[Experimental data on avian influenza virus detection using magnetic immunosorbents].

To increase the sensitivity and specificity of detecting high-pathogenic avian influenza variant (HSN1), laboratory studies were conducted at low virus concentrations in water samples, by using magnetic immunosorbent (MIS) test systems and selective avian influenza virus concentrating units. MIS-based selective virus concentrating, followed by rapid assays (ELIZA and RT-PCR), detect the low concentrations (as low as 10 nm in 5,000 ml of water) of avian influenza A/H5N1 antigen and RNA. The method developed opens new avenues for indication of an avian influenza pathogen in different environmental objects, including the water of surface water reservoirs of unlimited volume, with varying pollutions and low virus concentrations.

Characterization of urinary peptide biomarkers of acute rejection in renal allografts.

We previously demonstrated that 4.7 kDa and 4.4 kDa peptides are useful in diagnosing acute rejection in renal transplant recipients. The aim of this study was to characterize these polypeptides and assess their potential as biomarkers. The polypeptides were identified as human beta-Defensin-1 (4.7 kDa) and alpha-1-antichymotrypsin (4.4 kDa), by tandem mass spectrometry and ProteinChip immunoassay. The urinary abundance of both polypeptides, assessed using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS), revealed a reduction in beta-Defensin-1 while alpha-1-antichymotrypsin increased in patients with rejection (p < 0.05) compared with clinically stable transplants. The area under the curve (AUC) for the receiver operator characteristic (ROC) curve for the diagnosis of rejection for the ratio of both peptides combined was 0.912. Longitudinal analysis confirmed a reduction in beta-Defensin-1 with a reciprocal increase in alpha-1-antichymotrypsin as rejection developed. The difference in urinary beta-Defensin-1 levels quantified by radioimmunoassay was 176.8 +/- 122.3 pg/mL in stable patients compared with 83.2 +/- 52.2 pg/mL in patients with acute rejection, with an ROC AUC of 0.749 (p < 0.01). Immunohistochemistry (IHC) confirmed reduced beta-Defensin-1 expression in the renal parenchyma of patients experiencing acute rejection. In conclusion, the ratio of beta-Defensin-1 and alpha-1-antichymotrypsin excretion in the urine is a novel, potentially useful candidate biomarkers of acute rejection.

[Toxic effect of the novel phenol antioxidant SO-3 in the subchronic experiment]. / Izuchenie toksicheskogo deistviia novogo fenol'nogo antioksidanta CO-3 v subkhronicheskom éksperimente.

Subchronic tests on rats showed that a new phenolic antioxidant bis[3,(3, 5-di-tert-butyl-4-hydroxyphenyl)propyl] sulfide (SO-3) exhibits low toxicity upon peroral administration. Neither significant variations in the hematological and integral indices, nor pathological shifts in the activity of serum enzymes or pathomorphological changes in the internal organs were observed. Simultaneous administration of a sunflower oil with SO-3 led to a significant decrease in the blood cholesterol level, produced a hepatoprotector effect, and exhibited a protective action with respect to the mucous membranes of the gastrointestinal tract. Animals receiving large doses of SO-3 showed no signs of a prelipid stage of the arteriosclerosis typically observed in the control group.

5-[3-(E)-(4-azido-2,3,5,6-tetrafluorobenzamido)propenyl-1]-2'-deoxy- uridine-5'-triphosphate substitutes for thymidine-5'-triphosphate in the polymerase chain reaction.

The DNA targets may be labeled and simultaneously amplified in the polymerase chain reaction (PCR) using a pair of respective primers after elongation with nucleoside-5'-triphosphates carrying photoreactive groups. The amplified DNA may be subsequently photoactivated by irradiation above 300 nm, resulting in photo-cross-linking of the strands. For this goal 5-[3-(E)-(4-azido-2,3,5,6-tetrafluorobenzamido)propenyl-1]-, 5-{N-[N'-(4-azido-2,3,5, 6-tetrafluorobenzoyl)-3-aminopropionyl]aminomethyl}-, and 5-{N-[N'-(2-nitro-5-azidobenzoyl)-3-aminopropionyl]aminomethyl}-2'-de oxyuridine-5'-triphosphate (VII, VIa, and VIb) derivatives have been synthesized. It was found that VII is capable of efficiently elongating DNA primers with both Klenow fragment DNA polymerase I and Thermus aquaticus DNA polymerase. Thereto, it turned out to provide quantitative incorporation in DNA as revealed by the formation of the full-length amplificate by PCR in the presence of this photoreactive analogue without any dilution with natural dTTP. On the contrary, it was found, that incorporation of VIa and VIb do not permit further DNA replication.

[Highly-sensitive nonradioactive detection of the tick-borne encephalitis virus]. / Vysokochuvstvitel'naia neradioaktivnaia detektsiia virusa kleshchevogo éntsefalita.

The non-radioactive reverse dot-blot method was used for the detection of tick-borne encephalitis virus (TBEV) in clinical specimens. The method involves reverse transcription (RT) and polymerase chain reaction (PCR) using a pair of biotin-labelled oligonucleotide primers. These primers flank a region in the gene of the envelope protein E, which is more conserved than other regions, and initiate the polymerisation with RNAs of all the investigated strains. The amplified cDNA was captured from solution on a solid support using complementary oligonucleotides covalently bound to a polyamide membrane. The biotin labels of the resulting hybrids were visualized by means of the streptavidin-horseradish peroxidase conjugate. The detection limit of the test was about 10(3)-10(4) molecules of target RNA. The sensitivity was comparable to that obtained by dot-hybridization of PCR-product with 32P-labelled DNA probe. The method was used for the detection of RNA in specimens of tick and blood.