Determination of the Critical Phases of the Experimental Research Using Laboratory Animals: Risk Analysis

Critical phases (stages) of preclinical studies are those procedures or types of research activities whose accurate and correct implementation is a prerequisite for obtaining valid and reliable results. Russian and foreign standards require determination of the critical phases of each individual study by quality assurance staff based on checking the study protocol (plan).The aim of the study was to identify critical phases typical for most preclinical studies, and assess the potential risks during inspections.Materials and methods: the study was carried out by analysing the types and consequences of nonconformities. Numerical parameters of risks were analysed for each critical phase of the preclinical study identified by quality officers of the Joint Stock Company “Scientific and Production Association ‘HOME OF PHARMACY’”.Results: it was discovered that incorrect implementation of a procedure constituted a potential nonconformity at all the identified critical phases, and a potential consequence was acquisition of low-quality data. A combination of incorrectly implemented procedures at two or more critical phases could pose an unacceptable risk and lead to complete loss of data or failure to process data, and, as a result, the need to repeat the study.Conclusions: the highest risk was identified for such critical phases as preparation and administration of final doses of test samples, performance of physiological tests, collection of biological material samples, and handling of biological material samples by other relevant departments. Summarising the data obtained on the risks of all the critical phases, it can be concluded that risk action should take the form of regular inspections by the quality assurance staff and the study director. By adjusting the frequency of inspections, the risk of each critical phase can be made insignificant

A novel-type luciferin from Siberian luminous earthworm Fridericia heliota : structure elucidation by spectral studies and total synthesis

Author Posting. © The Author(s), 2014. This is the author's version of the work. It is posted here by permission of Wiley-VCH Verlag GmbH & Co for personal use, not for redistribution. The definitive version was published in Angewandte Chemie International Edition 53 (2014): 5566–5568, doi:10.1002/anie.201400529.We report structure elucidation and synthesis of the luciferin from the recently discovered luminous earthworm Fridericia heliota. This luciferin represents a key component of a novel ATP-dependent bioluminescence system. The UV, fluorescence, NMR and HRMS spectral studies were performed on 5 mkg of the isolated substance, and gave four isomeric structures, conforming with spectral data. These isomers were chemically synthesized and one of them was found to produce light in the reaction with a protein extract from Fridericia. The novel luciferin was found to have an unusual deeply modified peptidic nature, implying an unprecedented mechanism of action.We acknowledge support from the Program of the Government of the Russian Federation “Measures to attract leading scientists to Russian educational institutions” (grant no. 11. G34.31.0058), the programs MCB RAS, President of the Russian Federation “Leading science school” (grant 3951.2012.4) and the Russian Foundation for Basic Research (grant 14-03-01015). B.M.S. was supported by a stipend from the Program of the President of the Russian Federation.2015-04-1

ДОСВІД ПІДГОТОВКИ СТУДЕНТІВ ДО УЧАСТІ У ВСЕУКРАЇНСЬКІЙ СТУДЕНТСЬКІЙ ОЛІМПІАДІ З КЛІНІЧНОЇ ІМУНОЛОГІЇ ТА АЛЕРГОЛОГІЇ (підсумки викладачів та погляд студентів)

The work adduces a generalization experience in carrying out the I stage of the student’s competition in Clinical Immunology and Allergology within training of students for participation in the II stage of the All-Ukrainian student’s competition. The aim of this work was to show possible ways of increasing the educational motivation of students and quality of education in this clinical cycle. During 3 years students of 5th, 6th courses take part in the I stage of the All-Ukrainian student’s competition, that is held at the Department of Internal Medicine № 2 and Occupational Diseases, of State Institution “Dnipropetrovsk Medical Academy of the Ministry of Health of Ukraine”. Stages, which are part of the the intra high school competition program, are described.The conclusions contain recommendations: carrying out professional competitions gives an ability to sum up the results of work on teaching the discipline “Clinical Immunology and Allergology”; competitions contribute to the goals and main task of each university and teacher – to improve quality of preparing qualified specialists;  carrying out the I stage and preparation to the second stage of the All-Ukrainian student’s competition is desirable to held with involvement of leading experts – the practicing doctors, giving opportunities to students to gain not only profound theoretical knowledge, but also practical experience that in general promotes self–affirmation of students and increase of scientific and practical potential of the students.У роботі представлено узагальнення досвіду з проведення І етапу студентської олімпіади з клінічної імунології та алергології в рамках підготовки студентів до участі в ІІ етапі Всеукраїнської студентської олімпіади.  Метою проведеної роботи було показати можливі шляхи підвищення навчальної мотивації студентів та якості навчання на клінічному циклі. На кафедрі внутрішньої медицини № 2 та профпатології  ДЗ “Дніпропетровська медична академія МОЗ України” протягом 3-х років проводиться І етап Всеукраїнської студентської олімпіади, в якій беруть участь студенти 5, 6 курсів. Описано етапи, які входять  до програми внутрішньовузівської олімпіади. У висновках надаються рекомендації: проведення фахової олімпіади надає змогу підвести підсумки роботи з викладання дисципліни “Клінічна імунологія та алергологія”; змагання сприяють виконанню мети і головного завдання вузу і кожного викладача – підвищення якості підготовки кваліфікованих спеціалістів; проведення І етапу та підготовку до ІІ етапу Всеукраїнської студентської олімпіади бажано здійснювати із залученням провідних фахівців - практичних лікарів, що надає змогу студентам отримати не тільки глибокі теоретичні знання, але й  практичний досвід, що загалом сприяє самоствердженню студентів та підвищенню науково-практичного потенціалу студентського корпусу

New records of lichens and allied fungi from the Leningrad Region, Russia. XI

Twelve lichen species and two lichenicolous fungi, of them seven that belong to Micarea prasina group, are reported for the first time for St. Petersburg or the whole Leningrad Region. The lichenicolous fungus Intralichen baccisporus is new to Russia, and the lichen Micarea nowakii – for European Russia. A comparative table of characteristics for seven species of Micarea prasina group is presented

Features of Chronic Bronchitis in Different Age Groups

Background: Lung diseases are assuming greater relevance and importance today. Chronic bronchitis is a self-nosology, which may precede the development of COPD, the importance of which can hardly be overestimated. The main problem in this disease is caused by late diagnosis and treatment due to the delay by patients in seeking medical help. The aim of the work was to study the distribution and exposure to tobacco smoke, especially chronic bronchitis, depending on various factors, including age. Methods: We examined 1779 persons, including 855 men and 924 women. The mean age of the population was 35.83±8.3 years. We conducted surveys and spirometry. The outcome was assessed after a bronchodilation test was performed with salbutamol 400 mcg. We performed all statistical analysis using software package Statistica 10. Results: We identified chronic bronchitis in 9.2% of the cases in the group of younger individuals and in 14.9% of the cases in the group of older individuals, during the active detection of chronic bronchitis using questionnaires. The prevalence of cigarette smoking was slightly higher among the younger (39.5%) than the older persons (33.6%); the frequency of smoking in a group of chronic bronchitis was reliably higher. Also, in this group, the performance spirometry reliably decreased. Conclusions: Outpatient survey is an effective method of identifying chronic bronchitis. Smoking is a major risk factor in the group of young respondents and the prevalence of smoking is inversely related to the education level of the respondents, regardless of age. As the decline in the Forced Expiratory Volume (FEV1 and FEV1/FVC) is the main criterion diagnosis of COPD, it revealed significant declines in the FEV1 of the younger smoking individuals, which may help to predict the development of COPD in the older age group

Hormonal type in the conditions of experimental multiple skeletal trauma

In experiment reproduced model of multiple skeletal trauma (fracture of bones of right forearm and lift shin) with stabilization of fragments with external fixation devices in 35 Chinchilla rabbits. All manipulations were realized, at general anesthesia. Standard treatment included anesthesia, antibiotic prophyliaxis and. infusion therapy by common method. Decrease of metabolism activity was registered in the first group of laboratory animals at the standard treatment. That's why besided strandard treatment dexamethasone and adrenaline in average recommended doses was prescripted in the second group of rabbits in postoperative period for correction of posttraumatic hypobiosis. Laboratory control in several hours after the injection confirmed stable increase of level of corticosteroids in blood but it wasn't possible to register increase of concentration of adrenaline. Correlation analysis was realize to study the connection between the contentration of endogenous hormones in blood and. metabolism activity in rabbits in conditions of multiple skeletal trauma. At the consideration data as one array we determined high-significant (р < 0,001) direct moderate correlation between the values of rectal temperature and. the concentration of free thyroxine in blood (rs = 0,414), this connection was strong on the 3rd day after trauma (rs = 0,833; р < 0,001). Also indirect moderate correlation between values of rectal temperature and. the concentration of cortisol in blood of laboratory animals was registered (rs = 0,49; р < 0,001). The results of study of influence of adrenaline and. noradrenaline on the indices of metabolism activity weren't statistically significant. Obtained data allowed to make a conclusion about importance of endocrine activity of thyroid gland in the formation of adaptation strategy

Energy metabolism at experimental polytrauma: changes at the prescription of adrenaline and dexamethasone

The aim of the research was to study influence of dexamethasone and adrenaline on the concentration of energy metabolism substrates in blood of Chinchilla rabbits under the conditions of multiple skeletal trauma. We performed stabilization of the bones of right forearm and left shin in 35 animals by wire external fixation device with further osteotomy of bones of corresponding segments. We observed the decrease of activity of metabolism in rabbits on the background of standard treatment. Maintenance of normal activity of metabolism by prescription of adrenaline and dexamethasone caused more intensive use of energy resources of an organism. Imitation of hyperergic reaction of stress-realizing systems in rabbits caused the increase of concentration of glucose in blood and the decrease of concentration of triglycerides. High consumption of energy substrates was provided by more expressed catabolic reaction. The result of hypercatabolism was development of hypoproteinemia, hypoalbuminemia and increase of the risk of fluid shift from bloodstream into the tissues with formation of organ insufficiency

Molecular Genetic Studies of Intraoperative Ligamentum Flavum Bioptates of Patients with Spinal Canal Stenosis

Background. Ligamentum flavum is involved in the development of stenosis of the spinal canal and dural sac, but the mechanisms underlying degenerative-dystrophic changes have not yet been sufficiently studied. Aim. To study the expression of candidate genes potentially involved in connective tissue metabolism in intraoperative samples of Ligamentum flavum of patients with stenosis processes of spinal canal and duralsac on the lumbar spine.Materials and methods. During the surgical treatment, Ligamentum flavum biopsies were collected from 33 people with stenosis processes of spinal canal and dural sac on the lumbar spine (16 men, 17 women). RNA was isolated, then reverse transcription was performed. After that, real time PCR was performed (CFX96, Biorad) with the specific primers (RealTimePrimers.com).Results. The characteristics of the expression of candidate genes that are active in the Ligamentum flavum of  the examined patient population are given: NAT1 and NAT2 acetyltransferase genes; genes that determine the intensity of local metabolism (Dio1, Dio2 and Dio3); estradiol receptor genes ESR1 and ESR2; genes encoding receptors for growth factors and parathyroid hormone. All Ligamentum flavum samples showed activity of the NAT, ESR 2, FGFR1, FGFR3, NA20, PTH1R, and PTH2R genes. In the vast majority of samples (93.9–97.0 %), the following genes were active: PDGFA, ESR1, CALCR, and PDGFB. In 21.2–39.4 % of samples, “silence” of the Dio1, Dio2, Dio3, NAT2 and GDF5 genes was detected, and only 39.4 % of samples revealed NAT1 transcripts. The highest heterogeneity of transcript content was observed for Dio1, NAT2, and Dio2. The most stable expression in Ligamentum flavum tissue and a narrow range of fluctuations were characterized by genes with a high level of activity AANAT, ESR2, NAA20 and genes with an average level of activity FGFR1, FGFR3, PTH1R and PTH2R.Conclusions. Ligamentum flavum is a promising little-studied substrate for molecular genetic research. The revealed features of gene expression provide new information about pathogenesis, and the new fundamental knowledge obtained can provide a basis for developing methods for preventing the development of epidural fibrosis