Comparison between reverse transcription-polymerase chain reaction and immune enzyme assays in the diagnosis of hepatitis C virus infection.

This is the first study of HCV infection performed in Romania by reverse transcription-polymerase chain reaction (RT-PCR). We set up our own protocol, using "nested" primers located in the 5' UTR of HCV. Thirty two patients, 16 with chronic hepatitis, 7 with hepatic cirrhosis and 9 without chronic hepatic disease were investigated. The assignment of each patient to one of the groups was based on clinical and laboratory criteria, especially transaminase levels and hepatic biopsy findings. In 14 out of 16 (87.5%) chronic hepatitis cases and in 6 cases of hepatic cirrhosis out of 7 (85.7%), there was good agreement between RT-PCR and ELISA results. Transaminase levels were concordant with RT-PCR results in 27 of 29 cases (93.1%) but in only 21 of 29 cases (72.4%) transaminase levels were in agreement with ELISA results. In two patients treated with interferon, transaminase levels and the other clinical and laboratory parameters fell in the normal range in parallel with the disappearance of viremia, whereas anti-HCV antibodies were still detectable. RT-PCR has a higher specificity in detecting the etiology of hepatic diseases if compared to ELISA and an at least equal sensitivity. The method proves to be the best means for the confirmation of HCV infection, and for the monitoring of interferon therapy.

Serological profile assessment of the infection with hepatitis C virus (HCV) in haemophiliacs and thalassemic patients.

Polytransfused patients represent a major risk group for hepatitis C (HCV) acquirement. Haemophiliacs and thalassemic patients treated with virus contaminated blood or blood derivatives frequently exhibit anti-HCV antibodies and signs of chronic hepatitis. The serological profile for the HCV infection was investigated in 13 haemophiliacs, 18 cases of thalassemia and in 14 polytransfused patients affected by other diseases. The presence of anti-HCV antibodies was detected by means of the ORTHO HCV 3.0 ELISA kit and confirmed by Western-blot Murex. The serotyping used synthetic peptides mimicking the immunodominant epitopes in the NS4 region, characteristic of each of the six HCV genotypes in an ELISA blocking reaction (Murex). Serotype 1 was prevalent (51.1%), while serotype 2 was detected in 13.3% of patients, with a higher frequency in thalassemia cases. The remaining samples were multireactive, and serotype 3 alone was not detected. The profile of Western-blot bands was distinct for the monoreactive samples belonging to serotype 1 or 2. The analysis of the multireactive samples in young (thalassemic, age mean 15.17 +/- 6.5) and aged patients (haemophiliacs, age mean 32.64 +/- 13.5) allows us to suggest a different succession of reinfection acquirements. The infection with one of the subtypes does not confer protection against the reinfection with others. However, a certain attenuation of the symptomatology is obvious in the case of reinfections, indicating the existence of crossantigenic reactivities which contribute to protection. This protection is more evident in the case of primary infection with type 2 and is partially due to antigens coded by the NS4 genomic segment.

Immunological disorders of increased severity in children with AIDS associated with hepatitis B and C infections.

Two groups of children were set up: children infected with HIV and hepatitis viruses and children infected with HIV, but not with hepatitis viruses, too. The immunological aspects investigated referred to the Ig serum value, the absolute number of T CD4 lymphocytes and the T CD4/T CD8 ratio. The subjects of the first group (in whom hepatitis markers were present) displayed increased IgA and IgM values at a higher rate than those of the second group (54.87% against 32%, 83.3% against 53%), as well as a lowering below 300/ml of the number of T CD4 lymphocytes (50% against 29.4%). On the other hand, increased IgG levels and values below 0.8 of the T CD4/T CD8 ratio were found at similar rates in the two groups of children (77.4% against 80% and 70.5% against 70% respectively). By means of the data obtained, the authors try to point out one of the ways by which hepatitis viruses, considered as a potential cofactor in the AIDS development, contribute to the course of this disease, namely by intensifying the immunological disorders.

Serological diagnosis of chronic viral hepatitis.

Chronic viral hepatitis must be distinguished from other chronic liver disorders. The clinical picture associated with typical biochemical findings provides no definite information about the causative agent. An important factor, both for the patient and for the individuals in contact with him, is the evaluation of his infectivity. From 172 chronic hepatitis disorders the serological diagnosis confirmed the viral etiology in 120 patients (69.7%). HBsAg was present in 33.7% (with HBeAg in 5%), anti-HCV antibodies 22.6% and markers for both viruses in 13.4% of cases. The HCV etiology was more frequently found in chronic hepatitis (44.9%) in liver cirrhosis (50.7%). The smaller number of patients displaying simultaneously anti-HBs and anti HCV antibodies did not confirm the recent suggestions that HCV would be the most important hepatotropic virus which enhances HBsAg clearance in chronic hepatitis.

Considerations about the prophylactic role of T cell vaccination in HIV infection.

The AIDS epidemic in Romania has properties that are both common to countries of Eastern Europe and unique to Romania. The unique aspects include the large number of cases in children infected parenterally and a relatively low but steadily increasing number of HIV infected adults. Other groups of children and adults were also exposed at risk to acquire HIV by transfusion with unscreened blood or by multiple parenteral treatments with potentially contaminated needles. Our hypothesis was that a substantial number of people in area of high endemicity was "silently" infected but did not undergo specific immune response or clinical signs. In an attempt to explore the possibility that immunization with autologous proteins (consequence of multiple transfusions) can play a role in the protection from HIV infection or in slowing down the progression of disease, we compared the incidence of autoantibodies (anti-nuclear, smooth muscle and anti-HEp-2) in a group of long survivors children with AIDS, in HIV infected children with rapid evolution and in adults at risk for parenteral acquisition of blood borne viral disease. We analyzed also the incidence of some blood borne viral infections and the presence of anti-HLA antibodies in the same groups of patients. Our results do not support the suggestion that HIV infection or AIDS can be prevented, respectively, delayed by T-cell vaccination.

[A seroprevalence study of the hepatitis C virus among multiply transfused patients]. / Etudes sur la séroprévalence du virus de l'hépatite C chez les polytransfusés.

A study was conducted on 61 polytransfused patients, 24 patients submitted to long lasting parenteral treatments, 116 blood donors and 132 controls. Immuno-enzymatic tests were used for detection of antibodies against hepatitis C virus (HCV). They were found in 56 (91.8%) out of the 61 polytransfused patients, in 5 (4.3%) of the donors, in 5 (20.8%) of the patients with multiple treatments and in 5 (3.7%) controls. The VHB markers were detected at variable rates in all investigated groups. The high rate of seropositivity indicates that hepatitis C is becoming an important public health problem in Romania.

[Seroprevalence of hepatitis C virus among the multiply transfused]. / Etudes sur la séroprévalence du virus de l'hépatite C chez les polytransfusés.

A study was conducted on 61 polytransfused patients, 24 patients submitted to long lasting parenteral treatments, 116 blood donors and 132 controls. Immuno-enzymatic tests were used for detection of antibodies against hepatitis C virus (HCV). They were found in 56 (91.8%) out of the 61 polytransfused patients, in 5 (4.3%) of the donors, in 5 (20.8%) of the patients with multiple treatments and in 5 (3.7%) controls. The VHB markers were detected at variable rates in all investigated groups. The high rate of seropositivity indicates that hepatitis C is becoming an important public health problem in Romania.

The evolution of hepatitis B virus infection in children with symptomatic AIDS.

Hepatitis B virus (HBV) markers were determined in 80 children under 5 years of age with HIV symptomatic infection. Because of high carrier rate of hepatitis B virus in Romania we investigated as control a group of age matched 36 HIV negative children offsprings of HBsAg carrier mothers. Serological and epidemiological investigations in families of HIV infected children support horizontal nosocomial and not vertical transmission for HIV in contrast with HBV whose perinatal transmission can not be excluded. Concerning the probable route of HBV infection both groups of children seem to have a comparable risk for parenteral, contact-associated or maternal-neonatal transmission. HBsAg was detected in 76.25% HIV positive subjects and in 13.9% of control (P = 0.05). From all serum samples tested, only 12, all from the control group, did not present any markers of past or current HBV infection. Two serum markers have been used as an index of active HBV replication: HBe antigen detection and HBs antigen quantification in one or paired serum specimens. HBeAg was detectable in 20% of HIV infected children and only in 2.8% controls (P = 0.05). Almost all HBeAg positive patients have higher values for HBs antigenemia. HBsAg concentrations well above the assay cut off value (sample/cut off ratio > 15) were generally representative for HIV infected children (54% versus 5.6% in controls). The prevalence of hepatitis Delta markers and anti-HCV antibodies was not significantly higher in HIV infected children in spite of the fact that they are potentially exposed to a wider range of antigens.(ABSTRACT TRUNCATED AT 250 WORDS)

Virological investigations in adults with acute pneumonia.

Virological investigations (immunofluorescence reactions and isolation attempts with pharyngeal exudate specimens, as well as serological tests) were performed in 110 adult patients with pneumonia. Viral or inframicrobial agents were involved in 70 (63.7%) of the cases, either alone (27 cases) or in association with bacteria (43 cases). Parainfluenza and adenoviruses were most frequently encountered both in the cases with mixed (viral + bacterial) and in those with strictly viral pneumonia. Mycoplasma pneumoniae accounted for 11% of the cases; the role of chlamydial and rickettsial germs was insignificant.

Creutzfeldt-Jakob disease: experimental transmission in the guinea pig.

An experimental disease could be serially propagated in the guinea pig by intracerebral inoculation of cerebrospinal fluid or brain suspensions from 8 patients with Creutzfeldt-Jakob disease (CJD). The disease, characterized by tremor, ataxia and convulsions, appeared after a long incubation period (370-420 days). Microscopic lesions and electron optic features were similar to those described in human CJD and in experimental CJD in the chimpanzee.

Laboratory investigations in Balkan endemic nephropathy.

Serum samples from three patients with Balkan endemic nephropathy (BEN) were inoculated intraperitoneally to guinea pigs. After a very long incubation period (58-273 days) the animals developed an experimental disease characterized by apathy, tremor, convulsions and a fatal outcome. The disease could be serially propagated, the same clinical symptoms being recorded at each of the three passages performed. The morphological features of the experimental disease included glomerular and tubular lesions and the presence of interstitial fibrous and lymphoplasmocytic reactions. Different hypotheses on the etiology of BEN are discussed.

Serological investigations concerning the presence of antibodies to arboviruses in wild birds.

Serum samples from 8 species of migratory birds (Ardea cinerea, Plegadis falcinellus, Anas querquedula, Anser albifrons, Gavia arctica, Fulica atra, Phalacrocorax carbo, Larus minutus) were tested for the presence of hemagglutination-inhibiting (HAI) antibodies to alpha- and flaviviruses. HAI antibodies to alphaviruses (eastern equine encephalitis, western equine encephalitis, Sindbis, Middelburg, Semliki Forest) ranging in titer from 1/20 to 1/160 were detected in 46 serum samples; 22 serum samples gave positive reactions (titers: 1/20 - 1/80) to flaviviruses (West Nile, Ntaya). In certain cases antibodies to several antigens could be made evident in the same serum sample. The serological results are discussed in the light of the birds' migration pattern.

Further data on experimental Creutzfeldt--Jakob disease (spongiform encephalopathy) in the guinea pig.

Intracerebral inoculation to guinea pigs of cerebrospinal fluid or brain suspensions from 6 patients with Creutzfeldt--Jakob disease was followed in 4 cases by the appearance, after a long incubation period, of a serially transmissible experimental disease. The disease was characterized by tremor, discrete ataxia and convulsions and led to a fatal outcome within several days. Microscopic lesions in the brain of guinea pigs with the experimental disease consisted in glial proliferation, spongiform change and vacuolation of neuron cytoplasm. Electron microscopy proved that the vacuoles were situated in the endoplasmic reticulum and in the cytoplasmic processes.

Endemic Balkan nephropathy, a slow virus disease?

Serum samples from patients with endemic Balkan nephropathy (EBN) were inoculated intraperitoneally to four species of laboratory animals. It was only in the guinea pig that a serially transmissible experimental disease occurred, after a long incubation period. The morphopathological features in the guinea pig were similar to those encountered in the terminal phase of human EBN. The hypothesis that EBN might be a slow virus disease is discussed.

Laboratory data on the experimental transmission of Creutzfeldt--Jakob disease in the guinea pig.

Cerebrospinal fluid samples collected from 3 patients with Creutzfeldt-Jakob disease were inoculated intracerebrally to guinea pigs. After an incubation period of 4-14 months the animals developed a serially transmissible disease, characterized by loss of appetite, loss of weight, difficulty in movement, myoclonic convulsions and a fatal outcome within several days. The microscopic lesions caused by the experimental disease consisted in glial proliferation, spongiform change and vacuolation of the cytoplasm of proximal dendritic regions.

Effects of DDAVP in cranial diabetes insipidus as compared to other antidiuretic drugs.

Five antidiuretic drugs were administered in each of twenty patients with cranial diabetes insipidus (DI). A daily intranasal dose of 10 microgram DDAVP (Adiuretin) produced longer and stronger antidiuretic effects than the posterior pituitary snuff, containing 100 microgram AVP, and than 12.5 microgram synthetic LVP spray, but a shorter antidiuresis than 12.5 microgram vasopressin tannate in oil, administered intramuscularly, antidiuresis lasting 14, 6, 4 and 36 hs respectively. Chlorpropamide produced an inconstant and less potent antidiuresis. 10microgram DDAVP given per nostril twice a day cancelled completely and without side effects DI in five patients with bronchospastic reaction to-pituitary snuff; the same daily dose was sufficient for the safe treatment of two DI women along pregnancy and lactation periods. It is recommended to use DDAVP as elective drug for the treatment of cranial DI.