End invasion of peptide nucleic acids (PNAs) with mixed-base composition into linear DNA duplexes

Peptide nucleic acid (PNA) is a synthetic DNA mimic with valuable properties and a rapidly growing scope of applications. With the exception of recently introduced pseudocomplementary PNAs, binding of common PNA oligomers to target sites located inside linear double-stranded DNAs (dsDNAs) is essentially restricted to homopurine–homopyrimidine sequence motifs, which significantly hampers some of the PNA applications. Here, we suggest an approach to bypass this limitation of common PNAs. We demonstrate that PNA with mixed composition of ordinary nucleobases is capable of sequence-specific targeting of complementary dsDNA sites if they are located at the very termini of DNA duplex. We then show that such targeting makes it possible to perform capturing of designated dsDNA fragments via the DNA-bound biotinylated PNA as well as to signal the presence of a specific dsDNA sequence, in the case a PNA beacon is employed. We also examine the PNA–DNA conjugate and prove that it can initiate the primer-extension reaction starting from the duplex DNA termini when a DNA polymerase with the strand-displacement ability is used. We thus conclude that recognition of duplex DNA by mixed-base PNAs via the end invasion has a promising potential for site-specific and sequence-unrestricted DNA manipulation and detection.National Institutes of Health (CA74175, GM059173); Boston University (PIF and SPRING awards

End invasion of peptide nucleic acids (PNAs) with mixed-base composition into linear DNA duplexes

Peptide nucleic acid (PNA) is a synthetic DNA mimic with valuable properties and a rapidly growing scope of applications. With the exception of recently introduced pseudocomplementary PNAs, binding of common PNA oligomers to target sites located inside linear double-stranded DNAs (dsDNAs) is essentially restricted to homopurine–homopyrimidine sequence motifs, which significantly hampers some of the PNA applications. Here, we suggest an approach to bypass this limitation of common PNAs. We demonstrate that PNA with mixed composition of ordinary nucleobases is capable of sequence-specific targeting of complementary dsDNA sites if they are located at the very termini of DNA duplex. We then show that such targeting makes it possible to perform capturing of designated dsDNA fragments via the DNA-bound biotinylated PNA as well as to signal the presence of a specific dsDNA sequence, in the case a PNA beacon is employed. We also examine the PNA–DNA conjugate and prove that it can initiate the primer-extension reaction starting from the duplex DNA termini when a DNA polymerase with the strand-displacement ability is used. We thus conclude that recognition of duplex DNA by mixed-base PNAs via the end invasion has a promising potential for site-specific and sequence-unrestricted DNA manipulation and detection.National Institutes of Health (CA74175, GM059173); Boston University (PIF and SPRING awards

Soft X-Ray Sources at the Centers of the Elliptical Galaxies NGC 4472 and NGC 4649

Analysis of recent Chandra observations of the elliptical galaxies NGC 4472 and NGC 4649 has revealed faint soft X-ray sources at their centers. The sources are located to within 1'' of the optical centers of the galaxies. They are most likely associated with the central supermassive black holes. Interest in these and several other similar objects stems from the unusually low luminosity of the supermassive black holes embedded in dense interstellar medium. Our Chandra sources have very soft spectra. They are detectable only below ~0.6 keV and have luminosities in the 0.2-2.5 keV energy band of ~ 6 * 10^{37} erg/s and ~1.7 * 10^{38} erg/s in NGC 4649 and NGC 4472, respectively.Comment: Shortened version of the paper published in Astronomy Letter

ПРОФИЛАКТИКА ПОСТТРАНСФУЗИОННОГО ВИРУСНОГО ГЕПАТИТА С ПРИ ПЕРЕЛИВАНИИ ДОНОРСКОЙ КРОВИ И ЕЕ КОМПОНЕНТОВ

The aim of organizational aspects of preventing the transmission of hepatitis C virus with donor blood and its components.Materials and methods. An activity of the blood service establishments in Russia for the prevention of HCV infection through transfusion of blood and its components on the basis of the analysis of sectoral statistical surveys was studied.Results. The frequency of detection of antibodies to hepatitis C virus in blood donors and its components during 2009–2013 decreased by more than 1,5 times. The percentage of donors who have identified markers of hepatitis C virus was significantly different in different regions: from 0,51% to 1,36%. The activity of the blood service implemented method of plasma quarantine resulting annually rejected from 0,32% to 0,23% as a result of the identified markers of HCV. Pathogen inactivated plasma volume increased in 3 times, the platelet concentrate in 3,2 times.Conclusion. To ensure the safety of donated blood and its components in the blood service effectively the modern technology use for to prevention transmission of the HCV: quarantine of plasma, donor selection and development, inactivation of pathogens. The degree of implementation in practice of nonpaid voluntary blood transfusions significantly increased and is characterized by regional features in recent years .Цель. Исследование организационных аспектов профилактики передачи вирусного гепатита С с донорской кровью и ее компонентами.Материалы и методы. Проведено изучение деятельности учреждений службы крови России по предупреждению инфицирования ВГС при переливании донорской крови и ее компонентов на основе анализа отраслевых статистических наблюдений.Результаты. Частота выявления антител к вирусному гепатиту С у доноров крови и ее компонентов в течение 2009–2013 гг. снизилась более чем в 1,5 раз. Процентное число доноров, у которых выявлены маркеры вируса гепатита С, существенно отличалось в различных регионах: от 0,51% до 1,36%. В деятельность службы крови внедрен метод карантинизации плазмы, в результате чего ежегодно бракуется от 0,32% до 0,23% вследствие выявленных маркеров ВГС. Объем патогенинактивированной плазмы увеличился в 3 раза, тромбоцитного концентрата – в 3,2 раза.Заключение. Для обеспечения безопасности донорской крови и ее компонентов в службе крови эффективно применяются современные технологии профилактики передачи ВГС: карантинизация плазмы, отбор доноров и развитие безвозмездного добровольного донорства, инактивация патогенов. Cтепень их внедрения в трансфузиологическую практику в последние годы существенно повышается и характеризуется региональными особенностями

MYB suppresses differentiation and apoptosis of human breast cancer cells

Introduction: MYB is highly expressed in estrogen receptor positive (ER + ve) breast tumours and tumour cell lines. We recently demonstrated that MYB is essential for the proliferation of ER + ve breast cancer cells, and have now investigated its role in mammary epithelial differentiation.Methods: MCF-7 breast cancer cells were treated with sodium butyrate, vitamin E succinate or 12-O-tetradecanoylphorbol-13-acetate to induce differentiation as measured by Nile Red staining of lipid droplets and β-casein expression. The non-tumorigenic murine mammary epithelial cell (MEC) line, HC11, was induced to differentiate with lactogenic hormones. MYB levels were manipulated by inducible lentiviral shRNA-mediated knockdown and retroviral overexpression.Results: We found that MYB expression decreases following chemically-induced differentiation of the human breast cancer cell line MCF-7, and hormonally-induced differentiation of a non-tumorigenic murine mammary epithelial cell (MEC) line, HC11. We also found that shRNA-mediated MYB knockdown initiated differentiation of breast cancer cells, and greatly sensitised them to the differentiative and pro-apoptotic effects of differentiation-inducing agents (DIAs). Sensitisation to the pro-apoptotic effects DIAs is mediated by decreased expression of BCL2, which we show here is a direct MYB target in breast cancer cells. Conversely, enforced expression of MYB resulted in the cells remaining in an undifferentiated state, with concomitant suppression of apoptosis, in the presence of DIAs.Conclusions: Taken together, these data imply that MYB function is critical in regulating the balance between proliferation, differentiation, and apoptosis in MECs. Moreover, our findings suggest MYB may be a viable therapeutic target in breast cancer and suggest specific approaches for exploiting this possibility

The possible functions of duplicated ets (GGAA) motifs located near transcription start sites of various human genes

Transcription is one of the most fundamental nuclear functions and is an enzyme complex-mediated reaction that converts DNA sequences into mRNA. Analyzing DNA sequences of 5′-flanking regions of several human genes that respond to 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in HL-60 cells, we have identified that the ets (GGAA) motifs are duplicated, overlapped, or clustered within a 500-bp distance from the most 5′-upstream region of the cDNA. Multiple protein factors including Ets family proteins are known to recognize and bind to the GGAA containing sequences. In addition, it has been reported that the ets motifs play important roles in regulation of various promoters. Here, we propose a molecular mechanism, defined by the presence of duplication and multiplication of the GGAA motifs, that is responsible for the initiation of transcription of several genes and for the recruitment of binding proteins to the transcription start site (TSS) of TATA-less promoters

PARP-1 and YY1 Are Important Novel Regulators of CXCL12 Gene Transcription in Rat Pancreatic Beta Cells

Despite significant progress, the molecular mechanisms responsible for pancreatic beta cell depletion and development of diabetes remain poorly defined. At present, there is no preventive measure against diabetes. The positive impact of CXCL12 expression on the pancreatic beta cell prosurvival phenotype initiated this study. Our aim was to provide novel insight into the regulation of rat CXCL12 gene (Cxcl12) transcription. The roles of poly(ADP-ribose) polymerase-1 (PARP-1) and transcription factor Yin Yang 1 (YY1) in Cxcl12 transcription were studied by examining their in vitro and in vivo binding affinities for the Cxcl12 promoter in a pancreatic beta cell line by the electrophoretic mobility shift assay and chromatin immunoprecipitation. The regulatory activities of PARP-1 and YY1 were assessed in transfection experiments using a reporter vector with a Cxcl12 promoter sequence driving luciferase gene expression. Experimental evidence for PARP-1 and YY1 revealed their trans-acting potential, wherein PARP-1 displayed an inhibitory, and YY1 a strong activating effect on Cxcl12 transcription. Streptozotocin (STZ)-induced general toxicity in pancreatic beta cells was followed by changes in Cxcl12 promoter regulation. PARP-1 binding to the Cxcl12 promoter during basal and in STZ-compromised conditions led us to conclude that PARP-1 regulates constitutive Cxcl12 expression. During the early stage of oxidative stress, YY1 exhibited less affinity toward the Cxcl12 promoter while PARP-1 displayed strong binding. These interactions were accompanied by Cxcl12 downregulation. In the later stages of oxidative stress and intensive pancreatic beta cell injury, YY1 was highly expressed and firmly bound to Cxcl12 promoter in contrast to PARP-1. These interactions resulted in higher Cxcl12 expression. The observed ability of PARP-1 to downregulate, and of YY1 to upregulate Cxcl12 promoter activity anticipates corresponding effects in the natural context where the functional interplay of these proteins could finely balance Cxcl12 transcription