[The new diagnostic algorithms in early period of acute intoxications with psychotropic preparations].

The patients with acute intoxications with psychotropic preparations were analyzed for physical chemical characteristics of blood albumin at admission to hospital. The common level of reduced thiols in albumin fraction was analyzed too. The more frequent cases of unfavorable course of disease were noted at indicator rate less than 250 mkmol/l than at rates higher than 250 mkmol/l. This occurrence permits to consider this indicator as prognostically significant and to evaluate individual risk of unfavorable course of disease in the given category of patients. Under acute intoxications with psychotropic preparations appllying of fluorescent albumin test permits already at fist day of hospitalization to reveal a subgroup of patients with higher risk of subsequent development of pneumonia.

[Stress effect on the development of hemorrhagic stroke].

The objective of this work was to study how stress, activity in the open field test, and conformational properties of albumin-binding sites are associated with experimental hemorrhagic stroke in rats. The open-field behavioral pattern in rats was characterized by the previously developed by us activity index. In accordance with this activity index, rats were divided into two groups, i.e., active and passive animals. The animals were subjected to experimental hemorrhagic stroke with or without previous emotional stress. It was shown that the previous stress affected the stroke development. Stress loading before experimental stroke changed albumin conformational properties in rats with active and passive behavioral patterns in different ways. It was associated with different ability of the albumin globule to undergo pH-induced transition N-F and in different accessibility of albumin-bound fluorescent probe CAPIDAN to nitrate-induced fluorescence quenching.

[K-35, a fluorescent probe for albumin study: optical properties].

Fluorescent probe N-(carboxyphenyl)imide of 4-(dimethylamino)naphthalic acid, K-35, is used as an indicator of structural changes of human serum albumin molecules in pathology. The probe occupies albumin binding pockets where the probe environment is of very high polarity; probably, the pocket(s) contains protein polar groups and water molecules. At the same time rather small Stokes shift of K-35 fluorescence spectrum shows that the polar group motion is of one-two order of value lower than mobility of polar molecules in polar fluids. K-35 fluorescence decay in HSA can be described as a sum of three exponentials with time constants close to tau1=9 ns; tau2=3.6 ns and tau3=1.0 ns. A difference between excitation maxima of these three decay components shows that environment of these three species of K-35 molecules has been different before excitation. Different r values are probably a consequence of non-identical structure of several binding sites, or a binding site(s) can have a variable conformation.

[Features of the binding of the fluorescent probe K-35 to albumin].

The binding of the fluorescent probe K-35 (CAPIDAB, N-(carboxyphenyl)imide of 4-(dimethylamino)naphthalic acid), which is used as an indicator of albumin structural changes in pathology, to human serum albumin has been studied. Based on the data on the fluorescence decay of the probe, four types of sites of binding of K-35 to albumin have been recoonized, which differ by fluorescence decay time (tau) and binding constants (K). Probe molecules bound to the first type of sites have a decay time close to 8-10 ns; this value corresponds to a high fluorescence quantum yield of about 0.7. These sites have a maximal binding constant, K1 = 5 x 10(4) M(-1). Tau2 of the second type of sites is close to 3.6 ns and K2 = 1 x 10(4) M(-1), which is much lower than K1; however, the number of the sites is several times greater. The number of sites of the third type and the binding constant are close to those of the second type, but the decay time tau3 is equal to 1 ns, which is significantly lower than tau2. The binding of K-35 to sites of the second and the third types is characterized by a positive cooperativity. Their properties are similar but not completely identical. The total number of sites of these three types is about 2 per one HSA molecule. There are one to two sites of the fourth type where bound K-35 molecules have a very low decay time tau4 << 1; i.e. they are virtually nonfluorescent, and K4 = 1 x 10(4) M(-1). The major contribution to the steady-state fluorescence is made by probe molecules bound to sites of the first and second types. As a rule, the concentration of albumin binding sites in blood is significantly higher than the concentration of metabolites and xenobiotics transferred by albumin. Therefore, this metabolite or the probe in these experiments, is distributed between different sites in accordance with their K(i)n(i) values (n(i) is the number of sites of the ith type per albumin molecule). It was shown that the low occupation of the sites leads to an approximately equal number of K-35 molecules bound to different sites of types 1, 2, and 3. The competition of K-35 with phenylbutazone, a marker of the albumin drug-binding site I, allows one to suggest that the K-35 site of the first type is localized near the drug site I, while the sites of the second and third types are close to it.

[Disturbance of monoaminooxidase activity and indices of endogenous intoxication in patients with the first episode of schizophrenia].

An aim of the paper was to study some biochemical parameters of drug-naïve patients with the first episode of schizophrenia. Activities of platelet monoaminooxidase (MAO) and semicarbazide, a sensitive blood serum aminooxidase (BSA), levels of middle-sized molecules (MSM) and malonic dialdehyde (MDA), parameters of functional state of serum albumin were assessed in 16 patients. Severity of symptoms in patients with the first episode of schizophrenia was assessed as moderate (PANSS scores 73.1+/-12.5) before the treatment. The increase of MAO by 107%, reduction of BSA by 29% and increase of MSM level by 140% was found in patients compared to controls (p<0.01). The study of other biochemical (MDA level) and biophysical (effective albumin concentration) parameters did not yield unequivocal results. It has been suggested that MAO and BSA are integral components of pathogenetic mechanisms in patients with the first episode of schizophrenia.

[Conformational disturbances of serum albumin binding sites in schizophrenia].

The properties of serum albumin binding sites were studied using quenching of fluorescence of the molecular probe CAPIDAN (N-carboxyphenylimide of dimethylaminonaphthalic acid) with the nitrate anion. The samples of serum were obtained from 24 patients with paranoid schizophrenia and 24 healthy volunteers. In the absence of quencher the specific probe fluorescence was 1,4 times higher in patients than in volunteers. Fluorescence quenching constant for the probe bound to albumin was (M+/-m) 2,48+/-0,17 l/mol in patients versus 4,65+/-0,37 l/mol (p<0,01) in volunteers (p<0,01). The fluorescence fraction assessable to quenching was significantly (p<0,01) lower in schizophrenic patients as compared to controls (0,60+/-0,03 and 0,76+/-0,03) respectively). Thus, it is shown that in patients with schizophrenia the conformational state of albumin binding sites is significantly changed as compared to controls that can lead to the changes in the protein-ligand interaction and, thus, contribute to the pathogenesis of the disease and patient's response to treatment.

[A study of ligand binding to protein using resonance energy transfer from the isoindole fluorescent label to ligand]. / Issledovanie sviazyvaniia ligandov s belkom s pomoshch'iu perenosa énergii s izoindol'noi fluorestsentnoi metki na ligand.

A method for studing the binging of ligands absorbing the light in the region of 350-550 nm to protein is described. The method is based on resonance energy transfer between the fluorescent label covalently bound to the protein and the ligand. The isoindole label, a product of the reaction of the protein with o-phthalaldehyde in the presence of 2-mercaptoethanol, was used as a fluorescent donor. The method was used to determine the binding parameters of a fluorescent probe (a naphthalimide derivative) with human serum albumin. A comparison of the results obtained by the resonance energy and transfer by equilibrium dialysis showed a high accuracy of the resonance energy transfer method.

[The prognostic value of albumin fluorescence test in the evaluation of outcome of acute intoxication with psychotropics]. / Prognosticheskaia tsennost' al'buminovogo fliuorestsentnogo testa v otsenke iskhoda ostrykh otravlenii psikhotropnymi sredstvami.

Two hundred and five patients with toxicogenic and somatogenic intoxication with psychotropics (PT) were examined for the purpose of evaluating the prognostic value of the albumin fluorescence test. An analysis of distributions of laboratory indices in the groups of survived (n = 176) and diseased (n = 29) patients demonstrated the following: the parameter "efficient albumin concentration" (EAC) measured by fluorescence is informative in respect to prognosticating an outcome of PT intoxication of both toxicogenic and somatogenic forms. EAC provides for isolating a subgroup of patients with the risk of unfavorable outcome that is 3-4-fold higher versus the mean value. The routine laboratory parameters, e.g. level of creatinine, serum urea, are rather of a less prognostic significance versus the EAC fluorescence parameter.

[Informative value of the parameter "effective concentration of albumin" in diffuse peritonitis: data from a multi-center study]. / Informativnost' pokazatelia "éffektivnaia kontsentratsiia al'bumina" pri rasprostranennom peritonite: dannye mnogotsentrovogo issledovaniia.

The paper represents data on new potentialities in evaluating a severity of patients with total peritonitis (TP). They are based on the fluorescence analysis of the physical-and-chemical properties of blood albumin. The findings are a result of a multi-center research undertaken by clinics of Moscow and Moscow Region. The properties of albumin-molecule binding sites were investigated by the fluorescence test. The parameter, measured in the test, was named "effective albumin concentration". The study protocol comprised the data of clinical examinations, scales SAPS and MPI as well as laboratory findings and EAC. The subsequent analysis denoted that a combination of 2 parameters, i.e. EAC and serum urea concentration routinely measured, is highly informative in evaluating the TP severity and correlates well with the survival rate in the discussed pathology. The study results could be a basis for an early individual prognostication of TP course made on the 1st postoperative day.

[Diagnostic significance of albumin transporting characteristics and blood level of troponin T in myocardial infarction]. / Diagnosticheskoe znachenie transportnykh svoistv al'bumina i soderzhaniia v krovi troponina T pri infarkte miokarda.

Serum albumin binding capacity (ABC) was studied using K-35 hydrophobic fluorescent probe in order evaluate the passive transfer and absorption of saturated fatty acids (s-FA) by the cells. ABC decreases on day 3 of myocardial infarction and remains low till discharge and is paralleled by hyperTG. We believe that ABC decrease reflects activation of passive transfer and absorption of s-FA by the cells during simultaneous blocking (presumably by acute phase proteins) of active apoE/B-100 receptor endocytosis of s-FA in the form of TG as VLDL remnants. Decrease of ABC reflects a later than erythrocyte sedimentation rate and acute phase proteins normalization of metabolic consequences of inflammation. ABC helps indirectly evaluate the blocking of active (receptor) absorption of s-FA in the form of TG and activation of passive absorption of s-FA from associations with albumin by the cells.

[Size of a human serum albumin molecule in solution]. / Razmer molekuly syvorotochnogo al'bumina cheloveka v rastvore.

The size of a human serum albumin molecule in aqueous solution containing 150 mM NaCl was studied using small-angle neutron scattering. The molecular radius of gyration was estimated to be 27.4 +/- 0.35 A. The compact sphere should have a smaller radius of gyration, whereas the popular human serum albumin model, a "cigar" 136 A long, should correspond to a greater radius of gyration. Possible shapes of the human serum albumin molecule which are in accordance with the results obtained, are the following: an extended ellipsoid less than 110 A of length or a nonsymmetrical oblate ellipsoid with a diameter of 85 A. The oblate ellipsoid might be close to the heart"-shaped structure of the crystalline human serum albumin molecule. The size of the albumin molecule does not change significantly as pH increases to 8.9. The possibility of the dynamic coexistence of various human serum albumin conformers in solution is discussed.

[Method for assessing involution ophthalmoendotoxicosis by fluorescent analysis of lacrimal fluid]. / Sposob otsenki involiutsionnogo oftal'moéndotoksikoza putem fliuorestsentnogo issledovaniia sleznoi zhidkosti.

A method for evaluating the binding capacity of aqueous humor albumin as endoophthalmotoxicosis indicator is proposed, consisting in evaluation of this parameter in the lacrimal fluid by fluorescent probe K-35. A correlation between the studied parameter in aqueous humor, lacrimal fluid, and blood serum has been detected. Albumin binding capacity in the above biological fluids of patients with involution ophthalmopathy was low.

[Activity of the autoimmune process during systemic rheumatic diseases and distribution of constant brain potential]. / Aktivnost' autoimmunnogo protsessa pri sistemnykh revmaticheskikh zabolevaniiakh i raspredelenie postoiannogo potentsiala golovnogo mozga.

The data obtained upon examination of patients with systemic rheumatic diseases were analyzed by the methods of multiple regression and step-by-step discriminant analysis. Is was shown that the activity of autoimmune process correlates with the parameters of distribution of constant potential level in the brain of each patient, irrespective of diagnosis. The scatter of potential values was the most important predictor of activity. An increase in the activity of the autoimmune process correlated with a decrease in the scatter of potential values at recording points; the cerebral cortex became equipotential.

[Constant brain potentials as neurophysiological markers of autoimmune process]. / Postoiannye potentsialy golovnogo mozga kak neirofiziologicheskie markery autoimmunnogo protsessa.

Neuroimmune interactions in systemic rheumatic diseases were studied. The state of the central nervous system was assessed from the parameters of constant brain potentials, and the state of the immune system, from a complex of immunobiochemical parameters. The highest multiple correlation coefficients were revealed between the immunobiochemical parameters and the parameters of the constant brain potential, which characterize linear and standard deviations of potentials in temporal zones from potentials at other points of recording. The results are discussed in terms of structural and functional integration of the immune and nervous systems.

[Albumin binding capacity and blood lipids during the development of acute myocardial infarct]. / Sviazyvaiushchaia sposobnost' al'bumina i lipidy syvorotki krovi pri razvitii ostrogo infarkta miokarda.

Excellent reproducibility of rapid fluorescent methods for assessment of cholesterol and triglycerides, effective and total concentrations of albumin permits monitoring these parameters over the course of development of acute myocardial infarction. Manifest changes of cholesterol and triglycerides and of albumin binding reserve estimated as the ratio of effective to total albumin concentrations multiplied by 100% were observed in acute myocardial infarction and even in a single anginal attack. Total albumin concentration changed on days 3-4 only in infarction. Time course of the parameters was related to the disease severity.

[Fluorescent method of determining mass concentration of human serum albumin]. / Fliuorestsentnyi sposob opredeleniia massovoi kontsentratsii al'bumina syvorotki krovi cheloveka.

Fluorescent method for measuring mass (total) albumin concentration in human blood serum is suggested. Fluorophore K-35 previously suggested for measuring the effective concentration of albumin is recommended. Total albumin concentration is measured in acid pH range in the presence of nonionic detergent. Conditions under which the effects of factors impeding albumin assay virtually do not manifest were found. The resultant values of total albumin concentration coincide with the values determined by the bromocresol purple method.

[Effects of haloperidol therapy on dynamics of serum albumin binding capacity in patients with schizophrenia]. / Vliianie terapii galoperidolom na dinamiku pokazatelei sviazyvaiushchei sposobnosti al'bumina syvorotki krovi u bol'nykh shizofreniei.

Time course of blood serum albumin binding capacity was studied by the fluorescent method in schizophrenics, including those treated with haloperidol. Effective and total albumin concentrations were measured and binding reserve estimated, which was equal to ratio of effective to total albumin concentrations multiplied by 100%. Effective albumin concentrations and albumin binding reserve were found markedly reduced in the patients vs. healthy donors. Drastic changes of albumin binding reserve from one examination to another and a trend to increase of effective concentration of albumin were characteristic of patients administered haloperidol. In the patients administered no haloperidol albumin binding reserve was as a rule unchanged and effective albumin concentration had a trend to reduction.

[Characteristics of the method and reagents in fluorescent determination of effective and total concentration of serum albumin]. / Nekotorye kharakteristiki metoda i reaktivov dlia fliuorestsentnogo opredeleniia éffektivnoi i obshchei kontsentratsii syvorotochnogo al'bumina.

The characteristics of fluorescent essay of total and effective albumin concentrations by Zond-Albumin kit (experimental lot) were as follows: intraseries reproducibility in measurements not more than 1.6%, the error in serum dosage being mainly responsible for the total error. After unsealing the ampules the reagents may be stored at 5 degrees C for at least a week. Fluorescence intensity recorded in measurements in the majority of samples is stable for at least 50 min after addition of reagents. Measured values of total albumin concentration coincide with the results of measuring mass albumin concentration by bromocresol blue method, coefficient of linear correlation r = 0.98.