Selective anti-cancer activity of Hirsutine against HER2 positive breast cancer cells by inducing DNA damage

Hirsutine is one of the major alkaloids isolated from plants of the Uncaria genus and is known for its cardioprotective, anti‑hypertensive and anti-arrhythmic activities. We recently reported that hirsutine is an anti-metastatic phytochemical by targeting NF-κB activation in a murine breast cancer model. In the present study, we further examined the clinical utility of hirsutine against human breast cancer. Among six distinct human breast cancer cell lines, hirsutine showed strong cytotoxicity against HER2-positive/ p53-mutated MDA-MB‑453 and BT474 cell lines. Conversely, HER2-negative/p53 wild‑type MCF-7 and ZR-75-1 cell lines showed resistance against hirsutine-induced cytotoxicity. Hirsutine induced apoptotic cell death in the MDA-MB-453 cells, but not in the MCF-7 cells, through activation of caspases. Furthermore, hirsutine induced the DNA damage response in the MDA-MB-453 cells, but not in the MCF-7 cells, as highlighted by the upregulation of γH2AX expression. Along with the induction of the DNA damage response, the suppression of HER2, NF-κB and Akt pathways and the activation of the p38 MAPK pathway in the MDA-MB-453 cells were observed. Considering that there was no difference between MDA-MB-453 and MCF-7 cells in regards to irinotecan‑induced DNA damage response, our present results indicate the selective anticancer activity of hirsutine in HER2-positive breast cancer by inducing a DNA damage response

Estimation of springback of stainless steel sheet part taking influence of anisotropic property of plastic-deformation-dependent young's modulus into account

A kinematic hardening model proposed by Yoshida and Uemori (Y-U model) was applied to the prediction of springback of stainless steel sheet part. From the experiments for the determination of the material constants, an anisotropic property of change in Young’s modulus was observed; namely, the anisotropy was different at 0°, 45° and 90° from the rolling direction. The Y-U model for the stainless steel sheet was used to a calculation of a forming process of a part to examine the accuracy of the prediction of the springback by compar-ing the calculated result with the actual part formed. In order to consider the anisotropic property of change in Young’s modulus, the calculated result to the actual part formed. In order to consider the anisotropic property of the change in Young’s modulus, the calculations were performed using the different material constants at 0°, 45° and 90° from the rolling direction. With the material constants at 90° from the rolling direction, which was the direction of springback of the part, the prediction accuracy can be improved. Therefore, the consideration of the anisotropic property of the change in Young’s modulus was found to be effective for more accurate prediction of the springback of the stainless steel part

フォンヴィレブランド因子に存在する血型A抗原は、ADAMTS13による切断に対してB・H抗原よりも抵抗性を示す

BACKGROUND: ADAMTS13 specifically cleaves the peptide bond between Y1605 and M1606 within the VWF-A2 domain. OBJECTIVE: The VWF contains ABO(H) blood group antigens, which may influence the susceptibility of VWF to ADAMTS13. METHODS: Using a unique monoclonal antibody recognizing the Y1605 residue, we have developed a sandwich ELISA to analyze the generation of a VWF-DP by ADAMTS13 quantitatively. RESULTS: Production of VWF-DP after exposure to four different degrees of high shear stress was validated in comparison to the reduction in high-molecular-weight multimers using VWF multimer analysis. In analysis of plasma from 259 healthy individuals, plasma levels of VWF antigen (VWF:Ag) were significantly lower in blood group O than in the other groups and were significantly correlated with plasma VWF-DP levels. The ratio between VWF-DP and VWF:Ag was significantly higher in blood group O than in blood groups A and AB. The ratio in blood group B was also significantly higher than those in A and AB, but did not differ from blood group O. Finally, to examine whether ABO(H) blood group antigens contributed to VWF cleavage, 82 plasma samples were exposed to high shear stress using a cone-plate shear stress inducer. The difference in the VWF-DP/VWF:Ag ratio before and after high shear stress in blood group O was significantly greater than those in groups A and AB. CONCLUSION: These results indicate that blood group antigen A on VWF was more protective against ADAMTS13 cleavage than antigens B and H.博士（医学）・乙第1440号・令和元年9月27日© 2019 International Society on Thrombosis and HaemostasisThis is the pre-peer reviewed version of the following article: [https://onlinelibrary.wiley.com/doi/abs/10.1111/jth.14444], which has been published in final form at [https://doi.org/10.1111/jth.14444]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions

クリオ上清中の高分子VWFマルチマー非結合型ADAMTS13の存在:血栓性血小板減少性紫斑病の治療に、より効果的な血漿分画製剤の選択

BACKGROUND: Thrombotic thrombocytopenic purpura (TTP) is characterized by deficient ADAMTS13 activity. Treatment involves plasma exchange (PE). Both fresh-frozen plasma (FFP) and cryosupernatant (CSP) are used, but it remains to be determined which is more effective. STUDY DESIGN AND METHODS: To analyze the interaction between von Willebrand factor (VWF) and ADAMTS13, we used large-pore isoelectric focusing (IEF) analysis followed by detection with anti-ADAMTS13 monoclonal antibody. FFP, CSP, cryoprecipitate (CP), and purified ADAMTS13 were analyzed for their effects on high shear stress-induced platelet aggregation (H-SIPA). RESULTS: IEF analysis of normal plasma revealed three groups of ADAMTS13 bands with pI of 4.9 to 5.6, 5.8 to 6.7, and 7.0 or 7.5. Two band groups (pI 4.9-5.6 and 5.8-6.7) were found in plasma of a patient with Type 3 von Willebrand disease, in which VWF is absent, whereas no bands were found in plasma of a patient with congenital ADAMTS13 deficiency. Mixing these plasmas generated the bands at pI 7.0 or 7.5, representing the VWF-ADAMTS13 complex; these bands were absent in CSP. FFP and purified ADAMTS13 down regulated H-SIPA in a dose-dependent manner. However, CP did not inhibit H-SIPA in the initial phase, and the degree of inhibition at the endpoint was almost indistinguishable from those of the other two plasma products. CONCLUSION: Both plasma products (FFP and CSP) are effective for PE in TTP patients. However, CSP may be more favorable, because it has lower levels of VWF and almost normal ADAMTS13 activity, but lower levels of ADAMTS13 in complex with larger VWF multimers.博士（医学）・乙第1322号・平成25年11月27日© 2013 American Association of Blood BanksCopyright © 1999-2018 John Wiley & Sons, Inc. All rights reservedThis is the pre-peer reviewed version of the following article: [https://onlinelibrary.wiley.com/doi/full/10.1111/trf.12182], which has been published in final form at [http://dx.doi.org/10.1111/trf.12182]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions

Immunopharmacological properties of Oren-gedoku-to (a Kampo medicine, Huang-Lian-Jie-Du-Tang) on contact hypersensitivity reaction in mice

We investigated the effects of Oren-gedoku-to (Huang-Lian-Jie-Du-Tang), a Kampo medicine, on DNFB-induced contact hypersensitivity (CHS) response in mice in order to further clarify the immunopharmacological properties of this formulation. 1) Administration of Oren-gedoku-to decreased the magnitude of ear swelling in the CHS response and shortened the affected period. The inhibitory effect on ear swelling was observed even when Oren-gedoku-to was given orally with different timing schedules. 2) The expressions of mRNAs for CD8, IFN-7 and TNF-α in the ear of Oren-gedoku-to-treated mice were markedly decreased 24 h after the challenge. 3) The number of skin-draining regional lymph node cells (LNCs), CD4^+ T cells and CD8^+ T cells was decreased without affecting the ratio of CD8^+/CD4^+ T cells. 4) Oren-gedoku-to resulted in a marked impairment of the hapten-specific development of LNCs. These results suggest that the suppressive effect of Oren-gedoku-to on ear swelling was partly caused by the suppression of lymphocyte proliferation. 接触過敏反応(CHS)に対する黄連解毒湯の抑制効果について検討した。1g/kgの黄連解毒湯を感作日より連続投与することで,DNFB塗布による耳介の腫脹は軽減し,その持続時間も短縮した。また,黄連解毒湯の投与期間を変更(感作後0-2日間あるいは4-6日間の投与)しても抑制効果が認められた。耳介局所では,黄連解毒湯の連続投与により,CD8,IFN-γおよびTNF-αのmRNA発現は減弱した。所属リンパ節では,全リンパ節細胞,CD8^+T細胞,CD4^+T細胞の数が減少したが,CD8/CD4比に変化はみられなかった。さらに,リンパ節細胞のハプテン特異的な増殖能は抑制された。以上の結果より,黄連解毒湯のCHSの抑制効果にハプテン特異的リンパ球の増殖抑制が関与していると考えられた

重度大動脈弁狭窄症患者の大動脈弁置換術後における血小板機能および高分子量 von Willebrand 因子多量体の急速な回復

AIM: Patients with severe aortic stenosis (AS) may have bleeding episodes due to the loss of high-molecular-weight (HMW) von Willebrand factor multimers (VWFMs). The absence of HMW-VWFMs and bleeding tendency are usually corrected after aortic valve replacement (AVR). To investigate the process of VWFM recovery and symptoms in patients with severe AS, we analyzed changes in VWF antigen (VWF:Ag), ADAMTS13 activity (ADAMTS13:AC), and platelet thrombus formation under high shear stress conditions. METHODS: Nine patients with severe AS undergoing AVR were analyzed. RESULTS: Evident deficiency of HMW-VWFMs was observed in six patients before surgery, which was rapidly restored within 8 days after AVR. Median levels of VWF:Ag before surgery, on postoperative days (PODs) 1, 8, 15, and 22, and one year after AVR were 78.1%, 130%, 224%, 155%, 134%, and 142%, respectively. In contrast, ADAMTS13:AC was 50.5%, 35.5%, 25.5%, 25.1%, 30.3%, and 84.6%, respectively. Preoperative thrombus formation but not surface coverage was significantly lower than that on POD 22, which was considered as normal level in each patient. Compared with preoperative levels, thrombus volume was significantly lower on POD 1, but rapidly increased by POD 8. CONCLUSION: Bleeding tendency and loss of HMW-VWFMs observed in patients with severe AS before surgery was rapidly corrected after AVR. Instead, patients were in a VWF-predominant state between POD 8 and 22.博士（医学）・乙第1395号・平成29年3月15日Copyright © 2016 Japan Atherosclerosis Society本論文の著作権は日本動脈硬化学会が保持しています。This article is distributed under the terms of the latest version of CC BY-NC-SA defined by the Creative Commons Attribution License

Mammary tissue microenvironment determines T cell-dependent breast cancer-associated inflammation

Although the importance of the host tissue microenvironment in cancer progression and metastasis has been established, the spatiotemporal process establishing a cancer metastasis-prone tissue microenvironment remains unknown. In this study, we aim to understand the immunological character of a metastasis-prone microenvironment in a murine 4T1 breast tumor model, by using the activation of nuclear factor-jb (NF-jB) in cancer cells as a sensor of inflammatory status and by monitoring its activity by bioluminescence imaging. By using a 4T1 breast cancer cell line stably expressing an NF-jB ⁄ Luc2 reporter gene (4T1 NF-jB cells), we observed significantly increased bioluminescence approximately 7 days after metastasis-prone orthotopic mammary fat-pad inoculation but not ectopic s.c. inoculation of 4T1 NF-jB cells. Such in vivo NF-jB activation within the fat-pad 4T1 tumor was diminished in immune-deficient SCID or nude mice, or T celldepleted mice, suggesting the requirement of host T cell-mediated immune responses. Given the fat-pad 4T1 tumor expressed higher inflammatory mediators in a T cell-dependent mechanism compared to the s.c. tumor, our results imply the importance of the surrounding tissue microenvironment for inflaming tumors by collaborating with T cells to instigate metastatic spread of 4T1 breast cancer cells

Effect of keishibukuryogan on genetic and dietary obesity models

Obesity has been recognized as one of the most important risk factors for a variety of chronic diseases, such as diabetes, hypertension/cardiovascular diseases, steatosis/hepatitis, and cancer. Keishibukuryogan (KBG, Gui Zhi Fu Ling Wan in Chinese) is a traditional Chinese/Japanese (Kampo) medicine that has been known to improve blood circulation and is also known for its anti-inflammatory or scavenging effect. In this study, we evaluated the effect of KBG in two distinct rodent models of obesity driven by either a genetic (SHR/NDmcr-cp rat model) or dietary (high-fat diet-induced mouse obesity model) mechanism. Although there was no significant effect on the body composition in either the SHR rat or the DIO mouse models, KBG treatment significantly decreased the serum level of leptin and liver TG level in the DIO mouse, but not in the SHR rat model. Furthermore, a lower fat deposition in liver and a smaller size of adipocytes in white adipose tissue were observed in the DIO mice treated with KBG. Importantly, we further found downregulation of genes involved in lipid metabolism in the KBG-treated liver, along with decreased liver TG and cholesterol level. Our present data experimentally support in fact that KBG can be an attractive Kampo medicine to improve obese status through a regulation of systemic leptin level and/or lipid metabolism

NKG2D function protects the host from tumor initiation

The activation NKG2D receptor has been shown to play an important role in the control of experimental tumor growth and metastases expressing ligands for NKG2D; however, a function for this recognition pathway in host protection from de novo tumorigenesis has never been demonstrated. We show that neutralization of NKG2D enhances the sensitivity of wild-type (WT) C57BL/6 and BALB/c mice to methylcholanthrene (MCA)-induced fibrosarcoma. The importance of the NKG2D pathway was additionally illustrated in mice deficient for either IFN-γ or tumor necrosis factor–related apoptosis-inducing ligand, whereas mice depleted of natural killer cells, T cells, or deficient for perforin did not display any detectable NKG2D phenotype. Furthermore, IL-12 therapy preventing MCA-induced sarcoma formation was also largely dependent on the NKG2D pathway. Although NKG2D ligand expression was variable or absent on sarcomas emerging in WT mice, sarcomas derived from perforin-deficient mice were Rae-1+ and immunogenic when transferred into WT syngeneic mice. These findings suggest an important early role for the NKG2D in controlling and shaping tumor formation

An Empirical Assessment of the Business Value Derived from Implementing Mobile Technology: A Case Study of Two Organizations

Mobile technologies are argued to offer unprecedented opportunities for organizations and individuals. In order for organizations to be persuaded that investment in mobile technologies is not only worthwhile, but also important to the achievement of corporate goals and objectives, then it is important to evaluate the potential of mobile technology so that the derivation of business value and the related risks involved in implementing mobile devices and services in an organization can be understood. This paper aims at understanding the organizational value that could be derived from investments in mobile technology. We present two in-depth case studies of mobile technology implementation in health care organizations. These studies show that deriving business value from the adoption and implementation of mobile devices does not seem at all certain, but is contingent upon clear business objectives and a willingness to make business changes to embrace the transformation to core business processes which are driven by the mobile technologies