303 research outputs found
Optimasi Kit Zeesan SARS-COV-2 Test Kit untuk Pengujian Deteksi DNA Porcine pada Produk Olahan Daging dan Produk Olahan Kompleks: Optimization of Zeesan SARS-COV-2 Test Kit for Porcine DNA Detection Tests on Processed Meat Products and Complex Processed Product
No full textVarious brands of covid 19 RNA detection kits are widely used by covid 19 testing laboratories. One of these kits is the Zeesan SARS COV-2 Test Kit. The Zeesan kit cannot be used to detect porcine DNA. There has been no official research on the use of optimized covid 19 detection kits for porcine DNA detection, thus the optimization of the Zeesan SARS COV-2 Test Kit used to detect porcine DNA is a new innovation in molecular testing. The purpose of this study was to find the right combination between the Zeesan SARS COV-2 Test Kit detection kit and the porcine primer-probe. Then look for the appropriate temperature setting for the amplification process for porcine DNA testing. Analysis using Liferiver brand real-time PCR with hydrolysis probe method. The study was conducted by running 3 types of positive porcine samples with 3 different concentrations. The research sample used sausage, ham, and noodles. The results showed that the combination detection kit of Zeesan SARS COV-2 Test Kit-primary probe porcine successfully detected porcine DNA in 3 positive samples of porcine with a concentration of 10 ng and 5 ng. Meanwhile, at a concentration of 1 ng, it could only be detected in 2 positive samples. In addition, this study showed a relatively faster detection time of porcine DNA than several previous studies
Development of a Soluble KIT Electrochemical Aptasensor for Cancer Theranostics
No full textAn electrochemical sensor based on a conformation-changing aptamer is reported to detect soluble KIT, a cancer biomarker, in human serum. The sensor was fabricated with a ferrocene-labeled aptamer (K(d) < 5 nM) conjugated to a gold electrode. Quantitative KIT detection was achieved using electrochemical impedance spectroscopy (EIS) and square-wave voltammetry (SWV). EIS was used to optimize experimental parameters such as the aptamer-to-spacer ratio, aptamer immobilization time, pH, and KIT incubation time, and the sensor surface was characterized using voltammetry. The assay specificity was demonstrated using interfering species and exhibited high specificity toward the target protein. The aptasensor showed a wide dynamic range, 10 pg/mL–100 ng/mL in buffer, with a 1.15 pg/mL limit of detection. The sensor also has a linear response to KIT spiked in human serum and successfully detected KIT in cancer-cell-conditioned media. The proposed aptasensor has applications as a continuous or intermittent approach for cancer therapy monitoring and diagnostics (theranostics)
Existence of Solutions to Wasserstein Gradient Flows and Their Long Time Asymptotic Behaviors
No full textNg, Wing Kit.Thesis M.Phil. Chinese University of Hong Kong 2016.Includes bibliographical references (leaves ).Abstracts also in Chinese.Title from PDF title page (viewed on …)
Prevalence of KIT D816V in anaphylaxis or systemic mast cell activation
No full text409418Background: The KIT D816V mutation is a hallmark of clonal mast cell disease (cMCD) including systemic mastocytosis. cMCD diagnosis is frequently delayed because of variable symptoms until confirmatory bone marrow biopsy is performed. Screening techniques for KIT D816V variant in peripheral blood (PB) may facilitate diagnosis. Prevalence of KIT D816V in patients with anaphylaxis or mast cell activation (MCA) symptoms is unknown.
Objective: We sought to determine the prevalence of KIT D816V in PB of patients with anaphylaxis or systemic MCA symptoms.
Methods: The PROSPECTOR trial (NCT04811365) included patients with anaphylaxis or systemic MCA symptoms who had (1) moderate to severe anaphylaxis to Hymenoptera sting, (2) 20% + 2 ng/mL increase in tryptase level over the baseline level during moderate to severe anaphylaxis with cardiovascular involvement, and/or (3) involvement of both the cardiovascular system and 1 or more other organ systems with basal serum tryptase (BST) levels greater than or equal to 8 ng/mL. KIT D816V in PB, hereditary α-tryptasemia (HaT), and BST levels were centrally evaluated.
Results: Of the 381 enrolled patients, 179, 76, and 203 were in groups 1, 2, and/or 3, respectively. Fifteen patients (4%) had detectable KIT D816V in PB; 12 of 15 (80%) had BST levels less than 20 ng/mL. Most patients with KIT D816V (11 of 15 [73%]) had Ring-Messmer grade III/IV anaphylaxis. Fourteen additional patients with BST levels greater than 11.4 ng/mL, no HaT, and local follow-up were diagnosed with cMCD, totaling 29 of 381 patients (8%) with cMCD in the PROSPECTOR trial. The overall prevalence of HaT was 36% (138 of 381).
Conclusions: The PROSPECTOR trial demonstrated a meaningful KIT D816V prevalence in patients with anaphylaxis or systemic MCA symptoms; more frequent and sensitive screening for KIT D816V is needed in this patient population.157
Differential early gene expression in HBV X protein (HBx)-mediated hepatocarcinogenesis.
No full textby Ray, Kit Ng.Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.Includes bibliographical references (leaves 112-121).Abstracts in English and Chinese.Abstract --- p.iAcknowledgments --- p.ivAbbreviations --- p.xList of Figures --- p.xiiList of Tables --- p.xivChapter Chapter 1 --- IntroductionChapter 1.1 --- Hepatitis B Virus (HBV) --- p.1Chapter 1.2 --- Hepatitis B Virus X Protein (HBx) --- p.5Chapter 1.2.1 --- The Genomic Structure of HBx --- p.5Chapter 1.2.2 --- The HBx Protein Structure --- p.6Chapter 1.2.3 --- Subcellular Localization of HBx --- p.7Chapter 1.2.4 --- Possible Functions of HBx --- p.8Chapter 1.3 --- Etiology of Hepatocellular Carcinoma (HCC) --- p.12Chapter 1.4 --- Relationship between HCC and HBx --- p.13Chapter 1.5 --- Aims of Study --- p.14Chapter 1.6 --- The Basis of Tet-On System --- p.15Chapter 1.7 --- The Basis of DNA Microarray --- p.18Chapter 1.8 --- The Basis of Two-Dimensional Electrophoresis --- p.20Chapter Chapter 2 --- Materials and MethodsChapter 2.1 --- Construction of a Tet-On HBx Expressing Cell Model --- p.22Chapter 2.1.1 --- Cloning of HBx Gene into pTRE2 Vector --- p.22Chapter 2.1.1.1 --- PCR of HBx Gene --- p.22Chapter 2.1.1.2 --- Purification of the PCR Product --- p.23Chapter 2.1.1.3 --- Restriction Enzyme Digestion --- p.23Chapter 2.1.1.4 --- Ligation of HBx into pTRE Vector --- p.24Chapter 2.1.1.5 --- Transformation of the Ligation Product into Competent Cells --- p.24Chapter 2.1.2 --- Preparation of the Plasmid DNA --- p.24Chapter 2.1.2.1 --- DNA Sequencing of the Cloned Plasmid DNA --- p.25Chapter 2.1.3 --- Cell Culture of AML12 Cell Line --- p.26Chapter 2.1.4 --- Transfection of pTet-On Vector into AML12 Cells --- p.26Chapter 2.1.5 --- Selection of the Transfected AML12 Cells by G418 --- p.27Chapter 2.1.6 --- Single Clone Isolation --- p.27Chapter 2.1.6.1 --- Luciferase Assay for Selection of Highly Inducible Clones --- p.28Chapter 2.1.7 --- Second Transfection of pTRE-HBx Plasmid --- p.28Chapter 2.1.8 --- Selection of the Transfected Cells by Hygromycin --- p.29Chapter 2.1.9 --- Second Single Clone Isolation --- p.29Chapter 2.1.10 --- Total RNA Isolation --- p.29Chapter 2.1.11 --- DNase I Digestion --- p.30Chapter 2.1.12 --- First-Strand cDNA Synthesis --- p.31Chapter 2.1.13 --- RT-PCR of HBx Gene --- p.31Chapter 2.1.14 --- Northern Blotting --- p.32Chapter 2.1.15 --- Preparation of the Probe --- p.33Chapter 2.1.16 --- Northern Blot Hybridization --- p.33Chapter 2.1.17 --- 3H-Thymidine Incorporation Assay --- p.34Chapter 2.1.18 --- Analysis of Cell Cycle by Flow Cytometry --- p.35Chapter 2.2 --- Microarray Analysis of Differential Gene Expression upon HBx Induction --- p.35Chapter 2.2.1 --- Sample Preparation for Microarray Analysis --- p.35Chapter 2.2.2 --- Probe Labelling --- p.36Chapter 2.2.3 --- Microarray Hybridization --- p.37Chapter 2.2.4 --- RT-PCR of the Candidate Genes --- p.38Chapter 2.2.5 --- Northern Blot Analysis of the Candidate Genes --- p.39Chapter 2.3 --- Two-Dimensional (2D) Gel Electrophoretic Analysis --- p.40Chapter 2.3.1 --- Protein Sample Preparation for 2D Gel Electrophoresis --- p.40Chapter 2.3.2 --- First-Dimension Isoelectric Focusing (IEF) --- p.40Chapter 2.3.3 --- Second-Dimension SDS-PAGE --- p.41Chapter 2.3.4 --- Silver Stain of 2D Gel --- p.42Chapter 2.3.5 --- Mass Spectroscopic Analysis --- p.43Chapter 2.4 --- Subcellular Localization of HBx --- p.44Chapter 2.4.1 --- Cloning of HBx into Green Fluorescent Protein (GFP) Expression Vector --- p.44Chapter 2.4.2 --- Transfection of GFP-HBx --- p.44Chapter 2.4.3 --- Propidium Iodide (PI) Staining --- p.45Chapter 2.4.4 --- Mitochondria Staining --- p.45Chapter 2.4.5 --- Subcellular Localization Study using Epi-Fluorescent Microscopy --- p.45Chapter 2.5 --- Analysis of Mitochondrial Transmembrane Potential --- p.46Chapter Chapter 3 --- ResultsChapter 3.1 --- Construction of Tet-On AML12 Cell Line of HBx Gene --- p.47Chapter 3.2 --- Characterization of the HBx-Expressing Cell Model --- p.53Chapter 3.2.1 --- 3H-Thymidine Proliferation Assay --- p.53Chapter 3.2.2 --- Cell Cycle Analysis --- p.55Chapter 3.3 --- Microarray Analysis of Differential Gene Expression Pattern upon HBx Induction --- p.57Chapter 3.4 --- Northern Blot Analysis and RT-PCR of the Candidate Genes --- p.65Chapter 3.5 --- Differential Protein Expression Pattern under HBx Induction --- p.70Chapter 3.6 --- Subcellular Localization of HBx --- p.77Chapter 3.7 --- Analysis of Mitochondrial Transmembrane Potential --- p.83Chapter Chapter 4 --- DiscussionChapter 4.1 --- Conditional HBx-Expressing Cell Model --- p.84Chapter 4.2 --- The Effects of HBx in Clone X18 --- p.86Chapter 4.2.1 --- Proliferative Effect of HBx --- p.86Chapter 4.2.2 --- Deregulation of G2/M Checkpoint by HBx --- p.86Chapter 4.3 --- Early Differential Gene Expression due to HBx Induction --- p.88Chapter 4.4 --- The Relationship of the Potential Candidate Genes and Cancer Development --- p.90Chapter 4.5 --- The Protein Expression Pattern due to HBx Induction --- p.93Chapter 4.6 --- The Subcellular Localization of HBx --- p.96Chapter 4.7 --- The Possible Involvement of HBx in Mitochondrial Transmembrane Potential --- p.98Chapter 4.8 --- Conclusions --- p.101Chapter 4.9 --- Future Prospects --- p.104Appendix --- p.107References --- p.11
CUHK electronic theses & dissertations collection
No full textTsang, Kit Man.Thesis M.Phil. Chinese University of Hong Kong 2014.Includes bibliographical references (leaves 125-137).Abstracts also in Chinese.Title from PDF title page (viewed on 04, November, 2016)
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Design and Analysis for the Sphinx-NG CubeSat
No full textThis project presents the mechanical, orbital, structural design and analysis for a three-unit Cube Satellite (CubeSat) with the SphinX-NG instrument as payload. The goal of the mission is to place the CubeSat on a near-polar, sun synchronous orbit where it will perform X-ray spectroscopy. The design complies with the Poly-Picosatellite Orbital Deployer (P-POD) and mission requirements. The mechanical design is performed using SolidWorks. Orbital decay analysis using the Systems Tool Kit shows that a 600 km altitude provides a compliant lifetime. Random vibration analysis performed with ANSYS under the maximum expected loads shows compliance. COMSOL analysis of the magnetic fields induced by the onboard magnetic torquers indicates the need for shielding of the onboard magnetometer
Abstract 5526: Tumor suppressive role of <i>BMI-1</i> through inhibition of JAK-STAT signaling in leukemia
No full textAbstract
BMI-1, which is one of the core components of polycomb repressive complex 1, is frequently found deregulated in patients with hematological disorders. In last decades, researchers concordantly agree that BMI-1 mediates tumorigenesis of leukemia stem cells through p16INK4A leukemogenic pathway. However, accumulating evidences contradict the idea that BMI-1 solely plays an oncogenic role in tumorigenesis. It has been shown BMI-1 depletion favors the development of myelofibrosis in mice; whereas high BMI-1 expression suppressed colony forming ability of MLL-ENL-transformed bone marrow and correlated with higher survival in some cancers. In this study, we hypothesized that BMI-1 plays a tumor suppressive role, which is independent of the regulation of INK4A-ARF locus, in human leukemia. BMI-1 was over-expressed in a panel of myeloid and lymphoid lineage leukemia cells, including HL-60, MV4-11, MonoMac-6, SEM, Nalm-20 and RS4;11. We observed no deregulation of p16INK4A and p14ARF genes by BMI-1, suggesting the regulation of INK4A-ARF locus is independent of BMI-1 modulation in leukemia cells. Nevertheless, over-expression of BMI-1 resulted in significant reduction of leukemia cell proliferation. It is noted that constitutively active JAK-STAT signaling pathway is crucial to leukemia cell survival. By modulation of BMI-1 level, we demonstrated suppression of the activated JAK-STAT signaling pathway in most of the leukemia cell lines with the exception of MonoMac-6 and RS4;11. This is in agreement with the high sensitivity to ruxolitinib, a JAK-STAT inhibitor, in all the tested leukemia cell lines except RS4;11. Importantly, we showed that BMI-1 over-expression could sensitize RS4;11 cells to reduce cell proliferation under ruxolitinib treatment. These results suggest that higher efficacy of ruxolitinib treatment could be achieved under a condition of high cellular level of BMI-1. We further demonstrated that ruxolitinib treatment was more effective in a cohort of AML patient samples (n = 25) with a context of higher BMI-1 expression (p &lt; 0.05). Altogether, our results suggest that BMI-1 functions as a tumor suppressor gene via inhibition of the JAK-STAT signaling pathway. The endogenous level of BMI-1 could be served as an indicator for the effective treatment of JAK-STAT-dependent leukemia cells using ruxolitinib.
Citation Format: Yuk Man Lam, Stephen Sze Yuen Lam, Anskar Yu Hung Leung, Ray Kit Ng. Tumor suppressive role of BMI-1 through inhibition of JAK-STAT signaling in leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5526. doi:10.1158/1538-7445.AM2017-5526</jats:p
Application of an X-ray microscopy technique to evaluate tissue-engineered bone-scaffold constructs
No full text10.1016/S0928-4931(02)00007-3Materials Science and Engineering C201-29-1
Teknik Isolasi DNA dari Daging Ikan Salmon (Oncorhynchus masou) Tuna (Thunnus obesus) dan Tongkol (Euthynnus affinis) Menggunakan Metode Spin Column
No full textPenelitian ini bertujuan untuk mengevaluasi efektivitas metode spin column dalam isolasi DNA dari jaringan daging ikan Salmon (Oncorhynchus masou), Tuna (Thunnus obesus), dan Tongkol (Euthynnus affinis), tiga spesies bernilai ekonomi tinggi di sektor perikanan Indonesia. Isolasi DNA dilakukan menggunakan metode spin column dengan kit Tianamp, melalui tahapan lisis jaringan daging ikan, presipitasi DNA, pencucian kontaminan, dan elusi akhir, kemudian dianalisis kuantitas dan kemurniannya menggunakan spektrofotometri pada panjang gelombang 260/280 nm. Hasil menunjukkan konsentrasi DNA berkisar 50.8–183.3 ng/μL (rata-rata 109.3 ng/μL), namun kemurnian DNA (A260/A280) berada pada kisaran 1.19–1.83 (rata-rata 1.46), mengindikasikan kontaminasi protein/RNA. Ikan Salmon menunjukkan kemurnian tertinggi (1.83), sedangkan Tongkol memiliki konsentrasi tertinggi (183.3 ng/μL) tetapi kemurnian terendah (1.19). Diduga faktor pemanasan dan inhibitor sebagai penyebab rendahnya kemurnian DNA pada penelitian ini. Rekomendasi optimasi meliputi modifikasi suhu inkubasi, penggunaan fenol-kloroform, dan pengelolaan pasca-panen. Penelitian ini memberikan kontribusi dalam pengembangan teknik isolasi DNA untuk aplikasi biologi molekuler dan industri perikanan
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