Role of Alcian blue (pH 2.5) histochemical stain in diagnostic thyroid pathology: how far is it helpful?

Background: Controversies and pitfalls are well-known in diagnostic thyroid pathology. Since, the luminal surface of  neoplastic thyroid follicular cells is said to be covered by a glycoproteinaceous material, there is a need  to  investigate the efficacy of  mucin stains like Alcian blue (pH 2.5), which can be used as an adjunctive tool in difficult cases.Methods: 60 thyroidectomy specimens, received at Department of Pathology, were included in this retrospective analytical study. Alcian blue histochemical (pH 2.5) staining was done on representative sections and assessed for positivity.Results: Of the 60 cases analysed, all cases of papillary/nodular hyperplasia (n=10), follicular adenoma (n=14), follicular carcinoma (n=4) and medullary carcinoma (n=1) were negative for Alcian blue. Only 3/16 (18.7 %) cases of classical papillary carcinoma and 2/15 (13.3%) cases of follicular variant of papillary carcinoma showed positivity.Conclusions: We got statistically insignificant results with Alcian blue (pH 2.5) staining, making it unreliable in diagnostic thyroid pathology

Utility of horizontal and vertical sections of scalp biopsies in various forms of primary alopecias

BACKGROUND: This study was performed to demonstrate the usefulness of horizontal and vertical sections of scalp biopsies in diagnosing various forms of primary alopecias and to highlight the importance of error-free grossing. MATERIALS AND METHODS: A retrospective analytical review of 228 scalp biopsies was done, noting down the diagnostic histopathological features evident in horizontal and vertical sections of each cases. The idealness of the sections, especially horizontal section, was also analyzed. RESULTS: Out of the 228 cases, 44 scalp biopsies were classified histologically as cicatricial alopecias and the remaining 162 as noncicatricial alopecia. 22 cases were inconclusive owing to erroneous grossing. We found horizontal sections to be more useful in cases of noncicatricial alopecias, whereas vertical sections proved superior in cicatricial alopecias. CONCLUSION: Combining both horizontally and vertically sectioned scalp biopsies maximizes the diagnostic yield. When a single biopsy is submitted, the choice between horizontal and vertical section should depend on clinical diagnosis/suspicion

Reliability of S-100 and Tuj-1 immunofluorescence markers in the diagnosis of Hirschsprung’s disease

Background: Hirschsprung’s disease (HSD) is a congenital malformation of the hindgut characterised by absence of intrinsic ganglion cells. Acetyl cholinesterase (AChE) staining is used as a gold standard test in HSD. Intramucosal neuroglial cells (INCs) are present in aganglionic colonic mucosa of HSD patients, challenging our current understanding.Objectives: To evaluate for the presence of INCs using S100 and Tuj 1 and to evaluate for the presence of ganglion cells (GCs) using Tuj1 immunofluorescence in clinically suspected cases of HSD.Materials and Methods: This study was carried out in the Department of Pathology, Bangalore Medical College and Research Institute, Bangalore on 35 colorectal biopsies of patients suspicious of HSD. Direct immunofluorescence (DIF) was done on rectal biopsies using Tuj1 and S100 markers.Results: Out of 35 biopsies, 24 showed absence of Tuj 1 and S 100 expressing cells in the biopsy and were diagnosed as HSD and 8 cases showed presence of Tuj 1 and S 100 expressing cells in the biopsy and were diagnosed as not suggestive of HSD. 3 were excluded as they were low rectal biopsies. Tuj 1 and S 100 DIF showed sensitivity of 100% and specificity of 72.7%.Conclusion: DIF markers Tuj 1 and S 100 were found to be more sensitive than AChE stain and these markers are helpful in detecting false negative cases missed by AChE. A larger cohort is necessary to consider these DIF markers as an additional tool in evaluatingand ruling out HSD in cases where AChE stain is equivocal

Utility of urine reagent strips in cerebrospinal fluid analysis: An aid to bedside diagnosis of meningitis

Context: The provision of initial treatment to a patient with suspected meningitis depends greatly on early recognition and rapid diagnostic evaluation of cerebrospinal fluid (CSF) leukocytes, proteins, and glucose. The diagnosis is time critical and timely intervention has an implication on the prognosis and outcome. Reasonably, sound laboratorial setups are not available in our country in the primary health-care level and, even in the settings where they are available, long waiting periods precede the availability of results. Aims: We conducted this study to emphasize the role of urine reagent strip test as a rapid diagnostic tool in CSF analysis. Settings and Design: This is a prospective single-blinded study on 100 consecutive CSF samples received with in 1h of tap. Subjects and Methods: All the 100 samples were subjected to definitive test being CSF microscopy and biochemical analysis of proteins and sugar and index test being a semi quantitative analysis of CSF leukocytes, proteins, and sugar by urinary reagent strips. Statistical Analysis Used: The diagnostic accuracy of the reagent strip for different cutoff levels was estimated and tabulated in the form of sensitivity, specificity, positive predictive value, negative predictive value, and likelihood ratio. Results: 77% of cases were in the pediatric age group and 23% cases were adults. The sensitivity and specificity for leukocytes by the strip method for ≥15 cells/cumm were 89.28% and 98.61%, respectively, which increased to 100% with an increase in the counts. The reagent strip test had a sensitivity of 85.71% and a specificity of 95.65% for the protein levels >30 mg/dl which increased to 100% with an increase in protein levels. The reagent strip test for glucose was highly specific (100%) but less sensitive. Conclusions: The results indicate that urine reagent strip is instrumental in bedside CSF analysis and has a future stand in the diagnosis of meningitis

A study to evaluate association of nuclear grooving in benign thyroid lesions with RET/PTC1 and RET/PTC3 gene translocation

Abstract Introduction Papillary thyroid carcinoma (PTC) is the most common malignant lesion of the thyroid characterized by unique histological features like nuclear grooving, nuclear clearing, and intra-nuclear inclusions. However, nuclear grooves are observed even in benign thyroid lesions (BTL) like nodular goiter (NG), Hashimoto's thyroiditis (HT), and follicular adenoma (FA) resulting in diagnostic dilemma of the presence of PTC in such BTL. RET/PTC gene translocation is one of the most common oncogenic rearrangements seen in PTC, known to be associated with nuclear grooving. Among different types of RET/PTC translocations, RET/PTC1 and RET/PTC3 gene translocations are the most common types. These translocations have also been identified in many BTL like hyperplastic nodules and HT. Our study aimed to determine the frequency of nuclear grooving in BTL and evaluate their association with RET/PTC1 and RET/PTC3 gene translocation. Methods Formalin-fixed, paraffin-embedded (FFPE) tissue blocks of NG, HT, and FA were included in the study. The hematoxylin and eosin (H&E) stained sections were evaluated for the presence of nuclear grooving/high power field (hpf) and a scoring of 0 to 3 was used for the number of grooves. Sections of 10 μ thickness were cut and the cells containing the nuclear grooves were picked using Laser-Capture microdissection. About 20 to 50 such cells were microdissected in each of the cases followed by RNA extraction, cDNA conversion, realtime-polymerase chain reaction (RQ-PCR) for RET/PTC1 and RET/PTC3 gene translocation, and the findings were analyzed for statistical significance. Results Out of 87 BTL included in the study, 67 (77.0%) were NG, 12 (13.7%) were HT, and 8 (9.2%) were FA. Thirty-two cases (36.8%) had nuclear grooving with 18 out of 67 NG, 6 out of 12 HT, and all 8 cases of FA showing a varying number of nuclear grooves. A significant association between the number of nuclear grooves with RET/PTC gene translocation (p-value of 0.001) was obtained. A significant association of HT with RET/PTC gene translocation (p-value of 0.038) was observed. RET/PTC1 and RET/PTC3 translocation were seen in 5 out of 87 cases, with HT showing positivity in 2 and FA in 1 case for RET/PTC1 and HT in 1 and FA in 2 cases for RET/PTC3 gene translocation with 1 case of FA being positive for both RET/PTC1 and RET/PTC3 gene translocation. Conclusions The frequency of nuclear grooving among BTLs in our study was 36.8%. Our study shows, that when BTLs, show nuclear grooves, with an increase in the nuclear size, oval and elongated shape, favors the possibility of an underlying genetic aberration like RET/PTC gene translocation, which in turn supports the reporting pathologist to suggest a close follow up of the patients on seeing such nuclear features on cytology or histopathology sample, particularly in HT