Enzymatic synthesis of structure-free DNA with pseudo-complementary properties

Long single-stranded DNAs and RNAs possess considerable secondary structure under conditions that support stable hybrid formation with oligonucleotides. Consequently, different oligomeric probes can hybridize to the same target with efficiencies that vary by several orders of magnitude. The ability to enzymatically generate structure-free singlestranded copies of any nucleic acid without impairing Watson–Crick base pairing to short probes would eliminate this problem and significantly improve the performance of many oligonucleotide-based applications. Synthetic nucleic acids that exhibit these properties are defined as pseudo-complementary. Previously, we described a pseudo-complementary A-T couple consisting of 2-aminoadenine (nA) and 2-thiothymine (sT) bases. The nA-sT couple is a mismatch even though nA-T and A-sT are stable base pairs. Here we show that 7-alkyl-7-deazaguanine and N 4 -alkylcytosine (where alkyl = methyl or ethyl) can be used in conjunction with nA and sT to render DNA largely structure-free and pseudo-complementary. The deoxynucleoside triphosphates (dNTPs) of these bases are incorporated into DNA by selected mesophilic and thermophilic DNA polymerases and the resulting primer extension products hybridize with good specificity and stability to oligonucleotide probes composed of the standard bases. Further optimization and characterization of the synthesis and properties of pseudo-complementary DNA should lead to an ideal target for use with oligonucleotide probes that are <25 nt in lengthThis work was supported by grants from the National Institutes of Health (NIGMS Grant 74564 to H.G.) and Agilent Technologies. Dr Carl Fuller (GE Healthcare) kindly provided us with dNTPs of 7-alkyl-7-deazaguanines and 7-iodo-7-deazaguanine. We thank Dr Jeffrey Sampson (Agilent Technologies) for his constant encouragement and Caryn Evila (Idaho State University) for her early participation in the project. At TriLink Biotechnologies we are grateful to Dr Richard Hogrefe, Dr Gerald Zon, Dr Natasha Paul and Dr David Combs for support and helpful discussions and Dr Inna Koukhareva and Stephanie Perry for synthesis of dNTPs. Funding to pay the Open Access publication charges for this article was provided by NIH GM74564.Peer reviewe

On the ability of indentation to capture the anisotropic elastic properties of the pyrolytic carbon

We used micro- and nanoindentation, macroscopic strain gage methods and FE studies to correlate the indentation modulus and the macroscopic in- and out-of-plane Young\u27s moduli of transversely isotropic pyrolytic carbon. The out-of-plane indentation modulus of bulk PyroC was comparable with predicted values and more than a factor of two higher than the out-of-plane Young\u27s modulus. The in-plane indentation modulus was significantly lower than predicted values which we attribute to nanobuckling of graphite-like planes. Polarized light microscopy studies on carbon-carbon composite samples infiltrated at 10, 20, 30 kPa demonstrated that the increase in pressure leads to lower texture of the PyroC and higher in-plane indentation modulus due to a decreased tendency toward nanobuckling for more isotropic structures. We observed damage of PyroC layers at critical loads when indenting normal to the plane of isotropy and attribute the damage to a combination of crack propagation and delamination types of deformation mechanisms

Enzymatic synthesis of structure-free DNA with pseudo-complementary properties.

Long single-stranded DNAs and RNAs possess considerable secondary structure under conditions that support stable hybrid formation with oligonucleotides. Consequently, different oligomeric probes can hybridize to the same target with efficiencies that vary by several orders of magnitude. The ability to enzymatically generate structure-free single-stranded copies of any nucleic acid without impairing Watson-Crick base pairing to short probes would eliminate this problem and significantly improve the performance of many oligonucleotide-based applications. Synthetic nucleic acids that exhibit these properties are defined as pseudo-complementary. Previously, we described a pseudo-complementary A-T couple consisting of 2-aminoadenine (nA) and 2-thiothymine (sT) bases. The nA-sT couple is a mismatch even though nA-T and A-sT are stable base pairs. Here we show that 7-alkyl-7-deazaguanine and N(4)-alkylcytosine (where alkyl = methyl or ethyl) can be used in conjunction with nA and sT to render DNA largely structure-free and pseudo-complementary. The deoxynucleoside triphosphates (dNTPs) of these bases are incorporated into DNA by selected mesophilic and thermophilic DNA polymerases and the resulting primer extension products hybridize with good specificity and stability to oligonucleotide probes composed of the standard bases. Further optimization and characterization of the synthesis and properties of pseudo-complementary DNA should lead to an ideal target for use with oligonucleotide probes that arelength

THE MODERN VIEWS TO DIAGNOSIS OF KIDNEY DAMAGE IN PATIENTS WITH ARTERIAL HYPERTENSION FOR EARLY PREVENTION

The article discusses in detail the problem of the medico-social significance of early kidney damage in essential hypertension (EH) (according to the literature review and own research). Aim. To determine the prevalence of subclinical renal damage in the EAG of the first stage and to assess the diagnostic significance of the functional renal reserve (FRR). Materials and methods. A total of 70 patients were examined. The control group consisted of 50 people with established diagnosis of EH I stage, women - 20 (40%), mean age – 44,6±1,78 years, men - 30 (60%), average age – 46,57±2,23 years. The average duration of the EH is 3,5 ± 0,5 years. The control group included 20 practically healthy persons (11 women (55%), men - 9 (45%), average age 41,1 ± 1,96 years). All the patients underwent the following examinations: clinical examination, general clinical blood and urine tests, biochemical blood test (total protein, urea, creatinine, glucose, lipidogram) 24-hour BP monitoring, ECG, ECHO-CG, carotid ultrasound, definition FRR. The body mass index (BMI), the glomeru-laorpfhiltthratlimonosrcaotpey(aGnFdR) was calculated. Results. According to the FRR, the functional state of the kidneys significantly differs between subgroups of patients with EH stage I (p <0.05). In patients with the control group FRR - 62.14±10.60%, compared with patients with EH of the 1st subgroup – 23,31±4,9% and the 2nd subgroup - 10±3,02%. In patients of the 2nd subgroup of FRR, although within normal limits, it is noteworthy that it is located on the lower border. When performing the correlation analysis of the data of FRR and BP, the dependence was inversely proportional (r = -0.7, p <0.05). Conclusion. The eGFR was determined by the CKD-EPI formula in the examined patients with EH and control groups, which does not differ significantly. FRR in patients with EH I-II grade with the duration of the disease up to 5 years is significantly lower than in practically healthy peers; p <0.05. In patients with EH II grade (2nd subgroup) FNR more than twice as low as patients with EAG I degree; and is 10±3,2% and 23,3±4,9% respectively; p <0.05. Thus, according to the results of the study, it was found that FRR is a more sensitive indicator, which can help in the early diagnosis of kidney damage with EH

PHENOMENOLOGY OF VALUES OF THE NOVEL OF INGARDEN

Статья рассматривает инструментарий феноменологии как ценностно- смысловой модели описания социокультурной динамики. Предмет рассматривается в связи с юбилеем кончины Романа Ингардена, ученика Гуссерля. Особый интерес представляют модификации смысловой деятельности, спайка интерсубъективности, рождение коллективных жизненных миров. Социокультурная феноменология исходит из того, что множество трансцендентальных субъектов конституируют мир культуры.The paper considers the tools of phenomenology as a value-semantic model for describing sociocultural dynamics. The subject is considered in connection with the anniversary of the death of Roman Ingarden, a student of Husserl. Of particular interest are modifications of semantic activity, a spike of intersubjectivity, the birth of collective life worlds. Sociocultural phenomenology proceeds from the fact that many transcendental subjects constitute the world of culture

POST-COVID-19 SUSTAINABILITY ISSUES

The paper is devoted to the social aspects of counteracting the spread of the disease, which challenged global economic relations, brought down global markets and undermined national economies. The purpose of the article is to consider a set of measures to preserve social sustainability. Research methods: comparative, genetic and factors types of analyzes. Results: the topic is presented in the context of the concept of a post-modern state, where sovereignty depends on multiple international and domestic factors, including oligarchical elites. Research novelty: global pandemic situation has forced to rethink the issues of economic security, optimized medicine, global factors of regional sustainability. Viruses as a nano factor on the scale of our macro universe have actualized the synergetic teachings on the antinomic connection between the living and nonliving, global and regional, scientific and mythological. When at school, we were taught about four traditional kingdoms of lifе, i.e. animals, plants, fungi and bacteria. The recent advent of the coronavirus pandemic made humanity face the fifth kingdom, the viruses. This paper is devoted to research of legal instruments containing both administrative and criminal law measures intent to organize the society in times of the pandemic. Within a few weeks, the novelty virus COVID-19 has spread worldwide by crossing national borders and sparking hot debates about national and global biosafety hazards. Conclusion: the research maintains that liberal models of public management can work effectively only where normal life conditions and law-abiding population can accept regular methods of governance. The world of work is different to many of us in COVID times and this change is global. For engineering professionals, working in VUCA world remote working i.e. work from home is today’s reality. Remote working, remote meeting, remote decision-making, remote control, and at the same time ensuring productivity is the scenario most of the professionals are currently passing through. Too much virtual interaction with no fixed working hours is impacting mental health. There is a need for a work life balance too. Social life will be most affected while working from home permanently. Once the ongoing pandemic situation gets into control, there may be an option of allowing employee to work from home if need be. Also, everybody cannot work from home. Too much of seclusion and no face to face interaction with peers and colleagues can have a negative impact on individuals particularly mental stress would increase. The HR model would now need to focus on Well-being as DNA of the company. The remote working should also have defined start and stop time. Healthy mind overall sustains the healthy organization. Austerity and Pragmatism is a way to go ahead. It is a total IT spin therefore HR's role is more on trust development.The paper is devoted to the social aspects of counteracting the spread of the disease, which challenged global economic relations, brought down global markets and undermined national economies. The purpose of the article is to consider a set of measures to preserve social sustainability. Research methods: comparative, genetic and factors types of analyzes. Results: the topic is presented in the context of the concept of a post-modern state, where sovereignty depends on multiple international and domestic factors, including oligarchical elites. Research novelty: global pandemic situation has forced to rethink the issues of economic security, optimized medicine, global factors of regional sustainability. Viruses as a nano factor on the scale of our macro universe have actualized the synergetic teachings on the antinomic connection between the living and nonliving, global and regional, scientific and mythological. When at school, we were taught about four traditional kingdoms of lifе, i.e. animals, plants, fungi and bacteria. The recent advent of the coronavirus pandemic made humanity face the fifth kingdom, the viruses. This paper is devoted to research of legal instruments containing both administrative and criminal law measures intent to organize the society in times of the pandemic. Within a few weeks, the novelty virus COVID-19 has spread worldwide by crossing national borders and sparking hot debates about national and global biosafety hazards. Conclusion: the research maintains that liberal models of public management can work effectively only where normal life conditions and law-abiding population can accept regular methods of governance. The world of work is different to many of us in COVID times and this change is global. For engineering professionals, working in VUCA world remote working i.e. work from home is today’s reality. Remote working, remote meeting, remote decision-making, remote control, and at the same time ensuring productivity is the scenario most of the professionals are currently passing through. Too much virtual interaction with no fixed working hours is impacting mental health. There is a need for a work life balance too. Social life will be most affected while working from home permanently. Once the ongoing pandemic situation gets into control, there may be an option of allowing employee to work from home if need be. Also, everybody cannot work from home. Too much of seclusion and no face to face interaction with peers and colleagues can have a negative impact on individuals particularly mental stress would increase. The HR model would now need to focus on Well-being as DNA of the company. The remote working should also have defined start and stop time. Healthy mind overall sustains the healthy organization. Austerity and Pragmatism is a way to go ahead. It is a total IT spin therefore HR's role is more on trust development

Social Policy in the Changing Architecture of the Welfare State: What Will Remain of Costly Soviet Social Rights?

Soviet society contributed to the growth of workers’ social rights. The right to housing, the right to free education and free medical care, the right to rest and leisure — all these are expensive social rights that were established as a social standard in the USSR and contributed to the improvement of the position of the working man around the world. Oligarchic state capitalism means replacing expensive social rights with “cheap” political rights.Советское общество способствовало росту социальных прав трудящихся. Право на жилье, право на бесплатное образование и бесплатное медицинское обслуживание, право на отдых — все это дорогие социальные права, которые были установлены как социальный стандарт в СССР и содействовали улучшению положения человека труда во всем мире. Олигархический государственный капитализм означает замену дорогих социальных прав на «дешевые» политические права

EFFECT OF PRIORITY DRINKING WATER CONTAMINANTS ON THE QUALITY INDICATORS OF BEVERAGES DURING THEIR PRODUCTION AND STORAGE

Currently, water from the centralized domestic drinking water supply system is mainly used to make non- alcoholic carbonated beverages and nectars. The classical technology does not always provide the purification of water from organic compounds. In addition, during water preparation, at the primary chlorination stage, chlorine-containing organic compounds (chloroform, dichloroethane, trichlorethylene, etc.) are formed due to the interaction of chlorine with natural organic substances. The by-products of natural water treatment by chloragents, in addition to the toxic and carcinogenic effects, can interact with the main components of products reducing their quality. Such water cannot be used for drinking purposes and in food production without the additional post-treatment. The results of the study of the effect of organic impurities present in water (chloroform, trichlorethylene and dichloroethane) on the stability of the components of non-alcoholic carbonated beverages (sodium benzoate, sucrose, citric acid, natural and synthetic dyes and vanillin) and nectars (color stability, vitamins A, C, group B) have been provided. The studies were carried out in the Kemerovo region using gas-liquid chromatography, molecular absorption spectroscopy, refractometry and capillary electrophoresis. The concentration of the main components of non-alcoholic carbonated beverages, nectars and priority contaminants (trichlorethylene and dichloroethane) has been reduced. It has been shown that chloroform does not come into chemical interaction with the components of beverages. The mechanism of interaction of sucrose, citric acid, sodium benzoate, vanillin, vitamins in non-alcoholic carbonated beverages and nectars with trichlorethylene and dichloroethane has been theoretically justified. It has been established that dichloroethane and trichlorethylene have a significant effect on the resistance of the main components of non-alcoholic carbonated beverages, with the exception of dyes, and also on the intensity of color and the preservation of vitamins of nectars, reducing the quality characteristics of beverages during production and storage

Genetic control of N-glycosylation of human blood plasma proteins

Glycosylation is an important protein modification, which influences the physical and chemical properties as well as biological function of these proteins. Large-scale population studies have shown that the levels of various plasma protein N-glycans are associated with many multifactorial human diseases. Observed associations between protein glycosylation levels and human diseases have led to the conclusion that N-glycans can be considered a potential source of biomarkers and therapeutic targets. Although biochemical pathways of glycosylation are well studied, the understanding of the mechanisms underlying general and tissue-specific regulation of these biochemical reactions in vivo is limited. This complicates both the interpretation of the observed associations between protein glycosylation levels and human diseases, and the development of glycan-based biomarkers and therapeutics. By the beginning of the 2010s, high-throughput methods of N-glycome profiling had become available, allowing research into the genetic control of N-glycosylation using quantitative genetics methods, including genome-wide association studies (GWAS). Application of these methods has made it possible to find previously unknown regulators of N-glycosylation and expanded the understanding of the role of N-glycans in the control of multifactorial diseases and human complex traits. The present review considers the current knowledge of the genetic control of variability in the levels of N-glycosylation of plasma proteins in human populations. It briefly describes the most popular physical-chemical methods of N-glycome profiling and the databases that contain genes involved in the biosynthesis of N-glycans. It also reviews the results of studies of environmental and genetic factors contributing to the variability of N-glycans as well as the mapping results of the genomic loci of N-glycans by GWAS. The results of functional in vitro and in silico studies are described. The review summarizes the current progress in human glycogenomics and suggests possible directions for further research