Synegistic Anti-proliferation Effects of All -trans Retinoic Acid with Paclitaxel on Autosomal Polycystic Kidney Disease Cell Line

NGUYEN THANH THANH QUE Graduate School Gachon University 2020 국내석사

ABSTRACT Synergistic anti-proliferation effects of all-trans retinoic acid with paclitaxel on autosomal polycystic kidney disease cell line. Que Thanh Thanh Nguyen Supervisor: Prof. Jae Young Kim Department of Life Science Graduate School of Gachon University Autosomal dominant polycystic kidney disease (ADPKD) has been well-known as a monogenic inherited disorder and characterized by cyst development in the kidney. In recent years, the mechanism in autosomal dominant polycystic kidney disease (ADPKD) associated to genetic factors and non-genetic factors have been widely studied to get deep knowledge about ADPKD. In renal tubules, renal epithelial cells over-growth due to lower intracellular calcium level and the mitogenic effect of cAMP (cyclic adenosine monophosphate), is one of the hallmark features that originated cystogenesis. In this study, for the first time, we investigated the synergistic effects of combinatory treatment between all-trans retinoic acid (ATRA), an active form of vitamin A, and paclitaxel, a high effective anticancer drug, on the autosomal dominant polycystic kidney cell line. Cells were treated with a combination of ATRA with paclitaxel for 48 h and then analyzed by cell viability assay. In combinatory treatment groups, our data indicated that the percentage of cell viability was significantly decreased in the comparison with DMSO (control) and ATRA/paclitaxel (alone) treatment. Cell cycle arrest showed that the combinatory treatment of ATRA with paclitaxel induce ADPKD cell line arrest at the G2/M phase. Furthermore, apoptotic cells were detected in our study by Annexin V/PI staining. The up-regulation of p53 protein expression and caspase activation could be involved in apoptosis signal and cell cycle arrest. We revealed that the intra-calcium level was sharply increased by combined treatment of ATRA and paclitaxel after 72 h of treatment and this enhance was blocked by RARα/RXRα antagonist or calcium channel blockers. Besides, ADPKD cell proliferation was controlled through the ERK signaling pathway by in phosphorylated ERK. Our result suggested the function of combinatorial treatment of ATRA with paclitaxel which expresses synergistic anti-proliferation effect on ADPKD cell proliferation by inducing cell cycle arresting, apoptosis and suppressing the ERK signaling pathway. Keywords: Autosomal polycystic kidney disease (ADPKD), ATRA, paclitaxel, cell cycle arrest, apoptosis, intracellular calcium, phosphorylated ERK1/2.

Preventive Effects of Fermented Cordyceps militaris on Obesity and Hepatosteatosis

TRANSONGNGUYENKHOI Gachon University, Gachon University 2018 국내박사

Various studies have shown that nonalcoholic fatty liver diseases (NAFLD), the aggregation of lipid droplets in the liver (hepatosteatosis) followed by inflammatory complication, appears to be chronic consequences of overnutrition. In the present research, we examined the anti-hepatosteatosis effects of fermented Cordyceps militaris (CM) extracts in AML-12 hepatocytes as well as in mice with short-term high-fat diet (HFD) via activation of fatty acid oxidation (FAO). The chemical components, adenosine and cordycepin, of CM extract (CME), CM grown on germinated soybean (GSCE) and fermented GSC by Pediococcus pentosaceus ON188 (ON188E) were assessed. In hepatocytes, the expression of CPT1α and PPARα, FAO genes were upregulated only by ON188E treatment. In contrast, SCD1, a lipogenic gene was downregulated by ON188E within no cytotoxic range. Regardless of oleic acid treatment to mimic fatty liver condition, mRNA and proteins of FAO genes were upregulated by ON188E in a dose-dependent manner. Formation of intracellular lipid droplets by addition of oleic acid was reduced by ON188E to the levels in WY14643-treated cells. When cells were treated with ON188E, SPHK2 and its phosphorylated product sphingosine 1-phosphate (S1P) elevation activated FAO in hepatocytes. In consistent with this, in animal results, ON188E extract did not alter the food intake while body weight and inflammatory factors in plasma of HFD mice were controlled comparable to those of NCD mice. Moreover, ON188E treatment significantly upregulated hepatic SPHK2 level in mice fed HFD. The elevated SPHK2 activated hepatic FAO genes such as CPT1α and PPARα but downregulated DGAT2, a triglyceride biosynthetic gene. We found that ON188E treatment reduced lipid droplets in liver and the hypertrophy of adipocytes in epididymal white adipose tissue (WAT) in a dose-dependent manner. Activation of SIRT1and PPARγ in WAT suggests that ON188E could modulate browning process of adipocytes which induces body thermogenesis. Collectively, the fermented CM extracts not only reduce risk of tissue damage but activate hepatic FAO via elevation of S1P synthesis in hepatocytes and induce WAT browning as well. This suggests ON188E can be applied as a potential functional food preventing obesity and hepatosteatosis.

FUNCTIONALIZATION OF CUPROUS OXIDE NANOPARTICLES FOR PHOTOCATALYTIC ACTIVITY

Mohit Kumar Gachon University 2017 국내석사

The amino acids, namely L-glutamic acid and glucosamine were used as stabilising agents by a wet chemical method for the synthesis of Cu2O nanoparticles, where -NH2 groups attached through coordinated bonds with Cu2+ ions homogeneously. This procedure is a simple route to synthesize nanoparticles without high pressure, high temperature and a long period of time. Here, L-ascorbic acids can control the reduction of Cu2+ ions to Cu1+ ions in an aqueous solution at room temperature. Synthesised nanomaterial were confirmed by powder X-ray diffraction (XRD), Fourier transform infrared (FTIR), scanning electron microscope (SEM) and Transmission electron microscopy (TEM). In addition, these functionalized cuprous oxide nanoparticles exhibited excellent absorption properties in the visible light irradiation. The reasonably low priced cuprous oxide (Cu2O) nanoparticles are known as a p-type semiconductor and has a potential photocatalytic degradation activity of MB in aqueous solution. The contribution of invariant electron acceptor H2O2 increases the reaction rate of photocatalytic activity due to the formation of more OH* free radicals. The dye degradation proceeds with high rate constant value and the functionalised materials show good stability and reuse capability. In the second work, discuss about metal ions sensing by fluorometric technique using Zinc Oxide Nanowires. ZnO NWs were synthesized at low temperature by simple precipitation method. This modest to prepare ZnO NWs giving a convenient and reliable sensing of Cu (II) and Cr (VI) ions in useful application. A unique methodology relies from the coordination with ZnO nanowires. The concentration of copper and chromium were determined from decreasing in photoluminescence intensity at room temperature.

Protective Effects of Morning Glory Seeds against Nephrotoxicity in vitro and in vivo

Yanli Shao Gachon University 2017 국내석사

The renal-protective effects of morning glory seed extract (MGs-Hx) were evaluated in the HEK-293 cell line and ICR mice in which nephrotoxicity or acute kidney injury were induced using cisplatin. HEK-293 cells were co-treated in vitro with MGs-Hx (2 h) and cisplatin (24 h) and cell viability, nitric oxide, apoptosis, inflammation, and apoptosis-related proteins were evaluated. My results showed that MGs-Hx treatment increased HEK-293 cell viability compared with cisplatin (5 μM) alone. MGs-Hx also significantly reduced the expression of the inflammatory enzyme iNOS and the continuous production of NO. In addition, MGs-Hx (90 μg/ml or 100 μg/ml) exhibited a tendency to decrease apoptosis-related caspase-3, -8, and -9 protein levels as well as promote IB-α degradation and NF-B translocation to the nucleus in cisplatin nephrotoxic HEK-293 cells. MGs-Hx showed remarkable inhibition of MAPK activation (p38, ERK, and JNK) in cisplatin-induced HEK-293 cells. In vivo, ICR male mice were orally administered MGs-Hx, 2 h before intraperitoneal injection of cisplatin. The mice were sacrificed after 7 days, and their blood and kidney samples were collected for analysis. MGs-Hx (100 mg/kg) showed highly protective effects with low side effects, significant decrease in serum creatinine, blood urea nitrogen, and serum uric acid levels, and reduced expression of iNOS and COX-2 mRNA. These findings suggest that MGs-Hx has potential renoprotective effects at low concentrations.

Comparison Analysis of Accessibility Infrastructure for Elderly and Disabled Between South Korea Subway and Jakarta Commuter Railway Station

Aryanti, Riris Aryanti Gachon University Graduate School 2016 국내석사

Accessibility is about giving equal access to everyone and without being able to access facilities and services, a person with disabilities will never be fully included (United Nation, 2007). The friendly environment is the one of human prospect life, accessibility becomes the real fact to create sustainability environment fulfill people mobility needs. Proper access is important to support their availability access, happily and independently life. Most of the public facilities in South Korea and Western countries have provided proper access for elderly and disabled. Unlike Indonesia, accessibility standard has not been applied appropriately. The Aim of this research is highlighting the assesment result of railway station infrastructure by using Rank Weighted Method and SWOT analysis to know the affect factor for create the strategic action plan regarding accessibility railway infrastructure for elderly and disabled. Create the friendly and inclusive environment by give the best infrastructure design standard of Jakarta Commuter Railway public services accessible to their beneficiaries disregarding their physical, mental or language limitations and urging the needs of improving safety awareness and empowering the disabled society eith more access to using rail commuter railway in Jakarta.

Effects of glucagon-like peptide-1 receptor agonist on disuse muscle atrophy

Nguyen, Tram Thi Ngoc Gachon University 2019 국내석사

Berenil: A Selective Anti-bacterial Agent for E.coli O157: H7

Wu, Siying Gachon University 2015 국내석사

This study investigated the bacteriostatic and bactericidal effects of Berenil against 5 strains of pathogenic bacteria and 2 strains of non-pathogenic bacteria. The results showed that under optimized conditions (temperature and pH), broth containing between 2-5 mg/L of Berenil effectively limited the growth of various strains of E.coli in the first 24 hours by as much as 77% compared to the controls. However the growth of Enterococcus faecalis, Listeria monocytogenes and Staphylococcus aureus was only significantly suppressed when the concentration of Berenil in the starting broth exceeded 50 mg/L. In the next part of this study we examined the effects of adding Berenil in mid-log phase on the growth of different strains of E.coli only. Low concentrations of Berenil (5mg/mL) did not induce any change in the growth pattern of K12 or E.coli 10005. However Berenil was a potent bactericide for both strains of E.coli O157:H7 inducing a collapse in both bacterial populations 6-8 hours later. Further investigation showed that the collapse in the population was dependent on the potassium ion concentration and the pH of the broth, and that the bactericidal action of Berenil was probably due the uncoupling of the membrane ion gradient in both strains of E.coli O157:H7. In the final part of this study we examined the effects of Berenil on the extracellular and intracellular peroxidase activity of E.coli O157:H7 as well as its influence on extracellular protease activity. It was found that the extracellular peroxidase activity of E.coli O157:H7 was reduced in a concentration dependent manner by Berenil. When compared to glucose, Berenil was 3-4 orders of magnitude more effective in reducing the extracellular peroxidase activity of E.coli O157:H7, however Berenil did not inhibit peroxidase activity directly only the secretion of peroxidases. Lastly a screen of 21 p-NA substrates was used to assess the extracellular protease/proteolytic activity of E.coli O157:H7. It was found that E.coli O157 and K12 in glucose (5%) assay media showed enhanced activity for 2 substrates, H-Asp-pNA and Ac-Asp-pNA. Activity was further enhanced (factor of 2-3) for H-Asp-pNA when Berenil was added to the medium.

Assessment of the PGRN and MAPT mutations in young(<65 years)Korean dementia patients

Lingyan Shen Gachon University Graduate School 2014 국내석사

Frontotemporal dementia (FTD) is the second most common form of early-onset (<65 years of age) neurodegenerative disease after Alzheimer’s disease (AD). At least seven genes have been reported as responsible risk factors for FTD, of which PGRN and MAPT were identified as the main disease-causing genes with 50% of the frequency for FTD, especially for familial FTD. To date, only a few studies of the two genes were performed in Asia, especially in Korea. The aim of this study is to investigate PGRN and MAPT mutations in young Korean dementia patients, and to explore the genetic and clinical features of various neurodegenerative disorders in Korea. PGRN and MAPT were analyzed by direct sequencing for 110 young Korean patients with early onset dementia (89EOAD and 21FTD), followed by single strand conformation polymorphism (SSCP) analyses, web-based splice sites prediction and/or protein function prediction, and in silico prediction and protein modeling. One novel deletion mutation (c.IVS8+23_+26delTGGG), one novel missense mutation (c.1767G>T, Leu589Phe; Leu434Phe and/or Leu409Phe), one novel intronic polymorphism (c.IVS12+56G>C), one common mutation which might be novel in Asia/Korea (rs113219200) and four common mutations (rs9897526, rs850713, rs5848 and rs25646; Asp108) were detected in the PGRN gene. But only two known silent mutations (rs115381139; Asn727 and rs11568305; Pro587) were identified in the MAPT gene. In addition, seven new types of co-existed SNPs in the PGRN gene were found in nine patients. Unfortunately, SSCP failed because of the huge size of PCR products. Web-based splice sites predictors showed the novel deletion mutation (c.IVS8+23_+26delTGGG) might cause disturbances in the splicing of PGRN transcript, since it is located near one splice site. Protein function prediction for the functional effect in GRNs (3 isoforms) caused by the novel missense mutation (c.1767G>T, Leu589Phe; Leu434Phe and/or Leu409Phe) showed possibly damaging (0.878) or benign (0.176), probably damaging (0.998) or probably damaging (0.933), and probably damaging (0.958) or possibly damaging (0.536) results based on the structures of GRN1, GRN2 and GRN3, respectively. In silico models showed the novel missense mutation (c.1767G>T, Leu589Phe; Leu434Phe and/or Leu409Phe) was located in the random coil structure at the C-terminal of GRNs (3 isoforms). Since phenylalanine and leucine are nonpolar and hydrophobic animo acids, random coil structure at the C-ternimal of GRNs (3 isoforms) might be changed by this novel point mutation (c.1767G>T, Leu589Phe; Leu434Phe and/or Leu409Phe). This work provided new insight into study for FTD in Asia.

AB208: A Chimeric Ligand With Superior Differentiation Capacity to Produce Insulin

Chia Yun Tseng Gachon university 2013 국내석사

Adult stem cells have great potential application for the therapeutic approach in the field of regenerative medicine. Due to their easy access and ethical issue, adipose‐derived stem cell has become increasingly popular. Many studies have stated Activin A, one of the TGF‐ β family, played a critical role for stem cell differentation to insulin producing cell. AB208, achimeric ligand of Activin‐bA and BMP‐2, exhibits the refolding characteristics of BMP‐2 while possessing Activin‐like signaling attributes. We isolated adipose‐derived stem cell from canine fat tissues and used AB208 to replace Activin A in two step induction protocol based on DMEM/F12, N2 supplement , B27 supplement, FGF2, nicotinamide, exdenin‐4, HGF and valproic acid for a duration of 16 days. After differentiation, the expression of genes related to β‐cell differentiation, INS, PDX‐1, and NGN‐3, was determined by real‐time PCR. The cASCs incubated with AB208 showed 2.5 times higher gene expression than these incubated with Activin A. The result of glucose challenage test of differentiation cells shows that cASCs differentiated with AB208 secreat more insulin than these differentiated with Activin A. These initail data show that AB208 has differentiation capacity to produce insulin and could be used as a therapeutic approach to treat Diabetes in the future. Key word: canine ASC, insulin producing cell/ cluster, Activin A , Type 1 Diabetes, pancreatic islet

Optical birefringence induced by surface plasmon resonance in optical fiber with bi-metallic thin film deposition

Nguyen Tan Tai Gachon University Graduate School 2012 국내석사

Surface Plasmon Resonance (SPR) technique with optical fiber technology has enormous advancements in sensing of various physical, chemistry and biochemistry parameter. In this work, we present the optical birefringence properties of the optical fiber cladding that exist as an evanescent field where the refractive index of the analyte solution is applied for optical sensor aspiration. To enhance the performance of the sensor, we have investigated different thickness of TiO2 coating, and bimetallic thin film alloy combinations. In addition, we describe a balanced homodyne detection method for the intensity difference change between the s and p polarization lights in the surface plasmon resonance sensing systems is strongly determined by the refractive index of the test sample. The proposed method can measure very small change of refractive index of glycerol solution, which 4.37x10-8 refractive index units (RIU). This method is a simple optical setup, easily to operate in the real time and low cost. Therefore, due to the experiment set up, our method can also eliminate the surrounding noise. Keywords: Surface Plasmon Resonance, Evanescent wave, Homodyne interferometry, Balanced detector.