Background
This 32 year old female presented with blood and protein in her urine. Her left kidney was small on ultrasound with scarring at the upper pole. She also had a recent history of pulmonary embolisms and a radiological diagnosis of focal nodular hyperplasia of the liver. Her glomerulonephritis screen was negative, and the clinical team were considering IgA nephropathy. A renal biopsy of her right kidney was undertaken.

Results/Findings
The biopsy showed glomerulomegaly, ischaemic changes, and one glomerulus contained a hyalinotic scar. Mild patchy interstitial fibrosis and tubular atrophy was present, and there was also mild fibrointimal thickening of interlobular sized arteries. Immunofluorescence was negative. The features raised the possibility of focal segmental glomerulosclerosis (FSGS). Review of the laboratory information system showed a placenta from a previous miscarriage.

The limited renal biopsy findings were not sufficient to explain the level of blood and protein in the urine, but the history of pulmonary embolisms and miscarriage was suggestive of antiphospholipid syndrome (APS). Warfarin anticoagulation prevented routine assessment for the lupus anticoagulant, but a Taipan Snake Venom Time (TSVT) test showed a result at the upper limit of the reference range. Anti-cardiolipin and anti-beta-2 glycoprotein-1 antibodies were negative. The ultrasound appearance of a small left kidney could be indicative of renal artery stenosis, which is associated with APS.

Conclusion
The classic renal involvement in APS is thrombotic microangiopathy, which was not present in this biopsy, but the microscopic features did not explain the clinical presentation. This case highlights the role of pathologists in bringing together the clinico-pathological correlation. We raised the possibility of APS to the renal physicians given the clinical history as a potential unifying diagnosis, allowing for appropriate investigation. Sero-negative APS or another coagulopathy remains a possibility.
 

Background:
Sertoli leydig cell tumors are rare tumors(<0.5% of ovarian neoplasms). Most of the tumors present in a young age. Patients present with virilization or isosexual precocity. Tumors can be solid (fleshy pale yellow) or cystic with a mean size of 12-14 cm. These tumors usually affect young adults and are seen in one or both testicles with microcalcifications. Sertoli leydig cell tumors express FOXL2 in 50% of cases. Mutations in DICER1 are present in 60% of cases.

Purpose: To evaluate the NKX3.1 expression by immunohistochemistry in normal testicular parenchyma, in Sertoli cell tumors and Sertoli Leydig cell tumors of testes and ovary.

Methods: Retrospective observational study.
Place and Duration of Study: Shaukat Khanum Memorial Cancer Hospital and Research Centre, Lahore 2011-2021.
Methodology: We used immunohistochemistry to evaluate the positivity and loss of nuclear expression of NKX3.1 in sertoli cell tumor (11 cases), sertoli Leydig cell tumor (31 cases) and in normal testicular parenchyma (7 cases).

Results: 10 out of 10 cases of benign testicular parenchyma expressed positivity with nuclear staining of NKX3.1 in Sertoli cells. 2 out of 11 Sertoli cell tumors expressed positivity with nuclear positivity of NKX3.1 in Sertoli cell component (18.18%) and 9 of the cases showed loss of staining of NKX3.1 (81.8%). All Sertoli Leydig cell tumors showed loss of staining of NKX3.1.

Conclusion: Nuclear expression of NKX3.1 is seen in Sertoli cells of normal testicular parenchyma. This staining is lost in Sertoli cell tumors and Sertoli Leydig cell tumors.
 

Background:
Clear cell renal cell carcinoma generally occurs in adults and comprises 70% of all epithelial renal tumours. BAP1 (BRCA1 associated protein 1 on chromosome 3) is a commonly mutated gene in clear cell renal cell carcinoma. Aim of the study was to evaluate the prognostic significance of BAP1 by immunohistochemistry in clear cell renal cell carcinoma.

Methods:
It was a descriptive case series in which data was retrospectively collected. In this study, there were 142 cases of which 110 cases were of clear cell renal cell carcinoma, 16 cases were papillary renal cell carcinoma and 16 cases were of chromophobe renal cell carcinoma. Immunohistochemistry was used to evaluate the loss of nuclear expression of BAP1.

Results:
Loss of BAP1 was observed in 60% of cases of clear cell renal cell carcinoma. 27% of grade 1 tumours, 62% of grade 2 tumours, 65% of grade 3 tumours and 66% of grade 4 tumours showed loss of BAP1. Loss of BAP1 was observed in 54% cases of stage 1 tumours, 72% of stage 2 tumours and 66% of stage 3 tumours. Our study showed loss of BAP1 in 67% of cases with tumour necrosis, in 75% of cases with sarcomatoid features and in 60% of patients with distant metastasis.

Conclusions:
We conclude that the loss of BAP1 nuclear expression is associated with poor prognostic features. i.e., higher grade, higher stage, tumour necrosis, sarcomatoid features and distant metastasis leading to death of patients. These finding lend support to the idea of doing BAP1 immunohistochemistry in routine labs for determining prognosis.
 

Background: The poor prognostics of central nervous system tumours are a clinical problem that is in part due to the tight substance control from the blood-brain barrier. Microbubbles combined with focused ultrasound is a developing method to overcome this issue with a strong evidence base for effectiveness and safety, a topic that will be discussed.
Purpose: This review will cover a collection of pre-clinical and clinical trials from the past twelve years, looking at the uses of different microbubbles. Trials using different bubbles, including Definity, Optison, and Sonovue, will be discussed with uses in sonobiopsy/liquid biopsy, chemotherapy drug delivery and immunotherapy. It will also look at current discussions around the safety and tissue effects of microbubble use on the blood-brain barrier, covering any changes noticed in histology and radiology or adverse effects experienced by patients in clinical trials.
Method: A search was conducted on Web of Science to locate clinical trials using microbubbles in central nervous system tumour diagnosis and treatment.
Results: The search located sixteen clinical trials on the uses of microbubbles in central nervous system tumours, covering drug delivery, immunotherapy and liquid biopsy. This includes clinical and pre-clinical trials with a focus on the implementation of microbubbles and assessing the safety of opening the blood-brain barrier.
Conclusion: Central nervous system tumours are a big area of development with microbubbles and focused ultrasound regimes being developed for diagnosis and treatment methods. It’s a promising area with published early clinical trials and many ongoing or proposed clinical trials happening to explore clinical implementation.
 

Background: Oral potentially malignant lesions (OPML) manifest as white or red lesions, exhibiting histological alterations. While many OPMLs revert to normal epithelium, 9-33% may progress to oral cancer. Over the past decade, UK oral cancer cases surged by 58%, with a continued upward trend projected. In 2020, there were 200,000 OPML referrals, resulting in 8,722 oral cancer diagnoses.

Purpose: The key molecular events leading to OPML and progression to oral cancer are poorly understood. Much of the published research uses cells cultured in monolayer which does not physiologically represent the in vivo situation. We aimed to validate tissue-engineered mucosal constructs as a tool for defining key molecular events involved in cancer progression.
Methods: Normal, dysplastic, and OSCC keratinocytes were used to generate full-thickness tissue-engineered models, validated against native tissue. Next-generation RNA sequencing and DNA methylation profiling identified differentially expressed and methylated genes. Bioinformatics analysis evaluated gene set pathways and identified genes of interest. Validation was conducted using Sanger sequencing and qPCR, with the hypomethylating agent decitabine assessed for potential gene function restoration.

Results: The tissue-engineered constructs closely resembled native tissue, with dysplastic models exhibiting disorganized epithelial architecture and cellular atypia. RNA and DNA methylation sequencing revealed 1070 downregulated and 1907 upregulated genes, and 1209 hypermethylated and 1791 hypomethylated promoter regions in dysplastic models compared to normal. Integration analysis identified 59 genes downregulated and hypermethylated, associated with key pathways including choline metabolism, p53 signaling, antigen processing, and cytokine signaling. Decitabine effectively restored the function of specific tumor suppressor genes in the dysplastic model.

Conclusion: The study demonstrates the reliability of tissue-engineered models in reproducing cancer progression in vitro. Dysplastic models showed significant molecular alterations associated with oral cancer progression. These findings may offer insights for developing therapeutic targets to improve outcomes for early-stage OSCC patients.
 

Background: Artificial Intelligence (AI) has been shown to help in the identification and quantification of digital tissue biomarkers in several cancers. However, there has been little exploration of its role in head and neck squamous cell carcinoma (HNSCC).
Purpose: To determine whether morphometric analysis using a custom-trained classifier is useful in the objective assessment of clinicopathological, molecular, mutational, and survival correlation in HNSCC.
Methods: The retrospective study used whole slide images (WSI) of HNSCC from the publicly available datasets (The Cancer Genome Atlas and The Cancer Imaging Archive) and an AI-based open-source software (QuPath) for image analysis. In the morphometric study, epithelial, stromal, and immune cells in WSI of normal oral mucosa (NOM), dysplasia (OED), and HNSCC were annotated, followed by morphological features extraction and comparison using ANOVA and Student’s t-test. For classifier development, two classifiers, artificial neural network-multilayer perceptron (ANN-MLP) and Random trees (Rtrees) were trained with epithelium, stroma, immune cells, and mitotic figures in WSI of NOM, OED, and HNSCC WSIs. The performance of the classifiers was tested on unseen HNSCC WSIs for automatic segmentation followed by downstream analysis.
Results: Student t-test and ANOVA revealed statistically significant differences between most morphological features of epithelial, stromal, and immune cells between NOM, OED, and HNSCC (p<0.05). The ANN-MLP classifier performed better than Rtrees for the automatic segmentation of epithelial, stromal, and immune cells, with F1 scores of 0.78, 0.79, and 0.82 respectively. The downstream analysis showed a significant correlation between morphometric features and prognosis in HNSCC (p<0.05). The F1 score of the ANN-MLP classifier for atypical mitosis was 0.79 and both the automatic and manual counts were significantly correlated with prognosis in HNSCC (p<0.05).
Conclusion: Morphometric assessment using a custom-trained classifier can potentially serve as a tissue biomarker for objective differentiation between dysplasia and cancer, prognosis determination, and risk stratification.