[Wide range of the use of natural lipases and esterases to inhibit Mycobacterium tuberculosis]. / O vozmozhnostiakh primeneniia prirodnykh lipaz i ésteraz dlia ingibirovaniia Mycobacterium tuberculosis.

Lipases and/or esterases (hereinafter referred to as esterases) isolated from the wax moth (Galleria mellonella) were found to have a bacteriological action on Mycobacterium tuberculosis (MBT) H37Rv. Different types of raw esterase preparations (REP) were incubated with MBT at 37 degrees C for 18 hours, the incubate was seeded on the Finn-II solid medium or intraperitoneally injected into guinea pigs in a single dose of 100,000,000 bacteria. There was no growth of MBT in the medium within 8 weeks, some variants of REP causing a destruction of the medium for 3-7 days. This "toxic" effect on the lipid-containing Finn-II medium could be lowered by the simplest techniques for purifying esterases. In the experimental guinea pigs, a tuberculous process substantially regressed: autopsy of control animals at interval of 3-6 weeks after inoculation with native MBT showed the typical picture of progressive generalized tuberculosis; at the same time a visible pathology was not noted in the animals contaminated with MBT incubated with esterases. At week 7, control guinea pigs died; the onset of a tuberculous process was observed in experimental guinea pigs at week 8. An attempt to reveal the therapeutical effect of REP on guinea pigs with tuberculosis in a direct experiment failed. At the same time, there was a low toxicity of REP (in the used doses) for guinea pigs and B10.A(4R) mice. Based on their own findings and some data available in the literature, the authors have arrived at the following provisory conclusions: the studied REPs contain mammalian lysosomal lipase-type enzymes that determine the bacteriostatic and, perhaps, bacteriolytic effect of REP shown on MBT in vitro; there is evidence for promises of continued detailed studies of natural esterases for searching new antituberculous agents. A program of investigations of the studied and other natural esterases has been developed by taking into account the authors' developments and know-how. The study may be regarded as part of global unselected screening of biological and other materials for detecting new promising sources of drugs.

[Oligomerization of phenylalanine hydroxylase during its activation with phenylalanine]. / Oligomerizatsiia fenilalaningidroksilazy pri ee aktivatsii fenilalaninom.

Activated and nonactivated forms of phenylalanine hydroxylase varied in the activity within the first minutes of the reaction initiated by means of a synthetic coenzyme 6,7-dimethyl-5,6,7,8-tetrahydropteridine. In activation of the enzyme by phenylalanine the degree of its oligomerization was altered. As shown by chromatography and ultracentrifugation nonactivated enzyme was a dimer with molecular mass 130,000 daltons, whereas the activated form was found to be tetramer of 260,000 daltons. H and L subunits of phenylalanine hydroxylase were equally important in organization of the tetramer. Activation of phenylalanine hydroxylase altered the enzyme affinity to hydroxyapatite. Thus, allosteric activation of phenylalanine hydroxylase occurred due to the enzyme oligomerization.

[Active form of dihydropteridine reductase in human chorion cells. Possibility of prenatal diagnosis]. / Aktivnaia forma digidropteridinreduktazy v kletkakh khoriona cheloveka. Vozmozhnost' prenatal'noi diagnostiki.

Activity of dihydropteridine reductase was studied in human chorion cells for development of accurate methods for prenatal diagnosis of phenylketonuria (especially its most severe form known as lethal hyperphenylalaninemia).

[Phenylketonuria and hyperphenylalaninemia: clinico-genetic classification of 14 forms]. / Fenilketonurii i giperfenilalaninemii: kliniko-geneticheskaia klassifikatsiia 14 form.

The current classifications of the clinical patterns of phenylketonurias and hyperphenylalaninemias are reviewed. On the basis of the literature data, a new classification considering the type of the mutant gene, the severity of the biochemical defect and the disease clinical course is proposed. All 14 hereditary determined patterns are divided into three groups: 1) with complete or partial deficiency of the major enzymes of the reaction of oxidation of phenylalanine into tyrosine; 2) with defects of enzymes in the chain of the cofactor synthesis; 3) with enzymatic defects in conjugated links of phenylalanine and tyrosine metabolism. Some forms are presented in the literature only as rare case reports and the underlying molecular disorders of some of them still remain to be explained. The genetic heterogeneity of phenylketonurias seems to-be much wider than is generally accepted and, therefore, the presented classification can not be considered as final.

[Dihydropteridine reductase activity in leukocytes and cultured fibroblasts of health individuals and of patients with the classical form of phenylketonuria]. / Issledovanie aktivnosti digidropteridinreduktazy v leikotsitakh i kul'tiviruemykh fibroblastakh u zdorovykh liudei i bol'nykh s klassicheskoi formoi fenilketonurii.

Activity of dihydrohypteridine reductase, estimated by means of spectrophotometric method, was found in cell culture of human fibroblasts and in leukocytes of peripheric blood. The enzymatic activity was not altered markedly with time; thus suggesting a possibility to classify dihydropteridine reductase as a constitutive enzyme. In patients with typical form of phenylketonuria the enzymatic activity tended to decrease as compared with healthy persons.

[Genetical heterogeneity of phenylketonuria]. / Geneticheskaia geterogennost' fenilketonurii.

Data on genetic nature of phenylketonuria molecular mechanisms of its pathogenesis and approaches to treatment and prophylaxis of the disease are reviewed. Genetic heterogeneity of phenylketonuria, dependent on polylocus control of phenylalanine hydroxylase complex, is considered in detail. A possibility is discussed of the existence of the genetically different forms of phenylketonuria. Data on the molecular structure of phenylalanine hydroxylase and cooperative nature of its active site are discussed. Variations in pathogenesis of different forms of the phenylketonuria, theoretical and practical significance of these investigations are considered.

[Use of modified Ayling's method for detection of homozygotes and heterozygotes for phenylketonuria gene]. / Primenenie modifitsirovannogo metoda Eiling dlia vyiavleniia gomo- i geterozigot po genu fenilketonurii.

Activity of phenylalanine hydroxylase from human leukocytes was shown to depend on concentration of phenylalanine and pteridine cofactor; optimal concentrations of the substances were estimated. Using this optimized procedure activity of phenylalanine hydroxylase was studied in leukocytes of homo- and heterozygotes by the phenylketonuria gene as well as in the cells of donors. The mean values of the enzymatic activity were distinctly different in the groups of homo-, heterozygote-bearing patients and in healthy persons, although a slight overlapping of the patterns was observed between the groups. Presence of "atypical" forms of phenylketonuria appears to be responsible for this overlapping. The modified procedure for estimation of phenylalanine hydroxylase activity in leukocytes might be used for differential diagnostics of phenylketonuria.

[Presence of active phenylalanine hydroxylase in human leukocytes]. / Dokazatel'stva nalichiia aktivnoi fenilalaningidroksilazy v leikotsitakh cheloveka.

In homogenates of human leukocytes phenylalanine hydroxylase activity was shown to depend on concentration of protein, presence of specific hydroxylase inhibitors as well as on a dose of Ph+ gene; Km value was estimated for phenylalanine and cofactor. The data obtained suggest that human leukocytes contain membrane-bound phenylalanine hydroxylase, which is active in vitro and is similar apparently to the liver enzyme in its properties. Solubilization of the enzyme was achieved without alteration in its activity when 0.25% of Triton X-100 was added by cell homogenisation. Activity of phenylalanine hydroxylase in leukocytes was estimated by a modified procedure of tuling et al. The distinct discrimination between activity of phenylalanine hydroxylase obtained in groups of healthy persons as well as in homo- and heterozygotes by the Ph gene enabled to detect phenylketonuria and to reveal heterozygotes using leukocytes instead of liver tissue biopsy.

[Changes in enzyme activities in early passages of cultured fibroblasts]. / Razlichiia v variatsiiakh aktivnostei fermentov v rannikh passazhakh kul'tiviruemykh fibroblastov.

The specific activities and variability coefficients (Wn) of specific activities of seven enzymes (5 lines) of cultured embryonic human fibroblasts were studied. The experiments were carried out on the 2nd--6th passage (between 12th and 19th passages); in each passage simultaneously up to 3-10 vials collected on the 8th cultivation day were analyzed. According to the Wn value, the enzymes under study can be divided into two groups, i.e. those with a low (less than or equal to 8%) and with a high (25-37%) Wn value. The data obtained are correlated with the division of enzymes and corresponding structural genes into "constituitive" and "inducible" ones. The differences between the enzymes and various lines with respect to the Wn value were revealed. The theoretical and practical significance of the observed changes of the enzyme activities in the cultured cells are discussed.

[Detection of phenylalanine hydroxylase activity in human leukocytes and fibroblasts]. / Obnaruzhenie fenilalaningidroksilaznoi aktivnosti v leikotsitakh i fibroblastakh cheloveka.

The data obtained indicate the presence of phenylalanine hydroxylase activity in human leucocytes and fibroblasts. The following methods were used: estimation of accumulation of the oxidized form of the pteridine cofactor after Ayling and coworkers and radiochemical method. Probably this activity is connected with cell membranes and can be solubilized by treatment of cells by Tritone X-100. The possibility of estimation of phenylalanine hydroxylase activity in leucocytes instead of liver bioptates for discrimination of homo -- and heterozygotes with PKU gene is assumptive.

[Effect of gene dose in human cells]. / Effekt dozy gena v kletkakh cheloveka.

Data from literature are reviewed on the gene dosage effect in human cells. Until recently the dosage effects have been distinctly shown for the genes of superoxide dismutase-1, acid phosphatase of erythrocytes, adenine phosphoribosyl transferase, glutathione reductase and nucleoside phosphorylase, located on chromosomes 21, 2, 16, 8, 14, respectively. The data are discussed in the light of problems arising from the study of the regulation of gene expression in man.

Estimation of correlation of lactate dehydrogenase subunits mole quota based on differences in substrate inhibition.

A kinetic method of estimating the mole quota ratios of the human lactate dehydrogenase (LDH) H and M subunits based on differences in substrate inhibition of LDH isoenzymes by lactate is porposed. Stability of kinetic constants for a prolonged period of time is demonstrated. The dependence of the activity ratios on the contribution of the mole quota of the M-subunit of LDH is studied under conditions of low and high substrate concentrations. The experimental and theoretical values show the following correlation: r = 0.998; p less than 0.001. A comparison of the method proposed with the electrophoretic method of LDH subunit estimation is made, the values obtained being in good agreement. No effect of the components of human diploid cell homogenate and only an insignificant effect of the blood serum components on the kinetic constants of LDH isoenzymes are shown. The applicability of the method to the estimation of the quantitative content of both LDH subunits in natural samples is demonstrated. The informational value of the method is compared to that of other standard methods of LDH isoenzyme estimation.

[Changes in the mole fraction ratio of lactate dehydrogenase subunits in the lymphocytes of Down's syndrome patients]. / Izmenenie sootnosheniia mol'nykh dolei sb''edinits laktatdegidrogenazy v limfotsitakh bol'nykh s sindromom Dauna.

A comparative study of the total activity and mole quota ratio of lactate dehydrogenase subunits in lymphocytes of 14 patients with Down's syndrome (trisomy-21) and in 10 healthy persons is carried out. Differences in the total activity in both groups were insignificant. In patients with Down's syndrome the mole quota ratio of H and M subunits of LDH was found to be significantly altered (p greater than 0.999): H = 33.2%, M - 66,8%, as compared to 51.5% and 48.4% in the control (healthy) group respectively. These differences are evaluated as a result of changed gene expression of both loci controlling H and M polypeptide chains of heteromeric enzyme molecule.

[Comparative study of the changes in lactate dehydrogenase isoenzymatic spectra in the course of organogenesis in mice]. / Sravnitel'noe izuchenie izmenenii izozimnykh spektrov laktatdegidrogenazy v techenie organogeneza u myshei.

The dynamics of changes in the lactate dehydrogenase (LDH) spectra during organogenesis in CBA mice has been studied by means of ultramicroelectrophoresis. The embryonic period of development is characterized by the predominance of cathodic isozymes (LDH-5 and LDH-4) in all the organs under study. The increase of anodic isozymes (LDH-1 and LDH-2) takes place in the heart, kidneys and brain as the development proceeds. The first reliable differences in the LDH spectra of different organs appear on the 11th day of embryogenesis. On the basis of comparison with the help of criterion gamma, the LDH spectra of the organs under study can be divided into two groups: I--heart, lungs, kidneys and brain (tendency towards the increase of H-subunits) and II--intestine, liver and muscles (tendency towards the increase of of M-subunits). The LDH spectra of adult animals are divided into 4 distinct groups: I--heart and kidneys, II--brain, III--lungs and muscles, IV--liver.

[Genetic heterogeneity of G6PD deficiency: mutant alleles of G6PD in the Shekii district of Azerbaijan]. / Geneticheskaia geterogennost' G6FD-nedostatochnosti: issledovanie mutantnykh allelei G6FD v Shekinskom raione Azerbaidzhanskoi SSR.

Examination on G6PD deficiency in 349 patients of Shekii district hospital (Azerbaijan) revealed 16 hemi-, 4 homo- and 9 heterozygotic carriers of the defect. Gd- frequency, calculated from the data obtained (7.7%), may be compared to neighbouring regions' frequencies (6-30%). Carriers of G6PD deficiency are residents of 11 villages located in Alasani-Aphtalan valley, highly endemic with malaria in the past; nearly all marriages are endogamic. Physico-chemical and kinetic study of 10 mutant forms of G6PD, according to WHO program, led to identification of 5 variants of the II class (Shekii, Bideiz, Shirin-Bulakh, Okhut I and Zakataly) and 2 variants of the III class (Okhut II and Martinique-like). Resemblance of the majority of variants in electrophoretic mobility and the level of erythrocyte enzyme activity permit to suggest the existence of a common parental mutant G6PD allele distributed in this area.