Reliable Cerebrospinal Fluid Cytology and Immunocytochemistry Reporting over an Extended Period by the Addition of Aldehyde and Osmolyte-Based Preservative Solution.

INTRODUCTION: The cytological examination of cerebrospinal fluid (CSF) is an important investigation in the workup of various inflammatory, malignant, or traumatic disorders of the central nervous system. The cells in the CSF lyse and degenerate at a very fast rate owing to its low tonicity, buffering capacity, redox potential, and pH, making it crucial to examine it within 2 h of sampling. We have hereby designed an aliphatic aldehyde, osmolyte, metal halide, and a buffer-based solution which will preserve the cellular components of CSF for 48 h. METHODS: Thirty-nine CSF specimens were examined within 2 h of collection, and this reading was recorded as time zero reading. The CSF specimens were then divided into two tubes with (i) pre-servative:CSF ratio of 1:5; and (ii) no preservative. Total and differential leukocyte counts and immunocytochemistry were performed on the paired specimens at 24 h and 48 h and were compared with the readings at zero hours. RESULTS: The preservative-containing CSF showed significantly higher cellularity than the undiluted samples at 24 h and 48 h (p < 0.001). Median cell counts observed in the preservative-containing CSF were 5 times and 12 times higher than in the undiluted CSF. Neutrophils, lymphocytes, and RBCs showed immunopositivity for MPO, CD45, and GLUT-1 at both time intervals. CONCLUSION: Adding the prepared preservative solution to CSF in the ratio of 1:5 can optimally preserve the CSF cells for absolute and differential quantitation, morphological assessment, and immunological testing at a later date.

Vulvar dermatoses-Can a pattern-based approach improve diagnostic yield?

BACKGROUND: Vulvar dermatoses (VD) pose a formidable challenge to clinicians and pathologists owing to various factors. The factors included are the histopathological heterogeneity of the vulva, moist and frictional environment, and the limited experience of gynecologists and general histopathologists in this field. To address this issue, the International Society for the Study of Vulvovaginal Disease (ISSVD) proposed a histopathological tissue reaction-based classification system for VD. Therefore, we attempted to study the utility of the 2006 ISSVD classification in reporting VD. We further evaluated if a dermatopathologist review could improve the diagnostic yield. MATERIALS AND METHODS: The vulvar biopsy reports (N = 106) were retrieved from histopathology case files, out of which benign non-infectious conditions (n = 55) were included in the study. The diagnosis retrieved from the case files was considered the initial diagnosis. Three dermatopathologists reviewed each biopsy, and a tissue reaction pattern/diagnosis was assigned as per ISSVD 2006, and this was considered a review diagnosis. The initial and review diagnoses were compared and analyzed. We further studied and analyzed the effect of the dermatopathologist's review on the diagnostic yield. RESULTS: The sclerotic pattern (34.6%) was the commonest tissue reaction pattern, followed by spongiotic (18%) and acanthotic patterns (14.5%) independently or in combination. The non-specific/descriptive report rate was significantly decreased following 2006 ISSVD and the dermatopathologist's review (83.6% vs.1.8%). CONCLUSION: Rendering tissue reaction patterns to vulvar biopsies will enable a comprehensive understanding of lesions and aid in clinically relevant reporting. In addition, dermatopathologists' review of difficult vulvar biopsies increases the diagnostic yield.

Clinico-Etiologic Profile of Macroscopic Hematuria in Children: A Single Center Experience.

OBJECTIVE: To study the demographic, clinical and etiological profile of macroscopic hematuria in children presenting to a tertiary care hospital. METHODS: This prospective observational study, conducted between January, 2018 and December, 2019, enrolled children aged 3 months to 12 years, presenting with gross hematuria. RESULTS: Of the 62 children (44 males) enrolled, (mean (SD) age of 7.3 (2.6) years), glomerular hematuria was seen in 59.7%. Post-infectious glomerulonephritis was the commonest etiology of glomerular hematuria; hypercalciuria and renal calculi predominated among non-glomerular hematuria. After a median (IQR) follow up of 8 (6,14.2) months, microscopic hematuria persisted in 10 (7, glomerular hematuria) children. The median time to resolution of gross as well as microscopic hematuria tended to be longer in glomerular etiologies. CONCLUSION: Majority of children with gross hematuria had glomerular etiologies, thus requiring monitoring and follow-up.

Best practices of handling, processing, and interpretation of small intestinal biopsies for the diagnosis and management of celiac disease: A joint consensus of Indian association of pathologists and microbiologists and Indian society of gastroenterology.

The Indian Association of Pathologists and Microbiologists (IAPM) and Indian Society of Gastroenterology (ISG) decided to make a joint consensus recommendation for handling, processing, and interpretation of SI biopsies for the diagnosis and management of celiac disease (CD) recognizing the inhomogeneous practice of biopsy sampling, orientation, processing, and interpretation. A modified Delphi process was used to develop this consensus document containing a total of 42 statements and recommendations, which were generated by sharing the document draft, incorporating expert's opinion, followed by three cycles of electronic voting as well as a full-day face-to-face virtual ZOOM meeting and review of supporting literature. Of the 42 statements, 7 statements are on small intestinal (SI) biopsy in suspected patients of CD, site and the number of biopsies; 7 on handling, fixative, orientation, processing, and sectioning in pathology laboratories; 2 on histological orientation; 13 statements on histological interpretation and histological grading; 3 on the assessment of follow-up biopsies; 2 statements on gluten-free diet (GFD)-nonresponsive CD; 4 on challenges in the diagnosis of CD; 2 statements each on pathology reporting protocol and training and infrastructure in this area. The goal of this guideline document is to formulate a uniform protocol agreed upon both by the experienced pathologists and gastroenterologists to standardize the practice, improve the yield of small bowel biopsy interpretation, patients' compliance, overall management in CD, and generate unified data for patient care and research in the related field.

Histological analyses of trucut liver biopsies from patients with noncirrhotic portal fibrosis and extra-hepatic portal vein obstruction.

BACKGROUND: Both noncirrhotic portal fibrosis (NCPF) and extrahepatic portal venous obstruction (EHPVO) are important causes of noncirrhotic portal hypertension (PH) in the Asian region. In this study, we analyzed the histopathological changes of liver needle-core biopsies from patients with NCPF and EHPVO. PATIENTS AND METHODS: The patients were diagnosed as per the Asia Pacific Association for the Study of Liver (APASL) criteria. Minimum adequacy criteria for liver core biopsies were defined, and finally, 69 liver biopsies from patients with NCPF and 100 liver biopsies from patients with EHPVO were analyzed. All histological parameters were predefined, and three experienced pathologists analyzed the biopsies after reaching consensus. Institute ethics committee clearance was taken. RESULTS: Although some histological features were overlapping, phlebosclerosis of intra-hepatic branches of the portal vein (PV), periportal aberrant vascular channels, remnant portal tracts, and hepatic fibrosis beyond the portal tracts without the formation of complete hepatic nodules (P < 0.001 for all) were common histological characteristics of NCPF on core-needle liver biopsies; while maintained lobular architecture, nonspecific dilatation of PV branches, absence of intra-hepatic PV phlebosclerosis, aberrant vascular channels, and significant fibrosis were characteristics of EHPVO. CONCLUSIONS: Despite the considerable histological overlap between NCPF and EHPVO, careful histological evaluation, supplemented by clinical features, radiological and biochemical findings can help in making a conclusive diagnosis. Patients with NCPF and EHPVO with clinical jaundice show transaminitis, high serum alkaline phosphatase level, more variceal bleed, and histological evidences of nodular regenerative hyperplasia.

Differentiating Biliary Atresia From Idiopathic Neonatal Hepatitis: A Novel Keratin 7 Based Mathematical Approach on Liver Biopsies.

BACKGROUND AND AIMS: Differentiating biliary atresia (BA) from idiopathic neonatal hepatitis (INH) is vital in routine pediatric practice. However, on liver biopsy, few cases offer a diagnostic challenge to discriminate these entities with certainty. Bile ductular reaction (DR), intermediate hepatobiliary cells (IHBC) and extra-portal ductules (EPD) indicate progenitor cell activation, as a response to various hepatic insults. The present study aims to quantify DR, IHBC and EPD by Keratin 7 (CK7) immunohistochemistry (IHC) in BA and INH and to devise a mathematical approach to better differentiate the two, especially in histologically equivocal cases. METHODS: A total of 98 cases were categorized on biopsy as BA, INH or equivocal histology, favoring BA or INH. CK7 DR mean, IHBC mean and EPD mean values were compared between BA and INH. A formula was derived to help distinguish these two entities, the cut-off value, sensitivity and specificity of which were determined by receiver operating characteristic (ROC) curve. This formula was applied and validated on histologically equivocal cases. RESULTS: Univariate logistic regression revealed significant difference between BA and INH with respect to CK7 DR and CK7 EPD mean (p < 0.001 in both); however, CK7 IHBC mean was not significant (p = 0.08). On multivariate logistic regression, only CK7 DR had significant impact on diagnosis (p < 0.001). A formula: (CK7 DR)2 + (CK7 EPD)/(CK7 IHBC) was derived to help distinguish BA from INH. Cut off value of 10.5 and above, determined by ROC curve, favored a diagnosis of BA (sensitivity= 93.4%, specificity= 94.6%). Histologically equivocal and discrepant cases could be correctly categorized using this formula. CONCLUSIONS: Formula using CK7 IHC parameters may aid pathologists better distinguish BA from INH, especially in histologically equivocal cases.

Myofibromatosis.

INTRODUCTION: Myofibromatosis is a distinctive mesenchymal disorder occurring predominantly in childhood, which on microscopy shows peripheral light areas of spindle cells and central cellular areas of primitive oval to spindle cells arranged around hemagiopercytomatous vessels. PDFGRB mutations in the familial and multifocal sporadic forms and SRF-RELA fusions in the cellular variants have been identified. The index case is being presented to discuss the clinico-pathological features, differential diagnosis, and management of the lesion. CASE PRESENTATION: An 11-year-old male presented with an infraorbital mass of 3 months duration. The mass was excised and microscopy revealed the morphological features of myofibroma with tram-track SMA immunopositivity. Nodular fasciitis and fibromatosis were the differentials considered. CONCLUSION: The SRF-RELA gene fusion may represent a subset that in the future may be used to differentiate these myofibromas/myopericytomas from the ACTB-GLI fusion myopericytomas, and PDGFRB may be used to perhaps separate out familial myofibromas from other myofibromas.

Pediatric fine-needle aspiration cytology: An audit of 266 cases of pediatric tumors with cytologic-histologic correlation.

OBJECTIVES: Fine-needle aspiration cytology (FNAC), a well-accepted minimally invasive diagnostic technique utilized in adults, is gradually gaining ground for pediatric patients as well. However, there are very few comprehensive reports in the literature on utility of FNA in pre-operative diagnosis of pediatric tumors. MATERIAL AND METHODS: An observational study was conducted at a cancer research center and a pediatric tertiary care hospital over a 5-year period. A cytologic-histologic correlation was performed for FNACs performed in pediatric patients for a clinical diagnosis of neoplastic lesions at both the centers. Relevant clinical details and histopathology, wherever available, were retrieved. Sensitivity, specificity, and accuracy of FNAC in diagnosis of malignant lesion were calculated from the cases with available histologic correlation. RESULTS: Of the 266 cases included, there was a slight male predominance with lymphadenopathy being the most common presentation and non-Hodgkin's lymphoma as the most frequent diagnosis in cases clinically suspected to have a neoplasm. Histologic correlation was available in 112 cases with 100% concordance in liver and kidney tumors. Few rare cytologic diagnoses such as papillary renal cell carcinoma, mesenchymal hamartoma of the liver, and thymolipoma could be accurately rendered on FNAC smears in conjunction with the clinic-radiologic features. The sensitivity, specificity, and accuracy of FNA in diagnosing malignant pediatric tumors were found to be 100%, 92.6%, and 97.7%, respectively. CONCLUSION: The present study underscores the high sensitivity and accuracy of FNAC in diagnosis of pediatric tumors, both in superficial and deep-seated locations. Awareness of the cytomorphologic features and clinic-radiologic correlation may assist the cytopathologists in rendering a precise diagnosis of rare pediatric tumors as well.

An Unusual Presentation of a Dermoid Cyst Mimicking a Chalazion.

The authors report the case of a 6-year-old boy who presented to their outpatient department with complaints of progressively increasing swelling on the left upper eyelid since birth. The swelling was excised and was found to be firmly attached to the tarsus. Histopatho-logical examination reported a cyst lined by stratified squamous epithelium with presence of adnexal structures in the subepithelium. Therefore, the diagnosis of tarsal dermoid cyst was made. [J Pediatr Ophthalmol Strabismus. 2020;57:e41-e42.].

Reliable cerebrospinal fluid cytology reporting over an extended period: Is it possible by adding formol saline and ethylenediaminetetraacetic acid as cell preservative?

BACKGROUND: Reporting cerebrospinal fluid (CSF) cytology within a narrow time frame is crucial as it is often indicated in critically ill patients and moreover, the cells in CSF are highly labile and tend to decline rapidly on standing. However, due to various logistic issues, delay in reporting is inevitable at times, especially if ancillary tools are required. In this study, we examine the effect of using formol saline and ethylenediaminetetraacetic acid (EDTA) as a preservative on the cellular composition of CSF at 18, 24, and 48 hours of preservation. METHODS: Thirty CSF specimens were examined within 2 hours of collection and this reading was recorded as time zero reading. The CSF specimens were then divided in three tubes with: (a) preservative:CSF ratio of 1:1; (b) preservative:CSF ratio of 1:5; and (c) no preservative. Total and differential leucocyte counts and immunocytochemistry were performed on the three specimens at 18, 24, and 48 hours and were compared with the readings at 0 hour. RESULTS: Preserved CSF (in the ratio of 1:5) showed no significant decrease in the number of cells at 18 hours (P = .4), 24 hours (P = .3), and 48 hours (P = .1). Cellularity decreased by 8.5%, 22%, and 40% at 18, 24, and 48 hours, respectively. Cell morphology and antigenicity were well preserved at all the three time intervals. CONCLUSION: Formol saline and EDTA, when mixed with the CSF in the ratio of 1:5, can preserve significant cellularity for up to 24 hours.

Benign vascular anomalies: A transition from morphological to etiological classification.

The International Society for the Study of Vascular Anomalies (ISSVA) devised a multidisciplinary etiopathogenesis based approach to classify benign vascular anomalies into tumors and malformations. This classification scheme has major therapeutic and prognostic implications as treatment modalities differ for both the categories. Inappropriate usage of the term "hemangioma" for etiopathogenetically distinct entities is commonly seen in clinical practice leading to delivery of incorrect treatment to the patients. We aimed to study the histomorphological and immunohistochemical features of benign vascular anomalies for their precise histopathological classification. A total of 48 cases diagnosed over a period of 3.5 years were reviewed and reclassified into vascular tumors and malformations based on ISSVA classification and prototypical histopathological features. Biopsies were reviewed based on 5 histopathological criteria viz. endothelial morphology, mitotic activity, intralesional nerve bundles, intralesional inflammation, and prominent vessel type. A panel of GLUT-1, WT-1, and Ki-67 was performed in each case. Seven cases of infantile hemangioma, 4 cases each of non-involuting congenital hemangioma and pyogenic granuloma, and 33 cases of vascular malformations were diagnosed. Endothelial cell morphology (p < 0.001), mitotic activity (p < 0.001), and intralesional nerve bundles (p < 0.001) were found to be statistically significant in differentiating hemangioma from malformations. GLUT-1 (p < 0.001) and Ki-67 labeling index (p < 0.001) were useful to distinguish infantile hemangioma from vascular malformations. To conclude, the ISSVA classification of benign vascular anomalies can be reliably done on histopathology. However, every case must be interpreted in the light of clinical and radiological features.

Non-malignant fibroblastic/myofibroblastic tumors in pediatric age group: Clues and pitfalls to the cytological diagnosis.

INTRODUCTION: Fibroblastic/myofibroblastic tumors constitute 12% of all pediatric soft tissue tumors with the majority of them belonging to the benign and intermediate prognostic categories. They are often misdiagnosed owing to their variable clinical presentation and unusual microscopic features. The diagnosis, specially cytological diagnosis of benign and intermediate categories is difficult due to paucity of cellular component and increased amount of extracellular matrix as compared to malignant ones. We hereby discuss the Fine needle aspiration cytology (FNAC) findings of non-malignant fibroblastic/myofibroblastic lesion in the pediatric age group encountered at our institute. METHODS: All the benign and intermediate fibroblastic/myofibroblastic/fibroadipocytic lesions (age 0-12 years) diagnosed on FNAC over a period of 3½ years (Jan 2016- July 2019), with availability of corresponding histopathology were included in the study. RESULTS: A total of seven pediatric benign and intermediate fibroblastic/myofibroblastic lesions with histopathological confirmation were identified which included Infantile digital fibromatosis (IDF) (n = 2), Lipofibromatosis (n = 1), Fibrous hamartoma of infancy (FHI) (n = 1), Fibromatosis colli (FC) (n = 2) and myofibroma/myofibromatosis (n = 1). FNAC smears were mainly paucicellular with presence of benign spindle shaped cells in a collagenous stroma common to almost all the cases. A few additional findings such as degenerated skeletal muscle fibres, muscle giant cells and mature adipose tissue were also present in some cases. CONCLUSION: Fibroblastic/myofibroblastic tumors although uncommon, form an important category that must be considered in the differential diagnosis of pediatric soft tissue tumors. FNAC cytology features when assessed in a proper clinical setting (specially the age and site of presentation) are helpful in suggesting probable preoperative diagnosis in these lesions.

Renal Pelvic Cholesteatoma: An Uncommon Finding in Children.

Renal cholesteatoma or keratinizing desquamative squamous metaplasia is infrequent in adults and rare in children. We report a case of renal cholesteatoma in a 4-year-old male child who was referred to us as a case of multiple renal calculi with hydronephrosis. We also discuss his management with a review of relevant literature.

Intraoral Foregut Cystic Developmental Malformations: Three cases with a brief review of literature.

Foregut cystic developmental malformations (FCDM) are a type of rare cystic lesion. The occurrence of FCDM is exceedingly uncommon in the intraoral location. We report three cases of FCDM with intraoral location who presented at Chacha Nehru Bal Chikitsalaya, New Delhi, India, in 2016, 2017 and 2018 with symptoms of respiratory distress and feeding difficulties. Two patients were male and one was female with an age range of 29 days to eight years. The clinical differential diagnosis included mucocele, ranula, dermoid, lymphangioma, teratoma, thyroglossal duct cyst, etc. All patients were treated with simple surgical excision and diagnosed, based on histopathology, with FCDM. These should be considered as differential diagnosis of head and neck midline cystic mass lesions. This case report aimed to discuss differential diagnosis and appropriate terminology for these cystic masses as there is varied and ambiguous nomenclature.

Lipofibromatosis.

Introduction: Lipofibromatosis is a benign pediatric soft tissue tumor arising preferentially in the distal extremities. Histologically, the tumor shows abundant adipose tissue admixed with a spindle cell component, often concentrated in septal and perimysial locations. The index case is being presented to discuss the histopathological and immunohistochemical clues to differentiate it from other fibrofatty tumors of childhood.Case report: An 11-month-old male child presented with a slowly growing mass on the upper back. MRI findings were suggestive of an adipocytic tumor. Microscopy revealed a lesion composed of mature adipocytes and intervening fibrous bands with infiltration into the adjacent skeletal muscle, features of lipofibromatosis.Conclusion: Lipofibromatosis should be considered in the differential diagnosis of a pediatric fibrofatty tumor. Accurate diagnosis is essential for proper patient management as incomplete removal of the tumor may result in recurrence.