Dissertationen zum Thema „Metalloproteinases Inhibitors Therapeutic use“
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1
Tang, Wing-yan, und 鄧詠欣. „Curcumin inhibits cancer cells migration and invasion of tongue carcinoma through down-regulation of matrix metalloproteinase-10“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B47869768.
Annotation:
Squamous cell carcinoma of the tongue has a low survival rate, with cure rate reduced by half if cervical lymph node metastasis is present. Standard treatment regimen includes surgical resection of the tumor, radiotherapy and chemotherapy. These treatment modalities, however, can result in irreversible side effects including loss of form and function of the tongue. So far, there is no efficient treatment regime targeting migration and invasion of tongue carcinoma.
Curcumin is a natural polyphenol extracted from Curcuma longa. Recent studies indicated that curcumin is a potential anti-cancer agent. The anticancer effects have been demonstrated in numerous cancers including lung cancer, liver cancer, breast cancer, prostate cancer and melanoma. In head and neck cancers, the number studies is limited and its inhibitory effects in migration and invasion is rarely explored.
To explore the global expression changes in tongue cancer, we used microarray to evaluate the genes responsive to curcumin treatment and focused on genes related to migration and invasion of tongue cancer cell line HN21B. The genes down-regulated by curcumin were validated in HN21B and two other tongue cancer cell line CAL27 and HN96 using qRT-PCR, Western blotting and immunostaining. The identified genes were quantified in tongue carcinoma tissues to examine whether it was up-regulated in human tongue tissues. Scratch wound assay and radial-migration assay were used to assess the degree of inhibition on migration. Adhesion and invasion assays were also performed to assess the adhesion and invasion ability.
Transcriptomic analyses showed that MMP-10 was 2.36 fold down-regulated in HN21B in response to curcumin. Curcumin treatment resulted in down-regulation of MMP-10 gene in all the 3 tongue carcinoma cell lines at mRNA and protein levels. Out of 24 tongue carcinoma cases, 55% tumor tissue had obvious up-regulation of MMP-10 expression in comparison with the normal counterpart. Adhesion, migration and invasion ability of tongue carcinoma cell lines was significantly reduced upon IC50 of curcumin treatment in all TSCC cell lines.
In conclusion, our results indicated that curcumin could reduce migration, adhesion and invasion in tongue carcinoma cells partly through reducing MMP-10 expression. Further investigations are warranted to explore the potential therapeutic use of curcumin to inhibit migration and invasion of tongue carcinoma cells.published_or_final_version
Surgery
Master
Master of Philosophy
2
Roach, Denise Margaret. „Upregulation of matrix metalloproteinases -2 and -9 and type IV collagen degradation in skeletal muscle reperfusion injury“. Title page, contents and abstract only, 2002. http://web4.library.adelaide.edu.au/theses/09MD/09mdr6281.pdf.
Annotation:
Includes bibliographical references (leaves 292-352) Determines the role of matrix metalloproteinases, MMP-2 and MMP-9 in reperfusion injury following skeletal muscle ischaemia; and, whether inhibition of MMPs by doxycycline protects against tissue damage.
3
Peng, Xiaofang, und 彭晓芳. „Naturally occurring inhibitors against the formation of advanced glycation endproducts“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2010. http://hub.hku.hk/bib/B44892706.
4
Sun, Wentao, und 孙文韬. „The effects of Panax notoginseng extracts and its components on TNF-alpha induced MMP-9 expression and activity“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2014. http://hdl.handle.net/10722/207686.
Annotation:
Matrix metalloproteinase (MMP) induced extra cellular matrix (ECM) degradation is a crucial process involved in the development of many chronic inflammatory diseases, including cardiac remodeling and cancer metastasis. In cardiac remodeling, the presence of pathological stimuli leads to elevated MMP-9 expression and impairment of cardiac performance, which subsequently develops into heart failure. While in tumorgenesis, MMP-9 has been found to play key roles in metastasis, as it can break physical barriers for the tumor. Therefore, searching for agents targeting MMP-9 is a new direction for the treatment of cardiac remodeling and cancer metastasis.
Chinese herbal medicine is becoming increasingly used worldwide in recent decades. In the past twenty years, as many highly selective and sensitive bioassays were introduced into the bioactive compounds screening from herbal medicine, more than one hundred new drug candidates have been identified. Therefore, herbal medicine is a potential source of bioactive compounds. Panax notoginseng (PNG) is one of the most common traditional Chinese medicines to treat cardiovascular diseases, and it was also reported to have anti-cancer effect. We hypothesized that it contains bioactive compounds that could inhibit MMP-9 activity in cardiomyocytes and cancer cells.
In order to examine the effect of PNG on cardiac remodeling and cancer metastasis, we employed TNF-α induced MMP-9 in H9c2 cell (a rat cardiomyocyte) and HepG-2 cell (a human hepatoma cell) as an in vitro assay, respectively. PNG was first extracted by four different extraction methods according to the polarity of the solvent. The most effective fraction in suppressing MMP-9 activity in TNF-α induced H9c2 cell was chosen for further separation by silica gel column chromatography and high performance liquid chromatography (HPLC) until a single compound was isolated. According to the result of spectroscopic analysis by NMR, the compound was identified as ginsenoside Rb1. For the bioactivity assays, real-time quantitative polymerase chain reaction (QPCR) and Enzyme-linked immunosorbent assay (ELISA) were used to measure the mRNA and protein expression of MMP-9, respectively. We also examined the MMP-9 activity by gelatin zymography. The results showed that both of the PNG extract obtained from 10% ethanol extraction method (PNG-3) and purified Compound P (ginsenoside Rb1) showed significant inhibitory effect on MMP-9 expression and activity in H9c2 cells and HepG-2 cells.
We further examined the molecular mechanisms of the inhibitory effect of PNG-3. H9c2 and HepG-2 cells were pretreated with different kinase inhibitors followed by the activation by TNF-α. The results showed the protein kinase R (PKR) inhibitor could inhibit TNF-α induced MMP-9 in both of the two cell lines. Furthermore, the results of Western blot showed the PNG-3 suppressed the phosphorylation of eIF-2α which is a down-stream effector of PKR in TNF-α stimulated H9c2 and HepG-2 cells, respectively. Therefore, PNG-3 may act through PKR to regulate TNF-α induced MMP-9 activity.
In summary, bioactivity guided fractionation is an effective way of isolating bioactive compounds from medicinal herbs. In addition, PNG containing ginsenoside Rb1 may be a potential candidate of MMP-9 inhibition for the treatment of cardiac remodeling and cancer metastasis.published_or_final_version
Paediatrics and Adolescent Medicine
Master
Master of Philosophy
5
Chan, Hoi-ching, und 陳凱靜. „Heat shock protein 90 inhibitor 17-AAG potentiates anticancer activityof bortezomib in NK cell malignancies“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46632025.
6
He, Ru, und 何茹. „Effectiveness and toxicity of aromatase inhabitors [i.e. inhibitors] in adjuvant therapy for hormone receptor positive postmenopausalbreast cancer: a meta-analysis“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46936026.
7
Gu, Baoying. „Selective increase of neuronal cyclooxygenase-2 (COX-2) expression in vulnerable brain regions of rats with experimental Wernicke's encephalopathy : effects of nimesulide“. Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=112627.
Annotation:
Wernicke's encephalopathy is a neuropsychiatric disorder resulting from thiamine deficiency (TD) and is characterized by neuronal loss, astrocytic proliferation and microglial activation. Cyclooxygenases (COX) are enzymes which catalyze the first step in the synthesis of prostanoids. COX-1 is expressed constitutively and COX-2 is the inducible isoform. Groups of TD rats and pair-fed controls were killed at presymptomatic and symptomatic stages of encephalopathy. Cresyl violet and NeuN staining showed decreased numbers of neuronal cells in vulnerable regions (medial thalamus and inferior colliculus) but not in a spared region (frontal cortex). Numbers of GFAP-positive and OX-42-positive cells were increased at symptomatic stage of encephalopathy. Expression of COX-2 mRNA and neuronal COX-2 immunoreactivity were selectively increased in vulnerable regions of TD rats at symptomatic stages of encephalopathy. Nimesulide, a highly selective COX-2 inhibitor, lowered PGE2 levels and precipitated the progression of encephalopathy suggesting that COX-2 in this model is conferring neuroprotection.
8
Ho, Kwun-wai, und 何冠威. „Angiotensin converting enzyme inhibitor alone or in combination with angiotensin II type I receptor blocker in patients with chronicproteinuric nephropathies: a systemic reviewof clinical trials“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2005. http://hub.hku.hk/bib/B45010687.
9
Scrivens, Paul James. „Regulation and chemotherapeutic targeting of human Cdc25A phosphatase“. Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=103293.
Annotation:
The Cdc25 phosphatases are highly conserved from yeast through humans and play pivotal roles in regulating the activities of cyclin-dependent kinases (Cdks). Cdc25A is one of three human Cdc25 family members, and has previously been shown to be overexpressed in numerous cancers and to transform rodent fibroblasts. Cdc25A therefore represents a rational target for chemotherapeutic development. Further, a thorough understanding of its biology and regulation in normal and transformed cells may facilitate the development of strategies to specifically interfere with the proliferation of cancerous cells. In this work I describe experiments which demonstrate that bisperoxoVanadium compounds, and specifically bpV(Me2Phen), inhibit Cdc25A phosphatase in vitro and in vivo. Further, these compounds cause cell-cycle arrest, are cytotoxic to cancer cells, and slow the growth of tumours in mouse models. With respect to the fundamental biology of Cdc25A, I have identified a sequence element (NLS) responsible for nuclear localization of Cdc25A phosphatase. An analysis of this sequence demonstrated high conservation of flanking phosphoacceptor sites, notably Serine 292. S292 was predicted to be a consensus PKA or CamKII substrate. Using site-directed mutagenesis I have shown that S292 is the sole site of PKA phosphorylation in vitro. The functional importance of S292 phosphorylation was investigated via transfections of phospho-mimetic mutants of S292 (S292E) expressed as GFP-fusion proteins; these studies indicate that S292 phosphorylation may promote nuclear localization. Studies by other groups have indicated that S292 is a phosphorylation site for inhibitory kinases, namely Chk1 and Chk2 (4). I generated a phospho-specific antibody to this site and demonstrate by immunofluorescence and western blotting an unexpected pattern of S292 phosphorylation associated with nuclear bodies and the mitotic apparatus. I provide evidence to suggest that these sites represent local fine-tuning of Cdc25A, allowing Cdk activity to be controlled at the level of specific subcellular structures. These studies highlight the complexity of Cdc25 regulation and indicate a previously unappreciated degree of control of their activity such that these enzymes exist in multiple discrete pools within a given cell.
10
Wong, Carmen, und 黃嘉敏. „A systematic review of the drug sorafenib in extending survival time in patients with hepatocellular carcinoma“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46942865.
11
Page, Simon Matthew. „Ruthenium anticancer complexes : a targeted approach to enzyme inhibition“. Thesis, University of Cambridge, 2013. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.608027.
12
Brice, Edmund Andrew William. „Rat angiotensin-converting enzyme : tissue specific expression during pharmacological inhibition“. Doctoral thesis, University of Cape Town, 1995. http://hdl.handle.net/11427/27042.
Annotation:
The renin-angiotensin system plays a central role in the maintenance of blood pressure. Angiotensin II, the main effector of this system, results from the action of angiotensin-converting enzyme (ACE) on angiotensin I. Angiotensin II, maintains vasomotor tone via its vasoconstrictor action, and also increases salt and water retention by stimulating the release of aldosterone. ACE inhibitors, such as captopril, enalapril and lisinopril, are highly effective in the treatment of hypertension and congestive cardiac failure. Previous studies have suggested that angiotensin converting enzyme (ACE) production may be enhanced during pharmacological inhibition of the enzyme. Little is known, however about the mechanism of this induction. After demonstrating increases in circulating ACE protein in cardiac failure patients receiving the ACE inhibitor captopril, a rat model was used to study this effect. A sensitive enzyme linked immunosorbent assay for rat ACE was developed and a partial cDNA for rat ACE cloned to enable examination of ACE mRNA and protein expression during enzyme inhibition with enalapril. Rat lung ACE mRNA increased by 156% (p<0.05) and ACE protein doubled within 3 hours of administering a single dose of enalapril. Testicular ACE mRNA also increased by 300% (p<0.05) within 2 hours and returned to pretreatment levels by 6 hours. The angiotensin II antagonist saralasin similarly caused a significant (p<0.0001) 800% enhancement of mRNA expression. Aldosterone pretreatment of rats prior to enalapril administration was found to abolish this mRNA induction. These findings indicate that increased ACE expression during inhibition results from reduced levels of angiotensin II with consequent reduced stimulation of the angiotensin 11 receptor and its effects, such as aldosterone release. This suggests that ACE levels are regulated by a negative feedback loop involving the distal components of the renin-angiotensin system, namely angiotensin II and aldosterone. In situ hybridisation and immunohistochemical techniques were employed to localise the site of this inductive response in rat tissue sections. It was found that lung macrophages were markedly induced to produce ACE, as was ACE in seminiferous tubules. ACE induction was also noted in the expected sites of renal tubular epithelium and glomerular tissue. Interestingly, ACE expression was also enhanced in cardiac valves. In these studies it has been conclusively demonstrated that new ACE expression is induced by enzyme inhibitor therapy. A variety of techniques have been developed that will allow futher study of ACE in rat tissues.
13
Faridi, Nazlie. „Amphetamine-induced dopamine release in treatment-naïve men with ADHD : a PET[¹¹C]raclopride study“. Thesis, McGill University, 2008. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=111569.
Annotation:
Attention deficit hyperactivity disorder (ADHD) affects up to 10% of school-aged children and half as many adults. The core features of impulsivity, hyperactivity, and inattentiveness commonly give rise to academic underachievement, poor social relationships, and increased risk for mood and anxiety disorders. Although the relevant neurobiological mechanisms remain poorly understood, altered mesocorticolimbic dopamine (DA) transmission has been proposed. The aim of the present study was to compare striatal DA function in treatment-naive adults with ADHD vs. age- and IQ-matched controls. Two PET/[11C]raclopride scans, one with placebo and one with d-amphetamine (d-AMP; 0.3 mg/kg, p.o.), were administered to five men with ADHD and five healthy male volunteers. The ADHD group differed from controls in demonstrating significant d-AMP-induced reductions in posterior caudate (p<0.05). These results may support a proposed model of reduced DA tone leading to increased phasic signaling in ADHD.
14
Blagojevic, Ana. „An interaction between statins and clopidogrel : a pharmacoepidemiology cohort study with survival time analysis“. Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=112383.
Annotation:
Clopidogrel is an antiplatelet drug prescribed to prevent stent thrombosis after a percutaneous coronary intervention (PCI). Previous evidence suggests that some widely prescribed statins may inhibit the antiplatelet effects of clopidogrel via competitive metabolism of its activating enzyme cytochrome P450 3A4 (CYP3A4).The objective was to investigate the possibility of an interaction post-PCI between statins and clopidogrel.
We carried out a population-based cohort study identifying 10,491 patients using clopidogrel post-PCI (2001-2004). The outcome was a composite of death of any cause, myocardial infarction, unstable angina, repeat revascularization, and cerebrovascular events. We found that co-prescription of CYP3A4-metabolized statins (hazard ratio (HR) 0.95, 95% confidence interval (CI) 0.79-1.15), or non-CYP3A4-metabolized statins (HR 0.82, 95% CI 0.63-1.07) with clopidogrel was not associated with increase in adverse outcomes.
We observed no evidence of interaction between clopidogrel and statins in a large population cohort of PCI patients, suggesting unlikelihood of an important interaction.
15
Maloney, Shawn C. „Histopathology of human age-related macular degeneration and the development of a novel animal model“. Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=112539.
Annotation:
Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly worldwide. Due to the inadequacy of current pharmacotherapies, novel molecular targets must be sought as potential therapeutic candidates. Furthermore, there is a need for more efficient and cost-effective animal models of this pathology in order to accelerate in vivo investigations.Our laboratory is in possession of human choroidal neovascular membranes which we examined for expression of cyclooxygenase (COX)-2. This expression was characterized in retinal pigment epithelial, vascular endothelial, and fibroblast cells and correlated with patient age. We also looked at the feasibility of creating a rabbit laser-injury model to adequately mimic human neovascular AMD.
Our results suggest that anti-COX-2 therapies may be beneficial to some patients with neovascular AMD. Moreover, there is strong potential for the development of clinically relevant choroidal neovascularization in rabbits using the laser-injury technique. This approach may yield a novel, cost-effective AMD model.
16
Hubbe, Michelle E. (Michelle Elzabet). „Evaluation of antioxidant and free radical scavenging activities of honeybush tea (Cyclopia)“. Thesis, Stellenbosch : Stellenbosch University, 2000. http://hdl.handle.net/10019.1/51749.
17
He, Hua, und 何華. „Anti-tumor mechanisms of cyclooxygenase inhibitors and a c-Jun-N-terminal kinase inhibitor in gastrointestinal cancers“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2004. http://hub.hku.hk/bib/B30075245.
18
He, Xi, und 何溪. „The impact of lopinavir/ritonavir (Kaletra) on blood lipids in HIV/AIDS antivirus treated naïve patients in China“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46936129.
19
Lu, Xiaofan, und 陆小凡. „Mechanism study of a small molecule F18 as a novel anti-HIV-1 non-nucleoside reverse transcriptase inhibitor“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B47246509.
Annotation:
Non-nucleoside reverse transcriptase inhibitor (NNRTI) is one of the key components of antiretroviral drug regimen against human immunodeficiency virus type-1 (HIV-1) replication. However, the low genetic barriers to drug-resistance or cross-resistance, side effects, as well as the unaffordable cost of NNRTIs compromise their clinical usage. Therefore, to develop novel NNRTIs with potent antiviral activity against HIV-1 becomes a major concern in the treatment and prevention of HIV/AIDS.
(+)-Calanolide A, which is a natural product initially extracted from the tropical rainforest tree Calophyllum lanigerum, was identified as an attractive NNRTI against HIV-1 despite virus strains containing drug-resistant K103N/Y181C mutations. In this study, a chemical library was constructed based on the three chiral carbon centers of (+)-Calanolide A. After screening the activity against HIVNL4-3 wild-type and several NNRTI-resistant pseudoviruses, a small molecule 10-chloromethyl-11- demethyl-12-oxo-calanolide A (F18) was identified as novel NNRTI with promising anti-HIV efficacy.
Further studies were performed to investigate the antiviral breadth, drug resistance profile and underlying mechanism of the action of F18. F18 consistently displayed a potent activity against primary HIV-1 isolates including various subtypes of M group, CRF01_AE, and laboratory-adapted drug-resistant viruses in PBMC based assay. Moreover, F18 displayed distinct profiles against 17 NNRTI-resistant pseudoviruses, with an excellent potency especially against one of the most prevalent strains with the Y181C mutation (EC50=1.0nM) in cell line based assay, which was in stark contrast from the extensively used NNRTIs nevirapine and efavirenz. F18-resistant viruses were induced by in vitro serial passages, and mutation L100I was appeared to be the dominant contributor to F18-resistance, further suggesting a binding motif different from nevirapine and efavirenz. The efficacy of F18 was non-antagonistic when used in combination with other antiretrovirals against both wild-type and drug-resistant viruses in infected PBMCs. Interestingly, F18 displayed a highly synergistic antiviral effect with nevirapine against nevirapine-resistant virus (Y181C). Furthermore, in silico docking analysis suggested that F18 may bind to the HIV-1 reverse transcriptase in a way different to other NNRTIs. For the potential as an anti-HIV-1 microbicide, F18 also showed the stable and rapid release, as well as the sustained antiviral activity against HIV-1 wild-type virus in a formulation temperature-sensitive acidic gel.
In summary, this study presents F18 as a new potential drug for clinical use and also underlies new mechanism-based design for future NNRTI.published_or_final_version
Microbiology
Doctoral
Doctor of Philosophy
20
Assadian, Sarah. „Rodent FDG-PET imaging for the pre-clinical assessment of novel glioma therapies“. Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=101836.
Annotation:
The rapid discovery of novel therapeutic agents, targeting the specific mechanism of cancer progression, invasion and angiogenesis, necessitates the development and validation of efficient techniques to assess the therapeutic efficacy of these drugs in vivo. Recently the development of dedicated PET scanners for the imaging of small animals, such as the microPET system (CTI Concorde R4), has allowed for the high-resolution functional and molecular imaging of murine and rodent models of disease. This study, investigates the ability of microPET imaging, using the 18F labelled 2-fluoro-2-deoxyglucose (FDG) PET tracer, to detect the therapeutic efficacy of novel targeted therapies in a rat model of glioma. This technique potentially allows for the rapid and high-throughput assessment of tumour response and evaluation of efficacy of such therapeutic agents in vivo at the pre-clinical stage and will, consequently, facilitate the translation of these novel drugs from the discovery to the clinical phases.La découverte accélérée de nouvelles molécules thérapeutiques qui ciblent lesmécanismes de progression du cancer tels que l'invasion et l'angiogenèse, nécessite lamise au point et la validation de techniques efficaces qui permettent d'évaluer l'efficacitéthérapeutique de ces agents in vivo. Le développement récent des scanners detomographie à émission de positron (TEP) dédiés à l'imagerie de petits animaux(microPET, CT! Concorde R4), permet aujourd'hui d'obtenir une image fonctionnelle etmoléculaire de haute résolution des modèles rongeurs. Cette étude s'intéresse au potentieldu 18F-2-fluoro-2-deoxyglucose (FDG) en utilisant l'imagerie microPET dansl'évaluation de l'efficacité de nouveaux agents thérapeutiques dans un modèle de gliomechez le. rat. Cette technique pourrait éventuellement mener à une évaluation rapide et àgrande échelle de la réponse tumorale, ainsi que la mesure de l'efficacité d'agentsthérapeutiques in vivo au stade d'étude préclinique. Globalement, cette étude a pour butde faciliter la transition entre la découverte de nouvelles molécules thérapeutiques et leursapplications cliniques.
21
Hiss, Donavon Charles. „Perturbation of glycoprotein expression and processing in multidrug resistant cells : modulation of drug transport and cytotoxicity by Tunicamycin“. Doctoral thesis, University of Cape Town, 1994. http://hdl.handle.net/11427/26338.
22
Davies, Richard. „Effect of selective COX-2 inhibitors on hepatic progenitor cells and the pathologies of experimental hepatocarcinogenesis“. University of Western Australia. School of Medicine and Pharmacology, 2007. http://theses.library.uwa.edu.au/adt-WU2007.0190.
Annotation:
[Truncated abstract] Hepatocellular carcinoma (HCC) is the major malignancy complicating chronic liver disease. New therapies for the prevention of HCC are required due to the limited success and high tumour recurrence rates of existing treatments. Emerging evidence suggests that HCC arise from the transformation of adult liver progenitor cells (LPCs), which have the capacity to differentiate into hepatocytes and biliary cells during liver regeneration. LPC activation precedes neoplasia in experimental hepatocarcinogenesis. LPCs share antigenic epitopes with HCCs, including α-fetoprotein (AFP) and M2- pyruvate kinase (M2PK). In animal models of hepatocarcinogenesis, attenuation of the LPC response reduces the incidence of HCC following prolonged liver injury via a tumour necrosis factor (TNF) dependent mechanism. As TNF is a pro-inflammatory cytokine, these data suggest that anti-inflammatory agents may be effective in inhibiting LPC activation and hepatocarcinogenesis. Cyclo-oxygenase-2 (COX-2) is an inducible enzyme that mediates the production of many prostaglandins during inflammation and carcinogenesis. Recent investigations show that the administration of selective COX-2 inhibitors (SC2Is) may reduce the incidence of a variety of tumours including breast, colon and skin. The broad aim of this thesis was to conduct a series of detailed studies on the effects of a SC2I on LPC activation and the hepatic pathologies associated with hepatocarcinogenesis in order to test the hypothesis that S2CIs may be a beneficial therapy that can reduce liver injury and pre-neoplastic changes in the choline-deficient, ethionine supplemented (CDE) murine model of hepatocarcinogenesis. Administration of a SC2I (SC-236) significantly inhibited a variety of hepatic cell populations that expand during the first month of the CDE mouse model of hepatocarcinogenesis (a choline deficient, ethionine supplemented diet). Numbers of M2PK-positive LPCs (which are more hepatocytic in morphology and are also COX-2 positive) and inflammatory cells were all significantly reduced by SC-236. In contrast, numbers of A6-positive LPCs (which are more biliary cell-like in morphology and do not express COX-2) were unchanged. ... In summary, these data suggest that COX-2 inhibitors such as SC-236 inhibit LPC activation and a variety of pre-neoplastic liver pathologies as a result of COX-2 dependent and independent mechanisms that may be mediated through inhibition of Akt phosphorylation and induction of apoptosis. Moreover, SC2Is may be useful as preventative treatment strategies for HCC in patients with chronic liver disease.
23
Demasi, Maryanne. „The effects of hypoxia on cyclooxygenase-2 expression and eicosanoid synthesis /“. Title page, table of contents and summary only, 2004. http://web4.library.adelaide.edu.au/theses/09PH/09phd3729.pdf.
Annotation:
Thesis (Ph.D.)--University of Adelaide, Dept. of Medicine and Royal Adelaide Hospital, Rheumatology Unit, 2004.Includes list of publications arising from this thesis. Erratum attached to inside back cover. "25/03/2004." Includes bibliographical references (leaves 185-257).
24
Garlipp, Veridiana Moraes D'Avila Damas. „Efeito agudo do inibidor da fosfodiesterase tipo 5 (sildenafil) na pressão sanguínea arterial durante e após exercício em pacientes submetidos a transplante cardíaco“. Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/5/5131/tde-01022010-165941/.
Annotation:
Introdução: A hipertensão arterial sistêmica (HA) pode estar associada à diminuição na produção e liberação do óxido nítrico derivado do endotélio (NO). O uso do sildenafil leva ao aumento de monofosfato de guanosina cíclica (GMPc), um importante mediador de NO. Contudo, pouco se sabe sobre os efeitos da inibição da fosfodiesterase tipo 5 (PDE5) na monitorização da pressão arterial 24-h (MAPA), pressão arterial durante exercício, noraepinefrina (Nor) e capacidade ao exercício, principalmente após transplante de coração (TX). Métodos: Nós estudamos 22 pacientes pós TX, os quais foram randomizados, tomando dose única de sildenafil (50mg) ou placebo (50mg), aproximadamente uma hora antes de iniciar o protocolo. No dia 1, os pacientes realizaram avaliação clínica, teste cardiopulmonar de caminhada de seis minutos (TES) seguido de teste de esforço cardiopulmonar (TE), Após o término dos testes em esteira, foi colocado o MAPA. Determinamos em repouso (rep), último minuto do TES (6) e pico do TE (Ex): FC (bpm) PAS e PAD (mmHg), VO2(ml/kg/min), Slope VE/VCO2, tempo de exercício (TE, min), distância (TES, Km) e Nor (pg/ml). No dia 2 o protocolo foi repetido, realizando-se o cross-over. Dezessete pacientes apresentavam HA. Resultados: (Pl e Sil respectivamente), Sil reduziu (p<0.05): PAS-rep(138±7 vs 122±18); PAD-rep(83±12 vs 78±12); PAS-6(156± 20 vs 137± 22); PAD-6(82±13 vs 77±14); PAS-Ex(155± 27vs 124±36); PAD-Ex(79±16 vs 66± 16); PAS 24-h(121±10 vs 114±9), PAD 24-h(80±6 vs 76±5), PAS vigília(122±11 vs 115±9), PAD vigília(81± 6 vs 76±5) e PAS noturna(119±12 vs 112±10), PAD noturna(78±7 vs 73±8); e aumentou Nor-repouso(483±165 vs 622±211). Sil não alterou rep, 6 e EX: FC, VO2 e Slope. Conclusão: O ciclo NO-cGMP parece desempenhar papel importante no controle da pressão arterial em TX. Sendo que, a inibição da PDE5 parece apresentar efeitos benéficos no controle da hipertensão arterial em TX, podendo ser utilizada concomitantemente a terapia anti-hipertensiva usual.Background: Systemic hypertension (SH) can be associated with a decrease in endothelium-dependent nitric oxide (NO). Sildenafil determines increment in cyclic guanosine monophosphate (cGMP) that a mediator of NO. However, little is known about the effects of PDE5 inhibition on 24-hour ambulatory (ABP) and exercise blood pressure, noreprinephrine (Nor) and exercise capacity, specially after heart transplantation (HT). Methods: We studied 22 HT pts that on the 1st day underwent a cardiopulmonary (CP) self-controlled treadmill 6walk test(6) and, after, an ECG monitored CP treadmill maximal exercise test(Ex) within 60 and 90 min after oral Sildenafil (Sil,50mg) or placebo(Pl) given at random, and ABP. We determined at basal position(b), last min of 6 and the peak Ex the HR(bpm), SBP and DBP (mmHg), VO2(ml/kg/min), Slope VE/VCO2, exercise time(ET, min), distance(D, Km) and Nor(pg/ml). Also, after CP tests 24-h SBP and DBP were monitored. It was repeated on the 2nd day when the cross-over was done. Seventeen pts had SH. Results: (Pl and Sil respectively), Sil reduced (p<0.05): b- SBP(138±7 vs 122±18); b-DBP(83±12 vs 78±12); 6-SBP(156± 20 vs 137± 22); 6-DBP(82±13 vs 77±14); Ex-SBP(155± 27vs 124±36); Ex-DBP(79±16 vs 66± 16); 24-h SBP(121±10 vs 114±9) and DBP(80±6 vs 76±5), daytime SBP(122±11 vs 115±9) and DBP(81± 6 vs 76±5) and nighttime SBP(119±12 vs 112±10) and DBP(78±7 vs 73±8); and increase b-Nor(483±165 vs 622±211). Sil did not change in b, 6 and EX; HR, Nor, VO2 and Slope. Conclusion: NO-cGMP pathway seems to play a role in blood pressure control in HT. The PDE5 inhibition could have potential beneficial effects on hypertensive HT in addition to antihypertensive therapy.
25
Daniel, Julien. „Etude de l'effet inhibiteur du nicotinamide sur l'activité des lymphocytes B“. Doctoral thesis, Universite Libre de Bruxelles, 2007. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/210719.
Annotation:
Le nicotinamide est un des deux constituants de la vitamine B3. C’est aussi un agent pharmacologique qui a été testé dans le traitement du HIV chez l’homme, et comme traitement contre diverses pathologies. Il présente également des capacités cytoprotectrices dans plusieurs modèles de stress cellulaire et est considéré comme un agent pharmacologique prometteur dans le traitement des maladies de dégénérescence cérébrale d’origine vasculaire. Il module la réponse innée en inhibant notamment la synthèse de molécules pro-inflammatoires. Toutefois, son influence sur la réponse adaptative n’a pas encore été analysée. Le nicotinamide intervient dans la biosynthèse du NAD comme précurseur dans la voie de « sauvetage ». Le rôle du NAD comme coenzyme dans de nombreuses réactions enzymatiques du métabolisme de la cellule est bien connu. De plus, le NAD peut être dégradé par différentes enzymes impliquées dans différentes modifications post-traductionnelles de protéines (PARPs, Sirtuines et MARTs). Le nicotinamide est un produit de la dégradation du NAD mais également un inhibiteur de ces enzymes et constitue donc un outil permettant d’étudier le rôle de ces enzymes dans la transduction de signaux intracellulaires.
Nous avons utilisé le nicotinamide comme un inhibiteur non toxique des différentes enzymes impliquées dans les réactions d’ADP-ribosylation et étudié son effet sur l’activation des lymphocytes B. Le nicotinamide inhibe la prolifération et la différentiation de ces cellules. Il ne module pas les étapes précoces du BCR mais inhibe l’activation des MAPKs et de la kinase Akt. L’inhibition des MAPKs Erk est corrélée avec une réduction de l’expression de la cycline D2 et du marqueur d’activation CD69. L’utilisation d’un inhibiteur des PARPs ne nous a pas permis de reproduire les effets du NAm sur la voie MAPK Erk et CD69. Par contre, le MIBG, un inhibiteur des MARTs inhibe bien la surexpression du CD69 ainsi que la phosphorylation des kinases Erk. Bien que le nicotinamide soit capable d’inhiber l’expression du CD69 in vivo, nos expériences ne nous ont pas permis de moduler la réponse immune adaptative in vivo.
Ceci suggère dès lors que l’utilisation de fortes doses de nicotinamide comme traitement pharmacologique de certaines affections chez l’homme ne devrait pas poser de problème au niveau de la réponse adaptative. De plus, notre mise en évidence des MARTs dans le contrôle de l’activation des lymphocytes B ouvre des perspectives encourageantes pour de nouveaux traitements modulant la réponse adaptative. Cette réponse étant particulièrement impliquée dans certaines maladies auto-immunes, il est potentiellement intéressant de trouver des inhibiteurs de ces enzymes plus puissants que le nicotinamide afin de moduler la réponse immune adaptative in vivo
Doctorat en sciences, Spécialisation biologie moléculaire
info:eu-repo/semantics/nonPublished
26
Ferraz, Daniel Araujo. „Estudo comparativo de fotocoagulação panretiniana com e sem ranibizumabe intravítreo no tratamento da retinopatia diabética proliferativa“. Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/5/5149/tde-21012016-112152/.
Annotation:
Objetivo: Comparar o efeito da terapia da fotocoagulação panretiniana (PFC) associada à injeção intravítrea de Ranibizumabe (RBZ) versus terapia isolada com PFC em pacientes com retinopatia diabética proliferativa (RDP) precoce, virgens de tratamento, com ou sem edema macular diabético (DME) durante 6 meses de acompanhamento. Projeto: Estudo prospectivo intervencionista, randomizado e controlado. Métodos: Sessenta olhos de 30 pacientes com RDP bilateral precoce foram randomizados para o grupo de estudo (GE) que foram tratados com PFC associado a duas injeções de RBZ intravítreo (0.5mg/0.05ml) ou para o grupo controle (GC) tratados apenas com PFC. Mudanças na acuidade visual (AV) corrigida, na sensibilidade ao contraste (SC) e na espessura foveal (EF) foram comparados no início, e nos 1, 3 e 6 meses após o tratamento. Resultados: No GE, a diferença na média da AV do baseline para o mês 6 teve um aumento significativo de + 3,4 letras (p = 0,006) e uma diminuição significativa na EF de - 47.6um (p < 0,001). No GC, a diferença na média da AV teve uma diminuição de - 3,4 letras (p = 0,04) e uma mudança na EF de -3.8 um (p = 0,96). Com relação ao teste de SC dentre os 28 olhos do GE, houve uma melhora no mês 6 em relação ao baseline nos ciclos: 1,5 (p < 0.001) e 3,0 ciclo (p=0.023). Dentre os 30 olhos do GC, não houve uma diferença estatística nos momentos estudados. Conclusão: A injeção intravítrea de RBZ associado com PFC pode ser um tratamento eficaz em olhos de pacientes com RDP precoce e EMDPurpose: To compare the efficacy of therapy with panretinal photocoagulation (PRP) and intravitreal ranibizumab (RBZ) injection versus PRP alone in patients with treatment-naive bilateral non-high risk proliferative diabetic retinopathy (PDR) with and without diabetic macular edema (DME) with a 6-month follow-up. Design: Prospective, interventional, randomized controlled trial. Methods: Sixty eyes of 30 patients with bilateral non-high risk PDR were randomized either to the study group (SG) receiving PRP plus two intravitreal ranibizumab injections (0.5mg/0.05ml), the first one week before and the second four weeks after the PRP or to the control group (CG) receiving PRP alone. Mean change in best-corrected visual acuity (BCVA), contrast sensitivity (CS) and central macular thickness (CMT) were compared at baseline and 1, 3 and 6 months after treatment. Results: Changes from baseline to 6 months showed in the SG an increased in the BCVA by + 3.4 letters (p= 0.006) with a decrease in CMT by - 47.6um (p < 0.001). In the CG, a decrease by - 3.4 letters (p = 0.04) and an decrease by -3.8um (p= 0.96). Regarding the CS in the SG, there was an improvement compared to baseline for the sixth month in the 1.5 (p < 0.001) and 3.0 cycles (p = 0.023). The CG did not show significant results from baseline to month 6. Conclusion: Intravitreal RBZ associated with PRP can be an effective treatment in eyes with non-high risk PDR and DME
27
Preti, Rony Carlos. „Avaliação estrutural e funcional da mácula nos pacientes com retinopatia diabética proliferativa submetidos à panfotocoagulação associada a injeções intravítreas de bevacizumabe“. Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/5/5149/tde-18032013-153010/.
Annotation:
INTRODUÇÃO: O presente estudo avaliou o tratamento com injeções intravítreas de Bevacizumabe (IVB) associadas à panfotocoagulação (PFC) da retina na retinopatia diabética proliferativa (RDP) de alto risco com ou sem edema macular (EM). MÉTODOS: Ensaio clínico randomizado, prospectivo, aberto e mascarado composto por pacientes com Diabetes melitos (DM) tipo 2. A acuidade visual (AV) foi medida com a tabela Early Treatment Diabetic Retinopathy Study e a sensibilidade ao contraste (SC) pela da tabela Vistech Consultants Incorporation 6500. Os pacientes foram submetidos a exame de angiofluoresceinografia para observação de neovascularização retiniana e isquemia macular e à tomografia de coerência óptica (OCT), para se obter a espessura foveal (EF) e o volume macular (VM). Após os exames, um dos olhos do mesmo paciente foi randomizado para realizar somente PFC, grupo controle (GC), e o outro para PFC associado a injeções IVB, grupo de estudo (GE). A hemorragia vítrea (HV) e a presença de complicações também foram avaliadas. RESULTADOS: Dos 42 pacientes incluídos, 35 completaram o estudo. A média de idade foi de 56±8 anos, com predominância do gênero masculino 21 (60%). Vinte e seis (74%) pacientes eram portadores de Hipertensão Arterial Sistêmica com média de duração de 9±10 anos. A média de duração do DM foi de 18±9 anos sendo 23 (66%) usuários de insulina e 21 (68,5%), fácicos. A AV e a SC não demonstraram diferença entre os grupos no total da amostra. O GE demonstrou melhora em comparação ao GC na EF no 1º mês, e no VM nos 1° e 3º meses de seguimento. Quanto aos 12 pacientes com EM bilateral somente a EF demonstrou redução no GE no 1º mês de seguimento. Ao se avaliar os grupos separadamente, o GC apresentou agravamento da AV e SC durante todo seguimento. Houve também aumento da EF nos 1º e 6º meses e VM nos 1º , 3º e 6º meses de seguimento. O GE demonstrou estabilização da AV, SC, EF e VM. Correlacionado às funções visuais, AV com a SC, toda vez que houve piora da AV esta foi acompanhada pelo agravamento da SC em todos os momentos no GC e GE. Quando correlacionadas as AV e SC com as EF e VM, toda vez que a espessura macular aumentava, havia piora da função visual. Dos sete pacientes excluídos do estudo por apresentarem HV, cinco integravam o GC e dois o GE. Não houve aparecimento de catarata, endoftalmite e/ou aumento significativo da pressão ocular. CONCLUSÃO: Na RDP de alto risco, o uso adjuvante de injeções intravítreas de Bevacizumabe associadas à panfotocoagulação da retina pode estabilizar a AV, SC, EF e VM, diminuir a incidência de HV e reduzir a da espessura macular. Em relação à correlação entre as variáveis, quando houve piora da AV, esta foi acompanhada da piora da SC e o aumento da EF e VM causaram piora da AV e SCINTRODUCTION: This study evaluated the treatment with intravitreal injections of Bevacizumab (IVB) associated with panretinal photocoagulation (PRP) in high-risk proliferative diabetic retinopathy (PDR) with or without diabetic macular edema (DME). METHODS: Prospective, open and masked, randomized clinical trial, composed of patients with type 2 Diabetes Mellitus (DM). The visual acuity (VA) was measured with the Early Treatment Diabetic Retinopathy Study charts and the contrast sensitivity (CS) through the chart of Vistech Consultants Incorporation 6500. Patients were submitted to a fluorescein angiography examination to observe retinal neovascularization and macular ischemia and to an optical coherence tomography (OCT) to obtain the foveal thickness (FT) and macular volume (MV). After the tests, one of the eyes from the same patient was randomized to realize only the PRP, the control group (CG), and the other for PRP associated to IVB injections, the study group (SG). Vitreous hemorrhage (VH) and presence of complications were also evaluated. RESULTS: Thirty-five of the forty-two patients included, completed the study. The mean age was 56±8 years, with a predominance of 21 (60%) males. Twenty-six (74%) patients had systemic hypertension with a mean duration of 9±10 years. The mean duration of DM was 18±9 years, of which 23 (66%) were insulin users and 21 (68.5%) were phakic. The VA and CS showed no difference between groups in the total sample. The SG showed improvement compared to the CG in FT for the 1st month, and in MV for the 1st and 3rd month of follow-up. As for the 12 patients with bilateral ME, only the FT showed a reduction in the SG for the 1st month of follow-up. When evaluating the groups separately, the CG showed worsening of VA and CS at all times. There was also an increase of FT for the 1st and 6th months and of MV for the 1st, 3rd and 6th month follow-up. The SG showed stabilization of VA, CS, FT and MV. When correlated to visual functions, VA and CS, a worsening of the VA was accompanied every time by a worsening of the CS in both the CG and SG. When VA and CS are correlated to FT and MV, there was worsening of visual function whenever macular thickness increased. Of the seven patients excluded from the study by presenting VH, 5 belonged to the CG and the 2 to the SG. There was no incidence of cataracts, endophthalmitis and/or significant increase in intraocular pressure. CONCLUSION: In high-risk PDR, intraocular injections of Bevacizumab as an adjuvant treatment to PRP, can stabilize VA, CS, FT and MV, reduce of the incidence of VH and decrease the macular thickness. Regarding the correlation between variables, when there was a worsening of VA, this was accompanied by a worsening of the CS, and an increase in FT and MV caused the worsening of the VA and CS
28
Ávila, Renata. „"Reabilitação neuropsicológica dos processos de memória e das atividades da vida diária em pacientes com doença de Alzheimer leve e moderada"“. Universidade de São Paulo, 2004. http://www.teses.usp.br/teses/disponiveis/5/5160/tde-24082005-150424/.
Annotation:
O efeito da reabilitação neuropsicológica foi verificado em uma amostra de 16 pacientes com diagnóstico de doença de Alzheimer leve e moderado. Após ensaio clínico aberto com rivastigmina por 4 meses, os pacientes foram divididos em três grupos: sessões em grupo, individual e em casa com o cuidador. Os três grupos realizaram o mesmo protocolo de reabilitação, e antes e depois das 22 semanas de tratamento foram avaliados pelos mesmos instrumentos. Os resultados indicaram que as sessões em grupo são mais eficientes para sintomas psiquiátricos; individual para atividades da vida diária; e em casa, dependendo do perfil do paciente e do cuidador, pode ser uma alternativa de tratamentoThe effect of a neuropsychological rehabilitation was tested in a sample of 16 patients with mild and moderate Alzheimer disease. After an open trial with rivastigmine for 4 months, they were divided in 3 groups: group sessions, individualized and at home with a caregiver. All 3 groups fulfilled the same rehabilitation protocol, and just before and after the 22 week period of treatment, all patients were evaluated using the same instruments. The results of the study indicated that group session are more effective for psychiatric symptoms, individualized sessions for activities of daily living training and at home training, depending on the patient's and caregiver's profiles, can be an option for treatment of these patients
29
Eira, Margareth da. „\"Alterações na função renal em pacientes HIV/AIDS tratados com esquemas terapêuticos incluindo indinavir\"“. Universidade de São Paulo, 2004. http://www.teses.usp.br/teses/disponiveis/5/5134/tde-10052005-150439/.
Annotation:
Complicações renais e urológicas incluindo nefrolitíase, cristalúria, cólica renal e lombalgia, são eventos adversos bem conhecidos do indinavir (IDV), um inibidor de protease (IP) largamente utilizado no tratamento de pacientes infectados com o vírus da imunodeficiência humana (HIV). Prévios estudos em ratos demonstraram que o IDV, um potente IP capaz de provocar uma sustentada supressão da carga viral do HIV, induz vasoconstricção renal, diminui a filtração glomerular (RFG) e reduz a excreção urinária de nitrito (NO2-), sugerindo que a vasoconstricção causada pelo IDV deve ser mediada pelo óxido nítrico (NO). Os objetivos deste estudo foram investigar a ocorrência de insuficiência renal (clearance de creatinina < 80ml/min) em pacientes com infecção pelo HIV tratados com terapia anti-retroviral altamente potente incluindo o inibidor de protease IDV, e mensurar a excreção urinária de nitrato (NO3-) nestes pacientes, comparando-os com outro grupo de pacientes tratados com efavirenz (EFV), um inibidor de transcriptase reversa não-análogo de nucleosídeo (NNRTI). No período compreendido entre março de 2000 e outubro de 2003, estudamos 36 pacientes infectados pelo HIV que estavam em terapia com IDV na dose de 800 mg de 8/8 horas por pelo menos 12 meses. Os pacientes foram avaliados para uma variedade de parâmetros clínicos e laboratoriais: idade, peso, tempo de infecção, tempo de uso de IDV, uso de sulfametoxazol-trimetoprim (SMX-TMP) ou sulfadiazina, exames bioquímicos (colesterol total, triglicérides, magnésio, sódio, potássio e creatinina), exame do sedimento urinário, clearance de creatinina, osmolaridade urinária, volume urinário de 24 h, fração de excreção de sódio (FENa), fração de excreção de potássio (FEK) e fração de excreção de água (FEH2O). NO3 urinário foi mensurado em 18 pacientes recebendo terapia anti-retroviral com IDV e 8 pacientes recebendo terapia com EFV. Leucocitúria ocorreu em 78.8% dos pacientes tratados com IDV. Clearance de creatinina diminuído foi observado em 21 pacientes e foi associado com menor peso e uso de derivados de sulfa. Nestes pacientes com diminuição da função renal, também detectamos menor osmolaridade urinária e uma FEH2O mais alta. A excreção urinária de NO3- foi significativamente menor nos pacientes tratados com IDV (908 ± 181) quando comparados aos pacientes do grupo EFV (2247 ± 648, p<0.01). Nossos resultados mostram que insuficiência renal ocorreu em 58% dos pacientes tratados com IDV e foi associada com menor peso corpóreo e uso de derivados de sulfa. A menor excreção urinária de NO3- e as alterações na osmolaridade e FEH2O sugerem que o IDV diminui a produção de óxido nítrico e causa dano tubular, respectivamente. Sugerimos então que os pacientes em uso de IDV sejam monitorados routineiramente para função renal através do clearance de creatinina.Renal and urological complications including nephrolithiasis, crystalluria, renal colic and flank pain are significant side effects of the HIV protease inhibitor indinavir (IDV), and IDV has been widely used in the treatment of human immunodeficiency virus (HIV) infection. Previous studies in rats demonstrated that IDV, a potent protease inhibitor that causes profound and sustained supression of HIV replication, also induces renal vasoconstriction, decreases glomerular filtration rate (GFR) and reduces urinary excretion of nitrite (NO2-), suggesting that IDV-vasoconstriction may be mediated by nitric oxide (NO). The objectives of this study were to investigate the occurrence of renal failure (creatinine clearance <80ml/min) in human HIV patients treated with highy active antiretroviral therapy (HAART), including IDV, and to measure urinary excretion of nitrate (NO3-) in those patients, comparing it with that of another group of patients treated with the non-nucleoside reverse-transcriptase inhibitor efavirenz (EFV). From March 2000 through October 2003, we evaluated 36 patients infected with HIV who was receiving IDV 800 mg q8h for at least 12 months. The patients were assessed for a variety of clinical and laboratory parameters including age, body weight, duration of infection, time of IDV treatment, trimethoprim/sulfamethoxazole (TMP/SMX) or sulfadiazine use, biochemistry (total cholesterol, triglycerides, magnesium, sodium, potassium and creatinine), urinalysis, creatinine clearance, urine osmolality, 24-hour urine volume, fractional excretion of sodium (FENa), potassium (FEK) and water (FEH2O). Urinary NO3 was measured in 18 IDV-treated patients and compared with that of 8 EFV-treated patients. Leukocyturia occurred in 78.8% of the IDV-treated patients. Reduced creatinine clearance was observed in 21 patients and was associated with lower body weight and sulfa-derivated use. In these renal failure patients, we also detected a lower osmolality and a higher FEH2O. Excretion of NO3- was significantly lower in IDV-treated patients (908 ± 181) than in EFV-treated patients (2247 ± 648, p<0.01). Our data show that renal failure occurred in 58% of IDV-treated patients and was associated with lower body weight and sulfa administration. The lower NO3- excretion suggests that this drug decreases nitric oxide production, and the alterations in osmolality and FEH2O indicate that it also causes tubular damage. Based on our findings, we suggest that the renal function of patients under IDV treatment should be closely monitored with creatinine clearance.
30
Dib, Ricardo Anuar. „Avaliação de sintomas e lesões esôfago-gastroduodenais secundários ao uso de antiinflamatórios“. Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/5/5168/tde-08112013-110643/.
Annotation:
Introdução: Os antiinflamatórios não esteróides (AINEs), incluindo a aspirina, são drogas largamente utilizadas para tratamento das doenças inflamatórias e da dor, e que podem causar efeitos colaterais sérios, causando considerável morbidade e mortalidade, relacionadas á doença ulcerosa, duodenal e gástrica, particularmente ao sangramento gastrointestinal. O risco relativo global de complicações gastroduodenais é de três a dez vezes, maior nos usuários de AINEs, quando comparado com indivíduos sadios. Cerca de 25% dos usuários crônicos dos antiinflamatórios não esteroides (AINEs) deverão desenvolver doença ulcerosa, e de 2 a 4% deverão apresentar sangramento ou perfuração. Mais de 17.000.000 de norte americanos utilizam vários tipos de drogas antiinflamatórios não esteróides (AINEs) diariamente e que provocam mais de 100.000 hospitalizações e cerca de 7000 a 10.000 mortes por ano nos Estados Unidos da América do Norte, fazendo desta família de drogas uma das mais comumente usadas em todo planeta. Cerca de 50% das lesões observadas em endoscopias de controle, ocorrem sem que o paciente tenha qualquer tipo de sintoma. Acredita-se que houve recrudescimento da prevalência de lesões digestivas pela substituição dos antiinflamatórios COX-2 pelos antiinflamatórios tradicionais, principalmente pela ausência de cuidados na prevenção deste tipo de ocorrência, em populações consideradas de risco. Objetivos: a) avaliar a prevalência de lesões e complicações digestivas secundárias ao uso de AINEs; b) qual é o perfil clínico deste paciente atendido em razão de queixas digestivas e a relação destas com os achados endoscópicos. Materiais e métodos: estudo aberto, prospectivo, multicêntrico avaliando consecutivamente 1.231 pacientes submetidos a exame de endoscopia digestiva alta em virtude de queixas digestivas, única ou associadas, como: 1) pirose; 2) dor epigástrica; 3) dor abdominal; 4) náusea; 5) vômito. Antes da realização do exame de endoscopia digestiva alta, os pacientes respondiam a questionário cujo objetivo era avaliar o início e o tipo de queixa clínica, o uso de medicamentos e possíveis complicações associadas como sangramento digestivo. Os critérios de inclusão foram: pacientes de ambos os sexos com idade mínima de 18 anos e que tivessem sintomas prévios iniciados, no máximo, há 14 dias antes da realização do exame de endoscopia digestiva alta. Os critérios de exclusão foram os de pacientes que se recusaram a participar do estudo e/ou de assinar o Termo de Consentimento Livre e Esclarecido, os incapazes de responder ao questionário, os com idade inferior aos 18 anos, os pacientes que já haviam realizado cirurgia gástrica e pacientes portadores de insuficiência renal ou hepática. Resultados: Foram avaliados 1.213 pacientes de 18 a 82 anos sendo que 65% destes eram do sexo feminino, 13,1% eram fumantes e 15,6% referiam ingestão de bebidas alcoólicas. A utilização de AINEs foi mais frequente no sexo feminino, porém número de complicações foi maior nos pacientes do sexo masculino (sangramentos foi duas vezes maior; p=0,045 e a ocorrência de úlcera quase 1,5 vezes maior; p=0,041). Os principais sinais e sintomas relatados foram epigastralgia e pirose (67% e 62%, respectivamente). Os 1.213 pacientes foram alocados em dois grupos: Grupo I - AINE composto por 228 (18,8%) e o Grupo II - Não AINEs (NAINEs) por 985 (81,2%) pacientes.. O exame de endoscopia digestiva alta foi normal em 3,9% dos pacientes do grupo I e em 10,7% dos do grupo II (p< 0,001). A probabilidade de um paciente que não utiliza AINE ter endoscopia digestiva alta normal é 2,5 vezes maior quando comparado aos que utilizaram AINEs (p=0,001). As presenças de lesões erosivas ou ulceradas no estômago e duodeno também foram mais frequentes nos pacientes do Grupo I quando comparado aos do Grupo II. Observa-se que é maior a incidência de lesões, tanto erosivas quanto ulceradas no estômago quando comparadas ao duodeno (erosões: 49,12% vs 13,60 respectivamente, p=0,001; úlceras: 14,04% vs 11,84% respectivamente, p= 0,05). O risco de hemorragia digestiva, 12 vezes maior (6,14% vs 0,51%) nos pacientes que fizeram uso de AINEs sendo o estômago o sítio de maior prevalência de sangramento. Não se observou diferença estatística quando analisada a presença de esofagite erosiva nos dois grupos. Conclusões: Evidenciamos frequência maior de úlcera gástrica, úlcera duodenal e sangramento digestivo nos pacientes que utilizaram AINEs. Não foram encontradas relações entre os achados endoscópicos e os sintomas dispépticos. Não observamos influência dos AINEs no aparecimento de esofagite erosivaIntroduction: The non steroidal anti-inflammatory drugs (NSAID), including aspirin, are drugs widely used in the treatment of inflammatory diseases and pain. This use may cause serious side-effects, leading to considerable morbidity and mortality related to ulcer, duodenal and gastric disease, especially gastrointestinal bleeding. The overall relative risk of gastroduodenal complications is three to ten times higher in users of NSAID, compared to healthy individuals. Around 25% of the chronic users of non steroidal anti-inflammatory drugs (NSAID) will develop ulcer disease, and 2 to 4% will present bleeding or perforation. More than 17,000,000 North Americans use several kinds of non steroidal anti-inflammatory drugs (NSAID) on a daily basis. This causes more than 100,000 hospitalizations and from 7,000 to 10,000 deaths every year in the USA, which makes this drug one of the most commonly used on the planet. About 50% of the lesions observed in endoscopies occur without any kind of symptom. It is believed that there was an increase in the prevalence of digestive lesions due to the replacement of COX-2 anti-inflammatory drugs with traditional anti-inflammatory drugs, especially because of the lack of preventive care of this kind of occurrence in at-risk populations. Goals: a) Evaluate the prevalence of lesions and digestive complications, secondary to the use of NSAID; b) Evaluate the clinical profile of the patient seen for digestive complaints and the relation of these complaints with the endoscopic findings. Materials and Methods: Prospective, multi-centric, open study, evaluating consecutively 1,231 patients who underwent upper gastrointestinal endoscopy exam due to digestive complaints in isolation or associated, such as: 1) pyrosis; 2) epigastric pain; 3) abdominal pain; 4) nausea; 5) vomiting. Before performing the exam of upper gastrointestinal endoscopy, patients answered a questionnaire whose goal was to evaluate the onset and kind of clinical complaint, the use of medication and possible complications associated to digestive bleeding. The inclusion criteria were: Patients of both sexes with the minimum age of 18 and whose symptoms had begun up to 14 days before undergoing the upper gastrointestinal endoscopy. Exclusion criteria: patients who refused to participate in the study and/ or who refused to sign the Informed Consent Term, the ones who were unable to respond to the questionnaire, the ones who were under 18 years old, patients who had undergone a previous gastric surgery and patients with kidney or hepatic failure. Results: 1,213 patients with ages ranging from 18-82 were evaluated, 65% of which were female and 13,1% were smokers, 15,6% mentioned they ingested alcoholic beverages. The use of NSAID was more frequent among females. However, the number of complications was higher among males (bleeding occurred twice as much; p=0,045 and the occurrence of ulcer was almost 1,5 times higher; p=0,041). The main signs and symptoms reported were epigastralgia and pyrosis (67% and 62%). The 1,213 patients were divided into two groups: Group I- NSAID, made up by 228 (18,8%) and Group II- Non NSAID, made up by 985 patients (81,2%). The upper gastrointestinal endoscopy was normal in 3,9% of the patients in Group I and in 10,7% of the patients in Group II (p<0,001). A patient who does not use NSAID will be 2,5 times more likely to have normal upper gastrointestinal endoscopy than the one who used NSAID (p=0,001). The presence of erosive or ulcer lesions in the stomach and duodenum was more frequent in Group I patients when compared to those of Group II. It is observed that the incidence of lesions in the stomach, both erosive and ulcer is higher when compared to the duodenum (erosions: 49,12% vs. 13,60, p=0,001; ulcers: 14,04% vs. 11,84, p= 0,05). The risk of digestive bleeding is 12 times higher (6,14% vs. 0,51%) in patients who used NSAID, and the stomach is the site with higher prevalence of bleeding. No statistic difference was observed when the presence of erosive esophagitis in both groups was analyzed. Conclusions: We observed that the frequency of gastric ulcer, duodenal ulcer and digestive bleeding was higher in patients who used NSAID. Relations between the endoscopic findings and the dyspeptic symptoms were not found. The influence of NSAIDs on the appearance of erosive esophagitis was not observed
31
Roach, Denise Margaret. „Upregulation of matrix metalloproteinases -2 and -9 and type IV collagen degradation in skeletal muscle reperfusion injury“. Thesis, 2002. http://hdl.handle.net/2440/38409.
Annotation:
Includes bibliographical references (leaves 292-352)xvi, 352 leaves
Determines the role of matrix metalloproteinases, MMP-2 and MMP-9 in reperfusion injury following skeletal muscle ischaemia; and, whether inhibition of MMPs by doxycycline protects against tissue damage.
Thesis (M.D.) -- University of Adelaide, Dept. of Surgery, 2002
32
„Study of neuroprotective effect of cryptotanshinone, an acetylcholinesterase inhibitor, in cell and animal models“. Thesis, 2009. http://library.cuhk.edu.hk/record=b6074975.
Annotation:
Alzhemier's disease (AD) is a common form of dementia which is characterized by the deposition of amyloids in affected neurons and a cholinergic neurotransmission deficit in the brain. Current therapeutic intervention for AD is primarily based on inhibition of brain acetylcholinesterase (AChE) to restore the brain acetylcholine level. Cryptotanshinone (CT) is a diterprene which is extracted from the root of Salvia miltiorrhiza, an herb that is commonly prescribed in Chinese medicine to treat cardiovascular disease. The present study is aimed at verifying CT's property as an AChE inhibitor using different models. By AChE activity assay, CT was found to be a dual inhibitor which inhibits both human acetylcholinesterase (AChE) and butylcholinesterase (BuChE) with similar IC50. CT inhibited human AChE in a reversible manner, and the inhibition showed the characteristics of mixed-type. To human BuChE, CT is an uncompetitive inhibitor. CT can also inhibit AChE from rat cortical neurons. Apart from AChE inhibition, CT was demonstrated to have ameliorating effect on glutamate excitotoxicity, which is a cause of neuron death in AD. Further study showing that CT treatment can reduce cellular tau phosphorylation, which is the downstream effector of glutamate-induced excitotoxicity. In animal model, the effect of CT on learning impairment in scopolamine-treated rats was also evaluated by the acquisition protocol of Morris water maze. The task learning ability of scopolamine-treated rats was significantly reversed by CT, and the CT-fed rats were able to develop spatial searching strategy comparable to the control animals. Chronic administration of CT at effective doses did not cause significant hepatotoxicity. Cholinergic side effect of muscle weakness was not observed in CT treated rats. On the contrary CT was found to increase the locomotor activity of NIH mice in forced swimming test through reducing the lactic acid in the circulation. Data in this study gives further support on CT's potential as a therapeutic drug for treating AD.by Wong, Kin Kwan Kelvin.
Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2009.
Includes bibliographical references (leaves 144-167).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
33
„CNS target delivery of acetylcholine esterase inhibitors via intranasal administration: pilot studies with tacrine and Its dimers“. 2013. http://library.cuhk.edu.hk/record=b5884375.
Annotation:
Qian, Shuai.Thesis (Ph.D.)--Chinese University of Hong Kong, 2013.
Includes bibliographical references (leaves 265-294).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
34
„Effect of phytochemicals on estrogen biosynthesis in human breast cancer and placental cells“. Thesis, 2005. http://library.cuhk.edu.hk/record=b6074044.
Annotation:
A breast cancer cell line stably transfected with the CYP19 gene had been employed for aromatase inhibition. Among the phytochemicals tested, the major dietary flavonoids, such as genistein and daidzein, produced very weak inhibition. On the other hand, the red clover isoflavone biochanin A, the hydroxychalcone butein and the red grape phytoalexin resveratrol were found to be effective aromatase inhibitors. Cell proliferation assay had shown that they could inhibit ER-positive cell proliferation induced by testosterone, and the inhibitory effect was specifically attributed to the reduction of estrogen synthesis. In another breast cancer cell line SK-BR-3, resveratrol, biochanin A and genistein inhibited CYP19 both in enzyme and promoter I.3/II transcriptional levels. The element responsible for the inhibition of aromatase by these phytoestrogens should fall within the region between -556 to -446 by upstream of exon II.Breast cancer is one of the most common cancers affecting women. Estrogen plays an important role in breast cancer initiation and development. The majority of breast tumors are initially dependent upon estrogen to support their growth. Most breast cancers occur in the postmenopausal period. However, the intra-tumoral estradiol (E2) is maintained at a high level equivalent to the pre-menopausal status. High intra-tumoral E2 level in postmenopausal women is sustained by the biosynthesis of estrogens in the tumorous tissue.
Genistein and Biochanin A, ranged from 0.1 to 10 muM, might act as estrogen agonist and induced aromatase activity and promoter I.1 transactivation in ERalpha-transfected SK-BR-3 cells. (Abstract shortened by UMI.)
The aromatase enzyme, CYP19, belongs to a family of P450 enzyme. As a final rate-limiting step in estrogen biosynthesis, it catalyzes the conversion of C 19 steroids to estrogens. The expression of CYP19 is tissue-specific, and is regulated by alternate promoter usage. The use of aromatase inhibitors for breast cancer treatment has become a major therapeutic approach.
The consumption of some phytochemicals protects against breast cancer. Yet the mechanisms are far from clear. In my present study, various phytochemicals, including phytoestrogens, monoterpenes and carotenoids, were evaluated for their effect on aromatase.
Wang Yun.
"July 2005."
Adviser: Lai-Kwok Leung.
Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3716.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (p. 145-169).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract in English and Chinese.
School code: 1307.
35
„Antifungal activities of metergoline, purpurin and baicalein on Candida species“. Thesis, 2010. http://library.cuhk.edu.hk/record=b6075072.
Annotation:
Baicalein is known to be a potent antifungal agent and induces programmed cell death in Candida albicans. In the present study, we found that baicalein also inhibited the growth of C. krusei isolates. The minimal inhibitory concentrations of baicalein against eight C. krusei isolates were 1.35--2.70 microg/ml. One-hour exposure to baicalein elicited a consistent and moderate post-antifungal effect on the C. krusei isolates. Further flow cytometric study demonstrated a depolarization of mitochondrial membrane potential. However, both the levels of reactive oxygen species and DNA fragmentation were not significantly changed after baicalein treatment in C. krusei. It can be concluded that the antifungal activity of baicalein was mitochondria-dependent in both C. krusei and C. albicans, but the antifungal mechanism was different. Reactive oxygen species may not play a direct role and baicalein does not initiate programmed cell death or apoptosis in C. krusei. The structure-activity relationship study showed that the three hydroxyl groups in baicalein were essential for its antifungal potency.Candidiasis has become a serious infection with very high mortality and morbidity in the world if not providing effective treatments. However, due to clinical limitation and resistance of the current antifungal agents, there is an urgent need to search for novel antifungals. In this study, after screening a compound library (n=400) for antifungal activity, three members (metergoline, purpurin and baicalein) were chosen for further study. Their antifungal characteristics and the antifungal mechanisms were investigated.
Metergoline, a serotonin receptor antagonist, was found to have potent antifungal activity against the intrinsically fluconazole-resistant human fungal pathogen Candida krusei. The minimal inhibitory concentration and minimal fungicidal concentration of metergoline against C. krusei were 4 microg/ml and 8 microg/ml respectively. Metergoline induced post-antifungal effect. Significant synergism was found in combination of metergoline with amphotericin B by a checkerboard assay, which may be due to the perturbation of cell permeability and increase in the intracellular accumulation of antifungal agents. Metergoline also inhibited extracellular phospholipase production in C. krusei. To gain insights into the mechanisms, intracellular changes that accompany apoptosis were examined by flow cytometry and spectrophotometry. The results showed an increase in the level of reactive oxygen species, depolarization of mitochondrial membrane potential, phosphatidylserine externalization, and positive terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labelling in the metergoline-treated C. krusei . Taken together, we conclude that metergoline may promote apoptosis in C. krusei through reactive oxygen species production and perturbation in mitochondrial homeostasis, implying its antifungal potential to treat candidiasis.
The antifungal activity of purpurin, a natural red anthraquinone pigment in madder root (Rubia tinctorum L.), was evaluated against Candida isolates by a broth microdilution assay. The minimal inhibitory concentrations of purpurin against Candida species isolates were 1.28--5.12 microg/ml. Mechanistic studies indicated that purpurin inhibited energy-dependent efflux pumps of Candida isolates. Furthermore, purpurin demonstrated a depolarization of mitochondrial membrane potential, suggesting a possible linkage of the antifungal mechanism of purpurin to Candida apoptosis.
Kang, Kai.
Adviser: Fong Wing Ping.
Source: Dissertation Abstracts International, Volume: 73-02, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 98-123).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
36
Zhang, Shuo. „Mechanisms of proliferation inhibition and apoptosis induced by vitamin E compounds and cyclooxygenase inhibitors in human breast cancer cells“. Thesis, 2004. http://hdl.handle.net/2152/2107.
37
„The effects of phosphodiesterase inhibitors on rat mast cells“. 2005. http://library.cuhk.edu.hk/record=b5892476.
Annotation:
Kam Man Fai Afia.Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (leaves [195]-224).
Abstracts in English and Chinese.
Abstract --- p.i
Acknowledgement --- p.v
Publications --- p.vi
Abbreviations --- p.vii
Chapter 1. --- Introduction --- p.1
Chapter 1.1 --- The Mast Cell --- p.2
Chapter 1.1.1 --- Historical Perspective --- p.2
Chapter 1.1.2 --- Mast Cell Origin and Development --- p.3
Chapter 1.1.3 --- Mast Cell Heterogeneity --- p.5
Chapter 1.1.3.1 --- Rodent Mast Cell Heterogeneity --- p.5
Chapter 1.1.3.2 --- Human Mast Cell Heterogeneity --- p.7
Chapter 1.1.4 --- Mast Cell Mediators --- p.10
Chapter 1.1.4.1 --- Preformed Mediators --- p.11
Chapter 1.1.4.2 --- Newly Synthesized Lipid Mediators --- p.14
Chapter 1.1.4.3 --- Cytokines --- p.16
Chapter 1.1.5 --- Mast Cell Activation --- p.17
Chapter 1.1.5.1 --- Immunological Activation --- p.19
Chapter 1.1.5.1.1 --- FcεIR Activation and Protein Tyrosine Phosphorylation --- p.19
Chapter 1.1.5.1.2 --- Activation of Phospholipases --- p.20
Chapter 1.1.5.1.3 --- The Role of Calcium --- p.22
Chapter 1.1.5.1.3.1 --- Intracellular Calcium Mobilization --- p.23
Chapter 1.1.5.1.3.2 --- Calcium Influx --- p.24
Chapter 1.1.5.1.3.3 --- Mechanisms of Action of Calcium in Mast Cells --- p.28
Chapter 1.1.5.1.4 --- The Role of G-proteins --- p.30
Chapter 1.1.5.1.5. --- The Role of Cylic AMP --- p.33
Chapter 1.1.5.1.2.1 --- Mechanisms of Action of Cyclic AMP in Mast Cells --- p.36
Chapter 1.1.5.1.2.2 --- Implications for the Inhibitory Role of Cyclic AMP in Mast Cell Activation --- p.37
Chapter 1.2 --- The Cyclic Nucleotide Phosphodiesterases --- p.39
Chapter 1.2.1 --- Introduction --- p.39
Chapter 1.2.2 --- Classification and Structure --- p.41
Chapter 1.2.3 --- Distribution and Physiological Functions of the Different PDE Families --- p.45
Chapter 1.2.4 --- Phosphodiesterase Inhibitors --- p.49
Chapter 1.2.4.1 --- Non-selective PDE Inhibitors --- p.50
Chapter 1.2.4.2 --- Selective PDE Inhibitors --- p.52
Chapter 1.2.4.2.1 --- PDE1 and PDE2 Inhibitors --- p.52
Chapter 1.2.4.2.2 --- PDE3 Inhibitors --- p.53
Chapter 1.2.4.2.3 --- PDE4 Inhibitors --- p.54
Chapter 1.2.4.2.4.1 --- PDE5 Inhibitors --- p.56
Chapter 2. --- Materials and Methods --- p.59
Chapter 2.1 --- Materials --- p.60
Chapter 2.1.1 --- Drugs --- p.60
Chapter 2.1.1.1 --- Phosphodiesterase Inhibitors --- p.60
Chapter 2.1.1.2 --- Mast Cell Secretagogues --- p.61
Chapter 2.1.2 --- Materials for Rat Peritoneal Mast Cell Experiments --- p.61
Chapter 2.1.2.1 --- Materials for Rat Sensitization --- p.61
Chapter 2.1.2.2 --- Materials for Buffers --- p.62
Chapter 2.1.2.3 --- Materials for Histamine Assay --- p.62
Chapter 2.1.2.4 --- Miscellaneous --- p.63
Chapter 2.1.3 --- Materials for RBL-2H3 Cell Line Experiments --- p.63
Chapter 2.1.3.1 --- Materials for Cell Culture --- p.63
Chapter 2.1.3.2 --- Materials for Cell Sensitization and Enzyme Release --- p.64
Chapter 2.1.3.3 --- Materials for β-Hexosaminidase Assay --- p.64
Chapter 2.1.3.4 --- Miscellaneous --- p.64
Chapter 2.2 --- Rat Peritoneal Mast Cell Experiments --- p.65
Chapter 2.2.1 --- Preparation of Buffers --- p.65
Chapter 2.2.2 --- Preparation of Stock Solutions --- p.66
Chapter 2.2.2.1 --- Mast Cell Secretagogue Stock Solutions --- p.66
Chapter 2.2.2.2 --- Phosphodiesterase Inhibitor Stock Solutions --- p.66
Chapter 2.2.3 --- Animals and Cell Isolation --- p.71
Chapter 2.2.3.1 --- Animals --- p.71
Chapter 2.2.3.2 --- Sensitization of Animals --- p.71
Chapter 2.2.3.3 --- Cell Isolation --- p.71
Chapter 2.2.3.4 --- Cell Purification --- p.72
Chapter 2.2.3.5 --- Determination of Cell Number and Viability --- p.73
Chapter 2.2.4 --- General Protocol for Histamine Release and Histamine Measurement --- p.75
Chapter 2.2.4.1 --- Histamine Release --- p.75
Chapter 2.2.4.2 --- Spectrofluorometric Determination of Histamine Content --- p.76
Chapter 2.2.4.2.1 --- Manual Histamine Assay --- p.76
Chapter 2.2.4.2.2 --- Automated Histamine Assay --- p.78
Chapter 2.2.4.3 --- Calculation of Histamine Levels --- p.78
Chapter 2.2.4.4 --- Presentation and Statistics --- p.79
Chapter 2.3 --- RBL-2H3 Cell Line Experiments --- p.80
Chapter 2.3.1 --- Preparation of Stock Solutions --- p.80
Chapter 2.3.2 --- Preparation of Materials for Enzyme Release and Assay --- p.81
Chapter 2.3.2.1 --- Cell Culture --- p.81
Chapter 2.3.2.2 --- Preparation of Cells for β-Hexosaminidase Release Experiments --- p.82
Chapter 2.3.2.3 --- β-Hexosaminidase Release --- p.82
Chapter 2.3.2.4 --- β-Hexosaminidase Assay --- p.83
Chapter 3. --- Effects of Phosphodiesterase Inhibitors on Mediator Release from Rat Mast Cells --- p.84
Chapter 3.1 --- Introduction --- p.85
Chapter 3.2 --- Materials and Methods --- p.87
Chapter 3.2.1 --- Rat Peritoneal Mast Cells --- p.87
Chapter 3.2.1.1 --- Experiments Employing Immunological Stimulus in RPMCs --- p.87
Chapter 3.2.1.2 --- Experiments Employing Non-Immunological Stimuli in RPMCs --- p.88
Chapter 3.2.2 --- Rat Basophilic Leukemia Cells --- p.88
Chapter 3.3 --- Results --- p.89
Chapter 3.3.1 --- Rat Peritoneal Mast Cells --- p.89
Chapter 3.3.1.1 --- Immunologically Activated Rat Peritoneal Mast Cells --- p.89
Chapter 3.3.1.1.1 --- Effects of Non-Selective PDE Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.89
Chapter 3.3.1.1.2 --- Effects of Selective PDE1 and PDE2 Inhibitors on Anti-IgE- Mediated Histamine Release from RPMCs --- p.90
Chapter 3.3.1.1.3 --- Effects of Selective PDE3 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.90
Chapter 3.3.1.1.4 --- Effects of Selective PDE4 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.91
Chapter 3.3.1.1.5 --- Effects of Selective PDE5 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.91
Chapter 3.3.1.2 --- Non-Immunologically Activated Rat Peritoneal Mast Cells --- p.92
Chapter 3.3.1.2.1 --- Effects of Selective PDE Inhibitors on Compound 48/80- Mediated Histamine Release from RPMCs --- p.92
Chapter 3.3.1.2.2 --- Effects of Selective PDE Inhibitors on Histamine Release from RPMCs Stimulated by Calcium Ionophores --- p.93
Chapter 3.3.2 --- Rat Basophilic Leukemia Cells --- p.93
Chapter 3.3.2.1 --- Effects of Non-Selective PDE Inhibitors on Antigen-Mediated β-Hexosaminidase Release from RBL-2H3 Cells --- p.93
Chapter 3.3.2.2 --- Effects of Selective PDE Inhibitors on Antigen-Mediated β-Hexosaminidase Release from RBL-2H3 Cells --- p.94
Chapter 3.4 --- Discussion --- p.95
Chapter 3.4.1 --- Rat Peritoneal Mast Cells --- p.95
Chapter 3.4.1.1 --- Immunologically Activated RPMCs --- p.95
Chapter 3.4.1.2 --- Non-Immunologically Activated RPMCs --- p.99
Chapter 3.4.2 --- Rat Basophilic Leukemia Cells --- p.103
Chapter 4. --- Combined Effects of Selective Phosphodiesterase Inhibitors on Immunologically Induced Histamine from Rat Mast Cells --- p.143
Chapter 4.1 --- Introduction --- p.144
Chapter 4.2 --- Materials and Methods --- p.144
Chapter 4.2.1 --- Simultaneous Addition of PDE3 and PDE4 Inhibitors --- p.145
Chapter 4.2.2 --- Sequential Addition of PDE3 and PDE4 Inhibitors --- p.145
Chapter 4.3 --- Results --- p.146
Chapter 4.3.1 --- Effects of the Selective Inhibitors for PDE3 and PDE4 Alone: Calculation of the Expected Inhibition Curve --- p.146
Chapter 4.3.2 --- Effects of the Simultaneous Addition of PDE3 and PDE4 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.148
Chapter 4.3.2.1 --- Rolipram and Siguazodan --- p.148
Chapter 4.3.2.2 --- Ro 20-1724 and Siguazodan --- p.149
Chapter 4.3.2.3 --- Rolipram and Quazinone --- p.149
Chapter 4.3.2.4 --- Ro 20-1724 and Quazinone --- p.150
Chapter 4.3.3 --- Effects of the Sequential Addition of PDE3 and PDE4 Inhibitors on Anti-IgE-Mediated Histamine Release from RPMCs --- p.150
Chapter 4.3.3.1 --- Rolipram and Siguazodan --- p.150
Chapter 4.3.3.2 --- Ro 20-1724 and Siguazodan --- p.151
Chapter 4.3.3.3 --- Rolipram and Quazinone --- p.151
Chapter 4.3.3.4 --- Ro 20-1724 and Quazinone --- p.152
Chapter 4.4 --- Discussion --- p.153
Chapter 5. --- Future Directions --- p.191
Chapter 5.1 --- Future Directions --- p.192
References --- p.195
38
„Mechanisms underlying chemopreventive effect of celecoxib in gastric carcinogenesis“. 2006. http://library.cuhk.edu.hk/record=b5893014.
Annotation:
Chu Wai Kit.Thesis (M.Phil.)--Chinese University of Hong Kong, 2006.
Includes bibliographical references (leaves 87-96).
Abstracts in English and Chinese.
Acknowledgments --- p.ii
Publication --- p.iii
List of Abbreviations --- p.iv
List of Tables --- p.v
List of Figures --- p.vi
Abstract --- p.vii
摘要 --- p.x
Table of Contents --- p.xii
Chapter Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Epidemiology of gastric cancer --- p.1
Chapter 1.2 --- Risk factors associated with gastric cancer --- p.7
Chapter 1.3 --- Prevention of Gastric Cancer --- p.9
Chapter 1.4 --- H. pylori eradication and gastric cancer development --- p.11
Chapter 1.5 --- Non-steroidal anti-inflammatory drugs and gastric cancer prevention --- p.13
Chapter 1.6 --- COX-2 independent pathway --- p.14
Chapter 1.7 --- Animal model of gastric cancer --- p.15
Chapter 1.8 --- Microarray system --- p.16
Chapter 1.9 --- Hypothesis --- p.18
Chapter 1.10 --- Aim of study --- p.19
Chapter Chapter 2 --- Chemoprevention of gastric cancer by celecoxib --- p.20
Chapter 2.1 --- Introduction --- p.20
Chapter 2.2 --- Material and Methods --- p.22
Chapter 2.2.1 --- Animals --- p.22
Chapter 2.2.2 --- Chemicals --- p.22
Chapter 2.2.3 --- Study design --- p.23
Chapter 2.2.4 --- Cell Culture --- p.24
Chapter 2.2.5 --- Celecoxib treatment --- p.24
Chapter 2.2.6 --- Cell proliferation assay --- p.25
Chapter 2.3 --- Results --- p.26
Chapter 2.3.1 --- Chemoprevention of gastric cancer by celecoxib in rats --- p.26
Chapter 2.3.2 --- Effects of celecoxib on growth of human gastric cancer cells --- p.29
Chapter 2.4 --- Discussion --- p.30
Chapter 2.4.1 --- MNNG induced gastric cancer effectively --- p.30
Chapter 2.4.2 --- Celecoxib significantly suppressed gastric carcinogenesis in rats --- p.31
Chapter 2.4.3 --- Celecoxib inhibited the growth of MKN 45 in a concentration-dependent manner --- p.31
Chapter 2.4.4 --- Celecoxib may exert its anti-tumor property through COX independent pathway --- p.32
Chapter Chapter 3 --- Gene expression profiles of celecoxib treated rat gastric tumor and human gastric cells --- p.34
Chapter 3.1 --- Introduction --- p.34
Chapter 3.2 --- Material and Methods --- p.34
Chapter 3.2.1 --- RNA extraction --- p.34
Chapter 3.2.2 --- Target preparation and Array hybridization --- p.35
Chapter 3.2.3 --- Post-hybridization processing and Scanning --- p.36
Chapter 3.2.4 --- Microarray data analysis --- p.36
Chapter 3.2.5 --- Quantitative RT-PCR --- p.37
Chapter 3.3 --- Results --- p.39
Chapter 3.3.1 --- Gene expression profiles of rat gastric tumors --- p.39
Chapter 3.3.1.1 --- Genes differentially expressed in MNNG induced gastric tumors --- p.39
Chapter 3.3.1.2 --- Genes differentially expressed in celecoxib treated group --- p.42
Chapter 3.3.1.3 --- Mechanisms underlying chemoprevention of celecoxib --- p.43
Chapter 3.3.2 --- Verification of gene expression by quantitative RT-PCR --- p.55
Chapter 3.3.3 --- Confirmation of the gene expression profiles in human by quantitative RT-PCR --- p.59
Chapter 3.4 --- Discussions --- p.63
Chapter Chapter 4 --- Effects of celecoxib on Akt pathway in gastric cancer cells --- p.68
Chapter 4.1 --- Introduction --- p.68
Chapter 4.2 --- Material and methods --- p.72
Chapter 4.2.1 --- Protein extraction --- p.72
Chapter 4.2.2 --- Western blotting --- p.72
Chapter 4.3 --- Results --- p.74
Chapter 4.3.1 --- Expression of the Akt pathway after treatment with celecoxib --- p.74
Chapter 4.4 --- Discussions --- p.78
Chapter Chapter 5 --- Conclusion --- p.82
References --- p.87
39
„In vitro antioxidant and anti-angiogenic effects of mushroom water extracts“. 2011. http://library.cuhk.edu.hk/record=b5894512.
Annotation:
Lai, Tsz Ching.Thesis (M.Phil.)--Chinese University of Hong Kong, 2011.
Includes bibliographical references (leaves 121-136).
Abstracts in English and Chinese.
Acknowledgements
Abstract
摘要
Content
List of tables
List of figures
List of abbreviations
Chapter Chapter 1: --- Introduction --- p.1
Chapter 1.1 --- Introduction of food market trends in Hong Kong and mushroom productivity in the world --- p.1
Chapter 1.1.1 --- Agrocybe aegerita --- p.1
Chapter 1.1.2 --- Pleurotus spp --- p.2
Chapter 1.1.3 --- Pholiota nameko --- p.3
Chapter 1.2 --- Objectives --- p.5
Chapter Chapter 2: --- Chemical assays for in vitro antioxidative properties of mushroom extracts --- p.6
Chapter 2.1 --- Introduction --- p.6
Chapter 2.1.1 --- Reactive oxygen species (ROS) --- p.6
Chapter 2.1.1.1 --- Definition of ROS --- p.6
Chapter 2.1.1.2 --- Sources of ROS --- p.6
Chapter 2.1.1.2.1 --- Endogenous sources of ROS --- p.6
Chapter 2.1.1.2.2 --- Exogenous sources of ROS --- p.8
Chapter 2.1.1.3 --- Damaging effects of ROS --- p.8
Chapter 2.1.2 --- Antioxidants --- p.10
Chapter 2.1.2.1 --- Mechanism of action --- p.10
Chapter 2.1.2.2 --- Sources of antioxidants --- p.11
Chapter 2.1.2.2.1 --- Dietary antioxidants --- p.11
Chapter 2.1.2.2.2 --- Antioxidants in edible mushrooms --- p.12
Chapter 2.1.2.2.3 --- Phenolic compounds in mushrooms --- p.13
Chapter 2.2 --- Materials and Methods --- p.16
Chapter 2.2.1 --- Materials --- p.16
Chapter 2.2.1.1 --- Mushroom fruiting bodies --- p.16
Chapter 2.2.2 --- Principles of Methods and Experimental Protocols --- p.17
Chapter 2.2.2.1 --- Sample preparation --- p.17
Chapter 2.2.2.2 --- Evaluation of antioxidant capacity --- p.18
Chapter 2.2.2.2.1 --- DPPH radical scavenging activity --- p.18
Chapter 2.2.2.2.2 --- Superoxide anion scavenging activity --- p.19
Chapter 2.2.2.2.3 --- Hydroxyl radical scavenging activity --- p.20
Chapter 2.2.2.2.4 --- Hydrogen peroxide scavenging activity --- p.22
Chapter 2.2.2.3 --- Determination of phenolic compounds --- p.24
Chapter 2.2.2.3.1 --- Total phenolic content --- p.24
Chapter 2.2.2.3.2 --- Identification of phenolic acids --- p.25
Chapter 2.2.3 --- Statistical analysis --- p.27
Chapter 2.3 --- Results and Discussion --- p.28
Chapter 2.3.1 --- Extraction yield --- p.28
Chapter 2.3.2 --- Evaluation of antioxidant capacity --- p.29
Chapter 2.3.2.1 --- DPPH radical scavenging activity --- p.29
Chapter 2.3.2.2 --- Superoxide anion scavenging activity --- p.31
Chapter 2.3.2.3 --- Hydroxyl radical scavenging activity --- p.33
Chapter 2.3.2.4 --- Hydrogen peroxide scavenging activity --- p.35
Chapter 2.3.2.5 --- Comparison of the effective concentrations (EC50) of mushroom water extracts in different antioxidant assays --- p.37
Chapter 2.3.3 --- Determination of phenolic compounds --- p.38
Chapter 2.3.3.1 --- Total phenolic content --- p.38
Chapter 2.3.3.2 --- Identification of phenolic acids --- p.39
Chapter 2.4 --- Summary --- p.45
Chapter Chapter 3: --- Anti-angiogenic properties of the Aa water extract --- p.46
Chapter 3.1 --- Introduction --- p.46
Chapter 3.1.1 --- Angiogenesis --- p.46
Chapter 3.1.1.1 --- Process of angiogenesis --- p.46
Chapter 3.1.1.2 --- Regulations of angiogenesis --- p.47
Chapter 3.1.1.2.1 --- Fibroblast growth factor (bFGF) --- p.47
Chapter 3.1.1.2.2 --- Vascular endothelial growth factor (VEGF) --- p.48
Chapter 3.1.2 --- Tumor angiogenesis --- p.49
Chapter 3.1.2.1 --- ROS generation in tumor cells --- p.50
Chapter 3.1.2.2 --- Hydrogen peroxide and VEGF --- p.51
Chapter 3.1.2.3 --- Previous studies on tumor angiogenesis --- p.52
Chapter 3.1.2.3.1 --- ROS and endothelial cells proliferation --- p.52
Chapter 3.1.2.3.2 --- VEGF and endothelial cells functions --- p.53
Chapter 3.1.3 --- Use of antioxidants in cancer treatment --- p.53
Chapter 3.1.3.1 --- Antioxidant use of cancer therapy --- p.53
Chapter 3.1.3.2 --- Antioxidant and endothelial cells functions --- p.54
Chapter 3.1.3.3 --- Anti-angiogenic effects of polyphenols --- p.56
Chapter 3.1.3.3.1 --- Phenolic acids --- p.56
Chapter 3.1.3.3.2 --- Tea catechin --- p.57
Chapter 3.1.3.3.3 --- Resveratrol --- p.57
Chapter 3.1.3.3.4 --- Genistein --- p.58
Chapter 3.2 --- Principles of Methods and Experimental Protocols --- p.60
Chapter 3.2.1 --- Sample preparation --- p.60
Chapter 3.2.2 --- Toxicity of the Aa water extract --- p.60
Chapter 3.2.2.1 --- Limulus amebocyte lysate (LAL) test --- p.60
Chapter 3.2.2.2 --- Toxicity towards normal cells --- p.61
Chapter 3.2.2.2.1 --- Cell line and its subculture --- p.61
Chapter 3.2.2.2.2 --- Colorimetric (MTT) assay --- p.62
Chapter 3.2.3 --- Effect of the Aa water extract on cancer cells --- p.63
Chapter 3.2.3.1 --- Cell line and its subculture --- p.63
Chapter 3.2.3.2 --- Redox status --- p.63
Chapter 3.2.3.3 --- VEGF secretion --- p.65
Chapter 3.2.4 --- In vitro cell culture anti-angioenesis analysis --- p.66
Chapter 3.2.4.1 --- Cell line and its subculture --- p.66
Chapter 3.2.4.2 --- Endothelial cells proliferation --- p.67
Chapter 3.2.4.3 --- Endothelial cells migration --- p.68
Chapter 3.2.4.3.1 --- Wound healing assay --- p.68
Chapter 3.2.4.3.2 --- Transwell culture insert assay --- p.69
Chapter 3.2.4.4 --- Endothelial cells tubule formation --- p.71
Chapter 3.2.5 --- In vitro organ culture anti-angiogenesis analysis --- p.72
Chapter 3.2.5.1 --- Aortic ring assay --- p.72
Chapter 3.2.6 --- Statistical analysis --- p.74
Chapter 3.3 --- Results and Discussions --- p.75
Chapter 3.3.1 --- Toxicity of the Aa water extract --- p.75
Chapter 3.3.1.1 --- Limulus amebocyte lysate (LAL) test --- p.75
Chapter 3.3.1.2 --- Toxicity towards normal cells --- p.75
Chapter 3.3.2 --- Effect of the Aa water extract on cancer cells --- p.77
Chapter 3.3.2.1 --- Redox status --- p.77
Chapter 3.3.2.2 --- VEGF secretion --- p.79
Chapter 3.3.2.3 --- Relationship between intracellular ROS and VEGF secretion detected --- p.80
Chapter 3.3.3 --- Effect of the Aa water extract on angiogenesis --- p.82
Chapter 3.3.3.1 --- Endothelial cells proliferation --- p.82
Chapter 3.3.3.2 --- Endothelial cells migration --- p.84
Chapter 3.3.3.2.1 --- Wound healing assay --- p.84
Chapter 3.3.3.2.2 --- Transwell culture insert assay --- p.87
Chapter 3.3.3.3 --- Endothelial cells tubule formation --- p.90
Chapter 3.3.3.4 --- Aortic ring assay --- p.97
Chapter 3.3.4 --- Effect of phenolic acids on endothelial cells --- p.101
Chapter 3.3.4.1 --- Endothelial cells proliferation --- p.101
Chapter 3.3.4.2 --- Endothelial cells migration --- p.102
Chapter 3.3.4.2.1 --- Wound healing assay --- p.102
Chapter 3.3.4.2.2 --- Transwell culture insert assay --- p.105
Chapter 3.3.4.3 --- Endothelial cells tubule formation --- p.106
Chapter 3.3.4.4 --- Aortic ring assay --- p.112
Chapter 3.4 --- Summary --- p.116
Chapter Chapter 4 --- Conclusions and future works --- p.118
References --- p.121
40
„Profiling of substrate-specificity and rational design of peptidomimetic inhibitors for 3C-like proteases of coronaviruses“. Thesis, 2010. http://library.cuhk.edu.hk/record=b6075034.
Annotation:
3C-like protease (3CLpro) of severe acute respiratory syndrome-coronavirus (SARS-CoV) is required for autoprocessing of the polyproteins 1a and 1ab, and is a potential target for treating coronaviral infection. To obtain a thorough understanding of its substrate preference, we created a substrate library of 19 x 8 variants by performing saturation mutagenesis on the autocleavage sequence at P5 to P3' positions. The substrate sequences were inserted between cyan and yellow fluorescent proteins so that the cleavage rates were monitored by in vitro fluorescence resonance energy transfer (FRET). The relative cleavage rate for different substrate sequences was correlated with various structural properties. P5 and P3 positions prefer residues with high beta-sheet propensity P4 prefers small hydrophobic residues: P2 prefers hydrophobic residues without beta-branch. Gln is the best residue at P1 position, but observable cleavage can be detected with His and Met substitutions. P1' position prefers small residues, while P2' and P3' positions have no strong preference on residue substitutions. Noteworthy, solvent exposed sites such as P5, P3 and P3' positions favour positively charged residues over negatively charged one, suggesting that electrostatic interactions may play a role in catalysis. A super-active substrate, which combined the preferred residues at P5 to P1 positions, was found to have 2.8 fold higher activity than the wild-type sequence.Inhibition of SARS-CoV 3CLpro proteolytic activity suppresses virion replication and virus-induced cytopathic effects. Peptidomimetic inhibitors with nitrile warheads, which inhibit Cys protease activity, have been applied for clinical therapy. To investigate whether the nitrile group can target 3CLpro, a series of nitrile-based peptidomimetic inhibitors with various protective groups, peptide length and peptide sequences were synthesized. Inhibitor potency in terms of IC50 and Ki values was determined by FRET assay. Most of these nitrile-based inhibitors in micromolar range can significantly reduce 3CLpro activity. The most potent inhibitor is the tetrapeptidomimetie inhibitor linked with carbobenzyloxy (cbz) group 'cbz-AVLQ-CN' with IC50 and Ki values of 5.9 +/- 0.6 muM and 0.62 +/- 0.11 muM respectively. Crystal structures of 3CLpro-inhibitor complexes demonstrated that nitrite warhead covalently bonded to Cys145, while P1 -- P4 residues interacted with 3CLpro as substrate bound. The cbz group in 'cbz-AVLQ-CN' flipped into a cavity of Gu166 -- Pro168, providing an extra binding force to enhance inhibitor potency. In conclusion, the nitrile-based peptidomimetic inhibitor with cbz group is a convincing model for drug development.
Substrate specificities of various 3CLpro were further investigated by using the substrate library of SARS-CoV 3CLpro. Among various viral strains, the proteases of HCoV-NL63, HCoV-OC43 and infectious bronchitis virus (IBV) were selected from group I, IIa and III respectively for specificity profiling. Their proteolytic rates against 19 x 8 variants were obtained by FRET assay, and correlated with structural properties of substituting residues. Like SARS-CoV 3CLpro in group IIb, these 3CLpro consistently prefer small hydrophobic P4 residues, positively charged P3 residues, hydrophobic P2 residues without beta-branch, P1-Gln and small P1' residues. These proteases also tend to accommodate P5 and P3' residues with positive charge, and P2' residues with small size. In contrast, their preferences on secondary structure are diverse. Correlation was found between IBV 3Clpro activity and beta-sheet propensity at P5 position, while no strong correlation with secondary structure propensities was observed in HCoV-NL63 and HCoV-0C43. Collectively, all 3CLpro share universal preferences on charge, side chain volume and hydrophobicity, but not secondary structure. Their relative activities against universal and specific super-active substrates were elevated to 1.4 -- 4.3, showing synergetic effects by combining preferred residues. These substrates were examined by group I HCoV-229E and group IIa HCoV-HKU1 in parallel. Their activities were highly comparable to those of other group members.
Chuck, Chi Pang.
Adviser: Chi-Cheong Wan.
Source: Dissertation Abstracts International, Volume: 73-02, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves [179]-187).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
41
„Growth inhibitory effect of docosahexaenoic acid on human melanoma A375 cells“. 2007. http://library.cuhk.edu.hk/record=b5896766.
Annotation:
Tong, Kit Fong.Thesis (M.Phil.)--Chinese University of Hong Kong, 2007.
Includes bibliographical references (leaves 91-104).
Abstracts in English and Chinese.
Abstract --- p.i
Acknowledgements --- p.vi
Table of Contents --- p.vii
List of Figures --- p.x
List of Tables --- p.xii
List of Abbreviations --- p.xiii
Chapter Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Cancer --- p.2
Chapter 1.1.1 --- Tumor development --- p.2
Chapter 1.1.2 --- Cell cycle --- p.4
Chapter 1.1.3 --- Apoptosis --- p.9
Chapter 1.1.3.1 --- The extrinsic pathway --- p.14
Chapter 1.1.3.2 --- The intrinsic pathway --- p.16
Chapter 1.1.3.3 --- The Bcl-2 family proteins --- p.17
Chapter 1.1.3.4 --- Execution of apoptosis --- p.20
Chapter 1.1.4 --- Melanoma --- p.22
Chapter 1.2 --- Polyunsaturated fatty acids (PUFAs) --- p.24
Chapter 1.2.1 --- "Chemistry, classification, metabolic conversion and sources …" --- p.24
Chapter 1.2.2 --- Epidemiology studies --- p.27
Chapter 1.2.3 --- Docosahexaenoic acid (DHA) --- p.28
Chapter 1.2.3.1 --- Sources --- p.28
Chapter 1.2.3.2 --- DHA and cancer --- p.29
Chapter 1.3 --- Objectives --- p.33
Chapter Chapter 2 --- Materials and Methods --- p.34
Chapter 2.1 --- In vitro studies of DHA on growth and survival of human cancer cells --- p.34
Chapter 2.1.1 --- Cell cultures --- p.34
Chapter 2.1.2 --- Studies of growth inhibition of DHA on human cancer cells --- p.35
Chapter 2.1.2.1 --- MTT assay --- p.35
Chapter 2.1.2.2 --- Chemiluminescent-bromodeoxyuridine (Chemi-BrdU) immunoassay --- p.36
Chapter 2.1.3 --- Studies of growth inhibitory mechanism of DHA on A375 cells. --- p.38
Chapter 2.1.3.1 --- DNA -flow cytometry analysis --- p.38
Chapter 2.1.3.2 --- Western blot analysis --- p.39
Chapter 2.1.3.3 --- Caspase inhibitor studies --- p.42
Chapter 2.1.3.4 --- Mitochondrial membrane potential analysis --- p.42
Chapter 2.2 --- In vivo study of the anticancer effect of DHA on A375 cells --- p.44
Chapter 2.2.1 --- Animals --- p.44
Chapter 2.2.2 --- Cell inoculation and treatments --- p.44
Chapter 2.2.3 --- Western blot analysis --- p.45
Chapter 2.3 --- Statistical analysis --- p.46
Chapter Chapter 3 --- Results --- p.47
Chapter 3.1 --- In vitro studies of DHA on growth and survival of human canccr cells --- p.47
Chapter 3.1.1 --- DHA reduced proliferation and survival of human cancer cells --- p.47
Chapter 3.1.2 --- DHA modulated cell cycle of A375 cells --- p.52
Chapter 3.1.3 --- DHA induced apoptosis in A375 cells --- p.55
Chapter 3.1.4 --- Caspase activations were involved in the DHA-induced apoptosis in A375 cells --- p.59
Chapter 3.1.5 --- "Caspase 3´ة 6, 8 and 9 were activated in DHA-induced apoptosis of A375 cells" --- p.62
Chapter 3.1.6 --- DHA dissipated mitochondrial membrane potential in A375 cells --- p.66
Chapter 3.1.7 --- DHA triggered the mitochondrial pathway of apoptosis --- p.68
Chapter 3.1.8 --- DHA triggered the death receptor pathway of apoptosis --- p.71
Chapter 3.2 --- In vivo study of the anticancer effect of DHA on A375 cells --- p.74
Chapter 3.2.1 --- Effect of DHA on the growth ofA375 xenograft in athymic Bαlb/c mice --- p.74
Chapter 3.2.2 --- DR4 and TRAIL were upregulated by DHA treatment in A375 solid tumor --- p.77
Chapter Chapter 4 --- Discussion --- p.79
References --- p.91
42
„Anti-angiogenic effects and mechanisms of the Chinese herbs rhizoma rhei, fructus alpiniae and rhizoma kaempferiae“. Thesis, 2010. http://library.cuhk.edu.hk/record=b6075075.
Annotation:
All the results showed that TCMs can provide a source for discovering anti-angiogenic agents for the treatment of cancer, and all these experiments in the zebrafish and mammalian models further confirmed the value of zebrafish model in anti-angiogenic drug discovery.Angiogenesis refers to the formation of new blood capillaries from pre-existing ones, and is essential in a series of normal physiological processes such as embryonic development and pathological responses. However, persistent unregulated angiogenesis causes "angiogenic diseases" such as diabetic retinopathy, tumor growth and metastasis, rheumatoid arthritis, and inflammatory diseases. The linkage between angiogenesis, tumor growth and metastasis was first hypothesized by Dr. Judah Folkman in the 1970s, and now this controversial idea is widely accepted and the inhibition of angiogenesis, or anti-angiogenesis, is considered as a promising anticancer therapeutic strategy. Bevacizumab (Avastin RTM by Genentech Inc.), the first approved anti-angiogenic drug by U.S. FDA in 2004, is a humanized monoclonal antibody to inhibit endothelial cell proliferation and angiogenesis for the treatment of metastatic colorectal cancer, non-small cell lung cancer, advanced breast cancer, glioblastoma, metastatic renal cell cancer.
Anti-angiogenic therapy in cancer treatment has led to the development of compounds designed to control a tumor's growth by blocking its ability to develop a blood supply. The development of agents with different mechanisms of action requires powerful preclinical models for the analysis and optimization of the therapy. Some in vitro and in vivo anti-angiogenic assays are already developed, for example, Human Umbilical Vein Endothelial Cell (HUVEC) assay, Chorioallantoic Membrane assay, Matrigel plug assay et al. Zebrafish, as a relatively new model organism, is firmly established as a powerful research platform for many areas of biology and drug discovery, allowing the testing of bioactive compounds in a whole organism and in cells undergoing normal cell-cell and cell-matrix interactions. Many anti- and pro-angiogenic molecules tested in zebrafish demonstrated similar effects to those observed in humans or other mammalian models. Besides providing a powerful platform for drug screening, zebrafish model can also be used for probing biological processes, and generate insights into mechanisms.
Cancer is a generic term for a large group of diseases that can affect any part of the body, which causes a vast medical problem and is a leading cause of death worldwide nowadays. However, for many years the main methods of treating cancer have been surgery, radiotherapy and chemotherapy. Among these treatments, chemotherapy has played a major role in cancer therapy for half a century. Despite improving managements and efforts, it is not surprising that the prognosis has not greatly improved because of the limitations of current therapies, such as toxicity, inherent and acquired resistance, and metastatic spread. This calls for novel cancer therapies and new group of anticancer agents for selectively targeting cancers without or with lower toxicity to normal tissues.
Traditional Chinese medicines (TCMs) have long been recognized as a rich source for discovering drugs, and various TCMs and their components have shown anti-angiogenic properties. In this thesis study, as a continuing pursuit for elucidating the anti-angiogenic properties of TCMs, our attention is focused on those with effects of anti-inflammation, anti-rheumatoid arthritis and anti-cancer. On zebrafish screening model, three of the selected TCMs, Rheum palmatum, Alpinia oxyphylla (seeds), and Kaempferia galanga showed potential anti-angiogenic activity, indicating the existence of potent anti-angiogenic components in these herbs. The ethyl acetate fraction of R. palmatum showed strong inhibition of vessel formation in zebrafish embryos. Further testing of the anthraquinones of this herb showed three of them displayed potent anti-angiogenic activities. The most potent compound---rhein could inhibit HUVEC migration and affect the mRNA expression of vegfa, kdr, angiopoietin1/2 and tie1/2; The n-hexane and ethyl acetate fractions of A. oxyphylla and K. galangal showed anti-angiogenic potentials both in zebrafish and HUVEC assays. The n-hexane and ethyl acetate fractions of A. oxyphylla could both inhibit the proliferation, migration and tube formation processes of HUVEC. And the most potential component, trans-ethyl-p-methoxycinnamate from K. galanga, could inhibit HUVEC migration and tube formation, and reduce all gene expressions involved in angiogenesis process except for vegfa.
He, Zhiheng.
Adviser: Wei Ge.
Source: Dissertation Abstracts International, Volume: 73-02, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 87-108).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
43
„Investigating the chemopreventive effect of hesperetin, luteolin and cyclooxygenase inhibitors in a mouse model of breast cancer“. 2012. http://library.cuhk.edu.hk/record=b5549528.
Annotation:
乳腺癌是女性最常見的腫瘤之一，多發生在女性絶經後，並具有雌激素依賴性。芳香化酶(CYP19)是雌激素生物合成過程中的關鍵酶，而芳香化酶抑製劑(AI)則被用於替代治療雌激素依賴性的乳腺癌。然而，AI在降低雌激素水平的同時能夠引起骨質酥鬆。此項研究的目的是找尋AI替代物。黃酮類化合物是一種多酚化合物，廣泛分佈于植物中。我們先前的研究發現二氢黄酮陈皮素能夠抑制芳香化酶的生物活性，并且抑制芳香化酶高表達的乳腺癌生長。在本研究中，我們發現陳皮素在抑制腫瘤生長的同時能夠降低来曲唑引起的骨質流失。木犀草素是另外一種黄酮类化合物，它同樣能夠抑制芳香化酶的活性并減少骨流失。而與陳皮素不同的是，它能夠抑制芳香化酶的表達。在芳香化酶高表達的乳腺癌細胞(MCF-7 aro)中，木犀草素抑制芳香化酶活性的IC50是3 μM。在MCF-7 細胞中，5 μM的木犀草素能夠抑制CYP19 mRNA 的表達，螢光素酶報告實驗顯示木犀草素是通過作用于啟動子I.3和II來抑制CYP19的表達。蛋白印跡實驗表明木犀草素抑制CYP19表達的分子機制可能通過調節JNK信號通路進而減少AP-1的活性來實現。動物實驗結果顯示木犀草素能夠抑制MCF-7aro腫瘤的生長并改善來曲唑引起的骨流失。
環氧化酶(COX)是花生四烯酸轉化為前列腺素途徑中的一種關鍵酶。研究發現COX-2在乳腺癌組織中廣泛表達。本實驗研究了COX抑製劑在裸鼠動物模型中對乳腺癌腫瘤的作用機制。研究結果表明塞來昔布和阿司匹林在不影響血液中雌激素水平的情況下抑制乳腺癌腫瘤的生長。蛋白印迹實驗顯示這兩種藥物能夠降低腫瘤中COX-2，Cyclin A和Bcl-xL的表達。miR-98, miR-222和miR-145也能夠被塞來昔布和阿司匹林影響。
本研究表明陳皮素，木犀草素及COX抑制劑有潛力成為替代AI的化學治療藥物或共同治療藥物。
Breast cancer is one of the most prevalent cancers affecting women. The majority of breast tumor growth occurred in the post-menopausal period are estrogen dependent. Aromatase (CYP19) catalyzes the rate-limiting step in the synthetic reaction of estrogen and aromatase inhibitors (AIs) are contemporary treatment for estrogen-positive breast cancer. However, estrogen-lowering drugs may promote osteoporosis. Our objective of this study further identified some alternatives for AIs.
Flavonoids are polyphenolic compounds that are ubiquitously distributed in plants. We have previously found that the flavanone hesperetin can inhibit the activity of aromatase and suppress aromatase-expressing breast tumor growth. In this project, we investigated the potential interaction between hesperetin and the AI letrozole in a mouse model. Our results showed that hesperetin could inhibit the tumor growth and reduce bone loss induced by letrozole. Similarly, another flavonoid luteolin also inhibited aromatase and prevented bone deterioration as observed in this project. In cells stably transfected with CYP19 (MCF-7aro), luteolin inhibited the aromatase activity with an IC50 value of 3μM. In addition, 5μM luteolin significantly reduced CYP19 mRNA expression in MCF-7 cells. Luciferase reporter assay revealed that luteolin could suppress CYP19 transcription at promoter regions I.3 and II. Western analysis illustrated that JNK signaling pathway was involved and deactivation of AP-1 could be the underlying molecular mechanism. Subsequently, we examined the effect in vivo. Our results showed that luteolin could inhibit the MCF-7aro tumor growth and improved bone loss induced by letrozole.
Cyclooxygenase (COX) is an enzyme responsible for the conversion of arachidonic acid into prostaglandins. It is over-expressed in breast cancer tissue and an increased expression of COX-2 was also observed in the xenograft model employed in this project. In the last study we evaluated the importance of COX-2 in breast tumor growth in this model. Our data showed that celecoxib and aspirin could significantly suppress the tumor growth without changing the plasma estrogen level. Western analysis illustrated that COX-2, Cyclin A, Bcl-xL and ER were reduced in celecoxib- and aspirin- treated tumor samples and miR-98, miR-222 and miR-145 were altered by celecoxib or aspirin.
After all, this project demonstrated that hesperetin, luteolin and COX-inhibitors could be potential chemopreventive or co-therapeutic agents.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Li, Fengjuan.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2012.
Includes bibliographical references (leaves 131-148).
Abstract also in Chinese.
ACKNOWLEDGEMENTS --- p.I
ABSTRACT --- p.II
摘要 --- p.IV
LIST OF ABBREVIATIONS --- p.V
TABLE OF CONTENTS --- p.VII
CHAPTER 1 --- p.1
GENERAL INTRODUCTION --- p.1
Chapter 1.1 --- Types of Breast Cancer --- p.3
Chapter 1.2 --- Nuclear Receptor Signaling Pathways in Breast Cancer --- p.5
Chapter 1.3 --- Estrogen and Breast Cancer --- p.7
Chapter 1.4 --- Estrogen and Bone Health --- p.8
Chapter 1.5 --- Estrogen Biosynthesis and Aromatase --- p.10
Chapter 1.6 --- Tissue Specific Promoter for Aromatase Expression --- p.13
Chapter 1.7 --- Nuclear Receptors and Aromatase Promoter Regulation --- p.15
Chapter 1.8 --- Signaling Pathway and Aromatase Expression --- p.17
Chapter 1.9 --- Cell Cycle in Breast Cancer --- p.20
Chapter 1.10 --- Cell Apoptosis --- p.23
Chapter 1.11 --- Treatment of breast cancer --- p.25
Chapter 1.12 --- Phytoestrogens --- p.29
Chapter 1.13 --- Aim of My Study --- p.32
CHAPTER 2 --- p.33
MATERIALS AND METHODS --- p.33
Chapter 2.1 --- Chemicals and Materials --- p.33
Chapter 2.1.1 --- Chemicals --- p.33
Chapter 2.1.2 --- Plasmids --- p.33
Chapter 2.2 --- Cell Culture --- p.33
Chapter 2.3 --- Aromatase Activity Assay --- p.34
Chapter 2.4 --- Quantitative Real Time PCR --- p.36
Chapter 2.4.1 --- RNA Isolation and cDNA Synthesis --- p.36
Chapter 2.4.2 --- Quantitative Real Time PCR Assay --- p.37
Chapter 2.4.3 --- MiRNA Quantitative Real Time PCR Assay --- p.38
Chapter 2.5 --- Western Blot --- p.39
Chapter 2.6 --- Measurement of Promoter Activity --- p.41
Chapter 2.6.1 --- Plasmid Preparation --- p.41
Chapter 2.6.2 --- Transient Transfection and Dual-Luciferase Assay --- p.42
Chapter 2.7 --- Electrophoretic Mobility Shift Assay (EMSA) --- p.43
Chapter 2.7.1 --- Nuclear protein extraction --- p.43
Chapter 2.7.2 --- Electrophorectic Mobility Shift Assay --- p.44
Chapter 2.8 --- Animal Experiment Design --- p.45
Chapter 2.8.1 --- Animal Model for Hesperetin Study --- p.45
Chapter 2.8.2 --- Animal Model for Luteolin Study --- p.46
Chapter 2.8.3 --- Animal Model for Cycooxygenase Inhibitors Study --- p.48
Chapter 2.8.4 --- Serum Estradiol Determination --- p.49
Chapter 2.8.5 --- Analysis of serum lipoproteins --- p.49
Chapter 2.8.6 --- Bone Image Acquisition and Region of Interest Selection --- p.50
Chapter 2.9 --- Statistical Analysis --- p.50
CHAPTER 3 --- p.51
The citrus flavonone hesperetin prevents letrozole- induced bone loss in a mouse model of breast cancer --- p.51
Chapter 3.1 --- Introduction --- p.51
Chapter 3.2 --- Results --- p.54
Chapter 3.2.1 --- Murine Body Weight and Liver Weight --- p.54
Chapter 3.2.2 --- Effect of Hesperetin and Letrozole on Xenograft Growth in Ovariectomized Mice --- p.55
Chapter 3.2.3 --- Hesperetin Reduced Plasma Estradiol Concentration --- p.58
Chapter 3.2.4 --- PS2 mRNA Expression in Tumor --- p.59
Chapter 3.2.5 --- Uterine Wet Weight --- p.60
Chapter 3.2.6 --- Hesperetin Prevent Bone Deterioration Induced by Letrozole --- p.61
Chapter 3.3 --- DISCUSSION --- p.63
CHAPTER 4 --- p.66
dIETARY FLAVONOID LUTEOLIN ON cyp19 transcription in the breast cancer cells mcf-7 --- p.66
Chapter 4.1 --- Introduction --- p.66
Chapter 4.2 --- Results --- p.68
Chapter 4.2.1 --- Inhibitory Effect of Luteolin on Aromatase Activity --- p.68
Chapter 4.2.2 --- Luteolin Reduced Aromatase mRNA Expression in MCF-7 Cells --- p.70
Chapter 4.2.3 --- Effect of Luteolin on Promoter I.3/II Activity of CYP19 in MCF-7 Cells --- p.71
Chapter 4.2.4 --- The Effect of Luteolin on Truncation CYP19 Gene Reporter Assay --- p.72
Chapter 4.2.5 --- Luteolin Reduced AP-1 Binding in Promoter I.3/II DNA Fragment --- p.74
Chapter 4.2.6 --- Inhibitory Effect of Luteolin on Protein Kinase Signaling --- p.76
Chapter 4.3 --- Discussion --- p.78
CHAPTER 5 --- p.83
interaction OF LUTEOLIN and letrozole in a postmenopausal breast cancer model --- p.83
Chapter 5.1 --- Introduction --- p.83
Chapter 5.2 --- Results --- p.86
Chapter 5.2.1 --- Luteolin and letrozole treatment had no effect on mouse body weight and liver weight --- p.86
Chapter 5.2.2 --- Effect of luteolin and Letrozole on Xenograft Growth in Ovariectomized Mice --- p.88
Chapter 5.2.3 --- Luteolin reduced plasma estradiol concentration --- p.91
Chapter 5.2.4 --- Luteolin Counteracted Uterine Weight Reduction under Letrozole Treatment --- p.92
Chapter 5.2.5 --- Luteolin Prevented Bone Deterioration Induced by Letrozole --- p.93
Chapter 5.2.6 --- The Effect of Luteolin on Plasma TC and TG --- p.95
Chapter 5.2.7 --- Luteolin Increased HDL Level and Reduced the Ratio of LDL/HDL --- p.97
Chapter 5.2.8 --- Effect of Luteolin on Cell Cycle and Apoptotic Protein Expression --- p.99
Chapter 5.3 --- DISCUSSION --- p.104
CHAPTER 6 --- p.107
cyclooxygenase inhibitors suppresse breast tumor growth in NUDE MICE --- p.107
Chapter 6.1 --- Introduction --- p.107
Chapter 6.2 --- Results --- p.109
Chapter 6.2.1 --- Celecoxib and aspirin treatment had no effect on mouse body weight and liver weight --- p.109
Chapter 6.2.2 --- Effect of celecoxib and aspirin on Xenograft Growth in Ovariectomized Mice --- p.111
Chapter 6.2.3 --- Celecoxib and aspirin had no effect on plasma estradiol concentration --- p.113
Chapter 6.2.4 --- Celecoxib and Aspirin Had no Effect on Uterine Weight --- p.114
Chapter 6.2.5 --- Protein expression of COX-2, Cell cycle-related and cell Apoptotic Genes --- p.115
Chapter 6.2.6 --- Detection of Related miRNA Expression Level in Tumors --- p.118
Chapter 6.2.7 --- c-Myc mRNA Expression Level were Regulated in Tumors --- p.121
Chapter 6.3 --- DISCUSSION --- p.124
CHAPTER 7 --- p.127
SUMMARY --- p.127
REFERENCE --- p.131
44
Robarge, Jason Dennis. „Aromatase inhibitors produce hypersensitivity in experimental models of pain : studies in vivo and in isolated sensory neurons“. Thesis, 2014. http://hdl.handle.net/1805/6056.
Annotation:
Indiana University-Purdue University Indianapolis (IUPUI)Aromatase inhibitors (AIs) are the current standard of care for the treatment of hormone receptor positive breast cancer in postmenopausal women. Nearly one-half of patients receiving AI therapy develop musculoskeletal toxicity that is characterized by joint and/or muscle pain and approximately one-fourth of patients discontinue their therapy as a result of musculoskeletal pain. Since there are no effective strategies for prevention or treatment, insight into the mechanisms of AI-induced pain is critical to improve treatment. However, there are few studies of AI effects in animal models of nociception. To determine whether AIs produce hypersensitivity in animal models of pain, I examined the effects of AI administration on mechanical, thermal, and chemical sensitivity in rats. The results demonstrate that (1) repeated injection of 5 mg/kg letrozole in male rats produces mechanical, but not thermal, hypersensitivity that extinguishes when drug dosing is stopped; (2) administering a single dose of 1 or 5 mg/kg letrozole in ovariectomized (OVX) rats also induces mechanical hypersensitivity, without altering thermal sensitivity and (3) a single dose of 5 mg/kg letrozole or daily dosing of letrozole or exemestane in male rats augments flinching behavior induced by intraplantar ATP injection. To determine whether the effects of AIs on nociceptive behaviors are mediated by activation or sensitization of peptidergic sensory neurons, I determined whether letrozole exposure alters release of calcitonin gene-related peptide (CGRP) from isolated rat sensory neurons and from sensory nerve endings in rat spinal cord slices. No changes in basal, capsaicin-evoked or high extracellular potassium-evoked CGRP release were observed in sensory neuronal cultures acutely or chronically exposed to letrozole. Furthermore, letrozole exposure did not alter the ability of ATP to augment CGRP release from sensory neurons in culture. Finally, chronic letrozole treatment did not augment neuropeptide release from spinal cord slices. Taken together, these results do not support altered release of this neuropeptide into the spinal cord as mediator of letrozole-induced mechanical hypersensitivity and suggest the involvement of other mechanisms. Results from this dissertation provide a new experimental model for AI-induced hypersensitivity that could be beneficial in delineating mechanisms mediating pain during AI therapy.
45
„Search of inhibitors that target HIV pre-mRNA splicing to overcome drug resistance“. 2012. http://library.cuhk.edu.hk/record=b5549184.
Annotation:
引發獲得性免疫缺陷綜合癥（AIDS）的人類免疫缺陷病毒（HIV）是一種逆轉錄病毒。過去的十餘年間，高效抗逆轉錄病毒治療療法（HARRT），在抗病毒感染方面取得了很大的成功。高效抗逆轉錄病毒治療療法是一種將多種抗逆轉錄病毒藥物複合的藥物聯用療法。然而，因為病毒的逆轉錄過程極易突變，導致HIV已經可以對大多數使用的抑製藥物產生抗藥性。因此，有越來越多的需要去尋找新型的抗病毒複製機理，例如將人體細胞蛋白作為載體，來達到克服病毒抗藥性的目的。HIV-1的複製離不開宿主細胞的剪接因子，例如SR蛋白。選擇性剪接因子ASF/SF2，一個典型的調控pre-RNA剪接的SR蛋白，在HIV-1的pre-mRNA剪接和複製中起到了很重要的調控作用。ASF/SF2和其他SR蛋白一樣，都被丝氨酸/苏氨酸蛋白激酶（SRPK）磷酸化，磷酸化位點位於C端的丝氨酸/苏氨酸結構域（RS domain）。SRPK通過磷酸化來調節ASF/SF2在細胞中的分佈。對於SRPK 和ASF/SF2複合物的結構學和功能學研究指出，ASF/SF2的docking motif和SRPK1的遠離活性位點的docking groove存在很強的相互作用。而這種相互作用是調節磷酸化過程關鍵。所以，在我們的研究過程中，我們希望通過阻斷2個蛋白的相互作用來干擾ASF/SF2的磷酸化，進而抑制其在HIV-1 pre-mRNA剪接過程中的活性。
我們採用以結構為基礎的藥物模擬篩選，來選擇潛在的抑制物，達到通過抑制物與docking groove的相互作用來阻斷ASF/SF2和SRPK1的相互作用，以達到抑制磷酸化的目的。我們使用的數據庫來自于ZINC數據庫（UCSF），包括天然產物數據庫和SPECS。我們採用AutoDock Vina 和AutoDock 4.2 二個模擬軟件來栓選數據庫中351473个化合物。并從中選出50個潛在的化合物用作之後的化學生物學測試。體外的激酶活性試驗顯示，6個化合物對ASF/SF2的磷酸化有抑製作用。
體外的HIV-1 pre-mRNA剪接實驗顯示，5個化合物在逆轉錄PCR（RT-PCR）中有一定得抑制效果。和DMSO對照組相比，在抑製劑作用下剪接產物的生成被抑制。HIV-1病毒合胞體感染實驗顯示，有一個化合物對病毒的感染起到了一定的抑制作用。
其他的測試實驗還在進行中，包括對SRPK1和抑制物複合物的結構研究，從而更好的研究抑制物的作用機理。以及，採用表面等離子共振波譜來進行動力學研究和其他關於化合物在病毒複製過程中的實驗測試。
Human immunodeficient virus (HIV) is a retrovirus that cause acquired immunodeficiency syndrome (AIDS). Highly active antiretroviral therapy (HAART) is a treatment of HIV infection that uses combinations of antiretroviral drugs and has achieved great success in the past two decades. However, since the reverse transcription process of viral RNA is notoriously prone to error, HIV-1 can acquire resistance to nearly all known inhibitors and has started to develop resistance to HAART. Therefore, there is an ongoing search for new drugs with novel inhibitory mechanism such as targeting cellular proteins essential for HIV-1 replication to overcome drug resistance of the virus.
HIV-1 mRNA undergoes complex splicing and the expression of the integrated HIV-1 provirus is largely dependent on the host’s splicing machinery which assembly requires splicing factors such as serine-arginine rich proteins (SR proteins). Alternative splicing factor/splicing factor 2 (ASF/SF2), a prototypic SR protein that is essential for pre-mRNA splicing, has been shown to play critical roles during HIV-1 pre-mRNA splicing and replication. ASF/SF2, like other SR proteins, is phosphorylated by SR protein-specific kinases (SRPKs) at its C-terminal arginine/serine (RS) domain, which governs its localization and metabolism. Structural and functional studies of SRPK1 in complex with ASF/SF2 has revealed that a docking groove on SRPK1 that is distal to the active site interacts strongly with a docking motif and the RS domain of ASF/SF2, leading to high affinity binding as well as regulating the mechanism of phosphorylation. In this study, we propose that by blocking this interaction, we might interfere the phosphorylation of ASF/SF2 and inhibit its activity during splicing of HIV-1 pre-mRNA.
Structure-based in silico screening method is adopted to identify potential inhibitors that bind to the docking groove of SRPK1 to block the binding and phosphorylation of ASF/SF2. The compound libraries being used include the Natual Products Database and SPECS database from ZINC (UCSF). 351,473 compounds have been screened using the program Autodock Vina as well as Autodock 4.0. Until now 50 potential candidates of inhibitor have been selected for biochemical analyses. In vitro kinase assays showed that six compounds exhibit inhibitory activity against the phosphorylation of ASF/SF2.
To test the effect of the selected inhibitors on the splicing of HIV-1 mRNA, ex vivo splicing assay has been performed. Current results showed that the synthesis of splicing products extracted from drug-treated cells was less efficient when compared to untreated cells. Biological assays testing the inhibitory effects of the compounds on viral infection are currently underway. Our preliminary result suggested that one of the compounds could indeed inhibit HIV-1 viral infection.
Other biochemical and biological analyses including structural study of kinase-inhibitor complexes to understand the mode of inhibition; measurement of binding kinetics using surface plasmon resonance spectroscopy (SPR); and biological assays testing the inhibitory effects of the compounds on replication are underway.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Yu, Xiyao.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2012.
Includes bibliographical references (leaves 95-107).
Abstracts also in Chinese.
Abstract --- p.I
摘要 --- p.III
Acknowledgements --- p.V
TABLE OF CONTENTS --- p.VI
LIST OF FIGURES --- p.IX
LIST OF TABLES --- p.XI
Chapter Chapter I --- : Introduction --- p.1
Chapter 1.1 --- HIV, HAART and HIV Drug Resistance --- p.2
Chapter 1.2 --- HIV-1 alternative splicing mechanism --- p.9
Chapter 1.3 --- SR Protein Family --- p.13
Chapter 1.4 --- Functional roles of SR protein in HIV pre-mRNA splicing --- p.16
Chapter 1.5 --- Phosphorylation States of SR Proteins --- p.18
Chapter 1.6 --- SR protein Kinase --- p.20
Chapter 1.7 --- Interaction between SRPK1 and ASF/SF2 --- p.23
Chapter 1.8 --- IDC16 and SPRIN340 --- p.26
Chapter 1.9 --- Structure-based drug screening --- p.27
Chapter 1.10 --- AutoDock Suite --- p.29
Chapter 1.11 --- Kinase-substrate interaction inhibitors --- p.30
Chapter 1.12 --- Focus of study --- p.34
Chapter Chapter II --- : Materials and Methods --- p.35
Chapter 2.1 --- Materials --- p.36
Chapter 2.1.1 --- Bacterial strain --- p.36
Chapter 2.1.2 --- Antibodies --- p.36
Chapter 2.1.3 --- Cell line --- p.36
Chapter 2.1.4 --- Plasmid --- p.36
Chapter 2.1.5 --- Reagents --- p.38
Chapter 2.2 --- Expression and purification of Recombinant protein --- p.38
Chapter 2.3 --- In silico screening of inhibitors --- p.44
Chapter 2.4 --- Kinase Glo Assay --- p.45
Chapter 2.5 --- In vitro kinase assay --- p.45
Chapter 2.6 --- Cell Culture --- p.46
Chapter 2.7 --- MTT Assay --- p.46
Chapter 2.8 --- Immunocytochemistry --- p.47
Chapter 2.9 --- Ex vivo splicing assay --- p.47
Chapter 2.10 --- Surface plasmon resonance spectroscope --- p.48
Chapter Chapter III --- : Results --- p.50
Chapter 3.1 --- In silico screening of inhibitors --- p.51
Chapter 3.2 --- Selected Compounds Inhibits SRPK1 in Vitro --- p.60
Chapter 3.2.1 --- Protein purification --- p.60
Chapter 3.2.2 --- Inhibits ASF/SF2 Phosphorylation by SRPK --- p.66
Chapter 3.3 --- Surface Plasmon Resonance Binding Competition Assay --- p.76
Chapter 3.4 --- Inhibitors Alters HIV-1 Alternative Splicing ex Vivo --- p.79
Chapter 3.5 --- Cytotoxic effect of candidate compound on HeLa cells --- p.84
Chapter 3.6 --- Nature compound alters ASF/SF2 localization --- p.86
Chapter Chapter IV --- : Discussion and Conclusion --- p.89
References --- p.95
46
Chou, Jennie Yu. „Relationships among resident, physician, and facility characteristics, angiotensin-converting enzyme inhibitor use, and hospital utilization in elderly nursing home residents with heart failure“. Thesis, 2005. http://hdl.handle.net/2152/1846.
47
„Studies on the mechanisms and anti-tumor activities of green tea epicatechin isomers“. 2000. http://library.cuhk.edu.hk/record=b5890404.
Annotation:
by Ip Wai-Ki.Thesis (M.Phil.)--Chinese University of Hong Kong, 2000.
Includes bibliographical references (leaves 213-233).
Abstracts in English and Chinese.
ACKNOWLEDGEMENTS --- p.i
ABBREVIATIONS --- p.ii
ABSTRACT --- p.vi
撮要 --- p.x
TABLE OF CONTENTS --- p.xiv
Chapter CHAPTER 1: --- GENERAL INTRODUCTION
Chapter 1.1 --- Hematopoiesis --- p.1
Chapter 1.1.1 --- Introduction to Hematopoiesis --- p.1
Chapter 1.1.2 --- Cytokines in Hematopoiesis --- p.4
Chapter 1.2 --- Leukemia --- p.6
Chapter 1.2.1 --- Leukemia: Abnormalities in Blood Cell Formation --- p.6
Chapter 1.2.2 --- Classification of Leukemia --- p.8
Chapter 1.2.3 --- The Causes and Molecular Basis of Leukemia --- p.8
Chapter 1.2.4 --- Therapy of Leukemia --- p.11
Chapter 1.2.5 --- Control of Leukemia by Hematopoietic Growth Factors and Other Compounds --- p.12
Chapter 1.2.6 --- Molecular Control of Apoptosis and Cell Cycle in Leukemia --- p.13
Chapter 1.2.6.1 --- Regulation of Cell Cycle and Apoptosis by Genes and Regulatory Proteins --- p.14
Chapter 1.2.6.1.1 --- Cell Cycle --- p.14
Chapter 1.2.6.1.2 --- Apoptosis --- p.15
Chapter 1.2.6.2 --- Role of Apoptosis and Cell Cycle in the Development of Leukemia --- p.17
Chapter 1.3 --- Green Tea --- p.19
Chapter 1.3.1 --- Origin and Cultivation of Tea Plants --- p.19
Chapter 1.3.2 --- Classification and Manufacturing of Tea --- p.21
Chapter 1.3.3 --- The Chemistry of Tea --- p.22
Chapter 1.3.3.1 --- Chemical Composition of Tea --- p.22
Chapter 1.3.3.2 --- Separation and Purification of Green Tea Polyphenols --- p.27
Chapter 1.3.3.3 --- The Chemical Properties of Green Tea Polyphenols --- p.28
Chapter 1.3.4 --- Bioavailability and Pharmacokinetic of Green Tea Epicatechins --- p.28
Chapter 1.3.4.1 --- Human Studies --- p.29
Chapter 1.3.4.2 --- Animal Studies --- p.30
Chapter 1.3.5 --- Physiological and Pharmacological Activities of Green Tea Catechins --- p.31
Chapter 1.3.5.1 --- Anti-oxidative Activity --- p.32
Chapter 1.3.5.2 --- Hypocholesterolemic and Hypolipidemic Activity --- p.33
Chapter 1.3.5.3 --- Anti-inflammatory Activity --- p.34
Chapter 1.3.5.4 --- Anti-microbial Activity --- p.35
Chapter 1.3.5.5 --- Anti-mutagenic Activity --- p.36
Chapter 1.3.5.6 --- Anti-carcinogenesis --- p.37
Chapter 1.3.5.7 --- Direct Anti-tumor Activity --- p.41
Chapter 1.3.5.8 --- Modulating Activity in Endocrine System --- p.43
Chapter 1.3.5.9 --- Other Biological Activities --- p.43
Chapter 1.3.6 --- Possible Anti-cancer Mechanisms of Green Tea Epicatechins --- p.44
Chapter 1.3.6.1 --- Modulation of Anti-tumor Immunity --- p.44
Chapter 1.3.6.2 --- Direct Growth Inhibition by Controlling the Signal Transduction Pathways --- p.45
Chapter 1.3.6.3 --- Induction of Apoptosis and Cell Cycle Arrest --- p.46
Chapter 1.3.6.4 --- Inhibition of Tumor Metastasis --- p.47
Chapter 1.4 --- Aims and Scopes of This Investigation --- p.48
Chapter CHAPTER 2: --- MATERIALS AND METHODS
Chapter 2.1 --- Materials --- p.50
Chapter 2.1.1 --- Animals --- p.50
Chapter 2.1.2 --- Cell Lines --- p.50
Chapter 2.1.3 --- Sheep Red Blood Cells (SRBC) --- p.52
Chapter 2.1.4 --- "Cell Culture Medium, Buffers and Reagents" --- p.52
Chapter 2.1.5 --- Tea Extracts and Green Tea Epicatechins --- p.56
Chapter 2.1.6 --- Recombinant Cytokines --- p.57
Chapter 2.1.7 --- Vitamin Analogs --- p.59
Chapter 2.1.8 --- Taxol (Baccatin III N-benzoyl-β-phenyllisoserine ester) --- p.59
Chapter 2.1.9 --- 18β-Glycyrrhetinic Acid (18β-GA) --- p.60
Chapter 2.1.10 --- [methyl-3H] Thymidine (3H-TdR) --- p.60
Chapter 2.1.11 --- Liquid Scintillation Cocktail --- p.60
Chapter 2.1.12 --- Reagents and Buffers for Flow Cytometery --- p.61
Chapter 2.1.13 --- Reagents for DNA Extraction --- p.62
Chapter 2.1.14 --- Reagents for Total RNA Isolation --- p.63
Chapter 2.1.15 --- Reagents and Buffers for RT-PCR Study --- p.64
Chapter 2.1.16 --- Reagents and Buffers for Gel Electrophoresis --- p.67
Chapter 2.1.17 --- Reagents and Buffers for Western Blot Analysis --- p.68
Chapter 2.2 --- Methods --- p.77
Chapter 2.2.1 --- Culture of the Leukemic Cell Lines --- p.77
Chapter 2.2.2 --- "Isolation, Preparation and Culture of Primary Mouse Cells" --- p.77
Chapter 2.2.3 --- Determination of Cell Viability --- p.78
Chapter 2.2.4 --- [3H]-TdR Incorporation Assay --- p.79
Chapter 2.2.5 --- Cell Morphology Study --- p.79
Chapter 2.2.6 --- Apoptosis Study --- p.80
Chapter 2.2.7 --- Animal Studies --- p.81
Chapter 2.2.8 --- Gene Expression Study --- p.82
Chapter 2.2.9 --- Protein Expression Study --- p.85
Chapter 2.2.10 --- Statistical Analysis --- p.88
Chapter CHAPTER 3: --- THE ANTI-TUMOR ACTIVITIES OF TEA EXTRACTS AND PURIFIED GREEN TEA EPICATECHIN ISOMERS ON VARIOUS LEUKEMIC CELL LINES
Chapter 3.1 --- Introduction --- p.89
Chapter 3.2 --- Results --- p.91
Chapter 3.2.1 --- The Effects of Tea Extracts on Various Leukemia Cells --- p.91
Chapter 3.2.1.1 --- Differential Anti-proliferative Effect of Different Tea Extracts on Various Leukemic Cell Lines In Vitro --- p.91
Chapter 3.2.1.2 --- Differential Cytotoxic Effect of Different Tea Extracts on the Murine Lymphocytic Leukemia L1210 Cells In Vitro --- p.92
Chapter 3.2.1.3 --- Induction of Apoptosis in HL-60 Cells by Different Tea Extracts In Vitro --- p.92
Chapter 3.2.2 --- The Effects of Purified Green Tea Epicatechin Isomers on Various Leukemic Cell Lines --- p.101
Chapter 3.2.2.1 --- In Vitro Anti-proliferative Effect of Green Tea Epicatechin Isomers on Various Human and Murine Leukemic Cell Lines --- p.101
Chapter 3.2.2.2 --- In Vitro Cytotoxic Effect of Green Tea Epicatechin Isomers on Various Human and Murine Leukemic Cell Lines --- p.117
Chapter 3.2.2.3 --- Effects of Green Tea Epicatechin Isomers on the Differentiation of Myeloid Leukemia Cells --- p.131
Chapter 3.2.2.4 --- Apoptosis-Inducing Effect of Different Green Tea Epicatechin Isomers on HL-60 and JCS Cells --- p.134
Chapter 3.2.2.5 --- Effect of EGCG on the In Vivo Tumorigenicity of Leukemia JCS and L1210 Cells --- p.142
Chapter 3.3 --- Discussion --- p.144
Chapter CHAPTER 4: --- MECHANISTIC STUDIES ON THE ANTI PROLIFERATIVE AND APOPTOSIS-INDUCING ACTIVITIES OF GREEN TEA EPICATECHIN ISOMERS ON LEUKEMIA CELLS
Chapter 4.1 --- Introduction --- p.149
Chapter 4.2 --- Results --- p.152
Chapter 4.2.1 --- Combining Effect of EGCG and Physiological Differentiation Inducers on the Proliferation of HL-60 and JCS Cells --- p.152
Chapter 4.2.2 --- Combining Effect of EGCG and Cytokines on the Proliferation of JCS Cells --- p.155
Chapter 4.2.3 --- Combining Effect ofEGCG and Other Phytochemicals on the Proliferation of HL-60 and JCS Cells --- p.161
Chapter 4.2.4 --- Modulatory Effect of EGCG on the Expression of Apoptosis-regulatory Genes in HL-60 Cells --- p.168
Chapter 4.2.5 --- Modulatory Effect of EGCG on the Expression of Growth-related and Apoptosis-regulatory Proteins in HL-60 Cells --- p.170
Chapter 4.3 --- Discussion --- p.177
Chapter CHAPTER 5: --- EFFECTS OF GREEN TEA EPICATECHIN ISOMERS ON THE GROWTH AND DIFFERENTIATION OF MURINE HEMATOPOIETIC CELLS
Chapter 5.1 --- Introduction --- p.184
Chapter 5.2 --- Results --- p.186
Chapter 5.2.1 --- In Vitro Effects of EGCG on Murine Lymphocytes --- p.186
Chapter 5.2.1.1 --- In Vitro Effect of EGCG on the Proliferation of Murine Splenocytes --- p.186
Chapter 5.2.1.2 --- In Vitro Effect of EGCG on the Mitogen-induced Proliferation of Murine Splenocytes --- p.186
Chapter 5.2.1.3 --- Cytotoxic Effect of EGCG on Murine Lymphocytes --- p.189
Chapter 5.2.2 --- Primary Humoral Immune Response to SRBCin EGCG-treated Mice --- p.191
Chapter 5.2.3 --- In Vitro Studies of the Effects of EGCG on Murine Bone Marrow Cells --- p.192
Chapter 5.2.3.1 --- Effects of EGCG on the In Vitro Proliferation of Murine Bone Marrow Cells --- p.192
Chapter 5.2.3.2 --- The Combining Effect of EGCG and Growth Factors on the In Vitro Proliferation of Murine Bone Marrow Cells --- p.192
Chapter 5.2.3.3 --- In Vitro Cytotoxic Effect of EGCG on Murine Bone Marrow Cells --- p.196
Chapter 5.2.4 --- Effect of EGCG on the Differentiation of Murine Bone Marrow Cells --- p.199
Chapter 5.2.5 --- Combining Effects of EGCG and Growth Factors on the Morphology of Murine Bone Marrow Cells --- p.199
Chapter 5.3 --- Discussion --- p.204
Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.207
REFERENCES --- p.213
48
„Isolation of defense proteins from plant seeds and storage organs, and investigation on their potential applications“. 2012. http://library.cuhk.edu.hk/record=b5549533.
Annotation:
病原體感染是包括植物的高等生物的主要健康危害之一。為抵禦入侵者，大多數植物會製造防禦蛋白，包括凝集素、蛋白酶抑製劑、抗真菌蛋白、核糖核酸酶和核糖體失活蛋白，並分佈在不同的器官，如葉、根、種子和塊莖。一些植物防禦蛋白被發現能表現出多種生物活性，如抗腫瘤活性、抗細菌活性和抗病毒活性，能抵抗多種植物病原菌和人類病原體。因此，一些植物防禦蛋白可能有潛力用於治療人類疾病，或保護農作物免受感染。我們在研究中從不同的植物來源成功純化出各種防禦蛋白，包括：小芋頭塊莖中的血凝素、日本長芋中的凝集素、東北紅豆中的血凝素和抗真菌多肽、棕色芸豆中的凝集素、抗真菌多肽和胰蛋白酶抑製劑，玉豆一號中的凝集素以及小斑豆中的胰蛋白酶抑製劑。小芋頭血凝素被發現能誘導脾細胞的有絲分裂反應。日本長芋凝集素和東北紅豆血凝素被發現能對一些腫瘤細胞株（如乳腺癌MCF7細胞及鼻咽癌CNE2細胞）發揮抗增殖的作用。棕色芸豆凝集素能誘導脾臟細胞的有絲分裂反應以及抑制腫瘤細胞株（如乳腺癌MCF7細胞、肝癌HepG2及鼻咽癌CNE1和 CNE2細胞）的生長，而棕色芸豆抗真菌蛋白能抑制數種病原真菌物種的生長。研究這些防禦蛋白的生物活性有助找出其潛在應用價值，如藥用前景。
Infection from pathogens is one of the major health hazards in higher organisms including plants. To defend against harmful invaders, most plants produce a variety of defense proteins including lectins, protease inhibitors, antifungal proteins, ribonucleases and ribosome-inactivating proteins. They may be present in different organs of the plants, such as leaves, roots, seeds and tubers. Some of the plant defense proteins were found to exhibit a variety of biological activities such as anti-tumor activity, anti-bacterial activity and anti-viral activity that act against various plant pathogens and also some human pathogens. Therefore, some plant defense proteins may have potential for therapeutic applications in human diseases, or protecting the crops from infections.
This study involved purification of defense proteins from different plant sources. The proteins that were successfully isolated included a hemagglutinin from small taro tubers, a lectin from Japanese yam tubers, a lectin and an antifungal peptide from northeast red beans, a lectin, an antifungal peptide and a trypsin inhibitor from brown kidney beans, a lectin from French bean cultivar no. 1 and a trypsin inhibitor from mini pinto beans. The small taro hemagglutinin was found to induce mitogenic response in splenocytes. The Japanese yam lectin and northeast red bean hemagglutinin were found to exert anti-proliferative activity toward some tumor cell lines including MCF7 and CNE2 cells. The brown kidney bean lectin induced a mitogenic response from murine splenocytes as well as inhibited the growth of tumor cell lines including MCF7, HepG2, CNE1 and CNE2 cells, while the brown kidney bean antifungal protein inhibited the growth of several pathogenic fungal species including M. arachidicola, S. turcica and B. maydis. Studying the biological activities of these defense proteins helps to find out their potential applications like therapeutic uses.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Chan, Yau Sang.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2012.
Includes bibliographical references (leaves i-xvii).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
Abstract --- p.i-ii
論文摘要 --- p.iii
Acknowledgements --- p.iv
List of Publications --- p.v
Table of Contents --- p.vi-vii
List of Figures --- p.viii-ix
List of Tables --- p.x
List of Abbreviations --- p.xi
Chapter Chapter 1 --- Introduction on plant defense proteins
Chapter 1.1 --- General introduction to plant defense proteins --- p.1-2
Chapter 1.2 --- An overview on lectins --- p.3-18
Chapter 1.2.1 --- History of lectins --- p.3-6
Chapter 1.2.2 --- Classification of lectins --- p.7-11
Chapter 1.2.3 --- Biological activities of lectins --- p.12-16
Chapter 1.2.4 --- Applications of plant lectins --- p.16-18
Chapter 1.3 --- An overview on defensins --- p.18-25
Chapter 1.3.1 --- Types of defensins --- p.18-21
Chapter 1.3.2 --- Mechanism of anti-microbial activity of defensins --- p.22-23
Chapter 1.3.3 --- Application of defensins --- p.23-25
Chapter 1.4 --- An overview on trypsin inhibitors --- p.25-38
Chapter 1.4.1 --- Serpins --- p.26-28
Chapter 1.4.2 --- Kunitz-type protease inhibitors --- p.29-31
Chapter 1.4.3 --- Bowman-Birk protease inhibitors --- p.32-34
Chapter 1.4.4 --- Physiological functions of protease inhibitors --- p.35-38
Chapter 1.5 --- Aim of study --- p.38-41
Chapter Chapter 2 --- Isolation and characterization of a hemagglutinin from small taros and a lectin from yam tubers
Chapter 2.1 --- Introduction --- p.42-45
Chapter 2.2 --- Materials and Methods --- p.46-55
Chapter 2.3 --- Results --- p.56-78
Chapter 2.4 --- Discussion --- p.79-84
Chapter Chapter 3 --- Isolation and characterization of two defense proteins from seeds of Phaseolus vulgaris cv. “northeast red bean“
Chapter 3.1 --- Introduction --- p.85-86
Chapter 3.2 --- Materials and Methods --- p.87-93
Chapter 3.3 --- Results --- p.93-119
Chapter 3.4 --- Discussion --- p.120-129
Chapter Chapter 4 --- Isolation and characterization of three defense proteins from seeds of Phaseolus vulgaris cv. “brown kidney bean“
Chapter 4.1 --- Introduction --- p.130-131
Chapter 4.2 --- Materials and Methods --- p.131-136
Chapter 4.3 --- Results --- p.136-175
Chapter 4.4 --- Discussion --- p.176-189
Chapter Chapter 5 --- Isolation and characterization of a lectin from French bean cultivar no. 1 beans and a trypsin inhibitor from mini pinto beans
Chapter 5.1 --- Introduction --- p.190-191
Chapter 5.2 --- Materials and Methods --- p.191-194
Chapter 5.3 --- Results --- p.195-212
Chapter 5.4 --- Discussion --- p.213-221
Chapter Chapter 6 --- General discussion
Chapter 6.1 --- Summary on purification protocols of the defense proteins in the study --- p.222-228
Chapter 6.2 --- Chemical properties of the defense proteins in the study --- p.228-232
Chapter 6.3 --- Biological activities of the defense proteins in the study --- p.232-238
Chapter 6.4 --- Potential application of these defense proteins and future perspectives --- p.238-242
References --- p.i-xvi
49
„Persistence in the use of statins and the associated outcomes among Chinese patients with high risk for coronary heart disease“. 2004. http://library.cuhk.edu.hk/record=b5892118.
Annotation:
Cheng Wai Ring Caroline.Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.
Includes bibliographical references (leaves 74-84).
Abstracts in English and Chinese.
Acknowledgement --- p.i
Abstract --- p.ii
摘要 --- p.iv
Table of contents --- p.vi
Publications --- p.x
List of figures --- p.xi
List of tables --- p.xii
Abbreviations --- p.xiii
Chapter Chapter 1 --- Introduction
Chapter 1.1 --- Coronary Heart Disease --- p.2
Chapter 1.1.1 --- Epidemiology --- p.2
Chapter 1.2 --- Hypercholesterolemia and CHD --- p.3
Chapter 1.2.1 --- Atherosclerotic plaque and lipoprotein --- p.4
Chapter 1.2.2 --- NCEP ATP III guidelines --- p.5
Chapter 1.2.2.1 --- CHD risk assessment --- p.5
Chapter 1.2.2.2 --- Target lipid control --- p.8
Chapter 1.2.2.3 --- Therapeutic lifestyle changes --- p.9
Chapter 1.2.2.4 --- Pharmacological interventions --- p.10
Chapter 1.2.2.5 --- Adherence to lipid-lowering therapy --- p.13
Chapter 1.3 --- Adherence to drug therapy --- p.14
Chapter 1.3.1 --- Definition of adherence --- p.14
Chapter 1.3.2 --- Methods to assess adherence --- p.16
Chapter 1.3.2.1 --- Expressions of adherence measurements --- p.20
Chapter 1.3.3 --- Time effect on adherence --- p.21
Chapter 1.3.4 --- Predictors of adherence --- p.22
Chapter 1.3.5 --- Impact of poor adherence to statins --- p.23
Chapter 1.4 --- Objectives and hypotheses --- p.25
Chapter Chapter 2 --- Materials and Methods
Chapter 2.1 --- Study site --- p.27
Chapter 2.2 --- Patient selection criteria --- p.28
Chapter 2.2.1 --- Inclusion criteria --- p.28
Chapter 2.2.2 --- Exclusion criteria --- p.29
Chapter 2.3 --- Patient recruitment --- p.30
Chapter 2.4 --- Assessments --- p.32
Chapter 2.4.1 --- Adherence assessment --- p.32
Chapter 2.4.1.1 --- Electronic monitoring --- p.32
Chapter 2.4.1.2 --- Patient report --- p.33
Chapter 2.4.1.3 --- Pill count --- p.34
Chapter 2.4.1.4 --- Predictors of adherence --- p.34
Chapter 2.4.2 --- Clinical outcome assessment --- p.35
Chapter 2.4.2.1 --- Lipid control --- p.35
Chapter 2.4.3 --- Economic outcome assessment --- p.35
Chapter 2.4.3.1 --- Total direct medical cost --- p.35
Chapter 2.4.3.2 --- Healthcare cost per member per month --- p.36
Chapter 2.5 --- Sample size --- p.36
Chapter 2.6 --- Statistical analysis --- p.37
Chapter Chapter 3 --- Results
Chapter 3.1 --- Study sample --- p.40
Chapter 3.1.1 --- Demographic characteristics --- p.41
Chapter 3.1.2 --- Co-morbidity factors --- p.42
Chapter 3.2 --- Adherence measurement --- p.44
Chapter 3.2.1 --- Electronic monitoring --- p.44
Chapter 3.2.2 --- Patient report --- p.45
Chapter 3.2.3 --- Pill Count --- p.46
Chapter 3.2.4 --- Correlation among methods for measuring adherence --- p.47
Chapter 3.2.5 --- Trend of adherence and persistence over time --- p.48
Chapter 3.2.6 --- Independent predictors of adherence --- p.49
Chapter 3.3 --- Outcome assessment --- p.52
Chapter 3.3.1 --- Clinical outcomes --- p.52
Chapter 3.3.2 --- Economic outcomes --- p.52
Chapter 3.4 --- Association between adherence and clinical outcomes --- p.53
Chapter 3.4.1 --- Adherence and LDL-C reduction --- p.53
Chapter 3.4.2 --- Adherence and NCEP ATP III target --- p.55
Chapter 3.5 --- Association between adherence and economic outcomes --- p.55
Chapter 3.5.1 --- Adherence and healthcare utilization --- p.55
Chapter Chapter 4 --- Discussion and Conclusion
Chapter 4.1 --- Discussion --- p.59
Chapter 4.1.1 --- Accuracy of patient report and pill count --- p.59
Chapter 4.1.2 --- Persistence to statin therapy over time --- p.62
Chapter 4.1.3 --- Predictors for patient adherence --- p.63
Chapter 4.1.4 --- Clinical impacts of patient adherence --- p.66
Chapter 4.1.5 --- Economic impacts of patient adherence --- p.68
Chapter 4.1.6 --- Limitations --- p.70
Chapter 4.2 --- Conclusion --- p.71
References --- p.74
Appendices
Appendix A-1. Framingham risk scoring system for male --- p.86
Appendix A-2. Framingham risk scoring system for female --- p.87
Appendix B-1. Information sheet provided to nurses of the Cardiology clinic --- p.88
Appendix B-2. Information sheet provided to nurses of the Diabetes clinic --- p.89
Appendix B-3. Information sheet provided to nurses of the Lipid clinic --- p.90
Appendix C. Data collection form --- p.91
Appendix D. Instruction sheet provided to the study patient --- p.94
Appendix E. Unit cost of items from electronic dispensing record and Hong Kong Gazette 2003 for estimating total direct medical cost --- p.95
50
Wang, Xuanting. „Identification of SARS-CoV-2 Polymerase and Exonuclease Inhibitors and Novel Methods for Single-Color Fluorescent DNA Sequencing by Synthesis“. Thesis, 2021. https://doi.org/10.7916/d8-n6ah-nt76.
Annotation:
This dissertation is divided into two main sections describing major portions of my Ph.D. research: (1) development of two enzymatic assays for identifying inhibitors of SARS-CoV-2 RNA dependent RNA polymerase (RdRp) and the associated proofreading exonuclease complexes, two key enzymatic activities of SARS-CoV-2, the virus responsible for the COVID-19 pandemic and (2) the design and implementation of four novel single-color fluorescent DNA sequencing by synthesis (SBS) methods, including the synthesis of many of the key nucleotide analogues required for these studies.
In response to the COVID-19 pandemic, the first part of my research is focused on the discovery of potential therapeutics for combating coronavirus infections. Chapter 1 describes the identification of several polymerase and exonuclease inhibitors for SARS-CoV-2 using novel mass spectrometry-based molecular assays. SARS-CoV-2 has an exonuclease complex, which removes nucleotide inhibitors such as Remdesivir that are incorporated into the viral RNA during replication, reducing the efficacy of these drugs for treating COVID-19. Combinations of inhibitors of both the viral RdRp and the exonuclease could overcome this deficiency. Chapter 1 reports the identification of hepatitis C virus NS5A inhibitors Pibrentasvir and Ombitasvir as SARS-CoV-2 exonuclease inhibitors. In the presence of identified exonuclease inhibitors, RNAs terminated with the active forms of the prodrugs like Sofosbuvir, Remdesivir and Favipiravir were largely protected from excision by the exonuclease, while in the absence of exonuclease inhibitors, there was rapid excision. Viral cell culture studies also demonstrate significant synergy using this combination strategy. This study supports the use of combination drugs that inhibit both the SARS-CoV-2 polymerase and exonuclease for effective COVID-19 treatment.
Chapters 2-6 describe the single-color DNA SBS studies. Chapter 2 provides essential background on the structure of DNA, the DNA polymerase reaction, and several key DNA sequencing technologies, with an emphasis on the design of nucleotide analogues for the DNA SBS approach. Chapter 3 delineates a one-color fluorescent DNA SBS method based on a set of nucleotide reversible terminators (NRTs) comprising two orthogonal cleavable linkers, one fluorescent dye and one anchor. Chapter 4 describes a one-color hybrid DNA sequencing approach using a set of dideoxynucleotide analogues bearing two orthogonal cleavable linkers, one fluorophore and one anchor as well as a set of unlabeled NRTs. By introducing a pH responsive fluorophore into the design of nucleotide analogues, Chapter 5 demonstrates a novel type of single-color DNA SBS method using a set of NRTs comprising one pH-responsive fluorescent dye or one non-responsive fluorescent dye tethered with one cleavable linker. Chapter 6 presents another option for the single-color DNA sequencing technique using a set of deoxynucleotide analogues comprising the above pH responsive or non-responsive dyes tethered with a cleavable linker, along with a set of unlabeled NRTs. The one-color SBS approaches have the potential for higher sensitivity, miniaturization and cost effectiveness compared with four-color SBS methods.
Finally, Chapter 7 summarizes the SARS-CoV-2 antiviral drug discovery and one-color sequencing techniques and discusses potential follow-up research on these projects.