.56) and IP-10 (OR 0.04; 0.003-0.56) in HCV genotype 1 patients and lower baseline γ-glutamyl-transferase levels (OR=0.02; 0.0009-0.31) in HCV genotype 3 patients. Independent predictors of SVR were rs12980275 AA (OR 9.68; 3.44-27.18), age <40 years (OR=4.79; 1.50-15.34) and HCV RNA <400 000 IU/mL (OR 2.74; 1.03-7.27) in HCV genotype 1 patients and rs12980275 AA (OR=6.26; 1.98-19.74) and age <40 years (OR 5.37; 1.54-18.75) in the 88 HCV genotype 1 patients without a RVR. RVR was by itself predictive of SVR in HCV genotype 1 patients (OR 33.0; 4.06-268.32) and the only independent predictor of SVR in HCV genotype 2 (OR 9.0, 1.72-46.99) or genotype 3 patients (OR 7.8, 1.43-42.67). CONCLUSIONS: In HCV genotype 1 patients, IL28B polymorphisms, HCV RNA load and IP-10 independently predict RVR. The combination of IL28B polymorphisms, HCV RNA level and age may yield more accurate pre-treatment prediction of SVR. HOMA-IR score is not associated with viral response.

Seroprevalence of antibodies to Neospora caninum in urban and rural dogs in north-west Italy.

Sera were collected from 490 dogs from north-west Italy. One hundred and eighty-eight dogs were urban, while 302 dogs were rural. Among the latter, 190 were shepherd dogs and 112 were cattle farm dogs. Sera were tested for the presence of antibodies against Neospora caninum using the Neospora agglutination test. Seroprevalence at 1/40, 1/80, 1/160 dilution titres was significantly higher in rural (36.4%, 19.5%, 9.9% respectively) than in urban dogs (20.2%, 10.6%, 4.8% respectively). Seroprevalence did not differ significantly in males and females. In shepherd dogs, prevalence increased according to dogs' age, thus suggesting a post-natal exposure by horizontal transmission. The observed higher seroprevalence in rural dogs suggests the importance of lifestyle and alimentary habits (i.e. aborted foetuses, placentas and small mammals) in the acquisition of N. caninum infection. Our results confirm that dogs are exposed to N. caninum and play an important role in the epidemiology of N. caninum.

Insight into molecular changes of the FIX protein in a series of Italian patients with haemophilia B.

Deficiency or dysfunction of factor IX FIX leads to haemophilia B (HB), an X-linked, recessive, bleeding disorder. On a molecular basis, HB is due to a heterogeneous spectrum of mutations spread throughout the F9 gene. In several instances, a cause-effect relation has been elucidated, in others predicted possibilities have been offered by crystallography inspection and by software-constructed models of the protein. The aim of this study was to contribute to the understanding of HB molecular pathology. The F9 missense mutations we identified in 21 unrelated Italian HB patients by direct sequencing of the whole F9 coding regions were inspected for the causative effect they provoked on the ensuing transcript, and on the protein structure. Each alteration was studied in order to: (i) characterize the defect on the basis of the nature of the mutation; (ii) identify the predicted defect that is induced in the gene and (iii) speculate about the potential, detrimental effects which upset the protein functionality through an idealized FIX model. The resulting data may further contribute to the comprehension of the mechanisms underlying the disease.

Active and passive rubidium influx in normal human lenses and in senile cataracts.

The aim of this study was to investigate the mechanisms responsible for the osmotic imbalance which occurs in a majority of human senile cataracts. Active and passive influx of rubidium has been determined in vitro in human lenses both normal and cataractous. It is concluded that the active transport of cations is on the average normal in senile cataractous lenses. It is possible that the activity of the cation pump is defective in a few cataractous lenses but no direct evidence of this can be given. The results indicate that cryo-extracted lenses may be utilized in this type of study provided that cryo-treatment during surgery is kept at the lowest possible level.

Detection of megakaryocyte colonies in plasma clot cultures by immunoenzymatic staining.

In vitro induced megakaryocytic differentiation/maturation of megakayocyte (meg) progenitors represents an important tool for investigating cytokine-induced in vitro thrombocytopoiesis. We have developed an assay which allows the in situ study of human meg progenitor-derived colonies, cultured on a plasma clot in the presence of cytokines. Plates were immunostained by using an anti-alpha IIb beta 3 monoclonal antibody and an alkaline phosphatase-labeled secondary antibody. alpha IIb beta 3-bearing cells were stained an intense red and were clearly differentiated from the negative cells. Processed plates were stable for some weeks at 4 degrees C. The described procedure is easy to perform and allowed us to enumerate the meg colonies and assess colony morphology and cell ploidy.

Bone-marrow T-lymphoid cells: a simple method for enriched specimen.

A simple method for immunological characterization of lymphoblasts from hypoplastic or fibrotic bone marrow aspirates is described. The method is quite suitable for immunological diagnostic purposes in classifying acute lymphoblastic leukemias (ALL) at onset of the disease. In fact, T-, common and classified ALL appear to have a different prognosis with consequent different patient management.

Methisoprinol effect on enriched B and T lymphocyte populations stimulated with phytohemagglutinin.

The Authors investigated the effect of methisoprinol on separated B and T lymphocytes from 8 healthy donors. B or T treated cells, recombined with T or B untreated cells, were subsequently mitogen stimulated. Proliferative response resulted significantly increased, in comparison to a control, when T lymphocytes were preincubated with the drug, while treating only B lymphocytes led to a decrement. These data confirm the hypothesis of a direct action of the drug on T lymphocytes, but are also consistent with its influence on B cells too, even though with an opposite effect.

Methisoprinol effect on enriched B and T lymphocyte populations stimulated with phytohemagglutinin.

The Authors investigated the effect of methisoprinol (Inosiplex) on separated B and T lymphocytes from 8 healthy donors. B or T treated cells, recombined with T or B untreated cells, were subsequently mitogen stimulated. Proliferative response resulted significantly increased, in comparison to a control, when T lymphocytes were preincubated with the drug, while treating only B lymphocytes led to a decrement. These data confirm the hypothesis of a direct action of the drug on T lymphocytes, but are also consistent with its influence on B cells too, even though with an opposite effect.

Acute lymphoblastic leukemia in Noonan syndrome: report of two cases.

Only two cases of Noonan Syndrome (NS) associated with tumor have previously been reported. The authors describe two new cases of NS with acute lymphoblastic leukemia (ALL), which were part of a series of 370 consecutive ALL untreated patients.

Properties of lymphocytes in chronic HBAg-positive hepatitis. An approach to pathogenetic problems.

Lymphocytes from 22 children affected by chronic hepatitis were studied and a comparison was made with a control group of 32 healthy children in the same age range with respect to PHA dose-response curves. E-rosette formation and membrane immunoglobulin (Ig). From the results it appeared that PHA dose-response curves were displaced to the left (low PHA dose), DNA synthesis was significantly higher for all PHA dosages tested, and that the plasma of chronic hepatitis patients acted neither as inhibitor nor stimulator. The percentage of E-rosette-forming cells decreased, but the total number was normal. The mean percentage of Ig-bearing cells was normal but the single values were spaced over a wide range. It can be assumed that, at least in the chronic stage, there is an enhanced response to PHA and an increased percentage of cells without markers either for T or for B cells.

Aspecific transfer factor in children with Hodgkin's disease.

Six children suffering from Hodgkin's disease (HD), in different stages and free from chemo- and radiotherapy from at least four weeks, were treated with transfer factor (TF). PPD skin test, PHA-responsiveness, E-rosettes, B-lymphocytes were checked before TF therapy, after 6 and 9 TF doses and compared to the data at the onset of the disease. Two children with HD who did not receive TF, were examined as controls. PPD skin tests, PHA-responsiveness, E-rosettes did not ameliorate following TF treatment, while it has been noticed an increase in B-lymphocytes, both in percentage and absolute number. The Authors conclude that TF might have induced a slight B lymphoproliferation.