White mulberry supplementation as adjuvant treatment of obesity.

Body weight is controlled by our genes and managed by a neuro-hormonal system, in particular by insulin and glucagon. The meristematic extract of Japanese white mulberry blocks the alpha-glucosidase and then the intestinal hydrolysis of polysaccharides, thereby reducing the glycaemic index of carbohydrates. The target of our research was to evaluate the adjuvant slimming effect of the extract of white Japanese mulberry in the dietetic treatment of some patients who are obese or overweight. 46 overweight people were enrolled and divided into two subgroups: the subjects of both subgroups were given an identical balanced diet of 1300 kcal: subjects of the subgroup alpha received 2400 mg of white Japanese mulberry extract, the subgroup b subjects receive placebo. Each subgroup was followed-up every 30 days at 30, 60 and 90 days of treatment. Both in the periodic inspections and in the final inspection measurements of body weight and waist circumference in all the subjects and thigh circumference in women only were repeated. All subjects repeated blood tests. In the subgroup alpha, weight loss was about 9 kg in 3 months, equal to approximately 10 percent of the initial weight, significantly higher than subgroup beta (P<0.0001); moreover, the plasma insulin and glucose curves of the volunteers in this subgroup at the end of the trial were lower than those performed at the time of enrolment. In the 20 women of the beta subgroup treated with only low-calorie diet and with placebo, weight reduction was globally of 3.2 kg, approximately equal to 3 percent of the initial weight; moreover, the blood glucose curves and the insulin curves showed a slight decline compared to baseline, but not so significantly as was the case for group alpha. Waist circumference and thigh circumference (in women) decreased in all participants, obviously more evidently in subjects who lost more kg. The extract of white Japanese mulberry may represent a reliable adjuvant therapy in the dietetic treatment of some patients who are obese or overweight.

Rationale for the infant and adolescent vaccination programmes in Italy.

In Italy in the 1980s, the incidence of acute hepatitis B was about 13 per 100,000, corresponding on average to 7500 new symptomatic cases per year was about 3%, making Italy an area of intermediate endemicity. HBV infection was also associated with 12 per 100,000 deaths from cirrhosis and with 5.1 per 100,000 deaths from hepatocellular carcinoma. In view of the large numbers of pregnant women who were hepatitis B surface antigen (HBsAg)-positive, selective hepatitis B vaccination of all newborns to these mothers and of other high-risk groups was introduced in 1983. Compliance was high among the newborns but low in other high-risk groups. Hepatitis vaccination was adopted in Italy in 1991, including each year all newborns, all adolescents aged 12 years and other high-risk groups. Compliance has been nearly 95% for newborns and 80% for adolescents. Since the introduction of vaccination, both the incidence of acute hepatitis B and the prevalence of HBV carriage have fallen, the latter from 3.4% in 1985 to 0.9% in 1996. There is good evidence that these decreases are mainly the result of the vaccination programmes. Although the full economic impact cannot yet be assessed, about 18,000 cases of acute HBV infection have been prevented over the 6 years since starting the mass vaccination programme, with cost savings of about US$ 244,308,000.

From hepatitis B to hepatitis A and B prevention: the Puglia (Italy) experience.

The incidence of hepatitis B virus infection in Italy is 10 per 100, 000 population, with most cases occurring in young adults. Vaccination against hepatitis B has been compulsory since 1991 for all newborns and 12-year-olds. In the Puglia region, this programme has reduced the incidence of hepatitis B from 7.4 per 100,000 population in 1990 to 2.4 per 100,000 population in 1996. The number of notified cases of hepatitis B in Puglia decreased from 212 in 1992 to 73 in 1997. As 50% of these cases occurred in young adults, the main aim of the current vaccination programme is to achieve high coverage rates among teenagers and young adults within the next few years. Although the incidence of hepatitis A is only about 5 per 100, 000 overall in Italy, Puglia is an area of intermediate endemicity with a seroprevalence of antibodies to hepatitis A virus (anti-HAV) of about 40% in 18-year-olds. The incidence of hepatitis A is up to 30 per 100,000 between the periodic outbreaks that occur every 2-4 years. Most notified cases occur in adolescents and young adults. The last outbreak of about 11,000 cases of hepatitis A in the Puglia region occurred in 1996-1997, mainly in the summer months in towns with harbours or near the coast. The most important risk factor was initially consumption of raw seafood, but later was personal contact, probably between children. A vaccination programme against hepatitis A was initiated in Puglia in 1997, aiming to vaccinate all infants of 15-18 months and all 12-year-olds against hepatitis A. Infants receive monovalent hepatitis A vaccine with the first dose of mumps/measles/rubella vaccine. Monovalent hepatitis vaccine can be given with the second and third doses of hepatitis B vaccine in 12-year-olds, but use of combined hepatitis A and B vaccine is recommended to aid compliance and reduce the commitment of physician/nurse time. Vaccination can be performed in school.

Immunization programme against hepatitis B virus infection in Italy: cost-effectiveness.

In Italy in the 1980s the extent of viral hepatitis B infection was on average about 11,000 symptomatic cases of acute viral hepatitis (AVH) per year (19/100,000 inhabitants). The prevalence of HBsAg carriers in the general population was about 3% and in pregnant women 2.4%. 64,000 people were affected by chronic viral hepatitis (CVH) or cirrhosis (prevalence rate 112/100,000) and 3400 by hepatocellular carcinoma (HCC) (prevalence rate 5.9/100,000). To reduce these HBV related pathologies in the Italian population, universal vaccination of newborn babies, 12-year old adolescents and high risk groups was implemented in 1991. The annual cost of this immunization is about 57 million 544 thousand USD: direct costs: 41 million 34 thousand USD; indirect costs: 16 million 510 thousand USD. Concerning the vaccination impact on HBV endemicity in Italy, we found a significant reduction of acute viral hepatitis incidence (4.2/100,000 in 1996 versus 19/100,000 in the '80s) and HBsAg carrier prevalence (0.9% in 1997 versus 3% in the '80s). As for the assistance and social cost of acute viral hepatitis occurring from 1991 to 1996 (17,608 cases) it was 238 million 908 thousand USD, while the cost for the same pathology in the years from 1985 to 1990 (35,614 cases) was 483 million 216 thousand USD. Thus, the saving during the years of the vaccination was evaluated in 244 million 308 thousand USD. At the moment, we have no information about the reduction in chronic sequelae of HBV pathology as an effect of the vaccination, because the incidence of this pathology generally starts to appear after 15 years (in our case in 2006).

Antibodies to the E2 protein of GB virus C/hepatitis G virus: low prevalence in Asian countries.

Epidemiological investigations of GB virus C (GBV-C)/hepatitis G virus (HGV), an infectious agent discovered in 1995/1996, are facilitated by a recently developed immunoassay for the detection of antibodies to the viral envelope 2 protein (anti-E2). We used this assay to establish GBV-C/HGV prevalence in seven European, African, and Asian countries. A total of 1579 serum samples from healthy adults lacking prior exposure to known parenteral risk factors was screened. Anti-E2 positivity ranged from 13.6% (Italy) to 7.7% (Mauritius) in the European and African countries investigated. Anti-E2 prevalence was exceedingly low in the Philippines and Sri Lanka. This observation might be attributable to socio-economical and demographic factors.

Distinct prevalence of antibodies to the E2 protein of GB virus C/hepatitis G virus in different parts of the world.

Since the identification of the new human virus, GB virus C (GBV-C)/hepatitis G-virus (HGV), in 1995/1996, reverse transcription polymerase chain reaction remained the sole available diagnostic tool for GBV-C/HGV infection. Recently, a serologic test based on the detection of antibodies to the putative envelope protein 2 (anti-E2) has been introduced. We used this assay for a seroepidemiological survey including 3,314 healthy individuals from different parts of the world, 123 patients from Germany who were suspected to have an increased risk of acquiring GBV-C/HGV infection, 128 multiple organ donors, and 90 GBV-C/HGV RNA positive persons. In European countries, anti-E2 seropositivity ranged from 10.9% (Germany) to 15.3% (Austria). In South Africa (20.3%) and Brazil (19.5%), even higher anti-E2 prevalence rates were recorded. In Asian countries like Bhutan (3.9%), Malaysia (6.3%), and the Philippines (2.7%), anti-E2 positivity was significantly lower. GBV-C/HGV anti-E2 prevalence in potential "risk groups," i.e., patients on hemodialysis and renal transplant recipients, did not vary significantly from anti-E2 seroprevalence in German blood donors. Anti-E2 and GBV-C/HGV RNA were found to be mutually exclusive, confirming the notion that anti-E2 has to be considered as a marker of past infection.

Long-term epidemiological survey of hepatitis B virus infection in a hyperendemic area (Afragola, southern Italy): results of a pilot vaccination project.

In 1983, a pilot project of universal hepatitis B vaccination was introduced in a hyperendemic area in southern Italy (Afragola) and is ongoing to date. In this area before the start of vaccination, we found significant evidence of HBV endemicity: the acute viral hepatitis B incidence in the general population averaged 63/100,000; the HBsAg and anti-HBc prevalence rates were 13.4% and 66.9%, respectively; there was involvement of hepatitis B virus (HBV) in 48.1% of chronic liver pathologies (46.3%) in chronic viral hepatitis, 49.5% in cirrhosis and 71.7% in hepatocellular carcinoma cases). We studied the acute viral hepatitis incidence during the vaccination period from 1983 to 1997 and compared the HBsAg and anti-HBc prevalences in 1978 to those in 1997, after 15 years of vaccination. The HBV-related chronic pathology prevalence was also studied. We found a remarkable drop in the acute viral hepatitis incidence, from an average annually of 63/100,000 in the five years before vaccination to 3/100,000 in the last five years of vaccination. In addition, the HBsAg carrier prevalence in the general population decreased from 13.4% in 1978 to 3.7% in 1997. The percentage dropped in children and adolescents from 6.8% to 0.7%, in young people from 10.2% to 1.1% and in adults from 15.8% to 4.0%. The anti-HBc carrier prevalence, found to be 66.9% in 1978, was 34.2% in 1997. Finally, we found a much less significant involvement of HBV in chronic liver pathologies; in fact, it was present in only 18.2% of cases in 1997 and in 48.2% in 1982. In the light of the data, we can assert that universal hepatitis B vaccination has had a substantial effect on HBV endemicity in the Afragola area. We believe that the reduction found in the incidence of acute viral hepatitis B and HBV-related chronic liver pathologies is connected to the decrease in HBV carriers in the area, which therefore reduces the risk of contagion for the unvaccinated.

Sequence variability in the putative coding region of TT virus: evidence for two rather than several major types.

Recently a new human virus, TT virus (TTV) was identified in the serum of a patient with post-transfusion hepatitis of unknown aetiology. Comparative sequence analysis of a 222 nt fragment of ORF 1 of TTV was performed to assess the genomic variability of this virus. Phylogenetic analysis of the nucleotide sequences of 76 TTV isolates collected in 17 countries segregated them into two major groups: TTV 1 and TTV 2. The TTV 1 group comprised two distinct subgroups, which corresponded to previously described TTV subtypes 1a and 1b. The TTV 2 group was separated into four main branches, two of which included sequences previously provisionally attributed as TTV types 2 and 3. Bootstrap resampling, however, did not support the reliability of this grouping, suggesting that the isolates in the TTV 2 group should be considered as subtypes of a single type rather than different TTV types.

Anti-HBs responses in children vaccinated with different schedules of either plasma-derived or HBV DNA recombinant vaccine.

This study evaluated the immunogenic and protective effects of plasma-derived and DNA recombinant anti-hepatitis B virus vaccines administered to infants at various ages and with different vaccination schedules: 3 monthly doses in the first 3 months of life, 3 doses (at 3, 5 and 11 months) or 2 doses (at 1 and 3 months) or 2 doses (at 3 and 5 months). Anti-HBs (hepatitis B surface) and anti-HBc (hepatitis B core) markers were investigated twice: one month and ten years after vaccination in 261 children immunized with plasma-derived vaccine, and one month and five years after vaccination in 449 children immunized with DNA recombinant vaccine. In all groups, the appearance of anti-HBs protective levels one month after vaccination and their persistence in the following years were found in a larger number of subjects when the vaccine doses had been administered after the third month of life rather than in the first three months. Moreover, our results show that the reappearance of surface antibodies a week after the booster, in vaccinated children who became anti-HBs- in the years following vaccination, occurred in a larger number of subjects when the primary vaccination with 3 doses had been performed in the first quarter or with 2 or 3 doses in the second quarter. In contrast, protective levels of anti-HBs were found in a small number of children belonging to the group vaccinated with 2 doses in the first three months, and among them the majority seroconverted only one month after the booster. Anti-HBc was found 10 years after vaccination in only one child immunized with 2 doses of plasma-derived vaccine, and 5 years after vaccination in two children immunized with 2 doses of DNA recombinant vaccine. All these children were found to lose anti-HBs, and none of them had signs of disease or became a carrier. Based on these results, the disappearance, in some children, of protective levels of anti-HBs in the years following vaccination does not mean the loss of anti-HBV protection. In fact, the trial showed that they reacted immediately to booster stimulation, demonstrating a solid immunologic memory. Therefore, there may be no reason for giving booster injections when the vaccination of infants is carried out correctly.

Viral hepatitis in the Bhutanese population: preliminary results of a seroepidemiological investigation.

A seroepidemiological study of the prevalence of markers related to the most common forms of viral hepatitis was carried out in Bhutan on 1,666 healthy people of both sexes, from the general population. A group of 440 pregnant women were screened separately. Our results suggest that in Bhutan, hepatitis A and hepatitis B are widespread, while there is a low prevalence of hepatitis C and E. Anti-HAV (anti-hepatitis A virus) was found in all 171 tested subjects over 12 years of age, and anti-HBc (anti-hepatitis B core antigen) in 63.1% of 1,666 tested people. On the other hand, anti-HEV proved positive in 2.0% of 257 tested subjects, and anti-HCV in only 1.3% of 611 tested subjects. Hepatitis B surface antigen (HBsAg) was found in 5.9% of the sample from the general population (5.2% in children, 5.6% in young people and 6.3% in adults) and in 5.4% of the pregnant women. Furthermore, 29.1% of HBsAg-positive pregnant women were HBeAg- and HBV DNA-positive, too. Comparing the pregnant women's prevalence data to those found in children, we suggest that the main route of HBV transmission in the Bhutanese population is vertical, from mother to child; this finding is important for the implementation of a correct anti-HBV vaccination strategy in Bhutan.

Persistence of anti-HBs in children vaccinated against viral hepatitis B in the first year of life: follow-up at 5 and 10 years.

The persistence of anti-HBs protective levels in groups of children who had been immunized against Hepatitis B 5 and 10 years earlier, in their first year of life, has been studied. The results were analyzed according to the type of vaccine (both plasma-derived and DNA recombinant) and the number of doses administered (three or four doses). In addition, the protective effect of the vaccine in vaccinated subjects was studied in relation to the anti-HBc presence. The serologic results suggest that, in cohorts of children vaccinated 5 years earlier, a higher prevalence of subjects with anti-HBs protective levels was found, when the DNA recombinant vaccines were administered (97.6% for MSD Recombivax and 97.1% for SKF Engerix B); a lower one when the plasma-derived vaccine was used (80.4% Pasteur Merieux, Hevac B). Moreover, in cohorts of children vaccinated with plasma derived vaccine (hevac B) 10 years earlier, a higher prevalence of subjects with anti-HBs protective levels was demonstrated when four doses were administered (76.9%); a lower one when three doses were inoculated (57.5%). The absence of core antibody (anti-HBc) in responders to the vaccination shows the protective efficacy of both the DNA recombinant and plasma derived vaccines. On the other hand the presence of anti-HBc in some anti-HBs negative non-responders subjects shows the susceptibility of these people to infection. In anti-HBs negative vaccinated subjects the appearance of levels of anti-HBs in 95.9% of subjects, 1 week after the administration of a booster dose, demonstrates the presence of a solid immunologic memory.

Hepatitis B vaccination: universal vaccination of newborn babies and children at 12 years of age versus high risk groups. A comparison in the field.

From 1983 to 1993 two anti-hepatitis B vaccinal strategies were adopted in two small towns of Southern Italy at high incidence for HBV infections: Afragola (prevalence of HBsAg carriers of 13.4%) and Frattamaggiore (prevalence of HBsAg carriers of 12.9%). In Afragola, the universal vaccination of infants in their first year of life and adolescents at 12 years of age was carried out, while in Frattamaggiore the selective vaccination of high risk groups of population was introduced. During this study, the changes in the HBV infection endemicity in both towns has been tested by monitoring the incidence of new cases of viral hepatitis B and by the prevalence study of HBsAg carriers prior to and 10 years after the beginning of the immunization programme (1978-1993). The results suggest that universal vaccination of infants in the first year of life and adolescents at 12 years of age has a greater efficiency on the improvement on the endemic status of the infection in the general population in comparison with selective vaccination, when the incidence of new cases of disease and the prevalence of number of HBsAg and anti-HBc carriers in the two populations are considered.

Fusion of EBV with the surface of receptor-negative human hepatoma cell line Li7A permits virus penetration and infection.

Our preliminary data suggest that Epstein-Barr virus (EBV) is able to bind to and fuse with the surface membranes of hepatoma cell line Li7A. In order to obtain further evidence, we utilized the relief of rhodamine fluorescence to monitor whether fusion would also take place when Li7A cells were exposed to experimental conditions such as neutral or low pH. It is well known that for some viruses, protonation in the endosomal compartment is needed to trigger the fusion. We show, furthermore, that the rate and extent of fusion are not affected by pretreatment of the cells with agents known to elevate the lysosomal and ensodomal pH, such as chloroquine or NH4Cl (lysosomotropic agent). By indirect immunofluorescence assay, in addition, we confirmed the binding of the EBV to the Li7A cell surface membrane. We attempted finally to correlate the above processes with successful infection of Li7A cells by EBV detected using the polymerase chain reaction technique. In spite of the apparent lack of viral receptor CD21, these nonlymphoid cells appeared susceptible to EBV penetration and infection through fusion with the plasma membrane at the surface of the cells.

Effective antibody response in newborn babies living in Maldives to simultaneous vaccination against hepatitis B, poliomyelitis, diphtheria and tetanus.

The antibody responses of Maldivian infants early in their life to simultaneous immunization against hepatitis B virus, poliomyelitis, diphtheria and tetanus were investigated. The vaccines were given at 6, 10 and 14 weeks of age. Among 243 newborn babies from HBsAg-negative mothers, 103 received three doses of oral poliomyelitis (OPV) and diphtheria and tetanus (DTV) vaccines; 105 were similarly immunized but received in addition the recombinant hepatitis B vaccine (HBV); 35 were immunized with the HBV recombinant vaccine alone. The antibody response to all of the vaccines was effective. No significant differences among the groups were observed. Hepatitis B vaccination of infants neither affected nor was affected by the contemporary administration of OPV and DTV vaccines.

Successful mass vaccination against hepatitis B virus in a hyperendemic area in Italy.

A pilot model of vaccination against hepatitis B virus suitable for mass vaccination was performed from 1983-1989 in the hyperendemic area of Afragola, a town of 60,000 inhabitants near Naples. In this area of high prevalence of HBsAg carriers, a consistent reduction in the incidence of hepatitis B virus infections as well as chronic related liver complications was observed.

A pilot model of vaccination against hepatitis B virus suitable for mass vaccination campaigns in hyperendemic areas.

A hepatitis B vaccination campaign was carried out in a town of 60,000 inhabitants, Afragola, Campania, Italy, a hyperendemic area for hepatitis B where HBsAg prevalence was 13.4% and anti-HBc prevalence was 64.7%. This experimental pilot project aimed to reduce the incidence of both acute and asymptomatic viral hepatitis B and of related chronic liver complications. From 1983-1989, 8,400 subjects were vaccinated: 6,900 children up to 10 years of age and 1,500 subjects from 11-60 years of age. High seroconversion rates were observed: 99.0% in all children under one year of age, 96.0% in the older children, and 86.7% in adults. The rate of infection in Afragola has diminished from 63/100,000 in 1983 to 10/100,000 in 1989. Carriers of HBsAg decreased in the general population (7.3% compared to 13.4%), especially in children up to 10 years of age (1.0% compared to 9.0%). In babies who received hepatitis B vaccine at the same time as compulsory vaccinations compliance was 98% while it was 80% in babies who were vaccinated separately. In June 1991 the Italian Parliament promulgated a decree which imposes hepatitis B vaccination for all newborn babies at 3, 5, and 11 months of age, at the same times as the mandatory childhood vaccinations (diphtheria, tetanus, and polio) according to a new protocol (Piazza scheme) which has been in use since January 1987 in our pilot vaccination campaign in Afragola.