Down Regulation of KAI1/CD82 in Lymph Node Positive and Advanced T-Stage Group in Breast Cancer Patients.

BACKGROUND: Metastasis represents a deadly aspect of any cancer including breast cancer, given its high prevalence; treatment of metastatic breast cancer remains a clinically unmet need, which necessitates the exploration of metastasis suppressor genes (MSGs). KAI-1/CD82 is an important member of MSGs; the role of KAI1 has been well explored in prostate cancer, however its role in breast cancer is not fully explored and in fact the results of breast cancer studies are contentious. Thus, the present study aimed to investigate expression of KAI1 at both transcriptional and translational levels in the tissue of breast cancer patients and benign breast disease. Further, we analysed the relationship between expression levels of KAI1 and clinicopathological parameters in breast cancer patients. MATERIALS AND METHODS: mRNA expression was studied by Real time PCR and protein expression was analyzed by both Western blot and Immunohistochemistry. RESULTS: The results of the study indicate that KAI1 expression was remarkably decreased in breast cancer both at the gene and the protein levels (P < 0.05) compared to benign breast disease. In addition, KAI1 expression levels were strongly associated with axillary lymph node status and advanced T stage (p < 0.05), however no association was found with tumor grade, age, menopausal status and receptor status like ER, PR and Her2. CONCLUSION: Low expression of KAI1 might be helpful for predicting the lymph node metastasis and T staging, thus predicts malignant prognosis of breast cancer.
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p16 promoter methylation, expression, and its association with estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 subtype of breast carcinoma.

OBJECTIVES: The purpose of the study is to investigate p16 protein expression and promoter methylation of p16 gene and their association with molecular subtypes based on parameter such as estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). MATERIALS AND METHODS: A total of 114 breast cancer tissue biopsies were collected for methylation-specific polymerase chain reaction (MSP) and immunohistochemical (IHC) analysis. RESULTS: Seven tissue microarrays were constructed. p16 protein expression was studied in 114 cases, of which 35/114 (30.7%) cases showed strong expression and the majority of them had ER-positive tumor (57.6%), and it was statistically significant (P < 0.0074). Similarly, p16 expression was reduced in the majority of PR-negative tumors (83.9%) and the association was statistically significant (P = 0.0026). p16 methylation was studied in 114 cases and was positive in 71.0% cases. CONCLUSION: High p16 protein expression was associated with ER-positive, PR-negative, and HER2-negative tumors which is associated with poor prognosis. p16 protein expression may be used as a prognostic indicator to predict treatment response to hormonal therapy.

Y-Box binding protein 1 expression in breast cancer and its correlation with hormone receptors and other prognostic markers.

INTRODUCTION: The present histologic and immunohistochemical prognostic markers of breast carcinoma do not effectively identify the subset of patients with poor prognosis. Y-Box binding protein 1 (YB1) is a novel biomarker which may identify and aid in targeted personalized therapy for such patients. MATERIALS AND METHODS: The study was conducted on histopathology specimens of 74 patients of breast carcinoma who had undergone modified radical mastectomy. YB1 immunohistochemistry (IHC) was performed on manual tissue microarray blocks (each having 15 cores). The YB1 expression was quantified in terms of "immunoreactive score" which was correlated with clinical parameters, hormone receptor status, and Her2neu overexpression by IHC. The Her2neu status of the equivocal cases was further evaluated by fluorescent in situ hybridization (FISH). RESULTS: YB1 was positive in 36/74 (48.6%) cases. On IHC and analysis by FISH, 25/74 (34%) cases had Her2neu overexpression. Estrogen receptor (ER) and progesterone receptor (PR) positivity was found in 42% and 36.5% cases, respectively. YB1 immunopositivity was negatively correlated with ER and PR expression, but showed a significant positive correlation with Her2neu expression. No correlation was found with other clinical parameters, tumor stage, and grade, except lymph node involvement, which showed a positive association with YB1 expression. Triple-negative breast carcinoma constituted 25.6% of the total cases, out of which 73.6% were YB1 positive. CONCLUSION: This study found that YB1 has an association with Her2neu expression. It may in future provide a therapeutic target in Her2neu overexpressing tumors.

BRCA1 Promoter Methylation and Expression - Associations with ER+, PR+ and HER2+ Subtypes of Breast Carcinoma

Introduction: Considering the increasing trend in incidence rates, morbidity and mortality of breast cancer, there is an urgent need to identify and validate new biomarkers for early detection and better management. The purpose of the study was to investigate BRCA1 protein expression and promoter methylation of the BRCA1 gene and their association with molecular subtypes based on estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) positivity. Materials and Methods: A total of 114 breast cancer tissue biopsies were collected for methylation specific PCR (MSP) and immunohistochemical (IHC) analysis. Results: Seven tissue microarrays were constructed. BRCA1 protein expression was reduced in 55/114 (48.2%) and in the majority of ER-negative tumors (73.3%) (p<0.001). Similarly BRCA1 expression was reduced in the majority of PR-negative tumors (69.2%) but without statistical significance (p value=0.083). BRCA1 methylation was positive in 59.6% cases. A subset regarding ER+, PR+ and HER2+ was identified which consisted of 31.6% in which an inverse relationship between BRCA1 methylation and protein expression was noted. Conclusion: Reduced expression was associated with ER and PR negative status which is linked with a poor prognosis. BRCA1 protein expression might thus be used as a prognostic indicator to predict treatment response to hormone therapy.

A Comparison of Vertical and Transverse Sections in the Histological Diagnosis of Alopecia Areata Scalp Biopsy Specimens.

CONTEXT: Both vertical and transverse sections are used for the diagnosis of alopecia areata. However when a single biopsy is submitted the pathologist has to decide which type is better. AIMS: To compare the diagnostic histological features in vertical and transverse sections in alopecia areata scalp biopsy specimens. SETTINGS AND DESIGN: Tertiary Care Hospital. Comparative Study. MATERIALS AND METHODS: A total of 30 patients were enrolled in the study. Two four mm punch biopsy were taken. One was used to take vertical sections and the other for transverse section and histological features of alopecia areata noted in both. STATISTICAL ANALYSIS USED: Chi-square test, percentage. RESULTS: Diagnosis of alopecia areata could be made in 30 (100%) cases in transverse sections and 28 cases (93.3%) in vertical sections. The number of hair follicles available for evaluation was more in the transverse section. Nanogen follicles and miniaturization of follicles were better visualized in the transverse sections. However the catagen and telogen follicles were noted in both vertical and transverse sections but the number and the ratio of anagen and telogen hair follicles could be better assessed in the transverse sections. Presence of peribulbar lymphocytic infiltrate, eosinophils and pigment casts were noted in both transverse and vertical sections. CONCLUSIONS: Transverse sections provide a better assessment of the histological features of alopecia areata than vertical sections and thus should be preferred.

Frontal Bone Hemangioma in an 8-year-old Female: A Common Tumor in a Rare Location.

Intraosseous hemangioma is a rare bone tumor accounting for 0.7%-1.0% of all bone tumors. In the skull, frontal bone is the commonly involved bone. An 8-year-old female presented to our outpatient department with complaints of pain and swelling over forehead for 4 months. X-ray revealed a lytic expansile lesion involving frontal bone with sunburst pattern of bony spicules radiating to periphery of the lesion. Magnetic resonance imaging revealed the presence of a well-circumscribed lesion with both intra as well as extracranial components. Histopathology revealed a vascular tumor consisting of both small (capillary) and large (cavernous) sized vessels. A diagnosis of mixed type of hemangioma of the frontal bone was given. Recognition of hemangioma on radiology and confirmation by histopathology is essential for proper management as it might be confused clinically with other locally aggressive/malignant lesions.

Assessment of prophylactic effect of Senna auriculata (L.) Roxb. leaves on alcohol-induced pancreatitis in rat model.

Aqueous leaf extract of Senna auriculata (L.) Roxb. syn. Cassia auriculata (SLEx) is known to possess potential antidiabetic and antioxidant properties. Based on the known correlation between exocrine pancreatic function and endocrine secretary capacity, here, we studied the prophylactic effect of the SLEx on alcohol induced pancreatitis in rats. To induce chronic pancreatitis, the rats were fed with unsaturated fat i.e. corn oil (2.5 mL/kg) along with high dose of ethanol (10.2 g/kg) for 4 wk, and was increased 0.6 g/kg after every 2 days for 1 wk and then 0.6 g/kg after every 4 days for a period of 4 wk. SLEx was orally administered to rats at dose of 400 mg/kg/day for 4 wk. At the end of 4th wk, pancreatic enzymes i.e., α-amylase, lipase, serum and pancreatic MDA levels were estimated. Pancreatic histopathological studies were also performed. The SLEx significantly reduced the serum levels of α-amylase and lipase along with significant suppression in serum and pancreatic tissue lipid peroxidation. Histomorphological studies did not show any fatty vacoules in acinar cells of SLEx-treated rats. However, vacoulation was seen in acini of pathogenic control rats. With the results, we conclude that Senna auriculata aqueous leaf extract has potential to reduce the ethanol-induced pathogenecity, and it possesses prophylactic effect on alcohol-induced pancreatitis. However, a long term trial is needed to ascertain its therapeutic potential for pancreatitis.

Expression of BRCA1 and BRCA2 proteins and their correlation with clinical staging in breast cancer.

OBJECTIVES: The purpose of this study was to evaluate the level of expression of the BRCA1 and BRCA2 proteins in sporadic breast cancer cases to determine the functional role of these genes in breast carcinogenesis. MATERIALS AND METHODS: Paraffin embedded histologically proven invasive breast tissue sections that were obtained from 40 patients and the adjacent normal breast tissue sections used as controls to determine breast carcinoma specific changes in the expression of BRCA1 and BRCA2 by immunohistochemistry (IHC). RESULTS: Majority of the cases express either low or no detectable level of BRCA1 expression in tumor tissues in comparison with control; the decline in BRCA1 expression was found to be more prominent in advanced grade 3 disease. On the other hand, the expression of BRCA2 protein was moderate or low in breast cancer cases and its overall distribution did not show significant difference when compared with controls. Interestingly, those breast cancer cases, which were found to express low or no BRCA1 expression, demonstrated a higher protein level of BRCA2. The inverse correlation of BRCA1 and BRCA2 expression was more prominent in post-menopausal patients. CONCLUSIONS: Our results demonstrate in a subset of cases that decline in BRCA1 expression that may be associated with potentially compensatory increase in BRCA2 protein, which may depend on tumor grade as well as menopausal status.

Correlation of mitotic indices, AgNor count, Ki-67 and Bcl-2 with grade and stage in papillary urothelial bladder cancer.

PURPOSE: To evaluate the mutual inter-relationship of mitotic indices, argyrophilic nuclear organizer regions (AgNOR) count, Ki-67 and B-cell lymphoma 2 protein (bcl-2) in papillary urothelial bladder cancer (pUBC), and their correlation with grade and stage. To establish the cut-off values of these markers to detect high grade and muscle invasive bladder cancer. MATERIALS AND METHODS: Fifty-four patients with primary pUBC who underwent transurethral resection / radical cystectomy were analyzed retrospectively. Cell proliferation was assessed by Ki-67 labelling index, mean AgNOR count, mitotic count, mitotic activity index and mitosis/ volume index. Immunohistochemistry was done to see bcl-2 and Ki-67 expression. Correlation of these indices with tumor grade and stage and amongst themselves was assessed. The receiver operating characteristic (ROC) curves were drawn to establish the cut-off values. RESULTS: We found a strong positive correlation of mitotic indices and Ki-67 with tumor grade (P = .000), stage (P < .05) and bcl-2 (P = .000). AgNOR count correlated positively with the grade (P = .006), mitotic indices and Ki-67 (P = .032) but not with tumor stage and bcl-2. Cytoplasmic bcl-2 immunopositivity was seen in 42.3% of low grade pUBC and 85.7% of high grade pUBC cases (P = .001). bcl-2 positivity was seen in 85% of muscle invasive pUBC as compared to only 52.9% of superficial cases. Ki-67 ≥ 32.5%, ≥ 14 mitoses/10 high power fields (hpf), ≥ 11.20 mitoses/mm2, ≥ 0.75 mitoses/100 tumor cells and AgNOR ≥ 11.55 are 100% specific for high grade bladder carcinoma. Ki-67 ≥ 59% and mitoses ≥ 36.50 per 10 hpf can indicate muscle invasion with 100% specificity. CONCLUSION: Cut-off values for Ki-67, mitotic indices and AgNOR can confirm high grade bladder carcinoma in equivocal cases. Ki-67 and mitotic count can serve as potential and reliable indicators of muscle invasion.

Comparative evaluation of bone marrow aspirate with trephine biopsy in hematological disorders and determination of optimum trephine length in lymphoma infiltration.

INTRODUCTION: Bone marrow examination is an indispensable diagnostic tool to evaluate neoplastic and non neoplastic hematological diseases. AIMS: To compare bone marrow aspirate with trephine biopsy in hematological disorders. To determine the optimum trephine preprocessing length in lymphoma infiltration. METHODS: Diagnostic comparison was done between simultaneous bone marrow aspirates and trephine biopsies in 449 patients. Biopsies were fixed in formalin, decalcified in 5.5% EDTA and routinely processed. Concordance rates and validity parameters for aspirate were calculated. Three deeper sections of trephine biopsy, cut at 0.1-0.2 mm intervals, were assessed for lymphoma involvement. Proportion of biopsies showing marrow infiltration by lymphoma cells was plotted against trephine length and correlation was assessed. RESULTS: Aspirate had a high sensitivity for acute leukemia (89.4%) and multiple myeloma (88.5%), moderate for NHL (67.6%) and nonhematopoietic metastases (58.3%) and low for aplastic anemia (38.5%) and Hodgkin lymphoma (5%). Aspirate has no role in granulomatous myelitis and myelofibrosis. Lymphoma positivity increased with trephine length, with maximum positivity (68.9%) seen in 17-20 mm group and no further gain beyond 20 mm. (lymphoma positivity ≤16mm=40.3% and ≥17mm=66.1%, p=0.0011). CONCLUSION: Aspirate has a high specificity; its sensitivity depends upon the type of disease. Apart from few conditions, in which aspirate alone is sufficient, biopsy is mandatory in most. Preprocessing trephine length of 17-20 mm examined at multiple deeper levels was found optimal for assessing lymphoma positivity.

Eugenia jambolana pretreatment prevents isoproterenol-induced myocardial damage in rats: evidence from biochemical, molecular, and histopathological studies.

Preventive effects of hydroalcoholic extract of fruit pulp of Eugenia jambolana (HEEJ) on isoproterenol (ISP)-induced myocardial damage in rats were evaluated. Rats were pre-treated with HEEJ (100, 200, and 400 mg/kg) daily for 30 days. ISP (85 mg/kg bw) was administered on the 28th and 29th days at an interval of 24 h. Ischemic control group exhibited significant increases in oxidative stress parameters, markers of inflammation, cardiac damage markers, and apoptotic markers. Oral pre-treatment with HEEJ (100, 200, and 400 mg/kg bw) provided cardioprotective activity by decreasing levels of malondialdehyde, cardiac markers (serum glutamate oxaloacetate transaminase, creatine kinase-myocardial band, cardiac troponin I), and markers of inflammation (interleukin-6, C-reactive protein, and tumor necrosis factor alpha); and increased levels of superoxide dismutase and reduced glutathione. HEEJ (400 mg/kg bw) was found to exert significantly greater effects in comparison to HEEJ (100 and 200 mg/kg bw). Apoptotic marker Bcl-2 was increased, while Bax was decreased in pre-treated rats, which was further confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. The present study provides evidence that pre-treatment with HEEJ attenuates oxidative stress, apoptosis and improves cardiac architecture in ISP-induced rats and, hence, is cardioprotective.

Pleomorphic variant of lobular carcinoma breast: a rare case report with review of the literature.

Pleomorphic carcinoma is a poorly described entity whose phenotype is not well recognized as within the morphological spectrum of breast carcinoma. The purpose of this report is to describe the clinicopathological features of this tumour with review of the literature. We report a case of invasive pleomorphic lobular carcinoma with coexisting classic lobular carcinoma in situ.

Cockayne syndrome-xeroderma pigmentosum complex with demyelination: A rare association.

Xeroderma pigmentosum-Cockayne syndrome (XP-CS) includes facial freckling and early skin cancers typical of XP and some features typical of CS, such as mental retardation, spasticity, short stature, and hypogonadism. XP-CS does not include skeletal involvement, the facial phenotype of CS, or CNS demyelination and calcifications. We present a rare patient whose genome probably harbored a specific combination of mutations producing a rare double syndrome of XP-CS, with facial phenotype of CS, and CNS demyelination.

Salutary effect of Cassia auriculata L. Leaves on hyperglycemia-induced atherosclerotic environment in streptozotocin rats.

Diabetes mellitus is very often associated with dyslipidemia, increased oxidative stress and endothelial dysfunction that could develop atherosclerosis and consequently cardiovascular diseases. Medicinal plants with reputed traditional use to treat diabetes and cardiovascular diseases might provide valuable drugs. Therefore, the present study was designed to evaluate anti-atherosclerotic potential of aqueous extract of Cassia auriculata L. leaves in streptozotocin (STZ)-induced diabetic rats. The rats were rendered diabetic by STZ (45 mg/kg, ip). Diabetic rats were orally administered C. auriculata leaf extract at 400 mg/kg dose daily for 21 days. The supplementation of extract to the diabetic rats produced significant reduction in fasting blood glucose along with significant reversal in altered serum lipid profile and apolipoprotein B. Lipid peroxidation was found to be significantly suppressed in extract-fed diabetic rats. The significant reduction in serum levels of oxidized low-density lipoprotein, soluble vascular cell adhesion molecule and plasma fibrinogen with a concomitant elevation in serum nitric oxide was observed in diabetic rats following treatment with extract. Histopathological examination of heart myocardium of extract-treated diabetic rats revealed reversal of fatty change toward normal. These results suggest that C. auriculata aqueous leaf extract exhibits anti-atherosclerotic role in the diabetic state and it indicates toward the notion that extract may help to prevent the progression of cardiovascular diseases.

Antiatherosclerotic Potential of Active Principle Isolated from Eugenia jambolana in Streptozotocin-Induced Diabetic Rats.

The aim of the present study was to investigate the antiatherosclerotic effect of active principle (FIIc) isolated from aqueous fruit pulp extract of Eugenia jambolana. Crude aqueous extract of E. jambolana was subjected to purification using chromatographic techniques which yielded purified active compound (FIIc). Purity of FIIc was tested by HPLC. Phytochemical investigation of FIIc by NMR, IR, and UV spectra showed that the purified compound is α-hydroxy succinamic acid. The streptozotocin- (STZ-) induced diabetic rats were fed atherosclerotic (Ath) diet containing 1.5 mL olive oil containing 8 mg (3, 20,000 IU) vitamin D(2) and 40 mg cholesterol for 5 consecutive days. The STZ-induced diabetic rats receiving Ath diet were orally administered FIIc at doses of 10, 15, and 20 mg/kg, and results were compared with reference drug, that is, glibenclamide (600 µg/mg) and healthy control. 30-day treatment with FIIc resulted in significant (P < .001) improvement in blood glucose, serum lipid profile, apolipoproteins (Apo A(1) and apoB(100)), and endothelial dysfunction parameters. Histomorphological studies also confirmed biochemical findings. Our results showed that FIIc has protective effect on hyperglycemia-induced atherosclerosis.

Protective effect of Morus rubra L. leaf extract on diet-induced atherosclerosis in diabetic rats.

The antiatherosclerotic effect of aqueous leaves extract of Morus rubra was studied in streptozotocin-induced diabetic rats fed with atherosclerotic (Ath) diet [1.5 ml olive oil containing 8 mg (3, 20,000 IU) vitamin D2 and 40 mg cholesterol] for 5 consecutive days. A short-term toxicity assessment was also conducted in healthy rats to examine toxic effects of the extract. Oral administration of extract to diabetic rats (100, 200 and 400 mg/kg body weight per day for a period of 30 days) produced significant (p<0.001) fall in fasting blood glucose (FBG) in a dose-dependent manner. Treatment with the extract (400 mg/kg) showed significant (p<0.001) improvement in body weight and serum lipid profile i.e., total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol and VLDL-cholesterol, when compared with diabetic control. Endothelial dysfunction parameters (sVCAM-1, Fibrinogen, total NO levels and oxidized LDL), apolipoprotein A and apolipoprotein B were significantly (p<0.001) reversed to near normal, following treatment with the extract. Thus, our study shows that aqueous leaf extract of Morus rubra (400 mg/kg) significantly improves the homeostasis of glucose and fat and possesses significant anti-atherosclerotic activity.

Attenuation of renal dysfunction by anti-hyperglycemic compound isolated from fruit pulp of Eugenia jambolana in streptozotocin-induced diabetic rats.

The renal protective effect of an active principle isolated from the aqueous extract of fruit pulp of Eugenia jambolana was investigated in streptozotocin (45 mg/kg body weight)-induced severely diabetic rats (FBG > or = 300 mg/dl). For isolation of active principle, crude aqueous extract of E. jambolana fruit pulp was subjected to purification by ion-exchange column chromatography, which yielded a partially purified compound (FII), which on further purification by rechromatography gave a purified active compound (FIIc). Purity of FIIc was confirmed by high pressure liquid chromatography. Detailed UV, NMR, IR spectra suggested that FIIc is a small aliphatic organic compound having molecular formula C4H7O4N. Oral administration of FIIc to diabetic rats (10, 15 and 20 mg/kg body weight per day for a period of 60 days) produced significant (P<0.001) fall in fasting blood glucose (FBG) in a dose-dependent manner. Treatment with FIIc (15 mg/kg body wt.) showed significant (P<0.001) improvement in body weight, blood urea, plasma creatinine levels, urinary volume, urinary sugar and microalbuminuria. Renal hypertrophy, assessed as the ratio of the weight of the two kidneys to total body weight was also significantly (P<0.05) improved after treatment with FIIc. The above results suggest that FIIc possesses significant nephroprotective activity.