Zone Of Necrosis Research Articles

Targeted Cx43 therapeutics reduce NLRP3 inflammasome activation in rat burn injury.

Burns are dynamic injuries characterized by an initial zone of necrosis that progresses to compromise surrounding tissue. Acute inflammation and cell death are two main factors contributing to burn progression. These processes are modulated by Connexin43 (Cx43) hemichannels and gap junctions in burns and chronic wounds. Particularly, Cx43-mediated ATP release may interact with the P2X7 receptor to activate the NLRP3 inflammasome pathway. This study used a deep partial thickness rat burn model to evaluate the effect of Cx43 antisense oligodeoxynucleotides (Cx43asODN) or the Cx43 hemichannel blocker Tonabersat for the inhibition of inflammasome activation and their use as potential treatments for burn injury. Using immunofluorescence analysis, our data showed that Cx43asODN or Tonabersat reduced Cx43 hemichannel and gap junction expression. Concomitantly, they marginally and transiently reduced P2X7 expression and inflammasome complex assembly and inflammation. Quantitative analysis using H&E, Masson's trichrome & Picrosirus Red revealed reduced epidermal thickness and improved collagen preservation in treated burn wounds. Collectively, our findings suggest a possible involvement of the Cx43-mediated NLRP3 inflammasome pathway via P2X7 activation in early burn wound healing. This indicates that targeting Cx43 may have a potential therapeutic effect to improve healing outcomes.

Mesh Refinement Compensation via Regression in thermal necrosis FEM

Abstract Finite Element Models (FEM) for simulating various physical phenomena are indispensable in today’s research. They are also applied in high-frequency (HF) surgical applications to simulate Joule heating and thermal necrosis formation in biological tissue. One challenge lies in choosing an appropriate mesh size, balancing between sufficiently accurate simulation results and a coarse mesh for fast computations. In this contribution, we investigate, using three mesh sizes (coarse, medium, and fine), the extent to which regression analysis of discrete simulation results of necrosis dimensions can compensate for mesh refinement in an FEM for HF soft coagulation. We conducted exponential regression analysis on discrete simulation results of the necrosis zone. It has been found that while it is generally possible, if the FEM simulation results already deviate significantly from the true solution, regression analysis cannot compensate for errors arising from the numerical method. Therefore, a coarse mesh with subsequent regression analysis can be utilized to determine necrosis propagation in depth, with a maximum error of only -3.4 % compared to the fine mesh within the considered interval. However, for the necrosis in the lateral direction, the maximum error is 44.3 %, which is too large. Even the mid-size mesh is not accurate enough in the lateral direction, necessitating the use of a fine mesh for the lateral direction.

Changes in ultrasonic elastometry parameters of the liver parenchyma during its radiofrequency ablation (experimental study)

Objective. To determine in the experiment the changes in the elasticity of the liver parenchyma during its radiofrequency ablation at different distances from the electrode and their correspondence to the zones of irreversible thermal damage of the tissue. Materials and methods. The elasticity of the parenchyma of six samples of isolated porcine liver during radiofrequency ablation in automatic mode for 12 min with an initial applicator power of 50 W and its subsequent automatic increase by 10 W/min until critical impedance values were reached was evaluated by ultrasonic elastometry with the determination of the Young's modulus. The elasticity of the liver in kilopascals was determined before the start of radiofrequency ablation, during its implementation every minute for 12 minutes and 15, 30 and 60 minutes after the procedure in three zones located at a distance of 1.0, 1.8 and 3.0 cm from the applicator. Results. Before radiofrequency ablation, the elasticity of the liver parenchyma ranged from 4.1 to 9.3 kPa and averaged (6.64 ± 1.55) kPa. At the maximum power of the applicator – (109.67 ± 4.97) W – the transverse size of the hyperechogenic “cloud” at the 12th minute of the procedure was (18.0 ± 1.41) mm. The value of Young's modulus in the first zone of elastometry statistically significantly increased from the 1st minute of radiofrequency ablation and by the 11th minute reached the level of (46.38 ± 5.43) kPa and did not change significantly thereafter. In the second zone, a statistically significant increase in the value of Young's modulus to (44.22 ± 6.55) kPa was observed throughout the procedure, and after its termination it changed statistically insignificantly. In the third zone, changes in the value of Young's modulus occurred 3 minutes after the start of the procedure and continued until its completion, but its maximum value – (15.63 ± 1.57) kPa – exceeded the baseline level only about 2 times, and an hour after the completion of radiofrequency ablation, the value of Young's modulus decreased statistically significantly. Conclusions. The stiffness of the parenchyma of isolated porcine liver increases significantly during radiofrequency ablation under conditions of its sufficient duration, and depending on the distance to the electrode, these changes have different phase character. In loci corresponding to the zone of irreversible tissue necrosis, the initial slow approximately twofold increase in Young's modulus during the first 3 to 4 minutes is followed by a rapid exponential increase in the next 5 to 6 minutes and the formation of a plateau with 6 to 8 times the initial level, after which the index does not change significantly. To determine the edge of liver parenchyma ablation by elastometry, in addition to the absolute value of Young's modulus at the end of radiofrequency ablation and the multiplicity of its increase relative to the baseline value, such criteria as the three–phase nature of the increase in this indicator and the absence of its decrease within an hour after the procedure are equally important.

Outcomes of Proximal Hybrid Arterial Reconstruction in Combination with Simultaneous Amputation in Dry Atherosclerotic Gangrene of Toes

INTRODUCTION: When treating atherosclerotic gangrene of the lower limb (LL), the surgeon faces the questions about the reasonability of vascular reconstruction and the optimal timing of amputation after surgery on the LL arteries. The answer to these questions is given by assessing the state of the microvasculature of the operated limb. With sufficient development of the microvasculature and good collateral circulation, it is possible to perform a simultaneous amputation after proximal reconstruction. In this situation, a clear demarcation of the zone of necrosis and reversible ischemia is required, which can be realized by the method of ultraviolet luminescence spectroscopy. AIM: To analyze the results of hybrid reconstructions on the LL arteries with multilevel diffuse atherosclerotic lesions and dry gangrene of toes (DGT). MATERIALS AND METHODS: A prospective, controlled, non-randomized study included 29 patients suffering from critical ischemia of the lower limbs and having DGT, who were operated on in the amount of hybrid arterial reconstruction. The patients were divided into two groups: patients of group 1 (n = 14) underwent restoration of the main blood flow at the level of the iliofemoral arterial segment using a hybrid method, with simultaneous minor amputation of LL at various levels; patients of group 2 (control group, n = 15) underwent a simultaneous proximal and distal hybrid operation, providing main blood flow through at least one of the lower leg arteries, followed by a minor amputation of the lower leg at various levels over the next 4–5 days. RESULTS: There were no statistically significant differences in the groups in the degree of decrease in luminescence intensity after vascular surgery. A histological examination of intraoperative preparations of DGT revealed necrosis of the cellular microenvironment at luminescence amplitude (1.0 ± 0.05) × 105 photons at 410 nm frequency. At luminescence amplitude not exceeding this level, signs of necrobiosis were noted. Luminescence level of ≥ 1.0 × 105 photons was used as the amputation boundary. In the case of an uncomplicated vascular stage of the operation, a comparable decrease in the conventional amputation boundary was noted in the study groups. In the early postoperative period, in patients of group 1, the level of inflammation markers, average number of bed-days, and the number of thrombotic complications were lower than in the control group (p 0.05). A strong correlation was recorded between the morphological signs of the acute phase of inflammation and the intensity of chemiluminescence (r = 0.7, p 0.005). CONCLUSION: In patients with DGT, at a luminescence amplitude on the lower leg and foot not exceeding 1.0 × 105 photons at 410 nm frequency and 0.7 × 105 photons at 450 nm frequency, an effective treatment method is restoration of the main blood flow in the iliofemoral segment using a hybrid method with simultaneous minor amputation at various levels of the foot. This luminescence level is the conventional boundary between necrotic changes and reversible ischemia (necrobiosis) of the soft tissues of the LL.

Temperature Simulation of an Ablation Needle for the Prediction of Tissue Necrosis during Liver Ablation

Background/Objectives: Microwave ablation (MWA) is the leading therapy method for treating patients with liver cancer. MWA simulation is used to further improve the therapy and to help develop new devices. Methods: A water-cooled ablation needle was reconstructed. MWA simulations of a polyacrylamide phantom were carried out and compared with a representative clinical example (tumor diameter: 8.75 mm). The Arrhenius damage model and a critical temperature approach of 60 °C were applied to assess the necrosis zones. Finally, the simulation results were compared to the corresponding MR measurements. Results: Most of the heating in the simulation took place at a distance of 5 mm along the transverse axis and 20 mm along the longitudinal axis above the needle tip. The calculated Dice scores for the Arrhenius model were 0.77/0.53 for the phantom/clinical case. For the critical temperature approach, Dice scores of 0.60/0.66 for the phantom/clinical case were achieved. Conclusions: The comparison between simulated and measured temperature increases showed an excellent agreement. However, differences in the predicted necrosis volume might be caused by omitting consideration of the heat sink effect, especially in the clinical case. Nevertheless, this workflow enables short MWA simulation times (approximately 3 min) and demonstrates a step towards possible integration into daily clinical use.

In Vivo Histological Study Evaluating Non-Ablative Fractional 1940-nm Laser.

Non-ablative fractional lasers (NAFL) have increased in demand compared to ablative laser treatments as they provide lesser down time, fewer side-effects, and are safer to use. Non-ablative fractional treatment with lasers ranging from 1320 to 1927-nm have been shown to be safe and effective for skin resurfacing procedures. The objective of this study is to investigate healing of the 1940-nm NAFL-induced microthermal treatment zones (MTZs) in human skin from a histologic perspective. Three subjects received 1940-nm NAFL treatment to test areas on the abdomen at various timepoints during the study. The minimum 5 mJ/MTZ and maximum 20 mJ/MTZ energy settings were used at 20% coverage. Biopsies were taken coinciding with immediately posttreatment, 1, 3, 7 days, and 6 weeks posttreatment. Blinded analysis of hematoxylin and eosin stained slides was performed to measure the width and depth of the MTZs and evaluate the inflammatory and healing response of the skin over time (immediately to 6 weeks posttreatment). Safety was evaluated by assessing local skin responses and adverse events immediately after treatment and at all study visits. Histological analysis of tissue following NAFL 1940-nm treatments showed mild early inflammatory response (presence of lymphotic infiltrate) in some test areas and zones of necrosis and coagulation having widths and depths (immediately-3 days posttreatment) that scaled with the 1940-nm pulse energy. Signs of healing such as presence of dermal mucin, evidence of fibrosis, and absence of necrosis were observed long-term (7 days to 6 weeks posttreatment). Evidence of the MTZ persisted beyond the 6-week study and was predicted to last for 100 days. All local clinical skin responses healed within 6 weeks and were limited to mild, transient erythema and edema which resolved in less than 12-24 h following treatment. No serious adverse events occurred during the study. NAFL 1940-nm treatments are safe for inducing small fractional coagulation and necrosis zones in abdominal skin. NAFL 1940-nm laser creates fractional columns of injury with sufficient depth and coverage that suggest effective skin resurfacing, like other non-ablative fractional lasers.

ПЕРФУЗИОННАЯ КОМПЬЮТЕРНАЯ ТОМОГРАФИЯ – НОВЫЕ ГОРИЗОНТЫ В ДИАГНОСТИКЕ ЗАБОЛЕВАНИЙ ПОДЖЕЛУДОЧНОЙ ЖЕЛЕЗЫ

Abstract. Introduction. According to the World Health Organization, the most common disease of the pancreas is acute pancreatitis, which can lead to the development of pancreatic necrosis. In this regard, early diagnosis and assessment of the pancreatic necrosis risk development are of paramount importance for choosing adequate treatment approaches and improving outcomes for patients. Radiological research methods have become the “gold” standard in clinical practice for the primary diagnosis of pancreatic diseases. Contrast-enhanced computed tomography has traditionally been used to visualize the pancreas and assess the extent of inflammation and necrotic changes. However, standard computed tomography has limitations in assessing tissue perfusion, which is an important aspect of the pathophysiology of acute pancreatitis. Perfusion computed tomography of the pancreas has become a colossal breakthrough in radiation imaging, has become widespread as a method that allows the quantitative assessment of blood supply to the pancreas, including information about microcirculation, and it can be used for the early detection of ischemic changes that precede tissue necrosis in the pancreas. Aim. To study the capabilities of perfusion computed tomography and computed tomography angiography of the pancreas for evaluating the area of pancreatic necrosis in acute pancreatitis, with the aim of improving diagnosis and planning treatment measures to minimize the consequences and increase patient survival. Materials and Methods. A single-center examination was carried out on 42 patients (18 women, 24 men, mean age 38±2 years) with suspected acute pancreatitis. All patients underwent perfusion computed tomography within the first 3 days; over the next 3-5 days, computed tomography angiography of the pancreas was performed with bolus intravenous contrast to confirm the development of necrotic changes. Results and Discussion. After performing perfusion computed tomography within the first 3 days from the onset of symptoms, areas of ischemic damage to the pancreas tissue were identified in 16 (38.1%) out of 42 patients. After computed tomography angiography, the development of necrotic changes was confirmed in 7 patients (16.7%). The localization of the necrosis zone detected in computed tomography coincided with the ischemic zones on the color hemodynamic maps of perfusion computed tomography.

Changes in Microwave Ablation Zone Dimensions after Transarterial Embolization in an Ex Vivo Human Kidney Perfusion Model

PurposeTo utilize a novel ex vivo perfused human renal model and quantify microwave ablation (MWA) size differences in renal tissue when combining MWA with transarterial embolization (TAE). Materials and MethodsHuman kidneys (n = 5) declined for transplantation were obtained and connected to a fluoroscopy-compatible ex vivo perfusion system. Two ablations—1 standard MWA and 1 TAE-MWA—were performed in each kidney for 2 minutes at 100 W using a MWA system (Solero Angiodynamics). MWA alone was performed in the upper pole. In the lower pole, MWA was performed after TAE with 40–90 μm radiopaque microspheres to achieve angiographic stasis. Ablation zones of coagulative necrosis were sectioned along the long axis and segmented for maximal short-axis diameter (SAD) and long-axis diameter (LAD) measurements. ResultsA total of 10 ablations (5 MWAs and 5 TAE-MWAs) were performed in 5 human kidneys. TAE-MWA resulted in significantly increased SAD, LAD, volume, and sphericity compared with standard MWA ± SD, with mean measurements as follows (5 standard MWAs ± SD vs 5 TAE-MWAs ± SD, 2-tailed t-test): (a) SAD, 1.8 cm (SD ± 0.1) versus 2.5 cm (SD ± 0.1) (P < .001); (b) LAD, 2.9 cm (SD ± 0.3) versus 3.2 cm (SD ± 0.1) (P = .039); (c) volume, 5.0 mL (SD ± 0.5) versus 11.0 mL (SD ± 0.7) (P < .001); and (d) sphericity, 0.4 (SD ± 0.2) versus 0.6 (SD ± 0.1) (P = .049). Histology demonstrated no differences in TAE-MWA other than concentrated microspheres. ConclusionsThis ex vivo human kidney perfusion model confirmed that combined MWA-TAE significantly increased ablation size and spherical shape compared with MWA alone.

Comparative assessment of structural changes in the prostate after bipolar plasma and thulium fiber laser enucleation of benign prostatic hyperplasia

Background. Benign prostatic hyperplasia (BPH) is a common disease in older men. Transurethral surgery in BPH is the gold standard for treatment, but the techniques differ in the energy used. Considering the different mechanisms of action of bipolar plasma and thulium fiber laser energy on prostate tissue, the study of structural changes under their influence is relevant.The aim. To study the features of structural changes in the prostate during bipolar plasma and thulium fiber laser enucleation of benign prostatic hyperplasia.Methods. Thirty one patients with BPH were divided into two groups. Group 1 (n = 17) underwent transurethral plasma enucleation of the prostate; Group 2 (n = 14) underwent transurethral thulium fiber laser enucleation. Fragments of the surgical prostate capsule were collected intraoperatively and were processed according to standard examination protocols using light and electron microscopy. The width of coagulation necrosis, the relief of the dissection line, the severity of cell and the prostate intercellular matrix destruction were taken into account.Results. Studies have shown the clinical and functional perioperative homology of the compared methods of surgical treatment of BPH. Data from light and electron prostate microscopy showed a greater damaging effect of bipolar plasma energy, which is manifested by a larger width of the zone of coagulation necrosis, a torn and raised appearance of the dissection line, and pronounced integrity violations of cellular elements and intercellular matrix components. Laser exposure causes less pronounced changes, which indicates a more gentle effect of the thulium fiber laser on the prostate components.Conclusion. The obtained microscopy results indicate that the intraoperative action of a thulium fiber laser is less traumatic for the cells and intercellular matrix of the surgical prostate capsule compared to bipolar plasma exposure.

Thermal Field Monitoring When Exposing Soft Tissues to Low Temperatures: Thermography Prospects and Limitations

The review analyzes the existing tools for monitoring the dynamics of thermal fi elds when exposing the soft tissues to low temperatures. Features of contact and non-contact temperature measurements have been considered, their capabilities and limitations have been noted. There was substantiated the need to develop the procedures of intra-operative temperature control. Special attention has been paid to the non-contact non-invasive infrared thermography. This method has been shown to be applied for intra-operative monitoring of the movement of the ice lump edge on the surface of tissues, detection of a disordered thermal symmetry of the ice spot, thermal fi eld dynamics on the surface of tissues inside and outside the area of the operative zone. However, thermal imaging control of the dynamics of the primary necrosis zone and the ice ball edge in the volume of tissues is possible only under certain parameters of cryoimpact, for example, with a short-term cooling of tissues with a quasi-point nitrogen cryoapplicator. The possibility of using thermography at other stages of cryosurgery is also considered, i. e. as the method of additional diagnosis at the stage of surgery planning, as well as during the post-surgery period to control healing, scarring, etc.

Experimental study of the interstitial effect of a semiconductor laser with a wavelength of 445 nm on a biological model

Introduction. This study presents the results of interstitial exposure of a semiconductor laser with a wavelength of 445 nm to biological tissue samples at different pulsed wave power in a constant mode, with a contact method.Aim. To study the interstitial effect of a semiconductor laser with a wavelength of 445 nm on experimental tissue samples in a constant mode at different power.Materials and methods. As an experimental sample, we used biological tissue with a developed vascular structure in the form of pig liver. The source of laser radiation was a semiconductor laser with a wavelength of 445 nm, with a power range from 0.5 to 4 watts. When working with biological tissue samples, we evaluated their external and internal changes after laser exposure. The exposure time during interstitial exposure was 1 mm/sec with a 20 mm immersion depth of the laser fiber into the fabric. The results of the macro and microscopic picture were evaluated using histological examination and morphometry of the zones of destruction and coagulation necrosis, on a transverse section of the tissue.Results. The results of an experimental study indicate that interstitial laser exposure has a pronounced coagulation effect combined with a cutting effect. The optimal combination of coagulation and cutting effect of exposure, accompanied by visual contractility of the tissue, without excessive carbonation at a power of 3.0 watts.Conclusion. The use of interstitial exposure of a semiconductor laser with a wavelength of 445 nm on experimental tissue samples in a constant mode at different power showed the predominance of the coagulation effect in combination with the cutting effect with a pronounced reduction in tissue volume. Experiments have shown that the power of 3 W is the optimal power of laser exposure in the interstitial method, in which there is a pronounced reduction in the volume of the studied drug without excessive carbonation.

Advanced technique of myocardial no-reflow quantification using indocyanine green.

The post-ischemic no-reflow phenomenon after primary percutaneous coronary intervention (PCI) is observed in more than half of subjects and is defined as the absence or marked slowing of distal coronary blood flow despite removal of the arterial occlusion. To visualize no-reflow in experimental studies, the fluorescent dye thioflavin S (ThS) is often used, which allows for the estimation of the size of microvascular obstruction by staining the endothelial lining of vessels. Based on the ability of indocyanine green (ICG) to be retained in tissues with increased vascular permeability, we proposed the possibility of using it to assess not only the severity of microvascular obstruction but also the degree of vascular permeability in the zone of myocardial infarction. The aim of our study was to investigate the possibility of using ICG to visualize no-reflow zones after ischemia-reperfusion injury of rat myocardium. Using dual ICG and ThS staining and the FLUM multispectral fluorescence organoscope, we recorded ICG and ThS fluorescence within the zone of myocardial necrosis, identifying ICG-negative zones whose size correlated with the size of the no-reflow zones detected by ThS. It is also shown that the contrast change between the no-reflow zone and nonischemic myocardium reflects the severity of blood stasis, indicating that ICG-negative zones are no-reflow zones. The described method can be an addition or alternative to the traditional method of measuring the size of no-reflow zones in the experiment.

Effects of Parallel Contact Cooling on Pulsed-Type, Bipolar Radiofrequency-Induced Tissue Reactions in an in vivo Porcine Model.

Skin cooling during laser or radiofrequency (RF) treatments is a method to minimize thermal damage to the epidermis, reduce pain, and decrease post-treatment downtime. We evaluated the effect of parallel contact cooling (PCC) on RF-induced thermal reactions in minipig skin in vivo after bipolar microneedling RF treatment. RF treatments were administered at frequencies of 0.5, 1, and 2 MHz with single (500 ms), six (1000 ms), and ten (5000 ms) sub-pulse packs to minipig skin with or without PCC. Subsequently, thermometric imaging and histology were used to analyze skin reactions to RF. Thermometric images showed that PCC promptly lowered skin temperature in the RF-treated area, with this effect persisting for over 60s. Regardless of the PCC, RF treatments lasting for 500 ms with a single pulse pack resulted in peri-electrode coagulative necrosis (PECN) zones and inter-electrode non-necrotic thermal reaction (IENT) zones in the dermis. In contrast, treatment lasting 5000 ms with 10 sub-pulse packs produced distinct IENT without notable PECN over a wide dermal area. Skin specimens obtained at 1 h and 3, 7, and 14 days after PCC-assisted RF treatments showed a higher degree of thermal tissue reactions in the deeper dermal regions compared to those after RF treatments without PCC. PCC-assisted RF treatment, utilizing an invasive bipolar microneedling device, enhanced RF-induced skin reactions in the mid to deep dermis while preserving the epidermis and upper papillary dermis from excessive thermal tissue injury.

Case report: Typical Classic Epithelioid Granulomatous Enteritis in one-humped camel (Camelus dromedarius) in Aswan Slaughterhouses, Egypt

Abstract: A Granulomas can appear in a variety of pathological situations. So, recognizing the granulomatous pattern is essential. because it is confined to a limited number of disorders, some of which are life-threatening. There is little pathological data on the cells that contribute to the formation of granulomas in camels. This study identified cells associated with granulomatous disorder based on histological findings. The intestinal tissues were brought to the Pathology Department, Faculty of Veterinary Medicine, Aswan University, with intestinal lesions for histological evaluation. One case of epithelioid granuloma in the small intestine of dromedary camels (Camelus dromedarius) was detected by histopathological examination. There is a central zone of necrosis. Necrotic material is amorphous, structureless, eosinophilic, and granular debris with lost cellular features. surrounded by epithelioid cells and connective tissue, which contains lymphocytes, macrophages, and plasma cells. Epithelioid cells have a polygonal, faintly granular cytoplasm. Epithelioid cells have ill-defined cell borders and elongated or ovoid vesicular central nuclei.

Model of toxic fibrosis in Wistar rats: morphological and molecular-genetic parameters of the transition point to cirrhosis

BACKGROUND: To date, when studying the cellular and molecular genetic mechanisms of liver fibrogenesis in experimental rat models, no attention is paid to the transition point of fibrosis to cirrhosis as a separate stage. As this pathology develops, changes in the cell phenotype and gene expression are dynamic in nature, so it is necessary to evaluate them in a long-term temporal dynamics.&#x0D; AIM: The aim of this work is to study the morphological and molecular genetic changes in the liver of Wistar rats during nodular parenchymal rearrangement.&#x0D; METHODS: Fibrosis and cirrhosis of the liver in male Wistar rats was induced with a freshly prepared solution of thioacetamide, which was administered intragastrically through a tube at a dose of 200 mg/kg of body weight 2 times a week for 13 weeks. The level of mRNA of the tweak (tnfsf12), fn14 (tnfrsf12a), ang, vegfa, cxcl12 (sdf), and mmp-9 genes in the liver was detected by real-time polymerase chain reaction. Immunohistochemical study was performed on paraffin sections. -SMA, FAP, CD68, CD206, CX3CR1, CD45 were used as markers. The area of interlobular veins and interlobular arteries was measured (m2). The number of sinusoidal capillaries and interlobular veins was counted.&#x0D; RESULTS: Based on the results obtained, it is possible to establish the transition point of fibrosis to cirrhosis as an independent separate stage of fibrogenesis. This transition was identified at stage F5, and the process itself from F4/F5 to F6.&#x0D; With the growth of fibrous tissue and nodular restructuring of the liver parenchyma, no progression of dystrophic processes and an increase in the zones of necrosis and necrobiosis of hepatocytes were noted. The number of -SMA+ and FAP+ cells in the period F4F5 did not change (p=0.2073 and p=0.3775, respectively). At the same time, significant F6 cirrhosis was accompanied by an increase in their number by 1.5 times (p 0.00001). Differences in the number of CD68+ cells were revealed only at the F4/F5 stage (2.0 times higher than the control, p 0.00001). The number of CD206+, CX3CR1+ and CD45+ cells remained the same. An increase in the number of interlobular veins (p 0.00001) and a decrease in sinusoidal capillaries (p 0.00001) were found compared to the control.&#x0D; The transition to cirrhosis was characterized by changes in the expression levels of tweak, fn14, ang, vegfa, cxcl12, and mmp-9 mRNAs, as well as the presence and strength of the relationship between them. Significant correlations were revealed between the target genes (r=0.50.84; p 0.01).&#x0D; CONCLUSION: The transition point of fibrosis to cirrhosis has special morphological and molecular genetic features that expand knowledge about pathomorphological changes in the liver.

Comparative morphological characteristics of the uteroplacental area in abnormal placentation

The aim. To carry out a comparative morphological characteristic of the uteroplacental area with abnormal placentation – pl. accreta, pl. increta, pl. percreta. Materials and methods. The study included 47 patients with atypical placentation; the comparison group included 10 healthy pregnant women with uterine scar after a previous caesarean section. A histological study of uteroplacental area samples was performed with hematoxylin and eosin, methylene blue staining. An immunohistochemical study with primary antibodies to cytokeratin 7 (CK7), Hif2a, vascular endothelial growth factor, α-SMA was carried out. The differences between the compared values were considered to be statistically significant at p &lt; 0.05. The results of the study. Pl. accreta was determined in 12 (25.5 %), pl. increta – in 30 (63.9 %), pl. percreta – in 5 (10.6 %) patients. In all patients of the main group, the decidua was completely or partially absent in the area of abnormal placentation or was replaced by an uneven layer of fetal fibrinoid. Cases when placental villi unevenly penetrated into the thickness of myometrium in the form of “tongues” or “coves” bordered by fetal fibrinoid and often located intermuscularly were defined as pl. increta (n = 26). Cases with the placental villi ingrowth to the serous membrane were considered as pl. percreta (n = 5). In cases with deep variants of ingrowth (pl. increta and pl. percreta) (n = 31), the villi were visualized in the lumen of the vessels and the thinning of the lower uterine segment with the presence of stretched muscle bundles was revealed. Aseptic necrosis of the myometrium was found: in 2 (16.7 %) of 12 women with pl. accreta, in 26 (86.7 %) of 30 women with pl. increta and in 5 (100 %) women with pl. percreta. There were no areas of necrosis in the myometrium of the women of comparison group. Conclusion. The appearance and increase of myometrial necrosis zones in response to an increase in the depth of placental villus ingrowth were detected. Myometrial necrosis zones could be the cause of activation of angiogenic factors and an important stimulus for the development of abnormal vascularization in placenta accreta spectrum.

THE PECULIARITIES IN THE FORMATION OF GUNSHOT WOUNDS CHANNELS CAUSED BY 7.62 MM EXPANSIVE «HPBT» AND «SP» TYPE BULLETS

The aim of the study – to study the peculiarities of wound channel formation, causedby 7.62 mm «HPBT» and «SP» bullets, by means of experimental methods of forensicballistics with the use of human biological tissue simulators.Materials and methods. Experimental investigations were carried out under conditionsof the ballistic route of the National Scientific Center «Hon. Prof. M. S. Bokarius ForensicScience Institute». To determine the velocities of the fired bullets, the optoelectronicmeasuring complexes IBX-731.4 were used. Ballistic plasticine «Beschussmasse,6287156» was used as a simulator of human body tissues. The experimental shooting wascarried out with a TS.M308 model .308 «Winchester» sniper rifle. For the experimentalshooting 7.62 × 51 mm (.308 «Winchester») cartridges were used, which were filled with«SP» and «HPBT» bullets.Results. In the course of experimental shooting, the peculiarities of the formation ofdamage channels caused by 7.62 mm «SP» and «HPBT» bullets were determined. Basedon the results of experimental studies, it has been found that the bullets of these types ofthe same caliber and practically the same mass are able to cause different amounts ofbodily injury, which is primarily due to the peculiarities of the design of the bullets. SPtype bullets with a bare core cause more severe bodily injuries, they are less subject tofragmentation and in most cases retain a relatively heavy bottom part. As a consequence,such bullets are able to cause longer wound channels than other types of expansivebullets. HPBT-type bullets can also cause sufficiently large amounts of bodily injury,but because of their design, they are more prone to fragmentation when hitting thehuman body. In the course of the research it has been established that there are severalperipheral (secondary) channels in the area of the main wound channel. They are formeddue to destruction of the bullet and movement of its separate fragments away from themain direction, which leads to complication of diagnostics and further treatment of suchgunshot wounds.Conclusions. The use of 7.62 × 51 mm (.308 «Winchester») expansive «SP» and «HPBT»type bullets in the course of combat operations for firing modern sniper systems resultsto severe bodily injuries. Wounds caused by such bullets are characterized by significantin volume zones of direct traumatic necrosis (contusion), which significantly extend deepinto the tissues and structures of the human body and form even larger in size zonesof conditional necrosis (contusion) and morpho- functional changes. The formationnature of both the main wound channel and the peripheral wound channels, caused byfragments of bullet structural elements, depends on the velocity, design features andposition (projection) of the bullet at the moment of impact. Changes in biological tissuesin the area of wounds caused by these types of expansive bullets have rather complexarchitectonics, which significantly complicates the issue of clear delimitation of gunshotwound zones and adequate surgical intervention. The used investigation methods of thestriking properties of expansive bullets are suitable for predicting the severity and volumeof the caused injuries, as well as for determining the individual characteristic complex ofgunshot wounds signs caused by such bullets.

P02.19.A ASSESSMENT OF HEAT SHOCK PROTEIN HSP70 EXPRESSION IN THE GLIOBLASTOMA STEM-LIKE CELLS

Abstract BACKGROUND Glioblastoma multiforme (GBM) is a malignant brain tumor characterized by poor prognosis. The search of biomarkers for early diagnosis and as targets and for tumor theranostics due to the aggressiveness of the disease is urgently needed. One of the promising targets for therapy of brain tumors is 70 kDa heat shock protein (Hsp70), which has been found on the plasma membrane of various tumor cells (including GBM) but not on normal cells. GBM, like other types of tumors, has cellular heterogeneity. In particular, it contains tumor stem-like cells (TSLCs), which are considered analogues of healthy stem cells and are largely responsible for disease progression, recurrence and mediate resistance to ongoing therapy. In the present work, the histological sections of human GBM were analyzed to identify and quantify the colocalization of the heat shock protein Hsp70 with TSLC markers using the Opal Multiplex analysis system. MATERIAL AND METHODS Histological sections were prepared in accordance with the standard protocol using postoperative material from patients with newly diagnosed GBM (n=15). Zones of necrosis and living tissue were identified on tumor sections. Based on a phenotyping map using the Pathology Views™ fluorescence image analysis software, Hsp70 colocalization was quantified with Nestin and Sox2 in the areas of necrosis and viable tissue employing primary antibodies (anti-Hsp70, anti-Nestin and anti-Sox2). The preparations were stained using an Opal 3-Plex Manual Detection Kit according to the manufacturer's protocol. RESULTS The expression of Hsp70 and TSLC markers (Nestin and Sox2) was detected. The obtained quantitative data did not reveal significant differences in the distribution of cells expressing Hsp70 in the areas of necrosis and viable tissue (36 and 41%, respectively), which may indicate their uniform distribution in the tumor. Sufficiently high colocalization of Hsp70 with Nestin and Sox2 factors was detected mainly in the zone of viable tissue (57 and 30%, respectively). CONCLUSION Colocalization of Hsp70 with TSLC markers indicates the prospects for further studies of Hsp70, as well as its use for targeted therapy of malignant brain tumors.

Comparative study of the hypoglycemic and cardioprotective effects of glucagon-like peptide-1 receptors agonist exenatide, insulin and their combined use in myocardial ischemia/reperfusion injury in rats with experimental type 2 diabetes

Introduction. Insulin therapy used during acute myocardial infarction (AMI) is associated with a high risk of hypoglycemia and high glycemic variability. Therefore, the search for optimal therapeutic approaches which affect both cardioprotection and glycemic correction is relevant. It is known that agonists of glucagon-like peptide-1 (aGLP1) exhibit a protective effect on the myocardium and are involved in the normalization of carbohydrate metabolism. The combined use of аGLP1 exenatide and insulin can be more effective and safe but not insufficiently studied.The aim of this study was the comparative investigation of the effect of exenatide, insulin, and their combination on the volume of myocardial damage and the level of glycemia in the myocardial ischemia/reperfusion (I/RP) model in rats with experimental diabetes mellitus (DM2).Materials and methods. Neonatal streptozotocin diabetes was modeled in male Wistar rats on the 4th–5th day of life. At the age of 3 months, I/RP was fulfilled in animals with confirmed hyperglycemia. Experimental groups were formed depending on the time of therapy with insulin, exenatide, or their combination; before or after ischemia. Ischemia lasted for 40 min, while reperfusion was 120 min. The size of the myocardium necrosis zone and the changes in glycemia level and its variability were determined.Results. Insulin reduced glycemia, but it increased glycemic variability by 60 % and caused hypoglycemia in 32–37 % of animals. Exenatide reduced blood glucose concentration to the level of fasting glycemia and glycemic variability by 1.5–2 times compared with insulin. Exenatide given before ischemia reduced the area of necrosis by 2.2 times. The combined use of insulin and exenatide was accompanied by the absence of hypoglycemia and a decrease in the necrosis zone by 3.2 times compared to the control.Conclusion. The combined use of aGLP1exenatide and insulin in experimental conditions of I/RP+DM2 is the most effective and safe. This makes promising their joint use in patients with DM2, both at high risk of AMI and those undergoing AMI.

PA21 A rare case of diffuse infantile fibromatosis

Abstract Fibromatoses are a heterogeneous collection of tumours of the skin and soft tissue with a large variability in presentation. We present a case of diffuse infantile fibromatosis in a 17-day-old infant boy. He was born at 40 + 7 weeks gestation with a vacuum-assisted delivery following an uncomplicated pregnancy. On physical examination, there were two well-circumscribed circular plaques that coalesced centrally on his left elbow. The plaque measured 2 × 1 cm and was firm, nontethered and nontender. Full skin examination was otherwise normal. Histopathological examination showed hyperkeratotic skin with an attenuated epidermis overlying a dermally based bland-appearing spindle cell proliferation. The lesion was infiltrative throughout the dermis and was arranged in short fascicles and interdigitated with subcutaneous fat, which was extensively trapped by spindle cells. There was no significant atypia and mitotic activity was identifiable but not brisk. Immunohistochemical stains showed strong and diffuse reactivity for smooth muscle actin. Infantile fibromatosis is the childhood counterpart of musculoaponeurotic fibromatosis. It is a benign mesenchymal tumour of infancy and early childhood with diffuse proliferations of oval-to-spindled fibroblasts that infiltrate regional structures. There is a male predominance and in most cases is noted within the first 8 years of life. Clinically, it presents as an asymptomatic, white-yellowish mass that is nontethered to the overlying skin. Although any site may be involved, the majority of cases affect the upper limbs, head and neck region. Histological findings include a wide morphological spectrum. In infants, small oval cells appearing between primitive mesenchymal cells and fibroblasts are in a myxoid background. The cells are loosely arranged and infiltrate skeletal muscle and other regional structures. Peripheral lymphocytic inflammation blends with more cellular proliferation composed of spindle-shaped fibroblasts arranged in bundles and fascicles. There is no well-documented immunohistochemical profile. An important differential diagnosis to exclude is congenital/infantile fibrosarcoma, which, characteristically, has high cellularity, uniform herringbone pattern, high mitotic rate, and zones of haemorrhage and necrosis. In terms of management, as there is a high likelihood of local recurrence in 65% of cases, evidence supports complete excision with documented margins. Where the lesion is diffusely infiltrative, a partial resection to maintain the best postoperative function is recommended.