Vol 161, No 3 (2023)

On March 19, 2024, Evgeniy Ivanovich Chumasov, a well-known neuromorphologist, Doctor of Biological Sciences, and Professor, celebrated his 85^th birthday. Evgeniy Ivanovich is a highly qualified professional in cell biology, cytology, zoology, and histology. Throughout his life, he studied tissue histogenesis and regeneration in the central and peripheral nervous systems. He is a great specialist in nervous tissue cultivation. His works on the development of embryonic anlages of the central and peripheral nervous systems in vitro and after transplantation are still highly relevant and being cited. He authored multiple books and textbooks, several patents, and inventions. Here, we describe the stages of E.I. Chumasov’s scientific life in the field of normal and pathological neurohistology.

BACKGROUND: Dysferlinopathy is heritable progressive muscular dystrophy caused by DYSF mutation. Currently, although skeletal muscle pathology has been defined, only fragmentary and limited myocardium histopathology data are available.

AIM: The study aimed to analyze the pathomorphological status of the myocardium in Bla/J mice models of dysferlinopathy at different ages.

MATERIALS AND METHODS: Data from two experimental groups were analyzed: Bla/J mice with DYSF knockout on 3, 6, and 12 months old and control wild-type Balb/C mice aged 6 months. The expressions and patterns of dyeing of protein dysferlin in the immunofluorescent search method were analyzed. These were held such parameters of the histological characteristic of the myocardium of three dyeing protocols (hematoxylin and eosin, iron hematoxylin by Rego, and hematoxylin-basic fuchsin-picric acid by Lie), and morphometry of the parameters of the cardiomyocytes (length, width of cardiomyocytes, and nuclear perimeter).

RESULTS: The immunofluorescent search method revealed high levels of dysferlin in the myocardium of the control group. Statistical analysis showed significant differences between Bla/J and Balb/C mice: the increasing length and width of cardiomyocytes in dysferlinopathy by 49.9% ((95% confidence interval, 45.9–57.4) and 35.6 (95% confidence interval, 32.9–37.9)), respectively. Nucleus perimeter was significantly reduced in the dysferlinopathy group with disease duration of 6 months (by 23.9 (95% confidence interval, 20.2–27.5) compared with the group with disease duration of 3 months and by 18.8% (95% confidence interval, 8.5–19.7)) and the control group. Consequently, progressive hypertrophy of cardiomyocytes, increasing deformation in cardiomyocytes, intercalated disk destruction, hypoxia features, and necrosis indication were observed, resulting in fibrosis. A pattern of cardiomyocyte size reduction dependent on the aging process was observed.

CONCLUSIONS: Dysferlin deficiency leads to significant damage in the myocardium of Bla/J mice.

BACKGROUND: Radiation hepatitis with the development of radiation-induced acute liver failure is considered one of the most serious complications of radiotherapy for malignant neoplasms of the liver, abdominal organs, or whole body irradiation. However, the exact mechanisms of radiation-induced liver cell death have not been fully elucidated, and therefore the study of changes in the proliferative-apoptotic ratio in liver structures remains relevant, and pre-irradiation administration of ascorbic acid can potentially protect them from the effects of electron irradiation.

AIM: Assessment of proliferation and apoptosis of hepatocytes after administration of ascorbic acid in a model of radiation hepatitis.

MATERIALS AND METHODS: Wistar rats (Rattus Wistar; n=40) were divided into four experimental groups: I — control (n=10); II (n=10) — fractional irradiation with electrons in a total irradiation dose of 30 Gy; III (n=10) — administration of ascorbic acid before electron irradiation; IV (n=10) — administration of ascorbic acid. Animals of all groups were removed from the experiment a week after the last fraction. Morphological and immunohistochemical (with antibodies to Ki-67 and caspase-3) studies were carried out.

RESULTS: A week after electron irradiation, a sharp decrease in the proportion of Ki-67-positive hepatocytes in combination with an increase in immunolabeling with antibodies to caspase-3 was observed in group II. During the administration of ascorbic acid in group III, less pronounced depth and range of liver damage was noted, confirmed by morphological and immunohistochemical methods (less pronounced decrease in the level of Ki-67 expression and an increase in the proportion of caspase-positive hepatocytes compared to the control) methods.

CONCLUSIONS: An immunohistochemical study of proliferation and apoptosis of hepatocytes revealed that a week after fractional electron irradiation in total irradiation dose 30 Gy, there is a decrease in mitotic activity and an increase in cell death, and pre-irradiation administration of ascorbic acid helped level out the detected changes, which indicates its protective effect.

BACKGROUND: Melatonin is a hormone with a wide range of biological activities. The diversity of biological regulatory effects inherent in MT involves this hormone in the formation of adaptive reactions and in the pathogenesis of various diseases. А decrease оf мelatonin secretion due to exposure to light at night is observed in a significant proportion of people. A number of previous studies have shown that melatonin deficiency, causes significant changes in the structure of the liver of laboratory animals. The state of ultrastructural features of hepatocytes, and in particular their mitochondria, under conditions of dark deprivation remains poorly understood.

AIM: To study the ultrastructural features of liver hepatocytes of male Wistar rats under conditions of 21-day dark deprivation.

MATERIALS AND METHODS: The study was performed on 40 male Wistar rats, divided into 2 groups: group 1 was kept under a fixed light regime; group 2 was kept under dark deprivation conditions for 24 h a day. Liver samples, were analyzed using a transmission electron microscope. Micromorphometric methods were used to assess the mitochondrial apparatus of hepatocytes. Statistical processing of the results was performed in the GraphPad Prism v. 8.4.1 program (GraphPad, USA).

RESULTS: In hepatocytes of rats of II group, dark deprivation causes a transformation in the shape of the nuclei, accompanied by swelling of the cytoplasm and the presence of a significant number of lipid-containing vacuoles. Mitochondria are characterized by pronounced hyperplasia, size polymorphism, high electron density, and disordered cristae orientation. In the cytoplasm, the phenomenon of shedding of ribosomes from the endoplasmic reticulum is observed. The number of glycogen granules is significantly reduced. The studied micromorphometric parameters of mitochondria are significantly reduced relative to the control.

CONCLUSIONS: The study suggests that melatonin deficiency, resulting from dark deprivation, leads to a number of significant ultrastructural changes in hepatocytes, especially their mitochondrial apparatus.

BACKGROUND: It is known that intercellular communications in the peripheral nervous system are provided by various types of intercellular contacts, in particular, gap junctions formed by connexin proteins. The literature contains data on changes in the expression of connexin-32, connexin-46 and other types of connexins during nerve injury. However, very few studies describe changes in connexin-43 expression in similar pathologies.

AIM: The purpose of this study was to study the distribution and localization of the gap junction protein connexin-43 (Cx43) in cells of intact and injured rat sciatic nerve.

MATERIALS AND METHODS: Damage to the sciatic nerves of Wistar rats of the experimental group (n=5) was carried out by nerve ligation for 40 s. Animals without damage of sciatic nerve were studied as a control group (n=5). Immunohistochemical detection of Cx43 was performed on paraffin sections.

RESULTS: It has been established that the protein is contained in the cells of the perineurium and epineurium of both the intact nerve and after the application of a ligature. In the area of the endoneurium, in the absence of nerve damage, Cx43 is detected only in the endotheliocytes of a few vessels. In the endoneurium of the injured nerve, a large number of large Cx43-immunopositive cells with processes were identified.

CONCLUSIONS: It has been established that Cx43-containing cells are identified in the endoneurium of the sciatic nerve only after damage. To clarify whether such cells belong to a specific cell type, additional studies are necessary.

BACKGROUND: Somatotyping has a long history and at the same time is widespread today in medicine, sports, and biomedical research, which necessitates the analysis of accumulated scientific facts.

AIM: Identify the main trends and patterns of development of methodological approaches to determining somatotypes based on the analysis of domestic scientific publications from 2013 to 2023.

MATERIALS AND METHODS: Based on scientific publications indexed in the RSCI, a database has been formed on the use of somatotype assessment methods for the period from 2013 to 2023. Keywords of publications, frequency and scientific areas of application of somatotyping schemes were analyzed.

RESULTS: Anthropometry techniques were most often used to identify predictors of adaptation failure or disease development. When searching for characteristics of samples that differ in influencing factors, body composition was more often used. An increase in studies using somatotyping according to B.H. Heath and J.E. Carter (hereinafter — Heath–Carter) and M.V. Chernorutsky has been revealed. The share of studies performed using the methods of R.N. Dorokhov and V.G. Petrukhin and J.M. Tanner is decreasing, and the variety of somatotyping schemes used. In the study of adolescence, first adulthood and when somatotyping women, somatotyping according to R.N. Dorokhov and V.G. Petrukhin was more often used, in clinical studies — according to W.L. Rees и H.J. Eysenck, in the study of athletes — according to Heath–Carter, in combined somatotyping — according to P.N. Bashkirov, J.M. Tanner and W.L. Rees and H.J. Eysenck.

CONCLUSIONS: The main problem, that anthropologists faced was the comparability of the results obtained using different somatotyping schemes.

BACKGROUND: The critical factor when working with immunohistochemistry in laboratory animals is to select appropriate secondary antibodies, that allow clear and specific visualization of tissue antigens. Many reliable secondary reagents are currently not available for purchase, which determines the high relevance of replacing them with other detection systems.

AIM: To verify the effectiveness of available secondary reagents for immunohistochemical research of the rat brain.

MATERIALS AND METHODS: Brain samples from Wistar (n=2) and SHR (n=2) were used for the study. Iba-1, GFAP and vimentin immunohistochemistry was carried out using various polymer-based detection systems, namely UltraVision Quanto Detection System HR, N-Histofine Simple Stain MAX PO and UnoVue Rabbit HRP.

RESULTS: All three studied polymer systems demonstrated visualisation of target proteins in brain tissues and cells corresponding to the general understanding of structures containing Iba-1, GFAP and vimentin. The UnoVue Rabbit HRP and UltraVision Quanto Detection System HRP kits showed good and similarly specific immunohistochemical reaction. However, the UnoVue Rabbit HRP kit was less sensitive compared to the UltraVision Quanto Detection System HRP. The reaction with N-Histofine Simple Stain MAX PO was not optimal due to the presence of non-specific background staining that was not present with other reagents. Apparently, this is due to the focus of the kit on immunohistochemical staining of human tissue and the likely absence of an antibody purification with rat serum.

CONCLUSIONS: Two secondary antibody kits from the three studied showed optimal efficiency of immunohistochemical reaction and minimal background staining. N-Histofine Simple Stain MAX PO is not suitable for immunohistochemical research of rat brain tissue.

Tumor-infiltrating lymphocytes are currently considered as a prognostic biomarker of cancer patient survival and response to therapy, as well as a target for immunotherapy. However, tumor-infiltrating lymphocytes are only part of the tumor microenvironment, which consists of cellular and cytokine components. The cellular component also includes tumor-associated macrophages, neutrophils, fibroblasts and other cells. The cytokine component is represented by the products of the activity of all cells of the microenvironment, as well as tumor cells.

This review examines various subpopulations of tumor-infiltrating lymphocytes, shows their interaction with tumor cells and other cell populations of the tumor microenvironment, and also describes the potential for clinical use of this cell population as a biomarker for predicting clinical outcomes and the possibility of their use to determine the most effective treatment for breast cancer.

Obtaining new data on the previously poorly studied components of the tumor microenvironment and tumor-infiltrating lymphocytes, as well as the formation of ideas about the mutual influence of tumor microenvironment cells and tumor cells open up the prospect of developing innovative models of therapy. One of the modern directions is the search for molecular mechanisms of maintaining the antitumor activity of tumor-infiltrating lymphocytes, as well as the polarization of cells in the tumor microenvironment. Advances in the field of molecular oncology will make it possible to reduce the mortality rate of cancer patients to minimal levels while maintaining or even improving their standard of living.

In the article authors analyze the participation of domestic and foreign morphological scientists in the activities of the Imperial St. Petersburg Academy of Sciences. From the founding of the Academy (1724) until 1917, a number of outstanding morphologists were elected to its full members, including K.F. Wolf, A.E. Protasov, Kh.G. Pander, K.E. von Baer, A.O. Kovalevsky, F.V. Ovsyannikov, A.S. Dogel, D.N. Anuchin. The scientific works of these scientists constituted the glory of Russian science. The academy’s connections with foreign scientists also developed, many of whom were elected corresponding members and foreign honorary members of the academy, including a number of morphologists, comparative anatomists and histologists. Thus, the following were elected as foreign honorary members: in 1737 — R.A. Reaumur, in 1776 — A. von Haller, in 1802 — J. Cuvier; in 1826 — J.W. von Goethe; in 1905 — C. Golgi. Correspondents were elected as foreign members: in 1826 — R. Brown, in 1827 he became a full member of the academy; in 1826 — K.E. von Baer, who after moving to Russia became its ordinary academician (1827), and then, after his resignation, its honorary member (1862); in 1832 — I.P. Muller; in 1836 — Ya.E. Purkinje; in 1850 — M.Ya. Schleiden; in 1857 — R.A. Kolliker; in 1881 — R. Virchow; in 1897 — F. von Leydig.

Communication between foreign scientists and the Academy was carried out in several directions, the main of which was sending their works to the Academy and publishing their works in Academy publications. Another direction was the training of Russian students, as well as internships for Russian researchers preparing for a professorship.