[Exosomes' role in intercellular interactions in different variants of lung injury in fatal cases of COVID-19]. / Rol' ekzosom v mezhkletochnykh vzaimodeistviyakh pri razlichnykh variantakh porazheniya legkikh v letal'nykh sluchayakh COVID-19.

BACKGROUND: Extracellular vesicles are surrounded by a phospholipid bilayer, carrying various active biomolecules and participating in many physiological and pathological processes, including infectious ones. OBJECTIVE: To research the role of exosomes in intercellular interactions in the pathogenesis of various types of lung damage in fatal cases of COVID-19. MATERIAL AND METHODS: We conducted a clinical and morphological analysis of 118 fatal cases caused by coronavirus infection in Moscow. We selected 32 cases with morphological signs of various types of lung lesions for immunohistochemical reaction (IHC) with antibodies against tetraspanin proteins (CD63, CD81), which are involved in the assembly of exosomes, as well as with antibodies against viral proteins: nucleocapsid and spike protein. We determined the main producing cells of extracellular vesicles and cells containing viral proteins, carried out their comparison and quantitative analysis. RESULTS: IHC reaction with antibodies against CD63 showed cytoplasmic granular uniform and subapical staining of cells, as well as granular extracellular staining. We determined similar staining using antibodies against viral proteins. Extracellular vesicles were found in the same cells as viral proteins. The main producing cells of vesicles and cells containing viral proteins were found to be macrophages, type II pneumocytes, and endothelial cells. CONCLUSION: Taking into account the results of the literature, the localization of viral proteins and extracellular vesicles in the same cells indicates the key role of vesicles in the pathogenesis of various forms of lung damage by the SARS-CoV-2 virus, in the dissemination of the pathogen in the organism, which leads to interaction with the adaptive immune system and the formation of immunity.

[Morphogenetic and pathogenetic features of hypertrophic and keloid scars of the head and neck]. / Morfogeneticheskie i patogeneticheskie osobennosti gipertroficheskikh i keloidnykh rubtsov golovy i shei.

OBJECTIVE: Evaluate the morphogenetic and pathogenetic features of hypertrophic and keloid scars of the head and neck. MATERIAL AND METHODS: The study included 286 patients, among them 176 (61.5%) patients with hypertrophic and 110 (38.5%) with keloid scars aged 18 to 65 years with a disease duration from 1 month to 2 years. Material for histological and immunohistochemical (IHC) studies of scar tissue was fixed in 10% buffered formalin. Serial paraffin sections were stained with H&E, according to Van Gieson and Weigert. IHC was performed using monoclonal mouse antibodies to collagen type I (clone 3G3, Santa Cruz, dilution 1:100), collagen type III (clone B-4, Santa Cruz, dilution 1:50), collagen type IV (clone COL-94, Santa Cruz, dilution 1:50), MMP-1 (clone 3B6, Santa Cruz, dilution 1:100), α-SMA1 (clone 1A4, Dako Agilent, dilution 1:100) and rabbit polyclonal anti-TGFß antibodies (clone 3C11, Santa Cruz, 1:100 dilution). RESULTS: Pathogenetic, morphological and immunohistochemical differences in hypertrophic and keloid scars were established depending on their degree of maturity. In the formation of hypertrophic scars, the key factor in sclerotic processes is TGF-b on the background of low MMP1 activity. Keloid scars were distinguished not only by the accumulation of hard-to-degrade collagens, but also by the development of an osteoclast-like reaction with a high content of MMP1. Immature scar tissue was characterized by the presence of myofibroblastic α-SMA1 positive focus and center of inflammatory changes. CONCLUSIONS: The data obtained allow substantiating new approaches to the treatment of patients with hypertrophic and keloid scars.

[The role of expression of monocarboxylates of the first and fourth types (MCT1, MCT4) by tumor and stromal cells of prostate cancer in determining the prognosis and the efficiency of definitive treatment].

AIM: A search for new methods for diagnosing clinically significant prostate cancer is of importance due to the insufficient accuracy of modern methods in detecting aggressive tumors. One of the promising opportunities for the early diagnosis of clinically significant prostate cancer is the assessment of the glycolytic profile of the tumor by determining the expression of monocarboxylates (MCT) types 1 and 4 in tumor cells, as well as in adjacent stromal cells. MATERIALS AND METHODS: An analysis of patients of who underwent radical prostatectomy at the Institute of Urology and Reproductive Health of Sechenov University from 2015 to 2017 was carried out. The patients with histologically confirmed prostate adenocarcinoma were included in the study. Among them, the presence or absence of biochemical recurrence during the first year was studied. An immunohistochemical (IHC) study of postoperative specimen was performed to determine the expression of MCT1 and MCT4 by tumor and stromal cells. The correlation between the intensity of their expression and the risk of biochemical recurrence and the tumor characteristics was evaluated. RESULTS: High membrane expression of MCT1 directly correlated with high stromal expression of MCT4 (r=0.314, p<0.003). A significant direct correlation was found between the predominance of stromal expression of MCT4 over membrane expression and biochemical recurrence (r=0.403, p<0.001), as well as a high ISUP group (4 and 5) (r=0.294, p=0.005). CONCLUSIONS: Determination of the level of expression of type 1 and 4 monocarboxylate transporters in adenocarcinoma cells and tumor stromal cells can become an effective tool for risk stratification, and may also predict the biological behaviors of the prostate cancer and the efficiency of definitive treatment.

[Clinical and morphological characteristics of SARS-CoV-2-related myocarditis proven by the presence of viral RNA and proteins in myocardial tissue]. / Kliniko-morfologicheskaya kharakteristika SARS-CoV-2-assotsiirovannogo miokardita, podtverzhdennogo nalichiem RNK i belkov virusa v tkani miokarda.

OBJECTIVE: To investigate the clinical and morphological features of SARS-CoV-2-related myocarditis, by determining the presence of viral RNA and proteins in myocardial tissue. MATERIAL AND METHODS: The study was conducted to examine the material of 32 autopsies with a confirmed diagnosis of myocarditis. There were data of a morphological study, including a standard histological study, as well as immunohistochemical determination of the surface markers CD45, CD3, CD20, and CD68 cells of an inflammatory infiltrate and virus proteins (SARS-CoV-2 nucleocapsid protein and spike protein). Positive and negative control tests were carried out. In addition, coronavirus RNA was detected in the myocardium using a polymerase chain reaction. RESULTS: Polymerase chain reaction (PCR) revealed viral RNA in myocardial tissue. Viral proteins were identified in the macrophages of an inflammatory infiltrate and cardiomyocytes. CONCLUSION: The findings may suggest that the virus persists in the myocardium and chronic myocarditis develops.

[Lymphocytic myocarditis in patients with COVID-19 (4 autopsy cases)]. / Limfotsitarnyi miokardit u patsientov s COVID-19 (4 autopsiinykh nablyudeniya).

The paper describes 4 autopsy cases of myocarditis in elderly patients with confirmed coronavirus infection. It gives the data of a morphological study of heart specimens and a detailed characterization of a myocardial infiltrate. An immunohistochemical study with cellular infiltrate typing was performed. The fact that lymphocytic viral myocarditis can develop in COVID-19 was morphologically and immunohistochemically confirmed. The features of myocarditis in COVID-19 are the development of the former in the presence of coronaritis and the possibility of its concurrence with lymphocytic endo- and pericarditis.

Morphological and Molecular-Biological Features of Inflammatory and Regeneratory Processes in Peridont Tissues with Periimplantitis and Periodontitis.

A morphological and immunohistochemical study of periodontal tissues was performed in patients with chronic generalized periodontitis who underwent dental implantation. We studied 44 biopsy specimens from 21 patients (13 women and 8 men) aged 35-60 years with a diagnosis of periimplantation mucositis (7 patients), periimplantitis (8 patients), and severe chronic generalized periodontitis (6 patients). It was established that periimplantitis differs from periimplantation mucositis by a more pronounced inflammatory reaction with a clear predominance of plasma cells in the infiltrate, which captures the fibrous capsule around the implant, destroys it and further spreads to the bone tissue. Based on the immunohistochemical differences in SMA, VEGF, and Ki-67, it is concluded that periimplantation mucositis and periimplantitis are successive stages of progression of the same process. When comparing chronic generalized periodontitis and periimplantitis, the latter shows much more pronounced inflammatory and destructive processes in the area of the implant, due to the addition of immune inflammation, impaired regeneration processes, and destruction of bone tissue.

[Pathomorphological features of the placentas and the placental sites after in vitro fertilization with a donor egg]. / Patomorfologicheskie osobennosti platsent i platsentarnykh ploshchadok posle ékstrakorporal'nogo oplodotvoreniia s primeneniem donorskoi iaitsekletki.

OBJECTIVE: To identify the structural and immunohistochemical features of the placentas and the placental sites after in vitro fertilization (IVF) with a donor egg (surrogate motherhood). SUBJECT AND METHODS: Morphological and immunohistochemical studies were performed on the placental (a placental disk) and placental bed materials obtained after caesarean delivery. The investigation enrolled 26 patients whose pregnancy occurred with IVF with a donor egg according to the surrogacy (IVF-S) program. A comparison group included 13 patients whose pregnancy occurred after IVF with their own eggs. An immunohistochemical study was conducted on paraffin sections made from biopsy material; mouse antibodies to total cytokeratin (clone AE1/AE3, 'Dako'), HLA-DR (clone TAL.1B5, 'Dako'), and CD138 (clone MI15, 'Dako') were used as primary antibodies. RESULTS: The histological examination of the placentas in the IVF-S group showed the high incidence of central ischemic heart attacks (69%), dissociated cotyledon development (61%), pathological villous immaturity mainly with the predominance of intermediate differentiated villi (46%), and massive perivillous fibrinoid deposition (73%). The obtained differences between with the study and comparison groups were significant (p<0.05). The IVF-S group was characterized by the development of lymphoplasmacytic deciduitis (1.23±0.4 and 0.5±0.3 scores). Examination of the placental site biopsy material in the IVF-S group revealed the following changes: remodeling of the spiral arteries was incomplete in more than 40% of cases, and in 30% of the spiral arteries had no gestational changes. In the comparison group, more than 90% of the spiral arteries were characterized by complete remodeling during pregnancy. There was also an increase in the count of multinucleated trophoblastic giant cells (104.56±4.21 and 65.67±14.45) and HLA-DR positive cells (41.86±5.32 and 29.00±1.87). CONCLUSION: The placentas and the placental sites of the women whose pregnancy occurred with IVF-S are characterized by the development of high lyoplasmacytic deciduitis activity and pronounced placental immune alterations manifested by the high incidence of immune responses at the sites of the closest contact between maternal and fetal tissues. The placental bed exhibited defective spiral artery remodeling, development of chronic inflammatory lesions in the perivascular areas, and an increase in the counts of HLA-DR positive cells and multinucleated trophoblastic giant cells.

Correlation of synovial caspase-3 concentration and the photodynamic effectiveness in osteoarthritis treatment.

BACKGROUND: The present study focuses on investigation of Intra-articular PDT mechanisms for OA treatment. Also, a search for determination of the most effective dose of chlorin e6 (Ce6) for anti-inflammatory PDT of OA was carried out. METHODS: The study was carried out on laboratory animals (11 Chinchilla rabbits, 1 year, 2.5 kg) with a gonarthritis model of post-traumatic OA. According to the instructions for using Photoditazin (Ce6 based PS) for PDT of human oncological and non-oncological diseases, the recommended dose is 0.7-1.2 mg/kg. For studies on rabbits, taking into account the conversion coefficient (3.2), the PS doses of 2.4, 3.2 and 6.4 mg/kg were selected. Fluorescence spectra were measured intra-articular before and after PDT using spectrometer with fiber-optic probe. The intrajoint PDT was carried out using a laser (662 ± 10 nm) and a fiber-optic catheter with a cylindrical diffuser inside a sapphire needle for a uniform distribution of the laser radiation. The immunohistochemical study was carried out by staining the samples with caspase-3. RESULTS: Histological and immunohistochemical analysis showed that the best PS dose for intravenous administration for PDT of rabbit gonarthritis is 3.2 mg/kg. The PS concentration directly in the synovial tissue was 0.5 mg/kg, and this was enough to achieve the most positive results to reduce the caspase-3 level. CONCLUSION: The caspase-3 level correlates well with other signs of inflammation in the synovial membrane (edema, etc.). Therefore, to assess the PDT effectiveness in the treatment of gonarthritis accompanied by synovitis, it is sufficient to analyze only for caspase-3. The efficacy of PDT with Ce6 showed that 3.2 mg/kg PS dose (1 mg/kg for a human) is the most effective.

Immunohistochemical and Morphological Study of Periodontal Tissues in Predicting the Results of Dental Implantation in Patients with Chronic Periodontitis.

Morphological and immunohistochemical examination was made on 24 gum biopsies obtained from 35- to 60-year-old patients with diagnosis of partial secondary adentia, chronic generalized moderate to severe periodontitis (19 patients), as well as on the biopsy samples from five patients without pathological periodontal changes who underwent dental implantation. Serial paraffin sections were treated with antibodies against Ki-67, VEGF, and SMA. In patients with severe chronic periodontitis, a high proliferative activity of epithelium indicative of hyperplastic changes was observed, as well as a reduced number of the SMA-positive cells and actual absence of the SMA-positive cell couplings associated with the "growth zones" in tissues, which testifies indirectly to a lower tissue regenerative capacity. Hence, before dental implantation, additional anti-inflammatory and pro-regenerative treatment is required.

[Growth mechanisms and morphological structural features of large uterine leiomyoma]. / Mekhanizmy rosta i osobennosti morfologicheskogo stroeniia leiomiomy matki bol'shogo razmera.

OBJECTIVE: To investigate the growth mechanisms of large uterine leiomyoma (LULM) on the basis of a clinical morphology examination, by providing immunohistochemical (IHC) characteristics of the expression of growth factors (transforming growth factor-beta (TGFß) and platelet-derived endothelial cell growth factor (PD-ECGF)) and markers of stemness (CD117/c-kit, Connexin 43, Nestin) and proliferation (Ki-67). SUBJECT AND METHODS: The investigators examined surgical specimens from 38 women diagnosed with simple uterine leiomyoma (ULM), who had been divided into two groups: 1) 21 patients with LULM (>6 cm in diameter) (a study group); 2) 17 patients with small ULM (<4 cm in diameter) (a comparison group). Each group was also divided into two age subgroups (younger (<45 years) and older (≥45 years) subgroups (1a (n=12), 1b (n=9), 2a (n=8) and 2b (n=9), respectively. Histological specimens were used to make IHC examination with antibodies against TGFß, PD-ECGF, CD117/c-kit, Connexin 43, Nestin, and Ki-67. RESULTS: The growth mechanisms of LULM of simple histological structure were found to be associated with the larger number of growth zones in the tumors, with their enhanced cellular proliferative activity, and with the appearance of cells with signs of stemness, which is combined with the preserved subsequent maturation of tumor cells and determines the benign nature of LULM. CONCLUSION: There were differences in the molecular profile of LULM and small ULM, as well as LULM in perimenopausal and young women by the expression levels of Ki-67, TGFß, PD-ECGF, CD117, and Connexin 43, which can be used for diagnosis, prediction, and development of targeted therapies.

[Small-cell rectal carcinoma coexisted with tubular-villous adenoma].

The paper describes a case of small-cell high-grade endocrine carcinoma of the rectum concurrent with tubular-villous adenoma in a 58 year-old woman. All epithelial cells of adenoma have been ascertained to be "mixed" exoendocrine endocrinocytes (secrete mucus and contain neuroendocrine granules), contain progesterone receptors, and express CK 8, 18, and 19. Some high-grade neuroendocrine carcinoma cells also secrete mucus. The presence of mitotic activity (Ki-67, 35%), high p53 protein expression (more than 50%), marked CK 18 expression, and clinical data confirm high malignancy of this histological type of the tumor.

[Macrosystemic approach in cytological studies of the urothelial urinary bladder using the computerized morphometry data].

The work is devoted to studying a possibility to use, for cytological diagnostics of urogenital neoplasms, of macrosystemic approaches, aimed to estimate cell systemic reactions during blasttransformation and treatment (exemplified by 95 urinary bladder cancer patients). Changes in size of nuclear and cytoplasmic areas were studied, in addition to the average activity of synthetic processes or functional activity of cell structures, and to the level of heterogeneity of functional tension in the cell space.