Staining Pattern Research Articles

Perianal Paget Disease With an Enteric Phenotype and Associated In-Situ/Dysplastic Glandular Component: Report of a Potentially Novel Entity and Review of the Literature.

Extramammary Paget disease is an uncommon cutaneous malignancy that primarily affects areas rich in apocrine glands. Here, we aim to present an 84-year-old woman with a distinctive perianal neoplastic process comprised of conventional Paget disease with an intertwined in-situ glandular component. Rare foci of glands embedded in pools of mucin were also identified in the most recent excision, consistent with mucinous adenocarcinoma. Immunohistochemical staining demonstrated an enteric phenotype of the tumor cells, with expression of keratin 20, caudal type homeobox transcription factor 2 (CDX2), and special AT-rich sequence binding protein 2 (SATB2) (patchy, weak); keratin 7 and BerEP4 staining were also noted. Molecular analysis via next generation sequencing revealed pathogenic variants in ERBB2, TP53, and SF3B1. Given the unique histomorphology and immunohistochemical staining pattern, suspicion for a colorectal primary tumor was high, however; extensive workup including imaging, testing for tumor markers, and colonoscopic evaluation has not identified any other lesion thus far. Multiple biopsies and excisions in the area demonstrated recurrent disease over a 4-year span; with no evidence of deep invasion or metastasis. This tumor appears to be exceptional among reports in the literature given its extensive in-situ glandular component; and we are not aware of any documented SF3B1 mutation to date. Disease recurrence in this patient highlights the need for clinical vigilance. Defining the molecular profile in these lesions can also be useful, particularly when assessing potential treatment options for advanced disease.

Assessing pathogenicity of mismatch repair variants of uncertain significance by molecular tumor analysis

Functional analyses are the main method to classify mismatch repair (MMR) gene variants of uncertain significance (VUSs). However, the pathogenicity remains unclear for many variants because of conflicting results between clinical, molecular, and functional data. In this study, we evaluated whether whole exome sequencing (WES) could add another layer of evidence to elucidate the pathogenicity of MMR variants with conflicting interpretations. WES was performed on formalin-fixed paraffin-embedded tumor tissue of eight patients with a constitutional MMR VUS (seven families), including eight colorectal and two endometrial carcinomas and one ovarian carcinoma. Cell-free CIMRA assays were performed to assign Odds of Pathogenicity to these VUSs. In four families, seven tumors showed MMR deficiency-associated mutational signatures, supporting the pathogenicity of the VUS. Moreover, somatic (second) MMR hits identified in the WES data were found to explain MMR staining patterns when the MMR staining was discordant with the reported germline MMR gene variant. In conclusion, WES did not significantly reclassify VUS in these cases but clarified some phenotypic aspects such as age of onset and explanations in case of discordant MMR stainings.

Dual-ligand fluorescence microscopy enables chronological and spatial histological assignment of distinct amyloid-β deposits

Different types of deposits comprised of amyloid-β (Aβ) peptides are one of the pathological hallmarks of Alzheimer's disease (AD) and novel methods that enable identification of a diversity of Aβ deposits during the AD continuum are essential for understanding the role of these aggregates during the pathogenesis. Herein, different combinations of five fluorescent thiophene-based ligands were used for detection of Aβ deposits in brain tissue sections from transgenic mouse models with aggregated Aβ pathology, as well as brain tissue sections from patients affected by sporadic or dominantly inherited AD. When analyzing the sections with fluorescence microscopy, distinct ligand staining patterns related to the transgenic mouse model or to the age of the mice were observed. Likewise, specific staining patterns of different Aβ deposits were revealed for sporadic versus dominantly inherited AD, as well as for distinct brain regions in sporadic AD. Thus, by employing dual-staining protocols with multiple combinations of fluorescent ligands, a chronological and spatial histological designation of different Aβ deposits could be achieved. This study demonstrates the potential of our approach for resolving the role and presence of distinct Aβ aggregates during the AD continuum and pinpoints the necessity of using multiple ligands to obtain an accurate assignment of different Aβ deposits in the neuropathological evaluation of AD, as well as when evaluating therapeutic strategies targeting Aβ aggregates.

Examination of antinuclear antibody staining patterns and titers in patients with childhood-onset systemic lupus erythematosus.

Antinuclear antibodies (ANA) staining patterns can provide useful information in systemic lupus erythematosus (SLE). In our study, we examined the frequency of ANA staining patterns in disease-related features in childhood-onset SLE patients. ANA and its staining patterns were assessed in childhood-onset SLE patients. Two hundred twenty-three patients were included (F/M = 3/1). Their median age at diagnosis was 14.3 (11.9-16.1) years. The anti-cell (AC)-4/5 (fine or large speckled) pattern was the most common nuclear ANA pattern (75.8%), while the AC-19 (dense fine speckled) pattern was the most frequently detected cytoplasmic ANA pattern (13.1%). The AC-4/5 (fine or large speckled) patterns were notably seen in fever, acute and chronic cutaneous lupus, arthritis, serositis, hematologic involvement, renal involvement, neuropsychiatric involvement, gastrointestinal involvement, and cardiopulmonary involvement (all p < .001). Conversely, the AC-1 (homogeneous) pattern was significantly detected in oral/nasal ulcers and non-scarring alopecia (both p < .001). Regarding the laboratory features, the AC-4/5 (fine or large speckled) patterns exhibited a predominant seen in autoimmune hemolytic anemia, leukopenia, thrombocytopenia, elevated ESR and CRP, hypocomplementemia, direct Coombs, anti-Smith (Sm), anti-SSA and SS-B, anti-ribonucleoprotein (RNP), anti-histone, anti-ribosomal P, lupus anticoagulant, anti-cardiolipin immunoglobulin (Ig)M/IgG, and anti-β2-glycoprotein IgM/IgG positivities (all p < .001). In contrast, the AC-1 (homogeneous) pattern was detected in anti-double-stranded (ds) DNA and anti-histone positivity (both p < .001). Our study showed that AC-4/5 and AC-1 patterns of ANA are frequently detected in many clinical and serological features of childhood-onset SLE patients. However, further studies are needed in larger populations to verify these results.

The Utility of BSND Immunohistochemistry in the Differential Diagnosis of Oncocytic and Warthin-like Mucoepidermoid Carcinoma of Salivary Gland.

BSND is a chloride channel subunit that is expressed in the normal salivary gland. We aimed to validate the utility of BSND immunohistochemistry in the differential diagnosis of oncocytic salivary gland neoplasms. BSND immunohistochemistry was performed in a retrospective cohort of 93 salivary gland lesions, enriched with tumors with oncocytic features and histologic variants of mucoepidermoid carcinoma (MEC). All oncocytomas (n = 18) showed diffuse membranous BSND immunopositivity. Warthin tumors (n = 18) were also positive for BSND, but the staining pattern was patchy cytoplasmic and membranous in 10-25% of tumor cells. Using a threshold of 10% BSND-positive cells, all Warthin tumors were positive, while none of Warthin-like MECs or non-MEC salivary tumors were positive. Applying the same 10% positivity criterion, two oncocytic MECs were positive for BSND. The percentage of BSND staining in oncocytic MECs was up to 20%. In contrast, BSND was diffusely positive in oncocytomas with a median percentage of positivity of 95% (range: 40 - 100%). Therefore, a higher threshold of > 20% BSND-positive cells may be considered when differentiating between oncocytoma and oncocytic MEC. BSND immunohistochemistry is a potentially useful diagnostic marker for salivary gland neoplasms, especially oncocytic and Warthin-like MECs. A threshold of ≥ 10% positivity can differentiate Warthin tumors from Warthin-like MECs, whereas > 20% positivity can be effective for separating oncocytomas from oncocytic MECs.

Immunohistochemical examinations in malignant melanoma : Fundamentals and special aspects

Immunohistochemical examinations have been used for many years in dermatopathology and pathology and have become an integral part of tumor diagnostics. The aim is to identify and classify tumor cells that express distinct antigens. Malignant melanoma can be characterized by alarge number of well-described and standardized antibodies, so that immunohistochemical staining is used in the diagnosis of melanoma, the differential diagnosis of other tumors, the determination of tumor thickness, the diagnosis of tumor metastases, and also in the diagnosis of sentinel lymph nodes. Comprehensive knowledge of the expression profiles and specific staining patterns of the antibodies used is of great diagnostic relevance, with the aim of preventing misdiagnosis. The perfect antibody with high sensitivity and maximum specificity does not exist. Hence, immunohistochemistry does not replace the conventional assessment and interpretation of tumor samples. Only in symbiosis with clinical and conventional histological findings do immunohistochemical stains have adiagnostic value.

FAP-α is an effective tool to evaluate stroma invasion of lung adenocarcinoma

The main difficulty in the diagnosis of atypical in situ adenocarcinoma lies in the distinction between true and false stromal invasion. Moreover, how to identify local alveolar wall collapse in situ lung adenocarcinoma and how to identify whether the trapped adenoid structure around scar is an invasion component have become the key points for accurate diagnosis of lung adenocarcinoma. In the present study, we detected 40 cases of lung adenocarcinoma in situ and 40 cases of invasive adenocarcinoma by using immunohistochemical techniques. We found FAP-α had not immunreactivity in the stroma of adenocarcinoma in situ. However, it stained in the stroma of invasive areas in lung adenocarcinoma. FAP-α staining pattern could represent hyperplastic myofibroblast and demonstrated the true invasion of stroma. This study provides strong evidence that FAP-α is an effective tool to evaluate the presence or absence of stroma invasion of lung adenocarcinoma. Our findings will contribute to the accurate diagnosis of lung invasive adenocarcinoma.

Seromucinous hamartoma with unique clinical and histopathological features: a case report and review of the literature

BackgroundWe present this case to highlight the importance of considering seromucinous hamartoma in the differential diagnosis of nasal cavity lesions, particularly due to its rarity and potential for being mistaken for more aggressive pathologies. Seromucinous hamartoma, although benign, can exhibit clinical and histological features that overlap with those of malignant tumors, posing a diagnostic challenge. This case is especially noteworthy due to the unusual presentation of sebaceous differentiation within seromucinous hamartoma, a feature not previously documented in the literature. Recognizing such rare entities is crucial for ensuring appropriate patient management and avoiding unnecessary interventions.Case presentationWe report a unique case of a 41-year-old Caucasian man with seromucinous hamartoma, presenting with chronic left nasal obstruction and recurrent mild epistaxis. Nasal endoscopy revealed a well-defined, multilobulated lesion in the left nasal cavity. Computed tomography scans confirmed an anteromedial polypoid lesion, 31 mm × 15 mm. The lesion was endoscopically resected without complications, with no recurrence at 6-month follow-up. Pathological examination showed a filiform polypoid lesion with clusters of seromucinous glands, ducts, and tubules, and no invasive growth patterns. Immunohistochemical studies revealed distinct epithelial membrane antigen (EMA) and p63 staining patterns. Notably, mature sebaceous gland formation interspersed with seromucinous glands was observed, a novel finding in seromucinous hamartoma. Recognizing seromucinous hamartoma is crucial to avoid unnecessary treatments, and it should be included in differential diagnoses of nasal cavity lesions.ConclusionSeromucinous hamartoma is a rare and benign nasal cavity lesion that presents significant diagnostic challenges due to its potential to mimic more aggressive pathologies. This case highlights the importance of including seromucinous hamartoma in the differential diagnosis of sinonasal lesions, particularly when unusual histological features, such as sebaceous differentiation, are present.

Sebaceous Carcinoma Arising in a Precursor Benign Sebaceoma: Rare Example Highlighting the Utility of Immunohistochemistry Including p53

Abstract: Reports of sebaceous carcinoma arising from a pre-existing benign precursor are extremely sparse in the literature. We describe a case in which there was clear transition between sebaceoma and sebaceous carcinoma, with a different pattern of p53 staining in each component.

Functional divergence exists in mucilage-mediated seed dispersal, but not in germination of myxospermic Lepidium campestre and Lepidium draba (Brassicaceae)

The aim of this study was to compare the role of mucilage in seed dispersal and germination of the two closely related invasive weed species, Lepidium campestre and Lepidium draba. The study investigated the staining patterns of mucilage and its effects on seed germination, water uptake, dehydration, adherence to sand particles and dispersal by wind, water and animals following standard protocols. The mucilaginous seeds of L. campestre had a higher pectin and cellulose content than L. draba. This difference has a significant effect on water uptake and retention. The mucilage increased water uptake in both species, with L. campestre seeds absorbing significantly more water (836% compared to 75% in L. draba). Remarkably, the mucilage of both species had very little effect on seed germination. Significantly, the seeds of L. draba readily germinated under dry conditions, suggesting that this species may have developed different germination strategies. In addition, the seeds and fruits of L. draba showed a higher potential for dispersal via water and wind, whereas those of L. campestre adhered to animals. Further, L. campestre seeds showed significantly stronger adherence to sand particles (1872%) compared to those of L. draba (445%). This suggests that L. campestre seeds are more likely to disperse locally and possibly adhere to animals or soil particles. Our results suggest that mucilage produce by the two invasive weed species results in different methods of dispersal, which may help explain differences in their distribution patterns.

A Multi-task learning U-Net model for end-to-end HEp-2 cell image analysis

Antinuclear Antibody (ANA) testing is pivotal to help diagnose patients with a suspected autoimmune disease. The Indirect Immunofluorescence (IIF) microscopy performed with human epithelial type 2 (HEp-2) cells as the substrate is the reference method for ANA screening. It allows for the detection of antibodies binding to specific intracellular targets, resulting in various staining patterns that should be identified for diagnosis purposes. In recent years, there has been an increasing interest in devising deep learning methods for automated cell segmentation and classification of staining patterns, as well as for other tasks related to this diagnostic technique (such as intensity classification). However, little attention has been devoted to architectures aimed at simultaneously managing multiple interrelated tasks, via a shared representation.In this paper, we propose a deep neural network model that extends U-Net in a Multi-Task Learning (MTL) fashion, thus offering an end-to-end approach to tackle three fundamental tasks of the diagnostic procedure, i.e., HEp-2 cell specimen intensity classification, specimen segmentation, and pattern classification. The experiments were conducted on one of the largest publicly available datasets of HEp-2 images. The results showed that the proposed approach significantly outperformed the competing state-of-the-art methods for all the considered tasks.

Perfusion staining methods for visualization of the intact microvascular networks in whole mount skeletal muscle preparations.

Visualization of the intact microvascular network in skeletal muscle requires labeling the entire network in whole mount preparations where muscle fibre length can be set to near optimal but the tools to do this are not clear. We intravascularly injected CD-1 mice with different fluorescently labelled lectins (fluorescent isolectin GS-IB4 (ISO), wheat germ agglutinin (WGA), lycopersicon esculentum (tomato) (LYCO) or FITC-labelled gel. Soleus, extensor digitorum longus, diaphragm, gluteus maximus and cremaster muscles were excised, pinned at optimal sarcomere length and viewed using fluorescence microscopy. WGA and LYCO were the effective at labeling the entire vascular network with WGA labeling capillaries more bright. ISO labelled the arteriolar vasculature and early segments of the capillaries but not the full length of the capillaries or the venular network. FITC-labelled gel was effective at labelling the microvascular network but not all small vessels were consistently labelled. The pattern of staining for each labelling method was similar across all muscle fibre-types tested. WGA was optimal for perfusion labeling and visualization of the intact microvascular network in whole mount skeletal muscle preparations and can be used in combination with ISO to distinguish the arteriolar and venous sides of the network.

Intercellular, Linear Direct Immunofluorescence Staining Pattern of Sweat Glands and Hair Follicles may be Used as a Diagnostic Marker for Pemphigus Vulgaris in Cases where Epidermis is Missing.

Due to its rarity, it takes months for patients to be correctly diagnosed with pemphigus vulgaris. This delay can be exacerbated by factors such as incorrect biopsy site selection. When the epidermis detaches from the dermis, evaluating fluorescence patterns is impossible. This monocentric, retrospective study aimed to assess the clinical significance of the honeycomb-like staining pattern of adnexal structures in patients with pemphigus vulgaris. The study was conducted in Munich, from 1 January 2012 to 31 December 2020. Forty-four biopsy samples for direct immunofluorescence microscopy from pemphigus patients along with 44 samples from control patients were included. The fluorescence intensity of adnexal structures did not differ significantly from that of the epidermis in pemphigus, except for hair follicles and sweat glands with C3 staining, where the epidermis showed stronger fluorescence. The sensitivity for the characteristic fluorescence pattern of at least 1 adnexal structure was 88.64% (n = 39/44, 95% CI 75.44% to 96.21%), with 100% specificity. The positive predictive value of adnexal fluorescence was 100%, and the negative predictive value was 89.8%. It was found that the honeycomb-like staining pattern of sweat glands, sweat gland ducts, and hair follicles is a highly specific marker for pemphigus vulgaris.

EXTH-73. TARGETING THE CELL ADHESION CLUSTER OF DIFFERENTIATION 44 TRANSMEMBRANE GLYCOPROTEIN SENSITIZES CRANIOPHARYNGIOMAS TO IMMUNOTHERAPY

Abstract BACKGROUND Craniopharyngiomas are categorized as an adamantinomatous craniopharyngioma (ACP) or papillary craniopharyngioma (PCP) subtype with distinct histological, pathological and genetic differences. Studies propose the potential therapeutic benefit of targeting PD-L1 and/or PD-1 in both ACP and PCP subtypes using immune checkpoint inhibitors (ICI). The alternative mRNA splice variant of CD44, CD44v9, is expressed on cancer stem cells and plays multiple key roles including drug resistance and tumor cell invasiveness. CD44 positively regulates PD-L1, but the role of this transmembrane glycoprotein in craniopharyngioma tumorigenesis is unknown. Objectives: To replicate the complexity of the craniopharyngioma tumor microenvironment (TME) by developing a novel patient tissue-generated craniopharyngioma in vitro organoid preclinical model for the development of therapeutic strategies. Approach: NanoString CosMx™ Spatial Molecular Imager (SMI) was performed using FFPE tissue sections prepared from patient craniopharyngiomas. Quantification of PD-L1, and stem cell transmembrane glycoproteins CD44v9 and TROP2 was performed using multiplex immunofluorescence (MxIF). Craniopharyngioma patient-derived organoids (CranioPDOs) were generated and co-cultured with autologous immune cells (IMM) generated from patient peripheral blood mononuclear cells. CranioPDO/IMM co-cultures were used to predict the potential therapeutic benefit in response to tyrosine kinase inhibitor cabozantinib (Cabo) plus pembrolizumab (Pembro) or atezolizumab (Atezo) combinatorial therapy. RESULTS SMI analysis showed expression of CD44 and TROP2 positive cells, cancer-associated fibroblasts and myeloid-derived suppressor cells (MDSCs) consistent with the staining pattern of PD-L1. MxIF of matched tissues showed expression of CD44v9 and TROP2 within the same CD44 positive spatially mapped cell populations. Combinatorial treatment of CranioPDO/IMM co-cultures with Cabo plus Pembro or Atezo led to depletion of MDSCs correlating with decreased expression of CD44v9 and a significant increase in cytotoxic T lymphocyte proliferation and tumor cell death. CONCLUSION Our preclinical in vitro organoid data support the potential therapeutic benefit of ICI in combination with cabozantinib for the treatment of craniopharyngiomas.

TMOD-06. ESTABLISHMENT OF PITUITARY NEUROENDOCRINE TUMOR (PITNET) ORGANOID MODELS

Abstract Pituitary neuroendocrine tumors (PitNETs) are tumors that originate from the anterior pituitary gland. While they are mostly benign, they can cause various clinical symptoms due to hormonal secretion abnormalities. PitNET can be broadly categorized into three lineages, each of which has its differentiation controlled by specific transcription factors. Treatment options for PitNET include surgical resection and pharmacotherapy, but available drugs are limited, and their efficacy markers remain unclear. To develop novel drugs for PitNET and identify their efficacy markers, it is necessary to develop new research models. In recent years, organoid models have garnered attention as novel research models for various cancer types. In this study, we established novel organoid models derived from PitNET patients and analyzed their histological and endocrinological characteristics. Surgical specimens from 4 cases of somatotroph adenoma, 1 case of double adenoma, 1 case of gonadotroph adenoma, and 1 case of corticotroph adenoma were minced in dissection medium and embedded in Matrigel basement membrane matrix respectively. The organoids were cultured under 5% CO2 at 37°C using media supplemented with various growth factors. After 28 days of culture, formalin-fixed paraffin-embedded sections were prepared to examine histological homology with patient tumors. Hematoxylin and eosin staining results showed that all organoids maintained cell morphology and tissue structure similar to the original tumors. Immunohistochemical staining for markers specific to each PitNET revealed similar staining patterns between the organoids and the original tumors. Furthermore, analysis of hormone levels in the culture medium showed that hormone production capabilities, such as growth hormone and follicle stimulating hormone, were maintained even after long-term culture. In this study, we successfully established PitNET organoid models that retain histological and endocrinological characteristics. These models are expected to serve as useful research tools for elucidating the molecular mechanisms of PitNET and developing novel therapies targeting critical molecular abnormalities.

Assessing expression patterns of PTGR1, a potential biomarker for acylfulven sensitivity in urothelial carcinoma

BackgroundMetastatic urothelial carcinoma (mUC) has a poor prognosis, despite recent therapeutic advancements. Prostaglandin Reductase 1 (PTGR1) is essential for activating acylfulvens, a promising class of drugs for treating a subset of urothelial carcinoma (UC) patients. The efficacy of acylfulvens depends on PTGR1 activity and defects in the nucleotide excision repair (NER) pathway, notably ERCC2 mutations present in 10–15% of bladder tumors. This study identifies patients eligible to be included in acylfulven clinical trials based on the presence of PTGR-1 by immunohistochemistry (IHC) staining, RNA expression level and mutations in the NER pathway. Additionally, it evaluates PTGR-1 expression as a prognostic biomarker.MethodsThree PTGR-1 antibodies were tested in a kidney cancer cell line A498 and in tissues with known PTGR1 expression (liver, tonsils, pancreas, small intestine, etc.). Patients with untreated mUC receiving 1st line platinum from Dec. 2019 to Dec. 2021 in the Capital Region, Denmark, were included retrospectively. FFPE tumor samples were collected, and tissue microarrays (TMAs) were constructed. TMAs were stained with the best-performing PTGR1 antibody, RNA expression was analyzed using Nanostring nCounter PanCancer panel and gene mutations were assessed using a targeted genomic panel (TSO-500). Kaplan-Meier and multivariate Cox regression assessed overall survival and covariate impacts.ResultsThe AB181131 PTGR1 antibody was the most reliable in validation tissues. Tumors from 71 mUC patients were used to construct the TMA, and 40% of tumors scored positive for PTGR1 by IHC staining. A normalized PTGR1 RNA cutoff at 2,550 normalized counts achieved an AUC of 0.9, defining 35 samples as positive with a sensitivity of 96% and specificity of 85% in relation to IHC-positivity. Differential expression showed a significant upregulation of PTGR1 RNA in PTGR1 IHC-positive cases. NER-deficiency and PTGR1 positivity (mutations in ERCC1, ERCC2, ERCC3, ERCC4) was seen in 9 patients (13%). Median overall survival was 16 months in the cohort. Overall survival (OS) analysis indicates that overexpression of PTGR1 RNA was associated with a reduced median OS (12 months vs. 25 months, p = 0.039, log-rank).ConclusionPTGR1 IHC staining pattern using the Abcam AB181131 antibody is highly correlated with PTGR1 RNA expression. 13% in our cohort were identified as NER deficient and PTGR1 positive. Lower levels of PTGR1 indicates better outcome in this cohort.

Distinct Chemical Determinants are Essential for Achieving Ligands for Superior Optical Detection of Specific Amyloid-β Deposits in Alzheimer's Disease.

Aggregated forms of different proteins are common hallmarks for several neurodegenerative diseases, including Alzheimer's disease, and ligands that selectively detect specific protein aggregates are vital. Herein, we investigate the molecular requirements of thiophene-vinyl-benzothiazole based ligands to detect a specific type of Aβ deposits found in individuals with dominantly inherited Alzheimer's disease caused by the Arctic APP E693G mutation. The staining of these Aβ deposits was alternated when switching the terminal heterocyclic moiety attached to the thiophene-vinyl-benzothiazole scaffold. The most prevalent staining was observed for ligands having a terminal 3-methyl-1H-indazole moiety or a terminal 1,2-dimethoxybenzene moiety, verifying that specific molecular interactions between these ligands and the aggregates were necessary. The synthesis of additional thiophene-vinyl-benzothiazole ligands aided in pinpointing additional crucial chemical determinants, such as positioning of nitrogen atoms and methyl substituents, for achieving optimal staining of Aβ aggregates. When combining the optimized thiophene-vinyl-benzothiazole based ligands with a conventional ligand, CN-PiB, distinct staining patterns were observed for sporadic Alzheimer's disease versus dominantly inherited Alzheimer's disease caused by the Arctic APP E693G mutation. Our findings provide chemical insights for developing novel ligands that allow for a more precise assignment of Aβ deposits, and might also aid in creating novel agents for clinical imaging of distinct Aβ aggregates in AD.

Analysis of antinuclear antibody in 9 528 pregnant women during early pregnancy in a hospital in Qingdao City

To analyze the positivity rate and titer of antinuclear antibody (ANA), as well as nuclear pattern and target antigen of ANA in healthy pregnant women during early pregnancy in Qingdao area. A prospective cohort study design was used to include a total of 9 528 healthy pregnant women registered at the Women and Children's Hospital Affiliated to Qingdao University from March 2023 to June 2024.Indirect immunofluorescence assay (IIF) was used to detect ANA, its titer and cell staining pattern. Fifteen specific antibodies were tested using the magnetic bar code immunofluorescent luminescence method. Logistic regression model was used to analyze the risk factors of pregnancy with autoimmune disease(AID). The results showed that among 9 528 pregnant women in early pregnancy, 1 346 cases (14.1%) were positive of ANA, including 1 011 cases with a titer of 1∶100 (10.6%), 236 cases (2.5%) with a titer of 1∶320, and 99 cases (1.0%) were detected at a titer >1∶320. Among the 1 346 ANA-positive pregnant women, nuclear granular type accounted for the highest proportion (483 cases, 35.9%), followed by speckled type (347 cases, 25.8%) and cytoplasmic type (176 cases, 13.1%).Then, pregnant women with ANA titers ≥1∶100 were detected 15 specific antibodies.Anti-SSA was tested in 121 cases accounted for the majority, followed by 110 cases with anti-Ro-52, 56 cases with anti-SSB, 51 cases with anti-mitochondrial M2 subtype antibodies and 37 cases with anti-centromere B. In conclusion,in healthy pregnant women in Qingdao area, ANA positivity rate was 14.1%, and the titer of ANA was mainly at 1∶100.The predominant nuclear patterns were nuclear granular and speckled types.The specific autoantibodies were mainly anti-SSA antibodies and anti-Ro-52 antibodies.The detection of ANA and specific autoantibodies is of great significance for early prediction, diagnosis, and intervention of autoimmune diseases during pregnancy.

Current status and challenges in HER2 IHC assessment: scoring survey results in Japan.

This study aimed to assess the concordance of human epidermal growth factor receptor 2 (HER2) expression scoring by immunohistochemistry (IHC) among practicing pathologists in Japan, given the challenging nature of scoring and the critical role of HER2 status in breast cancer management. Whole slide images (WSI) from 20 invasive breast cancer cases (1 representative WSI per case) selected to represent a diverse IHC scores and staining patterns were used in an online survey involving seven reference pathologists who established consensus HER2 IHC scores (0 to 3 +) decided by majority interpretation. Participating pathologists nationwide scored the same 20 WSI cases online using the American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) 2018 guidelines. Deidentified case metadata were registered in the uPath system. A total of 144 participating pathologists responded. The scoring results of the participating pathologists most commonly agreed with the consensus IHC score, followed by a ± 1 point deviation and no survey responses with > 1 point deviation. The mean percentage of agreement with the consensus score for all 20 cases was 63.4%. In cases where the reference pathologists' scores were discordant, the participating pathologists also showed a lower concordance rate. This study highlighted the current status of HER2 expression scoring by IHC for breast cancer among pathologists in Japan. These findings underscore the challenges in HER2 IHC scoring cases and emphasize the need for improved standardization and training, especially in the evolving landscape of HER2-targeted therapies.

Subcellular expression of CD30 in cutaneous mastocytosis-An important factor for targeted treatment.

The subcellular distribution of CD30 on mast cells and the presence of eosinophils in cutaneous mastocytosis require further investigation, especially as the cell surface expression of CD30 is critical for the therapeutic response of systemic mastocytosis to brentuximab vedotin. Investigation of 147 biopsy specimens from 143 patients with cutaneous mastocytosis for mast cell density and distribution, frequency of CD30 expression, CD30 staining patterns, and presence and distribution of eosinophils. Correlation with clinical patterns. Retrospective multicenter immunohistochemical study of CD30 expression, eosinophils and basic clinical data in cutaneous mastocytosis. CD30 expression was found in all samples (cut-off: ≥1%), whereby the staining was predominantly cytoplasmic in 99% of the samples. Additional membrane staining was detected in 62% of the samples. Surface expression of CD30 was more common in biopsy specimens with a high mast cell burden and in biopsy specimens with a higher CD30 expression rate. Eosinophils were admixed in 58% of the samples. Females and older patients showed a trend of a lower mast cell burden. Retrospective study on formalin-fixed and paraffin-embedded tissue without functional analysis. Most cases of cutaneous mastocytosis show cell surface expression of CD30 expression and is, therefore, in principle, accessible for therapy with antibodies against CD30, provided the overall situation of the patient warrants.