Use of Novel Open-Source Deep Learning Platform for Quantification of Ki-67 in Neuroendocrine Tumors - Analytical Validation.

Background: Neuroendocrine tumors (NETs) represent a diverse group of neoplasms that arise from neuroendocrine cells, with Ki-67 immunostaining serving as a crucial biomarker for assessing tumor proliferation and prognosis. Accurate and reliable quantification of Ki-67 labeling index is essential for effective clinical management. Methods: We aimed to evaluate the performance of open-source/open-access deep learning cloud-native platform, DeepLIIF (https://deepliif.org), for the quantification of Ki-67 expression in gastrointestinal neuroendocrine tumors and compare it with the manual quantification method. Results: Our results demonstrate that the DeepLIIF quantification of Ki-67 in NETs achieves a high degree of accuracy with an intraclass correlation coefficient (ICC) = 0.885 with 95% CI (0.848-0.916) which indicates good reliability when compared to manual assessments by experienced pathologists. DeepLIIF exhibits excellent intra- and inter-observer agreement and ensures consistency in Ki-67 scoring. Additionally, DeepLIIF significantly reduces analysis time, making it a valuable tool for high-throughput clinical settings. Conclusion: This study showcases the potential of open-source/open-access user-friendly deep learning platforms, such as DeepLIIF, for the quantification of Ki-67 in neuroendocrine tumors. The analytical validation presented here establishes the reliability and robustness of this innovative method, paving the way for its integration into routine clinical practice. Accurate and efficient Ki-67 assessment is paramount for risk stratification and treatment decisions in NETs and AI offers a promising solution for enhancing diagnostic accuracy and patient care in the field of neuroendocrine oncology.

Morphological Spectrum of Vesiculobullous Skin Lesions: An Institutional Perspective.

Introduction A vesiculobullous lesion of the skin encompasses a group of dermatological disorders with protean clinicopathological features. They usually occur as a part of the spectrum of various infectious, inflammatory, drug-induced, genetic, and autoimmune disorders. Therefore, accurate diagnosis of these lesions is essential for appropriate management and to reduce the associated morbidity and mortality. The conventional skin punch biopsy is the mainstay in the diagnosis of dermatological diseases, especially when combined with confirmatory tests, such as direct immunofluorescence (DIF). Our study evaluated the clinicopathological spectrum of vesiculobullous lesions. Methods We studied 150 cases of vesiculobullous lesions at the Department of Histopathology, Liaquat National Hospital and Medical College Karachi, Pakistan. Written and informed consent was taken from the patients followed by skin punch procedure in which three biopsies were obtained, which included one biopsy from the lesion and two peri-lesional biopsies. One peri-lesional biopsy was sent in cryomatrix for DIF studies, whereas the other two were sent in formalin to follow the standard tissue-processing protocol. Results Our results showed that most patients belonged to the geriatric age group of more than 50 years (44.7%), and 54.7% of the patients were females. Total 74.7% of the patients had generalized lesions, followed by lower limbs (9.3%) and trunk (7.3%) involvement. Most patients were diagnosed with bullous pemphigoid (31.3%), followed by pemphigus vulgaris (27.3%), dermatitis herpetiformis (15.3%), Darier's disease (14.7%), pemphigus foliaceus (4.7%), epidermolysis bullosa (2%), linear immunoglobulin A dermatosis (2%), paraneoplastic pemphigus (0.7%), and drug reactions (0.7%). DIF studies were applied on 60 cases, out of which complement protein C3c was the most commonly deposited protein (53.3%). Conclusion Our study emphasized the diagnostic role of skin punch biopsy in the proper evaluation of vesiculobullous skin lesions. Histopathology is the cornerstone diagnostic tool in this regard, with DIF being a useful adjunct.

Prognostic significance of epidermal growth factor receptor (EGFR) over expression in urothelial carcinoma of urinary bladder.

BACKGROUND: Epidermal growth factor receptor (EGFR) has been shown to have abnormal expression in many human cancers and is considered as a marker of poor prognosis. Frequency of over expression in bladder cancer has not been studied in our population; therefore we aimed to evaluate the frequency and prognostic significance of EGFR immunohistochemical expression in locoregional population. METHODS: We performed EGFR immunohistochemistry on 126 cases of bladder cancer and association of EGFR expression with tumor grade, lamina propria invasion, deep muscle invasion and recurrence of disease was evaluated. RESULTS: High EGFR expression was noted in 26.2% (33 cases), 15.1% (19 cases) and 58.7% (74 cases) revealed low and no EGFR expression respectively. Significant association of EGFR expression was noted with tumor grade, lamina propria invasion, deep muscle invasion and recurrence status while no significant association was seen with age, gender and overall survival. Kaplan- Meier curves revealed significant association of EGFR expression with recurrence while no significant association was seen with overall survival. CONCLUSION: Significant association of EGFR overexpression with tumor grade, muscularis propria invasion and recurrence signifies its prognostic value; therefore EGFR can be used as a prognostic biomarker in Urothelial bladder carcinoma.

Molecular subtypes of breast cancer in South Asian population by immunohistochemical profile and Her2neu gene amplification by FISH technique: association with other clinicopathologic parameters.

Molecular breast cancer subtypes were defined by gene expression prolife; however, immunohistochemical (IHC) expression can categorize breast cancers analogous to gene expression profiling. We aimed to evaluate distribution of these molecular breast cancer subtypes in our population and their association with clinocopathologic parameters. We retrospectively analyzed 1,104 cases of primary breast cancers over 3 years duration. ER, PR, Her2neu IHC staining, and subsequent fluorescent in situ hybridization studies (Her2neu gene amplification in cases with 2+ IHC staining) were performed to categorize breast cancer subtypes. Luminal A breast cancers were most frequent (45.8%) followed by triple negative (18.6%), luminal B (17.8%) and Her2neu (17.8%) subtypes. We found a strong association of breast cancer subtypes with tumor grade and Ki67 proliferation index with triple negative cancers being associated with higher grade and proliferation index. Significant association was seen with age groups, luminal A subtype occurring at a slightly older age, whereas triple negative and Her2neu cancers were more frequent in younger age group. We found a higher proportion of triple negative cancers in our set up, and they were found to have high-tumor grade and proliferation index along with presentation at younger age. As these cancers are associated with BRCA 1 mutations and abnormal BRCA 1 pathways, we suggest that large scale studies should be done to evaluate BRCA 1 mutations and abnormal BRCA 1 pathways in our population to establish risk factors for this highly aggressive tumor subtype.

Clinicopathologic features of triple negative breast cancers: an experience from Pakistan.

BACKGROUND: Young age breast cancers are quite prevalent in our setup, a significant number of which exhibit triple negative phenotype. These cancers behave in an aggressive fashion and unresponsive to targeted adjuvant therapy. We aimed to evaluate clinical and histopathologic features of triple negative cancers in our population. METHODS: We retrospectively evaluated 1104 cases of primary breast cancers. Immunohistochemical studies for ER, PR and Her2neu followed by Her2neu gene amplification by FISH testing were done to identify 205 (18.6%) cases of triple negative breast cancers. RESULTS: Mean age for triple negative breast cancer patients was 48.4 years (±12.3) and 60% of patients were diagnosed at less than 50 years of age. Although ductal carcinoma was the most frequent histologic type, a meaningful number of cases exhibited metaplastic and medullary like features (10.7% and 5.9% respectively). Similarly geographic necrosis involving more than 40% of tumor and extensive lymphocytic infiltration was a considerable finding. Mean Ki67 index was 45.2% (±25.2) and as a reflection of tumor grade, a significantly higher proportion of cases (66.3%) were under high risk Ki67 category (>30%). CONCLUSION: Triple negative breast cancers typify high grade breast cancers with a higher frequency of atypical medullary and metaplastic histologies. Their prevailing occurrence at a younger age raises question of under lying BRCA mutations in our population. Therefore, we suggest that risk factors including BRCA 1 mutations should be uncovered in reproductive age group breast cancers especially those disclosing basal like phenotype. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/9042440621102239.

Breast lesions in adolescents and young women in Pakistan--a 5 year study of significance of early recognition.

BACKGROUND: Breast cancer is the most frequent malignant disease amongst young women. If we review local data in Pakistan then breast cancer represents approximately a third of all cancers in females. The age standardized incidence rate (ASR) world per 100,000 is 53.8 and crude incidence rate is 30.9. We have observed during our surgical pathology practice and it is also reported by other Asian studies that breast carcinoma is amongst the leading malignancies in the region and the patients are at least a decade younger than counterparts in developed nations. Age is an important issue in effective screening, diagnosis and management of breast cancer, especially in this geographical region where late presentation and poor prognosis are a hallmark of the disease. OBJECTIVE: The aim of this study is to determine the frequency of malignant breast lesions in symptomatic young females presenting with breast lumps. MATERIALS AND METHODS: This is a retrospective study conducted at the Pathology Department, Liaquat National Hospital and Medical College, Karachi. Descriptive and pathology data of malignant breast tumors 1st January 2004 to 31st December 2009 were reviewed, using the departmental archived data. It included both male and female patients up to the age of 25 years. RESULTS: A total of 714 surgical specimens from/of symptomatic breast lesions were received at the pathology department of Liaquat National Medical, in the five years study period, in young females. There were 575 (80%) benign, 119 (16%) inflammatory and 20 (2.8%) malignant lumps. CONCLUSIONS: The obtained data for females only up to 25 years of age suggest a massive burden which requires urgent attention. Early assessment of lesions is essential in order to avoid mortality from malignancies.

Accuracy of frozen section analysis of sentinel lymph nodes for the detection of Asian breast cancer micrometastasis - experience from Pakistan.

BACKGROUND: Intraoperative sentinel lymph node biopsy has now become the standard of care for patients with clinically node negative breast cancer for diagnosis and also in order to determine the need for immediate axillary clearance. Several large scale studies confirmed the diagnostic reliability of this method. However, micrometastases are frequently missed on frozen sections. Recent studies showed that both disease free interval and overall survival are significantly affected by the presence of micrometastatic disease. The aim of this study was to determine the sensitivity and specificity of intraoperative frozen section analysis of sentinel lymph nodes (SLNs) for the detection of breast cancer micrometastasis and to evaluate the status of non-sentinel lymph nodes (non-SLNs) in those patients subjected to further axillary sampling. MATERIALS AND METHODS: We performed a retrospective study on 154 patients who underwent SLN biopsy from January 2008 till October 2011. The SLNs were sectioned at 2 mm intervals and submitted entirely for frozen sections. Three levels of each section submitted are examined and the results were compared with further levels on paraffin sections. RESULTS: Overall 40% of patients (62/154) were found to be SLN positive on final (paraffin section) histology, out of which 44 demonstrated macrometastases (>2mm) and 18 micrometastases (<2mm). The overall sensitivity and specificity of frozen section analysis of SLN for the detection of macrometastasis was found to be 100% while those for micrometastasis were 33.3% and 100%, respectively. Moreover 20% of patients who had micrometastases in SLN had positive non-SLNs on final histology. CONCLUSIONS: Frozen section analysis of SLNs lacks sufficient accuracy to rule out micrometastasis by current protocols. Therefore these need to be revised in order to pick up micrometastasis which appears to have clinical significance. We suggest that this can be achieved by examining more step sections of blocks.

Spectrum of intra-thoracic lesion detected by computed tomography guided fine needle aspiration biopsy.

BACKGROUND: Fine needle aspiration biopsy (FNAB) is a rapid, sensitive and inexpensive procedure for diagnosing benign and malignant palpable lesions. For lesions that are not palpable or deep seated, FNAB can be performed under the guidance of radiological imaging. Our basic objective was to evaluate the spectrum of intrathoracic lesions by using Computed Tomography guided fine needle aspiration biopsy and evaluate its diagnostic yield. METHODOLOGY: It was a retrospective study carried out in the Department of Histopathology, Liaquat National Hospital and Medical College, during the months of August 2011 and August 2012. All patients with pulmonary, mediastinal or paravertebral mass who underwent CT guided intrathoracic biopsy were included in this study. Fine needle aspiration biopsies were performed in the Radiology Department and specimen retrieved was sent in 10% buffered Formalin to the Histopathology Department. All the data was entered and analyzed through SPSS 19.0. RESULTS: A total of 130 cases were evaluated, out of which 108 (83.1%) were pulmonary, 16 (12.3%) were mediastinal and 6 (4.6%) were paravertebral. Conclusive biopsies were possible in 113 cases, while 17 biopsies were inconclusive. In those that showed a conclusive diagnosis, 83.1% were neoplastic and 16.9% were non neoplastic. Of the neoplastic cases, 27 (20.8%) were adenocarcinomas, followed by squamous cell carcinomas (15.4%) and large cell carcinoma, not otherwise specified, (12.3%). CONCLUSION: CT guided fine needle aspiration biopsy is a reliable tool for examination of intrathoracic lesions, with a high rate of conclusive diagnosis.