A I Glukhov

I.M. Sechenov First Moscow State Medical University, Moskva, Moscow, Russia

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Publications (29)13.76 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Early diagnosis of prostate cancer (CaP) can be addressed by studying prostatic intraepithelial neoplasia (PIN) as precancer (high-grade PIN or HGPIN). This article attempts to analyze the diagnostic role of telomerase as an early marker of carcinogenesis.
    The Prostate 05/2014; · 3.84 Impact Factor
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    ABSTRACT: In order to develop noninvasive diagnostics of bladder cancer (BC), telomerase activity has been examined by means the TRAP method (telomerase repeat amplification protocol) in tumor tissue and urine pellet samples taken from patients with bladder cancer. The levels of relative expression of genes encoding telomerase catalytic subunit (hTERT) and its RNA subunit (hTR) were evaluated by RT-PCR. Telomerase activity and expression of genes encoding its subunits were detected in both tumor tissues and in the urine cell pellet from each BC patient. Results of our study demonstrate possibility of noninvasive BC diagnostics using combination of these methods with sensitivity of 96% and specificity of 100% in the case of telomerase detection and with sensitivity of 80% and specificity of 100% in the case of hTERT detection in urine pellet samples.
    Biochemistry (Moscow) Supplement Series B Biomedical Chemistry 01/2014; 8(1).
  • European Urology Supplements 11/2012; 11(5):193. · 2.16 Impact Factor
  • Zhurnal mikrobiologii, epidemiologii, i immunobiologii 06/2004;
  • Zhurnal mikrobiologii, epidemiologii, i immunobiologii 04/2004;
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    ABSTRACT: The detection of cholera enterotoxin in environmental objects and isolation of patients are considered to be the most reliable indices of that the cholera agent is present. Described in the case study is a method of nested polymerase chain reaction (PCR) based on amplifying the ctxA gene fragment coding subunit A of enterotoxin. A possibility was shown to use the above method to confirm the virulence of strains Vibrio cholerae isolated from different sources. The method was tested with 18 virulent and avirulent strains V. cholerae as well as (for the sake of verifying the analysis specificity) with DNAs of other human-pathogenic microorganisms and with the human genome DNA. The results showed a high efficiency of nested PCR in detecting the pathogenicity of cholera-agent strains.
    Klinicheskaia laboratornaia diagnostika 02/2004;
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    ABSTRACT: To determine the causative agent of hepatitis A in the blood of patients, we developed the method of the nest polymerase chain reaction (PCR) with reverse transcription. For the comparative evaluation of the diagnostic efficacy of the nest PCR and the enzyme immunoassay (EIA) serum samples and mononuclear blood cells obtained from 15 patients with diagnosed hepatitis A and from 33 patients having signs and carrying markers of other form of hepatitis were analyzed. On the whole, the method of PCR confirmed the results of EIA and in some cases exceeded it in sensitivity. In addition, hepatitis A virus RNA was detected in the blood of a clinically healthy person having had a contact with a hepatitis A patient. The PCR data were shown to correlate with the activity of liver amino transferases. The results obtained in this study indicate that the field of the use of PCR in the analysis of hepatitis A virus may include cases of mixed infection and virus carriership.
    Zhurnal mikrobiologii, epidemiologii, i immunobiologii 01/2004;
  • M L Altshuler, S E Severin, A I Glukhov
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    ABSTRACT: Imbalanced activity of the mechanism that controls cell division is a prerequisite for malignant transformation of a normal cell. The present review considers this multi-step mechanism, which is usually called the G1-S checkpoint. Besides, tumor cells are characterized by the presence of telomerase, an enzyme responsible for restoration of chromosome ends after replication and thus providing for unlimited cell division. The main point of the present article is to find out whether the activation of telomerase is controlled by the G1-S checkpoint or does not depend on it. The principal components of the G1-S checkpoint, such as cyclin-dependent kinases, retinoblastoma and E2F proteins, control the activity of telomerase. In their turn, they accumulate and transmit signals from various sources inside and outside the cell. Thus, various changes in tumor cells can activate telomerase through the G1-S checkpoint. Such are the suggested effects on telomerase of Myc, p53, Waf1, protein kinases B and C, Wnt5A, TGFbeta, WT1, and estrogens. However, Myc, p53, WT1, estrogens, protein kinases B and C, and TGFbeta can also directly influence telomerase independently of the G1-S checkpoint mechanism. Moreover, in 30% of human tumors the gene of the key subunit of telomerase (hTERT) is amplified, possibly due to chromosomal rearrangements unassociated with the activity of the G1-S checkpoint. Thus, telomerase seems to be activated not by a single agent but due to combined action of various factors, both with involvement of the G1-S checkpoint mechanism and independently of it.
    Biochemistry (Moscow) 01/2004; 68(12):1275-83. · 1.15 Impact Factor
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    ABSTRACT: The detection of the causative agent of hepatitis A in the patient's body is the necessary element of the diagnostics of this disease. In this work the PCR system for the analysis of the RNA of hepatitis A virus in the patient's blood is presented and characterized. The method of the detection of the RNA of hepatitis A virus is based on the "nest" principle and consists of two consecutive reactions. In the first reaction the reverse transcription and amplification with the external pair of primers are carried out. The product thus obtained is used as material for the second reaction with the internal pair of primers. This method was used for the study of 44 blood samples from hepatitis A patients and 23 blood samples from healthy donors. The detection rate of the RNA of hepatitis A virus in blood samples from the patients was 82%. Viral RNA could be detected in the serum in 72% of cases, both in the serum and in mononuclear blood cells in 20% of cases, in mononuclear blood cells only in 8% of cases.
    Zhurnal mikrobiologii, epidemiologii, i immunobiologii 01/2004;
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    ABSTRACT: Causative agents of plague, i.e. bacterium Yersina pestis (in the subcutaneous tissues of rodents) and their cutaneous parasites need to be isolated to enable plague prevention. A comparatively new method of polymerase chain reaction (PCR) opens up new possibilities of determining Y. pestis just within several hours and without any cultivation. The article contains a description of the PCR-method, which makes it possible to distinguish the culture of Y. pestis from cultures of other microorganism, including speci of Yersina. The method is of the cluster-type, i.e. it is made up of subsequent PC reactions with the substrate for the second reaction being the product of the first one. The cluster nature of the method preconditions a higher sensitivity and specificity versus the ordinary PCR.
    Klinicheskaia laboratornaia diagnostika 08/2003;
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    ABSTRACT: Herpes is one of the most widespread human viral pathologies. The article depicts a special modification of polymerized chain reaction--(PCR)--(referred to as "nested"), which has a higher sensitivity, specificity and reliability as compared to the ordinary PCR, and which is designed to detect the herpes viruses. The method was initially tested at purified preparation of viral DNA, and later--at clinical materials obtained from patients and healthy donors. Secretions from the urogenital tract (smears), scrapes from the urogenital tracts and urinal cellular samples were examined in patients. Herpes simplex was detected in all cases. As for the healthy people, the identical examinations produced in them mainly the negative findings. Therefore, the nested PCR is a simple, sensitive and effective instrument in the diagnostics and prevention of herpes; it can be recommended for a comprehensive usage in medical practice.
    Klinicheskaia laboratornaia diagnostika 03/2003;
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    ABSTRACT: The effect of antisense oligonucleotides complementary to the RNA component of human telomerase on telomerase activity in cell extracts of the melanoma cell line SK-Mel-28 has been studied. It has been shown that the antisense oligonucleotide complementary to the hTR component in the region of the template synthesis of telomeric repeats is the most efficient inhibitor of telomerase activity in comparison with other antisense oligonucleotides. Pronounced inhibition of telomerase activity was observed at the oligonucleotide (Tel P5) concentration in the reaction mixture of about 5 nM. Complete inhibition of the enzyme occurs at the oligonucleotide concentration in the sample of about 20 nM.
    Biochemical and Biophysical Research Communications 08/1998; 248(2):368-71. · 2.41 Impact Factor
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    ABSTRACT: Using thermophilic DNA-polymerase from Thermus thermophilus we have amplified by polymerase chain reaction (PCR) specific DNA sequences of Epstein-Barr virus (EBV) and human immunodeficiency virus (HIV). DNA-polymerase from Thermus thermophilus (molecular mass of 80-86 kDa) differs in its physico-chemical properties from DNA-polymerase from Thermus aquaticus (molecular mass of 62-68 kDa). To amplify the specific EBV DNA sequence, oligonucleotide primers for the virus replicon region (oriP region) were used. As a result of amplification, a specific 405-bp DNA fragment was produced.
    Molecular and Cellular Probes 01/1991; 4(6):435-43. · 1.87 Impact Factor
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    ABSTRACT: The efficiency of phage DNA amplification by the method of polymerase chain reaction (PCR) with Tth DNA-polymerase was studied for optimization of PCR conditions. The effect on amplification efficiency of medium ionic strength and pH, the presence of univalent cations, detergents, gelatin, ATP, pyrophosphate, SH-reagents and ratio of concentrations of Mg and dNTPs, primers and template was studied. It has been found that a pH optimum for PCR with Tth DNA-polymerase varies from 8.5 to 9.0. An ionic strength optimum for PCR is about 0.08. The influence of univalent cations on the activity of Tth DNA-polymerase can be expressed as NH4+ greater than Na+ greater than K+. 0.01% Tween-20 significantly increases the efficiency of PCR and 0.01% gelatin inhibits it. Addition of ATP, pyrophosphate, SH-reagents to the reaction mixture did not increase the yield of PCR product. It has been also shown that for the given PCR-system an optimum Mg/dNTPs molar ratio is within the range of 1.5-2.0. An optimum concentration of each of the pair of primers for this PCR-system is about 0.3 microM. The possibility of PCR-amplification of 500-8500 b.p. DNA fragments has been demonstrated.
    Molekuliarnaia biologiia 01/1991; 25(6):1602-10.
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    ABSTRACT: A thermoresistant htpR mutant having a decreased level of proteolytic activity has been selected in E. coli strain K802 after the directed mutagenesis in vivo. The mutation results in the bacteriophage T7 RNA-polymerase stability, aminoglycosidephosphotransferase stability as well as in the decrease in the rate of proteolytic degradation of cytoplasmic proteins during the heat shock. The obtained mutant strain can, probably be used as a host for alien polypeptides production.
    Molekuliarnaia genetika, mikrobiologiia i virusologiia 10/1988;
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    ABSTRACT: It has been established that the plasmid encoding the N-terminal domain of RecA protein (50 amino acids residues) disturbs SOS functions of htpR- mutants of E. coli. This is expressed in increased UV-sensitivity of strains and in the poor transcription of the aminoglycosidephosphoransferase gene in the hybrid operon recA APT. The discovered effect is due to an increase in the stability of the peptide whose accumulation seems to impede the production of RecA protease. Mutation of gene lon does not lead to the accumulation of N-terminal fragment, therefore an increased stability of the peptide in htpR mutants is probably related to the absence of an unidentified peptidase.
    Molekuliarnaia biologiia 01/1988; 22(5):1198-203.
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    ABSTRACT: The interaction of cAMP-dependent protein kinase from porcine brain with nuclei isolated from the same source was studied. The shape of the curves for the holoenzyme and subunit binding to the nuclei points to the specificity of this interaction and allows for the calculation of the number of protein binding sites. An extremely low degree of binding of the regulatory subunit phosphoform to the nucleus was demonstrated. It was shown that in the nuclei the regulatory subunit binds to proteins whose molecular mass varies from 15,000 and 55,000 Da. Proteolytic fragments of the regulatory subunit do not interact with chromatin proteins; in the nuclei they are functionally inactive and are not involved in protein-protein interactions.
    Biokhimii͡a (Moscow, Russia) 09/1987; 52(8):1300-6.
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    ABSTRACT: The phosphorylation of nuclear proteins of porcine brain cAMP-dependent protein kinase was studied. Some nuclear proteins after extraction from the nuclei served as substrates for protein kinase. Lysine-rich histones H1, H2a and H2b were found to accept phosphate during chromatin phosphorylation by cAMP-dependent protein kinase. Phosphorylation of intact nuclei revealed that in such a system only histone H1 is a substrate for cAMP-dependent protein kinase. In the presence of DNA the histones are phosphorylated by cAMP-dependent protein kinase in a different manner. It was concluded that DNA can determine the accessibility of protein substrates for the catalytic subunit of cAMP-dependent protein kinase.
    Biokhimii͡a (Moscow, Russia) 08/1987; 52(7):1150-3.
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    ABSTRACT: The effect of the catalytic and regulatory subunits of cAMP-dependent protein kinase type II on development, proliferation, and RNA synthesis was studied in loach embryos. It was found that injection of the catalytic subunit in a physiological concentration leads to a disturbance in the course of development and inhibits proliferation and RNA synthesis in the embryos. An increase in the concentration of this protein above the physiological level leads to death of the embryos in the first hours of development. Injection of the regulatory subunit stimulated the incorporation of labeled uridine into the acid-insoluble fraction of the embryos, beginning with the gastrula stage. The cell nuclei of loach embryos injected with subunits of protein kinase type II were transplanted into activated loach egg cells: subunits of protein kinase type I had no effect on the ability of nuclei of undetermined loach embryo cells to provide de novo development and their effect was reversible.
    06/1986; 50:12.
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    ABSTRACT: Using the method of protein transfer from polyacrylamide gel to nitrocellulose filters with subsequent incubation of filter-adsorbed protein with [32P]DNA, it was found that the catalytic subunit of cAMP-dependent protein kinase from porcine brain is capable of interacting with DNA to form a stable complex. This complex is resistant even to 2 M NaCl. The ability of the catalytic subunit to interact with DNA depends on the degree of enzyme nativity. The regulatory subunit of cAMP-dependent protein kinase does not bind to DNA both in the presence and absence of cAMP. The 125I-labeled regulatory subunit can interact with some chromatin proteins, in particular, with histone H1 and core histones. An essential role in this binding belongs to electrostatic and hydrophobic interactions.
    Biokhimii͡a (Moscow, Russia) 02/1986; 51(1):103-11.

Publication Stats

61 Citations
13.76 Total Impact Points

Institutions

  • 2014
    • I.M. Sechenov First Moscow State Medical University
      Moskva, Moscow, Russia
  • 1991
    • Russian Research Center for Molecular Diagnostics and Therapy
      Московский, Moskovskaya, Russia
  • 1982
    • Институт молекулярной биологии им. В.А. Энгельгардта Российской академии наук
      Moskva, Moscow, Russia