Development of an Artificial-Intelligence-Based Tool for Automated Assessment of Cellularity in Bone Marrow Biopsies in Ph-Negative Myeloproliferative Neoplasms.

The cellularity assessment in bone marrow biopsies (BMBs) for the diagnosis of Philadelphia chromosome (Ph)-negative myeloproliferative neoplasms (MPNs) is a key diagnostic feature and is usually performed by the human eyes through an optical microscope with consequent inter-observer and intra-observer variability. Thus, the use of an automated tool may reduce variability, improving the uniformity of the evaluation. The aim of this work is to develop an accurate AI-based tool for the automated quantification of cellularity in BMB histology. A total of 55 BMB histological slides, diagnosed as Ph- MPN between January 2018 and June 2023 from the archives of the Pathology Unit of University "Luigi Vanvitelli" in Naples (Italy), were scanned on Ventana DP200 or Epredia P1000 and exported as whole-slide images (WSIs). Fifteen BMBs were randomly selected to obtain a training set of AI-based tools. An expert pathologist and a trained resident performed annotations of hematopoietic tissue and adipose tissue, and annotations were exported as .tiff images and .png labels with two colors (black for hematopoietic tissue and yellow for adipose tissue). Subsequently, we developed a semantic segmentation model for hematopoietic tissue and adipose tissue. The remaining 40 BMBs were used for model verification. The performance of our model was compared with an evaluation of the cellularity of five expert hematopathologists and three trainees; we obtained an optimal concordance between our model and the expert pathologists' evaluation, with poorer concordance for trainees. There were no significant differences in cellularity assessments between two different scanners.

Combined fine needle aspiration cytology and core needle biopsy in the same setting: A two-years' experience.

INTRODUCTION: Fine needle aspiration cytology (FNAC) combined with rapid on-site evaluation (ROSE) and ancillary techniques is an accurate diagnostic tool for many pathologies. However, in some cases, it may not be sufficient for actionable diagnoses or molecular testing, especially for cases that require large immunohistochemical panels or cases in which histological features are mandatory for the diagnosis. Core needle biopsy (CNB), on the contrary, provides samples that are suitable for histological features and sufficient for all ancillary studies. However, CNB is often performed by radiologists or clinicians without the direct participation of cytopathologists, which can lead to missed or delayed diagnoses. This study reports on the experience of combining FNAC and CNB performed in one setting by cytopathologists. The aim was to evaluate the impact of CNB on FNAC and the diagnostic efficiency of the combined procedures. MATERIALS AND METHODS: One hundred forty-two FNAC and CNB procedures performed in the same setting over a period of 2 years were analysed. The FNAC diagnoses were compared and integrated with the subsequent CNB diagnoses. The impact of CNB was categorized as follows: non-contributory, in cases of inadequate samples; confirmed, when the CNB and FNAC diagnoses were the same; improved, when the CNB diagnosis was consistent with the FNAC diagnosis and further specified the corresponding entity; allowed, when CNB produced a diagnosis that could not be reached by FNAC; changed, when the CNB changed the previous FNAC diagnosis. RESULTS: CNB confirmed the FNAC diagnosis in 40.1% of cases (n = 57/142). CNB improved the FNAC diagnosis in 47.2% of cases (n = 67/142). CNB allowed a diagnosis that could not be performed on FNAC in 2.1% of cases (n = 3/142). CNB changed a previous FNAC diagnosis in 2.1% of cases (n = 3/142). CNB was non-contributory in 8.4% of cases (n = 12/142). CNB produced a positive impact on the whole diagnostic procedure in 51.4% of total cases (n = 73/142). The combined FNAC and CNB resulted in actionable diagnoses in 91.5% of all cases (n = 130/142). A complete molecular assessment was successfully performed in 14.7% of cases (n = 21/142) utilizing either FNAC or CNB material. CONCLUSIONS: The combined use of FNAC and CNB in one setting improves the diagnostic accuracy of both procedures. This approach exploits the advantages of each procedure, enhancing the accuracy of the final diagnosis.

Changes in mucosal IgG4+- and IL-10+-cell frequencies in adults with eosinophilic esophagitis on a two-food elimination diet.

Eosinophilic esophagitis (EoE) is increasingly diagnosed in patients with dysphagia. Type-2 immunity can induce EoE histopathology via non-IgE-dependent mechanisms, possibly involving IgG4 and IL-10. To elucidate the contribution of this response to EoE pathogenesis, we examined its association with clinical and histologic endpoints in adult EoE patients given a two-food elimination diet. IgG4- and IL-10-expressing cells were counted in esophageal biopsies and serum food-specific IgG4 measured at baseline and follow-up. Variables were correlated with histologic measures of disease activity. Patients exhibited significant reduction in esophageal eosinophilia and overall histology. A significant decrease in IL-10+-cell frequencies correlated with histologic changes. In contrast, a decline in serum and esophageal IgG4, while substantial, did not correlate with IL-10+-cell frequencies or histologic parameters. These results suggest a critical role of IL-10 in EoE pathogenesis. Conversely, IgG4 expression, while reflecting exposure to food antigens, is not obviously related to EoE histopathology or IL-10 expression.

Digital Examination of LYmph node CYtopathology Using the Sydney system (DELYCYUS): An international, multi-institutional study.

BACKGROUND: After a series of standardized reporting systems in cytopathology, the Sydney system was recently introduced to address the need for reproducibility and standardization in lymph node cytopathology. Since then, the risk of malignancy for the categories of the Sydney system has been explored by several studies, but no studies have yet examined the interobserver reproducibility of the Sydney system. METHODS: The authors assessed interobserver reproducibility of the Sydney system on 85 lymph node fine-needle aspiration cytology cases reviewed by 15 cytopathologists from 12 institutions in eight different countries, resulting in 1275 diagnoses. In total, 186 slides stained with Diff-Quik, Papanicolaou, and immunocytochemistry were scanned. A subset of the cases included clinical data and results from ultrasound examinations, flow cytometry immunophenotyping, and fluorescence in situ hybridization analysis. The study participants assessed the cases digitally using whole-slide images. RESULTS: Overall, the authors observed an almost perfect agreement of cytopathologists with the ground truth (median weighted Cohen κ = 0.887; interquartile range, κ = 0.210) and moderate overall interobserver concordance (Fleiss κ = 0.476). There was substantial agreement for the inadequate and malignant categories (κ = 0.794 and κ = 0.729, respectively), moderate agreement for the benign category (κ = 0.490), and very slight agreement for the suspicious (κ = 0.104) and atypical (κ = 0.075) categories. CONCLUSIONS: The Sydney system for reporting lymph node cytopathology shows adequate interobserver concordance. Digital microscopy is an adequate means to assess lymph node cytopathology specimens.

Prostatic stromal sarcoma: Report of a rare case in a young male and review of the literature.

INTRODUCTION AND IMPORTANCE: Common prostatic neoplasms are diagnosed frequently, whereas rare entities require particular awareness and special clinical management. CASE PRESENTATION: A 31-year-old man presented with dysuria, hematuria and urinary retention. Histomorphological analysis of material obtained by transurethral resection of the prostate initially favored a sarcomatoid carcinoma, but immunohistochemistry allowed the correct diagnosis of sarcoma of the specialized stroma of the prostate. CLINICAL DISCUSSION: The patient refused surgical treatment and, despite chemotherapy, he died 8 months after the diagnosis. Herein, we will highlight the diagnostic and therapeutic challenge of prostatic stromal sarcoma by illustrating this case and reviewing the relevant literature. CONCLUSION: Although rare and shadowed by more common neoplasms that may mimic it, prostatic stromal sarcoma should be considered in the differential diagnosis of bladder and prostate neoplasms because of its dismal prognosis.

Non-urothelial lesions of the urinary bladder A 14.5-year, single-institution review.

CONTEXT: In contrast to urothelial cancers, non-urothelial neoplasms involving the bladder are uncommon and often diagnostically challenging. These lesions include a variety of benign and malignant tumors often presenting with a combination of hematuria and the presence of a polypoid lesion at cystoscopy that may lead to an erroneous diagnosis of urothelial cancer. OBJECTIVE: We set out to quantify and classify the spectrum of non-urothelial lesions diagnosed in our institution, and briefly review the relevant literature on each lesion, with a focus on differential diagnosis and potential pitfalls. DESIGN: We performed a retrospective review (Jan 2008 - Jun 2022) of the cases diagnosed on TURB material at our institution. RESULTS: Out of 4071 TURB specimens, a total of 66 (1.62 %) non-urothelial lesions were identified. Most of these lesions were malignant (n = 51, 77 %), with metastases being the most common (n = 40, 60.6 %), followed by non-Hodgkin lymphoma (n = 8, 12 %). The remaining cases were benign lesions (n = 15, 22.7 %), with the most common being inflammatory myofibroblastic tumor (n = 4, 6.1 %) and endometriosis (n = 3, 4.5 %). CONCLUSIONS: In this retrospective case series, we identified various malignant and benign entities, some of which have been rarely reported in the bladder, such as paragangliomas, inflammatory myofibroblastic tumor, and leiomyosarcoma. These lesions may macroscopically and histologically mimic urothelial carcinoma. Because of their relative rarity and diagnostic overlap with conventional urothelial tumors, the pathologist should always keep in the mind the possibility of non-urothelial lesions.

Immunotherapy in Penile Squamous Cell Carcinoma: Present or Future? Multi-Target Analysis of Programmed Cell Death Ligand 1 Expression and Microsatellite Instability.

Background: Penile cancer (PC) is an extremely rare malignancy, and the patients at advanced stages have currently limited treatment options with disappointing results. Immune checkpoint inhibitors anti-programmed cell death 1 (PD-1)/programmed cell death ligand 1 (PD-L1) are currently changing the treatment of several tumors. Furthermore, the microsatellite instability (MSI) and the deficient mismatch repair system (dMMR) proteins represent predictive biomarkers for response to immune checkpoint therapy. Until present, few data have been reported related to PD-L1 expression and MSI in PC. The main aim of our study was the evaluation of PD-L1 expression in tumor cells (TCs) and tumor-infiltrating lymphocytes (TILs) in immune cells and the analysis of dMMR/MSI status in a large series of PCs. Methods: A series of 72 PC, including 65 usual squamous cell carcinoma (USCC), 1 verrucous, 4 basaloid, 1 warty, and 1 mixed (warty-basaloid), was collected. Immunohistochemistry (IHC) was performed to assess PD-L1 expression using two different anti-PD-L1 antibodies (clone SP263 and SP142 Ventana) and MMR proteins expression using anti-MLH1, anti-PMS2, anti-MSH2, and anti-MSH6 antibodies. PCR analysis was performed for the detection of MSI status. Results: Of the 72 PC cases analyzed by IHC, 45 (62.5%) cases were TC positive and 57 (79%) cases were combined positive score (CPS) using PDL1 SP263. In our cohort, TILs were present in 62 out of 72 cases (86.1%), 47 (75.8%) out of 62 cases showed positivity to PDL1 clone SP142. In our series, 59 cases (82%) had pMMR, 12 cases (16.7%) had lo-paMMR, and only 1 case (1.3%) had MMR. PCR results showed that only one case lo-paMMR was MSI-H, and the case dMMR by IHC not confirmed MSI status. Conclusion: Our findings showed that PD-L1 expression and MSI status represent frequent biological events in this tumor suggesting a rationale for a new frontier in the treatment of patients with PC based on the immune checkpoint inhibitors.

Hematuria in breast cancer: don't forget bladder metastases!

The bladder is a rare site for breast cancer metastases, and only occasional reports are present in the literature. Most cases coexist with synchronous metastases elsewhere, but isolated cases of a single metastatic localization in the urinary bladder have been reported. The most common symptoms of a metastatic localization of breast cancer to the urinary bladder are hematuria and voiding dysfunction. Herein we present three cases of urinary bladder metastasis from breast carcinoma, all presenting with gross hematuria as the only symptom. After a review of the relevant literature, we discuss the clinical and histological characteristics unique to our cases, highlighting potential clinical and pathological diagnostic pitfalls and differential diagnoses.

Cytological Diagnosis of Aggressive Small-Cell Lymphomas.

BACKGROUND: Despite their sometimes deceivingly bland appearance, some small-cell lymphomas are very aggressive and the prognosis of patients depends on a prompt diagnosis that allows the initiation of appropriate therapy. SUMMARY: The present review discusses the salient cytological features of the most common aggressive small-cell lymphomas and then proceeds to analyze their main diagnostic criteria, including the usage of ancillary techniques. KEY MESSAGES: Lymph node fine-needle aspiration cytology is a fast, safe, cheap, minimally invasive, and accurate procedure that can be used for a prompt and accurate diagnosis of lymphomas.

Real-world experience with the Sydney System on 1458 cases of lymph node fine needle aspiration cytology.

INTRODUCTION: Lymph node (LN) fine needle aspiration cytology (FNAC) is a safe, quick, inexpensive, reliable, and minimally invasive technique for the diagnosis of lymphadenopathies. Recently, an international committee of experts proposed guidelines for the performance, classification, and reporting of LN-FNAC: the Sydney System. We set out to analyse the diagnostic performance of the Sydney System in a retrospective study. METHODS: We retrieved 1458 LN-FNACs, reformulated the diagnoses according to the Sydney System, and compared them to the histological control where available (n = 551, 37.8%). RESULTS: The risk of malignancy for each of the five categories was 66.7% for inadequate/insufficient, 9.38% for benign (overall: 0.84%), 28.6% for atypical, 100% for suspicious and 99.8% for malignant. LN-FNAC showed a sensitivity of 97.94%, a specificity of 96.92%, a positive predictive value of 99.58%, and a negative predictive value of 86.30%. CONCLUSIONS: These data support the usage of LN-FNAC as an agile first-level technique in the diagnosis of lymphadenopathies. The Sydney System supports and enhances this role of LN-FNAC, and its adoption is encouraged. In negative cases, coupled with ancillary techniques, LN-FNAC can reassure the clinician regarding the benignity of a lymphadenopathy and indicate the need for clinical follow-up, which will catch possible false negatives. In positive cases, LN-FNAC can provide sufficient information, including predictive biomarkers, to initiate management and obviate the need for subsequent, more invasive procedures. Given its speed, minimal invasiveness, and low cost, LN-FNAC can be performed in most cases, even when more invasive techniques are not feasible.

The best prostate biopsy sampling system-fusion and systematic biopsy: A single center experience.

BACKGROUND: Prostate cancer is the second most commonly diagnosed cancer in men. The diagnostic accuracy in prostate cancer can be increased by employing a preliminary multiparametric MRI followed by a fusion-targeted biopsy. METHODS: To compare the diagnostic accuracy of fusion-targeted biopsy with the standard systematic biopsy in prostate cancer patients, we enrolled 139 patients on which we performed 139 prostate biopsies consisting of three targeted samples followed by 12 regular systematic samples. Based on histology, we analyzed the diagnostic performance of the two methods. RESULTS: Both methods were equally good at detecting clinically significant cancer (83.3%, 50/60), while systematic biopsy detected more clinically insignificant cancers. However, the best diagnostic performance is obtained by combining the two methods. CONCLUSION: The two methods are best seen as synergistic, and the addition of fusion biopsy can be used to detect more clinically significant prostate cancers than systematic biopsy alone.

Malignant melanoma of the prostate gland: A systematic review.

AIMS: Prostatic melanoma is a rare malignancy about which only scattered case reports and no systematic reviews have been published to date. We sought to better inform clinicians and pathologists caring for these patients by gathering all available evidence on the topic. METHODS: We performed a systematic review of English and non-English articles indexed in PubMed, EMBASE, Scopus and Google Scholar about primary and metastatic prostatic melanoma. RESULTS: In total, 25 studies describing 45 cases were identified. Most cases were metastases to the prostate, with only 10 primary prostatic cases. The median age of patients was 61 years with a wide range, and 89% were symptomatic at presentation, most commonly with obstructive symptoms (83%). Diagnosis requires histopathological analysis and often immunohistochemistry. Metastatic melanoma in the prostate carries a dismal prognosis with median overall survival of 3 months; on the other hand, among primary prostatic melanomas reported in the literature, 29% survived longer than 5 years. The most reasonable therapeutic approach consists in radical surgery possibly followed by adjuvant therapy.

Advanced non-small cell lung cancer: Rapid evaluation of EGFR status on fine-needle cytology samples using Idylla.

Advanced non-small cell lung cancer (NSCLC) needs to be managed rapidly; therefore, a rapid assessment of the epidermal growth factor receptor (EGFR) status is mandatory. Computed Tomography (CT)-guided or Ultrasound (US)-guided Fine-Needle Aspiration Cytology (FNAC) allows a rapid diagnosis of both primary and metastatic tumor through rapid on-site evaluation (ROSE) and the proper management of diagnostic material. Idylla (Biocartis, Mechelen, Belgium) is an automated RT-PCR system which evaluates the mutational status of specific genes in less than two hours. In this study, the EGFR mutational status in advanced NSCLC was analyzed on 28 FNAC samples with Idylla. After ROSE, residual FNAC material and/or additional passes were pipetted into the Idylla EGFR cartridge. Patients endorsed a consent form before carrying out the analysis. Results were controlled by pyrosequencing. Adequate EGFR status was obtained in 26/28 cases (22 wild type and 4 mutated). Mutated cases harbored EGFR Exon 19 deletion and L858R point mutation. In 2/28 cases the analysis failed. The combination of FNAC, ROSE and Idylla is a rapid, accurate and effective method that can be conveniently used to assess EGFR status in advanced NSCLC.

Testing EGFR with Idylla on Cytological Specimens of Lung Cancer: A Review.

The current standard of care for advanced non-small-cell lung cancer is based on detecting actionable mutations that can benefit from targeted therapy. Comprehensive genetic tests can have long turn-around times, and because EGFR mutations are the most prevalent actionable mutation, a quick detection would enable a prompt initiation of targeted therapy. Furthermore, the scarcity of diagnostic material means that sometimes only cytologic material is available. The Idylla™ EGFR assay is a real-time PCR-based method able to detect 51 EGFR mutations in 2.5 h. Idylla is validated for use only on FFPE sections, but some researchers described their experiences with cytological material. We reviewed the relevant literature, finding four articles describing 471 cases and many types of cytological input material: smears, cell-block sections, suspensions, and extracted DNA. The sensitivity, specificity, and limit of detection appear comparable to those obtained with histological input material, with one exception: the usage of scraped stained smears as input may reduce the accuracy of the test. In conclusion, usage of cytological material as input to the Idylla EGFR test is possible. A workflow where common mutations are tested first and fast, leaving rarer mutations for subsequent comprehensive profiling, seems the most effective approach.

Nervous Nipple: Pseudopolythelia Caused by a Neurofibroma of the Areola.

ABSTRACT: The presence of a supernumerary nipple inside the original areola is a rare condition termed intra-areolar polythelia. Rarely, a lesion can macroscopically resemble a nipple. We report a case of a solitary neurofibroma (by itself rare in the areola) mimicking a second, twin nipple. In this case, these 2 rare conditions merge resulting in pseudopolythelia. The relevant literature on polythelia and neurofibromas of the breast is briefly reviewed.

Ki67 in Gleason Pattern 3 as a Marker of the Presence of Higher-Grade Prostate Cancer.

PURPOSE: Prostate biopsies may undergrade up to half of all prostate cancers (PCs), delaying definitive treatment by up to 3 years. One cause of undergrading is the partial sampling inherent in the technique. Because of this, a prostate biopsy that appears to be Gleason 3+3=6 may come either from a true 3+3=6 tumor or from a higher-grade tumor that has been sampled only partially. The main goal of the present study is to identify a way to distinguish these 2 kinds of "Gleason 3+3=6" biopsies.Mounting evidence hints at the possibility that Gleason pattern 3 associated with higher-grade PC (aG3) is biologically distinct from pure Gleason pattern 3 (pG3). MATERIALS AND METHODS: In this study, we used immunohistochemistry and computer-aided image analysis to compare the expression of Ki67, cyclin D1, MYC, and p53 between foci of aG3 and pG3, to search for a marker that could distinguish them. RESULTS: The expression of Ki67 differed significantly between pG3 and aG3. The average Ki67 labeling index was 1.63% for pG3 and 7.62% for aG3 (P<0.01); the average number of Ki67+ cells per high-power field was 17 for pG3 and 60 for aG3 (P<0.01). The other markers did not differ significantly between pG3 and aG3. CONCLUSIONS: When a biopsy only shows Gleason pattern 3 PC, Ki67 immunohistochemistry could be used to distinguish the nodules of true Gleason score 3+3=6 from those that only appear to be 3+3=6 because of a sampling error. This would dramatically improve the diagnostic performance of prostate biopsies and the management of early PC.

A Rare Case of Castleman Disease Presenting as an Ovarian Tumor.

Castleman disease (CD) is a rare lymphoproliferative disorder of unknown etiology. Its most common location is the mediastinum, but many other sites have been reported. We report a case of primary CD of the ovary, a rare localization with only 2 cases including the present case described in the world literature to date. A 58-yr-old woman who initially presented with abdominal pain underwent computed tomography scan which showed bilateral well-circumscribed solid adnexal masses. Because an ovarian bilateral tumor was suspected the patient was treated with a hysterectomy and bilateral salpingo-oophorectomy and the histopathologic examination confirmed the diagnosis of CD hyaline-vascular type of the right ovary associated with a contralateral fibroma. Three years after surgery the patient is alive and well and shows no signs of recurrent disease. The occurrence of this rare presentation of CD is the subject of this report. The problems of differential diagnosis with the most frequent lesions of the female pelvis are also discussed.

Fatal Outcome Consequent to an Endoscopic Full Thickness Resection of a Colonic Lateral Spreading Tumor: A Case Report.

BACKGROUND Endoscopic full-thickness resection represents an innovative procedure, used in selected patients that allows lesions en-bloc resection with an integral wall specimen available for histopathological definition. Bleeding and perforation are known to be the most frequent procedure-related adverse events. We report a case of entero-colonic fistula as complication of an endoscopic full-thickness resection. CASE REPORT A 77-year-old male, with a personal history of right-hemicolectomy for a colonic adenocarcinoma presented to our department for a routine colonoscopy that showed the presence of a 25 mm lateral spreading tumor localized at about 50 cm from the anal margin. A full-thickness resection of the lateral spreading tumor using the over-the-scope clip device was performed. After 4 weeks, because of abdominal pain, weight loss, diarrhea, and signs of malnutrition, the patient underwent a new colonoscopy showing hyperemic mucosa with ulcerations in all colonic segments and, at the site of the previous endoscopic full-thickness resection, an orifice of an entero-colonic fistula. The histological definition was suggestive for ulcerative proctocolitis and confirmed the presence of small bowel mucosa at fistula orifice. An intussusception at the level of fistula with consequent intestinal obstruction caused a worsening of clinical conditions and finally the patient death for a septic peritonitis. CONCLUSIONS Full thickness resection represents an innovative tool for en-bloc resection of gastrointestinal tumoral lesion, but procedural complications and limitations must be considered before performing this procedure.