[Affine magnetic sorbents supported on coal ash microspheres for recombinant protein isolation].

The results of the development and utilization of an affine magnetic sorbent with Ni2+ ions immobilized on coal ash microspheres are reported. The applicability of the material in the isolation of Histag proteins is demonstrated by examples of the recombinant green fluorescent protein from Clytia gregarium and the Ca2+ regulated photoprotein obelin from Obelia longissima. The specific sorption capacity of the sorbent was 2-7 mg/cm3 for medium-size proteins (20-30 kDa). The particles are suitable for chromatography with the presence of chaotropic agents and EDTA. They are easy to manipulate as isolation of a target protein takes 30-35 min. On one hand, the elevated affinity of the sorbent to proteins rich in native histidines may result in a high degree of irreversible sorption; on the other hand, it allows isolation of such proteins without the introduction of artificial polyhistidine tracts.

[A highly sensitive and rapid method for the detection of DNA fragments using the photoprotein obelin as a reporter].

The recombinant Ca2+-activated photoprotein obelin was used as a reporter protein in a solid-phase bioluminescent hybridization DNA assay. Oligonucleotide probes were immobilized on the surface of a fine-grained polymer or microbiological plates of different types. A 30-mer oligonucleotide or its derivative with a biotin residue on the 3'-terminus, as well as a denatured double-stranded PCR fragment of the hepatitis C virus with the sequence of the 30-mer oligonucleotide, was used as a DNA template. The probe in the hybridization complex was labeled by elongation of the chain using Taq DNA polymerase in the presence of biotinylated deoxyuridine triphosphate. The results of the bioluminescent assay were compared with the results of colorimetric analysis obtained with alkaline phosphatase as a reporter protein. It was shown that the use of the bioluminescent obelin label substantially accelerates the DNA detection procedure, ensures a high sensitivity of the assay (no less than 10(-15) mol of DNA template), and enables quantitative determination of the amount of DNA template in the tested sample.

[The unique course of a chronic generalized infection with the hepatitis B virus (a clinico-morphological observation)]. / Svoeobraznoe techenie khronicheskoi generalizovannoi infektsii virusa gepatita B (kliniko-morfologicheskoe obsledovanie).

In a young male chronic generalized HBV infection (acute viral hepatitis in childhood, HBsAg, HBeAb, HBcAb in the blood serum) ran with a long-term fever and involvement of many organs and systems (the liver, lungs, CNS) complicating the diagnosis. The patient died in the presence of CNS affection and hepatorenal insufficiency. At biopsy and autopsy it was established that the patient had active hepatic cirrhosis, fibrosing alveolitis, pulmonary vasculitides, chronic pneumonia, cerebral vasculitis, myocarditis and postmyocarditis cardiosclerosis, necrotizing myositis, Sjogren's syndrome, mesangioproliferative glomerulonephritis.

[Antibodies to the structural proteins of the measles virus studied in some chronic diseases]. / Issledovanie antitel k strukturnym belkam virusa kori pri nekotorykh khronicheskikh zabolevaniiakh.

The qualitative and quantitative analysis of antibodies to measles virus (MV) structural proteins (SP) in sera from patients with chronic glomerulonephritis (CGN), chronic active hepatitis (CAH), and liver cirrhosis (LC) was done. The patients were shown to have neutralizing antibody titres (NAT) higher than those in healthy subjects. An analysis of antibodies to SP was carried out by the radioimmunoprecipitation assay. Antibodies were detected to hemagglutinin, nucleoprotein (NP), fusion protein and to matrix protein (M) both in sera from the patients with these chronic diseases, healthy subjects, and patients with active measles. (The two latter groups were selected for comparison). However, some patients with CAH and LC had no antibodies to M protein in spite of very high NAT. The quantitative analysis of MV antibodies to SP was done only for NP because this antibody had the least individual variations. The quantity of anti-NP antibodies was higher in most sera from patients with chronic diseases than in those from healthy subjects, and reached the level of that in patients with active measles. The presence of MV genome in the peripheral blood lymphocytes from patients CAH, CGN, and LC had been shown earlier. So it is assumed that MV persists in lymphoid tissue where the expression of all SP genes is realized.

[Expert research on sera yielding false-positive results for HIV antibodies during screening]. / Ekspertnye issledovaniia syvorotok, daiushchikh lozhnopolozhitel'nye rezul'taty na antitela k VICh pri skrininge.

The significance of different serological methods and assay systems for the verification of false positive cases of HIV infection has been analyzed on the basis of materials obtained in arbitration studies. As demonstrated by this analysis, the use of such highly specific and sensitive systems as Huma-Lab, Enzygnost, Serodia and Erythrorecombinant has made it possible to obtain a reliable result as early as at the first stage of expert diagnosis in the enzyme immunoassay and the agglutination test. The methods of radioimmunoprecipitation and indirect immunofluorescence have permitted a more precise differentiation of doubtful results than that achieved by immune blotting.

[Effect of late consequences of radiation on shortening the life-span]. / Vliianie otdalennykh posledstvii oblucheniia na sokrashchenie prodolshitel'nosti zhizni.

Experiments were conducted on mongrel male albino rats injected with 90Sr (12.9 kBq/g) and 45Ca (48.0 kBq/kg). It was shown that death of animals from lung inflammations markedly influenced the average life of a cohort.