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Background. Subarachnoid hemorrhage (SAH) is an important subcategory of stroke due to its high mortality rate as well as severe complications such as neurological deficit. It has been suggested that cerebral inflammation is a major factor in advanced brain injury after SAH. Microglia and astrocytes are known supporting cells in the development and maintenance of inflammation in central nervous system. However, the role of microglia and astrocytes in the development of inflammation and neuronal cell apoptosis during the early phase after SAH has not been thoroughly investigated. Materials and Methods. Sprague-Dawley rats were divided into 4 groups (n = 6 /group): sham group, animals subjected to SAH without treatment, SAH animals pretreated with the microglia inhibitor minocycline (50 mg/kg, ip), and SAH animals pretreated with the astrocyte inhibitor fluorocitrate (50 mg/kg, ip). SAH was induced by injecting autologous blood (1 ml/kg) into the cistern magna on day 0. Pretreatment with minocycline or fluorocitrate was given three days prior to the induction of SAH. Rats were sacrificed 6 hr after SAH, and their cerebral spinal fluids were used to measure protein levels of neuroinflammatory cytokines IL-1β, IL-6, and TNF-α by ELISA. In addition, the cerebral cortex was utilized to determine the levels of caspase-3 by western blot and to evaluate neuronal cell apoptosis by immunohistochemistry staining and detect microglia and astrocyte by immunofluorescence staining for Iba-1 and GFAP. In this study, all SAH animals were given an injection of autologous blood and SAH rats treated with minocycline or fluorocitrate received ip injections on day 1, 2, and 3 before inducing SAH. Neurological outcome was assessed by ambulation and placing/stepping reflex responses on day 7. Results. Immunofluorescence staining showed that SAH induced proliferation of microglia and astrocyte and minocycline inhibited the proliferation of both microglia and astrocyte. However, fluorocitrate inhibited only the proliferation of astrocyte. ELISA analysis showed that SAH upregulated TNF-α and IL-1β, but not IL-6 at 6 hr after SAH. Minocycline, but not fluorocitrate, attenuated the upregulation of TNF-α and IL-1β. Western blot analysis and immunohistochemistry staining showed that SAH induced neuronal cell apoptosis. Pretreatment with minocycline, but not fluorocitrate, decreased SAH-induced neuronal death and cerebral vasospasm. Furthermore, significant improvements in neurobehavioral outcome were seen in the minocycline treatment group, but not in animals treated with fluorocitrate. Conclusions. Microglia may play an important role to regulate neuronal cell apoptosis and cerebral vasospasm through inhibiting inflammation at an early phase after SAH in the rat. [ABSTRACT FROM AUTHOR]

Aims Many immunohistochemical markers have been studied for differentiating papillary lesions. However, their differentiating power has not been evaluated comprehensively. Additionally, assessment of some markers will require the difficult task of identifying different cell types. In the current study, we aimed to devise a simple papillary panel which can aid in diagnosis irrespective of architectural pattern and cell type differentiation. Methods and results Immunohistochemical analysis of papillary lesions using myoepithelial markers [p63 and smooth muscle actin (SMA)], high molecular weight cytokeratins (HMWCKs: CK5, CK5/6, CK14 and 34βE12), hormone receptors (ER and PR) and neuroendocrine markers (chromogranin and synaptophysin) was performed. Among them, neuroendocrine markers showed high specificity but low sensitivity. HMWCK specificity was better than that for myoepithelial markers. Homogeneous staining pattern for hormonal receptors rather than their percentage positivity was more effective in identifying malignant lesions. Negative staining for two or more of HMWCKs, namely CK5/6, CK14 and 34βE12, achieved the best overall specificity and sensitivity of 87.8% and 94.1%, respectively, irrespective of the architecture. Their discriminatory power was validated with an independent cohort of core needle biopsies. Conclusions A marker panel with HMWCKs could be used in differentiating papillary lesions irrespective of architectural pattern or cell type differentiation. [ABSTRACT FROM AUTHOR]

Aims SOX2 is a key regulatory gene in embryonic stem cells. Although it has been implicated in cancer progression, its role in breast carcinoma is poorly understood. Materials and methods Fifty-seven ductal carcinomas in situ ( DCIS), 552 invasive breast carcinomas and 107 corresponding metastatic lymph nodes were evaluated immunohistochemically for the expression of SOX2. Its correlation with clinicopathological features, other biomarker profiles and patients' outcomes were analysed. Results SOX2 was detected in 19.0% (105 of 552) of invasive breast carcinomas and 12.3% (seven of 57) of DCIS. Expression correlated with larger tumour size ( P = 0.005) and higher grade ( P = 0.002). It was associated negatively with ER ( P = 0.015) and PR ( P = 0.046) expression, but positively with Ki67 index ( P = 0.013). Interestingly, it was also associated with neuroendocrine marker expression (synpatophysin and chromogranin/synaptophysin, P = 0.048 and 0.028, respectively). Expression appeared to be independent from that of common stem cell markers, namely CD44, CD24 and aldehyde dehydrogenase 1 ( ALDH1). Furthermore, a higher rate of expression was observed in metastatic lymph nodes than in the corresponding primary tumours ( P = 0.034). High SOX2 expression was correlated with poor disease-free survival (log-rank=9.489, P = 0.012) and was an independent prognostic factor ( HR=2.918, P = 0.015) in patients with high nodal stages. Conclusions In summary, SOX2 expression was related to adverse breast carcinoma profile and poor outcome in selected patient groups. [ABSTRACT FROM AUTHOR]

Immunohistochemical analysis of gross cystic disease fluid protein-15 (GCDFP-15) and mammaglobin (MGB) is frequently used in routine practice for assessment of metastases or regional recurrences of breast origin. Breast cancer is highly heterogeneous. Expression of these 2 markers in various breast cancer subtypes has not been well studied. In addition, the usefulness of these two markers in combination in detecting breast origin has not been explored. In this study, a large cohort of breast cancers was evaluated for GCDFP-15 and MGB expression, both individually and combined. Their expression was correlated with cancer subtypes, other biomarkers and clinicopathologic parameters. A higher sensitivity for MGB (42.3%) than GCDFP-15 (31.6%) in detecting cancers of breast origin was observed. Combining both increased the sensitivity further, both for primary tumor (53.0%) and for nodal metastases (69.0%). GCDFP-15 was associated significantly with a breast cancer profile of good prognosis tumors, including lower grade (P < .001), pN (P = .029) and Ki-67 (P < .001) as well as negative basal markers expression (P = .043, .009, and .049 for c-Kit, CK5/6 and epidermal growth factor receptor, respectively) and, thus, may not be sensitive for detection of poor prognosis tumors. MGB has the highest expression in HER2-overexpressing cancers (56.6%), and may be a potentially useful marker for this subtype. Nonetheless, both markers showed low expression in the basal like (BLBC) subtype (11.9% and 21.4% for GCDFP-15 and MGB respectively), therefore, the detection of BLBC remains problematic. Negative results need to be interpreted with caution, and correlation with other clinical findings may be required to exclude the possibility of metastatic BLBC. [ABSTRACT FROM AUTHOR]