Human endoplasmic reticulum oxidoreductin 1-a (hERO1-a) is an oxidizing enzyme that exists in the endoplasmic reticulum and its expression is augmented under hypoxia. It regulates a redox state of various kinds of protein through reoxidation of "client" protein disulfide isomerase. Interestingly, although the expression of hERO1-a in normal tissues was comparatively limited, various types of cancer cells expressed it in large amounts. Therefore, we examined the role of ERO1-a in tumor growth using murine breast cancer line 4T1 and found that knockdown of murine ERO1-a inhibited in vivo tumor growth and decreased lung metastasis compared with wild-type 4T1. Moreover, we investigated the relationship between expression of hERO1-a and prognosis in breast cancer patients. Seventy-one patients with breast cancer who underwent surgery between 2005 and 2006 in Sapporo Medical University Hospital (Sapporo, Japan) were analyzed in this study. Significant differences were found between the hERO1-a-positive group (n = 33) and hERO1-a-negative group (n = 38) in nuclear grade (P < 0.001) and intrinsic subtype (P = 0.021) in univariate analysis. More importantly, in multivariate analysis of disease-free survival by Cox regression, expression of hERO1-a was the only independent prognosis factor (P = 0.035). Finally, in univariate survival analysis, patients positive for hERO1-a had significantly shorter disease-free survival and overall survival than those patients negative for hERO1-a. These findings indicate that the expression of hERO1-a in cancer cells is associated with poorer prognosis and thus can be a prognostic factor for patients with breast cancer. (Cancer Sci 2013;

Our results confirm the need to test for HRPT2 in FIHP families, especially those with parathyroid carcinomas, atypical adenomas or adenomas with cystic change.

Multiple system atrophy (MSA) is a neurodegenerative disease caused by an accumulation of ␣-synuclein (␣-syn) in oligodendrocytes. Little is known about the cellular mechanisms by which ␣-syn accumulation causes neuronal degeneration in MSA. Our previous research, however, revealed that in a mouse model of MSA, oligodendrocytic inclusions of ␣-syn induced neuronal accumulation of ␣-syn, as well as progressive neuronal degeneration. Here we identify the mechanisms that underlie neuronal accumulation of ␣-syn in a mouse MSA model. We found that the ␣-syn protein binds to ␤-III tubulin in microtubules to form an insoluble complex. The insoluble ␣-syn complex progressively accumulates in neurons and leads to neuronal dysfunction. Furthermore, we demonstrated that the neuronal accumulation of insoluble ␣-syn is suppressed by treatment with a microtubule depolymerizing agent. The underlying pathological process appeared to also be inhibited by this treatment , providing promise for future therapeutic approaches.

The separation and identification of pigments, chlorophylls, and carotenoids of seven teas and fresh leaf of tea (Camellia sinensis) by high-performance liquid chromatography (HPLC) are described. HPLC was carried out using a Symmetry C(8) column with a photodiode array detector. Pigments were eluted with a binary gradient of aqueous pyridine solution at a flow rate of 1.0 mL/min at 25 degrees C. HPLC analyses achieved the separation of more than 100 pigment peaks, and 79 pigment species, 41 chlorophylls, and 38 carotenoids were detected. The presence of degraded chlorophylls was a common feature, and the number and the variety of pigments differed with tea species. Generally, the numbers of chlorophyll species tended to increase with processing steps, while carotenoid species were decreased, especially by heating. Particularly in green teas, a change of carotenoid structure, conversion of violaxanthin to auroxanthin, occurred. In hot water extracts of teas, both chlorophylls and carotenoids were also detected, but the concentration of chlorophylls was less than 2% as compared with acetone extracts. The pigment compositions were compared between tea species, and they are discussed in terms of the differences in their manufacturing processes.

Enzymatic removal of the methoxycarbonyl group of pheophorbide (Pheid) a in chlorophyll degradation was investigated in cotyledons of radish (Raphanus sativus). The enzyme pheophorbidase (PPD) catalyzes the conversion of Pheid a to a precursor of pyropheophorbide (PyroPheid), C-13 2 -carboxylPyroPheid a, by demethylation, and then the precursor is decarboxylated nonenzymatically to yield PyroPheid a. PPD activity sharply increased with the progression of senescence in radish, suggesting de novo synthesis of PPD. The enzyme activity was separated into two peaks in anion-exchange and hydrophobic chromatography; the terms type 1 and type 2 were applied according to the order of elution of these enzymes in anion-exchange chromatography. PPD types 1 and 2 were purified 9,999-and 6,476-fold, with a yield of 0.703% and 2.73%, respectively. Among 12 substrates tested, both enzymes were extremely specific for Pheids of the dihydroporphyrin and tetrahydroporphyrin types, indicating that they are responsible for the formation of these PyroPheids. Both PPDs had molecular masses of 113,000 kD on gel filtration and showed three bands of 16.8, 15.9, and 11.8 kD by SDS-PAGE. The partial N-terminal amino acid sequences for these bands of PPD (type 2) were determined. Based on their N-terminal amino acid sequences, a full-length cDNA of PPD was cloned. The molecular structure of PPD, particularly the molecular mass and subunit structure, is discussed in relation to the results of SDS-PAGE.