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The role of p21/CIP1/WAF1 (p21) in the negative regulation of the growth hormone/growth hormone receptor and epidermal growth factor/epidermal growth factor receptor pathways, in growth hormone transduction defect

  • Kostopoulou, Eirini;Gil, Andrea Paola Rojas;Spiliotis, Bessie E.
    • Annals of Pediatric Endocrinology and Metabolism
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    • v.23 no.4
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    • pp.204-209
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    • 2018
  • Purpose: Growth hormone transduction defect (GHTD) is characterized by severe short stature, impaired STAT3 (signal transducer and activator of transcription-3) phosphorylation and overexpression of the cytokine inducible SH2 containing protein (CIS) and p21/CIP1/WAF1. To investigate the role of p21/CIP1/WAF1 in the negative regulation of the growth hormone (GH)/GH receptor and Epidermal Growth Factor (EGF)/EGF Receptor pathways in GHTD. Methods: Fibroblast cultures were developed from gingival biopsies of 1 GHTD patient and 1 control. The protein expression and the cellular localization of p21/CIP1/WAF1 was studied by Western immunoblotting and immunofluorescence, respectively: at the basal state and after induction with $200-{\mu}g/L$ human GH (hGH) (GH200), either with or without siRNA CIS (siCIS); at the basal state and after inductions with $200-{\mu}g/L$ hGH (GH200), $1,000-{\mu}g/L$ hGH (GH1000) or 50-ng/mL EGF. Results: After GH200/siCIS, the protein expression and nuclear localization of p21 were reduced in the patient. After successful induction of GH signaling (control, GH200; patient, GH1000), the protein expression and nuclear localization of p21 were reduced. After induction with EGF, p21 translocated to the cytoplasm in the control, whereas in the GHTD patient it remained located in the nucleus. Conclusion: In the GHTD fibroblasts, when CIS is reduced, either after siCIS or after a higher dose of hGH (GH1000), p21's antiproliferative effect (nuclear localization) is also reduced and GH signaling is activated. There also appears to be a positive relationship between the 2 inhibitors of GH signaling, CIS and p21. Finally, in GHTD, p21 seems to participate in the regulation of both the GH and EGF/EGFR pathways, depending upon its cellular location.

Effects of Anticancer Drug Delivery based on Microbubble and Microbubble-Nanoparticle Complex using Low-Intensity Focused Ultrasound in Breast Cancer Animal Model (유방암 동물모델에서의 저강도 집속초음파를 이용한 마이크로버블 및 마이크로버블-나노물질 복합체 기반 항암제 전달 효율 검증)

  • Baek, Hee Gyu;Ha, Shin-Woo;Huh, Hyungkyu;Jung, Byeongjin;Han, Mun;Moon, Hyungwon;Kim, Sangkyun;Lee, Hak Jong;Park, Juyoung
    • Journal of Biomedical Engineering Research
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    • v.40 no.2
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    • pp.39-47
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    • 2019
  • Ultrasound sonication along with microbubble (MB) could enhance drug delivery to promote the absorption of anticancer drugs into cancers in a noninvasive and targeted manners. In this study, we verify the acute drug delivery enhancement (within an hour) of two representative focused ultrasound driven drug delivery enhancement methods (MB and Doxorubicin-coated Nanoparticle complex (MB-NP) based). Experiments were conducted using in vivo mouse model with MDA-MB-231 breast cancer cell line. Ultrasound generated by single-element 1 MHz focused ultrasound transducer was delivered in pulsed sonication consisted of 0.125 msec bursts at a pulse repetition frequency of 2 Hz for 20 seconds without a significant increase in local temperature (less than $0.1^{\circ}C$) or hemorrhage. Doxorubicin concentrations in tumors were improved by 1.97 times in the case of MB-NP, and 1.98 times by using Doxorubicin and MB separately. These results indicate anticancer drug delivery based on MB and MB-NP can significantly improve the effect of anticancer drugs delivered to tumors in a short time period by using low-intensity focused ultrasound.

A Study on Vibration & Noise Reduction of Fast Back Feeding Device for Manufacturing Process (제조공정용 Fast Back 이송장치 진동·소음 저감에 관한 연구)

  • Han, Doo-Hee;Lee, Seung-Hun;Son, Yung-Deug
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.8
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    • pp.642-648
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    • 2019
  • This paper presents a fast back-type transfer device for snack food processing that uses the inertia of transferred material. A conventional conveying system is a drive system that uses a belt conveyor and mechanical crank, which generate noise and vibration and cause environmental pollution. Vibration and noise are reduced in the proposed fast back feeding device by using a counterweight. The crank drive unit was replaced with a linear servomotor, and an equilibrium device was designed to balance the force due to acceleration. This makes it is possible to adjust the forward and backward speed and acceleration through PLC control. A vibration damper device offsets the vibration force of the periodic shock form. The main cause of the vibration was identified through vibration analysis, and reduction measures were established. We verified the effectiveness of the vibration by making a prototype and performing about 10 vibration tests. Because no mechanical transducer is needed, energy loss, noise, and vibration do not occur, and the operating speed is not limited.

Waveform characteristics of ultrasonic wave generated from CNT/PDMS composite (CNT/PDMS 복합체로부터 방사된 초음파의 파형 특성)

  • Kim, Gisuk;Kim, Moojoon;Ha, Kanglyeol;Lee, Jooho;Paeng, Dong-Guk;Choi, Min Joo
    • The Journal of the Acoustical Society of Korea
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    • v.38 no.4
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    • pp.459-466
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    • 2019
  • When a laser pulse is irradiated on a CNT (Carbon Nanotube) and PDMS (Poly dimethylsiloxane) composite coated on a transparent PMMA (Poly methyl methacrylate) substrate, a strong ultrasonic wave is generated due to the thermoelastic effect. In this paper, the thermoacoustic theory related to the wave generation by the CNT/PDMS composite was established. The waveforms of ultrasonic waves when a laser pulse having a Gaussian waveform is irradiated on the composite with a thickness of $20{\mu}m$ were numerically simulated. From the results, it was confirmed that ultrasonic shock waves can be generated from the CNT/PDMS composite and the waveforms are changed little even if the physical properties of the composite are changed by ${\pm}20%$. It was found that the peak positive and negative pressures increase as the thermal expansion coefficient increases, or as density, heat capacity and sound speed decreased. However, those changes were not so sensitive with thermal conductivity. In addition, the physical properties of the CNT/PDMS composite fabricated in this study were estimated from the comparison of the measurement and simulation results.

Effect of serotonin on the cell viability of the bovine mammary alveolar cell-T (MAC-T) cell line

  • Xusheng, Dong;Chen, Liu;Jialin, Miao;Xueyan, Lin;Yun, Wang;Zhonghua, Wang;Qiuling, Hou
    • Journal of Animal Science and Technology
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    • v.64 no.5
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    • pp.922-936
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    • 2022
  • 5-Hydroxytryptamine (5-HT), a monoamine, as a local regulator in the mammary gland is a chemical signal produced by the mammary epithelium cell. In cows, studies have shown that 5-HT is associated with epithelial cell apoptosis during the degenerative phase of the mammary gland. However, studies in other tissues have shown that 5-HT can effectively promote cell viability. Whether 5-HT could have an effect on mammary cell viability in dairy cows is still unknown. The purpose of this study was to determine: (1) effect of 5-HT on the viability of bovine mammary epithelial cells and its related signaling pathways, (2) interaction between prolactin (PRL) and 5-HT on the cell viability. The bovine mammary alveolar cell-T (MAC-T) were cultured with different concentrations of 5-HT for 12, 24, 48 or 72 hours, and then were assayed using cell counting kit-8, polymerase chain reaction (PCR) and immunobloting. The results suggested that 20 μM 5-HT treatment for 12 or 24 h promote cell viability, which was mainly induced by the activation of 5-HT receptor (5-HTR) 1B and 4, because the increase caused by 5-HT vanished when 5-HTR 1B and 4 was blocked by SB224289 and SB204070. And protein expression of mammalian target of rapamycin (mTOR), eukaryotic translation elongation factor 2 (eEF2), janus kinase 2 (JAK2) and signal transducer and activator of transcription 5 (STAT5) were decreased after blocking 5-HT 1B and 4 receptors. When MAC-T cells were treated with 5-HT and PRL simultaneously for 24 h, both the cell viability and the level of mTOR protein were significantly higher than that cultured with 5-HT or PRL alone. In conclusion, our study suggested that 5-HT promotes the viability of MAC-T cells by 5-HTR 1B and/or 4. Furthermore, there is a reciprocal relationship between PRL and 5-HT.

Studies on the regulation of Hematopoietic enhancement of Brassica campestris var narinosa., Canavalia gladiata DC semen and their combinational prescription via Jak2/STAT5/GATA1 Pathway in Sca-1+ hematopoietic stem cells (Sca-1+골수조혈세포에서 JAK2/STAT5/GATA-1 신호전달 경로를 통한 다채, 도두 그리고 두 조합물에 의한 조혈증진 조절에 관한 연구)

  • Kim, Kunhoae;Kim, Seung-Hyung;Cho, In-Sik;Kim, Han-Young;Kim, Dong-Seon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.7-16
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    • 2013
  • Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-$1^+$ hematopoietic stem cells (Sca-$1^+HSCs$) as a testing system. Methods : Sca-$1^+HSCs$ isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of $CD117^+/Sca-1^+$ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When Sca-$1^+HSCs$ were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-$1^+HSCs$. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-$1^+HSCs$. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.

Measurement of sonoluminescence intensity for evaluation of the amount of radical generated by ultrasonic cavitation (초음파 캐비테이션에 의해 생성되는 라디칼의 발생량 평가를 위한 소노루미네센스 발광강도의 측정)

  • Jungsoon Kim;Moojoon Kim
    • The Journal of the Acoustical Society of Korea
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    • v.42 no.1
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    • pp.50-56
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    • 2023
  • The hydroxyl radical (·OH) and superoxide anion radical (·O2- ) generated by the shock wave generated during ultrasonic cavitation collapse in TiO2 suspension are highly useful because they can sterilize and disinfect. For practical use as a sterilization method without any chemicals, in this study, we proposed a method for evaluating the generation of radicals generated by high-intensity ultrasound emitted to titanium dioxide suspension. In the proposed method, the sonoluminescence phenomenon, which emits light by ultrasonic cavitation decay energy, was utilized, and the degree of radical generation was evaluated through the amount of light energy by sonoluminescence. As a result, even at a low concentration of titanium dioxide of 0.02 wt%, light energy 5 times higher than in the absence of titanium dioxide was received. After that, as the concentration increased by 0.1 wt%, the luminous intensity of sonoluminescence increased linearly by about 14.8 × 10-12 lm. Therefore, it was confirmed that the radicals generated by radiating high-intensity ultrasound to the titanium dioxide suspension increased linearly as the concentration of titanium dioxide increased within a given concentration range.

Experiment and Analysis of Backscattering Signals According to Presence or Absence of Chloroform (클로로폼 침적 유무에 따른 후방산란신호 측정 실험 및 분석)

  • Him Chan Seo;Jee Woong Choi;Yongmyung Kim;Moonjin Lee
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.28 no.spc
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    • pp.18-22
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    • 2022
  • Because it is difficult to apply direct and optical detection techniques to sunken hazardous and noxious substances (HNS), effective acoustic detection techniques are required to detect sunken HNS in water. In this study, the possibility of acoustic detection of sunken HNS was investigated through backscattering signal measurement experiments using chloroform, a sunken HNS. After establishing a pool in an acrylic tank, backscattering signals were measured according to the presences or absence of chloroform by varying the grazing angle from 90° to 50° in 0.5° intervals using a pan&tilt system. A directional transducer transmitted and received sinusoidal signals with a frequency of 200 kHz and a pulse length of 25 ㎲ in a monostatic state. When chloroform was deposited, the received level of the backscattering signal at the interface between water and chloroform became low at a grazing angle of approximately 80° or smaller. Based on the backscattering signal results obtained at the interface between water and chloroform, the possibility of acoustic detection of sunken HNS was demonstrated.

Evaluation of Immune Enhancing Activity of Luthione, a Reduced Glutathione, in RAW 264.7 Macrophages (RAW 264.7 대식세포에서 환원형 glutathione인 luthione의 면역 증강 활성 평가)

  • Seon Yeong Ji;Da Hye Kwon;Hye Jin Hwang;Yung Hyun Choi
    • Journal of Life Science
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    • v.33 no.5
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    • pp.397-405
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    • 2023
  • Although glutathione (GSH) has been shown to play an important role in the prevention of oxidative damage as an antioxidant, studies on immune regulation by it have not been properly conducted. In this study, we investigated whether luthione®, a reduced GSH, has an immune enhancing effect in murine macrophage RAW 264.7 cells. The results of flow cytometry and immunofluorescence experiments indicated that luthione increased phagocytic activity, a representative function of macrophages, compared to the control cells. According to the results of the cytokine array, the expression of interleukin (IL)-5, IL-1β, and IL-27 was significantly increased in the luthione-treated cells. Luthione also enhanced the production of tumor necrosis factor-α and IL-1β through increased expression of their proteins, and increased release of the immune mediators such as nitric oxide (NO) and prostaglandin E2 was associated with increased expression of inducible NO synthase and cyclooxygenase-2. In addition, the expression of cluster of differentiation 86, an M1 macrophage marker, was dramatically enhanced in RAW 264.7 cells treated with luthione. Furthermore, as a result of heat map analysis, we found that cytokine signaling 1/3-mediated signal transducer and activator of transcription/Janus tyrosine kinase signaling pathway was involved in the immunomodulatory effect by luthione. In conclusion, our data suggested that luthione could act as a molecular regulator in M1 macrophage polarization and enhance immune capacity by promoting macrophage phagocytic function.

Guide for Processing of Textured Piezoelectric Ceramics Through the Template Grain Growth Method

  • Temesgen Tadeyos Zate;Jeong-Woo Sun;Nu-Ri Ko;Hye-Lim Yu;Woo-Jin Choi;Jae-Ho Jeon;Wook Jo
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.36 no.4
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    • pp.341-350
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    • 2023
  • The templated grain growth (TGG) method has gained significant attention for its ability to produce highly textured piezoelectric ceramics with significantly enhanced performance, making it a promising method for transducer and actuator applications. However, the texturing process using the TGG method requires the optimization of multiple steps, which can be challenging for beginners in this field. Therefore, in this tutorial, we provide an overview of the TGG method mainly based on our previous published works, including its various processing steps such as synthesizing anisotropic-shaped templates with size and size distribution control using the molten salt synthesis technique, tape casting, and identifying key factors for proper alignment of the templates in the target matrix system. Our goal is to provide a resource that can serve as a basic reference for researchers and engineers looking to improve their understanding and utilization of the TGG method for producing textured piezoelectric ceramics.