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

Esophageal squamous cell carcinoma (ESCC) is the predominant type of esophageal cancer in Asia. Cisplatin is commonly used as an agent for treating ESCC patients undergoing chemotherapy. However establishment of resistance over the course of treatment diminishes the clinical usefulness and is one reason explaining poor prognosis of ESCC patients. In order to gain insights into the mechanism of cisplatin resistance in ESCC, HPLC/nESI-MS/MS proteomic profiling was employed to examine the global protein alterations of cisplatin-resistant ESCC cell line HKESC2/CDDP comparing with its parental cisplatin-sensitive cell line HKESC2. Stable over-expression and knocked-down cell lines were established for pathway analysis and functional studies. Seventeen proteins were identified with more than 2-fold difference in expression levels. These proteins are involved in endoplasmic reticulum stress response, metabolic processes, DNA replication and repair, nucleotide binding and cell cycle control, while some of them are components of cytoskeletal proteins. Among them, 14-3-3σ was one of the most significantly upregulated proteins found in HKESC2/CDDP cells and its differential expression levels were validated using western blotting and real-time quantitative polymerase chain reaction. Pathway analysis revealed that ectopic overexpression of 14-3-3σ caused a general upregulation in DNA repairing genes. Furthermore, functional validation showed that elevated 14-3-3σ expression contributed considerably to the observed cisplatin resistance in HKESC2/CDDP cells. While knocking down 14-3-3σ expression reversed the above situations in SLMT1 cells. I conclude that up-regulation in 14-3-3σ, together with DNA repairing genes, contributes to the establishment of cisplatin resistance in HKESC2/CDDP cells. Knocking down 14-3-3σ expression sensitized ESCC cells to cisplatin treatment, and hence, opens a therapeutic opportunity for ESCC cisplatin resistance. / published_or_final_version / Surgery / Doctoral / Doctor of Philosophy

published_or_final_version / Nursing Studies / Master / Master of Nursing in Advanced Practice

HIV/AIDS remains one of the most pressing challenges facing South African society. South Africa has the highest number of people living with HIV as well as the highest number of people on HIV treatment globally, yet only 37% of persons eligible for treatment have access to treatment. The advent of HAART ushered in a new era in the treatment of HIV infection. HIV infection was no longer a life threatening terminal illness, HIV/AIDS became a chronic manageable disease. The full clinical benefit of HAART can only be achieved with near perfect adherence i.e. > 95%. This means taking the medication exactly as prescribed; on time, no missed doses, every day, lifelong. No other chronic medication requires such stringent adherence rates for optimal therapeutic benefit, which may mean the choice between life and death. Achieving near perfect adherence poses a serious challenge to health service providers and persons on treatment as typical adherence rates for medication prescribed over long periods are in the 50 – 75 % range. Persons on HAART live with the additional burden of drug resistance and limited treatment options if near perfect adherence rates are not achieved. The purpose of this qualitative study was to explore the factors that influence adherence to HAART. These factors may be related to the person, the health care team and system, the treatment regimen, the social and economic environment or to the effects of HIV disease. Factors may either negatively or positively influence a person’s ability to adhere optimally to their prescribed treatment. A small sample of thirteen participants were purposefully selected for this study. Data was collected using in-depth interviews which were tape recorded and transcribed for thematic analysis. The value of this study is that it may assist health care providers, persons on treatment and the health care system to better comprehend the challenges of lifelong optimal adherence to HAART. / Thesis (M.A.)-University of KwaZulu-Natal, Durban, 2011.

Skrandžio vėžys yra ketvirta pagal dažnį ir antra pagal mirtingumą onkologinė liga pasaulyje. Sergančių šia liga 5 metų išgyvenamumas siekia tik 25 proc. Esant didelei ligos atkryčio rizikai dažniausiai skiriama adjuvantinė chemoterapija, nors daugumoje atsitiktinių imčių studijų statistiškai reikšmingo išgyvenamumo pagerėjimo nenustatyta. Dažniausia skrandžio vėžio gydymo nesėkmės priežastis yra lokoregioninis recidyvas (40-65 proc. ligonių, kuriems atliktos radikalios operacijos) ir pilvaplėvės metastazės. Siekiant sumažinti lokoregioninių recidyvų dažnį, pradėtas taikyti suderintas chemospindulinis gydymas. Šio tyrimo tikslas- nustatyti adjuvantinio gydymo efektyvumą po radikalių skrandžio vėžio operacijų su D2 limfadenektomija ir pagrįsti šio gydymo metodo tikslingumą. Darbo tikslui įgyvendinti buvo suformuluoti šie uždaviniai. 1. Įvertinti radikaliai dėl skrandžio vėžio operuotų su D2 limfadenektomija pacientų išgyvenamumą taikant adjuvantinį chemospindulinį gydymą arba adjuvantinę chemoterapiją. 2. Įvertinti adjuvantinio chemospindulinio gydymo toksiškumą po radikalių operacijų su D2 limfadenektomija, palyginti jį adjuvantinės chemoterapijos 5-fluoruracilu ir leukovorinu sukeliamu toksiškumu. 3. Įvertinti ankstyvos pooperacinės intraperitoninės chemoterapijos toksiškumą ir palyginti jos efektyvumą taikant su adjuvantiniu chemospinduliniu gydymu. 4. Palyginti gyvenimo kokybę po radikalių operacijų dėl skrandžio vėžio atliekant gastrektomiją ir subtotalinę skrandžio... [toliau žr. visą tekstą] / Gastric cancer is the fifth most common cancer and the second leading cause of cancer-related death worldwide. The 5-year survival rate of these patients is approx. 25. Adjuvant chemotherapy is frequently used for treatment, despite the fact that many randomized studies failed to demonstrate a better patient survival. The high rate of recurrence, even in patients undergoing state-of-the art curative resection, suggests that effective adjuvant chemoradiation and chemotherapy might indeed be an attractive concept to improve the overall outcomes of patients with gastric cancer. The aim of this study was to evaluate the effectiveness of the adjuvant therapy after curative resection with D2 lymphadenectomy for gastric cancer and determine its role in the treatment of cancer patients. The goals of this study were: 1. To compare the survival of patients receiving adjuvant chemoradiation or adjuvant chemotherapy after the curative resection with D2 lymphadenectomy for gastric cancer. 2. To evaluate the toxicity of the adjuvant chemoradiation after the curative resection with D2 lymphadenectomy for gastric cancer and to compare it with the toxicity caused by adjuvant chemotherapy with 5- Fluorouracil and Leucovorin. 3. To assess the toxicity of the early postoperative intraperitoneal chemotherapy and compare its effectiveness with the combined intraperitoneal chemotherapy and adjuvant chemoradiation therapy. 4. To compare the quality of life after the total and subtotal... [to full text]

Two experiments were conducted to evaluate the effects of arginine, vitamin E, and their combination on cardiopulmonary performance and nitric oxide synthase (NOS) activity in broilers undergoing cold-induced ascites (pulmonary hypertension) after an acute challenge with epinephrine. One day-old male broilers ( n = 25/treatment) were supplemented with arginine (0.3% water supplementation), vitamin E (400 IU/L or kg, water or feed supplementation; experiment 1 and 2 respectively) or both compounds, above NRC (1994) recommendations. From day 28 to 42, a catheterization was performed (n = 8/treatment) to evaluate pulmonary and mean arterial pressures, and heart rate. After recording baseline values, two injections of epinephrine (1 or 0.5 mg/kg body weight, experiment 1 and 2, respectively) were administered in a 20 min interval. The NOS activity was estimated through the conversion of 14C-arginine to 14C-citrulline in isolated pulmonary arteries ( n = 8/treatment). Overall, the time taken for pulmonary arterial pressure to return to values no different from initial baseline levels was longer for the birds consuming high levels of vitamin E (alone or in combination with arginine) compared to birds supplemented only with arginine. Although NOS activity was highly variable, birds fed with high arginine levels tended to have the lowest NOS activity. These results showed that vitamin E supplementation at the levels used in these experiments has detrimental effects on cardiopulmonary performance, and does not improve NOS activity in isolated pulmonary arteries.

Orientador: Jacqueline Pontes Monteiro / Banca: Daniela Saes Sartorelli / Banca: Carlos Alberto Scrideli / Banca: Maria Rita Marques de Oliveira / Banca: Maria Jacira Silva Simães / Resumo: As crianças com câncer necessitam manter o crescimento linear e o ganho de peso para uma melhor evolução clínica. Assim, o tratamento da criança com câncer deve garantir não só sua sobrevivência, mas proporcionar crescimento e desenvolvimento adequados. A avaliação nutricional é utilizada para reconhecer o paciente com maior facilidade de desenvolver a má nutrição garantindo assim um planejamento nutricional adequado. O objetivo do presente estudo foi avaliar o estado nutricional de crianças antes, durante e após o tratamento quimioterápico. Foram avaliadas 14 crianças com câncer, com idade entre 5 a 15 anos, divididas em dois grupos: Grupo Tumores Sólidos e Grupo Tumores Hematológicos. Estas foram avaliadas através dos dados de ingestão alimentar, antropometria, análise por Impedância Bioelétrica (BIA) e por Diluição Isotópica. Os resultados encontrados demonstram que os pacientes apresentaram um aumento de peso, estatura, índice de massa corporal, circunferência do quadril, da cintura e do braço, da prega cutânea subescapular, assim como o aumento da massa gorda pela técnica de diluição isotópica durante o tratamento quimioterápico, no Grupo Tumores Hematológicos. No Grupo Tumores Sólidos as crianças apresentaram uma diminuição da massa magra, quando analisado pela impedância bioelétrica. Houve uma correlação positiva entre a prega cutânea tricipital e a massa gorda obtida através da BIA e pela Diluição Isotópica. A circunferência muscular do braço também se correlacionou com a massa magra estimada pela BIA e pela Diluição Isotópica. Não houve mudança na ingestão energética e de macronutrientes nos três momentos da pesquisa. Esses resultados demonstram que os pacientes com tumores hematológicos apresentaram aumento de peso, estatura e de massa gorda, fato este não ocorrido no Grupo Tumores Sólidos. / Abstract: Children with cancer need to maintain growth and weight gain to a better clinic evolution. So, the treatment of children with cancer must not only guarantee the survival but also adequate growth and development. The nutritional assessment is useful to recognize the patient who is more accessible to develop malnutrition leading to an adequate nutritional plan. The aim of this study was to assess the nutritional status in children before, during and after chemotherapy treatment. In this study, 14 children with cancer were evaluated ranging from 5-15 years old devided into two groups: Hematologic Tumor Group and Solid Tumor Group. They were evaluated through dietary intake, anthropometry, bioimpedance analysis (BIA) and isotopic dilution. The results show that the patients increase weight, stature, body mass index, hip, waist and arm circumferences, subscapular skinfold thickness, as well as an increase in fat-mass with isotopic dilution technique during the chemotherapy treatment, in the Hematologic Tumors Group. In the Solid Tumor Group the children showed a reduction in fat free mass when assessed by bioelectric impedance. There was a positive correlation between triceps skinfold thickness and fat-mass with BIA and Isotopic Dilution technique. The arm muscle circumference correlated with the fatfree- mass estimated by the BIA and Isotopic Dilution. There was no change in energetic intake and macronutrients at the three moments of the research. These results demonstrate that patients with hematologic tumors had an increase in body weight, stature and fat-mass, which was not observed in Solid Tumors Group. The good correlation between anthropometry and isotopic dilution technique and BIA show its applicability in clinical practice. / Doutor

This project has focused on exploring the application of Baylis-Hillman (BH) {a.k.a. Morita-Baylis-Hillman (MBH)} scaffolds in the construction of various compounds with medicinal potential. A series of 2-nitrobenzaldehydes has been treated under BH conditions, with two different activated alkenes, viz., (MVK) and methyl acrylate, using (DABCO) or (3-HQ) as catalyst. While most of the BH reactions were carried out at room temperature, some reactions were conducted using microwave irradiation. The resulting BH adducts have been subjected to dehydration, conjugate addition and allylic substitution to obtain appropriate intermediates, which have been used in turn, to synthesize possible lead compounds, viz., cinnamate esters as HIV-1 integrase inhibitors, 3-(aminomethyl)quinolines and quinolones as anti-malarials and cinnamate ester-AZT conjugates as dual-action HIV-1 integrase-reverse transcriptase (IN-RT) inhibitors. Conjugate addition reactions of methyl acrylate-derived BH β-hydroxy esters with the amines, piperidine, propargylamine and 2-amino-5-(diethylamino)pentane, has afforded a range of products as diastereomeric mixtures in moderate to excellent yields. Catalytic hydrogenation of the aminomethy β-hydroxy esters derivatives, using a palladium-oncarbon (Pd-C) catalyst, has afforded the corresponding, novel 3-aminomethyl-2- quinolone derivatives in moderate yields. Effective allylic substitution reactions of the MVK-derived BH β-hydroxy ketones (via a conjugate addition-elimination pathway) using in situ-generated HCl has afforded the corresponding α-chloromethyl derivatives, which have been reacted with various amines, including piperidine, piperazine, propargylamine and 2-amino-5-(diethylamino)pentane, to yield α-aminomethyl derivatives. Catalytic hydrogenation of selected α-aminomethyl derivatives, using a Pd-C catalyst, has afforded the corresponding, novel 3- (aminomethyl)-2-methylquinoline derivatives in low to moderate yields. A bioassay, conducted on a 6-hydroxy-2-methyl-3-[(piperidin-1-yl)methyl]quinoline isolated early in the study indicated anti-malarial activity and prompted further efforts in the synthesis of analogous compounds. Reaction of the methyl acrylate-derived BH adducts with POCl3 has provided access to α-(chloromethyl)cinnamate ester derivatives, which have been aminated to afford α- (aminomethyl)cinnamate ester derivatives as potential HIV-1 integrase inhibitors. The α- (propargylaminomethyl)cinnamates were used, in turn, as substrates for the “click chemistry” reaction with 3'-azido-3'-deoxythymidine (AZT– an azide and an established reverse transcriptase HIV-1 inhibitor) to afford cinnamate ester-AZT conjugates as potential dual-action HIV-1 integrase-reverse transcriptase (IN-RT) inhibitors. Computer modelling and docking studies of a cinnamate ester-AZT conjugate into the HIV-1 integrase and reverse transcriptase active-sites revealed potential hydrogen-bonding interactions with amino acid residues within the receptor cavities. The isolated products have been appropriately characterized using IR, 1- and 2-D NMR and HRMS techniques, while elucidation of the stereochemistry of the double bond in the BH-derived halomethyl derivatives has been assigned on the basis of NOE, computer modelling and X-ray crystallographic data.

Published in Supportive Care in Cancer 2014;22(8):2161-6. DOI:10.1007/s00520-014-2182-7 / 京都大学 / 0048 / 新制・課程博士 / 博士(社会健康医学) / 甲第18548号 / 社医博第59号 / 新制||社医||8(附属図書館) / 31448 / 京都大学大学院医学研究科社会健康医学系専攻 / (主査)教授 武藤 学, 教授 佐藤 俊哉, 教授 千葉 勉 / 学位規則第4条第1項該当 / Doctor of Public Health / Kyoto University / DFAM

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