Construction of stably maintained non-mobilizable derivatives of RSF1010 lacking all known elements essential for mobilization

<p>Abstract</p> <p>Background</p> <p>RSF1010 is a well-studied broad-host-range plasmid able to be mobilized to different bacteria and plants. RSF1010-derived plasmid vectors are widely used in both basic research and industrial applications. In the latter case, exploiting of mobilizable plasmids or even the plasmids possessing negligible mobilization frequency, but containing DNA fragments that could promote conjugal transfer, is undesirable because of biosafety considerations. Previously, several mutations significantly decreasing efficiency of RSF1010 mobilization have been selected. Nevertheless, construction of the RSF1010 derivative lacking all known loci involved in the conjugal transfer has not been reported yet.</p> <p>Results</p> <p>Novel non-mobilizable derivatives of RSF1010 lacking all known DNA sequences involved in the mobilization process have been obtained due to the exploiting of λRed-driven recombination between the plasmid and a constructed <it>in vitro </it>linear DNA fragment. To provide auto-regulated transcription of the essential replication gene, <it>repB</it>, the plasmid loci <it>oriT</it>, <it>mobC </it>and <it>mobA </it>were substituted by the DNA fragment containing P_{<it>lac</it>UV5}→<it>lacI</it>. Mobilization of the obtained RSFmob plasmid was not detected in standard tests. The derivative of RSFmob with increased copy number has been obtained after <it>lacI </it>elimination. High stability of both constructed plasmids has been demonstrated in <it>Escherichia coli </it>and <it>Pantoea ananatis</it>. Design of RSFmob allows easy substitution of P_{<it>lac</it>UV5}by any desirable promoter for construction of novel derivatives with changed copy number or host range.</p> <p>Conclusion</p> <p>Novel non-mobilizable derivatives of RSF1010 lacking all known DNA sequences involved in the mobilization process and stably maintained at least in <it>E. coli </it>and <it>P. ananatis </it>have been constructed. The obtained plasmids became the progenitors of new cloning vectors answering all biosafety requirements of genetically modified organisms used in scale-up production.</p

Повышение психологической компетентности сестринского персонала в педиатрии: направления, формы, содержание

Relevance. In recent years the powers of nurses have been considerably expanded, which determines the increase of their responsibilities and the need of permanent improving of their skills.Object: determination of directions, forms and contents of training courses for increasing psychological competence of pediatric nurses in connection with their status and work experience.Methods and the object of research. The main method was a questionnaire developed by the article’s authors. 59 pediatric nurses from different regions of the Russian Federation who participated in remote educational cycles in Moscow and Yekaterinburg have been included in the survey. Nurses were divided into two groups based on their status and experience in the specialty. The group consisted of 31 respondents (average age 41,2 years). 28 respondents were Managers of nursing (average age 40,8 years).Results and conclusions. Stress management skills are more essential for nurses than for nursing managers and nursing staff with work experience of up to 10 years and over 20 years. Communication skills are relevant for nursing managers and nursing staff with experience of over 20 years. For all the nurses, situational assistance in complicated communicative situations and stress management is a priority compared to the long-term psychological support. The choice of education form (seminar, training) does not depend on the nursing staff’s status and work experience.Актуальность. В последние годы значительно расширяются полномочия медицинской сестры, что определяет уровень ее ответственности и необходимость постоянного повышения своей квалификации.Цель: определение направлений, форм и содержания образовательных курсов повышения психологической компетентности педиат рических медицинских сестер в связи со статусом и стажем работы. Методы и объект исследования. Основным методом являлась оригинальная анкета, разработанная авторами статьи. В анкетирование были включены педиат рические медицинские сестры (59 человек) из различных регионов Российской Федерации, которые участвовали в выездных образовательных циклах в Москве и Екатеринбурге в 2014 г. Медицинские сестры были разделены на две группы с учетом статуса и стажа работы по специальности. Группа медицинских сестер состояла из 31 респондента (средний возраст 41,2 года). Менеджеры сестринского дела представлены 28 респондентами (средний возраст 40,8 года).Результаты и выводы. Навыки управления стрессом востребованы в большей степени медицинскими сестрами, чем менеджерами сестринского дела, и сестринским персоналом со стажем до 10 и более 20 лет. Коммуникативные навыки актуальны для менеджеров сестринского дела и сестринского персонала со стажем более 20 лет. Ситуативная помощь по управлению сложными коммуникативными ситуациями и стрессом является приоритетной по сравнению с долгосрочным психологическим сопровождением для всех медицинских сестер. Выбор формы обучения (семинары, тренинги) не зависит от статуса и рабочего стажа сестринского персонала

Chronic endometritis and infertility — in vitro fertilization outcomes: systematic review and meta-analysis

Background. The relevance of the problem is related to the high prevalence of chronic endometritis (CE); its role in female infertility, implantation failures during assisted reproductive technology procedures, and recurrent miscarriage; as well as the lack of a unified strategy in the diagnosis and treatment of this pathology. The present systematic review with a meta-analysis focuses on evaluating the impact of CE and its therapy on the outcome of in vitro fertilization. In addition, the effect of CE of various severity on the outcomes of assisted reproductive technologies is analyzed. Objective. To analyze the effect of CE of varying severity and its treatment on the outcomes of in vitro fertilization. Methods. Using PubMed, Medline, Scopus, Embase, ELibrary, Cochrane Central Register of Controlled Trials (CENTRAL), WHO International Clinical Trials Registry, and Russian Science Citation Index, a systematic search was conducted for articles published over the past 12 years that met the following criteria: randomized controlled trial examining the effect of CE of varying severity on fertility and ways to treat it. The following indicators were calculated: ongoing pregnancy/live birth, clinical pregnancy, and miscarriage rates. A total of 4145 patients (from ten studies) were included. A meta-analysis was performed using Stata 11.0 software (The Cochrane Collaboration, Oxford, UK). The heterogeneity was considered low at I2 &lt;30%, moderate at 30–50%, and high at &gt;50%. Results. Women with CE exhibited lower ongoing pregnancy/live birth (OR 1.97; p = 0.02) and clinical pregnancy rates (OR 2.28; p = 0.002) as compared to women without it. CE treatment increased the ongoing pregnancy/live birth (OR 5.33; p &lt; 0.0001) and clinical pregnancy rates (OR 3.64; p = 0.0001). In vitro fertilization outcomes were comparable in women treated for CE and women without CE (ongoing pregnancy/live birth rate, clinical pregnancy rate, and miscarriage rate: p = ns). Women with severe CE exhibited lower ongoing pregnancy/live birth (OR 0.43; p = 0.003) and clinical pregnancy rates (OR 0.40; p = 0.0007). Mild CE showed no significant effect on in vitro fertilization outcomes (ongoing pregnancy/ live birth rate, clinical pregnancy rate, and miscarriage rate: p = ns). Conclusion. The conducted meta-analysis showed that CE significantly reduces the ongoing pregnancy/live birth and clinical pregnancy rates in infertile women undergoing in vitro fertilization. Noteworthy is that antimicrobial therapy in such patients improves the results of assisted reproductive technologies, which are comparable to those of patients without CE. The negative impact of this pathology on the implantation capacity of the endometrium is most often observed in the severe form, while its mild form has virtually no effect on the in vitro fertilization outcome

Yarrowia lipolytica growth under increased air pressure: influence on enzymes production

Improvement of microbial cell cultures oxygenation can be achieved by the increase of total air pressure, which increases oxygen solubility in the medium. In this work, a pressurized bioreactor was used for Yarrowia lipolytica batch cultivation under increased air pressure from 1 to 6 bar. Cell growth was strongly enhanced by the pressure rise. Fivefold and 3.4-fold increases in the biomass production and in specific growth rate, respectively, were observed under 6 bar. The increase of oxygen availability caused the induction of the antioxidant enzyme superoxide dismutase, which indicates that the defensive mechanisms of the cells against oxidative stress were effective and cells could cope with increased pressure. The pregrowth of Y. lipolytica under increased pressure conditions did not affect the lipase production ability of the cells. Moreover, the extracellular lipase activity increased 96% using a 5-bar air pressure instead of air at 1- bar pressure during the enzyme production phase. Thus, air pressure increase in bioreactors is an effective mean of cell mass and enzyme productivity enhancement in bioprocess based in Y. lipolytica cultures

Comparison of Yarrowia lipolytica and Pichia pastoris cellular response to different agents of oxidative stress

Yeast cells exposed to adverse conditions employ a number of defense mechanisms in order to respond effectively to the stress effects of reactive oxygen species. In this work, the cellular response of Yarrowia lipolytica and Pichia pastoris to the exposure to the ROSinducing agents’ paraquat, hydrogen peroxide, and increased air pressure was analyzed. Yeast cells at exponential phase were exposed for 3 h to 1 mM paraquat, to 50 mM H2O2, or to increased air pressure of 3 or 5 bar. For both strains, the cellular viability loss and lipid peroxidation was lower for the cells exposed to increased air pressure than for those exposed to chemical oxidants. The glutathione induction occurred only in Y. lipolytica strain and reached the highest level as a response to PQ exposure. In general, antioxidant enzymes were more expressed in Y. lipolytica than in P. pastoris. The enzyme superoxide dismutase was induced in both strains under all the oxidant conditions but was dependent on the cellular growth phase, being undetectable in non-growing cells, whereas glutathione reductase was more induced in those conditions. Hydrogen peroxide was the most efficient inducer of catalase. Both yeast cultures underwent no cellular growth inhibition with increased air pressure, indicating that these yeast species were able to adapt to the oxidative stressful environment.The authors acknowledge the financial support provided by "Fundacao para a Ciencia e Tecnologia" (Grant SFRH/BD/47371/2008)

Russian clinical practice guidelines «congenital adrenal hyperplasia»

Congenital adrenal hyperplasia (CAH) is a group of autosomal recessive diseases characterized by a defect in one of the enzymes or transport proteins involved in the cortisol synthesis in the adrenal cortex. The most common form of CAH, which occurs in more than 90% of cases, is a 21-hydroxylase enzyme deficiency. The latter is subdivided into nonclassical and classic (salt-losing and virilizing) forms. The prevalence of classic forms of 21-hydroxylase deficiency ranges from 1: 14,000 to 1:18,000 live births worldwide. According to the data of neonatal screening in the Russian Federation, the prevalence of the disease in some regions ranges from 1: 5000 to 1: 12000, in the country as a whole - 1: 9638 live newborns. The non-classical form of CAH occurs more often - from 1: 500 to 1: 1000 among the general population. In second place is the hypertensive form of CAH - a deficiency of 11β-hydroxylase, which, according to the literature, occurs in about 1 per 100,000 newborns. These clinical guidelines were compiled by a professional community of narrow specialists, approved by the expert council of the Ministry of Health of the Russian Federation, and updated the previous version published in 2016. The clinical guidelines are based on systematic reviews, meta-analyses and original articles, and scientific work on this issue in the Russian Federation and other countries. The purpose of this document is to provide clinicians with the most up-to-date, evidence-based guidelines for the CAH diagnosis and treatmen

Kinematic analysis of the daily activity of drinking from a glass in a population with cervical spinal cord injury

Background Three-dimensional kinematic analysis equipment is a valuable instrument for studying the execution of movement during functional activities of the upper limbs. The aim of this study was to analyze the kinematic differences in the execution of a daily activity such as drinking from a glass between two groups of patients with tetraplegia and a control group. Methods A total of 24 people were separated into three groups for analysis: 8 subjects with metameric level C6 tetraplegia, 8 subjects with metameric level C7 tetraplegia and 8 control subjects (CG). A set of active markers that emit infrared light were positioned on the upper limb. Two scanning units were used to record the sessions. The activity of drinking from a glass was broken down into a series of clearly identifiable phases to facilitate analysis. Movement times, velocities, and the joint angles of the shoulder, elbow and wrist in the three spatial planes were the variables analyzed. Results The most relevant differences between the three groups were in the wrist. Wrist palmar flexion during the back transport phase was greater in the patients with C6 and C7 tetraplegia than in the CG, whereas the highest wrist dorsal flexion values were in forward transport in the subjects with C6 or C7 tetraplegia, who required complete activation of the tenodesis effect to complete grasping. Conclusions A detailed description was made of the three-dimensional kinematic analysis of the task of drinking from a glass in healthy subjects and in two groups of patients with tetraplegia. This was a useful application of kinematic analysis of upper limb movement in a clinical setting. Better knowledge of the execution of this movement in each of these groups allows therapeutic recommendations to be specifically adapted to the functional deficit present. This information can be useful in designing wearable robots to compensate the performance of AVD, such as drinking, in people with cervical SCI