Proton Distribution Radii of

The root mean square radii of the proton density distribution in ^{16-24}O derived from measurements of charge changing cross sections with a carbon target at ∼900A MeV together with the matter radii portray thick neutron skin for ^{22-24}O despite ^{22,24}O being doubly magic. Imprints of the shell closures at N=14 and 16 are reflected in local minima of their proton radii that provide evidence for the tensor interaction causing them. The radii agree with ab initio calculations employing the chiral NNLO_{sat} interaction, though skin thickness predictions are challenged. Shell model predictions agree well with the data.

A Method for Assessing the Efficiency of the Nucleotide Excision Repair System Ex Vivo.

The nucleotide excision repair (NER) is one of the main repair systems present in the cells of living organisms. It is responsible for the removal of a wide range of bulky DNA lesions. We succeeded in developing a method for assessing the efficiency of NER in the cell (ex vivo), which is a method based on the recovery of TagRFP fluorescent protein production through repair of the damage that blocks the expression of the appropriate gene. Our constructed plasmids containing bulky nFlu or nAnt lesions near the tagrfp gene promoter were shown to undergo repair in eukaryotic cells (HEK 293T) and that they can be used to analyze the efficiency of NER ex vivo. A comparative analysis of the time dependence of fluorescent cells accumulation after transfection with nFlu- and nAnt-DNA revealed that there are differences in how efficient their repair by the NER system of HEK 293T cells can be. The method can be used to assess the cell repair status and the repair efficiency of different structural damages.

[Application of GLAD-PCR Assay for Study on DNA Methylation in Regulatory Regions of Some Tumor-Suppressor Genes in Lung Cancer].

Hypermethylation of the gene regulatory regions are common for many cancer diseases. In this work we applied GLAD-PCR assay for identificating of the aberrantly methylated RCGY sites in the regulatory regions of some downregulated genes in tissue samples of lung cancer (LC). This list includes EFEMP1, EPHA5, HOXA5, HOXA9, LHX1, MYF6, NID2, OTX1, PAX9, RARB, RASSF1A, RXRG, SIX6, SKOR1 and TERT genes. The results of DNA samples from 40 cancer and 25 normal lung tissues showed a good diagnostic potential of selected RCGY sites in regulatory regions of MYF6, SIX6, RXRG, LHX1, RASSF1A and TERT genes with relatively high sensitivity (80.0 %) and specificity (88.0 %) of LC detection in tumor DNA.

New Experimental Models of Retinal Degeneration for Screening Molecular Photochromic Ion Channel Blockers.

Application of molecular photochromic ion channel blockers to recover the visual function of a degenerated retina is one of the promising trends in photopharmacology. To this day, several photochromic azobenzene-based compounds have been proposed and their functionality has been demonstrated on cell lines and knockout mouse models. Further advance necessitates testing of the physiological activity of a great number of new compounds. The goal of this study is to propose animal models of photoreceptor degeneration that are easier to obtain than knockout mouse models but include the main features required for testing the physiological activity of molecular photoswitches. Two amphibian-based models were proposed. The first model was obtained by mechanical deletion of the photoreceptor outer segments. The second model was obtained by intraocular injection of tunicamycin to induce the degeneration of rods and cones. To test our models, we used 2-[(4-{(E)-[4-(acryloylaminophenyl]diazenyl}phenyl)amino]-N,N,N-triethyl-2-oxoethanammonium chloride (AAQ), one of the compounds that have been studied in other physiological models. The electroretinograms recorded from our models before and after AAQ treatment are in agreement with the results obtained on knockout mouse models and reported in other studies. Hence, the proposed models can be used for primary screening of molecular photochromic ion channel blockers.

[DNA Bearing Bulky Fluorescent and Photoreactive Damage in Both Strands as Substrates of the Nucleotide Excision Repair System].

Model DNA molecules that contain bulky lesions in both strands have been created, and their properties as substrates of the nucleotide excision repair (NER) system have been analyzed. The modified nucleoside, 5-[3-(4-azido-2,3,5,6-tetrafluorobenzamido)-1-propoxypropyl]-2'-deoxycytidine (dC^(FAB)), or the nonnucleoside fragment, N-[6-(9-anthracenylcarbamoyl)hexanoyl]-3-amino-1,2-propanediol (nAnt), have been inserted as damage in certain positions of the first DNA strand ("0"). The position of N-[6-5(6)-fluoresceinylcarbamoyl]hexanoyl] -3-amino-1,2-propanediol (nFlu) has been varied within the second DNA strand. This residue has been located opposite the removable damaging fragment of the first strand at positions -20, -10, -4, 0, +3, and +8 relative to the first lesion. It has been demonstrated that the presence of nFlu at the -4, 0, or +3 position of the second strand significantly reduces the thermostability of DNA duplexes, especially in the case of nAnt-DNA and completely excludes the possibility of NER-catalyzed excision from dC^(FAB)- and nAnt-containing 137-meric DNA with the second lesion at these positions. The introduction of nFlu at positions -20, -10, or +8 differently affects the excision efficiency of dC^(FAB)- and nAnt-containing fragments from the first strand. The excision efficiency of dC^(FAB)-containing fragments from extended double-damaged DNA is as high as from DNA that contains a single dC^(FAB) damage, while the excision of nAnt-containing fragments occurs with 80-90% lower efficiency from double-damaged DNA occurs from DNA that contains the single nAnt insert. The nFlu insert differently affects the interaction of the sensory XPC-HR23B dimer with dC^(FAB)- and nAnt-containing DNAs, although in all cases, this interaction occurs with increased efficiency compared to that with single-damaged DNAs. No direct correlation between the thermostability of the DNA duplex and XPC-DNA affinity on the one hand, and the excision efficiency of lesions on the other hand has been shown. The absence of the correlation may be caused by both functional features of variable multiprotein complexes involved in the recognition and verification of damage during NER and the sensitivity of the complexes to the structure of the damage and damage-surrounding DNA. The results are important for understanding the NER mechanism of elimination of bulky damage located in both DNA strands.

Genome Stability Maintenance in Naked Mole-Rat.

The naked mole-rat (Heterocephalus glaber) is one of the most promising models used to study genome maintenance systems, including the effective repair of damage to DNA. The naked mole-rat is the longest lived rodent species, which is extraordinarily resistant to cancer and has a number of other unique phenotypic traits. For at least 80% of its lifespan, this animal shows no signs of aging or any increased likelihood of death and retains the ability to reproduce. The naked mole-rat draws the heightened attention of researchers who study the molecular basis of lengthy lifespan and cancer resistance. Despite the fact that the naked mole-rat lives under genotoxic stress conditions (oxidative, etc.), the main characteristics of its genome and proteome are a high stability and effective functioning. Replicative senescence in the somatic cells of naked mole-rats is missing, while an additional p53/pRb-dependent mechanism of early contact inhibition has been revealed in its fibroblasts, which controls cell proliferation and its mechanism of arf-dependent aging. The unique traits of phenotypic and molecular adaptations found in the naked mole-rat speak to a high stability and effective functioning of the molecular machinery that counteract damage accumulation in its genome. This review analyzes existing results in the study of the molecular basis of longevity and high cancer resistance in naked mole-rats.

Proton Distribution Radii of

Proton radii of ^{12-19}C densities derived from first accurate charge changing cross section measurements at 900A MeV with a carbon target are reported. A thick neutron surface evolves from ∼0.5 fm in ^{15}C to ∼1 fm in ^{19}C. The halo radius in ^{19}C is found to be 6.4±0.7 fm as large as ^{11}Li. Ab initio calculations based on chiral nucleon-nucleon and three-nucleon forces reproduce the radii well.

First Measurement of Several ß-Delayed Neutron Emitting Isotopes Beyond N=126.

The ß-delayed neutron emission probabilities of neutron rich Hg and Tl nuclei have been measured together with ß-decay half-lives for 20 isotopes of Au, Hg, Tl, Pb, and Bi in the mass region N≳126. These are the heaviest species where neutron emission has been observed so far. These measurements provide key information to evaluate the performance of nuclear microscopic and phenomenological models in reproducing the high-energy part of the ß-decay strength distribution. This provides important constraints on global theoretical models currently used in r-process nucleosynthesis.

DNA with Damage in Both Strands as Affinity Probes and Nucleotide Excision Repair Substrates.

Nucleotide excision repair (NER) is a multistep process of recognition and elimination of a wide spectrum of damages that cause significant distortions in DNA structure, such as UV-induced damage and bulky chemical adducts. A series of model DNAs containing new bulky fluoro-azidobenzoyl photoactive lesion dC(FAB) and well-recognized nonnucleoside lesions nFlu and nAnt have been designed and their interaction with repair proteins investigated. We demonstrate that modified DNA duplexes dC(FAB)/dG (probe I), dC(FAB)/nFlu+4 (probe II), and dC(FAB)/nFlu-3 (probe III) have increased (as compared to unmodified DNA, umDNA) structure-dependent affinity for XPC-HR23B (Kdum > KdI > KdII ≈ KdIII) and differentially crosslink to XPC and proteins of NER-competent extracts. The presence of dC(FAB) results in (i) decreased melting temperature (ΔTm = -3°C) and (ii) 12° DNA bending. The extended dC(FAB)/dG-DNA (137 bp) was demonstrated to be an effective NER substrate. Lack of correlation between the affinity to XPC-HR23B and substrate properties of the model DNA suggests a high impact of the verification stage on the overall NER process. In addition, DNAs containing closely positioned, well-recognized lesions in the complementary strands represent hardly repairable (dC(FAB)/nFlu+4, dC(FAB)/nFlu-3) or irreparable (nFlu/nFlu+4, nFlu/nFlu-3, nAnt/nFlu+4, nAnt/nFlu-3) structures. Our data provide evidence that the NER system of higher eukaryotes recognizes and eliminates damaged DNA fragments on a multi-criterion basis.

[Application of GLAD-PCR analysis for the methylation sites detection in the regulatory areas of tumor-suppressor genes ELMo1 and EsR1 in colorectal cancer].

Aberrant methylation of regulation regions of tumorsuppressor genes is showed for many cancer diseases. In course of this modification an enzyme DNMT3 methylates RCGY sites in CpG-islands of regulation regions producing R(5mC)GY sites. Earlier we developed GLAD-PCR assay to determine R(5mC)GY site in a definite position of human genome. In this work we have applied GLAD-PCR assay to determine R(5mC)GY sites in regulation regions of ESR1 and ELMO1 tumor-suppressor genes. We have studied a fragment of first exon of ELMO1 gene and a part of ESR1 promoter region in DNA preparations from malignant cell line SW837 and colorectal tumor samples. We have checked four sites in each region and found two highly methylated sites: GCGC in first exon of ELMO1 gene and GCGT in promoter region of ESR1 gene. Site GCGT is weakly methylated in healthy tissues and more methylated in the most of colorectal samples. Site GCGC is not methylated in healthy tissues and significantly methylated in 60% of colorectal samples. A possibility to use GLAD-PCR assay for cancer diagnostics is discussed.

[DISTRIBUTION OF TWO-LOCUS HAPLOTYPES OF MICROBIAL COMPONENT SENSOR GENES TLR1 AND TLR6 IN MAJOR POPULATIONS OF SOUTH URALS].

AIM: Evaluate the fraction of various TLR1-TLR6 haplotypes in populations of Russians Bashkir and Nagaybak of Chelyabinsk Region. MATERIALS AND METHODS: Potential donors of stem cells from Chelyabinsk Region Station of Blood Transfusion registry were included into the study and split into 3 populations: Russians (81), Bashkir (78) and Nagaybak (84). Genotyping by 2 polymorphisms of TLR1 and TLR6 genes was carried out in all the 3 groups. Point polymorphism of TLR1 gene 1805T>G was determined by polymorphism analysis of length of restriction fragments, and polymorphism of TLR6 gene 745C>T by PCR using sequence-specific primers. RESULTS: TLR1 1805*G-TLR6 745*T haplotype occurs in population of Russians (42%) and relatively rare--among Bashkir (17%). An inverse picture is observed for TLR1 1805*T-TLR6 745*C haplotype: a more frequent spread among Bashkir (65%) and relatively rare occurrence in Russians (23%). Frequencies of the mentioned haplotypes, that occupy intermediate position compared with corresponding parameters for populations of Russians and Bashkir, were detected for Nagaybak, that, probably, reflects complex pathways of settling of their ancestors and effects of other non-adaptation factors. CONCLUSION: Frequencies of TLR1-TLR6 two-locus haplotypes in major populations of South Urals were determined for the first time. Further studies in this field will allow better understanding of features of immune response and sensitivity to infections in various populations.

[Clinical and ultrasonic predictors of postoperative complications after carotid endarterectomy].

For the period from 2007 to 2012 carotid endarterectomy was performed in 150 patients with cerebrovascular insufficiency I-IV degrees and atherosclerotic lesion of carotid arteries. Dynamic observation was performed by using of duplex scanning of brachiocephalic arteries, transcranial duplex scanning, multislice CT with contrast study of extracranial and intracranial arteries. Different degrees of vascular wall thickening of operated internal carotid artery including neo- and myointimal hyperplasia, restenosis and other complications were observed in 19 (12.6%) patients after carotid endarterectomy on background of cerebrovascular insufficiency progressing. It was revealed that transient ischemic attack or stroke, acute heart failure in early postoperative period, arterial hypertension with crisis course predominantly, diabetes mellitus 2 type, obesity, male sex, elderly age and smoking were clinical markers for complications after carotid endarterectomy. Ultrasonic markers of complications after carotid endarterectomy included terms of development and degree of vascular wall thickening in case of neointimal hyperplasia and restenosis, hyperperfusion syndrome and stroke, significant changes of blood flow velocity and indexes of peripheral vascular resistance.

[Inflammatory diseases of scrotal organs in patients with brucellosis: Improvement of therapy].

AIM: To evaluate the efficacy of cycloferon used in the combination treatment of scrotal inflammatory diseases (SID) in patients with brucellosis. SUBJECTS AND METHODS: A total of 150 male patients with chronic brucellosis (CB) were examined. Inquiry, questioning, physical and ultrasound examinations, and spermiogram analysis were used to detect of diseases of the reproductive system. Twenty-two CB patients with SID were examined over time (before and after cycle therapy). In Group 1, combination therapy included 2 cycles of five intramuscular injections of cycloferon 0.25 g in each at a 10-day interval. In Group 2, a package of therapeutic measures meets the generally accepted standards. and Incorporation of cycloferon into the combination therapy of SID patients with CB positively affected the time course of clinical changes and spermatogenesis, declines the number of SID exacerbations, improved quality of life, and failed to cause side effects. CONCLUSION: The findings allow us to recommend cycloferon as the drug of choice in treating CB patients with SID.

Proton radii of (12-17)B define a thick neutron surface in ¹7B.

The first determination of radii of point proton distribution (proton radii) of (12-17)B from charge-changing cross sections (σ(CC)) measurements at the FRS, GSI, Darmstadt is reported. The proton radii are deduced from a finite-range Glauber model analysis of the σ(CC). The radii show an increase from ¹³B to ¹7B and are consistent with predictions from the antisymmetrized molecular dynamics model for the neutron-rich nuclei. The measurements show the existence of a thick neutron surface with neutron-proton radius difference of 0.51(0.11) fm in ¹7B.

Molecular mechanism of global genome nucleotide excision repair.

Nucleotide excision repair (NER) is a multistep process that recognizes and eliminates a wide spectrum of damage causing significant distortions in the DNA structure, such as UV-induced damage and bulky chemical adducts. The consequences of defective NER are apparent in the clinical symptoms of individuals affected by three disorders associated with reduced NER capacities: xeroderma pigmentosum (XP), Cockayne syndrome (CS), and trichothiodystrophy (TTD). These disorders have in common increased sensitivity to UV irradiation, greatly elevated cancer incidence (XP), and multi-system immunological and neurological disorders. The eucaryotic NER system eliminates DNA damage by the excision of 24-32 nt single-strand oligonucleotides from a damaged strand, followed by restoration of an intact double helix by DNA repair synthesis and DNA ligation. About 30 core polypeptides are involved in the entire repair process. NER consists of two pathways distinct in initial damage sensor proteins: transcription-coupled repair (TC-NER) and global genome repair (GG-NER). The article reviews current knowledge on the molecular mechanisms underlying damage recognition and its elimination from mammalian DNA.

[Design of deoxyribozymes for inhibition of influenza A virus].

Influenza A viruses take a significant place in human and animal pathology causing epidemics and epizootics. Therefore, the development of new antiflu drugs has become more and more urgent. Deoxyribozymes can be considered as promising antiviral agents due to their ability to efficiently and highly specifically cleave RNA molecules. In this study, a number ofgenomic sequences of the most relevant influenza A virus subtypes, H5N1, H3N2, and H1N1, were analyzed. Conservative regions were revealed in five the least variable segments of the fragmented viral RNA genome, and potential sites of their cleavage with "10-23" deoxyribozymes were determined. 46 virus-specific 33-mer deoxyribozymes with the general structure of 5'N8AGGCTAGCTACAACGAN9 were designed and synthesized. Screening of the antiviral activity of these agents in conjugation with lipofectin on the Madin-Darby Canine Kidney cells infected with highly pathogenic avian influenza virus A/chicken/Kurgan/05/2005 (H5N1) revealed 17 deoxyribozymes, which suppressed the titer of virus cytopathicity by more than 2.5 IgTCID50/mL (i.e. the virus neutralization index was more than 300), with five of them suppressing the virus titer by a factor of 1000 and more. The most active deoxyribozymes appeared to be specific to segment 5 of the influenza A virus genome, which encoded nucleoprotein (NP).

[Surgical treatment of hemodynamically significant kinking of internal carotid arteries].

38 patients with different forms of vascular cerebral insufficiency, caused by kinking and atherosclerotic changes of internal carotid arteries were operated on. Various types of reconstructive operation on extracranial carotid arteries were performed. The color duplex ultrasound scanning and computed tomography proved to be highly informative noninvasive means for detecting carotid pathology in patients with vascular cerebral insufficiency. Reconstructive operations on internal carotid arteries can serve as prophylactic and treatment measure of chronic cerebral insufficiency. Authors propose the principle of "six types" of reconstructive operations which individualizes the surgical approach. Carotid surgery for asympomatic patients should be performed on strict indications.

[Effectiveness of cycloferon in complex treatment of brucellosis patients with lesions of the scrotum].

The article presents the results of urological examination (spermograms and data of ultrasound examination) of 22 patients with chronic brucellosis and diseases of the scrotum (6 patients with orchitis, 16 with orchiepididymitis) before and after conventional therapy (10 patients) and combined treatment with the inclusion of cycloferon (2 courses of 5 intramuscular injection [0.25 g] with an interval of 10 days)--12 patients. It is shown that the administration of cycloferon leads to more effective relief of intoxication symptoms and inflammation in the testes and appendages (reduction of scrotal wall thickness, size of testes and/or adjuncts, and the incidence and severity of hydrocele), and has a positive effect on spermatogenesis (reduction of semen viscosity, the number of white blood cells in semen, sperm agglutination associated with the formation of sperm antibodies in most patients after treatment), as well as reduces the number of exacerbations of chronic orchitis/orchiepididymitis by 2.4 times.