Penile squamous cell carcinoma (pSCC) is a rare cancer associated with a relatively poor prognosis. The WHO classifies pSCC into HPV-associated and HPV-independent subtypes, with HPV-positive pSCCs showing slightly better outcomes. A key negative prognostic marker is TP53 mutation. The aim of this study was to evaluate the concordance of (1) p16 immunohistochemistry with HPV status determined by PCR, (2) p53 immunohistochemistry with TP53 mutational status obtained by NGS, and (3) prognostic impact of overexpression and null phenotype p53 immunohistochemistry profiles. Analyzing invasive pSCC from 209 patients, we assessed concordance between p16 immunohistochemistry and HPV status (n = 132) determined by quantitative PCR, obtaining Cohen's kappa Κ = 0.62 (p < 0.0001). For p53 immunohistochemistry versus TP53 status by NGS (n = 145), Cohen's kappa reached Κ = 0.693 (p < 0.0001). We found worse overall survival (Hazard ratio = 3.33, 95% CIs 1.92-5.56, p < 0.0001) in p53 mutated cases (n = 38, both overexpression and null phenotype) compared to wild type (n = 171), but without a significant difference between both mutated profiles. Both HPV status and p53 profile can be reliably assessed by immunohistochemistry with substantial agreement. Patients with a mutated p53 profile showed significantly worse survival, irrespective of the mutated profile variant.
The diagnosis of T-cell lymphoma is challenging, and establishing clonality is an important aspect of the workup. Molecular testing to establish T-cell clonality though T-cell receptor (TCR) gene rearrangement polymerase chain reaction is the most common modality and can detect clonal rearrangements in ∼90% of T-cell lymphomas, but is limited by increased turnaround time and cost and the need for sufficient material for testing. In the maturation of T cells, the TCR locus rearrangement will result in the mutually exclusive expression of either the TCR beta-chain constant domain 1 or 2 (TRBC1, TRBC2). Antibodies to TRBC1 and TRBC2 have been used in the setting of flow cytometry, and more recently immunohistochemistry, to establish T cell clonality. This study evaluates the utility of a TRBC1/CD3 dual stain for the diagnosis of T-cell lymphoma. The laboratory information system was searched from 2019 through 2025 for cytology samples with a diagnosis of T-cell lymphoma. Twelve samples were identified along with controls. Dual color immunohistochemistry for TRBC1 and CD3 was performed using a brown chromogen for TRBC1 and a red chromogen for CD3. The proportion of cells staining for TRBC1 and CD3 was scored. A monoclonal T-cell population was detected in all samples of T-cell lymphoma. All B-cell lymphomas and benign samples showed a polyclonal staining pattern consisting of a mixture of brown and red cells. Using immunohistochemistry for TRBC1 paired with CD3 is a reliable surrogate for T-cell clonality testing in cytology specimens and can support a diagnosis of T-cell lymphoma on limited material.
Immunohistochemistry (IHC) is a widely used technique for the identification, localization, and analysis of protein expression in tissues, playing a critical role in pathological diagnosis and validation of data derived from molecular biology. Despite its well-established use in mammalian tissues, applying IHC to fish still presents challenges, particularly due to the use of heterologous antibodies and the presence of paralogous genes. In this context, the present study aimed to standardize an immunohistochemistry protocol adapted for fish skeletal muscle using a biotinylated monoclonal antibody designed against a conserved region of the waslb gene for pacu (Piaractus mesopotamicus) and zebrafish (Danio rerio). Muscle samples were fixed in 4% paraformaldehyde, embedded in paraffin, and subjected to heat-induced antigen retrieval. After blocking nonspecific binding sites, histological sections were incubated with the primary antibody at an optimized 1:50 dilution, followed by detection with fluorophore-conjugated streptavidin (1:200) and nuclear counterstaining with DAPI. Reaction specificity was assessed using negative controls with a 20,000-fold molar excess of the immunizing peptide. The standardized protocol resulted in specific and reproducible labeling of the waslb protein in the skeletal muscle of both species, with no signal observed in negative controls; however, it is important to emphasize that this protocol can also be applied to antibodies against other fish genes. Thus, this method represents a reliable tool for protein analysis in fish muscle tissues, contributing to physiological, experimental, and comparative studies in non-mammalian models. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Immunohistochemical reaction against the WASLB antibody in paraffin-embedded skeletal muscle sections of pacus and zebrafish.
Fetal growth restriction (FGR) is frequently defined as the failure of the fetus to reach its genetically predetermined growth potential. Heat shock proteins (HSPs) are extreme-temperature-resistant molecules that help proteostasis. The aim of this prospective case-control immunohistochemistry study is to evaluate the expression of HSP90 and HSP70 in the placentas of pregnancies complicated with FGR and compare their levels with the control placentas of normal-growth pregnancies. A prospective case-control study was conducted including people undergoing singleton pregnancies who gave birth in a tertiary university hospital in Central Greece. Participants were divided into two equal groups: an FGR pregnancy group and a control group with normal growth. Immunohistochemistry of placental samples was assessed using anti-HSP90 alpha/beta antibody (clone F-8, Santa Cruz Biotechnology, Dallas, TX, USA) and anti-HSC70/HSP70 antibody (clone W27, sc-24, Santa Cruz Biotechnology, Dallas, TX, USA). A scoring system was created to quantify the expression of HSP90 and HSP70 in each sample, and the grade of staining was measured at four points. A total of 80 pregnant people were prospectively enrolled in our study, with 40 in each group. Both constitutive (HSP90β and HSC70/HSPA8) and stress-inducible (HSP90α and HSP70/HSPA1A/B) isoforms were analyzed. When comparing the total score of HSP expression, a statistically significant difference was observed for both HSP90 and HSP70. For HSP90 expression, only the Hofbauer cell's stain was identified as a statistically significant independent factor, meaning that its positive expression was observed in Hofbauer cells. For HSP70 expression, only the staining of syncytiotrophoblasts was identified as an independent factor. FGR is a common pregnancy complication and a leading cause of stillbirth, neonatal mortality, and short- and long-term neonatal morbidity worldwide. Based on our findings, the lower expression levels of both HSP90 and HSP70 are associated with FGR, revealing a possible association with stress response in FGR pathophysiology. However, more robust data from larger-scale prospective studies are needed to elucidate the possible role of HSPs as potential FGR biomarkers.
Per National Comprehensive Cancer Network guidelines for acute myeloid leukemia (AML), bone marrow (BM) evaluation is recommended after starting induction chemotherapy at 14 to 21 days (mid-induction) and 30 to 35 days (postinduction). Timely assessment of residual morphologic and molecular disease is challenging at these time points, given the low cellularity, the lack of CD34 or CD117 expression by blasts in most NPM1-mutated (NPM1mut) AML, and the longer turnaround time for molecular testing. NPM1 mut AML undergoing BM evaluation at mid- and postinduction with concurrent flow cytometry and NPM1 real-time polymerase chain reaction or next-generation sequencing was evaluated. Different cell types positive by NPM1 immunohistochemistry (IHC) were enumerated, and the results were compared with flow cytometry and molecular findings. NPM1 IHC demonstrated a sensitivity of 78% and 41% for detecting residual mutation at mid- and postinduction, respectively, with a positive predictive value ranging from 88% to 100%. Of 16 IHC-negative cases, 3 had no residual NPM1 mutation, 12 had very low-level mutation at a variant allele frequency of less than or equal to 21 transcripts, and only 1 had a high-level mutation in a markedly hypocellular BM. Of 24 IHC-positive cases, 22 had residual NPM1 mutation, and 2 showed false-positive staining in only a single cell with no residual NPM1 mutation. The level of IHC staining was mostly proportional to the level of residual mutation. Mutation-specific NPM1 IHC is a valuable tool for a rapid assessment of residual morphologic and molecular disease in mid- and postinduction NPM1mut AML.
To assess the frequency and level of expression of enhancer of zeste homologue 2 in cutaneous squamous cell carcinoma, and compare then with normal skin. The retrospective, descriptive study was conducted at the Department of Morbid Anatomy and Histopathology, University of Health Sciences, Lahore, Pakistan, and comprised data from January 2016 to April 2023 related to cutaneous squamous cell carcinoma in group A and normal skin cases in group B that were evaluated and compared for enhancer of zeste homologue 2 immunohistochemistry expression and positivity. Group A cases with increased immunohistochemistry expression were tested by fluorescence in situ hybridisation for enhancer of zeste homologue 2 amplification. Data was analysed using SPSS 22. Of the 60 patients, 30(50%) were in group A; 19(63.3%) males and 11(36.7%) females with mean age 48±16.5 years. There were 30(50%) patients in group B; 29(96.7%) females and 1(0.3%) male with mean age 47.6±11.3 years. There were 25(83.3%) well-differentiated cases in group A. Enhancer of zeste homologue 2 positivity was noted in 25(83.3%) case in group A compared to 6(20%) in group B (p<0.001). The enhancer of zeste homologue 2 expression level was also significantly higher in group A than group B (p<0.001). Immunohistochemistry overexpression of enhancer of zeste homologue 2 was found in 13(43.3%) group A cases, and, of them, 6(46.15%) showed enhancer of zeste homologue 2 amplification. Enhancer of zeste homologue 2 overexpression was noted in cutaneous squamous cell carcinoma cases, indicating that enhancer of zeste homologue 2 had a potential role in cutaneous squamous cell carcinoma tumourigenesis.
Asthma is a prevalent chronic airway disease with unmet therapeutic needs. Aster tataricus L. f. (AT), a traditional herbal medicine used for respiratory conditions, shows potential for asthma treatment; however, its underlying pharmacological mechanisms remain largely unexplored. This study aimed to systematically investigate the potential therapeutic effects of AT against asthma and elucidate its underlying molecular mechanisms by integrating network pharmacology prediction, molecular docking validation, and in vivo experimental confirmation. To evaluate the anti-asthmatic potential of AT, an Ovalbumin (OVA)-induced murine model of allergic airway inflammation was employed. Pathological alterations in airway inflammation and remodeling were assessed via histochemistry (H&E, PAS, Masson's trichrome). Concurrently, the levels of key cytokines (IL-4, IL-5, IL-10, IL-13, IFN-γ) in Bronchoalveolar Lavage Fluid (BALF) were quantified by Enzyme-Linked Immunosorbent Assay (ELISA). Network pharmacology predictions and molecular docking analyses were integrated to identify potential targets and pathways, which were subsequently validated through Immunohistochemistry (IHC) and Immunofluorescence (IF). In a murine model of allergic airway inflammation, AT treatment significantly alleviated airway inflammation and fibrosis. This therapeutic effect was characterized by a distinct shift in the BALF cytokines: a reduction in the levels of IL-4, IL-5, and IL-13, coupled with an elevation in IL-10 and IFN-γ. This shift was accompanied by diminished inflammatory cell infiltration and collagen deposition in lung tissue. Network pharmacology predicted core targets (e.g., EGFR, STAT3, TLR4, MMP9) and key pathways (e.g., PI3K-Akt and JAK-STAT). Experimental validation confirmed that AT downregulated the expression of these key targets. Furthermore, molecular docking revealed stable binding between active constituents in AT (such as Aster saponin F) and the core targets, suggesting a potential mechanism of action. This study demonstrates the potential anti-asthmatic effects of AT in a murine model of allergic airway inflammation. The integrated research strategy reveals its multi-component synergistic mechanism of action. AT exerts potentially anti-asthmatic effects by means of multi-component and multi-target mechanisms, with the regulation of the PI3K-Akt and JAK-STAT pathways. This study elucidates the anti-asthmatic mechanism of AT and provides scientific evidence supporting its clinical application for asthma treatment.
Toxoplasma gondii is a globally distributed intracellular parasitic protozoan. It infects nearly all warm-blooded animals and causes a zoonotic disease of worldwide significance. Currently, the only commercially available vaccine, Toxovax®, is solely used for the prevention of Toxoplasma-induced abortion in sheep, but it has limitations such as a short shelf life and the potential of reversion to virulence. This study evaluated the safety and immune-protective efficacy of two live-attenuated strains RHΔtkl1 and PruΔpp2a-c in sheep. Sheep were immunized via intramuscular injection in the neck with 1 × 107 tachyzoites of RHΔtkl1 or PruΔpp2a-c. Sheep were challenged orally with 5 × 105 type II Pru oocysts at 28 days post-vaccination (dpv), followed by a second challenge on day 70 with 1 × 107 type II Pru tachyzoites injected intramuscularly at 70 dpv. Safety and immuno-protection were evaluated by monitoring clinical symptoms and body temperatures, T. gondii-specific IgG antibody levels, histopathological changes, immunohistochemistry, brain cysts, parasite load, and mouse bioassay results. The results demonstrated that both knockout strains induced only transient fever. Following immunization and subsequent challenge with Pru oocysts, the T. gondii-specific IgG antibody levels in sheep increased rapidly and remained elevated for an extended period. Histopathological analysis indicated mild organ lesions in heart, liver and lung tissues among immunized infected sheep, whereas non-immunized infected sheep exhibited severe widespread inflammation. Immunohistochemical analysis of brain tissue revealed significantly lower values for four parameters (positive cell ratio, density, histochemistry score, immunoreactive score) in immunized groups (P < 0.01). A significant reduction in brain cysts was observed in immunized and challenged sheep (P < 0.01) compared with unimmunized and challenged sheep. The parasite burden of T. gondii in heart tissue was significantly reduced (P < 0.01). Compared with mice inoculated with sheep brain tissue from unimmunized groups challenged with T. gondii Pru oocysts and tachyzoites, mouse bioassay results showed that the mice inoculated with sheep brain tissue from groups immunized with RHΔtkl1 or PruΔpp2a-c tachyzoites and subsequently challenged with T. gondii Pru oocysts and tachyzoites exhibited a significantly lower proportion of positive genomic T. gondii DNA in the brain (P < 0.001), as well as significantly reduced levels of T. gondii-specific antibody IgG in the serum (P < 0.0001). Similarly, mice inoculated with sheep visceral tissue from the same immunized and challenged groups also showed a significantly reduced proportion of positive genomic T. gondii DNA in the brain (P < 0.0001) and significantly reduced levels of T. gondii-specific antibody IgG in the serum (P < 0.0001). The gene knockout strains RHΔtkl1 and PruΔpp2a-c showed a certain degree of safety in sheep, and they induced strong humoral and cellular immune responses in sheep, significantly mitigating acute infection symptoms and tissue damage. Notably, PruΔpp2a-c showed greater potential in suppressing cyst formation. Both strains are potential attenuated candidates against sheep toxoplasmosis.
Postmortem human brains are important research resources to study the mechanisms underlying cerebrovascular features in various neurodegenerative disorders. Immunohistochemical and histochemical staining have been used to examine human brains fixed in neutral-buffered formalin (NBF) for months, years, or decades. Previously, we found that prolonged NBF fixation resulted in differential effects on immunohistochemistry and histochemistry staining on postmortem brains. Here, we further examined the effects of prolonged fixation (1-, 5-, 10-, 15-, 20-years) on stains of known biomarkers of cerebrovascular diseases in the human prefrontal cortex. We included microvasculature markers of the blood vessel wall (Anti-Collagen-IV and Claudin-5), a type III intermediate filament marker (Anti-Vimentin), an activated microglia marker (Anti-CD68), a biomarker for proteolipid protein (Anti-PLP) of oligodendrocytes and a marker for iron accumulation (Anti-Ferritin). We also included Masson's Trichrome Stain (MTS) and Bielschowsky silver stain (BSS). We found that staining intensities of Ferritin, Vimentin, Collagen-IV and BSS decreased with prolonged fixation. No significant differences were observed in the staining intensity of other markers. We therefore recommend performing IHC and HC staining for human brains with the same fixation times to offset any impact on downstream neuropathological analyses, as well as adding the fixation duration as a covariate in the analysis.
Vascular cognitive impairment (VCI) shares major risk factors with heart failure with preserved ejection fraction (HFpEF), including obesity, diabetes and hypertension. Yet VCI research often relies on single-stimulus models, whereas patients experience combined risk factors. We therefore assessed cerebrovascular and cognitive phenotypes in an HFpEF model and investigated underlying mechanisms.Male Lean and Obese ZSF1 rats underwent longitudinal assessments of blood pressure, glucose, cardiac function, and behavioural performance. Cerebral blood flow and neurovascular coupling were assessed by laser speckle contrast imaging. White matter integrity, blood-brain barrier (BBB) permeability, and vascular density were analysed by (immuno)histochemistry. Cortical microvessels were isolated for transcriptomic profiling, and selected targets were validated using multiplex in-situ hybridization.Obese rats exhibited neurovascular uncoupling and impaired short- and long-term memory and spatial learning, accompanied by brain atrophy and reduced myelin. BBB permeability increased at 22-23 weeks and vascular density at 34-35 weeks in Obese vs Lean rats. Transcriptomic analysis of brain microvessels revealed altered processes related to angiogenesis, vasoreactivity, immune mechanisms and vascular remodelling, with consistent downregulation of Trpv4 and Klf2.Obese ZSF1 rats develop progressive neurovascular dysfunction associated with HFpEF onset and reduced Trpv4 and Klf2 expression in cerebral microvessels, two key vasoprotective genes.
The Class II myosin light chain (myl) genes in Chinese perch (Siniperca chuatsi) have not yet been systematically characterised, and relationships with muscle fibre specification, development, and injury-associated remodelling remain unclear. In this study, fast and slow muscle fibres were initially distinguished using myofibrillar ATPase histochemistry. Subsequently, genome-wide mining identified 16 Class II myl genes, comprising eight essential and eight regulatory light-chain subunits. Their conserved-domain features, chromosomal distribution, phylogenetic relationships and expression profiles were analysed. Transcriptomic profiling showed that summed myl transcript abundance was higher in fast muscle than in slow muscle, accounting for 67.2% of the pooled myl transcript pool across the two muscle types (paired t-test, raw P = 0.036). mylpfa, myl1 and mylz3 were the major fast-muscle-associated genes, whereas myl10, myl2b and myl13 were preferentially expressed in slow muscle at the transcript level. These patterns support these genes as candidate fibre-type-associated expression markers. Developmental profiling identified stage-associated myl expression patterns, including a possible expression shift between mylpfb and mylpfa. In the descriptive injury-repair time course (d0-d7), FPKM profiles indicated that fast-muscle-associated genes (mylpfa, mylz3 and myl1) were lower at d1 and recovered by d3, whereas several slow-muscle-associated genes showed biphasic transcript-level increases. The slow-muscle-associated RLC gene mylpfb showed a delayed expression peak at d7. Notably, the embryonic isoform myl6l showed a modest increase from approximately 2 FPKM at d0 to 4-5 FPKM after injury, suggesting a possible injury-associated expression pattern that requires further validation. Together, these findings provide a genome-wide description of the Chinese perch myl gene family and identify candidate fibre-type-associated genes and descriptive injury-associated isoform expression patterns.
Given higher compatible rates between oral lichen planus (OLP) and oral lichenoid lesions (OLLs) in histopathological and clinical features, this study aims to delineate a boundary between equivocal OLP and OLLs by biomarkers. The updated OLP diagnostic criteria in 2016 was our guideline in defining study cases of typical OLP and typical OLL with triggers, which include topical offending agents (OLL-agent), dental restorations (OLL-dental), and systemic offending drugs (OLL-drug). The expression intensity and distribution patterns of CD4, CD8, and CGRP in four groups were detected by immunohistochemistry assay (IHC). A total of 79 cases including OLP (24), OLL-agent (15), OLL-dental (21), and OLL-drug (19) were collected from an oral biopsy laboratory. Band-like distribution patterns of CD4 (100%, score 3), CD8 (54.17%, score 2), and CGRP (87.5%, score 3) in the subepithelial regions of the OLP group significantly differ from the OLL groups (each comparison pair, p = 0.0001). The sensitivity of CD4 (100%), specificity of CD4 (83.64%), negative predictive value of CD4 (100%), and accuracy of CD4 (83.80%) in the OLP group provide results for the diagnostic test evaluation. The band-like distribution pattern of CD4 in the subepithelial region may determine OLP when the biopsy specimen does not show typical microscopic features.
Nephronectin (Npnt) is an extracellular protein with various functions during development and homeostasis of various tissues, but its role during early mouse cornea development remains unexplored. We characterized the expression of Npnt and investigated its function during cornea development. We performed in situ hybridization and immunohistochemistry on ocular sections obtained from embryonic day (E)11.5-E16.5 mouse embryos. Wild-type and Npnt mutant embryos were analyzed for corneal thickness, cell number, and cell proliferation. We assessed the expression of Npnt receptors, Integrin alpha 8 (Itgα8) and Epidermal growth factor receptor (EGFR). We performed in vitro migration assays using periocular mesenchyme (POM) explants in the presence of EGFR and STAT3 inhibitors. Npnt mRNA was expressed in the ectoderm, lens, optic cup, and subsequently in the cornea endothelium. Npnt protein correlated with mRNA staining with additional localization in the epithelial basement membrane and POM. Npnt mutants exhibited reduced corneal thickness and cell numbers compared to wild-type littermates, but there were no differences in cell proliferation. Itgα8 mRNA was diffusely expressed in the POM, but not in the migratory regions. EGFR mRNA was expressed in the region of migratory POM. Robust cell migration occurred from POM explants cultured on either Npnt or Fibronectin (Fn) substrates, but the EGFR pathway inhibitors only attenuated migration on the Npnt substrate. Npnt is expressed in the periocular region during ocular development and contributes to POM migration into the nascent cornea via a mechanism that involves activation of the EGFR signaling pathway.
High-grade endometrial stromal sarcoma (HGESS) is a rare tumor that typically occurs over a wide age range, from 14 to 71 yr, often presenting with nonspecific clinical symptoms. This tumor group displays heterogeneous tumor morphology and a growing number of molecular alterations. The most common alterations are YWHAE::NUTM2 fusion and BCOR gene alterations, including fusions and internal tandem duplications. Here, we report a HGESS with a novel ING3::BCOR fusion. A 74-yr-old female presented with vaginal bleeding and underwent myomectomy. Hematoxylin & eosin (H&E) sections revealed a spindle cell proliferation with both hypocellular and hypercellular areas. Cytologic atypia was mild to moderate, with a mitotic count of 5/10 high-power fields; the background stroma had focal myxoid and collagenous changes. Focal necrosis and pleomorphism were observed. Ancillary studies showed CD10 and diffuse cyclin D1 positivity by immunohistochemistry (IHC). Estrogen receptor (ER), progesterone receptor (PR), ALK1​, S100​, HMB45,​ and CD34​ were negative and p53 showed a wild-type staining pattern​. RNA-based next-generation sequencing showed an ING3::BCOR fusion, confirming the diagnosis of HGESS with ING3::BCOR fusion. HGESS is an evolving entity with an increasing number of genetic alterations, including genetic fusion involving the BCOR gene. HGESS with BCOR fusion should be considered in tumors with a combination of spindle cell growth, myxoid background, foci of coagulative necrosis, diffuse cyclin D1 positivity, and a negative hormone receptor status. Accurate diagnosis requires an integrated approach that combines histopathologic evaluation, IHC, and molecular testing.
Scleromyxedema is a rare, chronic cutaneous mucinosis defined histopathologically by a triad of dermal mucin deposition, fibroblast proliferation, and dermal fibrosis. However, additional underrecognized histopathologic patterns or findings may complicate diagnosis and lead to misclassification. A retrospective review of 22 biopsy-confirmed cases of scleromyxedema was performed to characterize both classic and uncommon histopathologic variants of scleromyxedema, with emphasis on patterns that may obscure or mimic other dermatologic entities. All cases were evaluated for mucin, fibrosis, fibroblast proliferation, inflammatory infiltrates, granulomas, eosinophilic infiltration, and eccrine gland hyperplasia using hematoxylin and eosin and special stains, with immunohistochemistry as it was needed. Sixteen cases (72.7%) demonstrated the classic histologic triad. In contrast, 6 cases (27.3%) showed a granulomatous pattern characterized by interstitial collections of CD68+ histiocytes, multinucleated giant cells, sparse lymphocytes, and mucin, but lacked the full classical triad-posing a risk of misdiagnosis as granuloma annulare or other granulomatous dermatoses. Eosinophilic infiltrates were identified in 4 cases (18.2%), and eccrine gland hyperplasia in 2 cases (9.1%). Both findings coexisted with the classical triad and may mimic inflammatory or adnexal conditions. Scleromyxedema presents with broader histopathologic variability than classically recognized. The granulomatous pattern, which may lack the defining triad, can obscure diagnosis without clinical correlation. In contrast, eosinophilic infiltration and eccrine gland hyperplasia, though infrequent, typically coexist with classic features and should be recognized as part of the histologic spectrum. Awareness of these variants is essential to avoid misdiagnosis and ensure accurate clinicopathologic interpretation.
Prostate cancer (PCa), the second most common cancer, is still a major cause of morbidity and mortality among men worldwide. A particularly aggressive group of PCa, where prognostic biomarkers are still needed, are clinically defined high-risk PCa. We had previously shown that the survival and proliferation factor progranulin (GP88) is a prognostic marker for primary PCa. We aimed in this study to characterize GP88 protein expression in high-risk PCa by immunohistochemistry and to examine its association with prognosis. Immunohistochemical staining for GP88 was performed with TMA of samples from 94 high-risk PCa patients using an H-score. GP88 staining was in a range of 3.69 to 252.51 (median: 109.28). The association of GP88 staining with prognosis was examined by survival analyses (Kaplan-Meier, multivariate Cox's regression analysis). Elevated GP88 expression was associated with a shorter clinical progression-free survival (CPFS) in all PCa patients (P = 0.043), and in the following patient subgroups: Elder PCa patients (> 67 years; P = 0.020), patients with pT3 tumors (P = 0.008), with Gleason scores GS5 and GS6 (P = 0.033 and P = 0.030), and patients without radiation therapy (P = 0.009) at considering that only four patients received an adjuvant radiation therapy. In a multivariate Cox's regression analysis, increased GP88 protein expression (RR = 2.98; P = 0.049) and the preoperative PSA level (RR = 5.29; P = 0.030) appeared as independent prognostic factors for clinical progression. Interestingly, lower GP88 staining in corresponding low Gleason lesions was correlated with the presence of immune cells, suggesting an immune cell suppressive effect of GP88. Altogether, Progranulin (GP88) protein positivity appears very likely to be an independent prognostic factor for clinical progression in high-risk PCa patients.
Stenopodidea represents one of the basal lineages within Pleocyemata, yet the male reproductive system (MRS) of this group remains poorly understood, with limited information available regarding its morphology and function. This study provides the first detailed description of the MRS in four stenopodidean shrimp species from two families: Stenopodidae (Stenopus hispidus, S. scutellatus, and S. spinosus) and Spongicolidae (Microprosthema semilaeve). We analyzed the anatomy, histology, and histochemistry of the testes and vas deferens (VD), as well as spermatozoal ultrastructure, and compared these findings with data from more derived pleocyematans to identify potentially ancestral reproductive traits. Our analyses revealed two principal types of secretion in the VD of Stenopus species and three in M. semilaeve, which together form the presumptive spermatophore. Some secretions occur in small amounts and are restricted to specific VD regions. The layers surrounding the spermatozoa are relatively simple, consisting primarily of a sperm cord enclosed by thin secretory layers, suggesting a plesiomorphic reproductive condition in Stenopodidea. Spermatozoa are elliptical and characterized by a large nucleus in direct contact with the cytoplasm, numerous peripheral vesicles, and a large vesicle containing concentric membrane whorls. This structure, previously described as a lamellar body, is reinterpreted here as a putative acrosome vesicle and differs markedly from acrosome vesicles described in other Pleocyemata. Taken together, the comparatively simple spermatophore architecture and distinctive spermatozoa ultrastructure highlight Stenopodidea as an important lineage for understanding the early evolution of reproductive traits in Pleocyemata.
Autoimmune gastritis (AIG) is characterized by well-described histologic features, yet diagnostic thresholds and reporting practices vary in routine pathology practice. This study aimed to characterize variability in evaluation, ancillary testing, and reporting of AIG across diverse pathology practice settings and training backgrounds. We conducted a national, anonymized, cross-sectional online survey for practicing pathologists who routinely sign out gastric biopsies. The survey assessed diagnostic criteria for AIG, use of ancillary studies, reporting practices, and approaches to overlapping Helicobacter pylori gastritis. Associations between respondent characteristics and diagnostic behaviors were evaluated using χ2 tests and multivariable logistic regression. A total of 163 pathologists completed the survey. Diagnostic thresholds for AIG varied widely, ranging from strict criteria requiring oxyntic gland atrophy, intestinal metaplasia, and enterochromaffin-like-cell hyperplasia to broader approaches recognizing atrophy alone. Only 22% of respondents reported AIG in the main report, while 43% reported in the comments and only one-third routinely subtyped intestinal metaplasia. In all, 52% of respondents routinely ordered gastrin immunohistochemistry in random gastric biopsies. Academic settings were associated with greater reporting detail, while gastrointestinal pathology fellowship training was associated with the use of qualifying terminology such as "early" or "suspicious" AIG (P = .01). Approaches to cases with concurrent H pylori gastritis and oxyntic gland atrophy also showed substantial variability. Diagnostic evaluation and reporting of AIG remain highly variable and are associated with subspecialty training, practice environment, and years of experience. These findings highlight the need for consensus-based guidance to improve consistency, communication, and clinical integration of AIG diagnoses.
To investigate the expression patterns of ERG, PTEN, and c-Myc proteins in prostate adenocarcinoma; analyze the status of the MYC gene; and evaluate their correlations with clinicopathologic parameters and patient prognosis. In this retrospective study, 152 prostate adenocarcinoma cases were analyzed. Immunohistochemistry was performed for ERG, PTEN, and c-Myc protein expression. Fluorescence in situ hybridization was used to assess ERG rearrangement, MYC gene breakage, and amplification. Statistical analyses were conducted to evaluate associations between molecular markers and clinicopathologic variables, as well as their impact on survival outcomes. The positive expression rates for ERG, PTEN, and c-Myc high expression (immunoreactive score ≥5) were 9.9%, 86.8%, and 50%, respectively. The detection rate of MYC gene amplification was 27.6%. Both c-Myc high expression and MYC gene amplification were significantly associated with the high-grade group (defined as Grade Group ≥3 and/or with cribriform pattern), lymphovascular invasion, advanced T stage, metastasis, and poor prognosis, with gene amplification demonstrating higher prognostic specificity. ERG protein expression was significantly correlated with PTEN positivity, whereas no significant association was found between MYC gene amplification and ERG protein expression; only a nonsignificant mutually exclusive trend was observed. c-Myc activation, particularly MYC gene amplification, is a key driver and specific biomarker for an aggressive phenotype and poor prognosis in prostate adenocarcinoma. Combined detection of ERG, PTEN, and c-Myc status contributes to refining risk stratification systems. The lack of significant association between MYC amplification and ERG expression suggests they may represent distinct oncogenic pathways, warranting further validation in large prospective studies.
HER2 is a clinically significant predictive and prognostic marker in a number of epithelial malignant neoplasms. With the introduction of HER2-targeted therapeutic approaches, including antibody-drug conjugates, the need for standardized assessment of HER2 status in tumors of various localizations has become increasingly important. To assess the prevalence of HER2-positive tumors (IHC 3+) in epithelial malignant neoplasms of various localizations in the Russian population using a unified diagnostic approach. This multicenter study included 1.511 tumor tissue samples from patients with epithelial malignant neoplasms of various localizations, obtained from reference pathology centers of the Russian Federation. HER2 status was determined by immunohistochemistry using anti-HER2 antibody (clone 4B5, Ventana) and standardized interpretation criteria developed for gastric cancer and gastroesophageal junction cancer. In cases of equivocal HER2 status (IHC 2+), in situ hybridization was recommended. HER2-positive tumors (IHC 3+) were identified in the majority of the analyzed localizations, with prevalence varying widely depending on tumor type. The highest frequency of HER2 positivity was observed in urothelial carcinoma of the urinary bladder, as well as in gastric and gastroesophageal junction cancers. No HER2-positive cases were detected in lung adenocarcinoma or pancreatic cancer. The study results confirm the reproducibility of HER2 status assessment using a unified diagnostic approach in a multicenter setting. The application of standardized HER2 interpretation criteria ensures data comparability and provides a methodological basis for further clinicopathological studies and for the implementation of HER2-targeted therapeutic strategies in clinical practice. HER2 является клинически значимым предиктивным и прогностическим маркером при ряде эпителиальных злокачественных новообразований. В условиях внедрения HER2-ориентированных терапевтических подходов, включая антитело-лекарственные конъюгаты, возрастает необходимость стандартизированной оценки HER2-статуса при опухолях различных локализаций. Оценить распространенность HER2-позитивных опухолей (ИГХ 3+) при эпителиальных злокачественных новообразованиях различных локализаций в российской популяции при использовании унифицированного диагностического подхода. В многоцентровое исследование включено 1511 образцов опухолевой ткани пациентов с эпителиальными злокачественными новообразованиями различных локализаций, полученных в референсных патолого-анатомических центрах Российской Федерации. HER2-статус определяли методом иммуногистохимии с использованием антитела к HER2 (клон 4B5, Ventana) и стандартизированных критериев интерпретации, разработанных для рака желудка и гастроэзофагального перехода. В случаях неопределенного HER2-статуса (ИГХ 2+) рекомендовалось проведение гибридизации in situ. HER2-позитивные опухоли (ИГХ 3+) выявлялись при большинстве исследованных локализаций, при этом их распространенность варьировала в широких пределах в зависимости от нозологии. Наиболее высокая частота HER2-позитивности отмечена при раке мочевого пузыря, а также при раке желудка и гастроэзофагального перехода. При аденокарциноме легкого и раке поджелудочной железы HER2-позитивные случаи не выявлены. Результаты исследования подтверждают воспроизводимость оценки HER2-статуса при использовании унифицированного диагностического подхода в условиях многоцентрового анализа. Применение стандартизированных критериев интерпретации HER2-статуса обеспечивает сопоставимость данных и создает методологическую основу для дальнейших клинико-патологических исследований и внедрения HER2-ориентированных терапевтических стратегий.