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This article defines immunophenotypes of stromal inflammatory and endothelial cells and fibroblasts 3 months after augmentation of the peri-implant soft tissue using a porcine cross-linked collagen matrix (VCMX). Peri-implant soft tissue samples were obtained from 12 patients at the lining mucosa (LM)-masticatory mucosa (MM) junction before (1) and 3 months after (2) augmentation. Immu-nohistochemical stains were performed to identify inflammatory cells (T [CD3] and B [CD20] lymphocytes, plasma cells [CD138]), macrophages (CD68-proinflammatory, CD163-anti-inflammatory/ reparative), endothelial cells (CD31, CD34), and fibroblasts (CD90, TE-7). Differences in the mean positively stained cells pre- and postaugmentation were analyzed by Wilcoxon signed-rank test. CD31+ endothelial cells showed increased mean numbers in MM2 compared to MM1 (P = .025) and in LM2 compared to LM1 (P = .047). CD163+ anti-inflammatory macrophages showed higher mean numbers in MM2 than in MM1 (P = .021) and in LM2 than in LM1 (P = .012). All other cell phenotypes showed nonsignificant changes between pre- and postaugmentation. This molecular study provides novel insight on the frequency of stromal cell phenotypes in the wound healing process at 3 months postaugmentation with VCMX, with anti-inflammatory CD163+ macrophages being predominant. This should be further investigated to help find novel therapeutic approaches to modulate and promote the VCMX-related healing process. [ABSTRACT FROM AUTHOR]

Pre-determined anatomical locations in the oral cavity were biopsied, and their histomorphology was characterised using haematoxylin and eosin staining (H&E). The most abundant cell type was of dendritic morphology. Lymphocyte foci were not evident in the palatoglossal folds or the gingiva. Immunohistochemical staining (IHC) for validated leukocyte markers followed, including CD3, CD20, CD79α, CD204, and Iba1. Consistent with H&E findings, CD204 immunoreactivity predominated amongst all niches. With the exception of the alveolar mucosa and palatoglossal folds, we also demonstrate a significant difference in the population of macrophages by region for only the Iba1 antigen (p

A free-ranging, adult female two-toed sloth (Choloepus hoffmanni) was brought to a wildlife rescue center in Costa Rica with ocular and auricular myiasis and numerous skin lesions. After one month of unsuccessful systemic and topical antimicrobial treatment, the patient died. A postmortem examination was performed, and tissues were examined histologically, confirming disseminated amebic infection with intralesional trophozoites and cysts in the lungs, liver, eye, heart, spleen, and stomach. Immunohistochemistry identified the ameba as Acanthamoeba sp. A multiplex real-time PCR assay, 18S ribosomal DNA PCR, and sequencing performed on formalin-fixed, paraffin-embedded lung tissue confirmed the Acanthamoeba T17 genotype. The Acanthamoeba genus is in the group of free-living amebas that cause infection in humans and animals, and it is ubiquitous in the environment. Acanthamoeba T17 has been isolated from water and soil, but to our knowledge, this genotype has not been implicated in infections of animals previously and has not been reported from Costa Rica. Systemic Acanthamoeba infection has not been described in sloths previously. We provide a comprehensive literature review describing infections by free-living amebas of the genus Acanthamoeba spp., Balamuthia spp., and Naegleria spp. in domestic, zoo, and wild mammals.

Purpose: To evaluate the effect of interleukin-6 (IL-6) inhibitor (tocilizumab) on bacterial infection-associated bone resorption around implants during osseointegration in rabbits. Materials and Methods: At total of 24 male, 9-monthold New Zealand white rabbits were included, and their two mandibular anterior teeth were extracted. Three months after extraction, 24 one-piece Dentium implants (Ø 2.5 mm, intraosseous length of 12 mm) were inserted in the anterior mandible, and the rabbits were divided into four groups (n = 6 per group). Different treatment methods were used in each group: blank control group (BC); only silk ligation (negative control [NC]); silk ligation and injection with minocycline hydrochloride ointment (positive control [PC]); and silk ligation and injection with tocilizumab at 8 mg/kg via the auricle vein (experimental [EP]). Eight weeks later, the animals were sacrificed, and samples were collected and then analyzed using microcomputed tomography (microCT) scanning, immunohistochemical analysis, and histologic analysis. Results: From the microCT measurement, the ratio of the bone volume to the total volume (BV/TV) in the EP group was 67.00% ± 2.72%, which was higher than that in the other three groups (58.85% ± 2.43% in the BC group, 55.72% ± 2.48% in the PC group, and 36.52% ± 3.02% in the NC group). From immunohistochemical analysis, the expression of IL-6 was found to be higher in the NC group than in the BC, PC, and EP groups, but there was no statistical difference between these three groups. Furthermore, the RANKL (receptor activator of nuclear factor-κB ligand) expression was the lowest in the EP group, followed by the BC group, the PC group, and the NC group, which had the highest expression; there was no difference between the NC and PC groups. Upon histologic analysis, significant new bone was found on the implant surfaces in the EP group, sparse and less new bone could be seen in the BC and PC groups, and the most serious bone resorption occurred in the NC group. Conclusions: Tocilizumab, an inhibitor of IL-6, has a certain effect in preventing bone loss around implants caused by bacterial infection during the osseointegration period. [ABSTRACT FROM AUTHOR]

Secretory carcinoma is a malignant salivary gland tumor, which typically presents as an indolent painless mass within the parotid gland. Involvement of the minor gland is reported but less common. Secretory carcinoma was often misclassified as other salivary gland mimics, particularly acinic cell carcinoma, prior to 2010. It was first recognized as a molecularly distinct salivary gland tumor harboring the same fusion gene as well as histologic and cytogenetic features seen in juvenile breast cancer. Secretory carcinoma is generally managed in the same way as other low-grade salivary gland neoplasms and has a favorable prognosis; however, high-grade transformation requiring aggressive therapeutic interventions have been documented. Recent studies of biologic agents targeting products of this fusion gene offer the promise of a novel therapeutic option for treatment of this malignancy. Due to the limited number of reported cases, the spectrum of clinical behavior, best practices for management, and long-term treatment outcomes for secretory carcinoma remain unclear. A long-standing secretory carcinoma involving minor salivary glands of the mucobuccal fold, which was detected years after it was first noted by the patient, is reported. This case brings to light the importance of a thorough clinical exam during dental visits and reviews diagnostic differentiation of this malignancy from other mimics and discusses decision making for its management. [ABSTRACT FROM AUTHOR]

OBJECTIVES: To determine the role of keratin 17 (K17) as a predictive biomarker for response to chemotherapy by defining thresholds of K17 expression based on immunohistochemical tests that could be used to optimize therapeutic intervention for patients with pancreatic ductal adenocarcinoma (PDAC). METHODS: We profiled K17 expression, a hallmark of the basal molecular subtype of PDAC, by immunohistochemistry in 2 cohorts of formalin-fixed, paraffin-embedded PDACs (n = 305). We determined a K17 threshold of expression to optimize prognostic stratification according to the lowest Akaike information criterion and explored the potential relationship between K17 and chemoresistance by multivariate predictive analyses. RESULTS: Patients with advanced-stage, low K17 PDACs treated using 5-fluorouracil (5-FU)-based chemotherapeutic regimens had 3-fold longer survival than corresponding cases treated with gemcitabine-based chemotherapy. By contrast, PDACs with high K17 did not respond to either regimen. The predictive value of K17 was independent of tumor mutation status and other clinicopathologic variables. CONCLUSIONS: The detection of K17 in 10% or greater of PDAC cells identified patients with shortest survival. Among patients with low K17 PDACs, 5-FU-based treatment was more likely than gemcitabine-based therapies to extend survival.

Ewing sarcoma (ES) is an uncommon mesenchymal neoplasm that typically develops as a bone mass, although up to 30% arise in extraskeletal sites. ES of the gastrointestinal (GI) and hepatobiliary tract is rare and may be misdiagnosed as other, more common neoplasms that occur in these sites. However, the correct classification of extraskeletal ES is important for timely clinical management and prognostication. We reviewed our experience of ES in the GI and hepatobiliary tract in order to further highlight the clinicopathologic features of these neoplasms and document the potential for misdiagnosis in this setting. The archives and consultation files of 6 academic institutions were retrospectively queried for cases of ES occurring in the GI and hepatobiliary tract. The histologic slides and ancillary studies were reviewed and clinical data were retrieved for each case through the electronic medical records, when available. Twenty-three patients with ES in the GI and/or hepatobiliary tract were identified from 2000 to 2022. Of these, 11 were women and 12 were men with a median age of 38 years (range, 2 to 64). Tumor locations included the pancreas (n=5), liver (n=2), stomach (n=3), colorectum (n=3), and small intestine (n=5), as well as tumors involving multiple organs, pelvis and retroperitoneum (n=5). Tumor size varied between 2 cm and 18 cm. Twenty were primary and 3 were metastases. Of the 23 cases, only 17% were initially diagnosed as ES. The most common misdiagnoses involved various forms of neuroendocrine neoplasia due to expression of synaptophysin and other neuroendocrine markers (22%). A wide variety of diagnoses including GI stromal tumor was considered due to aberrant CD117 expression (4%). The diagnosis of ES was ultimately confirmed by detection of the EWSR1 rearrangement in 22 cases. The remaining case was diagnosed using traditional immunohistochemistry. Follow-up information was available in 20 cases, with follow-up time varying between 2 and 256 months. Six patients with follow-up died of disease between 6 and 60 months following initial presentation. Our data indicate ES in the GI and hepatobiliary tract is commonly misdiagnosed leading to a delay in therapy. In light of the attendant therapeutic and prognostic implications, ES should be considered in the differential diagnosis of any GI or hepatobiliary tumor with epithelioid and/or small round cell morphology.

Mannheimia haemolytica-associated abomasitis has been clinically described as a cause of sudden death in lambs, but it is poorly characterized. We describe the pathological features of a severe fibrinonecrotizing abomasitis in 3 lambs that died suddenly. All 3 abomasums had a thickened submucosa due to edema and necrotic areas delimited by bands of degenerate neutrophils with slender nuclei (oat cells) and angiocentric distributions. The overlying mucosa was congested. Myriads of gram-negative coccobacilli were observed within the oat cell bands. M. haemolytica was isolated from the abomasum in all 3 animals and was serotyped as A2 in one of them. Pericarditis and pleuritis were observed in 2 of the lambs. Clostridium spp. were isolated in 1 lamb and detected by immunohistochemistry in the 3 animals, suggesting clostridial co-infection. M. haemolytica should be considered among the differential diagnoses of necrotizing abomasitis in lambs.

Immunohistochemistry (IHC) is used to guide treatment decisions in multiple cancer types. For treatment with checkpoint inhibitors, programmed death ligand 1 (PD-L1) IHC is used as a companion diagnostic. However, the scoring of PD-L1 is complicated by its expression in cancer and immune cells. Separation of cancer and noncancer regions is needed to calculate tumor proportion scores (TPS) of PD-L1, which is based on the percentage of PD-L1-positive cancer cells. Evaluation of PD-L1 expression requires highly experienced pathologists and is often challenging and time-consuming. Here, we used a multi-institutional cohort of 77 lung cancer cases stained centrally with the PD-L1 22C3 clone. We developed a 4-step pipeline for measuring TPS that includes the coregistration of hematoxylin and eosin, PD-L1, and negative control (NC) digital slides for exclusion of necrosis, segmentation of cancer regions, and quantification of PD-L1+ cells. As cancer segmentation is a challenging step for TPS generation, we trained DeepLab V3 in the Visiopharm software package to outline cancer regions in PD-L1 and NC images and evaluated the model performance by mean intersection over union (mIoU) against manual outlines. Only 14 cases were required to accomplish a mIoU of 0.82 for cancer segmentation in hematoxylin-stained NC cases. For PD-L1-stained slides, a model trained on PD-L1 tiles augmented by registered NC tiles achieved a mIoU of 0.79. In segmented cancer regions from whole slide images, the digital TPS achieved an accuracy of 75% against the manual TPS scores from the pathology report. Major reasons for algorithmic inaccuracies include the inclusion of immune cells in cancer outlines and poor nuclear segmentation of cancer cells. Our transparent and stepwise approach and performance metrics can be applied to any IHC assay to provide pathologists with important insights on when to apply and how to evaluate commercial automated IHC scoring systems.

BackgroundS100A8 is a melanoma biomarker expressed in the melanoma-associated epidermal keratinocytes, but its diagnostic utility has not been compared with other biomarkers, including PRAME.ObjectivesTo compare the utility of S100A8 and PRAME immunohistochemistry (IHC) in the differential diagnosis of melanoma and naevi in a case-control study.MethodsA previously described cohort of 209 melanomas (case samples) and naevi (control samples) dual-immunostained for S100A8 and PRAME were included. For S100A8, previously reported scores indicating the proportion of tumour-associated epidermis stained (0 = indeterminate; 1 = 0-4%; 2 = 5-25%; 3 = 26-50%; 4 = 51-75%; 5 = > 75%) were utilized. PRAME IHC was reviewed by at least two reviewers and a consensus score assigned, with score indicating the proportion of tumour stained (0 = indeterminate; 1 = 0%; 2 = 1-50%; 3 = > 50%). A positive test was defined as > 50% staining.ResultsThe area under the receiver operating characteristic curves for S100A8 (0.833) and PRAME (0.874) were not significantly different from each other (P = 0.22). The diagnostic sensitivity and specificity were 42.4% [95% confidence interval (CI) 32.6-52.8%] and 98.2% (95% CI 93.6-99.8%) for S100A8, and 79.8% (95% CI 70.5-87.2%) and 87.3% (95% CI 79.6-92.9%) for PRAME, respectively. A combined test requiring both S100A8 and PRAME IHC positivity had a sensitivity of 39.4% (95% CI 29.7-49.7%) and specificity of 99.1% (95% CI 95.0-100.0%).ConclusionsS100A8 and PRAME have utility in the diagnostic workup of melanoma, with S100A8 being more specific and PRAME being more sensitive when using this threshold. Our findings suggest that these two immunohistochemical markers may favourably complement one another to improve the detection of melanoma.

Salivary glands have essential roles in maintaining oral health, mastication, taste and speech, by secreting saliva. Salivary glands are composed of several types of cells, and each cell type is predicted to be involved in the carcinogenesis of different types of cancers including adenoid cystic carcinoma (ACC), acinic cell carcinoma (AciCC), salivary duct carcinoma (SDC), myoepithelial carcinoma (MECA) and other histology. In our study, we performed single nucleus RNA-seq on three human salivary gland samples to clarify the gene expression profile of each complex cellular component of the salivary glands and related these expression patterns to expression found in salivary gland cancers (SGC) to infer cell of origin. By single nucleus RNA-seq, salivary gland cells were stratified into four clusters: acinar cells, ductal cells 1, ductal cells 2 and myoepithelial cells/stromal cells. The localization of each cell group was verified by IHC of each cluster marker gene, and one group of ductal cells was found to represent intercalated ductal cells labeled with HES1. Furthermore, in comparison with SGC RNA-seq data, acinar cell markers were upregulated in AciCC, but downregulated in ACC and ductal cell markers were upregulated in SDC but downregulated in MECA, suggesting that markers of origin are highly expressed in some SGC. Cell type expressions in specific SGC histology are similar to those found in normal salivary gland populations, indicating a potential etiologic relationship.

Introduction: We aim to share the experience of a single center in the management of eight cases of renal primitive neuroectodermal tumor (PNET) which are uncommon, aggressive tumors. The objectives were to study the presentation of the disease, the treatment offered and its outcomes, and the comparison of the treatment with published literature. Methods: The single-center renal PNET data of all patients were retrospectively reviewed from 2011 to 2022. Renal PNET was seen in eight patients. Minimum follow-up period of 1 year was required. Results: Male-to-female ratio was 7:1. The mean age was 26.5 years. All were locally advanced tumors on presentation. One patient had an inferior vena cava thrombus, one patient had metastases on presentation, and two patients had tumor extending to paranephric space. The diagnosis was made by histopathology supported by immunohistochemistry showing CD99 positivity. All patients were treated with radical nephrectomy, followed by chemotherapy in all and radiotherapy in three patients. Two patients expired at 3½ and 6 years after surgery, the remaining six are alive at a median follow-up period of 34.5 months. Conclusion: Renal PNET is an uncommon renal tumor which is aggressive and requires multimodal therapy for prolonged survival., Author(s): Niramya J. Pathak (corresponding author) [1]; Abhishek G. Singh [1]; Pavan J. Surwase [1]; Sahil A. Agrawal [1]; Arvind P. Ganpule [1]; Ravindra B. Sabnis [1]; Mahesh R. Desai [...]

Purpose: To investigate bone regeneration among three different bone graft materials in a rat calvarum model. Materials and Methods: A total of 24 rats had two 5-mm defects placed per calvarial. Rats were divided into four groups: bovine xenograft (XG), demineralized bone matrix (DBM), mineralized bone graft (MBG), and collagen membrane control (CC). Within each group, samples were collected at two time points: 4 weeks (T4) and 8 weeks (T8). Bone regeneration was assessed by microcomputed tomography (micro-CT) imaging and was analyzed using MATLAB software. Additionally, the fixed samples were subsequently demineralized for immunohistochemistry and histomorphometry. Slides were mounted and stained with hematoxylin and eosin (H&E) stain as well as bone morphogenetic protein 2 (BMP-2) and runt-related transcription factor 2 (RUNX2) markers. The numbers of positive cells/area were calculated for each group and analyzed. Results: At 4 weeks, DBM showed low mineral density (7.7%) compared to the control (25.2%), but increased dramatically at 8 weeks (DBM, T8 = 27.6%; CC, T8 = 27.2%). Xenograft material showed an increase in mineral desnity between T4 and T8 (XG, T4 = 25.0%; XG, T8 = 32.3%). MBG remained consistent over the 8-week trial period (MBG, T4 = 30.4%; MBG, T8 = 30.4%). BMP-2 expression was present in cells adherent to all graft materials. RUNX2 expression was also observed in cells adherent to all graft materials, indicating that during the 4- to 8-week healing period, all materials supported osteogenesis. Conclusions: Compared to other materials, the DBM had high osteoinductive properties during the 4- to 8-week time period based on increased mineral content. All materials were associated with immunohistologic evidence of osteogenesis in the rat calvarial defect model. [ABSTRACT FROM AUTHOR]

Purpose: To analyze the process of early oral osseointegration of titanium (Ti) implants in diabetic 129/Sv mice through microCT and histologic and immunohistochemical analysis. Materials and Methods: A group of 30 male 129/Sv mice was equally subdivided into two groups: (1) nondiabetic (ND), in which mice did not undergo systemic alterations and received a standard diet, and (2) diabetic (D), in which mice were provided a high-fat diet from the age of 6 weeks until the conclusion of the study and received two intraperitoneal (IP) injections of streptozotocin (STZ) at a concentration of 100 mg/Kg each. Each mouse underwent extraction of a maxillary first molar, and customized Ti screws (0.50 mm diameter, 1.5 mm length) were placed in the residual alveolar sockets of the palatal roots. At 7 and 21 days after implant placement, the animals were euthanized for maxilla and pancreas collection. Maxillae containing Ti implants were analyzed with microCT, histology, and immunohistochemistry for cells that were positive for F4/80, CD146, runt-related transcription factor 2 (Runx2), and proliferating cell nuclear antigen (PCNA). Pancreata were histologically analyzed. Quantitative data were statistically analyzed with a significance level at 5% (P < .05). Results: ND mice presented successful healing and osseointegration, with a significantly higher fraction of bone volume compared to D mice, both at the alveolar sockets (53.39 ± 5.93 and 46.08 ± 3.18, respectively) and at the implant sites (68.88 ± 7.07 and 44.40 ± 6.98, respectively) 21 days after implant placement. Histologic evaluation revealed that the ND mice showed a significant decrease in inflammatory infiltrate and a significant increase in newly formed bone matrix at 21 days, whereas peri-implant sites in the D mice were predominantly encapsulated by fibrous tissue and chronic inflammatory infiltrate. Immunohistochemical characterization revealed higher Runx2 osteoblast differentiation and higher cell proliferation activity in the ND mice at 7 days, while higher amounts of macrophages were present in D mice at 7 and 21 days. Interestingly, no differences were found in CD146-positive cells when comparing ND and D mice. Conclusions: This study evaluated the effects of immediate dental implant placement in 129/Sv diabetic mice by using specific healing markers to identify changes in cellular events involved in early oral osseointegration. This approach may serve as tool to evaluate new materials and surface coatings to improve osseointegration in diabetic patients. [ABSTRACT FROM AUTHOR]

Breast cancer is one of the most common malignancies that can lead to cutaneous metastasis. Dermatopathologists often play an important role in the diagnosis of breast cancer metastasis to the skin. Rarely, dermatopathologists render a histopathologic diagnosis of primary breast cancer. We discuss a 51-year-old man with metastatic breast adenocarcinoma who presented after admission to the intensive care unit in the setting of altered mental status and critical anemia. Examination revealed a 14cmx12cm ulcerated, fungating tumor occupying the left breast. A four mm punch biopsy from the mass showed cords of atypical cells infiltrating the mid-to-deep dermis positive for CK7, GATA3, ER and PR. CK20, P40, p63, and TTF1 stains were negative. HER2/NEU immunoperoxidase stain was negative. CA15-3 was elevated at 75U/ml. Taken together, he was diagnosed with primary left breast ductal adenocarcinoma, grade two with subsequent visceral metastases to the bones, lymph nodes, and lungs. Although male breast cancer makes up less than 1% of all breast cancers, its incidence has been increasing worldwide. Recognition of the unique clinical and histologic findings of primary breast carcinoma is important to avoid delay in the diagnosis and initiation of appropriate treatment.

Considering the lack of consensus related to the impact of selective IL-6 receptor inhibition on bone remodeling and the scarcity of reports, especially on large bone defects, this study proposed to evaluate the biological impact of the selective inhibitor of interleukin-6 receptor (tocilizumab) in an experimental model of critical calvarial defect in rats. In this preclinical and in vivo study, 24 male Wistar rats were randomly divided into two groups (n=12/group): defect treated with collagen sponge (CG) and defect treated with collagen sponge associated with 2 mg/kg tocilizumab (TCZ). The defect in the parietal bone was created using an 8-mm diameter trephine drill. After 90 days, the animals were euthanized, and tissue samples (skull caps) were evaluated through micro-CT, histological, immunohistochemistry, cytokines, and RT-qPCR analyses. Tocilizumab reduced mononuclear inflammatory infiltration (P0.05). The bone cells (osteoblasts, osteoclasts, and osteocytes) in the defect area were similar in both groups (P>0.05). Tocilizumab reduced inflammatory cytokines, decreased osteogenic protein, and increased proteases in a critical bone defect in rats. Ninety days after the local application of tocilizumab in the cranial defect, we did not find a significant formation of bone tissue compared with a collagen sponge.

Novel brain clearing methods revolutionize imaging by increasing visualization throughout the brain at high resolution. However, combining the standard tool of immunostaining targets of interest with clearing methods has lagged behind. We integrate whole-mount immunostaining with PEGASOS tissue clearing, referred to as iPEGASOS (immunostaining-compatible PEGASOS), to address the challenge of signal quenching during clearing processes. iPEGASOS effectively enhances molecular-genetically targeted fluorescent signals that are otherwise compromised during conventional clearing procedures. Additionally, we demonstrate the utility of iPEGASOS for visualizing neurochemical markers or viral labels to augment visualization that transgenic mouse lines cannot provide. Our study encompasses three distinct applications, each showcasing the versatility and efficacy of this approach. We employ whole-mount immunostaining to enhance molecular signals in transgenic reporter mouse lines to visualize the whole-brain spatial distribution of specific cellular populations. We also significantly improve the visualization of neural circuit connections by enhancing signals from viral tracers injected into the brain. Last, we show immunostaining without genetic markers to selectively label beta-amyloid deposits in a mouse model of Alzheimers disease, facilitating the comprehensive whole-brain study of pathological features.

Endothelial cells of mammalian blood vessels have multiple levels of heterogeneity along the vascular tree and among different organs. Further heterogeneity results from blood flow turbulence and variations in shear stress. In the aorta, vascular endothelial protein tyrosine phosphatase (VE-PTP), which dephosphorylates tyrosine kinase receptor Tie2 in the plasma membrane, undergoes downstream polarization and endocytosis in endothelial cells exposed to laminar flow and high shear stress. VE-PTP sequestration promotes Tie2 phosphorylation at tyrosine992 and endothelial barrier tightening. The present study characterized the heterogeneity of VE-PTP polarization, Tie2-pY992 and total Tie2, and claudin-5 in anatomically defined regions of endothelial cells in the mouse descending thoracic aorta, where laminar flow is variable and IgG extravasation is patchy. We discovered that VE-PTP and Tie2-pY992 had mosaic patterns, unlike the uniform distribution of total Tie2. Claudin-5 at tight junctions also had a mosaic pattern, whereas VE-cadherin at adherens junctions bordered all endothelial cells. Importantly, the amounts of Tie2-pY992 and claudin-5 in aortic endothelial cells correlated with downstream polarization of VE-PTP. VE-PTP and Tie2-pY992 also had mosaic patterns in the vena cava, but claudin-5 was nearly absent and extravasated IgG was ubiquitous. Correlation of Tie2-pY992 and claudin-5 with VE-PTP polarization supports their collective interaction in the regulation of endothelial barrier function in the aorta, yet differences between the aorta and vena cava indicate additional flow-related determinants of permeability. Together, the results highlight new levels of endothelial cell functional mosaicism in the aorta and vena cava, where blood flow dynamics are well known to be heterogeneous.

Background: Nasopharyngeal carcinoma (NPC) is a highly invasive malignant tumor. Recurrence and distant metastasis represent the primary causes of treatment failure. This study aimed to identify biomarkers highly associated with NPC and investigate its roles in tumor progression. Methods: Transcriptome sequencing (RNA-seq) data of NPC and normal tissues were downloaded from the Gene Expression Omnibus (GEO) database. By analyzing the RNA-seq data, we found that G Protein Subunit Alpha 14 (GNA14) is closely associated with the diagnosis and prognosis of NPC. Immunohistochemistry (IHC) was used to detect the expression of GNA14 in tumor tissues of 165 NPC patients, and we analyzed the relationship between GNA14 expression and patient prognosis. The potential mechanisms by which GNA14 affects tumor prognosis were preliminarily analyzed using bioinformatics analysis. Results: Analysis of RNA-seq data and IHC showed that GNA14 expression was downregulated in NPC (p < 0.001, p < 0.01, respectively), and low expression of GNA14 was closely associated with poor prognosis. IHC analysis showed that patients with low GNA14 expression had significantly shorter progression-free survival (PFS) and distant metastasis-free survival (DMFS) than those with high GNA14 expression (p = 0.023, p = 0.008, respectively). Multivariate analysis indicated that the GNA14 expression was an independent risk factor for DMFS (p = 0.030). The DMFS nomogram included GNA14 expression, EBV DNA, and N stage as prognostic factors and the concordance index (C-index) of the nomogram was 0.73. Bioinformatics analysis indicated that NPC patients with low GNA14 expression might represent lower levels of immune cell infiltration and poorer drug sensitivity. Conclusion: Low GNA14 expression may be a risk factor for poor prognosis in NPC. [ABSTRACT FROM AUTHOR]

Background: Papillary thyroid carcinoma (PTC) is the most common type of thyroid carcinoma, representing the majority of thyroid cancer cases. Most patients with PTC have an excellent prognosis following treatment, yet approximately 10% face mortality within ten years, primarily due to lymph node metastasis (LNM) or local recurrence. The SIX1 gene, a member of the SIX gene superfamily, encodes a transcription factor integral to the development of certain tissues during embryogenesis. The impact of SIX1 in different subtypes of PTC has not been studied previously. Objective: The purpose of this study was to investigate the expression of SIX1 protein in PTC and to explore its relationship with clinical behavior in two subtypes of PTC: classic PTC (C-PTC) and follicular variant PTC (FV-PTC). Materials and methods: Using immunohistochemistry, the study analyzed 125 primary PTC cases, including 85 cases of C-PTC and 40 cases of FV-PTC. Results: The study found significant positive associations between high SIX1 expression and several adverse clinical features across the PTC samples. High SIX1 expression was linked with increased tumor size, multifocal tumors, LNM, high-grade tumor features, advanced tumor stage, lymphovascular invasion, perineural invasion, and extrathyroidal extension (ETE). Within the classic PTC subgroup, high SIX1 expression showed significant positive correlations with Tumor size (P = 0.04), Multifocality (P = 0.02) and High-grade features (P = 0.03). In the follicular variant subgroup, high SIX1 expression was significantly associated with Lymph node metastasis (LNM) (P = 0.001), Lymphovascular invasion (P = 0.03), ETE (P = 0.003) and tumor stage (P = 0.007). Conclusions: The findings of this study indicate that SIX1 expression is a marker of poor prognosis in PTC, suggesting that its high expression is linked with more aggressive tumor characteristics and advanced disease stages. Importantly, the impact of SIX1 expression varies between C-PTC and FV-PTC, predicting distinct prognostic factors in each subtype. This suggests that SIX1 could be utilized not only as a prognostic biomarker but also in developing subtype-specific therapeutic strategies for PTC patients. [ABSTRACT FROM AUTHOR]

ABSTRACT Objectives Methods Results Conclusions To define tumor immunoarchitectural patterns (IPs) and characterize the immune profile in salivary gland mucoepidermoid carcinoma (MEC) toward assessing MEC prognostic significance and implications for immunotherapy.This study analyzed 41 MEC cases, evaluating the tumor IPs and tumor‐infiltrating lymphocyte (TIL) parameters by using whole‐slide imaging and AI‐assisted assessment. Immunohistochemistry of CD3 and CD8 markers was performed to assess key lymphocyte subpopulations.Immune‐rich (I‐rich) tumors were characterized by high TIL density and were associated with aggressive tumor behavior, particularly in histological high‐grade MEC, with a significant increase in TIL density observed in the inner invasive margin compartment (p < 0.05). I‐rich tumor cases were linked to poorer disease‐free survival (DFS) (Breslow = 4.686, p = 0.03). Aggressive MEC cases displayed a highly heterogeneous TIL profile, however, no TIL patterns showed a significant impact on DFS (p > 0.05).Despite the high immunogenicity suggested by the abundance of T lymphocytes, the protective role of CD8+ T lymphocytes was not prominent in MEC. I‐rich tumors appeared more aggressive and unfavorable MEC cases exhibited high heterogeneity in their TIL profiles. Interrogating the role of TILs in the inner invasive tumor margin may offer a new mechanistic understanding for future immunotherapy discovery. [ABSTRACT FROM AUTHOR]

Background: Oral leiomyomatous hamartoma (OLH) corresponds to an abnormal, benign and disorganized overgrowth of mature smooth muscle tissue, which can develop in any site where this tissue is found in healthy conditions. The present systematic review aimed to analyze the clinicopathological characteristics of OLH. Materials and methods: The protocol of this study was constructed following the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) and was registered in the Open Science Framework (OSF): OSF.IO/BMPUX. Five electronic databases were used to identify studies for this systematic review: PubMed, Web of Science, Dentistry & Oral Science Source, Scopus and ScienceDirect, from January 15th, 1945 to January 10th, 2024. The Joanna Briggs Institute (JBI) tool was used to assess the risk of bias and the quality of the included reports and case series. Results: A comprehensive search yielded 5,562 articles, of which 55 met the inclusion criteria. The total number of subjects studied in the included investigations was sixty-six. The subjects' ages varied from newborns to 61 years, with a mean age ± standard deviation of 71.23 ± 123.01 months. 50.7% were males and 49.3% were females. Most lesions presented normochromic color (24.24%), pedunculated base (31.81%), firm consistency (22.72%), with an average size of 1.20 cm, present on the dorsum of the tongue (31.81%) or anterior part of the alveolar border of the maxilla (30.30%). Treatment was carried out by surgical excision (78.46%) and half of the studies report that there were no recurrences. Conclusions: The cases described in the medical-dental literature provide valuable information to date on the clinicopathologic and immunohistochemical profile of OLH. Although it is a rare lesion, it should be considered as part of the differential diagnosis in newborns, infants, children and young adults with lingual and/or maxillary masses present in the midline. [ABSTRACT FROM AUTHOR]

Background: p53 is a tumor suppressor gene. p53 expression in epithelial ovarian tumors (EOTs) is correlated with their biological behavior and predicts patient overall survival. However, there is a dearth of knowledge regarding p53 expression in these tumors among women from southwest Nigeria. Our study aimed to determine the patterns of p53 expression in various types of epithelial ovarian tumours. Methods: We conducted a retrospective study of epithelial ovarian tumours. We retrieved formalin-fixed, paraffin-embedded (FFPE) tissue blocks of previously diagnosed epithelial tumors from the departmental archive. We performed immunohistochemical analysis using p53 antibodies. We scored the expression and staining intensity of p53 as follows: negative (0), focal/weakly positive (1 +), and diffuse/strongly positive (2 +) on the basis of the recommended Cytomation scoring system. Results: The spectrum of p53 expression in the 51 histologically diagnosed cases revealed that 29 cases had no expression, consisting of 21 benign EOTs, two borderline EOTs, and six malignant EOTs. Nine cases exhibited wild-type expression, including six serous carcinomas, two mucinous carcinomas, and one signet ring cell carcinoma. p53 overexpression was observed in 13 patients overall, with 12 having serous carcinomas and one having endometrioid carcinoma. Among the 21 serous carcinoma patients, 28.6% (6 patients) presented with wild-type p53 expression, 57.1% (12 patients) presented with p53 overexpression, and 14.3% (three patients) presented negative p53 expression. There was a significant association between p53 expression and the histological grade of serous carcinoma. Conclusion: Most epithelial ovarian carcinomas in our hospital are high grade, with many serous carcinomas showing either p53 overexpression or loss of expression. This may contribute to the poor patient survival rate. [ABSTRACT FROM AUTHOR]

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Colorectal cancer (CRC) ranks as the third most prevalent tumor and the second leading cause of mortality. Early and accurate diagnosis holds significant importance in enhancing patient treatment and prognosis. Machine learning technology and bioinformatics have provided novel approaches for cancer diagnosis. This study aims to develop a CRC diagnostic model based on immunohistochemical staining image features using machine learning methods. Initially, CRC disease-specific genes were identified through bioinformatics analysis, SVM-RFE and Random Forest algorithm utilizing RNA-seq data from both GEO and TCGA databases. Subsequently, verification of these genes was performed using proteomics data from CPTAC and HPA database, resulting in identification of target proteins (AKR1B10, CA2, DHRS9, and ZG16) for further investigation. SVM and CNN were then employed to analyze and integrate the characteristics of immunohistochemical images to construct a reliable CRC diagnostic model. During the training and validation process of this model, cross-validation along with external validation methods were implemented to ensure accuracy and reliability. The results demonstrate that the established diagnostic model exhibits excellent performance in distinguishing between CRC and normal controls (accuracy rate: 0.999), thereby presenting potential prospects for clinical application. These findings are expected to provide innovative perspectives as well as methodologies for personalized diagnosis of CRC while offering more precise references for promising treatment. [ABSTRACT FROM AUTHOR]

Acute cellular rejection (ACR) frequently occurs following lung transplantation (LuTx) and represents a risk factor for the development of chronic lung allograft dysfunction (CLAD) as well as long-term survival. The histopathological diagnosis of ACR carries a burden of interobserver variability. The widespread utilization and cost-effectiveness of immunohistochemistry (IHC) was proven beneficial in diagnosing rejection in human kidney transplantations and LuTx rat models. However, its potential for ACR detection in patients remains unexplored. We analyzed surface markers (CD3, CD4, CD8, CD20, CD68, CD47, PD-1, PD-L1, and CD31/PECAM-1) on lung tissue cryobiopsy samples collected within 6 months post-LuTx from 60 LuTx recipients, 48 of whom were diagnosed with ACR. Additionally, serum samples from 51 patients were analyzed using a multiplex bead-based Luminex assay. The cytokines and markers included PD-L1, IL2, TNFα, IFNγ, and Granzyme B. We observed a significant increase in PD-L1 tissue expression within the rejection group, suggesting a concerted effort to suppress immune responses, especially those mediated by T-cells. Furthermore, we noted significant differences in PECAM-1 levels between ACR/non-ACR. Additionally, peripheral blood C-reactive-protein levels tended to be higher in the ACR group, while Luminex serum analyses did not reveal any significant differences between groups. In conclusion, our findings suggest the potential value of PECAM-1 and PD-L1 markers in diagnosing ACR. [ABSTRACT FROM AUTHOR]

Introduction: Gastric cancer is the fifth most common cancer worldwide and the fourth most common cause of cancer-related death. Two molecular subtyping classifications were recently introduced: The Cancer Genome Atlas (TCGA) and the Asian Cancer Research Group (ACRG) classifications. Methods: We classified a cohort of 283 gastric cancer patients undergoing surgery at Helsinki University Hospital between 2000 and 2009. We constructed a tumour tissue microarray immunostained for the following markers: microsatellite instability (MSI) markers MSH2, MSH6, MLH1, and PMS2; p53; E-cadherin; and EBERISH. Results: In the univariate survival analysis for disease-specific survival, the Epstein–Barr virus (EBV) -positive subtype exhibited the worst prognosis with a hazard ratio (HR) of 2.49 (95% confidence interval [CI] 1.19–5.25, p = 0.016) compared with the most benign subtype, chromosomal instability (CIN). Using TCGA's classification, the genetically stable (GS) and MSI subtypes exhibited a worse survival compared with CIN (HR 1.73 [95% CI 1.15–2.60], p = 0.009 and HR 1.74 [95% CI 1.06–2.84], p = 0.027, respectively). Using the ACRG classification, the p53 aberrant subtype exhibited the best prognosis, whereas wild-type p53, MSI, and the epithelial–mesenchymal transition (EMT) subtypes exhibited poorer prognoses (EMT: HR 1.90 [95% CI 1.30–2.77], p < 0.001) when compared with aberrant p53. Conclusions: Immunohistochemical analysis can identify prognostically different molecular subtypes of gastric cancer. The method is inexpensive and fast, yet reveals significant information for clinical decision-making. However, our study did not find that either molecular subtyping performed better than the other classification. Thus, further development of the most optimal grouping of different molecular subtypes is still needed. [ABSTRACT FROM AUTHOR]

Background: The repair of articular cartilage defects is always a significant clinical challenge in joint treatment. Therefore, the aim of this study was to investigate that the ColII-HA-CS-HAP scaffolds with BMSCs could repair cartilage defects of knee. Methods: Bone marrow mesenchymal stem cells (BMSCs) were extracted from rabbits, identified using immunofluorescence staining, and successfully induced into chondrocytes. Type II collagen (ColII) was isolated from bovine cartilage and constructed into scaffolds with hyaluronic acid, chondroitin sulfate, and hydroxyapatite. Then BMSCs were seeded on the ColII-HA-CS-HAP scaffold to detect biocompatibility. Results: The results of DAPI fluorescence staining showed that the number of BMSCs on the ColII-HA-CS-HAP scaffolds increased rapidly after culturing for 12 d. The rabbit knee cartilage defect model with a diameter of approximately 3 mm and a thickness of approximately 4 mm was selected to evaluate the regenerative potential of the scaffolds using histological and immunohistochemical analyses. At 6 months, the regenerated cartilage in the ColII-HA-CS-HAP scaffolds with BMSCs was more similar to that of native cartilage than the ColII-HA-CS-HAP scaffold group. Conclusions: Our study proved that the ColII-HA-CS-HAP scaffolds with differentiated BMSCs can produce an excellent healing response and repair cartilage defects successfully in a rabbit model. [ABSTRACT FROM AUTHOR]

Objective: To explore the therapeutic effects of phellodendrine on non-compression lumbar disc herniation (NCLDH). Methods: The Sprague Dawley rat model of NCLDH was established via autologous caudal nucleus pulposus transplantation. Behavioral observations and neurological function scoring were conducted in Sprague Dawley rats, and the serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured via enzyme-linked immunosorbent assay (ELISA). Real-time quantitative polymerase chain reaction (RT‒qPCR) was used to detect the expression of nuclear factor kappa-B (NF-κB) p65 mRNA in L5 nerve roots and surrounding tissues. Western blotting was used to assess the protein expression of NF-κB p65 and TNF-α. Immunofluorescence and immunohistochemical analyses were performed to investigate the distribution and expression of the NF-κB p65 protein in the L5 nerve and its surrounding tissues. Results: In this animal study, phellodendrine was found to downregulate the expression of p65 mRNA, decrease the release of inflammatory factors, and alleviate motor dysfunction caused by lumbar disc herniation(LDH). Therefore, the phellodendrine technique has potential value for the treatment of NCLDH. Conclusion: In this animal experiment, phellodendrine was found to significantly reduce the expression level of p65 mRNA, decrease the release of inflammatory cytokines, and alleviate lumbar disc pain. Clinical trial number: Not applicable. [ABSTRACT FROM AUTHOR]

Transient abnormal myelopoiesis (TAM) in neonates with Down syndrome (DS) exhibits hematological features that are indistinguishable from those of acute megakaryoblastic leukemia. However, TAM typically resolves spontaneously within several months postnatally. Some patients with TAM, however, develop severe clinical manifestations, which can lead to an unfavorable prognosis. TAM originates in utero through the acquisition of somatic GATA1 mutations, resulting in the loss of the full‐length GATA1 protein and excessive production of the N‐terminal truncated short isoform of the GATA1 protein (GATA1s). Herein, we report the pathological findings from an autopsy of a female stillbirth with TAM and DS, including an examination of her placental tissues. Immunohistochemical analysis revealed the expression of GATA1s, but not full‐length GATA1, in CD42b‐positive atypical immature megakaryocytes and blasts in the placental blood vessels. This confirms the diagnosis of TAM and suggests the utility of placental tissue for histological diagnosis. Additional unique findings from the autopsy specimens are discussed. [ABSTRACT FROM AUTHOR]

Sweat dermatitis is an irritant inflammatory dermatosis commonly encountered in tropics during summers. Profuse sweating triggered by high heat index, friction, clothing, outdoor activities, military training, overcrowding, prolonged working in ill-ventilated places, etc., are responsible or this dermatitis. Aim was to study the clinical, dermoscopic, vapometric, histopathological, and immunohistochemistry (IHC) findings in sweat dermatitis. Twenty-five patients of sweat dermatitis attending out-patient department (OPD) of tertiary care hospital, were recruited over a period of three months (June to August 2022). A detailed history was followed by dermatological examination, dermoscopy, vapometry, histopathology, and IHC studies. Twenty-five patients were included in the study. There was male preponderance with male: female ratio of 2.1:1. The mean age of patients was 30.36 years. Dermatological examination in all patients revealed brownish "parchment"-like, shiny "crinkled cellophane paper"-like patches over the entire upper back and shoulder region with areas of exfoliation. Dermoscopy revealed increased skin markings, deep brown pigmentary changes with superimposed whitish scales with islands of eroded skin, and prominent eccrine duct openings. Vapometric analysis revealed increased transepidermal water loss (TEWL) in the affected area in 88% of patients. Histopathological examination showed features of spongiotic dermatitis. IHC with CK7 (cytoplasmic) highlighted the normal eccrine coils. Sweat dermatitis is common seasonal dermatoses in tropics, but it remains undiagnosed due to lack of awareness. This study aims to sensitize medical doctors and dermatologists about sweat dermatitis and prevention strategies that can be helpful in mitigating it. The study also discusses the role of investigative modalities, such as dermoscopy, vapometry, histopathology, and IHC, in sweat dermatitis. [ABSTRACT FROM AUTHOR]

Objective: Claudin18.2 has been established as a putative therapeutic target in human solid malignancies. The aim of this study is to determine claudin18.2 expression as a clinicopathological and prognostic factor in human solid tumors through a systematic review and meta-analysis. Articles were systematically reviewed for studies that included the correlation between claudin18.2 expression and clinicopathological features and prognosis in solid tumors. Meta-analysis was conducted to estimate either odds ratio and 95% confidence intervals (CIs) of clinicopathological factors or hazard ratio and 95% CIs of survival outcomes for claudin18.2 expression in all available solid tumors. Results: 21 studies including 5,331 patients were identified. Overall proportion of claudin18.2 positivity was 29.7%. Analyses of clinicopathological features demonstrated that claudin18.2 positivity correlated with male predominance, lower T stage, more frequent MUC5AC positivity when all primary tumors included. In subgroup analysis, gastric cancer showed significant correlation between high claudin18.2 expression and frequent EBV infection, male predominance and lower T stage. In lung cancer, claudin18.2 expression was associated with favorable overall survival. However, analyses of survival outcomes in all solid tumors showed that claudin18.2 expression was not associated with overall survival and pooled disease-free survival, tumor-specific survival, progression-free survival and relapse-free survival. Conclusions: Our study emphasizes evaluation of claudin18.2 expression as a potential prognostic factor in lung adenocarcinoma and further exploration in other solid tumors as well. Systematic review registration: https://www.crd.york.ac.uk/prospero/ , identifier CRD42023468651. [ABSTRACT FROM AUTHOR]

Primary squamous cell carcinoma (SCC) can originate in different parts of the body, including the head, neck, lung, bronchus, cervix uteri, esophagus, and cardia, and subsequently metastasize to the stomach. Primary gastric squamous cell carcinoma (GSCC) is a rare disease. To better understand GSCC, we present the case of a 72-year-old woman with a primary GSCC. A chest and abdominal CT scan highlighted a 36×26 mm mass with a 41 mm longitudinal diameter, which included the origin of the celiac tripod. The disease appeared to originate exophytically from the gastric wall. An ultrasound-endoscopy showed a hypoechoic formation with not well-defined margins measuring 40×30 mm involving the origin of the celiac tripod, about 10 mm from the gastric wall. An endoscopic fine-needle aspiration showed a poorly differentiated carcinoma. A PET/CT scan showed a hyperaccumulation of the known expansive formation at the celiac tripod (SUV 11.9) without specific cleavage planes from the stomach. A gastroscopy showed a regular esophagus and an absence of gastric protruding lesions. In the subcardial area, on the posterior wall, there was a slightly raised sub-centimetric area covered by bleeding mucosa where the biopsy had been performed. The pathological report showed chronic gastritis. An eco-endoscopy confirmed a hypoechoic neoformation measuring 30×40 mm that appeared to originate from the muscular layer of the gastric wall. The biopsy report was positive for broad-spectrum cytokeratins (AE1/AE3), CK5/6/7, p40, p63 and negative for CK20, PAS, TTF-1, anti-smooth muscle actin, CD45 (LCA), ERG, and S100. The clinical picture suggested poorly differentiated carcinoma with squamous differentiation. We analyzed the main classifications of GSCC cases and compared their characteristics. It is clear that to have an appropriate definition of GSCC, well-defined diagnostic criteria are needed. Currently, there is no consensus. For practical purposes, it would be better to include a panel of CK and p40 to distinguish GSCC from adenocarcinoma. A GSCC outside the mucosa is not rare and could be a true entity. [ABSTRACT FROM AUTHOR]

Gastrointestinal stromal tumors (GISTs) arise from the interstitial cells of Cajal, predominantly affecting the stomach and small intestine, with rare occurrences in the duodenum, rectum, and extraintestinal sites. Histologically, GISTs can present as spindle cells, epithelioid cells, or mixed morphologies, with immunohistochemical staining revealing expression of KIT (CD117) and discovered on GIST 1 (DOG1). Approximately 80% of GISTs harbor activating mutations in KIT or platelet derived growth factor receptor α (PDGFRA), which influence their clinical behavior and treatment response. SDH-deficient GISTs, associated with syndromes such as Carney triad and Carney–Stratakis syndrome, represent a distinct subgroup with unique characteristics and management challenges. The standard treatment includes surgery and imatinib for metastatic cases; however, resistance to tyrosine kinase inhibitors remains a significant hurdle, especially in pediatric and wildtype GISTs. This highlights the need for advanced therapeutic strategies and emphasizes the importance of molecular profiling in guiding treatment decisions and improving outcomes for GIST patients. [ABSTRACT FROM AUTHOR]

Endocrine mucin-producing sweat gland carcinoma is a rare neoplasm of the skin appendages. The tumor typically exhibits slow growth and rarely metastasizes to distant sites. Herein, we report a case of a 77-year-old male who presented with a skin lesion on the right anterior chest wall 23 years ago. Fifteen years later, surgical excision was performed, and the pathological diagnosis was endocrine mucin-producing sweat gland carcinomas. However, the histopathological examination revealed a coexistence of endocrine mucin-producing sweat gland carcinomas and mucinous carcinoma components. Over the past 2 years, the patient developed lymph node metastasis in the right axilla, local recurrence on the right chest wall, and distant skin metastasis. The histopathological type of the lymph node metastasis was consistent with the primary tumor, while the recurrent and skin metastatic lesions exhibited mucinous carcinoma. To our knowledge, this is the first reported case of endocrine mucin-producing sweat gland carcinoma with distant skin metastasis, characterized by two distinct carcinoma components in the histopathological morphology. [ABSTRACT FROM AUTHOR]

Background: Although activation of human epidermal growth factor receptor 3 (HER3) is linked to resistance to targeted therapies in several cancer types, the HER3 expression profile during pancreatic cancer treatment remains unknown. Aims: We evaluated the HER3 expression status after chemotherapy for pancreatic cancer and its association with clinicopathological features and clinical outcomes. Materials & Methods: We included patients with pancreatic cancer who underwent chemotherapy and whose post‐treatment archival tissue specimens were collected. HER3 expression was retrospectively assessed by immunohistochemistry scoring (0, 1+, 2+, and 3+) of the membranous staining intensity. Results: HER3 expression after chemotherapy was evaluated in 41 patients, with matched‐pair analysis in five patients before and after chemotherapy. HER3 expression was observed in most of the patients after chemotherapy, demonstrating IHC scores of ≥ 1+ and ≥ 2+ in 40 (98%) and 26 (63%) of 41 patients, respectively. Of the 38 patients with adenocarcinoma, the median overall survival in the HER3 (2+/3+) and HER3 (0/1+) groups was 21.0 and 17.1 months, respectively. The comparison of HER3 expression before and after chemotherapy performed in five cases revealed that scores changed from 2+/3+ to 0/1+ in one case, 0/1+ to 2+/3+ in another case, and remained at 2+/3+ in three cases. Cancer genome profiling tests in eight cases found no HER3 amplification or mutation, and seven of these cases had adenocarcinomas with KRAS and TP53 mutations. Conclusion: A high prevalence of HER3 expression was observed in pancreatic cancer patients after chemotherapy. Our findings indicate that HER3 is a potential therapeutic target for pancreatic cancer, deserving further clinical investigation. [ABSTRACT FROM AUTHOR]

Although diagnostic and therapeutic advances have been made in the treatment of breast cancer, the challenge of effectively controlling tumor progression persists. The objective of this study was to determine the relationship between IDO1 and TDO2 expression in breast cancer by immunohistochemistry and real-time polymerase chain reaction, hormone receptor status, Ki67 proliferation index, molecular classification, metastasis, and to investigate whether IDO1 and TDO2 expression can be used in combination with targeted therapy or as a marker to increase treatment efficacy in selected cases. The study included 74 cases of breast cancer and 14 cases of normal breast tissue as controls. All cases were analyzed for IDO1 and TDO2 by both immunohistochemistry and real-time polymerase chain reaction. The immunohistochemical analysis revealed that IDO1 immunoreactivity was significantly higher in tumor tissue compared to normal breast tissue. A statistically significant correlation was observed between IDO1 immunoreactivity and histologic subtypes. Furthermore, IDO1 gene expression was correlated with IDO1 immunoreactivity. TDO2 immune reactivity did not differ between tumor and non-tumor tissues and no correlation was found between histological subtypes. There was no correlation between TDO2 immunoreactivity and gene expression. The significant increase in IDO1 levels in tumor tissue and high positivity in age, HER2 positive and triple negative cases compared to other cases suggest that IDO1 inhibitors may be suitable for the target patient group in treatment selection. [ABSTRACT FROM AUTHOR]

The American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) recommend cold ischemia time (cIT) be <60 min, and formalin fixation time (FFT) 6–72 h, to optimize immunohistochemistry (IHC) based on breast cancer data. We assessed whether cIT and FFT impact IHC in endometrial cancer (EC), and determined which factors affect cIT and FFT. Surgical EC cases from 2019 to 2023 were reviewed. cIT was calculated by subtracting time of tissue devascularization intra-operatively from time the specimen was placed in formalin. Demographics, clinicopathologic and peri-operative factors, and IHC for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and mismatch repair (MMR) proteins were compared between patients with cIT <60 min versus ≥60 min (prolonged), and compliant FFT (6–72 h) versus non-compliant FFT (<6 or > 72 h). Categorical variables were compared using χ2 tests. 941 patients were included in the analysis. Median cIT was 33 min. Prolonged cIT occurred in 95 (10 %) cases. African American/Black race (p < 0.001), advanced stage (p < 0.001), mini-laparotomy (p < 0.001), performance of surgical procedures beyond standard EC staging (p < 0.001), longer surgical length (p < 0.001), and increased uterine weight (p < 0.001) were independently associated with prolonged cIT. There were no significant differences in ER, PR, HER2, or MMR protein expression based on cIT or FFT. Prolonged cIT was not associated with differences in biomarker expression via IHC at time of surgical staging for EC. Despite variability in cIT, which is largely due to non-modifiable factors, tumor molecular features remain consistent and can reliably be utilized for prognostic and therapeutic decision-making. • Cold ischemia time (cIT) is prolonged (≥60 min) in approximately 10 % of endometrial cancer cases. • Prolonged cIT is associated with Black race, advanced stage, mini-laparotomy, surgical length/extent, and uterine weight. • cIT is not associated with differences in ER, PR, HER2, MMR biomarker expression in endometrial cancer. [ABSTRACT FROM AUTHOR]

Genomic profiling-based model (GP-M) is the gold-standard for endometrial cancer (EC) molecular classification, but several issues related to the availability of genomic sequencing in low-income settings remain and health disparities in the management are increasing. This study aims to investigate the non-inferiority of the immunohistochemistry-alone model in classifying ECs compared to the standard genomic profiling-based model in terms of oncologic outcomes. All preoperative uterine-confined ECs undergoing surgical staging were retrospectively included. Patients classified by IHC-M were stratified into: MMR-proficient (MMRp), p53 wild type (p53wt) and estrogen receptor (ER) positive, 2) MMRp, p53wt and ER-negative, 3) MMRd, and 4) p53abn. A case-control comparison was performed between the IHC-M and GP-M cohorts. Then, a propensity-matched analysis was performed: ECs classified by IHC-M were matched in a 3:1 ratio with patients classified by GP-M. 1587 patients with EC were included. The Kaplan-Meier survival curves for disease-free survival and overall survival demonstrated that the two models performed similarly in risk-stratifying the study population (p < 0.0001). Moreover, the AUC-ROC showed overlapping results: 0.77 (0.66–0.87) for IHC-M and 0.72 (0.63–0.81) for GP-M, indicating that both models were able to successfully identify patients at high-risk and low-risk of disease recurrence/progression. The IHC-M showed overlapping classification performance compared to the GP-M in terms of oncologic outcomes. This study may lay the basis to further investigate the real-life clinical impact of POLE sequencing in molecular classification and the potential stand-alone prognostic role of ER status for further allocation of EC patients into risk classes. • The ProMisE model for endometrial cancer molecular classification is the standard. • However, genome sequencing has limited availability and cost-effectiveness. • Immunohistochemistry-alone model and ProMisE model predicted prognosis similarly. • Both models were able to successfully identify patients at high-risk of recurrence. • Estrogen receptor status should be integrated in the molecular classification model. [ABSTRACT FROM AUTHOR]

Background The cutoff of <1% positive cells to define estrogen receptor (ER) negativity by immunohistochemistry (IHC) in breast cancer (BC) is debated. We explored the tumor immune microenvironment and gene-expression profile of patients with early-stage HER2-negative ER-low (ER 1%-9%) BC, comparing them to ER-negative (ER <1%) and ER-intermediate (ER 10%-50%) tumors. Methods Among 921 patients with early-stage I-III, ER ≤50%, HER2-negative BCs, tumors were classified as ER-negative (n  =   712), ER-low (n  =   128), or ER-intermediate (n  =   81). Tumor-infiltrating lymphocytes (TILs) were evaluated. CD8+, FOXP3+ cells, and PD-L1 status were assessed by IHC and quantified by digital pathology. We analyzed 776 BC-related genes in 116 samples. All tests were 2-sided at a <.05 significance level. Results ER-low and ER-negative tumors exhibited similar median TILs, statistically significantly higher than ER-intermediate tumors. CD8/FOXP3 ratio and PD-L1 positivity rates were comparable between ER-low and ER-negative groups. These groups showed similar enrichment in basal-like intrinsic subtypes and comparable expression of immune-related genes. ER-low and ER-intermediate tumors showed significant transcriptomic differences. High TILs (≥30%) were associated with improved relapse-free survival (RFS) in ER-low (5-year RFS 78.6% vs 66.2%, log-rank P  =  .033, hazard ratio [HR] 0.37 [95% CI = 0.15 to 0.96]) and ER-negative patients (5-year RFS 85.2% vs 69.8%, log-rank P  <  .001, HR 0.41 [95% CI = 0.27 to 0.60]). Conclusions ER-low and ER-negative tumors are similar biological and molecular entities, supporting their comparable clinical outcomes and treatment responses, including to immunotherapy. Our findings contribute to the growing evidence calling for a reevaluation of ER-positive BC classification and management, aligning ER-low and ER-negative tumors more closely. [ABSTRACT FROM AUTHOR]

ABSTRACT: Gastrointestinal stromal tumors (GISTs) are rare mesenchymal tumors that arise from interstitial cells of Cajal. Due to vague presentation, location and confusing imaging studies, they tend to mimic gynaecological tumors. They usually diagnosed intra-operative and histopathology followed by tumor specific receptors such as KIT, CD34, CD 117 and DOG 1 are mainstay of diagnosis of GIST. Prognosis of GISTs depends on mitotic rate, tumor size and organ of origin. Resection of mass with tumor free margins is the target of treatment. L ymphadenectomy is not routine. Chemotherapy with tyrosine kinase inhibitors such as Imatinib, Dasatinib, Sorafenib and follow- up depend upon risk category. In this case series, there were four cases with vague symptoms misdiagnosed as gynaecological tumors are reviewed. Preoperatively tumors assumed to be of gynaecological origin were found to be case of GISTs intra-operatively and confirmed by presence of cajal's cells histopathology and mainly by DOG 1, CD117 and tyrosine kinase inhibitor receptors on immunohistochemistry. All belonged to high risk category of GISTs. Any abdomino-pelvic mass detected on ultrasonography and with unusual presentation presenting at primary health centre the possibility of non-gynecological tumors especially GISTs should be kept in mind and should be referred to higher centres for further investigation and proper management. [ABSTRACT FROM AUTHOR]

Many attempts have been made to simplify tissue fixation and processing prior to hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC) and genomic DNA extraction. The acetone-methyl benzoate-xylene (AMeX) method, is gaining popularity as an alternative to aqueous formaldehyde fixation followed by alcohol dehydration. We have assessed the efficacy of AMeX for the quality of extracted genomic DNA in addition to H&E and IHC in biopsy specimens of human oral mucosal lesions of size not more than 1 cm. H&E staining showed similar results when AMeX method was compared with conventional fixation and processing. Improved IHC staining quality was observed in the sections of AMeX-processed tissue, as were improved quality and quantity of genomic DNA. [ABSTRACT FROM AUTHOR]

The amygdala and raphe nuclei, which play crucial roles in mood regulation, are influenced by serotonergic neurotransmission. Alterations in this neurotransmission are associated with mood disorders. Therefore, using immunohistochemistry and quantitative methods this study was designed to evaluate potential alterations in the expression of serotoninergic markers in the amygdala and raphe nuclei of mice with oestrogen receptor β (ERβ) knock out which exhibit increased anxiety as evidenced by reduced locomotion and increased thigmotaxis. These alterations could either contribute to heightened anxiety or serve as a compensatory strategy for reducing it. The results show that in ERβ knock‐out mice, 5‐HT1B expression is significantly increased in the amygdala, while 5‐HTT expression is significantly decreased in both the amygdala and raphe nuclei. Furthermore, ERβ deficiency does not affect TPH2. In conclusion, serotonin signalling is altered in the amygdala and raphe nuclei of ERβ knock‐out females. It seems that an increase in 5‐HTT levels has been associated with reduced anxiety, whereas a decrease in 5‐HT1B receptors may encourage fear. However, further studies are required to determine the exact role of ERβ in anxiety‐related behaviour. [ABSTRACT FROM AUTHOR]

Surgical intervention for endometriosis is an important treatment modality, yet incomplete resection resulting from poor visibility of affected tissue and consequently recurrence of disease remains a prevalent challenge. Intra-operative visualization of endometriosis, enabling fluorescence-guided surgery (FGS), could help to optimize surgical treatment. A biomarker, upregulated in endometriosis compared to adjacent tissue, is required to use as a target for FGS. Immunohistochemistry was used to evaluate protein expression of a selection of previously identified potential biomarkers. Ten biomarkers were stained in a large cohort of 84 tissues, both deep and peritoneal endometriosis and tissue without endometriosis, all from patients with confirmed endometriosis. MMP11 and VCAN showed the largest upregulation in endometriosis compared to adjacent tissue and showed a membranous or extracellular staining pattern. MMP11 is a promising target for glandular and stromal visualization, VCAN for stromal visualization only. For both biomarkers, upregulation was high in both peritoneal and deep endometriosis and for patients with and without hormonal medication. Other stained biomarkers showed non-beneficial characteristics based on staining pattern or upregulation. Analysis of all endometriosis samples showed that combined glandular and stromal targeting is expected to result in optimal visualization of endometriosis. Further research is needed to determine whether targeting one biomarker is sufficient for this goal, or if dual targeting is necessary. Development of clinical tracers for VCAN and MMP11 is necessary. [ABSTRACT FROM AUTHOR]

Lynch syndrome (LS) is an autosomal dominant genetic disorder that results in an increased risk of ovarian and endometrial cancers. The aim of this paper was to explore the management of this risk through screening and prevention. Published materials and evidence were explored and summarized. This paper demonstrated that while there has been increased awareness and advances in the identification and diagnosis of patients with LS, recommendations for screening and prevention remain less evidence-based. In decisions of management of patients with LS, a shared decision-making model should be used considering individual patient goals. [ABSTRACT FROM AUTHOR]