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In this article, I argue that the ritual experience of water-baptism plays an essential role in Paul's metaphorical expression and rhetorical purpose in 1 Corinthians 12:13. To explore the role of baptism, I use conceptual blending theory from cognitive linguistics to define and demonstrate the metaphorical ways in which ritual functions in the human mind. In so doing, I emphasize the performance of a ritual itself and the contextual perception of its performance, arguing for a metaphorical relationship between the two. I apply conceptual blending analysis to interpret the complex interplay of three metaphors in 1 Corinthians 12:13. I argue that Paul forms a conceptual blend of three metaphors in this verse, and that baptism, the water-rite, plays a pivotal role in this blend by providing the physical pattern of immersion and the cultural understanding of this immersion as a new belonging. Using baptism, Paul achieves his purpose of re-picturing the reception of the Spirit and appealing for social union. This verse thus presents an excellent case of the role of ritual in the emergence of early Christianity and the explanatory power of ritual studies to the New Testament texts.

Instead of blurring the oral and the literary media in antiquity (R. Bultmann and B. Gerhardsson) or dividing them with unsatisfying principles (J.D.G. Dunn), this article follows recent scholarship on orality to explore the mechanical operation of ancient scribal memory as the oral-written interface. In so doing, I argue that the agreement of order between the Synoptic Gospels is characteristic of memory-based utilizations of written texts and does not necessarily indicate the scribes’ visual contact with those texts. It is, rather, the very high degree of verbal agreement that indicates Matthew’s frequent visual contact with Q 10–11 and 12–13 throughout the gospel, even when following Mark’s narrative sequence by memory. This approach explains the infrequent micro-conflations on the Two Document Hypothesis (2DH) with a more mechanically probable procedure, and so strengthens the argument that the 2DH is more feasible than the Two Gospel Hypothesis and the Farrer-Goulder Hypothesis.

The amino acid-containing lithocholic acids (LCA) represent a new class of human sialyltransferase (ST) inhibitors. In this study, we have reported their design, synthesis, and inhibitory activity against human STs. Among these derivatives, D-Glu-LCA 7, L-Asp-L-Asp-LCA 13, and L-Asp-L-Asp-Gly-Gly-LCA 22 with specific amino acid sequence were the most active ones with IC50 values of 2.3–5.6 and 4.2-6.2 μM toward α-2,3-ST and α-2,6-ST, respectively. The current study demonstrates that the new class of ST in- hibitors inhibit cell migration in breast cancer cells by preventing closure of the wound rather than involv- ing a direct antiproliferative effect.

Based on the optimum deposition conditions of ZrN thin film from our previous study, by varying oxygen flow rate ranging from 0 to 8 sccm, nanocrystalline ZrN x O y thin films were deposited on p-type (100) Si substrates using hollow cathode discharge ion-plating (HCD-IP) system. The objective of this study was to investigate the effect of oxygen content on the composition, structure and properties of the ZrN x O y thin films. The oxygen content of the thin film, determined using X-ray photoelectron spectroscopy (XPS), increased with increasing oxygen flow rate. As the oxygen content increased, the color of the ZrN x O y thin film changed from golden yellow to blue and then slate blue, and the microstructure observed by scanning electron microscopy (SEM) varied from columnar structure to finer grains and finally flat and featureless structure. Phase separation of ZrN x O y to ZrN and monoclinic ZrO 2 was found from X-ray diffraction (XRD) patterns when the oxygen content was higher than 9.7 at.%. The hardness of the film slightly increased as the oxygen content was less than 9.7% and then decreased to 15.7 GPa, a typical hardness of ZrO 2 phase, as the oxygen content further increased. The total residual stress of the film was measured using an optical method, and the residual stresses of ZrN and ZrO 2 phases were determined separately using modified XRD sin 2 ψ method. The total stress was close to the stress in ZrN phase as the ZrO 2 fraction was less than 30%, and was close to that in ZrO 2 phase as the ZrO 2 fraction was over 30%. The electrical resistivity of the film increased significantly with the increase of oxygen content. The film properties showed consistent trend with phase separation. As the fraction of ZrO 2 phase was small, the apparent properties of the films were more close to those in ZrN. When ZrO 2 fraction was over 30%, the films mainly exhibited the properties of ZrO 2 .

The first series of 5'-triazole cytidines la-d and adenosines 2a-c was prepared and evaluated for inhibitory potency toward a-2,3-sialyltransferase. The synthesis of target compounds was achieved by converting the 5'-alcohol of protected nucleosides to the azide derivatives, which were then coupled with the alkynes by copper(I)-catalyzed cycloaddition to give protected 5'-triazole nucleosides 3a-d/7a-c, followed by deprotection. 5'-Triazole adenosines 2a-c were less efficient inhibitors of α-2,3-sialyltrans-ferase than their cytidine analogues la-d. 1d was the most active compound with an IC 50 of 37.5 uM. These results suggested that the hydrophobic functionality and the cytidine group are clearly required for the improved binding.

A new approach to the synthesis of anomeric sulfur analogues of CMP-Neu5Ac containing alkane or arene linkage la-d is described. The procedure involves the high β-stereoselectivity in sialylation of the peracetylated sialic acid methyl ester 4 with mercaptoalkyl (aryl) trichloroacetate 5a-d, followed by selective deprotection of the trichloroacetyl group to the corresponding hydroxyalkyl and hydroxyaryl thioglycosides 2a-d. Subsequent O-phosphitylation of 2a-d with respective 3a or 3c, followed by oxidation and deprotection led to the isolation of the target compounds la-d in good yields.

A new system for hydrogenation of haloalkenes is reported. Cu(II)/Fe(III)-mediated selective hydrogenation of steroidal haloalkenes in the presence of hydrazine proves to be a very efficient method for the synthesis of 17β-halosteroids, potential candidates as antiestrogens or androgen receptor-mediated imaging agents. The reaction stereospecifically affords β-haloalkanes without any concomitant formation of dehalogenation products.

4-Chlorobenzoate:coenzyme A (4-CBA:CoA) ligase catalyzes 4-chlorobenzoyl-coenzyme A formation in a two-step reaction consisting of the adenylation of 4-chlorobenzoate with adenosine 5'-triphosphate followed by acyl transfer from the 4-chlorobenzoyl adenosine 5'-monophosphate diester intermediate to coenzyme A. In this study, two core motifs present in the Pseudomonas sp. strain CBS3 4-CBA:CoA ligase (motif I, 161T-S-G-T-T-G-L-P-K-G170, and motif II, 302Y-G-T-T-E306) and conserved among the sequences representing the acyl-adenylate/thioester-forming enzyme family (to which the ligase belongs) were tested for their possible role in substrate binding and/or catalysis. The site-directed mutants G163I, G166I, P168A, K169M, and E306Q were prepared and then subjected to steady-state and transient kinetic studies. The results, which indicate reduced catalysis of the adenylation of 4-chlorobenzoate in the mutant enzymes, are interpreted within the context of the three-dimensional structure of the acyl-adenylate/thioester-forming enzyme family member, firefly luciferase.

The retinoblastoma protein (Rb) plays a critical role in cell proliferation, differentiation, and development. To decipher the mechanism of Rb function at the molecular level, we have systematically characterized a number of Rb-interacting proteins, among which is the clone C5 described here, which encodes a protein of 1,978 amino acids with an estimated molecular mass of 230 kDa. The corresponding gene was assigned to chromosome 14q31, the same region where genetic alterations have been associated with several abnormalities of thyroid hormone response. The protein uses two distinct regions to bind Rb and thyroid hormone receptor (TR), respectively, and thus was named Trip230. Trip230 binds to Rb independently of thyroid hormone while it forms a complex with TR in a thyroid hormone-dependent manner. Ectopic expression of the protein Trip230 in cells, but not a mutant form that does not bind to TR, enhances specifically TR-dependent transcriptional activity. Coexpression of wild-type Rb, but not mutant Rb that fails to bind to Trip230, inhibits such activity. These results not only identify a coactivator molecule that modulates TR activity, but also uncover a role for Rb in a pathway that responds to thyroid hormone.

A gene assigned to human chromosome 1q32-41 encodes a novel protein of 3,113 amino acids containing an internal tandem repeat of 177 amino acids. The protein, which we have named "mitosin," was identified by direct binding to purified retinoblastoma protein in vitro with a region distantly related to the retinoblastoma protein-binding site of E2F-1. Mitosin is expressed throughout S, G2, and M phases of the cell cycle but is absent in G1. Its localization is dramatically reorganized from a rather homogeneous nuclear distribution in S phase to paired dots at the kinetochore/centromere region, to the spindle apparatus, and then to the midbody during M-phase progression. This spatial reorganization coincides closely with the temporal phosphorylation patterns of mitosin. Overexpression of N-terminally truncated mutants blocks cell cycle progression mainly at G2/M. These results suggest that mitosin may play an important role in mitotic-phase progression.

In eukaryotes, nuclear DNA is packaged into chromatin by binding to histones and associated factors. Covalent modifications to histone tails are associated with specific transcriptional states of the associated DNA. Acetylation of lysine side chains normally correlates with transcriptional activation, while deacetylation leads to transcriptional silencing. The regulatory roles of lysine and arginine methylation appear to be more complex. Methylation of certain lysine residues is associated with active transcription, while methylation of others is associated with silencing and heterochromatin formation. Each methylation marker is placed, removed and interpreted in a site-specific manner by histone methyltransferases, demethylases and methyl binding domains, respectively. The biological functions of the individual enzymes are largely undefined and are the focus of current investigations (for Reviews see References [1, 2]) The JmjC histone demethylases are 2-oxoglutarate (2OG)-dependent oxygenases that catalyse N-lysyl demethylation via hydroxylation of the methyl group in a 2OGand Fe-dependent manner (Scheme 1). Human 2OG oxygenases catalyse a range of reactions, including hydroxylation of amino acids, DNA, and small molecules, and demethylation of proteins and DNA. 2OG oxygenases show promise as therapeutic targets; an inhibitor of g-butyrobetaine hydroxylase (BBOX) is used for the treatment of cardiovascular disease, and inhibitors of the hypoxia inducible factor (HIF) prolyl hydroxylases are in clinical trials for the treatment of anaemia. Inhibitors of the collagen prolyl hydroxylases have also been evaluated as potential therapeutics for the treatment of liver fibrosis. 8] The discovery of the JmjC domain histone demethylases, and the suggestion that some of them are potential therapeutic targets for cancer treatment, has stimulated interest in their inhibition, but relatively few studies have been described. Reported inhibitors of the JmjC demethylases include N-oxalyl amino acids, 8-hydroxyquinolines, pyridine dicarboxylates, hydroxamic acids and catechol-type flavonoids (Figure 1). Compounds that catalyse the ejection of a structural Zn ion from the JMJD2 demethylases have also been reported (Figure 1).

Microbial enzyme systems may be used in the biodegradation of persistent environmental pollutants. The three polypeptide components of one such system, the 4-chlorobenzoate dehalogenase system, have been isolated, and the chemical steps of the 4-hydroxybenzoate-forming reaction that they catalyze have been identified. The genes contained within a 4.5-kilobase Pseudomonas sp. strain CBS3 chromosomal DNA fragment that encode dehalogenase activity were selectively expressed in transformed Escherichia coli. Oligonucleotide sequencing revealed a stretch of homology between the 57-kilodalton (kD) polypeptide and several magnesium adenosine triphosphate (MgATP)-cleaving enzymes that allowed MgATP and coenzyme A (CoA) to be identified as the dehalogenase cosubstrate and cofactor, respectively. The dehalogenase activity arises from two components, a 4-chlorobenzoate:CoA ligase-dehalogenase (an alpha beta dimer of the 57- and 30-kD polypeptides) and a thioesterase (the 16-kD polypeptide).

The three genes encoding the 4-chlorobenzene dehalogenase polypeptides were excised from a Pseudomonas sp. CBS-3 DNA fragment and separately cloned and expressed in Escherichia coli. The three enzymes were purified from the respective subclones by using an ammonium sulfate precipitation step followed by one or two column chromatographic steps. The 4-chlorobenzoate:coenzyme A ligase was found to be a homodimer (57-kDa subunit size), to require Mg2+ (Co2+ and Mn2+ are also activators) for activity, and to turn over MgATP (Km = 100 microM), coenzyme A (Km = 80 microM), and 4-chlorobenzoate (Km = 9 microM) at a rate of 30 s-1 at pH 7.5 and 25 degrees C. Benzoate, 4-bromobenzoate, 4-iodobenzoate, and 4-methylbenzoate were shown to be alternate substrates while 4-hydroxybenzoate, 4-aminobenzoate, 2-aminobenzoate, 2,3-dihydroxybenzoate, 4-coumarate, palmate, laurate, caproate, butyrate, and phenylacetate were not substrate active. The 4-chlorobenzoate-coenzyme A dehalogenase was found to be a homotetramer (30 kDa subunit size) to have a Km = 15 microM and kcat = 0.3 s-1 at pH 7.5 and 25 degrees C and to be catalytically inactive toward hydration of crotonyl-CoA, alpha-methylcrotonyl-CoA, and beta-methylcrotonyl-CoA. The 4-hydroxybenzoate-coenzyme A thioesterase was shown to be a homotetramer (16 kDa subunit size), to have a Km = 5 microM and kcat = 7 s-1 at pH 7.5 and 25 degrees C, and to also catalyze the hydrolyses of benzoyl-coenzyme A and 4-chlorobenzoate-coenzyme A. Acetyl-coenzyme A, hexanoyl-coenzyme A, and palmitoyl-coenzyme A were not hydrolyzed by the thioesterase.

Microbial enzyme systems may be used in the biodegradation of persistent environmental pollutants. The three polypeptide components of one such system, the 4-chlorobenzoate dehalogenase system, have been isolated, and the chemical steps of the 4-hydroxybenzoate-forming reaction that they catalyze have been identified. The genes contained within a 4.5-kilobase Pseudomonas sp. strain CBS3 chromosomal DNA fragment that encode dehalogenase activity were selectively expressed in transformed Escherichia coli. Oligonucleotide sequencing revealed a stretch of homology between the 57-kilodalton (kD) polypeptide and several magnesium adenosine triphosphate (MgATP)-cleaving enzymes that allowed MgATP and coenzyme A (CoA) to be identified as the dehalogenase cosubstrate and cofactor, respectively. The dehalogenase activity arises from two components, a 4-chlorobenzoate:CoA ligase-dehalogenase (an alpha beta dimer of the 57- and 30-kD polypeptides) and a thioesterase (the 16-kD polypeptide).

A new type of noncompetitive alpha-2,3-sialyltransferase inhibitor has been synthesized; we report the discovery, preparation and inhibitory activity of sixteen lithocholic acid analogues.

4-Chlorobenzoate:coenzyme A ligase (4-CBA:CoA ligase) catalyzes the first step of the 4-CBA degradation pathway of Pseudomonas sp. strain CBS3. In this reaction, 4-CBA-CoA thioester synthesis is coupled to ATP cleavage. The studies described in this paper examine the intermediacy of 4-chlorobenzoyl-adenosine 5'phosphate diester (4-CBA-AMP) in the ligase reaction. The 4-CBA-AMP adduct was isolated from the ligase reaction mixture generated from magnesium adenosine 5-triphosphate (MgATP) and 4-CBA in the absence of CoA. The structure of the 4-CBA-AMP was verified by 1H- 13C-, and 31P-nuclear magnetic resonance analysis. Single-turnover reactions carried out with 14C-labeled 4-CBA in a rapid quench apparatus demonstrated formation of the enzyme. 4-CBA-AMP.MgPPi complex from the enzyme.4-CBA.MgATP complex at a rate of 135 s-1. The rate of ligand release from the enzyme.4-CBA-AMP.MgPPi complex was measured at 0.013 s-1. Single-turnover reactions of [14C]-4-CBA, MgATP, and CoA catalyzed by the ligase revealed that the 4-CBA-AMP intermediate formed reaches a maximum level of 25% of the starting 4-CBA within 10 ms and then declines with the formation of the 4-CBA-CoA. The rates of the adenylation and thioesterification partial reactions, determined by kinetic simulation of the rate data, are nearly equal (135 and 100 s-1). Substitution of CoA with the slow substrate pantetheine did not significantly alter the rate of the adenylation step but did reduce the rate of the thioesterification step to 2 s-1. The maximum level of 4-CBA-AMP reached during the single-turnover reaction of 4-CBA, MgATP, and pantetheine corresponded to one-half of the starting 4-CBA.

We have deduced the nucleotide sequence of the genes encoding the three components of 4-chlorobenzoate (4-CBA) dehalogenase from Pseudomonas sp. CBS-3 and examined the origin of these proteins by homology analysis. Open reading frame 1 (ORF1) encodes a 30-kDa 4-CBA-coenzyme A dehalogenase related to enoyl-coenzyme A hydratases functioning in fatty acid beta-oxidation. ORF2 encodes a 57-kDa protein which activates 4-CBA by acyl adenylation/thioesterification. This 4-CBA:coenzyme A ligase shares significant sequence similarity with a large group of proteins, many of which catalyze similar chemistry in beta-oxidation pathways or in siderophore and antibiotic synthetic pathways. These proteins have in common a short stretch of sequence, (T,S)(S,G)G(T,S)(T,E)G(L,X)PK(G,-), which is particularly highly conserved and which may represent an important new class of "signature" sequence. We were unable to find any proteins homologous in sequence to the 16-kDa 4-hydroxybenzoate-coenzyme A thioesterase encoded by ORF3. Analysis of the chemistry and function of the proteins found to be structurally related to the 4-CBA:coenzyme A ligase and the 4-CBA-coenzyme A dehalogenase supports the proposal that they evolved from a beta-oxidation pathway.