Mulazim Hussain Bukhari

An updated audit of fine needle aspiration cytology procedure of solitary thyroid nodule.

This study was conducted to see the sensitivity, specificity, and accuracy of fine needle aspiration cytology (FNAC) for solitary thyroid nodules and to compare our experience with that of other regions of the world. It was a prospective cross sectional study conducted on 76 thyroid samples submitted and reported at the Department of Pathology, King Edward Medical University, Lahore. There were 65 (85.5%) females and 11 (14.5%) males. Male to female ratio was 1:5.9. Ages of the patients ranged from 10 to 60 year with mean age 35.37 ± 12.17. Thirty‐three (43.42%) samples were indeterminate. There were 30 cases (39.47%) of benign lesions, comprising of colloid goiter, follicular adenoma, and diffuse hyperplasia. The final reports of these cases were almost the same. On FNAC, 13 cases were declared as malignant (6 cases) or suggestive of malignancy (3 cases) or suspicious for malignancy [4 cases (5.26%)]. Only 9 cases (11.84%) were clearly committed as malignant lesions, comprising of papillary carcinoma, anaplastic carcinoma and suggestive of follicular carcinoma. Comparison of malignant cases on histopathology (14 cases) was close to that of FNAC (13 cases). After comparison of FNAC results with histopathology, overall sensitivity of FNAC was found to be 90%, specificity 87.5%, and accuracy 87%, while positive predictive value (PPV) was 93% and negative predictive value (NPV) was 79.5%. In conclusion, we recommend this procedure in the light of views of other experts as a primary investigation of thyroid lesions. We strongly recommend the suggestion that in a patient with one or more thyroid nodule, FNAC should be advised for every patient for exclusion of cancer. We will also encourage the clinicians to embrace this procedure in the initial management of patients. As FNAC is inexpensive, sensitive, specific, and an accurate procedure it should be adapted as an initial investigation of thyroid diseases in all tertiary hospitals in developing countries like Pakistan. Diagn. Cytopathol. 2008; 36:104–112.

Human papillomavirus in upper digestive tract tumors from three countries

AIM To clarify human papillomavirus (HPV) involvement in carcinogenesis of the upper digestive tract of virological and pathological analyses. METHODS The present study examined the presence of HPV in squamous cell carcinomas of the oral cavity (n = 71), and esophagus (n = 166) collected from Japan, Pakistan and Colombia, with different HPV exposure risk and genetic backgrounds. The viral load and physical status of HPV16 and HPV16-E6 variants were examined. Comparison of p53 and p16(INK4a) expression in HPV-positive and HPV-negative cases was also made. RESULTS HPV16 was found in 39 (55%) oral carcinomas (OCs) and 24 (14%) esophageal carcinomas (ECs). This site-specific difference in HPV detection between OCs and ECs was statistically significant (P < 0.001). There was a significant difference in the geographical distribution of HPV16-E6 variants. Multiple infections of different HPV types were found in 13 ECs, but multiple infections were not found in OCs. This difference was statistically significant (P = 0.001). The geometric means (95% confidence interval) of HPV16 viral load in OCs and ECs were 0.06 (0.02-0.18) and 0.12 (0.05-0.27) copies per cell, respectively. The expression of p16(INK4a) proteins was increased by the presence of HPV in ECs (53% and 33% in HPV-positive and -negative ECs, respectively; P = 0.036), and the high-risk type of the HPV genome was not detected in surrounding normal esophageal mucosa of HPV-positive ECs. CONCLUSION Based on our results, we cannot deny the possibility of HPV16 involvement in the carcinogenesis of the esophagus.

Diagnostic reliability of FNAC for salivary gland swellings: A comparative study

A prospective study was conducted to see the sensitivity, specificity, and accuracy of fine needle aspiration cytology (FNAC) for 100 salivary gland swellings in comparison with biopsy. These randomized samples were submitted and reported at the department of pathology, Allama Iqbal Medical College, Lahore. The male to female ratio was 1:1.5. Ages of the patients ranged from 8.5 to 58 years with mean age 33.39 ± 12.37 years. Maximum number of lesions was found in age group between 21 and 40 years. Parotid gland was involved in 68%, submandibular gland in 30%, and minor salivary glands in 2% patients. Sublingual gland was not involved in any of our subjects. There were 14% cases of non‐neoplastic lesions and 86% cases of neoplastic lesions on biopsy. Non‐neoplastic lesions included 12 cases of inflammations (three cases of granulomatous inflammation and nine cases of nonspecific chronic inflammation) and two cases of inclusion cysts. Among neoplastic lesions, biopsy revealed 68 cases of benign neoplasia and 18 cases of malignant neoplasia. Non‐neoplastic lesions did not show any difference in diagnosis by both techniques. FNAC misdiagnosed four malignant and one benign lesion. Sensitivity, specificity, positive predictive value, and negative predictive value of FNAC for benign neoplastic lesions were 98.52, 87.05, 94.36, and 96.55%, respectively, whereas for malignant neoplastic lesions they were 77.77, 98.78, 93.33, and 95.29%, respectively. In conclusion, FNAC is found to be a highly sensitive and specific technique for diagnosis of most of salivary gland swellings, except for malignant neoplastic lesions where its sensitivity is intermediate. We strongly recommend that FNAC should be adopted as an initial investigation for all salivary gland swellings. Diagn. Cytopathol. 2010.

Modified method of AgNOR staining for tissue and interpretation in histopathology.

This study was conducted in the department of Pathology King Edward Medical University, from June to December 2002 to introduce the new method of AgNOR staining and its interpretation to increase its reliability. A total of 60 brain specimens were stained with modified AgNOR technique. The diagnosis of malignancy was made on H & E staining. AgNOR counts, variation in size and dispersion of AgNOR dots in cells were graded and compared in malignant and non‐malignant lesions. Modified method of AgNOR staining and interpretation was an easy, reliable and reproducible alternative to traditional AgNOR techniques for evaluating proliferation activity of cells in malignant and benign brain lesions. mAgNOR counts of different grades of astrocytoma (2.97 ± 0.96, 3.97 ± 0.43, 6.01 ± 2.74 and 8.01 ± 3.56) were significantly (P < 0.01) greater when compared with counts of normal brain (0.40 ± 0.01), and reactive gliosis (0.60 ± 0.01). AgNOR size and dispersion were of higher grade in a significantly greater proportion of malignancy when compared with benign conditions (P < 0.05). The AgNOR dots were brighter and more clear with modified staining when compared with previous studies. We conclude that modified AgNOR staining technique is simple, quick and reliable to evaluate cell proliferation by detecting AgNORs size and dispersion. In future, AgNOR size and dispersion should be considered rather than the count only. We recommend the use of morphometry for AgNOR size in future. We also recommend the use of modified AgNOR staining for obtaining sound and confidant results in routine paraffin sections.

Comparison of Fine Needle Aspiration Cytology and Thyroid Scan in Solitary Thyroid Nodule

Objective. This was a comparative study between FNAC and thyroid scan used to diagnose the solitary thyroid nodule and histopathology was used as gold standard to compare the results of both modalities. We hypothesized that Fine needle aspiration cytology and thyroid scan diagnose solitary thyroid nodule (STN) as accurately as histopathology. Materials and Methods. This study comprised of 50 patients with solitary thyroid nodules (STN) presented to OPD. After clinical examination these patients were referred to Centre for Nuclear Medicine, Mayo Hospital Lahore for thyroid function tests and thyroid scan (TS). These patients underwent FNAC in the department of Pathology and surgery in Mayo Hospital. The cases were operated and evaluated for histopathological changes. Results. On thyroid scan, 40 patients (80%) having cold nodule were labeled as suspicious 10 patients (20%) had hot nodule. On FNAC 23 patients (46%) had benign lesion, 22 patients (44%) had indeterminate lesion and 5 patients (10%) had malignant lesions. On histopathology, 45 patients (90%) were confirmed to have benign lesions and 5 patients (10%), malignant lesions. After comparison of results of thyroid scan and FNAC with histopathology, the sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of thyroid scan were 80%, 20%, 10%, 90% and 26%, respectively whereas those of FNAC were 80%, 97.7%, 80%, 97.7% and 96%, respectively. Conclusion. Fine needle aspiration was a significantly better predictor of malignancy than thyroid scan and resulted in a smaller proportion of excisions for benign nodules.

Clinicopathological importance of Papanicolaou smears for the diagnosis of premalignant and malignant lesions of the cervix

Background: Premalignant and malignant lesions are not uncommon in Pakistani women, especially in the older age-groups Aim: This study was conducted to determine the clinicopathological importance of conventional Papanicolaou (Pap) smears for the diagnosis of premalignant and malignant lesions of the cervix. Materials and Methods: Pap smears of 1000 women were examined from January 2007 to June 2009. Only cases with neoplastic cytology were included. Results: The overall frequency of normal, inadequate, neoplastic, and infective smears was 50%, 1.8%, 10.2%, and 38.3%, respectively. Most of the patients (67%) were in the postmenopausal age-group, with the mean age being 44.7±15.63 years. The commonest clinical signs/symptoms seen among the 102 patients with neoplastic gynecological lesions were vaginal discharge and abnormal bleeding (93/102;(91.2% and 62/102;60.7%). Of the 102 cases with neoplastic lesions 46 patients (45%) had low-grade squamous cell intraepithelial lesions (LSILs), 22 (21.5%) had high-grade squamous cell intraepithelial lesions (HSILs), 14 (13.7%) had squamous cell carcinoma, and 6 (5.8%) showed features of adenocarcinoma. Ten (9.8%) cases showed cytology of atypical squamous cells of undetermined significance (ASCUS) and four (3.9%) cases had atypical glandular cells of undetermined significance (AGUS). Conclusion: We conclude that cervical smear examination is well suited for diagnosing neoplastic disease. It is clear that cervical neoplastic lesions are becoming a problem in Pakistan.

Use of Fine-Needle Aspiration in the Evaluation of Breast Lumps

Background. A study was designed to see the role of fine needle aspiration cytology (FNAC) in palpable breast lumps. Materials and Methods. Four hundred and twenty five (425) patients came to the Department of Pathology King Edward Medical University, Lahore in four years for FNAC of their palpable breast masses from June 2006 to June 2010. FNAC diagnosis was compared with histological diagnosis to see the accuracy of fine needle aspiration cytology for neoplastic lesions. Results. There were 271/425 benign, 120/425 malignant, and 32/425 suspicious smears. Inadequate samples were repeated twice or thrice, and the degree of success was improved with consecutive repeating approaches. The frequency of inadequacy declined from 86 to 18, and 2 for first, second and third attempts, respectively. The number of repeats increased the diagnostic accuracy of aspirates which is statistically significant (P = .000). Invasive ductal carcinoma was the most commonly reported lesion with maximum incidence in the 4th, 5th, and 6th decades followed by invasive lobular carcinoma and other malignant lesions. The sensitivity, specificity, accuracy, negative predictive value, and the positive predictive value of FNAC was 98%, 100%, 98%, 100%, and 97%, respectively. Conclusion. FNAC serves as a rapid, economical, and reliable tool for the diagnosis of palpable breast lesions because the cytopathological examination of these lesions before operation or treatment serves as an important diagnostic modality.

Clinicopathological study of Papanicolaou (Pap) smears for diagnosing of cervical infections

Cervical infections are not uncommon in our population especially in young and sexually active women. One thousand samples of married women, aged between 20 and 70 years, were studied by conventional Papanicolaou smears. These samples were examined in the Department of Pathology, King Edward Medical University, Lahore from January 2007 to June 2009. Only cases without (pre)neoplastic cytology were included. Six types of infections were diagnosed cytologically. The overall frequency of normal, inadequate, neoplastic, and infective smears was 50%, 1.8%, 10.2%, and 38.3%, respectively. Most of the patients (67%) were in the reproductive age group with mean age 34.7 ± 2.6 years. The commonest clinical sign seen in 354/383 (92%) cases and symptom in (349/383; 91%) cases were vaginal discharge and pruritis vulvae. Among the infective smears, 290 cases (75.7%), the cytologic diagnosis was nonspecific inflammation. Most of these 290 smears contained clue cells (indicating Gardnerella infection) and a lack of lactobacilli. Such smears are predominant in patients suffering from bacterial vaginosis (BV). Twenty‐eight smears (7.3%) were positive for Trichomonas vaginalis, 27 cases (7%) were smears with koilocytic change pathognomonic of human papilloma virus infection. Twenty‐five smears (6.5%) were positive for fungal infection. Seven cases (1.8%) were diagnosed as herpes simplex virus infection. Finally, there were six cases (1.5%) with atrophic vaginitis. We conclude that the cervical smear is well suited for diagnosing cervical infections. It is clear that Gardnerella, known to be associated with bacterial vaginosis, is a major problem in our Pakistani population. Diagn. Cytopathol. 2012.

Evaluation for Granulomatous Inflammation on Fine Needle Aspiration Cytology Using Special Stains

Background. Tuberculosis is the commonest infectious disease in the developing world. Many diagnostic tests are devised for its detection including direct smear examination. This study was designed to determine the frequency of cases positive for AFB and positive for fungus in patients diagnosed to have granulomatous inflammation on Fine Needle Aspiration Cytology using special stains. Materials and Methods. A descriptive cross-sectional survey was done on 100 cases of granulomatous inflammation consistent with tuberculosis diagnosed on fine needle aspiration cytology at the Department of Pathology, King Edward Medical University, Lahore. After reporting granulomatous inflammation on Hematoxylin & Eosin staining of aspirates from FNAC, some unstained slides were subjected to special stains, like ZN, GMS, and PAS. Cases positive for AFB on ZN stain and fungus on GMS/PAS were noted down along with their frequency and percentages. Results. Forty-four cases (44%) of AFB positive smears were reported in granulomatous inflammation while only 5% cases of fungus were reported down. Cervical lymph nodes were the most commonly involved site (87%), and females were affected more (62%) than males. Most cases of AFB-positive smears were associated with caseation necrosis (93%). Conclusion. Special stains should be done on all granulomatous inflammation cases seen on FNAC for confirmation of TB and ruling out other infectious causes.

Elevated frequency of p53 genetic mutations and AgNOR values in squamous cell carcinoma

Background:  Epidermal squamous cell carcinoma (SCC) is a common malignancy in Pakistan. We hypothesize that it is characterized by higher frequency of p53 genetic mutations and increased AgNOR values compared with squamous cell papilloma (SCP) and basal cell carcinoma (BCC).