Costanza Mazzeo

Ten year incidence of HCV infection in northern Italy and frequency of spontaneous viral clearance

Background: Little is known of the incidence of hepatitis C virus (HCV) infection, and the frequency of spontaneous viral clearance in the general population is unknown. We conducted an epidemiological study in two Apennine towns in northern Italy. Methods: Anti-HCV (ELISA and RIBA third generation) and HCV-RNA by polymerase chain reaction were tested in thawed sera from an adult general population of Loiano-Monghidoro in 1986 and 1996, obtained in the context of the MICOL (Multicenter Italian Study on Cholelithiasis). In 1999, anti-HCV positive subjects and sex and age matched controls were recalled in order to identify risk factors for acquiring HCV infection and to assess the family composition of anti-HCV+ subjects. Results: For 1646 subjects, sera were available from both 1986 and 1996 (mean age in 1986 43 (0.39) years). In 1986, 57 (3.46%) subjects were HCV antibody positive (HCV-Ab+). Eight new cases were recorded in 1996: adult incidence was 50.3 cases/100 000 inhabitants/year. Fifty three of 63 (84.1%) HCV-Ab+ sera were also HCV-RNA+. Genotype 2a/2c accounted for 44% and 1b for 47.0% of cases. HCV-Ab+ subjects had higher serum levels of alanine aminotransferase with respect to controls (p<0.005), as did subjects infected with genotype 1 with respect to those with genotype 2 (p<0.05). Eleven of 65 (16.9%) HCV-Ab+ subjects spontaneously cleared HCV-Ab; 7/11 also lost HCV-RNA− in both serum and leucocytes. Sixteen anti-HCV+ subjects belonged to families containing more than one infected member. Married couples accounted for 10 of these 16 subjects. In four of these five married couples, HCV genotype was identical in the two spouses. Conclusions: In rural northern Italy, the adult incidence of HCV is approximately 50 cases/100 000 inhabitants/year. Our findings suggest that as many as 17% of infected subjects may spontaneously clear HCV-Ab. Interfamilial transmission seems to have a role in the spread of infection.

Sluggish small bowel motility is involved in determining increased biliary deoxycholic acid in cholesterol gallstone patients.

OBJECTIVE:Our aim was to establish whether small intestine transit time is defective in subjects with cholesterol gallstones.METHODS:We enrolled 10 patients (eight women, two men; mean age, 48.7 yr; mean body mass index [BMI], 22.4 Kg/m2) with recently diagnosed cholelithiasis, with no liver pathology, who were not taking any drugs, and 11 comparable healthy volunteers (eight women, three men; mean age, 46.2 yr; mean BMI, 22.7 Kg/m2), who served as controls. All subjects underwent orocecal (by starch breath test technique and serum assays of salazopyrin), oroileal (by serum assays of tauroursodeoxycholic acid), and duodenoileal (by serum assays of taurocholic acid) transit times; cholesterol saturation index; and bile acid composition and gallbladder motility studies (by ultrasound). For serum assays, blood samples were collected over a period of 7 h. Gallbladder motility and orocecal transit time were evaluated simultaneously.RESULTS:All four means of assessing transit time gave longer times in cholesterol gallstone patients than in controls: orocecal transit time (salazopyrin) = 366 ± 13 vs 258 ± 16 min, p < 0.0005; orocecal transit time (starch breath test) = 415 ± 139 vs 290 ± 15 min, p < 0.01; duodenoileal transit time: 272 ± 23 vs 205 ± 23 min, p < 0.03; and oroileal transit time: 308 ± 18 vs 230 ± 19 min, p < 0.009. Cholesterol gallstone patients showed an increase in percent molar biliary deoxycholic acid (30%± 4.5%vs 16%± 1.3%, p < 0.02) and a decrease in percent molar cholic acid 32%± 2.2%vs 40%± 1.3%, p < 0.03) and chenodeoxycholic acid (34%± 3%vs 41%± 1.8%, p < 0.03), compared with controls; patients also had greater percent molar biliary cholesterol. A linear relationship (r2= 0.6324, p= 0.0012) between biliary deoxycholic acid and small bowel transit time was found. Residual gallbladder volumes were larger in cholesterol gallstone patients (11.38 ± 1.27 vs 7.55 ± 0.39 ml, p < 0.04), whereas basal gallbladder volumes, although higher, did not reach statistical significance (24.25 ± 2.41 vs 19.98 ± 1.63 ml; p= ns).CONCLUSIONS:This study confirms that patients with cholesterol gallstones have delayed small bowel transit, defective gallbladder motor function, and increased biliary deoxycholic acid. Delayed small bowel transit may contribute to supersaturation of bile with cholesterol by increasing deoxycholic acid production.

Ten-year incidence of HCV infection in northern Italy and frequency of spontaneous viral clearance

BACKGROUND Little is known of the incidence of hepatitis C virus (HCV) infection, and the frequency of spontaneous viral clearance in the general population is unknown. We conducted an epidemiological study in two Apennine towns in northern Italy. METHODS Anti-HCV (ELISA and RIBA third generation) and HCV-RNA by polymerase chain reaction were tested in thawed sera from an adult general population of Loiano-Monghidoro in 1986 and 1996, obtained in the context of the MICOL (Multicenter Italian Study on Cholelithiasis). In 1999, anti-HCV positive subjects and sex and age matched controls were recalled in order to identify risk factors for acquiring HCV infection and to assess the family composition of anti-HCV+ subjects. RESULTS For 1646 subjects, sera were available from both 1986 and 1996 (mean age in 1986 43 (0.39) years). In 1986, 57 (3.46%) subjects were HCV antibody positive (HCV-Ab+). Eight new cases were recorded in 1996: adult incidence was 50.3 cases/100 000 inhabitants/year. Fifty three of 63 (84.1%) HCV-Ab+ sera were also HCV-RNA+. Genotype 2a/2c accounted for 44% and 1b for 47.0% of cases. HCV-Ab+ subjects had higher serum levels of alanine aminotransferase with respect to controls (p<0.005), as did subjects infected with genotype 1 with respect to those with genotype 2 (p<0.05). Eleven of 65 (16.9%) HCV-Ab+ subjects spontaneously cleared HCV-Ab; 7/11 also lost HCV-RNA- in both serum and leucocytes. Sixteen anti-HCV+ subjects belonged to families containing more than one infected member. Married couples accounted for 10 of these 16 subjects. In four of these five married couples, HCV genotype was identical in the two spouses. CONCLUSIONS In rural northern Italy, the adult incidence of HCV is approximately 50 cases/100 000 inhabitants/year. Our findings suggest that as many as 17% of infected subjects may spontaneously clear HCV-Ab. Interfamilial transmission seems to have a role in the spread of infection.

Serological response to pgp3 protein in animal and human chlamydial infections.

Specific antibodies to plasmid-encoded protein pgp3 are known to be encountered in human Chlamydia (C.) trachomatis infections. In order to verify whether antibodies to this protein could be developed in animals infected with plasmid-carrying chlamydial strains, 454 animal sera were examined using a home-made pgp3 protein ELISA and Western blots (WB) of recombinant pgp3 protein from Chlamydophila (Cp.) psittaci. Likewise, 50 human sera were tested by ELISA and WB of recombinant pgp3 from C. trachomatis. The reactivity against pgp3 protein was compared to the reactivity against chlamydial elementary bodies (EBs) detected by microimmunofluorescence (MIF) test. The presence of pgp3-specific antibodies was demonstrated in most ducks and pigeons with Cp. psittaci infection detected by MIF, as well as in the majority of symptomatic cats and pigs infected with Cp. felis and C. suis, respectively, which reacted at high titres to Cp. felis and C. suis EBs by MIF. Moreover, most of the sera collected from patients with C. trachomatis culture-confirmed infection and seropositive to C. trachomatis by MIF, presented antibodies specific to C. trachomatis pgp3 recombinant protein. Therefore, pgp3 protein could be a useful marker of chlamydial infections in animals, as well as in humans.

Efficacy of ursodeoxycholic acid in association with α-interferon for chronic hepatitis C in α-interferon non-responder patients

BACKGROUND The aim of the present study was to evaluate the effect of combined treatment with alpha-interferon (alpha-IFN) and ursodeoxycholic acid (UDCA) on liver function tests and serum HCV-RNA in patients with chronic hepatitis C who had not responded to alpha-IFN alone. METHOD One hundred and three patients (60 men, 43 women, mean age 49 +/- 1.3 years) who had not responded (both HCV-RNA positive and increased serum ALT levels) to 4 consecutive months of treatment with alpha-IFN (3 MU three times weekly) were randomly assigned to receive UDCA (IFN-UDCA, 53 patients, 600 mg/day) in addition to the same alpha-IFN dose, or to continue alpha-IFN alone (IFN-controls, 50 patients). After stopping alpha-IFN, patients who had received UDCA continued to receive UDCA for an additional 6-month period. The two groups were comparable for sex, basal ALT, basal yGT, genotype distribution and liver histology, while mean age was lower in controls (53 +/- 1.8 vs 46 +/- 1.8 years; P< 0.01). RESULTS Twenty (38%) out of 53 IFN-UDCA patients had normal ALT, compared with only six (12%) out of 50 IFN-control patients (P < 0.01). HCV-RNA became undetectable in four IFN-UDCA patients. Three months after withdrawal of alpha IFN, 15 IFN-UDCA responders, but none of the IFN-controls, had normal ALT values (P< 0.01); 6 months after withdrawal, nine IFN-UDCA responders still had normal ALT (P= NS) and HCV-RNA was still undetectable in four of them. Portal and periportal inflammation showed a statistically significant improvement (Fishers exact test P< 0.01) in IFN-UDCA patients as compared with IFN-controls, while no effect was observed on portal fibrosis. CONCLUSIONS These data demonstrate that UDCA improves the response rate to alpha-IFN. Furthermore, in 8% of IFN-UDCA patients the response rate was sustained and associated with HCV-RNA clearance.

Methylprednisolone Administration in Primary Biliary Cirrhosis Increases Cholic Acid Turnover, Synthesis, and Deoxycholate Concentration in Bile

As immunosuppressive agents, corticosteroids maybe considered an appropriate treatment for primarybiliary cirrhosis, even if bone loss and other sideeffects may occur. We studied biliary lipid metabolism in 10 nonicteric patients, with histologicallyproven primary biliary cirrhosis (stage I-IV). Weadministered methylprednisolone (24 mg daily) for 30days to ascertain its effects on biliary lipidmetabolism, which are largely still unknown. All patientsunderwent a 30-day drug-washout period before enteringthe trial. The following parameters were studied beforeand after methylprednisolone treatment: serum biochemistry; cholic acid pool size, kineticsand synthesis; biliary lipid secretion; biliary bileacid pattern; biliary lipid molar percentage; andcholesterol saturation index. Methylprednisolone induced a statistically significant (Wilcoxon ranktest) increase in cholic acid turnover (from 0.26± 0.04 to 0.50 ± 0.05 K/day, P = 0.005)and synthesis (from 0.42 ± 0.12 to 0.78 ±0.11 mmol/day, P = 0.04), and in bile deoxycholic acid molarpercentage (from 19.4 ± 2.7 to 30.6 ± 4.4%molar, P = 0.01). On the other hand, a significantdecrease in biliary cholesterol molar percentage (from7.9 ± 0.7 to 6.4 ± 0.5 % molar, P =0.005), cholesterol saturation index (from 1.11 ±0.11 to 0.95 ± 0.07, P = 0.05), and biliarycholesterol secretion (from 64.7 ± 5.4 to 53.0± 4.5 μmol/hr, P = 0.005) was observed. These findings show thatshort-term administration of methylprednisolone inpatients with primary biliary cirrhosis does not induceexpansion of the cholic acid pool but increases cholicacid synthesis and turnover, as well as intestinalproduction of deoxycholic acid. If long-term treatmentis considered, the beneficial immunosuppressive effectsof corticosteroids have to be weighed against the hepatotoxic properties of deoxycholicacid.

Feline chlamydiosis: a seroepidemiological investigation of human beings with and without contact with cats

THE transmission of chlamydial infection from cats to human beings was first demonstrated by Schachter and others (1969), who isolated chlamydia from a cat with signs of conjunctivitis and rhinitis, and from its owner who had developed acute follicular conjunctivitis. The isolate taken from the cat’s owner was then used to reproduce conjunctivitis in a healthy cat. There have been very few reports on the same topic since (Shewen and others 1978, Hartley and others 2001), possibly because the transmission of chlamydial conjunctivitis from cats to human beings is a rare occurrence, or more likely because such cases are underestimated as a result of failed isolation or inadequate investigation (Longbottom and Coulter 2003). Close contact is needed for transmission of the infection to take place, so cat owners and professionals who work with cats (for example, veterin arians, employees of catteries and cat breeders) are most at risk. A seroepidemiological survey carried out in Japan to evaluate the prevalence of feline chlamydiosis in different human populations found a significantly higher incidence of seropositivity for Chlamydophila felis in veterinarians working with small animals (8·8 per cent) than in the general population (1·7 per cent) (Yan and others 2000). Other surveys have reported that cat owners whose pets had clinical signs of chlamydiosis did not have ocular signs of chlamydial infection (Studdert and others 1981, Wills 1986). This short communication assesses the circulation of C felis in human beings with and without contact with cats. A total of 303 sera samples from human beings were tested: 164 from individuals who had had contact with a cat (85 veterinarians and 79 owners), and 139 from subjects who had had no such contact. The sera were tested for the presence of anti-C felis antibodies by an indirect microimmunofluorescence test (MIF). The test was performed, according to the method of Wang and Grayston (1986), using the purified elementary bodies (EBs) of the Italian strain of C felis (FEIS-M) as antigen, isolated from a conjunctival swab from an infected cat. The EBs were purified by sucrose density-gradient ultracentrifugation by the method of Fukushi and Hirai (1988). The presence of anti-C felis antibodies was assessed using a fluorescein-conjugated goat anti-cat immuno globulin G serum (Euroclone). Sera were screened at 1:32 dilution. Serial two-fold dilutions of the sera that were positive at 1:32 dilution were tested to determine the antibody titre. The reciprocal of the highest serum dilution to show an apple-green fluorescence of EBs was considered as the antibody titre. To verify cross-reactions with other chlamydia species, the MIF was performed on all sera showing an antibody titre for C felis of 32 or higher using, as antigens, the purified EBs of the avian reference strain 6BC of Chlamydophila psittaci and the Italian strain FB of Chlamydophila pneumoniae isolated from a patient with pneumonia. The same sera were tested against purified EBs of C felis, C psittaci and C pneumoniae by immunoblotting analysis (Fukushi and Hirai 1988), to evaluate their reactivity to proteins of the three chlamydial species. Antibodies to C felis were detected in 23 (7·6 per cent) of the 303 samples tested, with antibody titres ranging from 32 to 128. Of these positive samples, 13 were from human beings that had had contact with cats (seven veterinarians and six owners), while 10 sera were from individuals that had had no contact with cats. All the C felis-positive sera were also positive for C pneumoniae (titres between 32 and 512); two sera (from veterinarians) were positive for C psittaci at a low titre (1:32). By immunoblotting, all 23 sera were found to have a strong reactivity with C pneumoniae antigens, including the species-specific 98 kDa protein, but had a weak reactivity to C felis and C psittaci proteins (Fig 1). The cross-reactivity demonstrated between C felis and C pneumoniae showed there was no circulation of C felis in the different groups, which is not surprising despite the known presence of C felis in stray and household cats in Italy (Di Francesco and others 2004). The transmission between cats and human beings is possible, but only in the case of a massive contamination of the conjunctiva (for example, rubbing of the eyes with hands contaminated from the handling of symptomatic cats). Therefore, the adoption of common hygiene practices should prevent such transmission taking place (Blanchart 1994). The survey also showed a cross-reactivity between C felis and C pneumoniae, but no specific reactivity to C felis EBs in the sera examined. This suggests that, in the course of seroepidemiological surveys on feline chlamydiosis in the human population, cross-reactions between C felis and C pneumoniae should always be evaluated because of the higher seroprevalence of C pneumoniae in the adult human population (Grayston and others 1990). Isolation of the aetiological agent remains the gold standard for the diagnosis of chlamydial infection.