Poon, Ching Ho. / "October 2010." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 95-102). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.3 / TABLE OF CONTENTS --- p.4 / ABSTRACT --- p.6 / 摘要 --- p.8 / ABBREVIATIONS --- p.10 / LIST OF FIGURES --- p.11 / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.14 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.26 / Chemicals and materials --- p.26 / Mammalian cell culture --- p.27 / Reporter gene assay --- p.28 / Semi-quantitative real-time PCR --- p.30 / RNA degradation assay --- p.31 / Western Blotting --- p.32 / Electrophoretic mobility shift assay (EMSA) --- p.34 / EROD assay in intact cells --- p.35 / Statistical Analysis --- p.35 / Chapter CHAPTER 3 --- Two citrus bioflavonoids suppress DMBA-induced CYP1 gene expression and activity via antagonizing AhR transactivation activity --- p.36 / Chapter CHAPTER 4 --- Modulation of AhR-mediated CYP1 enzymes transcription by licorice flavonoids isoliquiritigenin --- p.60 / Chapter CHAPTER 5 --- Protein kinase signaling pathways are not involved in the modulation of AhR transactivation activity by flavonoids --- p.81 / Chapter CHAPTER 6 --- SUMMARY --- p.94 / BIBLIOGRAPHY --- p.97

Microcosm studies were conducted beginning with three xylene isomers: ortho-xylene, meta-xylene and para-xylene; and continued with the four mononitroxylene (MNX) isomers, culminating with testing ten dinitroxylene (DNX) isomers. Soil samples were obtained from a historically contaminated site with high levels of dinitrotoluene (DNT), trinitrotoluene (TNT) and dinitroxylene (DNX) and used as the inoculum for microcosm tests. The microcosm method of different isomers was based on the previous work on biodegradation of nitrotoluene. As it was demonstrated previously that 2,4-DNT degrading bacteria were present at the site, it was hypothesized that these may be capable of transforming or cometabolizing some of DNX isomers. Thus, DNX cometabolism studies were conducted in the presence of 2,4-DNT degrading bacteria. The presence of xylene and 2,4-DNT degrading was confirmed in this thesis. Meanwhile, several MNX and DNX isomers showed degradability in microcosm studies. Cometabolism studies showed that four DNX isomers could be cometabolized by 2,4-DNT enrichment.

Not available / text

Thesis (Ph.D.)--University of Tulsa, 1995. / Includes bibliographical references (leaves 197-214).

Thesis (Ph. D.)--Oregon State University, 2010. / Printout. Includes bibliographical references (leaves 104-108). Also available on the World Wide Web.

Most pharmaceutically relevant proteins and many extracellular proteins contain disulfide bonds. Formation of the correct disulfide bonds is essential for stability in almost all cases. Disulfide containing proteins can be rapidly and inexpensively overexpressed in bacteria. However, the overexpressed proteins usually form aggregates inside the bacteria, called inclusion bodies, which contains inactive and non-native protein. To obtain native protein, inclusion bodies need to be isolated and resolubilized, and then the resulting protein refolded in vitro. In vitro protein folding is aided by the addition of a redox buffer, which is composed of a small molecule disulfide and/or a small molecule thiol. The most commonly used redox buffer contains reduced and oxidized glutathione. Recently, aliphatic dithiols and aromatic monothiols have been employed as redox buffers. Aliphatic dithiols improved the yield of native protein as compared to the aliphatic thiol, glutathione. Dithiols mimic the in vivo protein folding catalyst, protein disulfide isomerase, which has two thiols per active site. Furthermore, aromatic monothiols increased the folding rate and yield of lysozyme and RNase A relative to glutathione. By combining the beneficial properties of aliphatic dithiols and aromatic monothiols, aromatic dithiols were designed and were expected to increase in vitro protein folding rates and yields. Aromatic monothiols (1-4) and their corresponding disulfides (5-8), two series of ortho- and para-substituted ethylene glycol dithiols (9-15), and a series of aromatic quaternary ammonium salt dithiols (16-17) were synthesized on a multigram scale. Monothiols and disulfides (1-8) were utilized to fold lysozyme and bovine pancreatic trypsin inhibitor. Dithiols (11-17) were tested for their ability to fold lysozyme. At pH 7.0 and pH 8.0, and high protein concentration (1 mg/mL), aromatic dithiols (16, 17) and a monothiol (3) significantly enhanced the in vitro folding rate and yield of lysozyme relative to the aliphatic thiol, glutathione. Additionally, aromatic dithiols (16, 17) significantly enhance the folding yield as compared to the corresponding aromatic monothiol (3). Thus, the folding rate and yield enhancements achieved in in vitro protein folding at high protein concentration will decrease the volume of renaturation solution required for large scale processes and consequently reduce processing time and cost.

The fluorescence spectrum of many polycyclic aromatic compounds (PACs) depends upon solvent polarity. The emission spectrum of PAC monomers consists of several major vibronic bands labeled I, II, etc., in progressive order. Emission intensity enhancement of select bands is observed in polar solvents.

In 1933, benzo[a]pyrene, a hydrocarbon, which was and still is of very great importance for cancer research, was isolated from coal tar and also was synthesized. In recent years more than 450 synthetic compounds have been found to be carcinogenic, and more than 200 are polycyclic aromatic hydrocarbons, their derivatives and analogues. Recently a new polynuclear hydrocarbon, a seven ring compound naphtho[2,1-a]perylene was synthesized in This Laboratory Physiological tests have revealed this compound to be a potent carcinogen. This experience has prompted a new initiative to prepare a number of related compounds of this type in order to bring further insight to the relation between chemical structure and the mechanism of physiological activity. During the synthesis of the four new seven fused aromatic ring systems and a new eight fused aromatic ring system, several modifications and improvement of existing synthetic procedures were made. A recently published modification of the Rosenmund-von Braun method of nitrile synthesis was successfully applied to the preparation of 2-(2-naphthylmethyl)benzonitrile. It was found that 2-(2-naphthylmethyl)phenyl-1-naphthyl ketone and 2-(2-naphthylmethyl)phenyl-2-naphthyl ketone could be prepared by the reaction of a Grignard reagent with a nitrile as well as by the inverse addition of a Grignard reagent to the appropriate acid chlorides. The alumina cyclodehydrogenation procedure was confirmed to be the, only method of synthesis that yields 12-(1-naphthyl)-benz[a]anthracene from its precursor ketone. The yield of 12-(2-naphthyl)benz[a]anthracene was increased from 61% to 83% when anhydrous hydrogen fluoride was used in place of 48% hydrogen bromide and glacial acidic acid as the cyclodehydration media of the precursor ketone. A new cyclodehydrogenation procedure was developed. This procedure, which employs a mixture of aluminum chloride-stannic chloride and alumina, was used to prepare a new hydrocarbon, naphtho[l,2-a]-perylene. An aluminum chloride-sodium chloride melt permeated with carbon dioxide was successfully employed in the preparation of naphtho[2,2-1]benzo- (a]pyrene, naphtho(l,2-l]benzo[a]pyrene, and naphtho- (2,3-1]benzo[a]pyrene. It was shown that high temperature gas chromatography with ionization detectors can be used with success to analyze all of the above discussed ketones, benz[a]anthracenes as well as the new perylene and pyrenes. Additional support of the validity of the. structures of naphtho[1,2-a]perylene and naphtho- [2,1-1]benzo[a]pyrene was provided when the cyclodehydrogenation of these hydrocarbons yielded one and the same product, naphtho[l,7,8-efg]anthanthrene. It was observed that the correlation between color and structure of the newly prepared hydrocarbons follows the principles of annelation. When the ultraviolet and visible spectra peak frequencies were compared it was found that the values and positions of the peaks follow the principles of the annelation method. The examination of the infrared absorption spectra revealed that naphtho[l,2-a]perylene, naphtho[2,1l-1]benzo[a]pyrene, naphtho[1,2-1]benzo[a]-pyrene, and naphtho[2,3-1]benzo[a]pyrene exhibited Peaks at all four, "solo," "duo," "trio," and "quartet", carbon-hydrogen vibration regions, but as expected naphtho[1,7,8-efg]anthanthrene had the "quartet" carbon-hydrogen peak missing between 770 and 755 cm. which further substantiated the validity of the naphtho[1,7,8-efg]anthanthrene structure. The TNF molecular adducts of the five newly prepared compounds, naphtho[1,2-a]perylene, naphtho- [2,1-1]benzo[a]pyrene, naphtho1l,2-1]benzo[a]pyrene, naphtho[2,3-1]benzo[a]pyrene, and naphtho[1,7,8-efg]-anthanthrene were prepared and their melting points recorded. In order to ascertain the structures of the naphtho[1,2-a]perylene and the naphtho[1,2-l]benzo[a]-pyrene obtained from the cyclodehydrogenation of 12-(1-naphthyl)benz[a]anthracene and 12-(2-naphthyl)-benz[a]anthracene, respectively, other routes of synthesis were undertaken. The hydrocarbon, 11-(1-naphthyl)benz[a]anthracene, was prepared by the reaction of 1-naphthylmagnesium bromide with 11-keto-5,6,8,9,10,11-hexahydrobenz[a]- anthracene which on distillation under reduced pressure gave 11-(1-naphthyl)-5,6,8,9-tetrahydrobenz[a]anthracene and on aromatization yielded 11-(1-naphthyl)benz[a]-anthracene. When 1-naphthyl magnesium bromide was allowed to react with the 11-keto-8,9,10,11-tetrahydrobenz[a]anthracene, 11-(1-naphthyl)-8,9~dihydrobenz[a]anthracene was obtained when distilled under reduced pressure. This, likewise, gave 11-(1-naphthyl)- benz[a]anthracene on aromatization. Naphtho[1,2-a]perylene was synthesized unequivocally from 11-(1-naphthyl)benz[a]anthracene via a cyclodehydrogenation reaction. The hydrocarbon, 1-(1-naphthyl)-1,2,3,4-tetrahydrobenz[a]anthracene, was prepared by the reaction of 1-naphthylmagnesium bromide with 1-keto-1,2,3,4- tetrahydrobenz[a]anthracene. On distillation under reduced pressure 1-(1-naphthyl)-1,2,3,4-tetrahydrobenz[a]anthracene was obtained. Under the conditions of an aromatization procedure, naphtho[1,2-1]benzo[a]-pyrene was obtained. The hydrocarbons 11-(1l-naphthyl)-5,6,8,9-tetrahydrobenz[a]anthracene, 11-(1-naphthyl)-8,9-dihydrobenz[a]anthracene, 11-(1-naphthyl)benz[a]-anthracene, 1-(1-naphthyl)-1,2,3,4-tetrahydrobenz[a]anthracene are additional new compounds. / Ph. D.

The process of C. K. Bradsher for preparing mesosubstituted anthracene and 1,2-benzanthracene derivatives has been widely extended. The purpose of this research was to extend this process to the thiophene series, in addition to continuing some work on the 9-(dimethyl)- phenylanthracene series and some newly developed l,2-dimethyl-10-phenyl)-anthracene series. 2'-Thienyl-2-benzylbenzophenone was prepared and cyclized to 9-(2-thienyl)-anthracene by the conventional methods in good yields. 3'-Thienyl-2-benzylbenzophenone was prepared in a different way, indirect and tedious. Thiophene was iodinated to tetraiodothiophene and tetraiodothiophene was reduced with aluminum-amalgam to 3-iodothiophene. 3-Thienylmagnesiumiodide was prepared by the entrainment technique and condensed with 2-cyanodiphenylmethane to 3'-thienyl-2-benzylbenzophenone. This ketone was cyclized to 9-(3'-thienyl)-anthracene by the standard acid mixture. 2-(1-Naphthylmethyl)-phenyl-2-thienylketimine formed from the condensation between 2-(1'-naphthylmethyl)- benzonitrile and 2-thienylmagnesiumbromide resisted hydrolysis to the corresponding ketone but cyclized quantitatively to 10-(2-thienyl)-l,2-benzanthracene. Following the classical Bradsher process, the reaction between 2-thienylmagnesiumbromide and o-chlorobenzaldehyde was unsuccessful because the hydrol formed was so reactive that it polymerized rapidly and ended up with tars. A cross-condensation reaction between 2-ethynylmagnesium bromide and o-chlorobenzylchloride was utilized to give the expected 2-(2'-chlorobenzyl)- thiophene in one step. This new procedure has been widely extended to the other series. It is preferred to. the conventional Bradsher process and the Friedel-Crafts method in terms of time, chemicals, and unequivocal products formed. 2-(2'-Chlorobenzyl)-thiophene so formed by the new procedure was converted by von Braun reaction to the corresponding nitrile and the nitrile was allowed to react with phenylmagnesiumbromide to give 2-(2-thienylmethyl)-benzophenone. The above ketone was cyclized almost quantitatively to 4-phenyl-thiophanthrene. The thiophene-containing ketones prepared in this research are very viscous oils. In order to have crystalline derivatives for identification purposes these ketones were oxidized to the corresponding diketones. 2-Thienyl-2-benzylbenzophenone and 2-(2'-thienylmethyl)- benzophenone gave the identical diketone, 2-benzoyl-2'- thienyl-benzophenone. 3 1 -Thienyl-2-benzylbenzophenone was oxidized to anthraquinone by using 25% sulfuric .acid, acetic acid and sodium dichromate. The formation of anthraquinone indicates that the monoketone cyclized first and then split off the thiophene ring. Apparently, the 3-thienyl group is more susceptible to oxidation than the 2’-thienyl group. A study of the oxidation of anthracene derivatives for structure proof was made using 9-phenylanthracene as a model compound. It was oxidized to 10-phenyl-10-hydroxyanthrone which resisted further oxidation to anthraquinone. It is obvious that the tertiary alcohol system stabilized the phenyl ring. In continuing the work on the 9-(dimethyl)-phenylanthracene series, the yields of three ketimine salts and one ketone were improved. Satisfactory analytical data was obtained for 2’,6’-dimethyl-2-benzylbenzophenone. In an attempt to overcome the steric effects offered by 2,6-ortho groups, 2-(2',3'-dimethylbenzyl)-benzonitrile was prepared. The starting material for this series, l-bromo-2,3-dimethylbenzene has been prepared in 47% yield by diazotization in comparison with the previous yield of 30%. / Doctor of Philosophy

LD2668 .T4 1966 T458 / Master of Science