Evaluation of antiviral activity of terpenophenols and some of their N- and O-derivatives.

A comparative evaluation of the antiviral activity of a number of new and previously synthesized terpenophenols and their N- or O-containing derivatives against the A/Puerto Rico/8/34 (H1N1) virus strain was carried out. 2-Isobornylphenol, 1,2-dihydroxy-6-isobornyl-4-methylbenzene, 2-isobornyl-1,4-benzoquinone, and N-butyl-4-hydroxy-3,5-diisobornylbenzamide showed the highest activity.

Triethylamine-Based Salts: Protic Ionic Liquids or Molecular Complexes?

Amines can interact with protic acids with different degrees of proton transfer, which can lead to the formation of both hydrogen-bonded complexes and protic ionic liquids (PILs) in which the hydrogen bond between the cation and anion contributes to the formation of ion pairs. This work is devoted to studying the degree of proton transfer from different acids (hydrochloric, nitric, phosphoric, acetic, propionic, benzoic, and salicylic) to triethylamine (TEA). The results of quantum-chemical calculations based on the density functional theory (DFT) and thermal (phase transition and destruction temperatures) and physicochemical (conductivity, viscosity) characteristics of the compounds show that TEA interaction with acetic and propionic acids leads to the formation of hydrogen-bonded complexes. The B3LYP-GD3 method also shows that the interaction between TEA and benzoic acid is more energetically favorable for the formation of a molecular complex, whereas the obtained experimental data are more characteristic of a protic ionic liquid. For the other acids studied, the calculation and experimental data confirm salt formation. The geometric and energy parameters of the H-bond have been calculated both in the molecular complexes and in the ion pairs. The QTAIM theory was used to localize critical points of the hydrogen bonds and to calculate their properties.

[Control levels of Sin3 histone deacetylase for spontaneous and UV-induced mutagenesis in yeasts Saccharomyces cerevisiae].

SIN3 gene product operates as a repressor for a huge amount of genes in Saccharomyces cerevisiae. Sin3 protein with a mass of about 175 kDa is a member of the RPD3 protein complex with an assessed mass of greater than 2 million Da. It was previously shownthat RPD3 gene mutations influence recombination and repair processes in S. cerevisiae yeasts. We studied the impacts of the sin3 mutation on UV-light sensitivity and UV-induced mutagenesis in budding yeast cells. The deletion ofthe SIN3 gene causes weak UV-sensitivity of mutant budding cells as compared to the wild-type strain. These results show that the sin3 mutation decreases both spontaneous and UV-induced levels of levels. This fact is hypothetically related to themalfunction of ribonucleotide reductase activity regulation, which leads to a decrease in the dNTP pool and the inaccurate error-prone damage bypass postreplication repair pathway, which in turn provokes a reduction in the incidence of mutations.

[Interaction of the HSM3 gene with genes initiating homologous recombination repair in yeast Saccharomyces cerevisiae].

It was assumed previously that the mutator phenotype of the hms3 mutant was determined by processes taking place in the D-loop. As a next step, genetic analysis was performed to study the interactions between the hsm3 mutation and mutations of the genes that control the initial steps of the D-loop formation. The mutations of the MMS4 and XRS2 genes, which initiate the double-strand break formation and subsequent repair, were shown to completely block HSM3-dependent UV-induced mutagenesis. Mutations of the RAD51, RAD52, and RAD54 genes, which are also involved in the D-loop formation, only slightly decreased the level of UV-induced mutagenesis in the hsm3 mutant. Similar results were observed for the interaction of hsm3 with the mph1 mutation, which stabilizes the D-loop. In contrast, the shu1 mutation, which destabilizes the D-loop structure, led to an extremely high level of UV-induced mutagenesis and displayed epistatic interactions with the hsm3 mutation. The results made it possible to assume that the hsm3 mutation destabilizes the D-loop, which is a key substrate of both Rad5- and Rad52-dependent postreplicative repair pathways.

[Interaction of gene HSM3 with genes of the epistatic RAD6 group in yeast Saccharomyces cerevisiae].

In eukaryotes, damage tolerance of matrix DNA is mainly determined by the repair pathway under the control of the RAD6 epistatic group of genes. T this pathway is also a main source of mutations generated by mutagenic factors. The results of our recent studies show that gene HSM3 participating in the control of adaptive mutagenesis increases the frequency of mutations induced by different mutagens. Mutations rad18, rev3, and mms2 controlling various stages of the RAD6 pathway are epistatic with mutation hsm3 that decreases UV-induced mutagenesis to the level typical for single radiation-sensitive mutants. The level of mutagenesis in the double mutant srs2 hsm3 was lower than in both single mutants. Note that a decrease in the level of mutagenesis relative to the single mutant srs2 depends on the mismatch repair, since this level in the triple mutant srs2 hsm3 pms 1 corresponds to that in the single mutant srs2. These data show that the mutator phenotype hsm3 is probably determined by processes occurring in a D loop. In a number of current works, the protein Hsm3 was shown to participate in the assembly of the proteasome complex S26. The assembly of proteasomes is governed by the N-terminal domain. Our results demonstrated that the Hsm3 protein contains at least two domains; the N-terminal part of the domain is responsible for the proteasome assembly, whereas the C-terminal portion of the protein is responsible for mutagenesis.

[Surface active properties of isobornylphenols in systems with different degrees of complexity].

Interrelations between the structure of the semi-synthetic phenolic antioxidants -- isobornylphenols and their surface active properties were studied in the chemical (the lecithin aggregation in hexane) and biological (the incubation with the blood erythrocytes) model systems. It has been shown that all studied compounds are able to affect the lecithin aggregation in hexane: the share of the main fraction of the L micelles decreases with increasing the share of particles of greater size. The effect substantially depends on hindered OH group and the presence of the intramolecular hydrogen bond in molecule. The cytotoxic properties of isobornylphenols (the concentration is 100 M) are predominantly due to the molecule structure. The interrelation between the aggregate size of the main fraction of L in the presence of the studied compounds and the discocyte share during mice blood erythrocyte incubation in their presence for 4 h is revealed. Thus, this provides the possibility to assume that the ability of the different biological active substances to affect the lecithin aggregation in non-polar solvent could be used as a model system for the initial assessment of their surface active properties.

[Genetic analysis of the Hsm3 protein domain structure in yeast Saccharomyces cerevisiae].

Gene HSM3 encodes the Hsm3 protein involved in the minor branch in the system responsible for the correction of mismatched bases in DNA structure and controls replicative and reparative spontaneous mutagenesis in yeast Saccharomyces cerevisiae. Spontaneous and UV-induced mutagenesis was studied in three mutant alleles of gene HSM3, and repair effectivity of artificial heteroduplexes was assessed in DNA molecule. The resuts of these studies allowed establishment of the protein domain structure of protein Hsm3 and functions of each domain: the N-terminal domain is responsible for binding to mispaired bases, and the C-terminal domain ensures the interaction with other proteins involved in the system of mismatched base correction.

[The geptrong pharmaceutical product increases efficiency of postreplication repair of permutation intermediates in yeast Saccharomyces cerevisiae].

Geptrong is a medication from pure defermentated honey. In medical practice, it is used as hepatoprotector. Genotoxicity analysis revealed antimutagenic activity of the preparation. The spontaneous mutation rate at the ADE4-ADE8 and CAN1 loci was several times lower in case that the yeast cells were plated on the geptrong-containing medium, and the mutation rate was scored using the method of ordered plating. If spontaneous mutation rate was measured by means of the fluctuation method of median, no antimutagenic activity was detected. Geptrong had no effect on the yeast cell survival. At the same time, it substantially decreased the frequency of direct mutations at the ADE4-ADE8 locus, induced by UV- and gamma-radiation, and ethylmetansulfonate. The effect of the geptrong antimutagenic activity on the level of UV-induced mutagenesis in the yeast strains defective for the repair systems rad2, rad51, rad54, rad59, msh2, and hsm3 was examined. Antimutagenic activity was detected in the wild type, as well as in the rad2, rad54, rad59, and hsm3 strains, while rad51, pmsl, and msh2 mutants lacked this activity. Based on these data, it is suggested that antimutagenic effect of geptrong is associated with activated repair of mismatches, appeared during the postreplicative recombination repair.

Requirement of HSM3 gene for spontaneous mutagenesis in Saccharomyces cerevisiae.

In this work, we studied the influence of hsm3 mutation on spontaneous mutagenesis in actively and slowly dividing cells. We demonstrated that the spontaneous mutation rates in the hsm3 mutant and the wild type strain were similar in actively dividing cells. However, during 15-day cultivation of both strains we observed higher accumulation of mutants in the hsm3 strain compared with those in the wild type cells. Effect of accumulation of spontaneous mutants was observed in slowly dividing cells in the rad1, rad2, rad14, rad54, and pms1, but it was absent in the rev3, pol2 and pol3 mutants. Combinations of the hsm3 mutation with the pol3-01, pol2-04 and pms1Delta mutations decreased significantly the level of spontaneous mutagenesis in rapidly growing cells. The hsm3 mutation suppressed synthetic lethality in the hsm3 pol3-01 pms1 triple mutant and dramatically increased the spontaneous mutation rate in comparison with double mutant. The introduction of the hsm3 mutation in NER-mutants led to considerably increasing of the spontaneous mutation level. The double hsm3 rev3, hsm3 rad54 and hsm3 pms1Delta mutants showed lower spontaneous mutation rate compared with the single mutants in rapidly dividing cells. The combination the hsm3 mutation with all studied mutations characterized by different degree of increase of spontaneous mutagenesis in slowly dividing cells. The participation of the Hsm3p in spontaneous mutagenesis in slowly and activity dividing yeast cells is discussed.

HSM2 (HMO1) gene participates in mutagenesis control in yeast Saccharomyces cerevisiae.

We have previously reported about a new Saccharomyces cerevisiae mutation, hsm2-1, that results in increase of both spontaneous and UV-induced mutation frequencies but does not alter UV-sensitivity. Now HSM2 gene has been genetically and physically mapped and identified as a gene previously characterized as HMO1, a yeast homologue of human high mobility group genes HMG1/2. We found that hsm2 mutant is slightly deficient in plasmid-borne mismatch repair. We tested UV-induced mutagenesis in double mutants carrying hsm2-1 mutation and a mutation in a gene of principal damaged DNA repair pathways (rad2 and rev3) or in a mismatch repair gene (pms1 and recently characterized in our laboratory hsm3). The frequency of UV-induced mutations in hsm2 rev3 was not altered in comparison with single rev3 mutant. In contrast, the interaction of hsm2-1 with rad2 and pms1 was characterized by an increased frequency of UV-induced mutations in comparison with single rad2 and pms1 mutants. The UV-induced mutation frequency in double hsm2 hsm3 mutant was lower than in the single hsm2 and hsm3 mutants. The role of the HSM2 gene product in control of mutagenesis is discussed.

The yeast HSM3 gene acts in one of the mismatch repair pathways.

Mutants with enhanced spontaneous mutability (hsm) to canavanine resistance were induced by N-methyl-N-nitrosourea in Saccharomyces cerevisiae. One bearing the hsm3-1 mutation was used for this study. This mutation does not increase sensitivity to the lethal action of different mutagens. The hsm3-1 mutation produces a mutator phenotype, enhancing the rates of spontaneous mutation to canavanine resistance and reversions of lys1-1 and his1-7. This mutation increases the rate of intragenic mitotic recombination at the ADE2 gene. The ability of the hsm3 mutant to correct DNA heteroduplex is reduced in comparison with the wild-type strain. All these phenotypes are similar to ones caused by pms1, mlhl and msh2 mutations. In contrast to these mutations, hsm3-1 increases the frequency of ade mutations induced by 6-HAP and UV light. Epistasis analysis of double mutants shows that the PMS1 and HSM3 genes control different mismatch repair systems. The HSM3 gene maps to the right arm of chromosome II, 25 cM distal to the HIS7 gene. Strains that bear a deleted open reading frame YBR272c have the genetic properties of the hsm3 mutant. The HSM3 product shows weak similarity to predicted products of the yeast MSH genes (homologs of the Escherichia coli mutS gene). The HSM3 gene may be a member of the yeast MutS homolog family, but its function in DNA metabolism differs from the functions of other yeast MutS homologs.

[Mutator genes of the yeast Saccharomyces cerevisiae. Interaction of mutations him and his with mutations blocking three principal pathways of repair of induced DNA damage]. / Geny-mutatory drozhzhei Saccharomyces cerevisiae. Vzaimodeistvie mutatsii him i hsm s mutatsiiami, blokiruiushchimi tri osnovnykh puti reparatsii indutsirovannykh povrezhdenii DNK.

During recent years, genes controlling mutation in higher eukaryotes have been found to be involved actively in carcinoma regeneration in cells. In this respect, studying the genetic control of mutagenesis becomes a key direction of research into mechanisms responsible for cancer generation. The results of studying interaction of mutations in the HIM and HSM genes, controlling spontaneous and induced mutagenesis in yeasts, and mutations impairing three known pathways of DNA damage repair in this microorganism, are described in this work. It was shown that mutation rev3 completely blocks UV-induced mutagenesis in all mutants studied. On the other hand, mutation rad2 synergistically interacts with mutations him1, hsm1, hsm3, hsm6, and hsm2, thus enhancing the frequency of UV-induced mutagenesis in double mutants multiple times. Mutations him2 and him3 manifested epistatic interaction with mutation rad2. With mutation rad54, the interaction was epistatic for mutations him1 and hsm2 and was additive for mutations hsm1, him2, and him3. On the basis of the data obtained, we developed a scheme for the appearance of mismatch bases in the process of repair of UV-induced DNA damage.

[Mutator genes from Saccharomyces cerevisiae. Repair of artificial heteroduplexes in him and hsm mutants]. / Geny-mutatory drozhzhei Saccharomyces cerevisiae. Reparatsiia iskusstvennykh geterodupleksov u mutantov him i hsm.

During recent years, genes controlling mutation in higher eukaryotes have been found to be involved actively in carcinoma regeneration in cells. In this respect, studying the genetic control of mutagenesis becomes a key direction of research into mechanisms responsible for cancer generation. The results of studying interaction of mutations in the HSM3 and HSM6 genes, controlling spontaneous and induced mutagenesis in yeasts, and mutations impairing three known pathways of DNA damage repair in this microorganism, are described in this work. It was shown that mutation rev3 completely blocks UV-induced mutagenesis in all mutants studied. On the other hand, mutation rad2 synergistically interacts with mutations him1, hsm1, hsm3, hsm6, and hsm2, thus enhancing the frequency of UV-induced mutagenesis in double mutants multiple times. Mutations him2 and him3 manifested epistatic interaction with mutation rad2. With mutation rad54, the interaction was epistatic for mutations him1 and hsm2 and was additive for mutations hsm1, him2, and him3. On the basis of the data obtained, we developed a scheme for the appearance of mismatch bases in the process of repair of UV-induced DNA damage.

[Mutator genes from Saccharomyces cerevisiae. Interaction between HIM- and HSM-genes]. / Geny-mutatory drozhzhei Saccharomyces cerevisiae. Vzaimodeistvie mezhdu HIM- i HSM-genami.

The interaction of six mutator genes of the yeast Saccharomyces cerevisiae with respect to UV-induced mutagenesis was studied. To this effect, double mutants with a genotype containing pairs of mutations at genes analyzed were synthesized. Analysis of the type of interaction of these mutations revealed four epistatic gene groups: (1) HIM1, HSM3, and HSM6; (2) HSM1; (3) HSM2; and (4) HIM2 and HIM3. The possible role of genes studied in different repair pathways of mispaired bases was discussed.

[Experience in using serological tests in detecting tuberculosis in patients with severe mental pathology]. / Opyt ispol'zovaniia serologicheskikh testov v vyiavlenii tuberkuleza u bol'nykh s tiazheloi psikhicheskoi patologiei.

113 patients with severe mental diseases were examined for tuberculosis using enzyme immunoassay which provided positive or doubtful results in 15% of the examinees. At clinicoroentgenological examination the patients were found to have active respiratory tuberculosis, foci of indefinite activity and posttuberculous changes in the lungs and pleura in 4.4, 3.5 and 7.1%, respectively.

[Effect of hms mutations increasing spontaneous mutability on induced mutagenesis and mitotic recombination in the yeast Saccharomyces cerevisiae]. / Vliianie mutatsii hms, povyshaiushchikh spontannuiu mutabil'nost', na industsirovannyi mutagenez i mitotcheskuiu rekombinatsiiu u drozhzhei Saccharomyces cerevisiae.

The influence of five nonallelic mutations hsm-1-hsm-5 on the frequency of mutations induced by UV-light, 6-hydroxyl-aminopurine (GAP) and nitrosomethylurea (NMM) at the ADE1 and ADE2 loci was studied. All hsm mutants were resistant to the lethal effect of these mutagens. The frequency of mutations induced by UV-light was increased in hsm2-1, hsm3-1, hsm5-1 and especially in hsm1-1 mutants, the hsm4-1 mutant not differing from the HSM strain. GAP-induced mutagenesis was elevated in all hsm mutants and, particularly, in hsm3-1. No influence of hsm mutations on the frequency of NMM-induced mutations was observed. The frequency of spontaneous mitotic gene conversion was studied in the diploids heteroallelic for mutations in the gene ADE2 (ade2-58 ade2-i) and homo- and heterozygous for the hsm mutations (HSMHSM and HSMhsm). The mutations hsm2-1, hsm3-1 and especially hsm5-1 strongly increased the conversion frequency for all heteroallelic combinations studied. The mutations hsm1-1, hsm4-1 affected this process weakly. The properties of the hsm mutations under study demonstrated common genetic control of spontaneous and induced mutagenesis and recombination in the yeast. Possible belonging of hsm mutations to the class of mutations destroying the repair pathway for mismatch correction is under discussion.

[Isolation and characteristics of new mutants of Saccharomyces cerevisiae with increased spontaneous mutability]. / Vydelenie i kharakteristika novykh mutantov drozhzhei Saccharomyces cerevisiae s povyshennoi spontannoi mutabil'nost'iu.

To isolate some new genes controlling the process of spontaneous mutagenesis, a collection of 16 yeast strains with enhanced rate of spontaneous canavanine resistant mutations was obtained. Genetical analysis allowed to define that the mutator phenotype of these strains is due to a single nuclear mutation. Such mutations were called hsm (high spontaneous mutagenesis). Recombinational test showed that 5 mutants under study carried 5 nonallelic mutations. It was revealed that the mutation hsm3-1 is a nonspecific mutator elevating the rate of both spontaneous canavanine resistant mutations and the frequency of reversions in mutations lys1-1 and his1-7. Genetical analysis revealed that mutation hsm3-1 is recessive. The study of cross sensitivity of mutator strains to physical and chemical mutagens demonstrated that 12 of 16 hsm mutants were resistant to the lethal action of UV, gamma rays and methylmethanesulfonate, and 4 mutants were only sensitive to these factors. Possible nature of hsm mutations is discussed.

[Changes in the lactate dehydrogenase isoenzyme activity in parturients with chronic fetal hypoxia during different types of anesthesia in surgical delivery]. / Izmeneniia aktivnosti izofermentov laktatdegidrogenazy u rozhenits s khronicheskoi gipoksiei ploda pri razlichnykh vidakh anestezii v usloviiakh operativnogo rodorazresheniia.

During cesarean delivery, the isoenzymic spectrum of lactate dehydrogenase and lactic acid levels were examined in 109 parturients with chronic fetal hypoxia. For initial anesthesia, sombrevin, kalipsol, hexenal were applied, basal anesthesia was performed with neuroleptic analgetics and combined electric anesthesia. The findings make it possible to choose the optimal anesthetic agents in this group of parturients in order to make the status of a fetus and a neonate stable and better.

[Mutagenesis in cloned yeast genes. The effect of mutation rad2 on the frequency of gene mutation in plasmid and chromosome]. / Mutagenez na klonirovannykh drozhzhevykh genakh. Vliianie mutatsii rad2 na chastotu mutirovaniia gena v sostave plazmidy i khromosomy.

The influence of rad2 mutation blocking incision of pyrimidine dimers on frequency of UV-light and 6-hydroxylaminopurine (6-GAP)-induced adenine-independent revertants was studied in the strains of Saccharomyces cerevisiae containing the same mutant allele of gene ADE2 in episomic plasmid and in chromosome. It was shown that the strains carrying the ade2 mutation in chromosome and in plasmid did not differ in sensitivity to lethal action of UV-light and 6-GAP. However, in the plasmid rad2 strain reversions were induced by UV-light more frequently (approximately 100 times), as compared to the chromosome strain. We observed no significant differences between reversion frequencies in plasmid and chromosome RAD strains. The tendency to enhanced 6-GAP-induced mutagenesis, less sharply expressed, was observed in the chromosome rad2 strain, as compared to the plasmid one. However, the plasmid RAD strain was characteristic of higher reversion frequency induced by 6-GAP, as compared to the chromosome strain. The possible mechanisms of these phenomena are discussed.