Regio- and stereoselectivity of oxidative coupling reactions of phenols : Spirodienones as construction units in lignin

Dimeric phenolic compounds lignans and dilignols form in the so-called oxidative coupling reaction of phenols. Enzymes such as peroxidases and lac-cases catalyze the reaction using hydrogen peroxide or oxygen respectively as oxidant generating phenoxy radicals which couple together according to certain rules. In this thesis, the effects of the structures of starting materials mono-lignols and the effects of reaction conditions such as pH and solvent system on this coupling mechanism and on its regio- and stereoselectivity have been studied. After the primary coupling of two phenoxy radicals a very reactive quinone me-thide intermediate is formed. This intermediate reacts quickly with a suitable nucleophile which can be, for example, an intramolecular hydroxyl group or another nucleophile such as water, methanol, or a phenolic compound in the reaction system. This reaction is catalyzed by acids. After the nucleophilic addi-tion to the quinone methide, other hydrolytic reactions, rearrangements, and elimination reactions occur leading finally to stable dimeric structures called lignans or dilignols. Similar reactions occur also in the so-called lignification process when monolignol (or dilignol) reacts with the growing lignin polymer. New kinds of structures have been observed in this thesis. The dimeric com-pounds with so-called spirodienone structure have been observed to form both in the dehydrodimerization of methyl sinapate and in the beta-1-type cross-coupling reaction of two different monolignols. This beta-1-type dilignol with a spirodienone structure was the first synthetized and published dilignol model compound, and at present, it has been observed to exist as a fundamental construction unit in lignins. The enantioselectivity of the oxidative coupling reaction was also studied for obtaining enantiopure lignans and dilignols. A rather good enantioselectivity was obtained in the oxidative coupling reaction of two monolignols with chiral auxiliary substituents using peroxidase/H2O2 as an oxidation system. This observation was published as one of the first enantioselective oxidative coupling reaction of phenols. Pure enantiomers of lignans were also obtained by using chiral cryogenic chromatography as a chiral resolution technique. This technique was shown to be an alternative route to prepare enantiopure lignans or lignin model compounds in a preparative scale.

Mikä on ihme ja mitä ihmeistä voidaan tietää? : Keskustelu ihmeistä uusimmassa englanninkielisessä uskonnonfilosofiassa

What is a miracle and what can we know about miracles? A discussion of miracles in anglophone philosophy of religion literature since the late 1960s. The aim of this study is to systematically describe and philosophically examine the anglophone discussion on the subject of miracles since the latter half of the 1960s. The study focuses on two salient questions: firstly, what I will term the conceptual-ontological question of the extent to which we can understand miracles and, secondly, the epistemological question of what we can know about miracles. My main purpose in this study is to examine the various viewpoints that have been submitted in relation to these questions, how they have been argued and on what presuppositions these arguments have been based. In conducting the study, the most salient dimension of the various discussions was found to relate to epistemological questions. In this regard, there was a notable confrontation between those scholars who accept miracles and those who are sceptical of them. On the conceptual-ontological side I recognised several different ways of expressing the concept of miracle . I systematised the discussion by demonstrating the philosophical boundaries between these various opinions. The first and main boundary was related to ontological knowledge. On one side of this boundary I placed the views which were based on realism and objectivism. The proponents of this view assumed that miraculousness is a real property of a miraculous event regardless of how we can perceive it. On the other side I put the views which tried to define miraculousness in terms of subjectivity, contextuality and epistemicity. Another essential boundary which shed light on the conceptual-ontological discussion was drawn in relation to two main views of nature. The realistic-particularistic view regards nature as a certain part of reality. The adherents of this presupposition postulate a supernatural sphere alongside nature. Alternatively, the nominalist-universalist view understands nature without this kind of division. Nature is understood as the entire and infinite universe; the whole of reality. Other, less important boundaries which shed light on the conceptual-ontological discussion were noted in relation to views regarding the laws of nature, for example. I recognised that the most important differences between the epistemological approaches were in the different views of justification, rationality, truth and science. The epistemological discussion was divided into two sides, distinguished by their differing assumptions in relation to the need for evidence. Adherents of the first (and noticeably smaller) group did not see any epistemological need to reach a universal and common opinion about miracles. I discovered that these kinds of views, which I called non-objectivist, had subjectivist and so-called collectivist views of justification and a contextualist view of rationality. The second (and larger) group was mainly interested in discerning the grounds upon which to establish an objective and conclusive common view in relation to the epistemology of miracles. I called this kind of discussion an objectivist discussion and this kind of approach an evidentialist approach. Most of the evidentialists tried to defend miracles and the others attempted to offer evidence against miracles. Amongst both sides, there were many different variations according to emphasis and assumption over how they saw the possibilities to prove their own view. The common characteristic in all forms of evidentialism was a commitment to an objectivist notion of rationality and a universalistic notion of justification. Most evidentialists put their confidence in science in one way or another. Only a couple of philosophers represented the most moderate version of evidentialism; they tried to remove themselves from the apparent controversy and contextualised the different opinions in order to make some critical comments on them. I called this kind of approach a contextualising form of evidentialism. In the final part of the epistemological chapter, I examined the discussion about the evidential value of miracles, but nothing substantially new was discovered concerning the epistemological views of the authors.

Understanding Human Drug Conjugating Enzymes; Regio- and Stereoselectivity in Sulfotransferase 1A3 and the UDP-Glucuronosyltransferases

Sulfotransferases (SULTs) and UDP-glucuronosyltransferases (UGTs) are important detoxification enzymes and they contribute to bioavailability and elimination of many drugs. SULT1A3 is an extrahepatic enzyme responsible for the sulfonation of dopamine, which is often used as its probe substrate. A new method for analyzing dopamine-3-O-sulfate and dopamine-4-O-sulfate by high-performance liquid chromatography was developed and the enzyme kinetic parameters for their formation were determined using purified recombinant human SULT1A3. The results show that SULT1A3 strongly favors the 3-hydroxy group of dopamine, which indicates that it may be the major enzyme responsible for the difference between the circulating levels of dopamine sulfates in human blood. All 19 known human UGTs were expressed as recombinant enzymes in baculovirus infected insect cells and their activities toward dopamine and estradiol were studied. UGT1A10 was identified as the only UGT capable of dopamine glucuronidation at a substantial level. The results were supported by studies with human intestinal and liver microsomes. The affinity was low indicating that UGT1A10 is not an important enzyme in dopamine metabolism in vivo. Despite the low affinity, dopamine is a potential new probe substrate for UGT1A10 due to its selectivity. Dopamine was used to study the importance of phenylalanines 90 and 93 in UGT1A10. The results revealed distinct effects that are dependent on differences in the size of the side chain and on the differences in their position within the protein. Examination of twelve mutants revealed lower activity in all of them. However, the enzyme kinetic studies of four mutants showed that their affinities were similar to that of UGT1A10 suggesting that F90 and F93 are not directly involved in dopamine binding in the active site. The glucuronidation of β-estradiol and epiestradiol (α-estradiol) was studied to elucidate how the orientation of the 17-OH group affects conjugation at the 3-OH or the 17-OH of either diastereomer. The results show that there are clear differences in the regio- and stereoselectivities of UGTs. The most active isoforms were UGT1A10 and UGT2B7 demonstrating opposite regioselectivity. The stereoselectivities of UGT2Bs were more complex than those of UGT1As. The amino acid sequences of the human UGTs 1A9 and 1A10 are 93% identical, yet there are large differences in their activity and substrate selectivity. Several mutants were constructed to identify the residues responsible for the activity differences. The results revealed that the residues between Leu86 and Tyr176 of UGT1A9 determine the differences between UGT1A9 and UGT1A10. Phe117 of UGT1A9 participated in 1-naphthol binding and the residues at positions 152 and 169 contributed to the higher glucuronidation rates of UGT1A10. In summary, the results emphasize that the substrate selectivities, including regio- and stereoselectivities, of UGTs are complex and they are controlled by many amino acids rather than one critical residue.