Potential significance of podoplanin immunohistochemical expression in papillary thyroid carcinoma.

Podoplanin (PDPN) is a lymphatic endothelial marker expressed by a range of human malignancies in which it has been shown to contribute to tumor progression and metastasis. However, there is a lack of the studies, examining the function of PDPN in thyroid cancer. The current study was performed to explore the possible diagnostic value of PDPN expression in papillary thyroid cancer (PTC) and to evaluate the marker's potential for prediction of regional lymph node metastasis. Lymphatic vascular density (LVD) and the stromal/cancer-associated fibroblasts (CAFs), labeled by PDPN, were examined in PTC compared to the other thyroid lesions. The current study included 50 cases of PTC and 50 cases of non-PTC thyroid lesions. Immunohistochemical staining was performed using monoclonal PDPN antibodies. Podoplanin expression was scored as positive and negative. Podoplanin expression was found in 36% of PTC cases, but it was not found in benign, low risk (borderline), or malignant lesions other than PTC. Furthermore, lymph node metastasis was significantly correlated with PDPN expression, LVD and CAFs (p-values < 0.00001, < 0.001 and 0.0002 respectively). These findings support the diagnostic utility of PDPN expression in PTC and its predictive value for LN metastasis.

Emerging Role of Nestin as an Angiogenesis and Cancer Stem Cell Marker in Epithelial Ovarian Cancer: Immunohistochemical Study.

Ovarian cancer is the most fatal gynecologic malignancy and the existing second-line treatments have not been confirmed to be effective. Cancer stem cells research has a leading role to explore promising therapeutic applications. Nestin was postulated to reflect cancer stem cell properties in various tumors, correlating with poor prognosis. Furthermore, nestin is proposed as a reliable neovascularization marker. This study aimed to elucidate the status of nestin expression in various epithelial ovarian cancers (EOCs), its neoangiogenic properties, and investigate its potential association with clinicopathologic parameters. A total of 80 primary EOCs (37 serous, 20 Mucinous, 13 endometrioid, and 10 clear cell carcinomas) were immunohistochemically stained with nestin. Staining intensity and automated microvascular density (MVD) were assessed. Positive nestin expression was defined in ≈47.5% of all EOC; more commonly in ≈60% of the serous tumors. It was noticeably expressed in tumor spheroids. Nestin expression significantly correlated with overall tumor grade, lymph node, distant metastasis, and stage. Nestin neoangiogenesis was detectable in all cases (average=60.1). The nestin expression in tumor cells significantly correlated with Nestin/MVD. The average Nestin/MVD was significantly an independent predictor of high tumor stage. As a stem cell marker, nestin is expressed in cells of EOC including those growing as spherules and correlated with poor prognosis. Thus, nestin may be a novel therapeutic target for tumor angiogenesis and a combination therapy that includes nestin-targeting agents may be an effective therapeutic approach. In addition, detection of Nestin/stem cells and Nestin/MVD can be used as predictors of disease.

Aliskiren attenuates bleomycin-induced pulmonary fibrosis in rats: focus on oxidative stress, advanced glycation end products, and matrix metalloproteinase-9.

Pulmonary fibrosis is a progressive lung disorder with high mortality rate and limited successful treatment. This study was designed to assess the potential anti-oxidant and anti-fibrotic effects of aliskiren (Alsk) during bleomycin (BLM)-induced pulmonary fibrosis. Male Wistar rats were used as control untreated or treated with the following: a single dose of 2.5 mg/kg of BLM endotracheally and BLM and Alsk (either low dose 30 mg/kg/day or high dose 60 mg/kg/day), and another group was given Alsk 60 mg/kg/day alone. Alsk was given by gavage. Alsk anti-oxidant and anti-fibrotic effects were assessed. BLM significantly increased relative lung weight and the levels of lactate dehydrogenase and total and differential leucocytic count in bronchoalveolar lavage that was significantly ameliorated by high-dose Alsk treatment. As markers of oxidative stress, BLM caused a significant increase in the levels of lipid peroxides and nitric oxide accompanied with a significant decrease of superoxide dismutase and glutathione transferase enzymes. High-dose Alsk treatment restored these markers toward normal values. Alsk counteracted the overexpression of advanced glycation end products, matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinases-1 in lung tissue induced by BLM. Fibrosis assessed by measuring hydroxyproline content, which markedly increased in the BLM group, was also significantly reduced by Alsk. These were confirmed by histopathological and immunohistochemical examination which revealed that Alsk attenuates signs of pulmonary fibrosis and decreased the overexpressed MMP-9 and transforming growth factor ß1. Collectively, these findings indicate that Alsk has a potential anti-fibrotic effect beside its anti-oxidant activity.

Revisiting the Role of the Myofibroblast in Socket Surgery: An Immunohistochemical Study.

PURPOSE: To determine the impact of a single injection of various anti-inflammatory, antimitotic, and antiangiogenic agents on the cell count of myofibroblasts in an eviscerated socket. METHODS: One eye from 15 skeletally mature New Zealand white rabbits was eviscerated, and the rabbits were divided into 5 groups. Each group of 3 rabbits received a 0.1 ml subconjunctival injection of a different agent. Group I received bevacizumab 25 mg/ml, group II received triamcinolone 40 mg/ml, group III received 5-fluorouracil 50 mg/ml, group IV received mitomycin-C 0.4 mg/ml, while group V was the control group and received no injections. The animals were euthanized 19 days after evisceration and conjunctival samples were submitted for histopathological examination. Monoclonal α-smooth muscle actin antibody was applied, and the mean of 5 readings of the number of myofibroblasts was recorded in each slide. RESULTS: The mean count of myofibroblasts was highest for the control group and all groups achieved a statistically significant reduction in myofibroblast count compared with the control group. Sorting the means showed that Group IV (mitomycin-C) achieved the lowest mean value (p = 0.000006) followed by triamcinolone (p = 0.00048), while group I (bevacizumab) achieved the least reduction in myofibroblast count (p = 0.00148). CONCLUSION: Until newer antimyofibroblast medications and antibodies are commercially available, a single injection of mitomycin-C or triamcinolone during surgery achieves the highest mean reduction of myofibroblast count.

Combined use of COX-1 and VEGF immunohistochemistry refines the histopathologic prognosis of renal cell carcinoma.

The course of RCC is asymptomatic, resulting in 25-30% of patients presenting with metastatic disease at time of diagnosis. The development of novel agents targeting angiogenesis and signal transduction pathways has improved patient outcomes. Role of cyclooxygenase in cancer development has been the subject of close scrutiny. COX-1 has been recognized to be involved in regulation of angiogenesis. To date, no immunohistochemical studies have been performed to assess the possible association between COX-1 and VEGF in RCC. This study is designed to evaluate the relationship between these two proteins in RCC. Also, the relationship between their combined immunohistochemical expression and different clinicopathological prognostic parameters in RCC is investigated. Immunohistochemical expression of COX-1 and VEGF was evaluated retrospectively on 64 cases of primary RCC including: 45 clear cell carcinoma, 12 papillary carcinoma and 7 of chromophope carcinoma. High COX-1 expression was detected in 62.5% of RCCs with a significant association with tumor grade (P=0.028), and highly significant relationship with tumor size and stage (P=0.001). There was a highly significant relationship between the VEGF score and tumor size (P=0.001), and stage (P=0.006). There was a positive correlation between COX-1 and VEGF expression score (P=0.001). Combined expression of both markers predicts high stage tumors (stage III/IV). Immunohistochemical expression of COX-1 and VEGF is associated with poor prognostic parameters in RCC. Their combined expression has a beneficial role in prediction of high stage tumors (III/IV).

Relationship between osteopontin and ß-catenin immunohistochemical expression and prognostic parameters of colorectal carcinoma.

Previous studies on the prognostic value of osteopontin (OPN) and ß-catenin in colorectal carcinoma (CRC) revealed conflicting results. To date, only two immunohistochemical studies investigated their association in CRC with discrepant results. Moreover, the relevance of their co-expression to clinicopathological parameters was not previously reported. This study was designed to investigate the relationship between these markers and prognostic parameters in CRC and study further the relationship between them. Immunohistochemical expression of OPN and ß-catenin was evaluated in 72 CRCs. Cytoplasmic OPN was detected in 45.83% of CRCs while normal mucosa was immunonegative. Strong continuous membranous ß-catenin was present in normal mucosa. However, abnormal membranous, nuclear and cytoplasmic expressions were observed in 36.11%, 31.94% and 52.78% of CRCs, respectively. A highly significant relationship was detected between each of OPN and nuclear ß-catenin expression and lymph node metastasis (P = 0.0001 and 0.004 respectively), depth of invasion (P = 0.001 and 0.004 respectively), TNM stages (P = 0.0001 and 0.001 respectively) and Dukes' stages (P = 0.0001 and 0.004 respectively). A significant association was found between OPN and distant metastases. A strong agreement was observed between OPN and nuclear ß-catenin (kappa = 0.656). A highly significant relationship was found between their co-expression and poor prognostic parameters. OPN overexpression and nuclear ß-catenin expression appeared to be associated with unfavorable prognostic factors in CRC. A direct relationship was observed between them. Further understanding their role in colorectal carcinogenesis as well as targeting the interaction between them might be effective in the future development of therapeutic agents for CRC patients.

DJ-1 and androgen receptor immunohistochemical expression in prostatic carcinoma: a possible role in carcinogenesis.

BACKGROUND AND AIM: Androgen plays a fundamental role in the growth and differentiation of prostate. Androgen receptor (AR) expression may represent a potential marker of prognosis in prostate cancer. However, there have been variable results regarding its ability to predict clinical progression. Despite the oncogenic properties of DJ-1, its significance in prostate cancer development and progression is not well understood. This research shed some light on the possible role of immunohistochemical expression of DJ-1 in clinically localized prostatic carcinoma in relation to the established role of AR and other clinicopathologic parameters. MATERIALS AND METHODS: The immunohistochemical expression of AR and DJ-1 was evaluated in 129 samples including benign hyperplasia (n = 60) and prostatic carcinoma (n = 69). RESULTS: The mean value of AR immunostaining was significantly higher in prostatic carcinomas than in benign hyperplasia (P = 0.001). A significant inverse correlation was found between AR immunostaining and the grade of prostatic carcinomas. A significantly higher median DJ-1 score was found in prostatic carcinoma than in benign hyperplasia (P = 0.0001). There was a significant direct correlation between AR and DJ-1 score (P = 0.0001). AR is more sensitive in predicting prostatic carcinoma than DJ-1 but DJ-1 is more specific than AR. CONCLUSION: AR nuclear expression was consistently present in benign and adenocarcinoma epithelium. But, there may be limited clinical use for AR expression in localized carcinoma due to its constant heterogeneity. DJ-1 with its oncogenic properties, specificity for prostatic carcinoma and homogenous expression gives an ideal complementary role to AR in the detection and treatment of prostatic carcinomas.

Insights into the prognostic value of DJ-1 and MIB-1 in astrocytic tumors.

BACKGROUND: The histological grade is the gold standard for the evaluation of prognosis of astrocytic tumors. Nevertheless, morphologic criteria are not always accurate prognostic indicators. AIM: The research investigates the expression of MIB-1 and DJ-1 in different grades of astrocytomas and evaluates the possible prognostic role of DJ-1 in these tumors in relation to other prognostic parameters including the MIB-1 labeling index. MATERIALS AND METHODS: Immunohistochemical expression of MIB-1 and DJ-1 was evaluated in 111 samples of astrocytic tumors comprising 28 diffuse astrocytomas, 38 anaplastic astrocytomas and 45 glioblastomas. The univariate survival analysis was done using the Kaplan-Meier method and the multivariate survival analysis was done using Cox proportional hazard model. RESULTS: The statistical analysis revealed a significant correlation between each of DJ-1 and MIB-1 and the histological grade of astrocytomas. The univariate analysis showed that high grade, high DJ-1 score and MIB-1 labeling index ≥ 10.1 were associated with poor survival. Multivariate analysis for all the studied astrocytomas proved the independent prognostic significance of the histological grade and DJ-1 score. Meanwhile, the multivariate analysis for each grade emphasized that DJ-1 was the only independent prognostic indicator in high-grade astrocytomas. CONCLUSION: This study emphasized the effectiveness of high DJ-1 expression in predicting poor survival of astrocytoma patients, when compared to MIB-1. DJ-1 could be particularly important in cases with discrepancies between the morphologic criteria and clinical parameters. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1070116023943146.

Skin tags: a link between lesional mast cell count/tryptase expression and obesity and dyslipidemia.

BACKGROUND: The etiology of skin tags (STs) is not fully understood. A relation to diabetes mellitus and obesity was suggested. Few studies of possible mast cells (MCs) involvement were reported. Tyrptase is a mast cell mediator and a potent fibroblast growth factor. It may provide a molecular link between mast cell activation and fibrosis. AIMS: The aim was to assess clinical and laboratory findings in patients with STs, and the possible link between obesity, dyslipidemia, and lesional MC count/tryptase expression. MATERIALS AND METHODS: A total of 20 patients with STs were subjected to clinical examination, estimation of body mass index (BMI), fasting blood glucose (FBG), postprandial blood glucose (PPBG), serum cholesterol and triglycerides, abdominal ultrasound for fatty liver assessment, in addition to study of MCs through staining for MC tryptase in two skin biopsies; lesional and nonlesional (control). RESULTS: All patients showed abnormally high BMI and hypertriglyceridemia, with abnormal sonographic pattern in 15 patients (75%). STs number positively correlated with the age of patients. STs showed significantly higher MC counts and tryptase expression, compared with control skin (P < 0.001), with no correlation of the STs number or MC count with BMI, FBG, PPBG or serum cholesterol. Obese patients showed a significantly higher MC count than overweight and there was a positive correlation between MC count and serum triglycerides. Axilla and under breast STs showed a higher MC count compared with other sites. CONCLUSIONS: STs seem to be related to obesity and hypertriglyceridemia. MCs with their tryptase are possibly involved in pathogenesis of STs. MC count is related to the associated factors; obesity and serum triglycerides. MC tryptase expression is a reliable method for accurate tissue MC counting.

N-terminal pro-brain natriuretic peptide in systemic sclerosis: new insights.

OBJECTIVE: To identify the relation between serum levels of N-terminal pro-brain natriuretic peptide (NT-pro BNP) and severity of systemic sclerosis (SSc). SUBJECT AND METHODS: 25 SSc patients and 25 age- and sex-matched healthy controls were included. Assay of serum NT-pro BNP was done for all patients and controls. Patients were subjected to modified Rodnan skin score (mRss), echocardiography, pulmonary function tests and skin biopsies for histopathological skin thickness score assessment. There was a significant increase in the mean values of serum levels of NT- pro BNP in SSc patients compared to controls (t=11, p<0.001). RESULTS: There was a significant positive correlation between serum levels of NT-pro BNP in SSc patients and mRss, systolic pulmonary artery pressure (sPAP) and histopathological skin thickness score (r=0.93, r=0.92, r=0.92, p<0.001 respectively). There was a significant increase in the mean value of serum levels of NT-pro BNP and sPAP in SSc patients with restrictive pulmonary affection compared to those with normal respiratory function tests (p<0.001). CONCLUSION: NT-pro BNP may be a useful biological marker for assessing the severity of SSc as it has a role in detecting the extent of skin fibrosis, the severity of PAH and the degree of restricted pulmonary involvement in SSc patients.

Differential expression of cyclin D1 in human pituitary tumors: relation to MIB-1 and p27/Kip1 labeling indices.

BACKGROUND: Pituitary tumors are a common form of endocrine neoplasia. However few studies assessed the expression of the principal cyclin regulating checkpoint exit, cyclin D1. Cyclin D1 expression in pituitary tumors and its possible relation to MIB-1 and p27/Kip1 labeling indices (LIs) was explored. DESIGN: We studied a total of 199 pituitaries, including normal pituitaries (n=7), pituitary adenomas (n=187), and pituitary carcinoma (n=5). All tissues were tested as cores of archived tissue microarrays that were immunostained for cyclin D1, MIB-1 and p27 using a standard technique. Tissue cores were subjected to automated analysis to evaluate the staining LIs. RESULTS: No cyclin D1 positive cells in the normal anterior pituitary gland was found. Sparse nuclear staining was noted in pituitary tumors. Higher expression of cyclin D1 was noted in pituitary carcinomas compared to adenomas (p<0.001), in non-functioning adenomas compared to functioning ones (p<0.001) in macroadenomas versus microadenomas (p=0.017) and in recurrent non recurrent adenomas (p<0.001). Cyclin D1 LI and MIB-1 LI were related among adenomas (p<0.001) and carcinomas (p=0.041). p27 LI was neither related to pituitary adenoma recurrence nor invasion. CONCLUSIONS: Expression of cyclin D1 in pituitary tumors is related to cell proliferation, recurrence, and metastatic potential. Nuclear cyclin D1 expression is a good marker of aggressive behavior in pituitary tumors.

Keratinocyte and lymphocyte apoptosis: relation to disease outcome in systemic lupus erythematosus patients with and without cutaneous manifestations.

Our aim was to assess the relationship of keratinocyte and lymphocyte apoptosis and macrophage function to disease outcome in systemic lupus erythematosus patients with and without cutaneous manifestations. 50 systemic lupus erythematosus patients [25 with cutaneous manifestations (group I), 25 without cutaneous manifestations (group II)] and 20 normal controls (group III) were studied. Assessments of disease activity, peripheral lymphocyte apoptosis, macrophage function and apoptotic cells in skin and renal biopsies were carried out. The mean systemic lupus erythematosus disease activity index score was significantly higher in group I than II (18.6 +/- 6, 8.8 +/- 2.7 respectively, p < 0.001). The mean percentage of peripheral apoptotic lymphocytes was significantly higher in group I than groups II, III (55.3 +/- 21.4, 25.6 +/- 8.7 & 19.4 +/- 3.2 respectively, P < 0.001), so was serum neopterin level (27.5 +/- 7.3, 14.9 +/- 2.7, 9.4 +/- 1.1 respectively, p < 0.001), and the mean number of protein53 positive apoptotic keratinocytes in skin (20.6 +/- 5.4, 1.6 +/- 0.5, 1.7 +/- 0.4 respectively, p < 0.001). A higher percentage of class IV, V glomerulonephritis was found in group I (47%, 26%, respectively) compared to group II (11% both) (p < 0.001). The mean number of protein53 positive apoptotic skin keratinocytes showed a significant positive correlation to disease activity, percentage of peripheral apoptotic lymphocytes and serum neopterin (P < 0.001). In conclusion, an accumulation of apoptotic keratinocytes and lymphocytes in systemic lupus erythematosus with cutaneous manifestations is associated with a worse disease outcome.

Impact of experimental duel infections with Schistosoma mansoni and Echinoccocus granulosus on hepatic histopathology.

Experimental duel infection with S. mansoni and E. granulosus was induced in mice to determine their effect on serum nitric oxide (NO) level and accordingly on the sequences of histopathological lesions affecting the liver. The results showed that serum NO level was significantly increased (p<0.05) in mice infected with both parasites (GI) in comparison to either S. mansoni (GIV) or E. granulosus (GV). The NO elevation on hepatic pathological lesions of both diseases showed a marked reduction of granuloma size with absence of concentric fibrosis in GI as early as 4 weeks of concomitant infection as compared to GIV. In spite of the significant increase of NO level when E. granulosus infection induced in late stages of schistosomisais (GsII & III), yet granuloma size was not suppressed. Also, there was absence or death of hydatid cyst in mice (GI) compared to E. granulosus (GV). So, the duel infection with the two parasites affected serum NO level and hepatic histopathology, by ameliorative or deteriorative effects, according to duration of infection with either.