Description of Transport Tunnel in Haloalkane Dehalogenase Variant LinB D147C+L177C from Sphingobium japonicum

The activity of enzymes with active sites buried inside their protein core highly depends on the efficient transport of substrates and products between the active site and the bulk solvent. The engineering of access tunnels in order to increase or decrease catalytic activity and specificity in a rational way is a challenging task. Here, we describe a combined experimental and computational approach to characterize the structural basis of altered activity in the haloalkane dehalogenase LinB D147C+L177C variant. While the overall protein fold is similar to the wild type enzyme and the other LinB variants, the access tunnels have been altered by introduced cysteines that were expected to form a disulfide bond. Surprisingly, the mutations have allowed several conformations of the amino acid chain in their vicinity, interfering with the structural analysis of the mutant by X-ray crystallography. The duration required for the growing of protein crystals changed from days to 1.5 years by introducing the substitutions. The haloalkane dehalogenase LinB D147C+L177C variant crystal structure was solved to 1.15 angstrom resolution, characterized and deposited to Protein Data Bank under PDB ID 6s06

Analysis of temperaturepain sensitivity in patients with consequences of the cervical spinal cord injury

Background. The standard neurological assessment in patients with long-term consequences of spine-and-spinal cord injury and severe neurological deficit does not allow to accurately identify changes in sensitivity that determine the level, degree and nature of spinal cord injury, as well as to evaluate the minimal dynamics of these disorders with different treatment options. As a result, an objective instrumental assessment of the sensory sphere in the long-term period of spinal cord injury has not lost its relevance.The aim. To conduct an instrumental study of the temperature-pain sensitivity condition in patients with partial gross damage to the cervical spinal cord in the long-term period of the disease (type B on the ASIA scale).Methods. We examined 23 patients with consequences of vertebral fractures of the cervical spine in the late period of traumatic spinal cord disease, Grade B on the ASIA scale ASIA. The clinical analysis of sensitive disorders was performed according to ISNCSCI and ASIA scales. While studying the temperature-pain sensitivity the threshold of thermal sensitivity and the threshold of pain from hot were determined in СIV–SI dermatomes on the right and on the left using an electricesthesiometer.Results. The examined patients had hypesthesia of heat and pain sensitivity, hyperesthesia of pain sensitivity, thermoanesthesia and thermoanalgesia. The degree of changes in the temperature-pain sensitivity depended on the topographic localization of dermatomes. The more distally the study area was located from the level of damage, the more pronounced the disorders were. In 30.4 % of patients, the pain sensitivity from hot in the chain of dermatomes from CIV to SI was preserved on at least one side. The combination of thermoanesthesia with thermoanalgesia was observed in 69.6 % of cases in dermatomes with ThVII and distally.Conclusions. The instrumentally registered level of the temperature-pain sensitivity disorder did not correspond to clinically determined localization of sensory disorders. The range of discrepancy ranged from 2 to 12 dermatomes, with defining the sensitivity subclinical deficit over the area of clinical sensory disorders

Role of active endoscopic tactic in patients with acutely bleeding gastric cancer

Departamentul de chirurgie nr. 3, Universitatea Naţională de Medicină “A.Bogomoletz”, Kiev, Ucraina, Al XII-lea Congres al Asociației Chirurgilor „Nicolae Anestiadi” din Republica Moldova cu participare internațională 23-25 septembrie 2015Introducere: Hemoragia masivă din cancerul gastric (CG) este rară, însă gravă, cu consecinţe primejdioase, ce creşte morbiditatea şi mortalitatea. Frecvenţa acestei complicaţii a CG creşte progresiv şi doar cîteva lucrări referitoare la aceas tă problemă au fost publicate pe durata ultimilor 30 de ani. Scopul studiului: evaluarea frecvenţei şi severităţii clinice a hemoragiei acute din cancerul gastric (HACG), cît şi studierea rolului endoscopiei de urgenţă în determinarea caracteristicilor HACG şi monitorizarea endoscopică pentru crearea unui algoritm de management chirurgical. Material şi metode: În perioada 1997-2014, în Centrul de Hemoragii Gastrointestinale (HGI) din Kiev au fost trataţi 1125 pacienţi cu HACG (vârsta medie: 56,6±9,2 ani). Analiza rezultatelor a fost realizată în două perioade. Perioada I (1997-2009) a reflectat analiza retrospectivă a 817 pacienţi trataţi cu HACG şi dezvoltarea unui algoritm de management chirurgical, cu accent pe rolul şi importanţa semnelor endoscopice ale HACG şi perioada II (2010-2014) – punerea în aplicare a algoritmului propus prin examen endoscopic la 308 pacienţi. Rezultate: Diagnosticul de CG era cunoscut anterior în 238 (21,2%) cazuri, în timp ce în 887 (78,8%) cazuri acesta a fost stabilit la internarea pacientului cu HGI. Gradul I de HACG a fost relevat la 46 (4,1%) pacienţi, II – 210 (18,7%), III – 402 (35,7%), IV – 467 (41,5%). Examenul endoscopic de urgenţă a relevat stigmatele FI – 11,5%, FII – 41,0%, FIII – 47,5%, iar hemostaza endoscopică repetată cu scop de prevenire a resîngerării a fost efectuată în cazul stigmatelor FI şi FII la 986 pacienţi (83,1%). Efectuarea operaţiei de urgenţă s-a redus de la 10,5% la 6,2%; în timp ce mortalitatea postoperatorie s-a micşorat de 2,7 ori. Concluzii: Algoritmul propus de management chirurgical ameliorează rezultatele tratamentului.Introduction: Major bleeding at gastric cancer (GC) are rare but is serious with dangerous effects, increased morbidity and mortality. Frequency of this complication GC progressively increases and only a few reports on such problem have been published over the last 30 yrs. The aim: Our study focuses on the evaluation of frequency, clinical severity of acute bleeding gastric cancer (ABGC) importance of the role emergency endoscopy in determining the features of ABGC and endoscopic monitoring for algorithm of surgical tactics. Material and methods: From 1997 to 2014, 1125 patients (the average age 56.6±9.2 years) with ABGC were treated at Kyiv Emergency Center of GIB. Analysis of results was carried out in two periods. I period (1997-2009) was retrospective analysis of 817 treated patients with ABGC and algorithm development for surgical tactics with an emphasis on the role and significance of endoscopic features of ABGC and II period (2010-2014) was implementation of proposed endoscopic algorithm in 308 patients. Results: The diagnosis GC was known before in 238 (21.2%) and established only after admission for AGIB in 887 (78.8%) pts. I stage ABGC had 46(4.1%), II – 210 (18.7%), III – 402 (35.7%), IV – 467 (41.5%) pts. Emergency endoscopy revealed stigmata FI – 11.5%, FII – 41.0%, FIII – 47.5%, repeated endoscopic methods for hemostasis prophylactics of rebleeding and monitoring have used for stigmata FI, FII (83.1%) in 986 cases. Emergency surgery was reduced from 10.5% to 6.2%, postoperative mortality in 2.7 times. Conclusions: The designed algorithm of surgical tactics improves results of treatment

Comparative analysis of dynamics in thermal pain sensitivity after correction of severe and mild spine deformities in patients with idiopathic scoliosis

Introduction Despite a great number of researches on idiopathic scoliosis reported there is still no instrumentation assessment of sensitivity before and after surgical correction of the curve found in literature. The purpose of the work was to explore dynamics in thermal pain sensitivity following correction of severe and mild spinal curves in patients with idiopathic scoliosis. Material and methods The work included results of examination of 25 patients with idiopathic scoliosis. Sampling population was divided into two groups depending on the extent of preoperative curve in major arch: Group I included patients with the curve of ≤ 60º (15 cases); Group II consisted of patients with the curve of > 60º (10 patients). Preoperative neurological examination showed no motor, reflex and sensory impairments. Acute deformity correction and spine stabilization with transpedicular systems were produced in all the cases. Thermal pain sensitivity was explored preoperatively and postoperatively at Th1-S2 dermatomes using electric esthesiometer. Results Disturbed thermal pain sensitivity of various extent was observed preoperatively in the study dermatomes of all patients with idiopathic scoliosis and was not shown to be dependent on the amount of the curve. Positive dynamics in thermal pain sensitivity revealed itself in reduced pain and heat thresholds and restored heat sensitivity and was observed in 41.5 to 54.1 % of the cases Group I following spine deformity correction. Deterioration was seen in 29.2 to 34.7 % of the cases. Group II showed negative dynamics in thermal pain sensitivity in 35.4 to 50 % of the cases with either increased or decreased pain threshold, enhanced heat threshold and loss of heat sensitivity. Sensitivity improved in 29.4 to 31.8 % of the patients. No changes in neurological status were noted in both groups after surgical correction with subclinical changes in thermal pain sensitivity to be considered. Conclusion Positive dynamics in thermal pain sensitivity prevailed in patients with idiopathic scoliosis following baseline curve correction of ≤ 60º. Negative dynamics in thermal pain sensitivity was seen in patients with baseline curve correction of > 60º with a greater risk of neurological complications

ЗАБОЛЕВАЕМОСТЬ И СМЕРТНОСТЬ ОТ РАКА ШЕЙКИ МАТКИ В ОМСКОЙ ОБЛАСТИ

The purpose of the study was a comparative assessment of the cervical cancer incidence and mortality among females in urban and rural populations of the Omsk region.Material and Methods. The study included females aged over 18 years from urban and rural areas of the Omsk region. According to population-based cancer registry data for the period 2004–16, the crude cancer incidence and mortality rates in urban and rural populations were calculated. The variational and correlation analyzes were used, the differences were assessed using Student t-test.Results. During 2004–16, the cervical cancer incidence among females of the Omsk region showed an increasing tendency, reaching a peak incidence in women aged 35–54. Concerning the cervical cancer mortality rate, it tended to decrease. For rural females, the cervical cancer mortality rate decreased from 10.8 to 9.7 per 100,000 females (p>0.05). For urban females, the mortality rate decreased from 9.2 to 7.5 per 100,000 females (p>0.05). No significant differences in the mortality rates between urban and rural females were found (p>0.05). The tendency towards decreased incidence of advanced cervical cancer (the average decrease being 3.3 % for rural females and 2.8 % for urban females, p>0.05) and increased incidence of early stage cervical cancer (the average rise being 1.8 % and 1.9 %, respectively) was observed.Conclusion. In the Omsk region, the cervical cancer mortality rate for both rural and urban females showed a tendency to decrease. During the study period, the incidence of cervical cancer had increased; however the incidence of advanced cervical cancer had decreased.Целью исследования явилась сравнительная характеристика динамики первичной заболеваемости смертности от рака шейки матки (РШМ) в г. Омске и сельских районах Омской области.Материал и методы. Объектом исследования было женское население Омска и муниципальных районов области в возрасте старше 18 лет. На основе массива данных канцер-регистра в 2004–16 гг. БУЗОО «КОД» рассчитаны интенсивные («грубые») показатели в зависимости от района проживания (город или село), проведено выравнивание временных рядов. Использованы вариационный и корреляционный виды анализов, оценка достоверности проведена с использованием t-критерия Стьюдента.Результаты. В целом по региону в течение анализируемого периода сформировалась тенденция к росту заболеваемости РШМ, большинство заболевших женщин относится к наиболее активной в трудовом и социальном плане возрастной группе – 35–54 года. За анализируемый период зарегистрировано значимое снижение летальности от РШМ: в г. Омске – в 2,6 раза; у жительниц сельских районов показатель снизился в 1,7 раза, достоверных различий между уровнями летальности в городе и сельской местности не выявлено (p>0,05). Положительным изменениям в динамике заболеваемости и смертности от РШМ способствовала сформировавшаяся тенденция к снижению числа случаев выявления запущенных стадий РШМ (средний темп убыли показателя в селе составил 3,3 %, в городе – 2,8 %, различия не достоверны (p>0,05)), а также незначительное увеличение удельного веса рака шейки матки ранней стадии (средний темп прироста – 1,8–1,9 %). Индекс накопления контингентов свидетельствует об уровне выживаемости больных с онкологической патологией. Кроме того, на величину этого показателя оказывают влияние количество впервые выявленных случаев РШМ и качество регистрации движения больных.Заключение. В Омской области сложилась благоприятная тенденция снижения смертности и летальности от РШМ как в городе, так и в сельских районах. За анализируемый период наблюдается рост заболеваемости РШМ, при этом снижается удельный вес случаев выявления заболевания в терминальной (IV) стадии

Клиническое наблюдение пациента с внебольничной пневмонией, вызванной Legionellа pneumophila

A clinical case of community-acquired pneumonia (CAP) caused by Legionellа pneumophila is described in the article. A 37-year-old male, a worker of a nuclear power station, was admitted with dry cough, dyspnea on light exertion, and fever. The disease onset was acute; the patient had been unsuccessfully treated with b-lactam antibiotics for 5 days before the admission. Two weeks before the disease onset, he had been to a neighbor region by business and stayed in a hotel. Non-severe bilateral CAP caused by Legionella was diagnosed using standard methods including Legionella rapid urine antigen test. Pneumonia was complicated by pleural exudate and moderate respiratory failure (class III according to PORT scale). Clinical particularities included diarrhea in the first day of the disease, elevated hepatic enzyme levels in blood, elevated blood urea, and hepatomegaly. Adequate specific therapy with levofloxacin and supplemental oxygen resulted in slow resolving of physical signs of pneumonia and pleuritis during 14 days with complete resolution of clinical symptoms and laboratory abnormalities and long-term maintenance of pulmonary infiltration.Представлено описание клинического случая внебольничной пневмонии (ВП), вызванной Legionellа pneumophila. Мужчина 37 лет, работник атомной электростанции, госпитализирован с жалобами на сухой кашель, одышку при незначительной физической нагрузке, лихорадку. Заболел остро, амбулаторно в течение 5 дней получал β-лактамные антибиотики – без эффекта; 2 нед. назад находился в соседнем регионе в командировке, где проживал в гостинице. При лабораторном и инструментальном обследовании, включавшем стандартные методы и экспресс-тест на наличие легионеллезной антигенурии, диагностирована двусторонняя легионеллезная ВП нетяжелого течения, осложнившаяся развитием экссудативного плеврита и дыхательной недостаточности II степени (III класс риска по шкале PORT). Особенности клинической картины: наличие у пациента в первые сутки заболевания диареи, повышение уровня аспартатаминотрансферазы, γ-глутамилтрансферазы, мочевины при лабораторном, гепатомегалия – при инструментальном исследовании. На фоне адекватной этиотропной терапии (левофлоксацин) и оксигенотерапии отмечалось медленное разрешение аускультативных и перкуторных изменений, обусловленных пневмонией и плевритом (14 дней) при практически полном регрессе клинических симптомов и лабораторных признаков, длительное сохранение пневмонической инфильтрации

Colonial Green Alga Botryococcus is a Producer of Valuable Metabolites

Впервые исследован изолят зеленой водоросли Botryococcus braunii, выделенный из озера Шира в 2022 году, как продуцент разрушаемых биопластиков полигидроксиалканоатов (ПГА) и углеводородов. Определен состав жирных кислот, углеводородов и полигидроксиалканоатов. Углеводороды, синтезируемые изолятом, представлены в основном диенами с длиной углеродной цепи С27 и С29. Показана неспособность аксеничной культуры B. braunii к синтезу ПГА и наличие в биомассе альгологически чистой и неаксеничной культуры полимера (до 7–10 %). Изучена бактериальная компонента консорциума «водоросль-бактерии», в котором обнаружены и идентифицированы бактерии Pseudomonas mendocina, Pseudomonas koreensis, Aeromonas hydrophila, способные к синтезу ПГА. Показано, что альгологически чистая и неаксеничная культура B. braunii является источником не только водорослевых углеводородов, но также и разрушаемых биопластиков ПГАAn isolate of the green alga Botryococcus braunii collected from Lake Shira in 2022 has been studied for the first time as a producer of degradable bioplastics polyhydroxyalkanoates (PHAs) and hydrocarbons. The compositions of fatty acids, hydrocarbons, and PHAs were determined. The hydrocarbons synthesized by the isolate were mainly represented by dienes with carbon chain lengths C27 and C29. The study demonstrated the inability of the axenic culture of B. braunii to synthesize PHAs and the presence of polymer in the biomass of unialgal and non-axenic culture (up to 7–10 %). The bacterial component of the algal- bacterial consortium was studied, and bacteria Pseudomonas mendocina, Pseudomonas koreensis, and Aeromonas hydrophila, capable of PHA synthesis, were discovered in it and identified. The study showed that the unialgal and non-axenic culture of B. braunii can be a source of not only algal hydrocarbons, but also degradable bioplastics PHA

Clinical observations of minimal gastrostomy through minilaparotomy in palliative patients

Gastrostomy is used to feed palliative patients with dysphagia. Currently, the preference is given to percutaneous puncture methods of gastrostomy, which do not require general anesthesia. Percutaneous puncture techniques are possible only if the patency of the upper parts of the digestive tract still exists for the «pull method» and can require additional X-ray irradiation in case of the «push method». These operations require expensive disposable kits, which affects the prevalence and availability of the technique. Therefore, the use of an alternative minimally invasive gastrostomy through minilaparotomy is justified. Minimal-invasive pressure gastrostomy is known for a long time, and in combination with small access, it can be successfully used to provide nutrition for palliative patients with dysphagia. Most patients with dysphagia have a thin anterior abdominal wall, which allows using minimal access. It is important to choose the right place of the incision so that access is in the area of the formation of the fistula. To obtain additional diagnostic information one can use a radiography of the abdominal cavity, which shows the gas bubble of the stomach, and other high-tech methods: ultrasound, spiral computed tomography, etc. At the same time such patients do not require general anesthesia, it is possible to perform the operation under a local anesthesia. The article provides a detailed description of the technique of minimal invasive laparotomy gastrostomy and two clinical observations of palliative patients who underwent this operation

Интернализация рекомбинантной имиглюцеразы в перитонеальные макрофаги мыши и фибробласты мыши линии L929

Enzyme replacement therapy (ERT) is one of the most efficient treatments for lysosomal storage diseases. Type 1 Gaucher disease is caused by β-glucocerebrosidase enzyme deficiency, which may be compensated for by intravenous infusions of imiglucerase—a recombinant enzyme. Imiglucerase targets macrophages and enters these cells via interaction with mannose receptors on the cell membrane. Characterisation of internalization of enzymes by target cells is important in the context of the development of new medicines and production of existing ERT medicines. The peritoneal and alveolar macrophages, as well as macrophages of the spleen of small laboratory animals (rats and mice) are widely used in such studies. However, isolation of cells from animal sources raises ethical issues, and therefore continuous mammalian cell lines may offer an attractive alternative. The aim of the study: to conduct comparative studies on the internalization of recombinant imiglucerase into mouse peritoneal macrophages and L929 mouse fibroblasts. Materials and methods: CerezymeR batches 7HV0913, C6214H05, 7HV0888 (Genzyme Ltd., UK); Glurazim batches 020416, 011117, 021117 (LLC “IBC “Generium”, Russia). We used peritoneal macrophages obtained from BALB/c mice and L929 mouse fibroblasts. The cells were cultured in DMEM/F12 complete growth medium with 10% fetal bovine serum. The activity of imiglucerase internalized into the cells was evaluated spectrophotometrically by hydrolysis of the artificial substrate—4-methylumbelliferyl-β-Dglucopyranoside. Results: the study compared internalization of recombinant imiglucerase (the active ingredient of CerezymeR and Glurazim) by mouse peritoneal macrophages and L929 mouse fibroblasts. It was demonstrated that the medicines activity in the lysates of peritoneal macrophages is comparable with that in the lysates of L929 mouse fibroblasts. Regardless of the model system, the activity of Glurazim stayed within the acceptable range (80–125%) established for biosimilar products. Conclusions: the experiments proved that L929 mouse fibroblasts could be recommended for assessment of internalization of recombinant imiglucerase.Фермент-заместительная терапия (ФЗТ) является одной из самых действенных при лечении болезней лизосомального накопления. Болезнь Гоше первого типа характеризуется недостатком нативного фермента β-глюкоцереброзидазы, который возмещают внутривенными инфузиями рекомбинантного фермента (имиглюцераза). Клетками-мишенями имиглюцеразы являются макрофаги, в которые фермент проникает посредством взаимодействия с рецепторами маннозы на клеточной мембране. Оценка интернализации ферментов клетками-мишенями представляет интерес при разработке новых и воспроизведении существующих препаратов для ФЗТ. Для этих исследований широко применяются перитонеальные и альвеолярные макрофаги, макрофаги селезенки мелких лабораторных животных (крыс и мышей). Однако получение таких клеток затрагивает этические вопросы использования лабораторных животных. Альтернативой являются перевиваемые клеточные линии млекопитающих. Цель работы: провести сравнительные исследования интернализации рекомбинантной имиглюцеразы в перитонеальные макрофаги мыши и фибробласты мыши линии L929. Материалы и методы: Церезим®, серии 7HV0913, C6214H05, 7HV0888 (Джензайм Лтд., Великобритания); Глуразим, серии 020416, 011117, 021117 (ООО «МБЦ «Генериум», Россия). В работе использовали перитонеальные макрофаги, полученные от мышей линии BALB/c, и фибробласты мыши линии L929. Клетки культивировали в полной ростовой среде ДМЕМ/Ф12 c добавлением 10% сыворотки плода крупного рогатого скота. Активность имиглюцеразы, проникшей в клетки, оценивали спектрофотометрически по гидролизу искусственного субстрата 4-метилумбеллиферил-β-D-глюкопиранозида. Результаты: представлены данные сравнительной оценки интернализации рекомбинантной имиглюцеразы, действующего вещества препаратов Церезим® и Глуразим, перитонеальными макрофагами мыши и клетками фибробластов мыши линии L929. Показано, что активность препаратов в лизатах перитонеальных макрофагов сопоставима с их активностью в лизатах клеток фибробластов мыши линии L929, при этом активность разработанного препарата Глуразим независимо от типа клеток, была в границах допустимого диапазона (80–125%), установленного для биоподобных препаратов. Выводы: экспериментально доказано, что фибробласты мыши линии L929 могут быть рекомендованы для оценки интернализации рекомбинантной имиглюцеразы