Valosin-Containing Protein (VCP)/p97 is a key mediator between autopahgic cell death and apoptosis in adult hippocampal neural stem cells followin insulin withdrawal

Programmed cell death (PCD) plays essential roles in regulation of survival and function of neural stem cells (NSCs). Abnormal regulation of this process is associated with aging and neurodegenerative diseases. However, the mechanisms underlying the PCD of NSCs remain largely unknown. Therefore, understanding the mechanism of PCD in NSCs is crucial for exploring therapeutic strategy for the treatment of neurodegenerative diseases. We have previously reported that adult rat hippocampal neural stem (HCN) cells undergo autophagic cell death (ACD) following insulin withdrawal without apoptotic signs despite their normal apoptotic capabilities. It is unknown how interconnection between ACD and apoptosis is mediated in insulin-deprived HCN cells. Valosin-containing protein (VCP)/p97 is known to be essential for autophagosome maturation in mammalian cells. In this study, we report that VCP regulates the rate of autophagic flux in HCN cells following insulin withdrawal, suggesting the novel roles of VCP at other steps of autophagy as well as maturation. Particularly, VCP is expressed abundantly in HCN cells compared to hippocampal tissue and neurons. Pharmacological and genetic inhibition of VCP significantly decreased ACD and autophagy markers, while apoptotic cell death was induced in insulin-depleted HCN cells. Taken together, these data demonstrate that VCP may play an essential role in completion of ACD and mediation of crosstalk between ACD and apoptosis in HCN cells following insulin withdrawal. Elucidating the mechanism by which VCP regulates the crosstalk of ACD and apoptosis will contribute to understanding the molecular mechanism of PCD in NSCs. ⓒ 2015 DGIST1. INTRODUCTION 1 -- 2. Material and methods 3-- 2.1 Antibodies and reagents 3 -- 2.2 Cell culture 3 -- 2.3 Plasmids, siRNA and Transfection 3 -- 2.4 Cell death assay 4 -- 2.5 Western blotting 4 -- 2.6 Quantitative reverse transcription polymerase chain reaction (qRT-PCR) 5 -- 2.7 Annexin V staining and flow cytometry analysis 5 -- 2.8 Statistic analysis 5 -- 3. Results 6-- 3.1 VCP is degraded through autophagy in HCN cells following insulin withdrawal 6 -- 3.2 Inhibition of VCP switched autophagic cell death to apoptosis in insulin depleted HCN cells 6 -- 3.3 Inhibition of VCP significantly reduced autophagic flux in HCN cells following insulin withdrawal 7 -- 3.4 VCP regulates autophagy initiation signaling in insulin-deprived HCN cells 8 -- 4. Discussion 9 -- 5. Figure legends 11 -- 6. Figures 13 -- References 22 -- Abstract in Korean 24 -- Acknowledgement 25Programmed cell death (PCD)는 신경줄기세포의 생존과 기능조절에 필수적인 역할을 한다. 따라서 신경줄기세포의 비정상적인 PCD는 노화와 퇴행성 질환에 밀접한 관련이 있다고 보고되고 있다. 그러나, 신경줄기세포의 세포사멸기작에 대한 이해는 여전히 많은 부분이 밝혀져 있지 않아신경 줄기세포를 이용한 퇴행성 치료기술개발을 위한 큰 학문적 난제로 남아 있다. 그러므로, 신경줄기 세포에서 일어나는 PCD의 기작을 이해 하는 것은 노화와 퇴행성 질환의 치료 탐색을 위해 중요하다. 우리는 선행연구를 통해 성인 쥐의 해마성체신경줄기세포가 인슐린 결핍 조건에서 apoptotic 대신 순수한 autophagic cell death (ACD)를 겪는다는 것을 입증하였다. 성체해마신경줄기 세포는 정상적인 apoptosis 능력을 가지고 있기 때문에 인슐린이 결핍된 해마성체신경줄기세포에서왜 특이적으로 ACD가 유도되는지, 그리고 ACD와 apoptosis간의 상관관계는 어떻게 조절되는지는 아직 알려지지 않았다. Valosin-containing protein (VCP)는 포유류의 세포에서 autophagosome maturation에 필수적인 단백질이라고 알려져 있다. 본 연구에서는VCP가 인슐린이 결핍된 해마성체 신경줄기세포에서 autophagic flux의 속도를 조절한다는 것을 밝혔으며, 이는 autophagosome maturation뿐만 아니라 autophagy initiation을 포함한 ACD의 다른 과정에서 VCP의 새로운 역할들을 제시한다. 특이적으로, VCP는 hippocampal tissue와 hippocampal neuron 비해 해마성체신경줄기세포에 많이 발현했다. VCP의 약리학적 유전적인 억제는 인슐린이 결핍된 해마성체신경줄기세포에서 apoptosis가 유발 되는 동안 ACD와 autophagy markers를 감소시킴을 관찰하였다. 이러한 결과들은 VCP가 인슐린이 결핍된 해마성체신경줄기세포에서 ACD 유도와 ACD와 apoptosis 사이의 상호작용 조절에 중요한 역할을 할 것이라는 것을 입증하였다. VCP가 ACD와 apoptosis간의 상호작용을 조절하는 기작을 입증하는 것은 신경줄기세포에서 PCD의 분자적 기작을 이해하는데 도움을 줄 것이라 사료된다. ⓒ 2015 DGISTMasterdCollectio

Espaces d’expositions temporaires consacrés à l’art contemporain

Aux espaces conventionnels dédiés à l’exposition d’art contemporain se substituent des lieux alternatifs. Comme en témoigne plusieurs manifestations européennes, le monde de l’art s’ouvre au monde qui l’abrite. Tandis que la neutralité promue par le modèle du White Cube s’est imposé, apparaissent d’autres modes d’exposition le remettant en cause. Le lieu urbain non muséal occupe une place nouvelle et souvent centrale de l’œuvre : il devient un catalyseur de créativité.Conventional spaces dedicated to the presentation of contemporary art exhibitions are being replaced by alternative spaces. As is the case of several European initiatives here described, the art world opens up to the world in general. While the white cube model annihilated the presence of the mural space by neutralizing it, other exhibition modes have appeared who in turn challenge it. The non-museum urban place represents a new and often central spot for the artworks, while becoming a creativity catalyst

Histamine and Histamine H4 Receptor Promotes Osteoclastogenesis in Rheumatoid Arthritis.

Histamine H4 receptor (H4R) has immune-modulatory and chemotaxic effects in various immune cells. This study aimed to determine the osteoclastogenic role of H4R in rheumatoid arthritis (RA). The concentration of histamine in synovial fluid (SF) and sera in patients with RA was measured using ELISA. After RA SF and peripheral blood (PB) CD14+ monocytes were treated with histamine, IL-17, IL-21 and IL-22, and a H4R antagonist (JNJ7777120), the gene expression H4R and RANKL was determined by real-time PCR. Osteoclastogenesis was assessed by counting TRAP-positive multinucleated cells in PB CD14+ monocytes cultured with histamine, Th17 cytokines and JNJ7777120. SF and serum concentration of histamine was higher in RA, compared with osteoarthritis and healthy controls. The expression of H4R was increased in PB monocytes in RA patients. Histamine, IL-6, IL-17, IL-21 and IL-22 induced the expression of H4R in monocytes. Histamine, IL-17, and IL-22 stimulated RANKL expression in RA monocytes and JNJ7777120 reduced the RANKL expression. Histamine and Th17 cytokines induced the osteoclast differentiation from monocytes and JNJ7777120 decreased the osteoclastogenesis. H4R mediates RANKL expression and osteoclast differentiation induced by histamine and Th17 cytokines. The blockage of H4R could be a new therapeutic modality for prevention of bone destruction in RA

Pulsed Corona Discharge for Oxidation of Gaseous Elemental Mercury

Positive pulsed corona discharge has been applied for the oxidation of gaseous elemental mercury (Hg0) from a simulated flue gas. The oxidation of Hg0 to HgO and HgCl2 can significantly enhance the mercury removal from flue gas. At a gas condition of O2 (10%), H2O (3%), and N2 (balance), Hg0 oxidation efficiency of 84% was achieved at an input energy density of 45 J/l. The presence of NO, however, hinders Hg0 oxidation due to the preferential reaction of NO with O and O3. On the contrary, SO2 shows little effect on Hg0 oxidation due to its preferential reaction with OH. It has been also observed that the HCl in gas stream can be dissociated to Cl and Cl2 and can induce additional Hg0 oxidation to HgCl2

Mediastinal lymphoma in a young Turkish Angora cat

An 8-month old intact male Turkish Angora cat was referred to the Veterinary Medical Teaching Hospital (VMTH), Seoul National University, for an evaluation of anorexia and severe dyspnea. The thoracic radiographs revealed significant pleural effusion. A cytology evaluation of the pleural fluid strongly suggested a lymphoma containing variable sized lymphocytes with frequent mitotic figures and prominent nucleoli. The feline leukemia virus and feline immunodeficiency virus tests were negative. The cat was euthanized at his owner's request and a necropsy was performed. A mass was detected on the mediastinum and lung lobes. A histopathology evaluation confirmed the mass to be a lymphoma. Immunohistochemistry revealed the mass to be CD3 positive. In conclusion, the cat was diagnosed as a T-cell mediastinal lymphoma

Benserazide, the first allosteric inhibitor of Coxsackievirus B3 3C protease

AbstractCoxsackievirus B3 is the main cause of human viral myocarditis and cardiomyopathy. Virally encoded Coxsackievirus 3C protease (3Cpro) plays an essential role in viral proliferation. Here, benserazide was discovered as a novel inhibitor from a drug library screen targeting Coxsackievirus 3Cpro using a FRET-based enzyme assay. Benserazide, whose chemical structure has no electrophilic functional groups, was characterized as a non-competitive inhibitor by enzyme kinetic studies. A molecular docking study with benserazide and its analogs indicated that a novel putative allosteric binding site was involved. Specifically, a 2,3,4-trihydroxybenzyl moiety was determined to be a key pharmacophore for the enzyme’s inhibitory activity. We suggest that the putative allosteric binding site may be a novel target for future therapeutic strategies

A Study on ESD Protection Circuit Applying Silicon Controlled Rectifier-Based Stack Technology with High Holding Voltage

In this study, an improved Electrostatic Discharge (ESD) protection circuit with low trigger voltage and high holding voltage is proposed. ESD has become a serious problem in the semiconductor process because the semiconductor density has become very high these days. Therefore, much research has been done to prevent ESD. The proposed circuit is a stacked structure of the new unit structure combined by the Zener Triggering (SCR ZTSCR) and the High Holding Voltage SCR (HHVSCR). The simulation results show that the proposed circuit has low trigger voltage and high holding voltage. And the stack technology is applied to adjust the various operating voltage. As the results, the holding voltage is 7.7 V for 2-stack and 10.7 V for 3-stack