Binding of chemicals to PDI and their effects on the catalytic activity and physiological functions of PDI

関西学院大

ニワトリ卵巣での卵成熟に伴う卵黄膜内層タンパク質の変化

To clarify the formation of the vitelline membrane, we investigated changes in the proteins constituting the inner layer of the vitelline membrane during the ovum maturation in the hen ovary.The vitelline membranes were isolated from yellow yolk follicles at the different growth stages, identified as F_1 to F_8,and their protein compositions were examined by SDS-PAGE in both the presence and the absence of reducing agent. One of the major glycoproteins found in the matured ovum, GP-I, appears to exist as monomer from the early stage of the yolk deposition (F_1). The other glycoprotein, GP-II, appears as protomer of its precursor with higher molecular weight at slightly later stage (F_3) and appears to be converted into the homodimeric form at later stages (F_7～F_8). Staining the gel for the carbohydrate moiety supports this notion. The GP-I fraction prepared from the vitelline membrane of the completed eggs was separated into two components, GP-Ia and GP- Ib, based on the pH dependence in their solubility. They have similar molecular weights but different elution volumes in cation-exchange chromatography. A similar but different heterogeneity in the GP- I fraction prepared from the follicle vitelline membrane was identified.These results indicate that proteins constituting the vitelline membrane, at least GP-I and GP-II, are processed to a great extent during the ovum maturation as well as after the ovulation

組織因子惹起-トロンボモジュリン添加凝固波形解析を用いたプロテインC経路異常を伴う血栓性素因のスクリーニング法

Objectives: Absolute or relative protein (P)C pathway abnormalities (PC deficiency, PS deficiency, antiphospholipid syndrome (APS), factor (F)V-abnormality, and high FVIII level) cause thrombophilia. Although screening assays for these thrombophilias are available, one utilizing clot waveform analysis (CWA) remains unknown. We aimed to establish a CWA-based screening assay to distinguish PC pathway abnormality-related thrombophilia. Methods: Samples were reacted with tissue factor (TF)/phospholipids and recombinant thrombomodulin (rTM; optimal 20 nM), followed by CWA measurement. The peak ratio (with/without rTM) of the first derivative curve of clot waveform was calculated. Results: The peak ratio in healthy plasmas (n = 35) was 0.36 ± 0.13; hence, the cutoff value was set to 0.49. The peak ratios in plasmas with PC deficiency, PS deficiency, high-FVIII (spiked 300 IU/dl), and APS were higher than the cutoff values (0.79/0.97/0.50/0.93, respectively). PC-deficient plasma or PS-deficient plasma mixed with normal plasma (25%/50%/75%/100% PC or PS level) showed dose-dependent decreases in the peak ratios (PC deficient: 0.85/0.64/0.44/0.28; PS deficient: 0.69/0.53/0.40/0.25), suggesting that the peak ratio at ≤50% of PC or PS level exceeded the cutoff value. The peak ratio in FV deficiency with FV ≤25% was higher than the cutoff value. FV-deficient plasma spiked with 40 IU/dl rFV-R506Q (FVLeiden ) or rFV-W1920R (FVNara ) showed >90% peak ratios. Conclusions: rTM-mediated TF-triggered CWA might be useful for screening PC pathway abnormality-related thrombophilia.博士（医学）・乙第1527号・令和4年9月28日© 2022 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.This is the peer reviewed version of the following article: [https://onlinelibrary.wiley.com/doi/10.1111/ejh.13777], which has been published in final form at [https://doi.org/10.1111/ejh.13777]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.発行元が定める登録猶予期間終了の後、本文を登録予定（2023.07

卵管における卵黄膜外層タンパク質の分泌部位の解析

To clarify the formation of the vitelline membrane outer layer we examined the proteins secreted from hen oviduct by using Western blotting analysis. Antibodies were elicited against vitelline membrane outer layer protein-I (VMO-I) and against the one of subunit (S 8) of ovomucim obtained from albumen. It was found that VMO-I was mostly secreted from the middle region of the infundibulum. On the contrary, S 8 was secreted from the lower region of the infundibulum over the lower region of the magnum. These findings indicated that 1) VMO-I is present in the infundibulum prior to the formation of the outer layer, 2) the chalaziferous layer is formed at the upper region in the magnum, 3) the albumen is formed at the middle region to the lower region in the magnum

Dysbindin Regulates the Transcriptional Level of Myristoylated Alanine-Rich Protein Kinase C Substrate via the Interaction with NF-YB in Mice Brain

BACKGROUND: An accumulating body of evidence suggests that Dtnbp1 (Dysbindin) is a key susceptibility gene for schizophrenia. Using the yeast-two-hybrid screening system, we examined the candidate proteins interacting with Dysbindin and revealed one of these candidates to be the transcription factor NF-YB. METHODS: We employed an immunoprecipitation (IP) assay to demonstrate the Dysbindin-NF-YB interaction. DNA chips were used to screen for altered expression of genes in cells in which Dysbindin or NF-YB was down regulated, while Chromatin IP and Reporter assays were used to confirm the involvement of these genes in transcription of Myristoylated alanine-rich protein kinase C substrate (MARCKS). The sdy mutant mice with a deletion in Dysbindin, which exhibit behavioral abnormalities, and wild-type DBA2J mice were used to investigate MARCKS expression. RESULTS: We revealed an interaction between Dysbindin and NF-YB. DNA chips showed that MARCKS expression was increased in both Dysbindin knockdown cells and NF-YB knockdown cells, and Chromatin IP revealed interaction of these proteins at the MARCKS promoter region. Reporter assay results suggested functional involvement of the interaction between Dysbindin and NF-YB in MARCKS transcription levels, via the CCAAT motif which is a NF-YB binding sequence. MARCKS expression was increased in sdy mutant mice when compared to wild-type mice. CONCLUSIONS: These findings suggest that abnormal expression of MARCKS via dysfunction of Dysbindin might cause impairment of neural transmission and abnormal synaptogenesis. Our results should provide new insights into the mechanisms of neuronal development and the pathogenesis of schizophrenia

ゲンパツセイ コウジョウセンガン ノ ネンレイ ニヨル リンショウ ビョウリガクテキ トクチョウ オヨビ ヨゴ ノ ヒカク ケントウ

Two-hundred thirty-six patients with primary thyroid cancer who received operation were divided into two groups by age, i.e.,５９patients of age ＜４５years（Early adulthood, EA）and１７７ patients of age≧４５years（advanced age, AA）. Clinicopathologic factors and disease-free survival （DFS）were compared between the two groups. There was no difference in clinicopathologic factors except for higher proportion of patients with poorly differentiated adenocarcinoma in the AA patients than in the EA patients（６．７％ vs. ０％, p＝０．０４１）. DFS was significantly longer in the AA patients than in the EA patients（disease-free rates at１０years after operation，９４．８％ vs. ７２．５％, p＝０．００３１）. Overall survival was not different between the two groups. The EA patients who showed shorten DFS were divided into two groups，１７ patients of age ＜３０ years （juvenile and young adult, JYA）and ４２ patients of age ≧３０ years, and DFS of each group was compared with that of the AA patients. Although disease-free survival rates at １０ years of the JYA patients were not different（９２．６％ vs.９４．８％, p＝０．１２５）, those of patients of age ≧３０years were significantly lower than those of the AA patients（７０．０％ vs. ９４．８％, p＝０．００２１）. These findings suggest that patients with primary thyroid cancer who are ≧３０ years old in the young adulthood should be observed carefully after operation for early detection of relapse

若年性乳腺線維腺腫

A １２-year-old, premenarchal girl with a rapidly growing breast lump was transferred to our hospital. Ultrasonography revealed a ７．９×７．７×３．３cm large well-circumscribed hypoechoic tumor in the medial region of her right breast. After ２ months of this first visit, the breast lump had enlarged to１０ cm in diameter and was consequently subjected to a wide excision. Size of the tumor was９．５×９．２×３．０cm. Histopathological diagnosis was fibroadenoma without any malignant component. One year after the operation, a new lump with a diameter of４cm was detected in the lateral region of her right breast that had a similar appearance of the previous tumor. The tumor was excised as earlier and diagnosed as fibroadenoma. We present an extremely rare case of a rapidly growing breast fibroadenoma in a premenarchal girl. To the best of our knowledge, there were only ２１ cases reported in Japan. Since the patient is concerned that another asynchronous fibroadenoma might occur, she needs a long-term and careful follow-up

キョウクウキョウカ ニ セツジョ シタ ジュウカクナイ イショセイ フクコウジョウセン センシュ

A woman in her ６０s who was diagnosed as having hypercalcemia and hyperparathyroidism was referred to our hospital. Serum calcium and intact parathyroid hormone levels were１１．４mg/ ml and １０７ pg/ml, respectively. Chest computed tomography revealed an enhanced mass measuring approximately１．５cm located in the anterior mediastinum. ９９mTc-Methoxy-isobutylisonitrile scintigraphy demonstrated an anterior mediastinal mass. These findings suggested an ectopic parathyroid tumor located in the mediastinum. The patient underwent resection of the parathyroid tumor with video-assisted thoracic surgery（VATS）. The operation time was １１４ min. The postoperative day１（POD１）calcium level rapidly decreased to８．３mg/ml. The patient was discharged on POD５, and there have been no signs of recurrence １ year after the surgery. Parathyroidectomy by VATS for ectopic mediastinal parathyroid tumors is advantageous because it is less invasive and more cosmetic. VATS may be used as a standard approach for ectopic mediastinal parathyroid tumors. We report the surgical treatment of a case of ectopic mediastinal parathyroid adenoma using video-assisted thoracoscope

Usefulness of direct intratumoral administration of doxorubicin hydrochloride with an electro-osmosis–assisted pump

Patients receiving chemotherapy by intravenous (i.v.) or oral administration of anticancer drugs often experience side effects. In this study, an electro-osmotic flow (EO) pump was used for the direct administration of an anticancer drug with minimum side effects. Doxorubicin hydrochloride (DXR) was used as an anticancer drug, and its antitumor effect and toxicity were evaluated in comparison with i.v. administration. Balb/c female mice were subcutaneously transplanted with a breast cancer cell line (4T1/Luc) stably expressing luciferase, and 20 μL of DXR solution (1.0 mg/mL) was administered intratumorally (i.t.) at a slow rate (0.6 µL/min) using an EO pump or rapidly using a syringe. For comparison, 100 μL of DXR solution was injected through the tail vein at the same dose and a 5-times higher dose. A tumor growth inhibitory effect without significant weight loss was observed with direct i.t. administration of DXR using an EO pump. On the other hand, no suppressive tumor growth effect was observed with i.v. administration of DXR at the same dose. Although there was no significant difference in the suppression effect on tumor growth between i.t. administration with EO pump and syringe, the peripheral skin concentration of DXR were decreased after slow administration with EO pump compared with that after rapidly administration with a syringe. These results indicated that direct i.t. administration of DXR with lower dosing using an EO pump at slower administration rate may be useful for exhibiting antitumor effects and suppressing systemic side effects. In addition, the blood concentration and the peripheral skin concentration of DXR after administration at lower rate with EO pump were decreased compared with those after the rapidly administration with a syringe