Childhood cancers: Analysis of 1279 cases at Armed Forces Institute of Pathology, Rawalpindi, Pakistan.

OBJECTIVE: To determine the clinicopathological pattern of childhood malignancies registered with a pathology-based tumour registry. METHODS: The descriptive retrospective study was conducted at the Armed Forces Institute of Pathology, Rawalpindi, Pakistan, and comprised data related to all the histologically diagnosed malignant childhood tumours in the institutional tumour registry from January 2009 to December 2018. Data was analysed using SPSS 20 for the site of involvement, age distribution and histological types of tumours. RESULTS: Of the total 37793 malignant tumours, 1279(3.38%) were in paediatric subjects aged <15 years. There were 820(64.1%) male subjects and 459(35.8%) were female. Lymph node malignancies were the commonest 261(20.4%), followed by eye tumours 251(19.6%), and brain 107(8.3%). CONCLUSIONS: Lymphomas and eye tumours were found to form the main bulk of childhood cancer.

Oxidative stress level and dehydrin gene expression pattern differentiate two contrasting cucumber F1 hybrids under high fertigation treatment.

Two cucumber F1 cultivar hybrids were investigated for stress tolerance markers upon application of different strength of Hoagland fertigation solutions (HG). 'Joker' and 'Oitol' cultivar hybrids were studied, representing typically field grown and greenhouse cultivated genotypes, respectively. At standard fertigation level (0.5 × HG) in controlled environment young 'Joker' plants displayed slower growth than 'Oitol' based on total leaf area. At this basal nutrient concentration leaves of 'Joker' plants had significantly lower antioxidant capacity and higher malondialdehyde (MDA, an indicator of lipid peroxidation) level than 'Oitol'. According to RT-qPCR transcript levels of several antioxidant enzymes' genes (ascorbate peroxidase, glutathione reductase and glutathione peroxidase) were significantly higher in 'Joker' compared to 'Oitol'. At increased HG concentrations (1.0, 1.5, 2.0, and 2.5 × HG) growth didn't change significantly in either hybrid. Osmotic potential declined at higher fertigation levels. Antioxidant capacity increased in both hybrids with strong characteristic differences favouring 'Oitol' plants. Higher MDA content of leaves testified more oxidative burden in 'Joker' plants at all and especially at the more concentrated HG treatments. This trend was also approved by results of bio photon emission imaging, which is a powerful method to quantify stress level in living tissues with autoluminescence detection technology. Gene expression for antioxidant enzymes followed HG concentration-dependent increase in both hybrids, at a substantially higher level in 'Joker'. Expression of the dehydrin gene DHN3 was preferentially induced at elevated fertigation levels in 'Oitol' plants, which could contribute to the lower oxidative stress detected in this hybrid. Results presented in this report demonstrate differences in shoot growth, antioxidant capacity, level of oxidative stress and antioxidant gene expression in two contrasting cucumber hybrids at basal fertigation. Furthermore, excessive HG fertigation was found to increase oxidative stress in a genotype-specific way. This effect may be due to different antioxidant capacity and differential expression of stress protective genes, such as the DHN3 dehydrin.

The Frequency of Double Expresser in Selected Cases of High Grade Diffuse Large B-Cell Lymphomas.

BACKGROUND: Diffuse large B-cell lymphomas (DLBCL) are fast-growing non-Hodgkin lymphomas that affect B-lymphocytes. Double expressor DLBCL is the concomitant expression of Myc and Bcl-2 proteins during lymphomas which results in poor prognosis of patients. This study aimed to determine the frequency of double expresser in high grade diffuse large B-cell lymphomas. MATERIALS AND METHODS: The study was conducted on 74 cases (54 males (68.4%) and 20 females (25.3%)) of DLBCL between August 2018 to January 2019. The mean age of the 74 patients was 51.7 years + 18.5. Expression of proteins c-Myc, Bcl-2 and Bcl-6 were evaluated by immunohistochemistry. The involvement of primary lymph node was reported in 38 cases (51.3%) whereas, extra nodal site was observed in 22 cases (29.7%). Among the primary sites, the cervical lymph node enlargement was the most frequent site of presentation. RESULTS: The rearrangement pattern was studied among 74 patients, 35 (47%) were found to have either one of the rearrangements i.e. Myc, Bcl-2, or Bcl-6. On the other hand, 14 (18.9%) had shown double rearrangements i.e. Bcl-2 and c-Myc (11 cases) and Bcl-6 and c-Myc (3 cases). The Bcl-2 and Bcl-6 rearrangements were demonstrated by 12 cases whereas 2 cases (2.7%) indicated all three types of rearrangements i.e. c-Myc, Bcl-2, and Bcl-6. In 11 cases the Bcl-2 and c-Myc rearrangements were found to be Bcl-2 > 50% and c-Myc > 40% and demonstrating the overall frequency of double expressers as 14.8%. The prognosis of the mentioned cases was extremely poor, median survival of 10 months. CONCLUSION: The concurrent expression of Bcl-2 and c-Myc was found to be 14% (level of expression for Bcl-2 > 50% and c-Myc > 40%) which is potentially a significant health burden and an emerging threat.

Expression of Ki-67 and Βeta-Catenin in Pseudoepitheliomatous Hyperplasia and Squamous Cell Carcinoma in Oral Mucosal Biopsies : An Immunohistochemical Study.

OBJECTIVE: To examine the immunohistochemical expression of Ki-67 and beta-catenin in pseudoepitheliomatous hyperplasia and squamous cell carcinoma (SCC) in oral mucosal biopsies. METHODS: In this comparative cross sectional study, 70 cases of each PEH and OSCC were taken from the patients of both genders and in all age groups. Study was conducted at Armed Forces Institute of Pathology (AFIP), Rawalpindi from Dec 2017 to March 2019. Statistical analysis was done with the help of SPSS Version 24.0. We used Chi-Squared test with p value of < 0.05 which was considered as statistically significant. RESULTS: In the current study, 80 (57.1%) male and 60 (42.8%) female patients with the mean age of 51.69 ± 16.121 (mean ± SD) years were included. It was found that 6-25% Ki-67 labeling index was observed in all (70) PEH cases, which involved only basal layer of the epithelium. Whereas, Ki-67 labeling index was highly expressed in tumor of high grade malignancy than tumor of low grade malignancy. On the other hand, expression of membranous beta-catenin was higher in PEH and cytoplasmic beta-catenin expression was higher in OSCC. CONCLUSION: It is concluded that Ki-67 and beta-catenin showed significant expression in PEH and OSCC in oral mucosal biopsies especially those with intense inflammation or unoriented tissue, helping the clinicians to arrive at a final diagnosis before planning any surgical intervention.

Stromal Expression of CD10 by Immunohistochemistry in Odontogenic Keratocyst (OKC), Dentigerous and Radicular Cysts and Its Correlation with Local Recurrence and Aggressive Behaviour

Objective: To assess and compare the stromal expression of CD10 in OKC, dentigerous and radicular cysts. Materials and Methods: This comparative, cross sectional study was conducted at Armed Forces Institute of Pathology (AFIP), Rawalpindi, from Jan 2017 to Dec 2017. Total sixty cases comprising 20 of each OKC, Dentigerous and Radicular cysts were included in this study. Hematoxylin and eosin (H and E) sections were performed followed by immunohistochemical staining for CD10 antibody. Expression of CD10 was evaluated and compared. Results were analyzed by using SPSS version 20.0. Chi Square test was performed with P value < 0.05 was considered as significant. Results: A total of 60 cases, 20 of each OKC, dentigerous and radicular cysts were taken. In our study, 38 (63.3%) male and 22 (36.7%) female patients with the mean age of 32 ± 15 (mean ± SD) were included. Percentage of CD10 positive cells were highest in sub-epithelial stroma of OKC (95% cases) as compared to radicular and dentigerous cysts (60 and 70%) with highest number of cases showing intense staining in OKC 13(65%) as compared to other odontogenic cysts i-e 4(20%) and 2 (10%) respectively. There was a statistically significant association between odontogenic cysts and proportional score, intensity score and combined score of stromal CD10 expression (P=0.009, p=0.001 and p=0.000). Conclusion: In this study, we found that highest stromal CD10 expression in OKC as compared to dentigerous and radicular cyst, which might be due to aggressive behaviour and increased risk of recurrence in OKC. Expression of CD10 marker will further aid the clinician to plan appropriate surgical intervention and keep regular follow-ups to identify recurrences.

Histopathological and Immunohistochemical Analysis of Small Intestinal Biopsies in Adults Suspected of Celiac Disease.

OBJECTIVE: To analyse histomorphological and immunohistochemical analysis of small intestinal biopsies in adults suspected of celiac disease. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Department of Histopathology, Army Medical College, Rawalpindi, from November 2014 to December 2015. METHODOLOGY: Fifty cases of small intestinal mucosal biopsies (duodenal and jejunal) were analysed in adult patients aged above 14 years suspected of celiac disease. Their histomorphological data was recorded using Modified Marsh Criteria. Type of intraepithelial lymphocytes was assessed using immunohistochemistry. Intraepithelial lymphocytes were counted both by H&E stain and immunostain CD3 and CD20. RESULTS: Thirty-four percent patients were aged between 21 - 30 years and 22% patients aged between 41 - 50 years. There were 84% (n=42) males. Thirteen (26%) cases showed focal villous atrophy, 32 (64%) cases showed partial villous atrophy and 5 (10%) cases showed complete villous atrophy. Anti-tissue transglutaminase antibody was positive in 21 (42%) cases. CD3 immunomarker was positive for intraepithelial lymphocytes in all 50 cases while CD20 immunomarker showed focal positivity in areas with lymphoid follicle formation. The count of intraepithelial lymphocytes was found to be almost equal (with a difference of 3 - 4 lymphocytes) on both H&E stain and immunostain CD3 and CD20. CONCLUSION: Males aged 21 - 30 years were the most commonly affected group. The most frequent change in histology was partial villous atrophy along with lymphocytic enteritis. All the intraepithelial lymphocytes were present in crescendo pattern of distribution.

Endobronchial Ultrasound Guided Transbronchial Needle Aspiration (EBUS-TBNA) for Diagnosis of Mediastinal and Hilar Masses.

OBJECTIVE: To determine the sensitivity, specificity, negative and positive predictive values, and diagnostic accuracy of Endobronchial Ultrasound Guided Transbronchial Needle Aspiration (EBUS-TBNA). STUDY DESIGN: Across-sectional validation study. PLACE AND DURATION OF STUDY: Department of Histopathology, Army Medical College, in collaboration with Department of Pulmonology, Military Hospital Rawalpindi, from March 2014 to March 2015. METHODOLOGY: Cases of EBUS-TBNAcomprised of both TBNAs and cell block/biopsy of the same patients. Diagnosis was made on the TBNAslides and cell block/biopsy material. Taking biopsy/cell block as the gold standard, the data was analysed to calculate the sensitivity, specificity, negative and positive predictive values, and diagnostic accuracy of EBUSTBNA. RESULTS: The sensitivity of EBUS-TBNAwas found to be 96.5%; whereas, specificity and positive predictive values were 100%. The negative predictive value was calculated at 50%. Diagnostic accuracy of the procedure was found to be 96.67%. CONCLUSION: EBUS-TBNA is a sensitive and a specific test and is accurate in diagnosing mediastinal and hilar pathologies.

CD 10 expression intensity in various grades and stages of urothelial carcinoma of urinary bladder.

OBJECTIVE: To evaluate CD10 expression in urothelial carcinoma of the urinary bladder and the association of immunohistochemical (IHC) CD10 expression intensity with grade and stage. STUDY DESIGN: Descriptive cross-sectional analytical study. PLACE AND DURATION OF STUDY: Armed Forces Institute of Pathology, Rawalpindi, from January to December 2011. METHODOLOGY: Fifty consecutive cases of urothelial bladder carcinomas, obtained through transurethral resections, were included in this study. Hematoxylin-eosin (HE) stained sections from each case were re-evaluated histopathologically according to WHO 2004 grading system. The TNM system was used for pathologic staging. On selected slides IHC CD10 marker was applied and a semiquantitative scoring for its expression based on the percentage of positive cells and intensity was performed. Data was entered and analysed on SPSS version 17. Fisher's exact test was used to compare grades, stages of urothelial carcinoma with CD 10 expression and age groups. P < 0.05 was taken as level of significance. RESULTS: Urothelial carcinoma was more common in males. The male to female ratio was 9:1. The older patients > 50 years had higher grade and stage as compared to the younger patients. All cases of high grade urothelial carcinoma showed higher positivity for CD 10. Twenty cases (86.95%) of high grade urothelial carcinoma were positive with +2 immunostaining while 3 cases (13.04 %) were positive with +1 staining. None of the tumors of stage pTa was positive for CD 10 expression. Of all patients with stage pT 1 tumor, 1 case (5.3%) was CD 10 negative and 17 cases (89.9%) were CD 10 positive having +1 staining with 5 - 50% staining and 1 case (5.3%) had +2 staining with more then 50% expression. Out of all patients with stage pT 2, no tumor was CD 10 negative, 3 (13.6%) patients were CD 10 positive with +1 staining and 19 (86.4%) with stage pT 2 tumor had stained positive with +2 staining. CONCLUSION: CD 10 expression was greater in high grade and invasive urothelial carcinomas; it may be associated with tumor progression in bladder cancer pathogenesis.

Changing pattern of malignancies: analysis of histopathology based tumour registry data and comparison of three decades at Armed Forces Institute of Pathology, Rawalpindi, Pakistan.

OBJECTIVE: To assess the pattern of malignancies from tumour registry data and to find any changing pattern by comparing corresponding data from earlier periods. METHODS: The descriptive study was carried out at the Department of Histopathology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, Pakistan. All malignant tumours from 2002-2011 were analysed for age groups, gender distribution, and type of tumour with relation to the site. Tumours of paediatric age group were also assessed separately. Comparison with national and international studies and the data of previous decades - 1977-1988 and 1992-2001 was done to find any changing pattern. SPSS 15 was used for statistical analysis. RESULTS: A total of 32718 malignant tumours were analysed. Of them, males were 19191 and 13527 were females (M:F ratio: 1.4:1). Majority of the patients were between 50-70 years of age. In males, urinary bladder tumours 2153 (11.2%), followed by combined lymphoma/leukaemia 2020 (10.5%) and prostate 1825 (9.5%) were top three malignancies, whereas in females breast 4178 (30.9%) was the commonest. Comparison of different decades showed that in males in the first monograph lymphoma was the commonest, then it was prostatic carcinoma in the second monograph, and now tumours of urinary bladder were on top position. In females, carcinoma cervix, which was at number 5 and 9 in the two earlier monographs was not found in the list of current 10 common tumours. CONCLUSION: The study showed some interesting features, particularly in male malignancies and those related to urinary bladder tumours. The persistent increasing frequency of breast carcinoma in females is also alarming, and requires extensive efforts of awareness, screening and early detection programmes.

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.

Diagnostic challenges and role of immunohistochemistry in metastatic liver disease.

OBJECTIVE: To evaluate the role of Immunohistochemistry (IHC) in the diagnosis of metastatic liver disease, with a descriptive, cross-sectional study at the Department of Histopathology, Armed Forces Institute of Pathology (AFIP), Rawalpindi. MATERIAL AND METHODS: A total of 130 cases of metastatic liver disease were retrieved from the tumor registry data. Eosin-haematoxylin stained sections and Immunohistochemistry panels applied to ascertain the site of primary tumor were evaluated. The panels of detailed immunohistochemical markers were applied. Frequency and percentages were calculated for qualitative variables. Mean and standard deviations were calculated for quantitative variables. RESULTS: Males were 87 (67%) and were females 43 (33.07%). The most common site of primary was in GIT (45%), followed by neuroendocrine carcinoma and gall bladder. The other less common sites were lung, breast, ovary and thyroid. CONCLUSION: There is no specific singular panel of immunohistochemistry markers which can be used in all cases of metastatic liver tumors. The best use and selection of IHC markers depend upon morphological features, clinical history and results of other relevant investigations.

Spectrum of primary non-cutaneous malignant melanomas in northern Pakistan.

OBJECTIVE: To study the frequency and distribution of non-cutaneous malignant melanoma in Northern Pakistan using a cross-sectional descriptive design at the Armed Forces Institute of Pathology Rawalpindi, from 1987-2008. MATERIALS AND METHODS: A total of 234 cases of malignant melanoma reported from 1987-2008 were retrieved from the AFIP tumour registry and analysed for age, gender and site using computer software SPSS 12. RESULTS: Of a total of 58,680 malignant cases reported at AFIP from 1987-2008, only 234 (0.4%) were of malignant melanoma, 142 in males and 92 in females. The age range was 10-97 years, with a mean of 53 ± 16. Only 69 (29.5%) were cutaneous and 165 (70.5%) were non-cutaneous in origin: there were 62 (37.4%) cases from anorectal region, 39 (23.4%)were from eye ball and 17 (10.2%) in the nasopharynx, 13 cases (7.8%) from oral mucosa and 11 (6.6%) from the vagina. CONCLUSION: Malignant melanoma is markedly less cutaneous in Pakistan as compared to the western world, often being found in the anorectal region followed by the eye, nasopharynx, and oral mucosa in descending order.

Diagnostic accuracy of endometrial curettage in endometrial pathology.

BACKGROUND: Abnormal uterine bleeding is one of the most frequent problems in life of an adult female. Uterine curettage or biopsy remains a preferred sampling procedure for diagnosis of the endometrial pathology. The objective of this study was to compare the sensitivity, specificity, positive and negative predictive value of endometrial curettage. PATIENTS AND METHODS: This validation study was carried out at the Department of Histopathology, Army Medical College Rawalpindi in collaboration with Military Hospital, Rawalpindi from January to December 2010. The study included 50 curettage and subsequent hysterectomy specimen of the same patients. Non-probability sampling technique was used to divide patients into two groups with 50 patients in each group. One group was of endometrial curettage having endometrial pathology (group A). Second group was of subsequent hysterectomy specimen of the same patients (group B). RESULTS: Endometrial curettage was found most accurate in diagnosing endometrial carcinoma. Sensitivity of endometrial curettage was found to be 33% whereas specificity and positive predictive value was found to be 100% each. Negative predictive value was found to be 93.1%. CONCLUSION: Endometrial biopsy is a sensitive and a specific test in and is accurate in diagnosing endometrial pathology. It is found most accurate in diagnosing endometrial carcinoma.

Acceptability and accuracy of saline infusion sonohysterography in women with postmenopausal bleeding.

OBJECTIVE: To validate the diagnostic efficacy of saline infusion sonohysterography (SIS) in the evaluation of uterine cavity,in women with postmenopausal bleeding and endometrial thickness (3) 5 mm. STUDY DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Military Hospital, Rawalpindi, from March 2005 to July 2008 and Combined Military Hospital, Multan, from September 2008 to June 2009. METHODOLOGY: Seventy seven eligible participants included women complaining of postmenopausal bleeding were included; out of whom 69 completed the procedure satisfactorily. Pain scores during procedure were assessed to determine patient acceptability. Following saline infusion sonohysterography all patients also underwent an out door pipelle endometrial biopsy in a one-stop postmenopausal bleed clinic. Findings of sonohysterography were compared with hysteroscopy/hysterectomy specimen. RESULTS: Majority of the patients 34 (49.2%) experienced no pain during the procedure, 51 (74%) women had a positive SIS and the findings were negative in 18 (26%) patients. The commonest abnormal finding on SIS was focal thickening in 23 (32%) and endometrial polyp in 12 (17%) cases. Saline infusion hysterosonography picked up 3 cases of false positive polyps (5.8%) and missed a case of submucous fibroid near cervix (1.8%). Upon comparison of findings of hysteroscopy and hysterectomy in 53 cases, there was complete agreement in a total of 43 (88%) cases. The sensitivity of sonohysterography was 92% and specificity was 78%. All the ladies with abnormal SIS had diagnostic or therapeutic procedures and finally only 24 (35%) were managed on conservative follow-up. Normal SIS led to conservative management in 16 (88%). CONCLUSION: Sonohysterography in combination with endometrial biopsy is a useful technique useful for the evaluation of postmenopausal bleeding. Patient acceptability and diagnostic capability is high and it reduces demand for hysteroscopy.

Steroid hormone receptor association with prognostic markers in breast carcinoma in Northern Pakistan.

OBJECTIVE: To determine steroid hormone receptor expression and their association with histological prognostic markers and biological profile in female breast carcinoma in Northern Pakistan. STUDY DESIGN: Cross sectional, observational. PLACE AND DURATION OF STUDY: Department of Histopathology, Armed Forces Institute of Pathology, Rawalpindi, from January 2004 to December 2007. METHODOLOGY: Female patients belonging to Northern Pakistan with primary operable breast carcinoma were included in the study. Patient's age, microscopic tumour size, tumour grade, lymph node status and biological profile for Her-2/neu status were evaluated. Immunohistochemical expression for ER and PR was determined individually and conjointly for ER+PR+ and ER-PR- while their association with above prognostic markers was determined using the c2 test for univariate analysis. RESULTS: Out of the 726 cases, there were 657 (90.4%) cases of infiltrating ductal carcinoma with mean age of 48+/-12 years and mean tumour size of 4.2+/-2.3 cm. Tumour was grade-II in 65% and lymph node metastases in 71.5% cases with Her-2/neu+ in 28.1%. ER and PR expression was 74.6% and 68.3% respectively. Conjoint expression of ER+PR+ was 456 (62.8%), ER-PR- in 21.2%, ER+PR- in 86 (11.8%) and ER-PR+ in 30 (4.1%) cases. Individually, ER and PR showed positive association with age, lymph node metastasis, tumour grade and inverse association with Her-2/neu. Conjoint expression of ER+PR+ (62.8%) and ER-PR-(21.2%) showed positive association with age, tumour grade, lymph node metastasis and inverse association with Her-2/neu (p < 0.05). CONCLUSION: Hormone receptor expression of ER and PR expression is comparable to the West with ER and PR showing inverse association with Her-2/neu and positive association with age, tumour grade and lymph node metastases.

A comparative analysis of coronary adventitial T-lymphocytes--an autopsy study.

BACKGROUND: Recent clinical and histopathologic data suggests that inflammation plays a key role in coronary artery plaque instability and subsequent occlusive thrombosis. The intima has received much attention as a site of inflammation, while the adventitia has remained relatively unexplored. The aim of the present study was to investigate the frequency of inflammatory activity in the cap and shoulder region of unruptured, atherosclerotic lesions in coronary arteries and to correlate these findings with distribution of inflammatory cells in adventitia. METHODS: The study was carried out in Histopathology Department, Army Medical College, Rawalpindi and National University of Sciences & Technology (NUST), from August 2008 to July 2009. Sixty-seven autopsy cases performed at Military Hospital Rawalpindi, Pakistan were selected. The cases were divided into study group and control group. Case group (n = 35) included those where cause of death was ischemic heart disease. Those coronary arteries were taken as control (n = 32) where atherosclerotic changes were found by chance (death without history of ischemic heart disease). Plaques in each group were assessed by light microscopy and by immunohistochemistry. RESULTS: The ages of the deceased ranged from 38 to 49 years. Within study group, adventitial lymphocytes exhibited strong correlation with erosion, thrombus formation in culprit plaque (p = 0.001). No correlation was found between adventitial T-lymphocytes and erosion of plaque (p = 0.700) in control group. In 72% of culprit plaques moderate staining for T-lymphocytes was observed in adventitia as well as intima. In control group, most of the cases contained scattered cells. Few cases of stable plaques revealed lymphocytes as clusters, both in adventitia and in intima. CONCLUSION: Adventitial inflammation may play a pivotal role for atherosclerotic lesion histology and atheroma instability. With the help of these autopsy findings, we hope to be able to reduce the incidence of culprit plaques related to inflammatory reaction in patients of ischemic heart disease.

Morphometric analysis of hepatic steatosis in chronic hepatitis C infection.

BACKGROUND/AIMS: To quantitatively assess steatosis by a morphometric method and to study its relationship with other histological features of chronic hepatitis C (CHC). This was a comparative descriptive study. The study was carried out in the Department of Histopathology, Army Medical College, Rawalpindi, Pakistan, from March 2006 to March 2007. METHODS: Patients who had undergone a liver biopsy for the evaluation of hepatitis C virus (HCV) infection were included in the study. Demographic characteristics and laboratory data were collected at the time of biopsy. The first hundred biopsy specimens that met the inclusion criteria were assessed for grades of steatosis (semiquantitatively), diameter of fat globules (by a morphometric method), necroinflammation, and fibrosis (semiquantitatively). Liver biopsies were processed for paraffin embedding, stained with hematoxylin and eosin, whereas Gomori's Reticulin stain was used for the evaluation of fibrosis. RESULTS: Out of 46 cases showing fatty change, pansteatosis was observed in 24 (52%) patients: 12 (26%) cases had a pericentral and mid zonal distribution of fat globules and eight (17.5%) cases revealed a mid zonal pattern only. There were two (4.5%) cases in which fat globules were found in periportal and mid zonal areas. None of the histological parameters (the stage of fibrosis and grades of inflammation) had any significant correlation with these distribution patterns of steatosis. The diameter of fat droplets was quantified by morphometry. A mixed pattern of steatosis was observed more frequently (21 out of 46 cases): 17 cases had microglobules and eight biopsies showed macroglobules. The size of the fat globules exhibited a significant correlation with the stage of fibrosis (P < 0.0001). The analysis of the grades of necroinflammation did not reveal any significant relationship with the diameter of fat globules. CONCLUSIONS: A mixed pattern of fat globules is more frequently observed in CHC, but macrovesicular steatosis is associated with a higher stage of fibrosis. Morphometry is recommended as one of the important tools for the follow-up of HCV-infected patients. Whether an accurate assessment of fat globule size by morphometry is preferred for the evaluation of patients before and after the antiviral therapy needs further research.

Correlation of steatosis with fibrosis and necro-inflammation in chronic hepatitis C infection in the absence of confounding factors.

OBJECTIVE: To assess the frequency and degree of hepatic steatosis in patients of chronic hepatitis C infection and determine its correlation with stages of fibrosis and necro-inflammatory grades. STUDY DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: The study was carried out in the Department of Pathology (Histopathology), Army Medical College, Rawalpindi, from March 2006 to March 2007. METHODOLOGY: Patients who had undergone a liver biopsy for evaluation of hepatitis C virus infection were included in the study. Demographic characteristics and laboratory data were collected at the time of biopsy. First hundred biopsy specimens meeting the inclusion criteria were assessed for steatosis, necro-inflammation and fibrosis. RESULTS: Steatosis was present in 46 patients and graded as mild (41.3%), moderate (54.3%) and severe (4.3%). An overall significant correlation was found between grades of steatosis with stage of fibrosis (p < 0.0001) but no correlation was found with necro-inflammatory grades. Only focal necrosis revealed weak correlation with grades of steatosis (p < 0.003). CONCLUSION: These results suggest a possible role of the hepatitis C virus itself in the pathogenesis of steatosis and indicate its close relationship with fibrosis and focal degeneration in chronic hepatitis C. Necro-inflammation seen in liver biopsy is host immune reaction to hepatitis C virus and is not related to steatosis.

Frequency of bcl-2 gene rearrangement in B-cell Non-Hodgkin's lymphoma.

OBJECTIVE: The objective of the study was to determine the frequency of bcl-2 gene rearrangement in B-cell Non-Hodgkin's lymphoma (NHL) and identify different breakpoints of bcl-2 gene. METHODS: Thirty cases of B-cell lymphoma (including 8 cases of follicular lymphoma, 19 cases of diffuse large B-cell lymphoma and 3 cases of T-cell rich B-cell lymphoma) were included in the study. Good quality of DNA was extracted in 4 cases from formalin fixed paraffin embedded tissue and in 26 cases from fine needle aspirate. The polymerase chain reaction was done for major break point region (mbr), minor cluster region (mcr) and intermediate cluster region (icr) of the bcl-2 gene. RESULTS: The bcl-2 gene rearrangement was identified in 23.3% of B-cell lymphoma, 50% of follicular lymphoma, 15% of diffuse large B-cell lymphoma and no bcl-2 rearrangement was identified in any of the T-cell rich B-cell lymphomas. Further analysis showed the icr breakpoint in 16.7% of B-cell lymphoma, 37.5% of follicular lymphoma and 10.5% of diffuse large B-cell lymphoma. Involvement of the mbr breakpoint was found in 6.7% of B-cell lymphoma, 12.5% of follicular lymphoma, and 5.3% of diffuse large B-cell lymphoma. Involvement of the mcr breakpoint was not seen in any of the cases. CONCLUSION: The bcl-2 gene rearrangement is quite frequent in follicular lymphoma, followed by diffuse large B-cell lymphoma. The commonest breakpoint in present series is icr followed by mbr. This indicates that primers for bcl-2 gene must include icr primer, whenever the bcl-2 gene is being evaluated for B-cell NHL in this part of the world and this might reduce the variability of frequency of bcl-2 gene rearrangement within and between different regions.

Estrogen receptor, progesterone receptor, HER2/neu, P53 and Ki-67 status of male breast carcinomas in Pakistan.

OBJECTIVE: The aim of present study was to assess the status of estrogen and progesterone receptors, HER2/neu, p53 and ki-67 in male breast carcinomas in our institute. STUDY DESIGN: A descriptive study, carried out in the Department of Pathology (Histopathology), Army Medical College, Rawalpindi, Pakistan from June 2008 to January 2009. SUBJECTS AND METHODS: In this study 45 cases of male breast carcinoma, including all the histological subtypes were assessed with original pathology reports of each case investigated for the age, laterality of breast, histological type of tumour and tumour grade. Tumour blocks of each case were retrieved for immunohistochemical staining of estrogen and progesterone receptors and HER2/neu, ki-67 and p53 scoring was accomplished using established protocols. RESULTS: The majority of the cases were above 65 years of age. Histologically, the invasive ductal carcinoma was the predominant lesion. In total 95.5% of the cases were estrogen and progesterone (ER and PR) receptor positive. The HER2/neu staining was positive in 84.4% cases. According to the percentage of nuclear staining, 77.7% of the cases were p53 positive and 35.6% of the cases had strong nuclear staining intensity. A total of 55.5% of the tumours showed proliferation by ki-67. CONCLUSION: The majority of male breast carcinomas in Pakistan are ER and PR positive and demonstrate immunoreactions for prognostic markers. The results point to a relatively aggressive nature of such lesions in our institute.