Detection of hepatitis B virus DNA in serum using synthetic non-radioactive oligonucleotides.

A rapid and simplified technique for detecting hepatitis B virus (HBV) DNA by spot hybridisation in the sera of patients with different clinical forms of HBV infection was investigated using enzyme conjugated synthetic oligodeoxyribonucleotides as probes. These are able to hybridize to the S and C regions of the HBV L(-) DNA strand. When compared with a complete 32P-labelled HBV DNA probe, the synthetic oligonucleotides provided a sensitive and quick method for the routine survey of HBV infection. Moreover, the DNA extraction procedure used allowed the spot hybridisation technique to be applied and read easily and the results obtained within a few hours. It is concluded that synthetic cold probes can be used in hybridisation assays HBV DNA detection as part of current clinical laboratory procedures.

Complement-fixation test for rotavirus detection: comparison and analysis of different methods to reduce anti-complementary activity of some specimens.

The complement-fixation test may be used to detect rotaviral antigens directly in clinical specimens. However, a certain number of specimens tested for human rotaviruses by the complement-fixation test show an anti-complementary activity. By comparing eight techniques we analysed this anti-complementary activity and identified the best method for its reduction. Pretreatment of clarified supernatant of stool suspensions by some methods resulted in a reduction of anti-complementary activity, without reducing the sensitivity of the method. Clarified supernatants of 8/36 (22.2%) specimens were anti-complementary; this anti-complementary activity was best removed by absorption with fetal calf serum or calf albumin. Such treatment offers practical means of increasing the specificity of complement-fixation test. Some observations suggest that the anti-complementary activity of stool suspensions may be frequently due to presence of one or more chelating agents that may be in faecal specimens.

An unusual spinal presentation of Whipple disease.

SUMMARY: When Whipple disease (WD) is confined to the CNS, diagnosis may be difficult. We report a case of WD with spinal presentation in an otherwise healthy woman who had a 5-year history of relapsing-remitting cervico-thoracic myelopathy. We suggest that the diagnosis of WD should be considered in the presence of an enlarged and enhancing spinal cord even in the absence of any systemic involvement.

Multicenter evaluation of the new HIV DUO assay for simultaneous detection of HIV antibodies and p24 antigen.

A multicenter survey was performed to evaluate a new semi-automated human immunodeficiency virus fourth generation antibodies and antigen simultaneous assay. This assay showed a sensitivity of 100% and specificity of 99.6% among sera obtained from hospitalized patients or blood donors. Sera obtained from commercially available as well as in-house seroconversions were tested showing that HIV DUO is able to reveal an infected state in 11 out of 14 cases earlier than conventional tests. This new assay improves old test performances in terms of sensitivity, maintaining specificity at very high levels.

Further considerations on the changing face of pneumonia in Marche Region (central Adriatic coast of Italy).

The present paper provides a more detailed review of some themes already discussed in previous papers concerning the changing face of pneumonia in an area of the central Adriatic coast of Italy, including an ex novo examination of the correlations between radiological findings and serological groups not previously reported. The discussion concentrates particularly on some questions raised by the research, such as the apparent rarity of pneumococcus, the elevated number of seronegatives appearing as atypical pneumonia, the high percentage of "bacterial-type" cases occurring among the viral-like, and the therapeutic problem.

[The new aspect of pneumonia in an area of the Middle Adriatic]. / Il nuovo volto delle polmoniti in un'area del medio Adriatico.

The present work reports the findings of a ten-year, research, the aim of which is to outline current views of pneumonia in a zone of the Middle Adriatic (USL 17-Regione Marche). The study confirmed, in its entirety, a lesser aggressiveness and loss of the "seasonal feature" of today's pneumonias. The high incidence of Atypical Pneumonia (AP) due to viral-like microorganisms (mycoplasma, chlamydia, coxiella, legionella) and an even higher one of unknown etiology is reported. After having analysed the most likely reasons for such a change and its various implications, the authors conclude that this type of research should be extended in order to trace a map of the more common infectious agents in single geographical zones, as an indispensable premise for a more concrete etiological diagnosis and for a more rational choice of the antibiotic.

Induced underglycosylation of PLC/PRF/5 human hepatoma cells. Brief report.

The production of HBsAg and human plasma proteins by PLC/PRF/5 human hepatoma cell line was studied in the presence of glycosylation inhibitors. The data presented here suggest that HBsAg release in culture medium by these cells is not reduced by an inhibition of glycosylation and that Tunicamycin may play a role in a mobilization of the intracellular pool of antigen.

Modulation of production of hepatitis B surface antigen by a human hepatoma cell line.

Three drugs were assayed for their capacity to inhibit hepatitis B surface antigen (HBsAg) production by the PLC/PRF/5 human hepatoma cell line. The effect on cell growth and HBsAg production of Cordycepin, 6-azauridine, and Hygromicin B is reported. Hygromicin B, a translation inhibitor unable to penetrate normal cells, greatly reduced HBsAg production by growing and confluent cells.

Comparative effects of alpha, beta and gamma human interferons on a HBsAg-producing human hepatoma cell line.

The effects of all the three types of human interferons (alpha, beta and gamma) on a human hepatoma cell line with hepatitis B virus (HBV)-DNA sequences integrated into the host DNA and producing hepatitis B surface antigen (HBsAg) in culture medium were assayed. The aim of the present research was to test human interferon preparations in an in vitro system for hepatitis B virus, and to compare the observed effects. The results evidenced both the antireplicative activity principally showed by preparations of beta and gamma human interferons and the inhibition of HBsAg production by high concentrations of gamma human interferon.

Psittacosis in a highly endemic area in Italy.

In one locality in Italy where the incidence of psittacosis has increased rapidly since 1980, a hospital-based study and a seroepidemiological survey were carried out in order to define the clinical and epidemiological features of psittacosis in that area. Registers of the Virology Unit of the University of Ancona, Italy, were reviewed and all hospitalized patients with a serological diagnosis of psittacosis were identified. A total of 76 cases were found and studied. A presumptive bird source was identified in 80% of 62 patients, on whom a detailed investigation had been possible. Poultry represented the most frequent probable source of infection. Clinically, the predominant pattern of illness was a moderately severe lower respiratory tract infection, with chest X-rays showing pulmonary shadowings in 68 patients (89%). In the seroepidemiological study, 51 out of 143 subjects were exposed to birds (35.7%), but only 7 out of 96 urban adult blood donors (7.3%) were positive for chlamydial antibodies using the microimmunofluorescence test.

[Cross-over electrophoresis on cellulose acetate. A new method for rapid diagnosis of rotavirus infection]. / Crossing-over elettroforesi su acetato di cellulosa. Una nuova applicazione alla diagnosi rapida delle infezioni da Rotavirus.

A new application of crossing-over electrophoresis on cellulose acetate (c.o.e.) in diagnosis of Rotavirus infections is described. The results are compared with those obtained in electron microscopy (e.m.). The main reasons for application of c.o.e. to routine virological diagnosis of Rotavirus infections are exposed.

[Pulmonitis caused by Coxiella burneti and Chlamydia psittaci. 2 years of research in Marche]. / Polmoniti da Coxiella burneti e Chlamydia psittaci. Due anni di ricerche nelle Marche.

This paper point to infections caused by Chlamydia psittaci and Coxiella burneti in a region of the middle Italy (Marche) from April 1981 to March 1983. 32 of 686 cases (about 5%) of pneumonia were diagnosed as Q Fever or Ornithosis-Psittacosis. The role of the research of these etiologies in routine diagnostic work on acute respiratory infections is underlined.

Diagnosis of human rotavirus infections: comparison of an electrophoretic method, a modified complement fixation test and electron microscopy for rotavirus detection.

A modified complement fixation test and a counter-immuno-electrophoresis on cellulose acetate were compared to electron microscopy, in detecting Rotavirus from stool specimens. Out of 75 samples from patients with acute gastroenteritis, 40 per cent yielded positive results. The main advantages of the above two methods in routine diagnosis are discussed.

Insulin reduces HBsAg production by PLC/PRF/5 human hepatoma cell line. Brief report.

Although its action at the molecular level is not completely understood, insulin, as well as its antagonist glucagon, certainly plays an important role in the modulation of protein synthesis. In order to observe whether insulin is involved in virus gene expression, we studied its effect on PLC/PRF/5 human hepatoma cell line, which posses HBV DNA sequences integrated at several sites. While human insulin had no effect on cell growth and increased the production of two plasma proteins, a selective inhibitory effect on HBsAg production could be detected. This observation might be useful for further studies both on virus gene expression and insulin action at the molecular level.

HBV infection in hemodialysis patients: monitoring and prevention.

From 1986 to 91, 174 dialysis patients were studied. The prevalence of previous HBV infection and of chronic carriers was 33.3 and 4.6%, respectively. Immunization rate after vaccination (3 doses) was 63%. In 1991, we proposed a vaccination with 4 doses and recommend a 6-monthly anti-HBs evaluation to assess the timing of any booster dose needed.

Risk factors and clinical expression of HCV infection in hemodialysis patients.

In our Dialytic Unit, the prevalence of antibodies to HCV (anti-HCV) was 16.4% in 1990 (73 hemodialyzed) and 21% in 1991 (105 hemodialyzed). The incidence of seroconversion was 13.1%. The transaminase behavior was similar in a group of anti-HCV-positive and a group of anti-HCV-negative patients and with previous HBV infection. Blood transfusions and dialytic age did not affect anti-HCV positivity.

[Observations about the treatment of Cytomegalovirus retinitis in patients with HIV infection using Ganciclovir and Foscarnet] / Osservazioni relative al trattamento con Ganciclovir e Foscarnet della retinite da Cytomegalovirus in pazienti con infezione da HIV.

Objective: The Authors report their experience of Cytomegalovirus (CMV) retinitis therapy in HIV patients, using Ganciclovir and Foscarnet in monotherapy. They also evaluate the reliability of the Polymerase Chain Reaction (PCR) through the qualitative technique as an index of active disease. Methods: 18 patients suffering from CMV retinitis were treated: Ganciclovir was administered at a dosage of 10 mg/kg b.w./day and Foscarnet at 180 mg/kg b.w./day, both of them for 21 days during the induction phase. During the mantainance phase the former was administered at 5 mg/kg b.w./day and the latter at 90 mg/kg b.w./day for 5 days a week. Results: Both the drugs induced the stabilization or regression of the lesions. There was however a relapse with both therapies. We did not observe a significant difference either in the entity and the duration of the stabilization or in the survival from diagnosis time. Finally the PCR method was not helpful in the diagnosis of CMV retinitis.