Serological reactivity and viral genotypes in hepatitis C virus infection.

Patients infected with hepatitis C virus (HCV) were examined with four commercial HCV immunoblotting assays and for anti-GOR antibody to ascertain whether serological findings varied with the genotype of the infecting virus. The results indicate that patients infected with different HCV genotypes tend to show different immunoblotting profiles, mainly due to a low prevalence of antibodies to the viral region NS4 in patients infected with genotypes III and IV. Differences were more evident with second- than with third-generation assays. Patients infected with genotype IV exhibited a lower prevalence of anti-GOR antibody than patients infected with other genotypes.

Long-term follow-up of liver function in Italian haemophilic patients.

A 14-year follow-up of 153 haemophilic patients treated with clotted factor concentrates was made to assess liver function. Investigations included measurement of alanine amino transferase (ALT), immunoglobulins, HIV antibody, HBsAg, antibody to HBs and HBc antigens, antibodies to HCV (assayed by ELISA-2 and by RIBA-2) and HCV viraemia assessed by PCR. Assessment of liver function also included liver ultrasound scan and oesophageal X-ray. A positive correlation was found between abnormal liver function and age, HIV infection, HBV infection, HCV infection and hepatomegaly. However, there was no correlation between abnormal liver function and annual clotting factor concentrate use. The increasing incidence of liver insufficiency with increasing age is likely to result in substantial morbidity and mortality amongst haemophilic patients.

Hepatitis C virus serological and polymerase chain reactions in human immunodeficiency virus-positive and -negative patients.

BACKGROUND: Polytransfused patients may be dually infected with hepatitis C virus (HCV) and human immunodeficiency virus (HIV). OBJECTIVES: To assess the correlation of antibodies to HCV with viral RNA in serum as determined by polymerase chain reaction (PCR) in anti HIV-positive and -negative haemophiliacs. STUDY DESIGN: Serum from 150 Patients with or without HIV infection were examined for anti-HCV by second generation enzyme-linked immunosorbent assay (ELISA) and recombinant immunoblotting assay (RIBA). A sample was also tested in a nested-reverse transcription PCR for a conserved sequence of the 5' untranslated region of HCV. PCR-positive specimens were titrated and a type-specific PCR using viral core gene sequences was used to determine distribution of HCV viral types. RESULTS: Eighty-seven percent of the patients were positive in ELISA. All the positives but 2 were either positive of indeterminate in RIBA. The frequency of indeterminate RIBA results was 33% among HIV-positive subjects and less than 1% among HIV-negative ones. PCR was positive in 68% of 73 RIBA-positive or -indeterminate individuals and negative in all HCV-seronegative individuals examined. No significant differences were observed in HCV viral type, prevalence or titers of viraemia between HIV-positive or -negative patients. CONCLUSIONS: The majority (68%) of anti-HCV-positive haemophiliacs examined in this study had HCV RNA in their sera and anti-HCV profile determined by RIBA had no apparent influence on viraemia. The presence of HIV infection in these patients had no significant impact on HCV RNA prevalence, titer or HCV type distribution.

Prevalence of hepatitis C virus genotypes in Italy.

Hepatitis C viruses (HCV) present in 110 Italian patients were characterized by genotype-specific PCRs. Among the 65 cases of community-acquired hepatitis, HCV genotype II was dominant (60%), followed by genotypes IV (15%), III (11%), and I (3%). Among the 45 hemophilia-associated cases, the distribution of the four HCV genotypes was markedly different: genotype I was the most prevalent (61%), followed by genotypes II (25%), III (4%), and IV (2%). Double infections were observed in eight patients. Two HCV remained unclassified. For the 45 community-acquired cases from which a liver biopsy was available, genotype II was associated with more severe liver damage than the other types.

Hepatitis C virus seroprevalence in Italian haemophiliacs injected with virus-inactivated concentrates: five year follow-up and correlation with antibodies to other viruses.

The overall prevalence of anti-HCV antibody in a group of 125 haemophiliacs was 62%. Four patients who had never received replacement therapy were anti-HCV negative. Of the 121 patients injected regularly with commercial concentrates, 76 were already anti-HCV seropositive in 1985 and remained so throughout the follow-up. Two patients seroconverted in 1987 without obvious signs or symptoms of hepatitis. Our patients were treated with dry heat-treated concentrates since 1985 and with wet heat- or solvent/detergent-treated concentrates since 1988. The absence of further seroconversions and of symptoms of acute post-transfusion non-A, non-B hepatitis since 1988 suggest that present virucidal treatments of concentrates are effective in preventing HCV transmission. Anti-HCV positivity appeared to be unrelated to the type and degree of haemophilia as well as to the presence of antibodies to hepatitis B virus, human immunodeficiency virus type 1, and human herpesvirus type 6.

Gene deletion in an Italian haemophilia B subject.

DNA from 20 Italian haemophilia B patients was analysed by the Southern blotting technique and hybridisation to a factor IX cDNA probe. A large deletion of factor IX gene was detected in one patient with antibodies to the infused factor; the EcoRI pattern of the other 19 subjects examined was normal.

Standardization of factor VIII: establishment and use of secondary standards.

Two secondary standards for use in routine assays of Factor VIII in therapeutic concentrates and in patients, plasmas, respectively, have been established in a multicenter collaborative study. In order to assess the effect of the adoption of these preparations as common Secondary Standards a comparative assay has been performed: one sample of a Factor VIII concentrate of intermediate purity and one plasma sample have been tested in two laboratories for Factor VIII:C activity using as reference, among others, the common working standard. Analysis of the results shows that with the plasma sample the differences of the estimates obtained with any of the references in our two laboratories were not statistically significant (P greater than 0.3), while with the concentrate sample the differences were always statistically significant (P less than 0.005). The study shows that the adoption of common working standards (besides the uniformity in assay method, reagents and basic equipment) is not sufficient to eliminate interlaboratory variation in the measurement of Factor VIII:C.

Comprehensive study of haemostasis in chronic uraemia.

A comprehensive study of haemostasis has been performed in a homogeneous group of 25 adult patients with conservatively treated chronic uraemia. We have found prolonged bleeding time, impaired platelet adhesiveness and aggregation, and decreased platelet factor 3 activity, increased values of fibrinogen, of factor VIII activity and related antigen, and of combined levels of factors II, VII and X. Non-significantly abnormal concentrations of factor XIII and of plasminogen and significantly lower values of plasminogen activators and higher values of urokinase inhibitors and anti-plasmin have also been found. Fibrinogen degradation products were detected in the urine of some patients.

Comprehensive study of haemostasis in nephrotic syndrome.

A comprehensive study of haemostasis has been performed in a homogeneous group of 20 patients with nephrotic syndrome without renal failure. We have found unchanged number of platelets and a significant increase of platelet adhesiveness and aggregation; increased levels of activity and related antigen of fibrinogen, of factor VIII, of activity of factors II, VII and X and of antigens of factors XIII. Antithrombin III was unchanged in plasma and was detected in the urine. Euglobulin lysis times were decreased, and levels of plasminogen and its activators were increased after a venous occlusion test. At the same time urokinase inhibitors and antiplasmins were increased not only after, but also before a venous occlusion test. Fibrinogen degradation products have been found in the urine of all our patients but not in their sera.

[Chronic renal insufficiency. Low-protein diet supplemented with essential amino acids and ketoanalogues]. / Insufficienza renale cronica. Dieta ipoproteica supplementata con aminoacidi essenziali e chetoanaloghi.

Conventional conservative treatment of chronic uremia is aimed at reducing protein intake to a minimum level compatible with nitrogen balance. A great progress in this field is represented by a low protein diet associated with some essential aminoacids and ketoanalogues. A group of 20 patients with chronic renal failure at an advanced stage has been studied and good results have been obtained with a low protein diet (0.2 g/kg/24 h vegetable proteins) associated with essential aminoacids and ketoanalogues.

Characterization of heterogeneity of haemophilia B for the detection of carriers.

Factor IX activity (IX:C) and antigern (IX:Ag) as well as factor VII, prothrombin time, Normotest and Thrombotest were determined in 37 haemophiliacs B from 26 kindreds, in 30 carriers of haemophilia B and in 40 healthy subjects. In 15 obligatory carriers from 9 kindreds, in which haemophiliacs did not exhibit IX:Ag in excess of IX:C, IX:C was parallel with IX:Ag, but in 4 from 4 kindreds, in which haemophiliacs exhibit excess of IX:Ag, there was a discrepancy between IX:C and IX:Ag. Among the obligatory carriers 6 from 3 kindreds, in which haemophiliacs did not have IX:Ag in excess of IX:C, showed that the anomaly M is not a sign of genetic variability of haemophilia B.

Antithrombin III, heparin cofactor and antifactor Xa in relation to age, sex and pathological condition.

Antithrombin III (At-III) activity and protein, heparin cofactor activity, antifactor Xa activity were determined in 200 healthy adults, evenly distributed within age and sex groups, in 60 patients with cerebral thrombosis and in 20 oral contraceptive users. There was a positive correlation between At-III protein and its activities in normal subjects and in patients with cerebral thrombosis. The mean At-III protein and heparin cofactor activity values decreased with age in men and in women of fertile age. The mean values of At-III protein and its activities did not show any variation in the patients with cerebral thrombosis when compared with the normals. In oral contraceptive users a positive correlation was also found between At-III protein and its activities, antifactor Xa activity excepted. The mean antifactor Xa activity value in these women decreased during treatment, whereas the other At-III activities and At-III protein were unchanged.

[Cutaneous iontophoresis of hyaluronidase in the treatment of hemorrhages in hemophilic patients]. / L'iontophorèse à la hyaluronidase par voie cutanée dans le traitement des hémorragies chez les sujets hémophiles.

After a surgery of the theoretical background of iontophoretic technic and the physico-chemical properties of hyaluronidases, a clinical application is presented, based on over 500 hemophilic patients treated percutaneously, in cases both of hemarthrosis and hematomas. The results are presented and discussed. The conclusion is that hyaluronidase iontophoresis is useful and should be performed together with the other measures for the prevention and the treatment of the damages produced by hemorrhages in hemophilic patients.