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Attempts to differential hepatitis B from hepatitis A infection by newly developed serological tests.

An immune Indian ink micro-agglutination method has been evolved for the detection of an antigen present in the blood associated with infectious hepatitis (called IHxAg). In previous studies 86% of serum samples taken from children with hepatitis A proved to be positive by this technique. Present studies were related to 239 adult in-patients with a clinical diagnosis of hepatitis A (123 cases) or hepatitis B (116 cases). Blood samples taken serially during the illness were tested for IHxAg, HBsAg and anti-HBsAg. The results were in accordance with the clinical diagnosis in 60% in contradiction in 30%, whilst all tests brought negative results in 10%. The clinical laboratory findings (SGPT, thymol turbidity) were more in harmony with our laboratory results than with the clinical diagnoses. Rheumatoid factor did not disturb the immune Indian ink reaction, labile serum proteins caused, however, non-specific reaction in 30% of serum samples. When durocytes were used instead of Indian ink the rate of false positive results dropped to 10%. Sera taken in convalescent phase from patients with IHx antigenemia in the acute phase of illness contained an antibody against IHxAg. A crude gammaglobulin preparation from a pool of convalescent sera gave a precipitation line in agarose gel with an antigen present in the fecal extract of children with hepatitis A. This precipitation line proved to contain virus-like particles with an approximate diameter of 25 nm when tested by electronmicroscopy. No precipitation could be seen when sera of the same patients were tested against the same gammaglobulin preparation.

Adult

Transmission of non-A, non-B hepatitis.

In studies conducted in the early 1950s, sera from six asymptomatic blood donors, implicated in the transmission of viral hepatitis, were inoculated into 10 to 20 volunteers each. Five of these "implicated" donor sera transmitted clinically apparent hepatitis to the recipients. The stored serum samples from these studies have been reanalyzed using serologic markers for hepatitis B virus and hepatitis A virus infection. Two of the donor sera were hepatitis B surface antigen (HBsAg)-positive, and both transmitted hepatitis B virus infection to all susceptible recipients, half of whom showing clinical symptoms. The remaining three infectious donors were HBs-Ag-negative, yet were icterogenic to 10% to 47% of recipients. Testing of serum samples from these recipients with hepatitis showed no evidence of hepatitis B virus or hepatitis A virus infection. This study and other recent evidence suggest that there is a third type of human viral hepatitis--non-A, non-B hepatitis--which is due to a transmissible agent and may well be associated with a chronic carrier state.

Adult

Risk factors in transmission of non-A, non-B posttransfusion hepatitis. The role of hepatitis B antibody in donor blood.

Risk of developing icteric hepatitis in a transfusion study involving cardiac surgery patients was 0.2 per cent per unit of blood transfused with the ratio of icteric to anicteric cases being 1:4. Risk of developing hepatitis was proportional to the number of units transfused: one to four units, 4 per cent; six to ten units, 8 per cent; 11 to 20 units, 19 per cent; greater than 21 units, 42 per cent. The prevalence of type B hepatitis was low (6 per cent), with the vast majority of patients being shown to have non-A non-B hepatitis. However, a greater incidence of hepatitis type B serologic events was observed among recipients of anti-HBs positive blood than those transfused only with units not containing antibody (p = 0.04. A significantly greater incidence of non-A, non-B hepatitis was observed among patients transfused with blood containing anti-HBs when compared with a group who received blood without antibody (p less than 0.01). Caution should be exercised in interpretation of this difference because patients transfused with blood containing anti-HBs received significantly more units of blood. However, utilization of stepwise regression analysis to unconfound the two dependent variables suggest that the use of blood containing anti-HBs increases the hepatitis risk (p = 0.06) although the number of units transfused was the more significant factor (p less than 0.001). Additional data from carefully designed studies are needed to determine if donor blood containing anti-HBs significantly increases the risk of transmitting non-A, non-B hepatitis.

Antibodies, Viral

[Change of the prevalent subtype of hepatitis B antigen in acute hepatitis B infections (author's transl)].

Subtyping of hepatitis B antigen (HBs Ag) in patients with acute hepatitis B revealed subtype ay in 75 percent while subtype ad was much more common in chronic hepatitis B infections: 81 percent of HBs Ag positive blood donors and 76 percent of patients with HBs Ag positive chronic aggressive hepatitis revealed subtype ad. In contrast to blood donors and chronic hepatitis patients, a change of the prevalent subtype was noted in acute hepatitis patients between 1970 and 1974. Before May 1972, subtype ad was found in 67 percent of patients, whereas later subtype ay dominated in 93 percent. An unequal distribution of subtypes between acute and chronic forms of hepatitis B infections has been explained by differences in the virulence of virus strains. Our results suggest that the higher prevalence of subtype ad in longterm carriers may be due to infection during an earlier time when subtype ad was also common in acute infections. The change of the prevalent subtype in acute infections may be attributed to differences in contagiosity rather than differences in virulence of virus strains.

Acute Disease

Antibodies to hepatitis B core antigen in hepatitis B surface antigen-positive and -negative chronic hepatitis.

Antibody to hepatitis B core antigen (anti-HBc) is a sensitive indicator of hepatitis B virus (HBV) infection. However, anti-HBc has been found in only a few patients with chronic hepatitis. Therefore, we tested for anti-HBc in 124 sera from 67 patients with histologically proven chronic hepatitis by the indirect fluorescent antibody technique. All patients, except for one with chronic hepatitis who was seropositive for hepatitis B surface antigen (HBs Ag), had anti-HBc that persisted throughout the follow-up period (three months to three years). Of 33 HBs Ag-seronegative patients, anti-HBc was detected in seven patients and persisted for six months to two years. These findings suggest that in this study 21% of patients with chronic hepatitis with undetectable amounts of HBs Ag in the serum had evidence of recent or continued HBV replication.

Adolescent

Prevention of hepatitis B in hemodialysis patients using hepatitis B immunoglobulin. A controlled study.

In a controlled study, the protection effect of hepatitis B immune globulin (HBIG) was evaluated in patients hemodialyzed for less than one month in two collaborating units. Fifteen randomly selected patients received HBIG at five to eight week intervals throughout the study, and 13 other control patients received no immunoglobulin. During a follow-up period of 14 to 30 months, none of the HBIG-treated and 12 of the control patients developed evidence of exposure to virus B hepatitis, including 10 with HBs Ag antigenemia (p is less than 0.001): five of these remained persistently antigen positive. Evidence of non-B hepatitis was found in 8 HBIG-treated and in 3 non-treated patients. Only two HBIG-treated patients developed active antibodies against hepatitis B surface antigen. Thus, HBIG seems effective in preventing hepatitis B in hemodialysis patients, provided the interval between two injections is not greater than two months. However, prolonged administration of HBIG may impair passive-active immunization to hepatitis B virus.

Adult

Transmission of hepatitis B to chimpanzees by hepatitis B surface antigen-positive saliva and semen.

To assess the infectivity of hepatitis B surface antigen (HBsAg)-containing body fluids other than blood, chimpanzees were inoculated intravenously with saliva and semen obtained from HBsAg-positive individuals implicated in non-percutaneous transmission of hepatitis B. Saliva and semen samples were negative for occult blood. The titer of HBsAg in saliva was on the average only 1/3,000 that of the corresponding serum. One chimpanzee, inoculated sequentially with saliva from three individuals, developed HBsAg at 9 weeks and serum glutamic pyruvic transaminase elevation at 13 weeks after injection. HBsAg persisted for 15 weeks. This animal also developed e antigen, anti-core antibody, and anti-surface antibody. Liver biopsies showed acute hepatitis that subsequently resolved. A second chimpanzee, inoculated with HBsAg-positive semen, developed HBsAg and elevated serum glutamic pyruvic transaminase 4 weeks after inoculation and then died suddenly without explanation. HBsAg was positive in two consecutive samples and was confirmed by specific neutralization. Autopsy did not reveal evidence of hepatitis. This study demonstrates that HBsAg-positive saliva and, probably, semen contain infectious virus and suggests that saliva and/or semen may serve as important mechanisms in the transmission of type B hepatitis.

Animals

Type B hepatitis in Iran.

Hepatitis B surface antigen (HBsAg) was found in 1% of controls, 2.1% of professional blood donors, 2.0% of leprosy patients and 76.1% of acute hepatitis in Tehran and Mashhad, Iran. All HBsAg positive samples also possessed antibody to the hepatitis B core antigen and all were subtype ayw. Type B hepatitis and the HBsAg state are frequent in Iran, but most must be accounted for by "non-parenteral" or inapparent" parenteral exposure.

Acute Disease

Hepatitis B virus infections among Danish dentists.

Since type B hepatitis is generally regarded as an occupational risk for dentists, the participants at the 1976 annual meeting of the Danish Dental Association were examined for hepatitis B surface antigen (HBsAg) and antibody to HBsAg (anti-HBs). A total of 1,338 dentists (89% of the dentists at the meeting and 29% of all Danish dentists) were included in the study by completion of a questionnaire and by radioimmunoassay of a blood sample for HGsAg and anti-HBs. None of the dentists was HBsAg-positive, but 110 (8.2%) had anti-HBs. An increasing frequency of anti-HBs was found with increasing age, but the figures were similar to the findings in a control population. Evidence is presented that hepatitis found before admittance to or during the time at dental school was predominantly not of type B. In contrast, type B hepatitis predominated during the professional activity of the dentists. On the basis of the serological findings in 29% of all Danish dentists, it is concluded that dentists cannot be regarded as a high-risk group for hepatitis B.

Aging

Dane particles and associated DNA-polymerase activity in saliva of chronic hepatitis B carriers.

To investigate furhter the problem of salivary transmission of type B hepatitis, salivas free of blood contamination from three HBsAg-positive carriers with chronic active hepatitis were examined by CsCl equilibrium density gradients and electron microscopy (EM). In the CsCl gradient HBsAg of whole salivas distributed in a band centered at 1.19gm/cm3 with a clearly defined shoulder at 1.24 gm/cm3; the HBsAg was found mainly in the mucous component. On EM examination, fractions from the 1.19 gm/cm3 peak contained spherical HBsAg particles of 22 +/- 3 nm diameter, whereas in the 1.24 gm/cm3 shoulder Dane particles 43 nm in diameter with 28 nm cores were found. Specific hepatitis B virus associated DNA-polymerase activity also was found in the 1.24 gm/cm3 shoulder where the Dane particles occurred and was absent from the saliva of healthy controls. When salivas were incubated for three hours at 37 degrees C the total amount of HBsAg diminished and the 1.24 gm/cm3 shoulder disappeared, probably as a result of endogenous degradation of the Dane particles and the free HBsAg. These findings clearly indicate that the hepatitis B viral particle is present in the saliva of chronic HBsAg carriers with active disease and further confirm that saliva is an important vehicle of infection.

Adolescent

Correlation of e antigen, DNA polymerase activity, and Dane particles in chronic benign and chronic active type B hepatitis infections.

The e determinant of hepatitis B surface antigen (HBS Ag) was found in 23 of 42 patients with chronic hepatitis B virus (HBV) infection. Presence of e antigen was associated with increases in DNA polymerase activity and in the number of circulating Dane particles. In the group with detectable e antigen, the average DNA polymerase activity was 367+/-78 counts per minute (cpm; mean+/-standard error [SE]), and the average number of Dane particles counted in electron micrographs was 4.4% of the total HBS Ag. In contrast, e antigen-negative patients had an average DNA polymerase activity of 40+/-6.9 cpm (P less than 0.1) and an average Dane particle count equal to 0.6% of the HBS Ag. The e antigen was detected in 68% of patients who were HBS Ag carriers or had persistent viral hepatitis and 40% of those with chronic active type B hepatitis. Thus, the presence of e antigen correlated with both the chronicity and presence of infectious HBV. However, it did not correlate with the type or severity of liver disease after HBV infection, since e antigen was present in both chronic benign and chronic aggressive hepatitis B infections.

Adult

Full and empty Dane particles in chronic hepatitis B virus infection: relation to hepatitis B e antigen and presence of liver damage.

Circulating complete and defective hepatitis B virus forms, as represented by full, DNA polymerase-positive and empty, DNA polymerase-negative Dane particles, respectively, were investigated in sera from patients with chronic hepatitis B virus infection and related to the presence of e antigen and antibody and to the histological findings on liver biopsy. Complete hepatitis B virus particles were detected in the serum of all patients postive for e antigen, their percentage ranging from 15 to 61% of the total Dane particle population. Although most of these cases had chronic persistent or chronic active hepatitis, complete viral particles were also found in serum of 3 healthy carriers of hepatitis B surface antigen who had e antigen. These results indicate that e antigen is a marker of active virus replication and support its association with infectivity. It is also associated with liver damage because production of complete virus is a feature of chronic hepatitis. In the presence of anti-e, detection of Dane particles in serum appeared to be related to the histological findings. Most of the healthy carriers had no Dane particles in serum, whereas 80% of the cases with chronic liver disease had circulating Dane particles. However, in contrast to the cases with e antigen, 98 to 100% of Dane particles in these cases appeared to be defective in nucleic acid material on electron microscopy after positive staining. All of the patients with chronic active hepatitis in this group had progressed to cirrhosis and it is possible that production of complete virus particles is reduced in the later stages of the illness.

Carrier State

Papular acrodermatitis of childhood and hepatitis B infection.

A case of papular acrodermatitis (PAC) associated with acute anicteric type B hepatitis occurred in a 2-year-old child. Immunocytochemical studies failed to detect the presence of viral antigens in the involved skin lesion. Current knowledge of the hepatitis B viral antigens and of their possible role in PAC is discussed.

Acrodermatitis