Specimen Adequacy and Clinicopathological Evaluation of Inflammatory Bowel Disease Colorectal Biopsies in Cipto Mangunkusumo Hospital Jakarta

Background: Colorectal mucosal biopsies account for majority of daily practice specimens in the field of gastrointestinal pathology. Most of them were sent for inflammatory bowel disease (IBD) evaluation. Due to broad spectrum of histologic findings and different stage of disease, pathologists often find difficulties on calling out diagnosis of IBD. This study aims to evaluate the adequacy of specimen, clinical characteristics, endoscopy and histologic findings of colorectal biopsies from IBD and non-IBD patients.Method: This cross-sectional study included 49 IBD and 49 non-IBD cases registered in the archives of Anatomical Pathology Department, Cipto Mangunkusumo National Referral Hospital Jakarta in 2019. The samples were evaluated for adequacy of specimen, clinical, endoscopy and histologic findings.Results: Most samples were adult with a slight female predominance. All request forms contained the clinical working diagnosis but only 26.5% and 20.4% of them provided clinical history and endoscopy findings. During histology evaluation, 59.2% of the specimen was considered sub-optimal. The most common histologic findings in both IBD and non-IBD groups were diffuse lymphoplasmacytic infiltration in the lamina propria and crypt distortion. Fibrosis/collagen deposition was found in 38.8% of IBD population compared to 12.2% of non-IBD population (p=0.003). Most IBD cases (55.1%) were categorized as active phase IBD.Conclusion: Since the most common histologic findings in IBD patients were also found in non-IBD patients, IBD diagnosis based on histopathology alone is highly inadvisable. However, there were also lack of clinical data and endoscopy findings in most of colorectal biopsies sent for IBD evaluation. Along with overlapping and unspecific morphology and suboptimal events during histological evaluation, all of this complicated the diagnosis of IBD. Hence multidisciplinary approach is required for a better IBD diagnosis and treatment.

Utility of Colonoscopic Biopsies in Histomorphological Spectrum of Colorectal Lesions - A Study in a Tertiary Care Centre in Aurangabad Region of Maharashtra

BACKGROUND Colonoscopy is considered as a diagnostic procedure of choice for patients presenting with chronic diarrhoea / bloody diarrhoea. Colonoscopic mucosal biopsies have shown to be accurate indicators of extent of the involvement of colon in inflammatory bowel disease. Also, colonoscopy is still considered as the gold standard in cancer surveillance. The present study was undertaken to find out the utility of colonoscopic biopsies in histomorphological spectrum of colorectal lesions. METHODS This was a three-year retrospective study carried out in the Department of Pathology, at MGM Medical College, Aurangabad. Patient’s clinical details, colonoscopic findings and apparent pathology were noted in 206 cases along with final histopathological diagnosis. Biopsies which showed significant findings were included in the study. RESULTS In the study, the male to female ratio was 2.4 : 1. Out of 206 colorectal biopsies, 146 (70.8 %) were non-neoplastic and 60 (29.2 %) were neoplastic lesions. Amongst non-neoplastic lesions, maximum cases were of chronic colitis with 4 cases of granulomatous colitis and 30 cases of inflammatory bowel disease. Adenocarcinoma was the most common histologic type in neoplastic lesions (66.6 %), with rectum being the most common site followed by colonic polyps (25 %). As far as age group was concerned, the 2nd decade was dominated by chronic colitis, 3rd decade by adenomatous polyps and 5th decade by adenocarcinoma. CONCLUSIONS Histomorphological profile of colorectal biopsies has a wide spectrum ranging from infectious diseases, inflammatory bowel disorders to colorectal malignancies. Histopathology with correlation of clinical and endoscopic findings plays a major role in accurate diagnosis of colorectal lesions. KEYWORDS Colonoscopy, Biopsies, Neoplastic, Non-Neoplastic, Colitis, Adenocarcinoma

HISTOPATHOLOGICALANALYSIS OF TUMOURS AND TUMOUR-LIKE LESIONS OF COLON AND RECTUM

Background:Tumours and tumour like lesions of the colon and rectum have overlapping clinical presentation and may be difcult to diagnose on clinical grounds alone. Histopathological examination although helpful to arrive at correct diagnosis, at times may be difcult and may require ancillary tests such as immunohistochemistry. Materials and Methods: A prospective cross sectional study enrolled 125 cases of colorectal biopsies after histopathological conrmation of tumours and tumour like lesions. The specimens were received at the Department of Pathology, DMCH, Laheriasarai, Bihar from December 2019 to November 2020. Results: Out of 125 cases, 32 cases were benign tumours, 36 cases were malignant tumours and 57 cases were tumour like lesions. Most common age range was 51-60 years with mean age of 38 years. Males comprised 76.74% and females 23.26% with ratio of 3:1. Juvenile polyp was the commonest lesion. Among benign tumours, tubular adenoma was the most common (19.2%), adenocarcinoma was most common malignant tumour (25.6%) and juvenile polyp was the most common tumour like lesion (27.2%). Left side was most common side (66.67%) and rectum was the most common site (48.8%). Conclusions: The commonest indications for colorectal biopsies were tumours and tumour like lesions. Juvenile polyp was the most common tumour like lesion, tubular adenoma was the most common benign tumour and adenocarcinoma was the most common malignant tumour. Histopathological examination is a gold standard for conrmation of all tumour and tumours like lesions of colon and rectum.

TOFACITINIB INHIBITS INFLAMMATORY CYTOKINES FROM ULCERATIVE COLITIS AND HEALTHY MUCOSAL EXPLANTS AND IS ASSOCIATED WITH P-STAT1/3 REDUCTION IN T-CELLS

Introduction: Poor translatability of animal disease models has hampered development of new inflammatory bowel disorders (IBD) therapeutics. We describe a preclinical, ex vivo system using freshly obtained and well-characterized human colorectal tissue from ulcerative colitis (UC) and healthy control (HC) participants to test potential therapeutics for efficacy and target engagement, using the JAK/STAT inhibitor Tofacitinib (TOFA) as a model therapeutic. Methods: Colorectal biopsies from HC and UC were cultured and stimulated with multiple mitogens +/-TOFA. Soluble biomarkers were detected using a 29-analyte multiplex ELISA. Target engagement in CD3+CD4+ and CD3+CD8+ T cells was determined by flow cytometry in PBMC and isolated mucosal mononuclear cells (MMC) following activation of STAT1/3 phosphorylation. Data were analyzed using linear mixed effect modeling, t-test, and analysis of variance. Biomarker selection was performed using penalized and Bayesian logistic regression modeling, with results visualized using uniform manifold approximation and projection (UMAP). Results: Under baseline conditions, 27/29 biomarkers from UC were increased versus HC. Explant stimulation increased biomarker release magnitude, expanding the dynamic range for efficacy and target engagement studies. Logistic regression analyses identified the most representative UC baseline and stimulated biomarkers. TOFA inhibited biomarkers dependent on JAK/STAT signaling. STAT1/3 phosphorylation in T-cells revealed compartmental differences between PBMC and MMC. Conclusions: Immunogen stimulation increases biomarker release in similar patterns for HC and UC, while enhancing the dynamic range for therapeutic efficacy studies. This work demonstrates the power of ex vivo human colorectal tissue as preclinical tools for evaluating target engagement and downstream effects of new IBD therapeutic agents.

Kayexalate-induced colitis and rectal stricture

A 57-year-old man underwent emergency laparoscopic loop colostomy for acute recto-sigmoid obstruction. He was hospitalised 2 months previously, at another facility for diabetic ketoacidosis (DKA) and hyperkalaemia. He had no gastrointestinal symptoms prior to the hospitalisation. Both surgical exploration and intraoperative sigmoidoscopy showed ulcerations of sigmoid colon and proximal rectum with a pinhole stricture in mid-rectum. After ruling out all aetiologies, and due to persistence of the colonic ulcerations on a follow-up colonoscopy, a diagnosis of Crohn’s colitis was made, and the patient was started on infliximab and 6-mercaptopurine (6-MP). Six months later, on rereview of all the biopsies, it was noted that a key element of presence of crystals suggestive of Kayexalate on the initial colorectal biopsies was missed. It was later found out that the patient had received rectal Kayexalate for treatment of DKA at the other facility. Hence, infliximab and 6-MP were both discontinued. All the colonoscopies, following the discontinuation of the medications, showed complete resolution of colitis but persistence of the mid-rectum stricture. This was treated with a fully covered metal stent for 12 weeks with only partial improvement of the stricture. He was hence referred for ultra-low anterior resection of rectum and take down of colostomy.

Glycan array analysis of Pholiota squarrosa lectin and other fucose-oriented lectins

The α(1,6)fucose residue attached to the N-glycoprotein core is suspected to play an essential role in the progression of several types of cancer. Lectins remain the first choice for probing glycan modifications, although they may lack specificity. Thus, efforts have been made to identify new lectins with a narrower core fucose (CF) detection profile. Here, we present a comparison of the classical Aleuria aurantia lectin (AAL), Lens culinaris agglutinin (LCA) and Aspergillus oryzae lectin (AOL) with the newer Pholiota squarrosa lectin (PhoSL), which has been described as being specific for core fucosylated N-glycans. To this end, we studied the binding profiles of the four lectins using mammalian glycan arrays from the Consortium of Functional Glycomics. To validate their glycan specificity, we probed AOL, LCA and PhoSL in western-blot assays using protein extracts from eight common colorectal cancer (CRC) lines and colorectal biopsies from a small cohort of patients with CRC. The results showed that (i) LCA and PhoSL were the most specific lectins for detecting the presence of CF in a concentration-dependent manner; (ii) PhoSL exhibited the highest N-glycan sequence restriction, with preferential binding to core fucosylated paucimannosidic-type N-glycans, (iii) the recognition ability of PhoSL was highly influenced by the presence of terminal N-acetyl-lactosamine; (iv) LCA bound to paucimannosidic, bi-antennary and tri-antennary core fucosylated N-glycans and (v) AOL and AAL exhibited broader specificity towards fucosylation. Together, our results support the choice of LCA as the most appropriate lectin for CF detection, as validated in protein extracts from CRC cell lines and tissue specimens from patients with CRC.

Histopathological analysis of tumours and tumour-like lesions of colon and rectum

Background: Tumours and tumour like lesions of the colon and rectum have overlapping clinical presentation and may be difficult to diagnose on clinical grounds alone. Histopathological examination although helpful to arrive at correct diagnosis, at times may be difficult and may require ancillary tests such as immunohistochemistry. Materials and Methods: A prospective cross sectional study enrolled 125 cases of colorectal biopsies after histopathological confirmation of tumours and tumour like lesions. The specimens were received at the Department of Pathology, of TUTH, from December 1, 2015 to November 30, 2016. Results: Out of 125 cases, 32 cases were benign tumours, 36 cases were malignant tumours and 57 cases were tumour like lesions. Most common age range was 51-60 years with mean age of 38 years. Males comprised 76.74% and females 23.26% with ratio of 3:1. Juvenile polyp was the commonest lesion. Among benign tumours, tubular adenoma was the most common (19.2%), adenocarcinoma was most common malignant tumour (25.6%) and juvenile polyp was the most common tumour like lesion (27.2%). Left side was most common side (66.67%) and rectum was the most common site (48.8%). Conclusions: The commonest indications for colorectal biopsies were tumours and tumour like lesions. Juvenile polyp was the most common tumour like lesion, tubular adenoma was the most common benign tumour and adenocarcinoma was the most common malignant tumour. Histopathological examination is a gold standard for confirmation of all tumour and tumours like lesions of colon and rectum.

How many serial sections are needed to detect apoptosis in endoscopic biopsies with gastrointestinal graft versus host disease?

AimsThe hallmark histological feature of acute gastrointestinal graft versus host disease (GI GVHD) is epithelial apoptosis. This is the first formal evaluation of how many serial sections are required to consistently detect apoptotic bodies in endoscopic biopsies from various GI locations in patients with clinically validated GI GVHD.Methods, results and conclusionsAssessment of 1008 serial sections showed that apoptotic bodies are uniformly distributed among such sections of gastric, duodenal and colorectal biopsies from these patients. Assessment of 59 further biopsies showed that assessing 12 serial sections should suffice to detect GVHD in gastric, duodenal and colorectal biopsies using thresholds of one apoptotic body per biopsy fragment or one apoptotic body per 4 mm2. Assessing 12 serial sections should also suffice to detect GVHD in duodenal and colorectal biopsies using the threshold of 6 apoptotic bodies per 10 contiguous crypts, but it remains uncertain whether this assessment and threshold can be applied to gastric biopsies.