The swine intestinal wall thickening dynamic in result of different pressure applying inside the welding anastomotic instrument model

Introduction. The life-time studies of intestine operate at a pressure of 0.003-0.006 (N/m2). To create magnetic ring anastomosis 0.1-0.4 N/m2 pressure is used. The right initial compression of intestine walls before radio frequency electric influence isn’t established properly, but obviously has to homogenize the tissues dense. Several publications according 1.125 N/mm2 pressure was found. Aim to investigate the thinning dynamics of intestinal walls having human-sized dimensions during the high pressure action, in attempt to determine the optimal moment of anastomotic welding beginning. Material and methods. The swine organ complex was delivered to laboratory during 6 hours at 4 оС, and then was heated to 28-32 оС into 0.9% NaCl. Two intestine walls having human sizes were positioned on electrodes inside the anastomotic device prototype. After fixation, tissues were pressed for 60 or 120 seconds. The 30 experiments were provided using pressure values 2.1, 3.0, 3.9 and 5.0 N/mm2 for tissues thinning dynamic investigation by connected to electrodes micrometer device. Results and discussion. The thinning of two intestinal walls, occurs as result of applied by electrodes external pressure, valued in range from 2.1 or 3.9 N/mm2, significantly reduces it’s tempo from 35th to 60th action sec. We allow that this resistance derives from the mucous and submucosal layers. Under a pressure of 1.1 N/mm2, the similar thickness degree and tempo is achieved by pressure prolongation to 120 sec, but under 5.0 N/mm2 it needs 20 sec. The pressure by 3.0-3.9 N/mm2 over 35 sec, or 5.0 N/mm2 over 20 sec are resulted in much lower tissue stabilizing level. Obviously, its structural ground is a resistance combination of all intestinal layers at the crash edge. Conclusions. By choosing the appropriate pressure value from 1.1 to 5.0 N/mm2 and its exposure time from 20 to 120 seconds, it is possible to adjust the degree of tissue dense before the subsequent connection

Особливості та ефективність застосування електрозварювання живих тканин у виконанні хірургічного втручання та досягненні локального контролю при лікуванні місцево поширеного раку ободової кишки

The aim of the work: to determine the peculiarities and efficiency of the live tissue electric welding technology applying for surgical proceeding and local control achieving during the advanced colon cancer treatment. Materials and Methods. In 57 patients, welding mobilization and/or anastomosis was performed during surgical interventions, radical or palliative. Inclusion criteria were adenocarcinoma, T3-4, N+, it’s abscessing. The electric welding clinical devices EK-300M1 and Patonmed EKVZ-300 (Ukraine) were used, equipped with special instruments. The features of the resection line, healing and anastomotic function were researched, and the appearance of local tumor recurrence was observed up to 10 years after the surgery. The average patients age was (68.3±8.4) years. Results and Discussion. Electro-welded intestinal anastomoses were made by multi-point (7) and one-stage (8) methods, sutured (42) – by single stitches. Electric welding mobilization took place along the narrow line of current passing, in the planned by surgeon tissues, had not affected by the tumor, regardless of their density and inflammatory changes, at dry operation field because of simultaneous welding hemostasis. The sealing of the lymph nodes capsule and simultaneous closing of lymphatic vessels were occurred. Right orienting of electric current by a surgeon facilitated plane mobilization of specimen. Along with the division, the coagulation pseudocapsule was formed along separated tissue edges. It was possible to mobilize the tumor and infected tissues in case of decay, without violation of the abscess walls integrity. The detection and skeletonizing of organ vessels for their central overlap, which was closed by welding, without imposing a ligature, was provided. Characteristics of electric welding resulted in a slight exudation by drainage, which gave the possibility of reliable control of leakage and anastomotic tightness. Any leakage of electric welding anastomosis was observed, but in sutured one leakage was found in 2 (4.8 %) as grade ІІІ, in 6 (14.3%) grade І or ІІ. The stool, respectively, was observed through (1.9±0.3) days, and (2.6±0.3) days. For 2 years there were observed 12 patients having electric weld anastomosis, for 3 years – 8.5 years – 3 patients, 10 years – 1 patient. Having suture anastomosis were observed for 2 years 34 patients, for 3 years – 20.5 years – 9 patients, 10 years – 3 patients. According to the endoscopic and radiological studies, the line of electro-welded colon connection was difficult to identify. The threads migration from sutured anastomosis line was observed up to 6 months. None of the observable patients in either groups revealed clinical, endoscopic or tomography signs of local tumor recurrence: external deformity of anastomosis or new tumor growth at the site of its removal. The electric welding usage provides an opportunity to achieve tumor removal within R0, in volume D2 and CME. Coagulation of the lymph nodes capsule and their vessels contributes to ablasty during lymphadenectomy. Coagulation pseudocapsule, beside oncological reason, could provide a reduction of postoperative complications number, even in the case of abscess removing or anastomotic dehicience, that might be an important survival background. The vascular closage line properties, made by welding, provides the secondary bleeding prevention in such case.Цель работы: определить особенности и эффективность применения технологии электросварки живых тканей в выполнении хирургического вмешательства и достижении локального контроля при лечении распространенного рака ободочной кишки. Материалы и методы. В 57 больных осуществляли электросварочную мобилизацию и/или наложение анастомоза во время оперативных вмешательств, радикальных или паллиативных. Критериями включения были аденокарцинома, T3-4, N+, абсцедирование. Использовали источники электросварочных импульсов ЕК-300М1 и “Патонмед” ЕКВЗ-300 (Украина), укомплектованные инструментами. Исследовали особенности линии резекции, заживления и функцию анастомоза, наблюдали появление локального рецидива опухоли до 10 лет после вмешательства. Средний возраст больных 68,3±8,4 лет. Результаты исследований и их обсуждение. Электросварной кишечный анастомоз накладывали многоточечным (7) и одномоментным (8) методами, шовный (42) – одиночными стежками. Электросварная мобилизация происходила вдоль узкой линии прохождения тока, в намеченных пределах не пораженных опухолью тканей, независимо от их плотности и воспалительных изменений, на фоне обеспечения гемостаза. Происходило уплотнение лимфатических узлов и одновременное перекрытие лимфатических сосудов. Ориентировка хирургом электросварного тока облегчало футлярную мобилизацию. Одновременно с разделением, вдоль разделенных краев ткани образовывалась коагуляционная псевдокапсула. Без нарушения целостности стенок абсцесса удавалось мобилизовать опухоль и инфицированные ткани в случае распада. Обеспечивалось выделение и скелетизация магистральных питающих сосудов для их центрального перекрытия, которое осуществляли без наложения лигатуры. Особенности электросварки обусловили незначительную выраженность экссудации по дренажам, обеспечивало возможность надежного контроля герметичности анастомоза. Несостоятельность электросварного анастомоза не наблюдали, шовного наблюдали у 2 больных (4,8 %) – III степени, у 6 (14,3 %) I–II степени. Наблюдали в отдаленном периоде с электросварным анастомозом в течение 2 лет – 12 больных, 3 лет – 8, 5 лет – 3, 10 лет – 1. С шовным анастомозом в течение 2 лет наблюдали 34 больных, 3 лет – 20, 5 лет – 9, 10 лет – 3. По данным эндоскопического и рентгенологического исследования, линию электросварного соединения кишки было трудно идентифицировать, а из линии шовного до 6 месяцев наблюдали миграцию нитей. Ни у одного из наблюдаемых больных обеих групп не обнаружили клинических, эндоскопических или томографических признаков местного рецидива опухоли: внешней деформации анастомоза или нового роста опухоли в месте ее удаления. Применение электросварки позволяет достичь удаления опухоли в пределах R0, в объеме D2 и СМЕ. Коагуляция капсулы лимфатических узлов и их сосудов способствует абластичности лимфаденэктомии. Коагуляционная псевдокапсула может обеспечить уменьшение количества послеоперационных осложнений даже в случае удаления абсцесса или несостоятельности швов анастомоза, что может быть важной предпосылкой выживаемости. Свойства электросварного перекрытия сосудов способствуют избежанию вторичного кровотечения в такой ситуации.Мета роботи: визначити особливості та ефективність застосування технології електрозварювання живих тканин у виконанні хірургічного втручання та досягненні локального контролю при лікуванні поширеного раку ободової кишки. Матеріали і методи. У 57 хворих здійснювали електрозварну мобілізацію та/або накладання анастомозу під час операційних втручань, радикальних або паліативних. Критеріями включення були аденокарцинома, T3-4, N+, абсцедування. Використовували джерела електрозварювальних імпульсів ЕК-300М1 та “Патонмед” ЕКВЗ-300 (Україна), укомплектовані інструментами. Досліджували особливості лінії резекції, загоєння і функцію анастомозу, та спостерігали появу локального рецидиву пухлини до 10 років після втручання. Результати досліджень та їх обговорення. Електрозварний кишковий анастомоз накладали багатоточковим (7) та одномоментним (8) методами, шовний (42) – поодинокими стібками. Електрозварна мобілізація відбувалась вздовж вузької лінії проходження струму, у визначених межах не уражених пухлиною тканин, незалежно від їх щільності та запальних змін на тлі забезпечення гемостазу. Відбувалось ущільнення лімфатичних вузлів та одночасне перекриття лімфатичних судин. Орієнтування хірургом електрозварного струму полегшувало футлярну мобілізацію. Одночасно з розділенням, вздовж розділених країв тканини утворювалась коагуляційна псевдокапсула.         Без порушення цілісності стінок гнійника вдавалось мобілізувати пухлину та інфіковані тканини в разі розпаду. Забезпечувалось виявлення та скелетизація магістральних живлячих судин для їх центрального перекриття, яке здійснювали без накладання лігатури. Особливості електрозварювання зумовлювали незначну вираженість ексудації по дренажах, що надавало можливість надійного контролю герметичності анастомозу. Неспроможність електрозварного анастомозу не спостерігали, шовного виявили у 2 хворих (4,8 %) – ІІІ ступеня, у 6 хворих (14,3 %) І–ІІ ступеня. Спостережено у віддаленому періоді з електрозварним анастомозом протягом 2 років – 12 хворих, 3 років – 8, 5 років – 3, 10 років – 1. З шовним анастомозом протягом 2 років спостережено 34 хворих, 3 років – 20, 5 років – 9, 10 років - 3. За даними ендоскопічного та рентгенологічного дослідження лінію електрозварного з’єднання кишки було складно ідентифікувати, а з лінії шовного до 6 місяців спостерігали міграцію ниток. У жодного зі спостережених хворих обох груп не виявили клінічних, ендоскопічних чи томографічних ознак місцевого рецидиву пухлини: зовнішньої деформації анастомозу або нового росту пухлини в місці її видалення. Застосування електрозварювання надає можливість досягти видалення пухлини в межах R0, в обсязі D2 та СМЕ. Коагуляція капсули лімфатичних вузлів та їх судин сприяє абластичності лімфаденектомії. Коагуляційна псевдокапсула може забезпечити зменшення кількості післяопераційних ускладнень навіть у разі видалення абсцесу або неспроможності швів анастомозу, що може бути важливою передумовою виживання. Властивості електрозварного перекриття судин сприяють запобіганню вторинній кровотечі в такій ситуації

Surgical treatment of wounded persons with complicated thoracic damages

Objective. To improve the results of complex surgical treatment of wounded and injured persons with thoracic trauma, complicated by bronchopleural fistulas, pleural empyema and defects in the thoracic wall soft tissues, basing on introduction of the proposed procedure of vacuum therapy in combination with bronchoscopic obturation of certain bronchus. Materials and methods. Analysis of results of the complex surgical help delivery for 54 wounded persons with defects of the thoracic wall soft tissues, broncho-pleural fistulas and pleural empyema, caused by the combat thoracic trauma, on ІІІ and ІV levels of medical support while conduction of the Antiterrorist operation/Operation of the Joined Forces (the main group) and 73 injured persons with thoracic trauma of the peace period (the comparison group) in the 2014 - 2019 yrs period was conducted. Results. Improvement of complex surgical treatment of the wounded and damaged persons with thoracic trauma and defects of the thoracic wall soft tissues have permitted to reduce the complications frequency by 6.7%, and lethality - by 9.2% in the main group. Conclusion. There was proposed a procedure of vacuum therapy, ultrasound cavitation together with bronchoscopic bronchial obturation for its fistula, which have proved its efficacy in complex surgical treatment of severely wounded persons with the gun-shot thoracic damages and presence of combination of pleural empyema, bronchial fistula and defects of soft tissues

Nucleologenesis in the Caenorhabditis elegans Embryo

In the Caenorhabditis elegans nematode, the oocyte nucleolus disappears prior to fertilization. We have now investigated the re-formation of the nucleolus in the early embryo of this model organism by immunostaining for fibrillarin and DAO-5, a putative NOLC1/Nopp140 homolog involved in ribosome assembly. We find that labeled nucleoli first appear in somatic cells at around the 8-cell stage, at a time when transcription of the embryonic genome begins. Quantitative analysis of radial positioning showed the nucleolus to be localized at the nuclear periphery in a majority of early embryonic nuclei. At the ultrastructural level, the embryonic nucleolus appears to be composed of a relatively homogenous core surrounded by a crescent-shaped granular structure. Prior to embryonic genome activation, fibrillarin and DAO-5 staining is seen in numerous small nucleoplasmic foci. This staining pattern persists in the germline up to the ∼100-cell stage, until the P4 germ cell divides to give rise to the Z2/Z3 primordial germ cells and embryonic transcription is activated in this lineage. In the ncl-1 mutant, which is characterized by increased transcription of rDNA, DAO-5-labeled nucleoli are already present at the 2-cell stage. Our results suggest a link between the activation of transcription and the initial formation of nucleoli in the C. elegans embryo

Distinct Expression/Function of Potassium and Chloride Channels Contributes to the Diverse Volume Regulation in Cortical Astrocytes of GFAP/EGFP Mice

Recently, we have identified two astrocytic subpopulations in the cortex of GFAP-EGFP mice, in which the astrocytes are visualized by the enhanced green–fluorescent protein (EGFP) under the control of the human glial fibrillary acidic protein (GFAP) promotor. These astrocytic subpopulations, termed high response- (HR-) and low response- (LR-) astrocytes, differed in the extent of their swelling during oxygen-glucose deprivation (OGD). In the present study we focused on identifying the ion channels or transporters that might underlie the different capabilities of these two astrocytic subpopulations to regulate their volume during OGD. Using three-dimensional confocal morphometry, which enables quantification of the total astrocytic volume, the effects of selected inhibitors of K+ and Cl− channels/transporters or glutamate transporters on astrocyte volume changes were determined during 20 minute-OGD in situ. The inhibition of volume regulated anion channels (VRACs) and two-pore domain potassium channels (K2P) highlighted their distinct contributions to volume regulation in HR-/LR-astrocytes. While the inhibition of VRACs or K2P channels revealed their contribution to the swelling of HR-astrocytes, in LR-astrocytes they were both involved in anion/K+ effluxes. Additionally, the inhibition of Na+-K+-Cl− co-transporters in HR-astrocytes led to a reduction of cell swelling, but it had no effect on LR-astrocyte volume. Moreover, employing real-time single-cell quantitative polymerase chain reaction (PCR), we characterized the expression profiles of EGFP-positive astrocytes with a focus on those ion channels and transporters participating in astrocyte swelling and volume regulation. The PCR data revealed the existence of two astrocytic subpopulations markedly differing in their gene expression levels for inwardly rectifying K+ channels (Kir4.1), K2P channels (TREK-1 and TWIK-1) and Cl− channels (ClC2). Thus, we propose that the diverse volume changes displayed by cortical astrocytes during OGD mainly result from their distinct expression patterns of ClC2 and K2P channels

Simulation-based cheminformatic analysis of organelle-targeted molecules: lysosomotropic monobasic amines

Cell-based molecular transport simulations are being developed to facilitate exploratory cheminformatic analysis of virtual libraries of small drug-like molecules. For this purpose, mathematical models of single cells are built from equations capturing the transport of small molecules across membranes. In turn, physicochemical properties of small molecules can be used as input to simulate intracellular drug distribution, through time. Here, with mathematical equations and biological parameters adjusted so as to mimic a leukocyte in the blood, simulations were performed to analyze steady state, relative accumulation of small molecules in lysosomes, mitochondria, and cytosol of this target cell, in the presence of a homogenous extracellular drug concentration. Similarly, with equations and parameters set to mimic an intestinal epithelial cell, simulations were also performed to analyze steady state, relative distribution and transcellular permeability in this non-target cell, in the presence of an apical-to-basolateral concentration gradient. With a test set of ninety-nine monobasic amines gathered from the scientific literature, simulation results helped analyze relationships between the chemical diversity of these molecules and their intracellular distributions

European Red List of Habitats Part 1. Marine habitats

The European Red List of Habitats provides an overview of the risk of collapse (degree of endangerment) of marine, terrestrial and freshwater habitats in the European Union (EU28) and adjacent regions (EU28+), based on a consistent set of categories and criteria, and detailed data and expert knowledge from involved countries1. A total of 257 benthic marine habitat types were assessed. In total, 19% (EU28) and 18% (EU28+) of the evaluated habitats were assessed as threatened in categories Critically Endangered, Endangered and Vulnerable. An additional 12% were Near Threatened in the EU28 and 11% in the EU28+. These figures are approximately doubled if Data Deficient habitats are excluded. The percentage of threatened habitat types differs across the regional seas. The highest proportion of threatened habitats in the EU28 was found in the Mediterranean Sea (32%), followed by the North-East Atlantic (23%), the Black Sea (13%) and then the Baltic Sea (8%). There was a similar pattern in the EU28+. The most frequently cited pressures and threats were similar across the four regional seas: pollution (eutrophication), biological resource use other than agriculture or forestry (mainly fishing but also aquaculture), natural system modifications (e.g. dredging and sea defence works), urbanisation and climate change. Even for habitats where the assessment outcome was Data Deficient, the Red List assessment process has resulted in the compilation of a substantial body of useful information to support the conservation of marine habitats

Involvment of Cytosolic and Mitochondrial GSK-3β in Mitochondrial Dysfunction and Neuronal Cell Death of MPTP/MPP+-Treated Neurons

Aberrant mitochondrial function appears to play a central role in dopaminergic neuronal loss in Parkinson's disease (PD). 1-methyl-4-phenylpyridinium iodide (MPP+), the active metabolite of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), is a selective inhibitor of mitochondrial complex I and is widely used in rodent and cell models to elicit neurochemical alterations associated with PD. Recent findings suggest that Glycogen Synthase Kinase-3β (GSK-3β), a critical activator of neuronal apoptosis, is involved in the dopaminergic cell death. In this study, the role of GSK-3β in modulating MPP+-induced mitochondrial dysfunction and neuronal death was examined in vivo, and in two neuronal cell models namely primary cultured and immortalized neurons. In both cell models, MPTP/MPP+ treatment caused cell death associated with time- and concentration-dependent activation of GSK-3β, evidenced by the increased level of the active form of the kinase, i.e. GSK-3β phosphorylated at tyrosine 216 residue. Using immunocytochemistry and subcellular fractionation techniques, we showed that GSK-3β partially localized within mitochondria in both neuronal cell models. Moreover, MPP+ treatment induced a significant decrease of the specific phospho-Tyr216-GSK-3β labeling in mitochondria concomitantly with an increase into the cytosol. Using two distinct fluorescent probes, we showed that MPP+ induced cell death through the depolarization of mitochondrial membrane potential. Inhibition of GSK-3β activity using well-characterized inhibitors, LiCl and kenpaullone, and RNA interference, prevented MPP+-induced cell death by blocking mitochondrial membrane potential changes and subsequent caspase-9 and -3 activation. These results indicate that GSK-3β is a critical mediator of MPTP/MPP+-induced neurotoxicity through its ability to regulate mitochondrial functions. Inhibition of GSK-3β activity might provide protection against mitochondrial stress-induced cell death