[Analysis of the work of the Immunohistochemical Quality Control Center of the Russian Medical Academy of Continuous Professional Education of the Ministry of Health of Russia in 2023]. / Analiz raboty Tsentra kontrolya kachestva immunogistokhimicheskikh issledovanii FGBOU DPO «RMANPO¼ Minzdrava Rossii v 2023 godu.

The article demonstrates a detailed analysis of the results of the rounds of quality control of immunohistochemical studies conducted by the Central Committee of the Immunohistochemical Quality Control Center of the Russian Medical Academy of Continuous Professional Education of the Ministry of Health of Russia in 2023. Typical shortcomings and errors in the immunohistochemical examination of various tumors have been identified and ways to eliminate them are given. Particular attention is paid to defining a panel of standard breast cancer markers and eliminating the shortcomings of immunohistochemical examination of markers of accompanying diagnosis.

[Comparative immunohistochemistry study of different antibodies clones for detection of breast cancer markers (estrogen receptor, progesterone receptor, HER2/neu, Ki-67)]. / Sravnitel'noe issledovanie antitel razlichnykh klonov dlya vyyavleniya markerov raka molochnoi zhelezy (retseptorov estrogenov, progesterona, HER2/neu, Ki-67) immunogistokhimicheskim metodom.

OBJECTIVE: A comparative study of detection of breast cancer markers (estrogen receptors, progesterone receptors, HER2/neu, Ki-67) by immunohistochemical method with antibodies produced by PrimeBioMed (Russia) and antibodies produced by Roche Ventana (USA). MATERIAL AND METHODS: Surgical specimens and biopsies from 37 patients with invasive breast cancer were used. Sections were stained with antibodies of clones ER SP1 and GM030, PR 1E2 and PBM-5B8, HER2/neu 4B5 and PBM-46A6, Ki-67 30-9 and GM010. RESULTS: There was a high positive and significant correlation between the immunohistochemistry results and antibodies of the clones ER-SP1 and GM030, PR1E2 and PBM-5B8, HER2/neu4B5 and PBM-46A6, Ki-67 30-9 and GM010. CONCLUSION: The study showed the possibility of using antibodies of clones GM030, HER2/neu 4B5, PBM-46A6, GM010 (PrimeBioMed) on the Ventana Bench Marck Ultra automatic immunostainer using the detection system UltraView Universal DAB Detection Kit.

[Assessment of HER2 status of carcinomas of various localizations]. / Opredelenie HER2-statusa kartsinom razlichnykh lokalizatsii.

A detailed description of the methodological aspects of the evaluation of HER2-status in carcinomas of such localizations as the mammary gland, pancreas, salivary glands, stomach, colon, endometrium, bladder, lungs is presented. Approaches and criteria for assessing HER2 status from methodological and clinical points of view are analyzed. The data are systematized in tables for use in practical diagnostic work.

[Predictive markers of immunotherapy in cervical cancer]. / Prediktivnye markery immunoterapii v rake sheiki matki.

OBJECTIVE: Study of PD-L1 expression in squamous and adenosquamous cell cervical cancer (CC) by immunohistochemical (IHC) method, assessment of the relationship between PD-L1 tumor status and its clinical and morphological characteristics, TILs, MSI/dMMR, and HPV tumor status. MATERIAL AND METHODS: Surgical material was obtained from 41 patients with CC, on which the expression of PD-L1, proteins of the MMR system and p16 was studied by the IHC method, the TILs index was determined. RESULTS: Positive PD-L1 status was found in 51.2% of the studied CC samples. In the study sample, the level of PD-L1 expression depended on the severity of lymphoid infiltration of the tumor (p=0.038), it was shown that a positive PD-L1 status of CC can be expected with a TILs value greater than or equal to 50%. The age of the patients, the histological variant of the tumor, the pT and pN stage, the presence of lymphovascular invasion, and the HPV status did not statistically significantly affect the level of PD-L1 expression, however, there was an association between the PD-L1 status and the grade of CC malignancy (p=0.027). The presence of the MSI/dMMR phenomenon was detected in a small percentage of carcinomas (4.9%), the PD-L1 status of these tumors was determined as positive. CONCLUSION: A positive PD-L1 status is determined in a significant number of cases of CC, regardless of most of the studied clinical and morphological characteristics; there is a statistically significant relationship between PD-L1 expression and the degree of tumor differentiation and TILs. It has been shown that CC with the MSI/dMMR phenomenon is characterized by a positive PD-L1 status. The authors consider it necessary to study the expression of PD-L1 in patients with cervical carcinomas in order to determine the possibility of prescribing personalized therapy with immune checkpoint inhibitors.

[Updated CAP guidelines for determining molecular biological subtypes of breast cancer]. / Obnovlennye rekomendatsii CAP po opredeleniyu molekulyarno-biologicheskikh podtipov raka molochnoi zhelezy.

Updated 2023 guidelines from the College of American Pathologists (CAP) on immunohistochemical detection of human epidermal growth factor receptor type 2 (HER2), receptors of estrogen (ER) and progesterone (PgR), and the cell proliferation marker Ki-67 in breast cancer are presented. Attention is drawn to the emergence of two new terms «ER Low Positive¼ and «HER2 Low¼ to characterize tumors with low expression of estrogen receptors and HER2.

[Investigation of the mutational status of the FGFR3 gene in urothelial bladder carcinoma]. / Issledovanie mutatsionnogo statusa gena FGFR3 v urotelial'noi kartsinome mochevogo puzyrya.

OBJECTIVE: To study the somatic mutational status of the FGFR3 gene in urothelial bladder cancer (BC) and evaluate its relationship with the clinical and morphological characteristics of the tumor, deficiency of the DNA mismatch repair (dMMR), PD-L1 tumor status, and immunohistochemical (IHC) expression of the p16 protein. MATERIAL AND METHODS: Surgical material of 40 patients with BC, on which the mutational status of the FGFR3 gene was studied using the molecular genetic method, as well as the MMR status, PD-L1 and p16 expression by the IHC method. RESULTS: FGFR3 mutations, such as G370C, S249C, S371C/Y373C, R248C, were detected in 35.0% of the studied BC samples. FGFR3 status did not depend on the gender and age of patients, as well as on the degree of tumor lymphoid infiltration (TILs). Statistically significant differences were found in the analysis of FGFR3 status depending on the histological structure and degree of tumor differentiation, as well as on the pT stage. The FGFR3 status of BC was not associated with the IHC expression of the studied proteins of the MMR system, as well as with the PD-L1 status. Higher levels of PD-L1 expression were demonstrated by BC tumor cells, in which no aberrations in FGFR3 were detected. There was no significant association between p16 status and the presence of FGFR3 mutations, but for FGFR3-positive carcinomas, the basal pattern of p16 staining by IHC was noted. CONCLUSION: A positive somatic mutational status of the FGFR3 gene was statistically significantly more common in the group of papillary low-grade non-muscle-invasive BC, demonstrating basal p16 IHC staining. In the study sample, there was no statistically significant relationship between the FGFR3 status of BC and gender and age differences, TILs, MMR status, PD-L1 status (SP142 and 22C3), and p16 status. The results of the study indicate the need to determine the FGFR3 status in patients with BC for further prescription of personalized therapy.

[External quality control of immunohistochemical studies in pathomorphology: tasks, problems, solutions, development prospects]. / Vneshnii kontrol' kachestva immunogistokhimicheskikh issledovanii v patomorfologii: zadachi, problemy, puti resheniya, perspektivy razvitiya.

In 2022, the Quality Control Center for Immunohistochemical Studies of the Russian Medical Academy of Continuing Professional Education conducted 12 rounds of markers for breast, lung, prostate, bladder cancer with the participation of 83 laboratories. For the first time, a round was held to control the method of in situ hybridization in the diagnosis of breast cancer, and a digital round. Typical problems in carrying out immunohistochemical studies in oncomorphology have been identified and the importance of participation of laboratories in external control has been shown.

[Results of the work of the Russian Medical Academy of Continuous Professional Education Center for quality control of immunohistochemical studies for 2019-2021]. / Itogi raboty Tsentra kontrolya kachestva immunogistokhimicheskikh issledovanii FGBOU DPO «RMANPO¼ za 2019­2021 gg.

In 2019-2021 the Russian Medical Academy of Continuous Professional Education Center for quality control of immunohistochemical studies conducted rounds on the most used tumour markers of various localizations. The deficiencies in the conduct of immunohistochemical studies were identified and the importance of the participation of medical organizations in measures to improve the quality control of immunohistochemical studies in oncomorphology was shown.

[Case of peripartum cardiomyopathy misdiagnosed as pulmonary changes due to COVID-19].

A 37-year-old female patient was admitted 16 days after delivery in a hospital for infectious diseases with cough, shortness of breath, and infiltrative changes in the lungs that were interpreted as viral pneumonia. Considering the failure of therapy and the history, peripartum cardiomyopathy was suspected. Examination revealed a decrease in left ventricular ejection fraction to 30 %, ultrasonic signs of lung congestion and bilateral hydrothorax. The patient was diagnosed with peripartum cardiomyopathy accompanied by functional class 4 heart failure. A specific feature of this case was fast positive dynamics with complete regression of the clinical picture of congestion and improvement of the left ventricular myocardial function associated with the treatment.

[Reed nevus and cutaneous melanoma in children: an immunohistochemical study]. / Nevus Rida i melanoma kozhi u detei. Immunogistokhimicheskoe issledovanie.

Certain difficulties arise in the differential diagnosis between Reed nevus that is common in children and adolescents and cutaneous melanoma that is extremely rare in patients of this age group. In addition to the classical histological examination, an immunohistochemical test for marker proteins is carried out to improve the accuracy of the diagnosis of pigmented skin neoplasms. OBJECTIVE: To evaluate the suitability of a set of proteins (cyclin D1, p16INK4a, and HLA class I antigens) in the differential diagnosis of Reed nevus and cutaneous melanoma in children and adolescents. MATERIAL AND METHODS: Three samples of pigmented skin neoplasms were taken during surgical treatment in patients at the Blokhin National Medical Research Center of Oncology: two samples of Reed nevus (from a 9-year-old girl and a 15-year-old boy) and a sample of cutaneous melanoma that had developed in an 8-year-old boy. The materials were presented by paraffin blocks. Immunohistochemical staining was performed using antibodies against cyclin D1, p16INK4a, and HLA class I antigens. RESULTS: Differences were found between nevus and melanoma in the expression of all three markers. Thus, the nuclear expression of cyclin D1, a proliferation activator, occurred in some cells in the nevus samples and in the vast majority of cells in the melanoma samples. The nuclear expression of the cell cycle inhibitor p16INK4a was dramatically reduced in melanoma compared to nevus. HLA class I antigens were detected on the surface of individual nevus cells, but were completely absent on the membranes and in the cytoplasm of melanoma cells, which could promote the evasion of this tumor from the body's immunological supervision. CONCLUSION: Immunohistochemical test for the proteins cyclin D1, p16INK4a, and HLA class 1 antigens is a promising approach to differentially diagnosing between Reed nevus and cutaneous melanoma in children and adolescents.

[Immunohistochemical study of MSI markers in breast cancer]. / Immunogistokhimicheskoe issledovanie markerov MSI pri rake molochnoi zhelezy.

OBJECTIVE: To investigate the expression of microsatellite instability (MSI) markers, which is detected by an immunohistochemical technique, and to compare the expression with the PD-L1 status in luminal B, HER2-negative and triple-negative breast cancer. MATERIAL AND METHODS: The investigation included tumors from 40 patients with triple-negative and luminal B, HER2-negative subtypes. Immunohistochemical study was performed using Ventana antibodies: anti-MLH1 (clone M1), anti-MSH2 (clone G219-1129), anti-PMS2 (clone A16-4), and anti-MSH6 (clone SP93). MSI was assessed according to the standard criteria. RESULTS: The PD-L1-positive status was present in 14 (35%) of the 40 patients. Moreover, MSI-H was detected in only 1 (2.50%) case. The two-year survival rate was 87.5%; it should be noted that the median survival rate was not reached either in the study sample or in the groups divided according to PD-L1 and MSI statuses. The overall survival rate for patients with MSI was 75% (3/4). CONCLUSION: The first comparative study of the expression of PD-L1 and immunohistochemical MSI markers, which has been conducted on a small sample, fails to draw unambiguous conclusions, but shows the need to investigate this phenomenon on large samples and by using genetic methods.

[Characteristics of the metabolic status of children of the first year of life with protein-energy deficiency depending on the gestational age at birth.] / Характеристика метаболического статуса детей первого года жизни с белково-энергетической недостаточностью в зависимости от гестационного возраста при рождении.

Comparative analysis of energy-plastic exchange indicators in mature and premature children of the first year of life in the development of protein-energy malnutrition (PEM) was carried out. Unidirectional changes are revealed, including an increase in creatinine, lactate and creatine phosphokinase activity levels, suggesting a n increasing muscle mass deficit against the background of glucose anaerobic oxidation activation. In preterm infants, glucose and triacylglicerine levels decrease, which reflects uncompensated insufficiency of energy substrates and, accordingly, ATP level. Multidirectional deviations in metabolism are pyruvate and ATP content: increase in full-term infants and decrease in preterm infants, that should be taken into account when monitoring condition of children with PEM. A significant decrease of pyruvic acid in preterm infants against the background of the levels of total protein, albumin, hemoglobin, and transferrin, not exceeding reference values, can obviously testify to the active use of this integral metabolite to maintain the fund of substituted amino acids. Development of this pathology in both mature and premature infants creates a pre-morbid background for iron deficiency anemia-diagnostic panel, which should be supplemented by calculation of transferrin saturation coefficient. Regardless of gestational age in childbirth during the formation of PEM, the lipid spectrum is rearranged according to atherogenic type: at normal values of total cholesterol, there is a significant increase in low and very low density lipoproteins with an increase in the atherogenicity coefficient. This singles out children with the pathology in question as a risk group for the development of the atherosclerotic process later, which justifies the recommendation to control the lipid profile in children of the first year of life.

[The first experience of PD-L1 testing of triple negative breast cancer with marker SP142 in Russia]. / Pervyi opyt PD-L1-testirovaniya troinogo negativnogo raka molochnoi zhelezy markerom SP142 v Rossii.

AIM OF STUDY: To evaluate the pecularities of PD-L1 expression in triple negative breast cancer (TNBC) in the Russian population. MATERIALS AND METHODS: For 7 months, within a scientific study of the Russian Society of Clinical Oncology (RUSSCO), we determined the PD-L1 status of 58 patients with TNBC. In each case, an immunohistochemical study was performed in a closed Ventana Bench Mark Ultra automatic stainer using a closed protocol with rabbit monoclonal antibodies Ventana PD-L1 SP142 and Opti View DAB IHC Detection Kit with Opti View Amplification Kit. RESULTS: Positive PD-L1 status in TNBC was detected in 37.93% of cases. Almost all tumors had an expression level of up to 10%. Only 5.17% of cases showed ligand expression on tumor cells. CONCLUSIONS: According to the results of the first experience of testing PD-L1 in TNLM in Russia, it was possible to obtain data comparable to the same data of large international studies. RUSSCO's information and logistic support allows making this analysis available to all citizens of the country.

[Immunohistochemical study of the MSI phenotype of urothelial bladder cancer]. / Issledovanie MSI-fenotipa urotelial'nogo raka mochevogo puzyria immunogistokhimicheskim metodom.

OBJECTIVE: To investigate the MSI phenotype of urothelial bladder cancer (BC) by immunohistochemistry (IHC) and to assess its relationship to the prognostic factors of the disease and PD-L1 status of BC. MATERIAL AND METHODS: Using the surgical and biopsy materials obtained from 50 patients with BC during examination and treatment, the investigators studied the MSI phenotype by IHC. A system was proposed to assess the IHC expression of MSI proteins, by taking into account the intensity of nuclear staining and the area occupied by tumor cells with stained nuclei. RESULTS: An analysis of the results of an IHG study of MSI protein expression revealed a high direct correlation between the nuclear staining intensity of tumor cells and the percentage of tumor area occupied by the latter. The lack/decrease of the expression of the studied proteins was associated with the stage (T) and the presence of a high-grade tumor. The heterogeneous expression of the studied proteins (PMS2, MLH1, and MSH6) was noted to be 10, 30, and 40%, respectively. CONCLUSION: A high direct correlation was observed between the nuclear staining intensity of tumor cells and the percentage of the area occupied by the latter. There was a relationship between the lack and/or decrease of the expression of MSI proteins (mainly PMS2 and MLH1 and to a lesser extent MSH6) and the grade of BC, as well as stage T; there was a tendency to decrease the expression of the studied proteins in the area of invasive tumor growth, which confirms the prognostic role of MSI proteins. The most pronounced heterogeneity of IHC expression was noted with MSH6; the least one was seen with PMS2. There was a predominance of high-grade surface carcinomas (T1) among the heterogeneously stained tumors. In the case of positive PD-L1 status, there was decreased PMS2 and MLH1 expression and pronounced MSH6 expression; no significant relationship was found due to the small number of these cases. The authors consider it necessary to conduct further studies of the relationship between the MSI phenotype and the PD-L1 status of BC.

[Study of the PD-L1 status in breast cancer, by using the SP142 monoclonal antibody, and the prospects for determining treatment policy]. / Issledovanie PD-L1-statusa raka molochnoi zhelezy s ispol'zovaniem monoklonal'nogo antitela SP142 i perspektivy dlia opredeleniia lechebnoi taktiki.

OBJECTIVE: To investigate the expression of programmed death ligand 1 (PD-L1) in triple-negative and luminal B, HER2-negative breast cancer, by using the SP142 antibody and to assess the association of the PD-L1 status with prognosis for patients. MATERIAL AND METHODS: The study was conducted using surgical materials (full sections) obtained from 72 patients. The sections were stained with the SP142 PD-L1 antibody. RESULTS: Differences were found in the detection rates of the PD-L1-positive status in the primary tumor and regional metastasis (primary tumor in 26 (36.1%) cases and metastasis in 18 (47.4%) of the 38 cases). By and large, the PD-L1-positive status was less common in the patients receiving neoadjuvant chemotherapy (25.8% versus 53.7%); however, it should be noted that the PD-L1-positive status was more often detected in those who had a higher residual cancer burden (RCB) (RCB-III versus RCB-II). CONCLUSION: Considering the findings, it is necessary to clarify the status of not only a primary focus, but also clinically significant metastases, especially if the primary tumor has yielded a negative result. The patients with disease progression who receive standard therapy regimens may have a chance for a good result when using PD-1/PD-L1 blockers. At the same time, the association of the PD-L1 status with RCB may affect the choice of adjuvant treatment policy.

Quantification of the diagenesis-designating metals in sediments by ICP-MS: Comparison of different sample preparation methods.

Concentrations of specific metals in sediments are of immense importance in geochemical studies related to diagenetic processes, and quantification can be most reliably performed by ICP-MS after bringing the sample into solution. It is the latter analytical operation that generates a discord as no common sample preparation approach has been yet accepted. To resolve this challenge, we compared different modes of acidic sample digestion (in open and closed systems, applying in the latter case conventional or microwave heating) using a lake sediment reference sample. It was shown that the sample treatment in an autoclave with resistance heating provides the best fit (relative deviation < 7%) to certified values for metals generally used as diagenetic proxies (U, Sm, Nd, Th, La, Sc, Sr, etc.). Furthermore, the thermally convective pressure digestion appears to be more cost-efficient than microwave-assisted digestion (because of lower reagent consumption and cheaper removable PFE liners), an important asset for systematic geochemical mapping. In order to prove the applicability of the developed digestion procedure for different sediments, a marine sample from the East Siberian Sea was analyzed independently in two laboratories and across all representative metals the results acquired were found in good agreement.

[Blood group as a predictor of glycemia and lactatemia in critical condition patients.]

Hyperlactatemia is a dangerous metabolic shift, manifested in an increase in the blood lactate content above 2.2 mmol / l. Changes in the content of lactate affect the function of vital organs and systems, disrupting the course of biochemical processes. This condition accompanies a large number of clinical situations, also it is a major factor, worsening the clinical outcome and the patient's condition. The study of the characteristics of its occurrence and metabolic relationships helps to fulfill the diagnostic criteria for assessing the likelihood of an adverse outcome. The purpose of this study is to assess the variability of the content of lactate and glucose, as well as the glucose/lactate ratio, depending on the group of blood in the ABO system and the nosology of patients. 250 patients with various diseases whose condition required intensive care were examined, the middle age was 61.08 ± 13.6 years, and 446 practically healthy individuals who did not have acute somatic and infectious diseases, the middle age was 26.83 ± 1.46 years . We investigated the content of lactate and glucose, the glucose/lactate ratio, determined the belonging of the blood to ABO system. Statistical processing of the results was performed in the IBM SPSS Statistica 23.0. The data obtained indicate that patients with B (III) blood group have the highest lactate content in venous blood compared to the general population, and patients with AB (IV) blood group have the highest blood glucose level. In the group of patients with the highest median lactate content, the lowest glucose content was observed. The data presented demonstrate the importance of measuring and monitoring glucose, lactate, and their ratio in the management of patients in a life-threatening condition. Evaluation of metabolic variability depending on the different blood grouping of the ABO system in critical conditions involving hyperlactataemia can serve as an important tool in modeling a personalized approach to the patient.

[PD-L1 status in breast cancer]. / PD-L1-status raka molochnoi zhelezy.

OBJECTIVE: To investigate the expression of PD-L1 in triple-negative and luminal B, HER2-negative breast carcinoma and to assess the association of the tumor PD-L1 status with the prognosis of the disease. SUBJECT AND METHODS: The PD-L1-status of primary tumor was studied in 72 patients with breast cancer, by using an immunohistochemical method. RESULTS: Differences were found in the incidence of carcinomas with PD-L1 expression in tumor cells depending on the molecular genetic type: there was a positive PD-L1 status in luminal B, HER2-negative tumors in 4 (14.81%) of 27 cases and in triple-negative tumors in 17 (37.78%) of 45 cases. An analysis of tumors after neoadjuvant therapy revealed a positive PD-L1-status in tumor cells in 1 of 18 patients with a moderate residual tumor load and in 6 of 13 with a high residual tumor load (5.56 and 46.15%, respectively), as estimated by the RCB system. CONCLUSION: The positive PD-L1 status in triple-negative breast cancer was determined more than 2 times as frequently as in luminal B, HER2-negative breast cancer (37.78 and 14.81%). There was a considerable correlation between the high residual tumor load and the positive tumor PD-L1 status.

What are the current analytical approaches for sediment analysis related to the study of diagenesis? Highlights from 2010 to 2018.

This review is aimed at critical analysis of current and emerging capabilities of analytical methods as employed for sediment analysis. An emphasis is given to the most reliable experimental strategies used to acquiring analytical information that is relevant to study the diagenetic processes affecting the composition of sediments. Advanced analytical methodology in use basically rely on the application of mass spectrometry (or a few alternative techniques) to determine various inorganic elements, to measure their isotopic composition, to characterize organic matter, or to identify and quantify its principle components (biomarkers). Also brought into focus are sample preparation techniques which - given the complexity of sediment matrices and the diverse and multiple nature of analytes under scrutiny - are often a key for successful analysis.

[Molecular markers of the oral mucosa damage in patients with leukemia.]

The criteria for early manifestations of mucosal lesions in patients with acute and chronic leukemia are distinguished: in the absence of complaints and changes in the index assessment of the oral cavity, a block of initial dental signs in the form of edema of the tongue and the smoothness of its papillae, pallor of the oral mucosa in patients with leukemia. In theoral fluid of patients with the initial signs of dental pathology in acute leukemia, a specific spectrum of disorders was established in the form of the lowest antibody level of immunoglobulin A class to transglutaminase in combination with a high amount of Ig G antibodies to transglutaminase - 8.73 ± 0.92 U / ml, which maximal level exceeds the reference limits by almost 4 times (38.80 U / ml) - that indicates structural changes in the connective tissues of the oral cavity. Under the influence of chemotherapy in these patients, the inversion of clinical manifestations of dental pathology was noted: stomatitis II (medium) severity is formed, bypassing I degree, accompanied by a 4-fold increase of IgA antibodies to transglutaminase (4.03 ± 0, 77 U / ml, p < 0.05) in the oral fluid of patients with acute leukemia and 2.5 times for chronic leukemias (3.24 ± 0.47 U / ml, p < 0.05) compared to pre-treatment period. In this regard, it is recommended to determine the content in the oral fluid for antibodies to transglutaminase in patients with leukemia for verification of the degree of inflammatory-destructive process and its recovery. The peculiarity of immunological shifts in stomatitis of the 1st degree of severity is the highest level of IgG antibodies to transglutaminase (9.98 ± 1.50 U / ml) in the oral fluid, and at II-III degree the molecular manifestations of damage are smoother, which indicates the depression of immune processes.