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Relevance. The authors of this article present a new method of epiretinal membrane preparation, its’ distinctive feature is formation of ultrathin slices, which allows a more detailed study of the structure of epiretinal and inner limiting membranes removed from the retinal surface during surgery for epiretinal fibrosis. Purpose. To present the ultrastructural features of epiretinal membranes removed during surgery for a more detailed understanding of the pathogenesis and consequences of surgery for idiopathic epiretinal membrane. Material and methods. Three patients (3 eyes) diagnosed with idiopathic epiretinal fibrosis were included in the study. The patients underwent surgical intervention, during which the removed samples of epiretinal and inner limiting membranes were subjected to fixation according to the author’s technique and immunohistochemical staining. Results. The data obtained in conjunction with the surgical technique performed, confirmed that laminin β2 is a characteristic marker of internal limiting membrane. When comparing the data of immunohistochemical study and intraoperative determination of the character of the removed membranes, it was found that the latter method is, in some cases, insufficiently accurate. Conclusion. The results obtained in this study cast doubt on the possibility of preserving the internal limiting membrane in a number of surgical cases for epiretinal fibrosis, and also indicate the inaccuracy of assessing the type of the removed membrane during surgical intervention. Key words: immunohistochemical study, internal limiting membrane, epiretinal membrane

The immunosuppressive status of the tumor microenvironment (TME) remains poorly defined due to a lack of understanding regarding the function of tumor-associated macrophages (TAMs), which are abundant in the TME. TAMs are crucial drivers of tumor progression, metastasis, and resistance to therapy. Intra- and inter-tumoral spatial heterogeneities are potential keys to understanding the relationships between subpopulations of TAMs and their functions. Antitumor M1-like and pro-tumor M2-like TAMs coexist within tumors, and the opposing effects of these M1/M2 subpopulations on tumors directly impact current strategies to improve antitumor immune responses. Recent studies have found significant differences among monocytes or macrophages from distinct tumors, and other investigations have explored the existence of diverse TAM subsets at the molecular level. In this review, we discuss emerging evidence highlighting the redefinition of TAM subpopulations and functions in the TME and the possibility of separating macrophage subsets with distinct functions into antitumor M1-like and pro-tumor M2-like TAMs during the development of tumors. Such redefinition may relate to the differential cellular origin and monocyte and macrophage plasticity or heterogeneity of TAMs, which all potentially impact macrophage biomarkers and our understanding of how the phenotypes of TAMs are dictated by their ontogeny, activation status, and localization. Therefore, the detailed landscape of TAMs must be deciphered with the integration of new technologies, such as multiplexed immunohistochemistry (mIHC), mass cytometry by time-of-flight (CyTOF), single-cell RNA-seq (scRNA-seq), spatial transcriptomics, and systems biology approaches, for analyses of the TME.

An autopsy of a patient in Japan with coronavirus disease indicated pneumonia lung pathology, manifested as diffuse alveolar damage. We detected severe acute respiratory syndrome coronavirus 2 antigen in alveolar epithelial cells and macrophages. Coronavirus disease is essentially a lower respiratory tract disease characterized by direct viral injury of alveolar epithelial cells.

Although radiological diagnostics have been progressing, pathological diagnosis remains the most reliable method for diagnosing liver tumors. In some cases, definite pathological diagnosis cannot be obtained by histological evaluation alone, especially when the sample is a small biopsy; in such cases, immunohistochemical staining is very useful. Immunohistochemistry is the most frequently used technique for molecular pathological diagnosis due to its broad application, ease of performance and evaluation, and reasonable cost. The results occasionally reflect specific genetic mutations. The immunohistochemical markers of hepatocellular carcinoma include those of hepatocellular differentiation—such as hepatocyte paraffin 1 and arginase-1—and those of malignant hepatocytes—such as glypican-3, heat shock protein 70, and glutamine synthetase (GS). To classify the subtypes of hepatocellular adenoma, examination of several immunohistochemical markers, such as liver fatty acid-binding protein, GS, and serum amyloid A, is indispensable. Immunohistochemical staining for GS is also important for the diagnosis of focal nodular hyperplasia. The representative immunohistochemical markers of intrahepatic cholangiocarcinoma include cytokeratin (CK) 7 and CK19. In this article, we provide an overview of the application of immunohistochemistry in the pathological diagnosis of liver tumors referring to the association with genetic alterations. Furthermore, we aimed to explain the practical points in the differential diagnosis of liver tumors by immunohistochemical staining.

Artificial Intelligence (AI) and Deep Learning (DL) hold great promise to transform pathology practice. Currently, the majority of commercially available products and AI research focuses on end-toend AI, i.e., an approach in which the model learns all the steps between the initial input and the final output, providing qualitative or semiquantitative (class) data. A complementary or alternative approach to analyse histomorphology is using DL-based segmentation of relevant histological compartments and cells, followed by extraction of relevant quantitative data (features). If done on a large scale, it is termed pathomics, representing a novel -omics approach for morphology at the microscopical level. Pathomics complements molecular omics, like genomics or transcriptomics, or radiomics, which aims at quantifying radiology images at the macroscopic level. This lecture will explore the potential of pathomics and compare it with end-to-end models, focusing on kidney pathology.

Multiplexed imaging is a powerful approach for studying the spatial organization and cellular composition of intact tissues at single-cell resolution. The last decade has seen a rapid expansion in the development and commercialization of spatial biology techniques. These methods include technologies that probe RNA molecules using imaging-based approaches or spatial barcoding techniques. In addition, proteins may be targeted with antibodies applied to thin sections as well as thick tissue volumes using a variety of approaches. These methods vary in the optical resolution, tissue volume, and number and type of targets (RNA, protein, or both) that can be imaged in a specimen. These technologies have been foundational for the construction of single cell atlases and the study of naturally occurring cancers. Despite their promise, widespread adoption of these methods remains limited by high costs, specialized equipment, and the need for significant technical expertise in tissue processing, reagent validation, image acquisition, and data analysis. To address these barriers, community-driven initiatives led by the Human BioMolecular Atlas Program (HuBMAP) and IBEX Imaging Community are working to streamline and democratize multiplexed imaging. By sharing validated antibody panels, protocols, and best practices, these efforts are reducing the time, resources, and expertise required to generate high-quality spatial data— accelerating discovery through collaboration.

In the last 43 years, thanks to fantastic collaborators, we used antibodies in immunostaining and published a few firsts: TP53 in breast cancer (1988), FFPE-proofs anti Ki-67 antibody MIB 1 and the citrate antigen retrieval buffer (1992), reticular cells in human bone marrow (1993), BCL6 in germinal centers (1995), PRDM1 in plasma cells (2005), the sub cellular localization and distribution of AID and the function of IRF4 (2006). From 2013 to 2021 we dissected the effects of tissue processing on antigenicity, a body of discoveries which led to a hyperplexed (>15 markers) multiplexing method, Multiple Iterative Labeling by Antibody Neodeposition (MILAN)1,2. The application of MILAN and the complexity of data obtained ( 100 markers, millions of single cells) landed us in the rarefied world of dimensionality reduction algorithms, where math rules and statistical significance replaces “representative images”. Because of these latest developments, we discovered that the human eye ability to discriminate shades of grey is very limited (less than 64/256)3, thus low levels of staining are routinely missed and eye-guided assessment is unreliable. Bioinformatics tools we developed (BRAQUE)4 provides highest sensitivity, granularity and robustness based on objective statistical parameters. Biologists and Pathologists need to believe (in math)5 rather than see with their own flawed eyes.

Different parts of Araucaria bidiwillii (bunya pin) trees, such as nuts, seeds, bark, and shoots, are widely used in cooking, tea, and traditional medicines around the world. The shoots essential oil (EO) has not yet been studied. Herein, the chemical profile of A. bidiwillii shoots EO (ABSEO) was created by GC–MS analysis. Additionally, the in vivo oral and topical anti-inflammatory effect against carrageenan-induced models, as well as antipyretic potentiality of ABSEO and its nanoemulsion were evaluated. Forty-three terpenoid components were identified and categorized as mono- (42.94%), sesqui- (31.66%), and diterpenes (23.74%). The main compounds of the ABSEO were beyerene (20.81%), α-pinene (16.21%), D-limonene (14.22%), germacrene D (6.69%), β-humulene (4.14%), and sabinene (4.12%). The ABSEO and its nanoemulsion exhibited significant inflammation suppression in carrageenan-induced rat paw edema model, in both oral (50 and 100 mg/kg) and topical (5% in soyabean oil) routes, compared to the control and reference drugs groups. All the results demonstrated the significant inflammation reduction via the inflammatory cytokines (IL-1β and IL8), nitrosative (NO), and prostaglandin E2 (PGE2) supported by the histopathological studies and immunohistochemical assessment of MMP-9 and NF-κβ levels in paw tissues. Moreover, the oral administration of ABSEO and its nanoemulsion (50 and 100 mg/kg) exhibited antipyretic activity in rats, demonstrated by the inhibition of hyperthermia induced by intramuscular injection of brewer’s yeast. These findings advised that the use of ABSEO and its nanoemulsion against numerous inflammatory and hyperthermia ailments that could be attributed to its active constituents.

SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 4 (SMARCA4)-deficient tumors are rare and highly aggressive tumors characterized by a loss of SMARCA4 expression, and SMARCA4-deficient tumors in the adnexal area of the uterus are particularly rare. The present study describes the case of a 64-year-old woman who was admitted to Weifang People's Hospital (Weifang, China) with abdominal distension, and was observed to have a mass with ascites in the adnexal area of the uterus. Based on clinical, imaging and pathological findings, the patient was diagnosed with a SMARCA4-deficient adnexal tumor with ascites. Biopsy of the left and right adnexal lesions was performed, and the patient was administered chemotherapy. After one cycle of bevacizumab, sindilizumab and carboplatin, no further treatment was administered. After biopsy and chemotherapy, the abdominal distension was alleviated and the general condition of the patient was satisfactory. The patient was followed up and died 3 months after treatment. Notably, it is important to avoid misdiagnosing this tumor as other types of adnexal uterine tumors, and morphological and immunohistochemical features may be useful for diagnosing primary SMARCA4-deficient tumors in the adnexal area of the uterus.