A DNA assembly toolkit to unlock the CRISPR/Cas9 potential for metabolic engineering

CRISPR/Cas9-based technologies are revolutionising the way we engineer microbial cells. One of the key advantages of CRISPR in strain design is that it enables chromosomal integration of marker-free DNA, eliminating laborious and often inefficient marker recovery procedures. Despite the benefits, assembling CRISPR/Cas9 editing systems is still not a straightforward process, which may prevent its use and applications. In this work, we have identified some of the main limitations of current Cas9 toolkits and designed improvements with the goal of making CRISPR technologies easier to access and implement. These include 1) A system to quickly switch between marker-free and marker-based integration constructs using both a Cre-expressing and standard Escherichia coli strains, 2) the ability to redirect multigene integration cassettes into alternative genomic loci via Golden Gate-based exchange of homology arms, 3) a rapid, simple in-vivo method to assembly guide RNA sequences via recombineering between Cas9-helper plasmids and single oligonucleotides. We combine these methodologies with well-established technologies into a comprehensive toolkit for efficient metabolic engineering using CRISPR/Cas9. As a proof of concept, we developed the YaliCraft toolkit for Yarrowia lipolytica, which is composed of a basic set of 147 plasmids and 7 modules with different purposes. We used the toolkit to generate and characterize a library of 137 promoters and to build a de novo strain synthetizing 373.8 mg/L homogentisic acid

Gold nanoparticles - the theranostic challenge for PPPM: nanocardiology application

The article overviews the potential biomedical applications of nanoscale gold particles for predictive, preventive and personalised nanomedicine in cardiology. The review demonstrates the wide opportunities for gold nanoparticles due to their unique biological properties. The use of gold nanoparticles in cardiology is promising to develop fundamentally new methods of diagnosis and treatment. The nanotheranostics in cardiovascular diseases allows the non-invasive imaging associated with simultaneous therapeutic intervention and predicting treatment outcomes. Imaging may reflect the effectiveness of treatment and has become a fundamental optimisation setting for therapeutic protocol. Combining the application of biomolecular and cellular therapies with nanotechnologies foresees the development of complex integrated nanodevices. Nanocardiology may challenge existing healthcare system and economic benefits as cardiovascular diseases are the leading cause of morbidity and mortality at present

МОДЕЛЮВАННЯ МЕТОДИК РЕГІОНАРНОЇ АНЕСТЕЗІЇ ПІД УЛЬТРАЗВУКОВИМ КОНТРОЛЕМ НА ФАНТОМАХ З ЗАСТОСУВАННЯМ МЕТОДІВ СТАТИСТИЧНОГО АНАЛІЗУ

The paper compares different techniques of needle insertion during phantom modeling of regional anesthesiaaffecting ultrasound visualization. A statistical analysis of experimental data for methodics optimization was conducted.В статье сравниваются разные способы введения иглы при региональной анестезии, влияющие на ультразвуковую визуализацию. Проведен статистико-математический анализ полученных в результате эксперимента данных.В статті порівнюються різні способи введення голки при регіонарній анестезії, що впливають на ультразвукову візуалізацію. Проведено статистико-математичний аналіз отриманих у результаті експерименту даних

Prophylactic effect of lactobacilli and bifidobacteria probiotic strains on experimental bacterial vaginitis

The objective of the study was determining the prophylactic effect of Lactobacillus casei IMV B-7280, L. acidophilus IMV B-7279, L. delbrueckii subsp. bulgaricus IMV B-7281, Bifidobacterium animalis VKL and B. animalis VKB probiotic strains on experimental vaginitis in BALB/c mice induced by Staphylococcus aureus 8325-4. The infection with S. aureus 8325-4 caused an imbalance of microbiota in the vagina and intestine, as evidenced by an increase in the number of opportunistic microorganisms and a decrease in the amount of lactobacilli and bifidobacteria. L. casei IMV B-7280, B. animalis VKL and B. animalis VKB probiotic strains altered the microbiota spectrum of the vagina and intestine of Staphylococcus-infected mice: the amount of Lactobacillus and bifidobacteria increased with the reduction of the number of opportunistic microorganisms. Also under the influence of these strains, the normalization of the microbiota spectrum typical for vagina and intestine was observed in different periods of observation – in the intestines of mice the number of coliform bacteria increased, the number of microscopic fungi, streptococci and staphylococci decreased; in the vagina, the number of coliform bacteria and microscopic fungi decreased, the number of streptococci normalized. Rapid elimination of S. aureus 8325-4 from the vagina and prevention of the spread of infection to the intestine were observed after use of probiotics. Preventive effect of L. acidophilus IMV B-7279 and L. delbrueckii subsp. bulgaricus IMV B-7281 for bacterial vaginitis in mice was less effective. So, the target probiotic strains L. casei IMV B-7280, B. animalis VKL and B. animalis VKB are promising for the creation of highly effective novel probiotic drugs that can be used for directed prevention of infectious and inflammatory diseases of the genitourinary system caused by pathogenic and opportunistic microorganisms

Zonal pressure gradient along the equatorial Atlantic

For three consecutive periods during the summer of 1974, ships of many nations made observations along the Atlantic equator as part of the GATE program [GARP (Global Atmospheric Research Program) Atlantic Tropical Experiment]. Combining these observations, it is found that the zonal pressure gradient over the central Atlantic at the surface and at 50 dbar, relative to 500 dbar, increased from 3.2 to 7.3 and 2.2 to 5.3 × 10-5 dynes/g respectively between June/July and August and then held close to the high values in September...

Digital receivers for low-frequency radio telescopes UTR-2, URAN, GURT

This paper describes digital radio astronomical receivers used for decameter and meter wavelength observations. This paper describes digital radio astronomical receivers used for decameter and meter wavelength observations. Since 1998, digital receivers performing on-the-fly dynamic spectrum calculations or waveform data recording without data loss have been used at the UTR-2 radio telescope, the URAN VLBI system, and the GURT new generation radio telescope. Here we detail these receivers developed for operation in the strong interference environment that prevails in the decameter wavelength range. Data collected with these receivers allowed us to discover numerous radio astronomical objects and phenomena at low frequencies, a summary of which is also presented.Comment: 24 pages, 15 figure

Fast and reliable test for typing polymorphisms in genes tox3, slc4a7, map3k1 and fgfr2, connected with the risk of breast cancer development with pyrosequencing method

Данная статья посвящена разработке условий генотипирования полиморфизмов в генах TOX3, SLC4A7, MAP3K1 и FGFR2 методом пиросеквенирования и использования его для выявления предрасположенности к развитию cпорадического рака молочной железы у женщин. В результате проведенного исследования установлено повышение частоты встречаемости полиморфизма TT (Rs4973768) в гене SLC4А7 у больных раком молочной железы. Отношение шансов риска развития рака молочной железы для этого гена составляет 1,89 (ДИ = 1,01–3,397; р≤0,047), относительный риск 1,7 (р≤0,049). Отношение шансов для полиморфизма Rs3803662 составило 1,49, для Rs889312 — 1,59 и для Rs2981582 — 1,29. Предложенный метод анализа полиморфизмов в генах TOX3, SLC4A7, МAP3K1 и FGFR2 с помощью технологий пиросеквенирования позволяет быстро проводить анализ аллельных вариантов полиморфизмов и может быть использован при оценке риска развития спорадического рака молочной железы.Search for markers of classification of patients with high or low risk of disease development is the most important task of the molecular epidemiology. The prognostication of risk of some illnesses can be fulfilled with the help of classical risks factors, such as index of body weight, level of lipids, smoking, family history and genetic factors (polymorphism in genes BRCA1 and BRCA2 when talking about inherited breast carcinoma) etc. But these findings are not enough for the prognostication of risk of sporadic breast cancer. Occurrence of research with full genomic analysis of polymorphism on DNA microchips allowed to reveal more than 60 new polymorphisms, which can be potential markers of breast cancer development. Some of such allelic variants revealed during this research were polymorphisms in genes TOX3, SLC4A7, MAP3K1 and FGFR2

Superconducting Receivers for Space, Balloon, and Ground-Based Sub-Terahertz Radio Telescopes

We give a review of both our own original scientific results of the development of superconducting receivers for sub-terahertz astronomy and the main leading concepts of the global instrumentation. The analysis of current astronomical problems, the results of microwave astroclimate research, and the development of equipment for sub-terahertz radio astronomy studies justify the need and feasibility of a major infrastructure project in Russia to create a sub-terahertz telescope, as well as to enhance the implementation of the ongoing Millimetron and Suffa projects. The following results are discussed: i) superconducting coherent receivers and broadband subterahertz detectors for space, balloon, and ground-based radio telescopes have been developed and tested; ii) ultrasensitive receiving systems based on tunnel structures such as superconductor-insulator-superconductor (SIS) and superconductor-insulator-normal metal-insulator-superconductor (SINIS) have been created, fabricated, and examined; iii) a receiving array based on SINIS detectors and microwave readout system for such structures has been implemented; iv) methods for manufacturing high-quality tunnel structures Nb/AlOx/Nb and Nb/AlN/NbN based on niobium films with a current density of up to 30 kA/cm(2) have been developed. Receivers operated at 200 to 950 GHz and having a noise temperature only a factor of 2 to 5 higher than the quantum limit have been created and tested