[Immunoassay of nine serological tumor markers on hydrogel-based microchip].

A prototype of test-system for simultaneous quantitative assay of nine tumor markers in blood serum was developed. The main constituent of the test-system is OM-9 biochip containing immobilized antibodies against nine oncomarkers: α-fetoprotein (AFP), carcinoembryonic antigen (CEA), human chorionic gonadotropin (HCG), cancer antigen 15-3 (CA 15-3), cancer antigen 125 (CA 125), cancer antigen 19-9 (CA 19-9), prostate-specific antigen, total (PSAtot) and free (PSAfree) forms, neuron-specific enolase (NSE). The biochip-based assay procedure for carrying out simultaneous quantitative determination of nine tumor markers in patient's blood serum: two-steps sandwich-immunoassay, was proposed. The main analytical characteristics of the method were obtained. The results permit to consider the prototype of the test-system as a promising instrument for clinical application. The test-system prototype was tested using blood serum samples of oncological patients (252 samples) and healthy donors (185 samples). Increased concentrations of one or more tumor markers above the normal level were found in 76.6% cases of oncological patients and only in 6% cases of healthy donors. For colorectal cancer patients group, application of modern statistical methods of data-processing in medical researchers, i.e. ROC-analysis and logistic regression, indicted that the simultaneous assay of nine tumor markers on biochips showed much more diagnostic significance (area under the ROC-curve (AUC) reached 0.84) than traditional assay of 2 tumor markers, CEA and CA 19-9 (AUC = 0.59). The developed biochip-based test-system can be recommended both for the estimation of people's health, e.g., for standard medical examination, and for tracking of tumoral process in postsurgical period or after specific tumor treatment.

[Assessment of rate of infection with agents of bacterial infections in ticks captured on one of the Moscow park terrains].

AIM: To study the rate of infection of ticks captured one of the Moscow park terrains with bacteria (agents of tick borreliosis and anaplasmosis). MATERIALS AND METHODS: Rates of infection of dried ticks with agents of main tick-borne bacterial infections (tick borreliosis and anaplasmosis) were determined by nested PCR. RESULTS: In May-June 2006, 76 ticks (40 adult females, 36 adult, males) belonged to Ixodes ricinus species were captured by the method "on flag". Number of ticks on the chosen terrain was 1.77 ticks per km2. 22.4% of ticks (12 females and 5 males) were positive for the agent of tick borreliosis--spirochete Borrelia burgdorferi sensu lato which is pathogenic for humans. The main detected pathogen was Euro-Asian genovariant of B. garinii--7 female and 5 male ticks (70.6% from total number of infected ticks) were infected with it. Five female ticks were infected with genovariant of B. afzelii. One female tick (1.2%) was infected with B. valaisiana. CONCLUSION: Anaplasma A. phagocytophilum causing human granulocytic anaplasmosis was not detected in captured adult ticks.

[Development of quantitative immunoenzyme and radioimmunologic analysis of lambda-free immunoglobulin light chains]. / Razrabotka kolichestvennogo immunofermentnogo i radioimmunologicheskogo analiza lambda-svobodnykh legkikh tsepei immunoglobulinov.

New quantitative techniques of radioimmunoassay (RIA) and enzyme immunoassay (EIA) were developed for determination of free light lambda-chains of immunoglobulins (immunometric sandwich-like variants) in biological fluids using two types of monoclonal specific antibodies (MAB-1 and MAB-2) to different epitopes of the antigen molecule: MAB-1 were immobilized on a solid phase (polystyrene beads), whereas MAB-2 were labeled with iodine or with the enzyme. The test systems prepared can be used for determination of concentrations from 25 to 1000 ng/ml, are very sensitive (25 ng/ml), and the analysis time is 5 h. The two methods were compared, and their clinical and diagnostic validity was evaluated in patients with various diseases associated with disorders in the antibody synthesis by the immune system.

[Antigliadin antibodies in the absence of celiac disease]. / Antigliadinovye antitela v otsutstvie tseliakii.

Antigliadin antibodies (AGA) mark celiac disease, but AGA are also encountered in IgA-nephritis, psoriasis, sickle-cell anemia, hepatic disorders, juvenile rheumatoid arthritis, autoimmune thyroidism and in persons who occupationally contact great amounts of wheat. AGA IgA and/or IgG were registered in 19 of 60 subjects (51 adults and 9 children) with various immunomediated diseases without symptoms of celiac disease: in 4 cases of chronic active hepatitis, in 2 of 4 cases of chronic persistent hepatitis, in 4 of 16 cases of rheumatoid arthritis, in 3 of 19 cases of IgA-deficiency, in 1 of 8 cases of SLE, in 2 cases of postvaccine reaction, in all the single cases of juvenile rheumatoid arthritis, focal scleroderma, macroglobulinemia. IgA only occurred in in 6 patients, IgG- in 6 patients, both IgA and IgG in 7 patients. The most pronounced positive reaction to AGA was recorded in 8-year-old girl with juvenile rheumatoid arthritis. The emergence of AGA in immunomediated diseases may be attributed to the response to food protein in pathological conditions and is often unrelated closely with celiac disease.

[The evaluation of the ratio of free and total serum prostate-specific antigen as an additional method in the diagnosis of prostatic cancer]. / Otsenka sootnosheniia svobodnogo i obshchego syvorotochnogo prostaticheskogo spetsificheskogo antigena kak dopolnitel'nogo metoda v diagnostike raka prostaty.

The conventional method of prostatic cancer monitoring by determination of serum prostatic specific antigen (PSA) was compared to the method of free-to-total PSA correlation. The latter proved more informative concerning prostatic cancer.

[Monoclonal free light-chain immunoglobulins: their clinical interpretation]. / Monoklonal'nye svobodnye legkie tsepi immunoglobulinov: klinicheskaia interpretatsiia.

23 cases of free light chains (FLC) occurrence in the blood and/or urine in the absence of monoclonal gammopathy are reported. Of them 15 cases were attributed to renal diseases associated with deranged catabolism of low-molecular proteins including FLC. In the rest cases FLC hyperproduction in autoimmune diseases was suspected to cause FLC appearance. No correlation was found between the amounts of FLC, creatinine, beta 2-microglobulin, light chains, kappa/lambda proportions.

[Changes in the levels of serum proteins in patients with chronic obstructive bronchitis]. / Otsenka izmenenii urovnei belkov syvorotki krovi u bol'nykh khronicheskim obstruktivnym bronkhitom.

A new approach is suggested for evaluation of changes observed in serum levels of immunoglobulins, compliment components and proteinase inhibitors in subjects suffering from chronic obstructive bronchitis (COB). These levels were compared for COB patients (n-15) in remission and under aggravation of the disease. The comparison promoted a significant assessment of the shifts and their pattern for humoral defence factors. There appeared a correlation between concentration of some serum proteins and bronchial obstruction characteristics.

[A possible variant of a false diagnosis of infection with the human immunodeficiency virus]. / Vozmozhnyi variant lozhnoi diagnostiki infitsirovaniia virusom immunodefitsita cheloveka.

The authors describe a patient who demonstrated positive blood responsiveness to the nuclear antigens of human immunodeficiency virus (HIV) (p17, p31 and p55), observed steadily for 1 year and 4 months. The donor's disease history consideration made it impossible to include him in one of the known groups at risk for HIV infection whereas the lack of any changes in immunoblotting enabled one to exclude the diagnosis of HIV infection. The given case and other similar cases form the basis for introducing the second parallel screening during blood testing for HIV infection to bar the use of such blood for transfusion.

[Atopic dermatitis. IV. The role of proteinase inhibitors in the pathogenesis, assessment of the severity and the prognosis of the disease]. / Atopicheskii dermatit. IV. Rol' ingibitorov proteinaz v patogeneze, otsenke tiazhesti i prognoze zabolevaniia.

Examinations of 59 patients with atopic dermatitis have revealed increased content of the major inhibitors of proteinases, a relationship between the patients' age and alpha 2-macroglobulin level in both the patients and the reference group normal subjects. Detection of correlations between alpha 1-proteinase inhibitor and serum IgE as well as between C1 inactivator and C3c have lead to a supposition on the contribution of proteinase inhibitors in the modulation of immune reactions. Clinical features of atopic dermatitis in alpha 1-proteinase inhibitor deficiency and in various blood serum IgE levels are discussed.

[Atopic dermatitis. I. Characteristics of the clinical course and state of the immune status in relation to the initial level of serum IgE]. / Atopicheskii dermatit. I. Osobennosti klinicheskogo techeniia i sostoianiia immunnogo statusa v zavisimosti ot iskhodnogo urovnia IgE syvorotki.

Altogether 97 patients with atopic dermatitis have been examined to follow this relationship, 72 patients presented with disseminated efflorescence, 25 patients with local rash. Measurements of immunoglobulins A, M, G, and D, of the C3c complement components, of the total and allergen-specific IgE C4, inactivators have been carried out, as well as of the total lymphocyte, B lymphocyte, and CD3+, CD4+, and CD8+ lymphocyte subpopulations; the lymphocyte response to ConA mitogenic stimulation has been also under study. The patients with normal and elevated levels of IgE differed by the clinical picture of the disease and by case histories, as well as by the immunity parameters and by the results of zaditen therapy. Cases with very low IgE levels may be indicative of alternative pathogenetic mechanisms.

[Atopic dermatitis. II. The status of complement proteins and the pathogenetic role of anaphylatoxins C4a, C3a and C5a]. / Atopicheskii dermatit. II. Sostoianie komplementarnykh belkov i patogeneticheskaia rol' anafilatoksinov C4a, C3a i C5a.

Eighty-eight patients with atopic dermatitis have been examined for the complementary proteins C3 (C3c), C3act., C4, C1inact. by radial immunodiffusion; in 36 of these patients anaphylatoxins C3a des Arg, C5a des Arg, and C4a des Arg have been radioimmunoassayed. Increased levels of C4a des Arg have been revealed in 50% of the examinees. C3a des Arg levels did not differ significantly from the reference values, and C5a des Arg level has been within the normal range in all the patients. C4a des Arg has been related to the severity of the skin inflammation and did not depend on the serum IgE content. C3 (C3c), C3act., C1inact. have been elevated and C4 reduced. These findings evidence activation of the complementary system in atopic dermatitis and indicate the role of anaphylatoxin C4a des Arg in the maintenance of the inflammatory reactions.

[Place of carbohydrate antigens in the spectrum of protein tumor markers in patients with tumors at different sites]. / Mesto karbogidratnykh antigenov v spektre belkovykh onkomarkerov u bol'nykh s opukholiami razlichnykh lokalizatsii.

Carbohydrate antigens (CA-19.9 and CA-125) levels were assayed in patients suffering rectal, ovarian, breast and hepatic carcinoma, severe recurrent hepatitis A, chronic persistent hepatitis B, hepatic cirrhosis and in cases of normal pregnancy. Patients with early-stage cancer usually failed to reveal the said antigens. This was also the case in some patients with clinically manifest tumor. Clinical application of said assay may improve postoperative monitoring cancer patients. Cancer was more frequently associated with increased level of CA-19.9 than with that of CA-125.

[Dynamics of hormones, sugar and electrolytes during hypodynamia according to blood biochemical indices]. / Dinamika gormonov, sakhara, élektrolitov v usloviiakh gipodinamii po dannym biokhimicheskikh pokazatelei krovi.

Time-course variations in hormones, carbohydrates and electrolytes (specifically, cortisol, aldosterone, testosterone, T3, T4, sugar, potassium, sodium and chlorides) in blood of essentially healthy men, aged 19-59 years, kept for 30 days in a closed life-support system were measured. The hormones, sugar and electrolytes varied in a different manner. The subjects, aged 48-59 years, who performed regular exercises showed a higher stability of potassium, sodium and chlorides and a normalization of hormones by test day 30 in contrast to other groups of subjects.