Decomposition of BCNU (1,3-bis(2-chloroethyl)-1-nitrosourea) in aqueous solution

BCNU, labelled with ¹⁴C in the chloroethyl groups, decomposes in neutral aqueous solution to release half of its radioactivity as volatile products. These have been identified by a combination of gas chromatography and mass spectrometry as vinyl chloride, acetaldehyde, dichloroethane, and chloroethanol. This set of products is consistent with the existence of chloroethylcarbonium ions as reactive intermediates which would produce the previously described substituted nucleotides.     

Mechanism of action of the nitrosoureas: formation of 1,2-(diguanosin-7-yl) ethane from the reaction of BCNU (1,3-bis-[2-chloroethyl]-1-nitrosourea) with guanosine

A crosslinked dinucleoside, 1,2-(diguanosin-7-yl) ethane, has been isolated from the reaction of guanosine with the antitumor agent, BCNU. The formation of this product suggests that DNA crosslinking, which may be responsible for the cytotoxicity of BCNU, could occur through such dinucleosides.     

Structure of the single-stranded polyribonucleotide polycytidylic acid.

The Fourier transformation was obtained from the experimental diffuse X-ray scattering curve of adult human oxyhaemoglobin in an aqueous solution. The correlation function of the form of the molecule was calculated from known coordinates of horse haemoglobin atoms. The distribution function of inhomogeneities was deduced from the above Fourier transformation and the correlation function. This distribution function of inhomogeneities is described by a series of maxima providing evidence for a short and long-range order extending up to 5 nm which is close to the maximum dimension of the haemoglobin molecule.     

The synthesis ofP1-[2-acetamido-2-deoxy-3-O-(β-D-glucopyranosyluronic acid)-α-D-glucopyranosyl] P2-dolichyl diphosphate (N-acetylhyalobiosyluronic dolichyl diphosphate)

Starting from 2-acetamido-4,6-di-O-acetyl-2-deoxy-3-O-(methyl 2,3,4-tri-O-acetyl-β-D-glucopyranosyluronate)-α-D-glucopyranose (20), a crystalline intermediate prepared by a conventional sequence of reactions, the total synthesis of N-acetyl-hyalobiosyluronic dolichyl diphosphate was achieved. One of the key steps involved the transformation of the disaccharide 20 into the methyloxazoline 26, which was then converted into the stable, crystalline disaccharide phosphate derivative in ～30% yield. The methyloxazoline 26 was directly prepared from the corresponding methyl α-glycoside by acetolysis. Similarly, the allyl α-glycoside was transformed into 26.     

Chemistry of 1,1-bis(acylamido)-1-deoxyalditols. Base-catalyzed,intramolecular O,N-transacylation of per-O-acyl-1,1-bis(benzamido)-1-deoxy-d-glucitols in aprotic solvents

Penta-O-acetyl and penta-O-propanoyl derivatives of 1,1-bis(benzamido)-1-deoxy-d-glucitol are transformed into 1-acetamido-1-benzamido-1-deoxy-d-glucitol and 1-benzamido-1-deoxy-1-propanamido-d-glucitol, respectively, by heating with a suspension of potassium cyanide in acetonitrile, and subsequently O-deacylating with sodium methoxide in methanol. The reaction was also studied in the presence of a crown ether. When other nucleophiles (HO^? and CH³O^?) or other aprotic solvents (propanonitrile, benzene) were employed, the yields of transacylation products diminished noticeably; likewise, the use of sodium as the counter-ion significantly affected this reaction. These results are qualitatively discussed in terms of the solvent effects on the reactivity of the nucleophiles employed.     

Fatty acid desaturase system of yeast microsomes. Involvement of cytochrome b5-containing electron-transport chain.

The oxidative desaturation of palmitoyl CoA by microsomes from anaerobically grown Saccharomyces cerevisiae has been studied by using NADH as electron donor. The desaturation product was identified as palmitoleic acid by periodate oxidation. The desaturase activity was sensitive to relatively high concentrations of cyanide; the concentration of cyanide causing half-maximal inhibition was determined to be 7.1 mm. The rate of reoxidation of cytochrome b₅ in NADH-reduced microsomes was stimulated by the addition of palmitoyl CoA, and the amount of cytochrome b₅ reoxidized by the palmitoyl CoA added could be closely correlated to the amount of palmitoleate formed. No stimulation of the reoxidation of cytochrome b₅ was induced by palmitoyl CoA in microsomes prepared from the desaturase-repressed cells and from a desaturase-deficient mutant, strain KD-20. It is concluded that the fatty acyl CoA desaturase system of yeast microsomes involves cytochrome b₅ as an electron carrier and that the terminal desaturase is sensitive to relatively high concentrations of cyanide.     

Oligosaccharides containing fucose linked alpha(1-3) and alpha(1-4) to N-acetylglucosamine cause decompaction of mouse morulae

Two- to four-cell and eight-cell mouse embryos were incubated in various fucosylated and unfucosylated oligosaccharides, fucose binding protein, and fucosylated BSA. Compaction at the eight-cell stage was reversed by a mixture containing the oligosaccharides lacto-N-fucopentaose II (80-90%), in which fucose is linked alpha(1-4) to N-acetylglucosamine, and lacto-N-fucopentaose III (10-20%), in which fucose is linked alpha(1-3) to N-acetylglucosamine. Pure lacto-N-fucopentaose III (LNFP III) and 3-fucosyl lactose (containing fucose alpha(1-3)glucose) had a similar effect. All three molecules affected blastocyst formation. Various closely related fucosylated and unfucosylated oligosaccharides did not induce decompaction or inhibit blastocyst formation. The proportion of embryos incubated from the two- to four-cell stage in LNFP II/III which reached the eight-cell stage and formed blastocysts was reduced. Those which formed compact morulae subsequently decompacted. Precompact or early compacting eight-cell embryos incubated in LNFP II/III compacted normally but subsequently decompacted and failed to form blastocysts. Decompaction of eight-cell embryos in LNFP II/III occurred during a specific period of development (80-90 hr post-hCG) and was reversible up to 84-86 hr post-hCG, but not by 92 hr post-hCG. The period of sensitivity to LNFP II/III was associated with the decrease in the ability of calcium-free medium to cause decompaction. It appears that LNFP II/III interferes with a later calcium-independent phase of compaction and we propose that LNFP III and II inhibit an endogenous lectin-saccharide interaction between membranes involved in the stabilization of compaction.     

Free metal ion depletion by "Good's" buffers. III. N-(2-acetamido)iminodiacetic acid, 2:1 complexes with zinc(II), cobalt(II), nickel(II), and copper(II); amide deprotonation by Zn(II), Co(II), and Cu(II)

Potentiometric, visible, infrared, electron spin, and nuclear magnetic resonance studies of the complexation of N-(2-acetamido)iminodiacetic acid (H2ADA) by Ca(II), Mg(II), Mn(II), Zn(II), Co(II), Ni(II), and Cu(II) are reported. Ca(II) and Mg(II) were found not to form 2:1 ADA2- to M(II) complexes, while Mn(II), Cu(II), Ni(II), Zn(II), and Co(II) did form 2:1 metal chelates at or below physiological pH values. Co(II) and Zn(II), but not Cu(II), were found to induce stepwise deprotonation of the amide groups to form [M(H-1ADA)4-(2)]. Formation (affinity) constants for the various metal complexes are reported, and the probable structures of the various metal chelates in solution are discussed on the basis of various spectral data.     

Synthesis of p-nitrophenyl 6-O-(2-acetamido-2-deoxy-β-d-glucopyranosyl)-α-d-mannopyranoside

The reaction of p-nitrophenyl 2,3-O-isopropylidene-α-d-mannopyranoside and 2-methyl-(3,4,6-tri-O-acetyl-1,2-dideoxy-α-d-glucopyrano)-[2,1-d]-2-oxazoline gave a crystalline, 6-O-substituted disaccharide derivative which, on de-isopropylidenation followed by saponification, produced the disaccharide p-nitrophenyl 6-O-(2-acetamido-2-deoxy-β-d-glucopyranosyl)-α-d-mannopyranoside. Synthesis of methyl 6-O-(2-acetamido-2-deoxy-β-d-glucopyranosyl)-α-d-mannopyranoside was also accomplished by a similar reaction-sequence. The structures of these disaccharides have been established by ¹³C-n.m.r. spectroscopy.     

Structures of two N(1)-bound platinum(II)-6-oxopurine complexes. Comparisons with complexes derived from platinum (II) anti-tumor agents

The preparation and molecular structure of [(diethylenetriamine) (7,9-dimethylhypoxanthine) platinum(II)] (PF₆)₂·1.5H₂O and [(ethylenediamine) (7,9-dimethylhypoxanthine)₂platinum(II)] (PF₆)₂, are reported. These complexes represent the first structurally characterized N(1)-bound Pt(II) 6-oxopurine complexes. In each case, the Pt(II)N(1) bond length [2.051(6)A in the diethylenetriamine complex and 2.021(8)A in the ethylenediamine complex] indicates a strong metal-to-base binding. Both complexes contain interligand hydrogen bonds, with the ammine ligand acting as the donor and the O(6) atom of the base acting as the acceptor. These N(1)-bound complexes are compared with N(7)-bound 6-oxopurine and N(3)-bound cytosine complexes of Pt(II) anti-tumor agents.     

A new method for the detection and assay of α-1,3-glucanases

A colorimetric method that is specific for the assay of α-1,3-glucanases is presented. The enzyme substrate consists of Cibacron Blue F3GA complexed with a dextranase-treated streptococcal glucan. The method is especially convenient for tests involving large numbers of samples, and can be adapted to quantitative as well as qualitative applications. The assay is sufficiently sensitive for screening bacterial samples as potential sources of α-1,3-glucanase.     

The conversion of benzo(alpha)pyrene 4,5-oxide into 4-hydroxybenzo(alpha)pyrene in the presence of polyriboguanylic acid.

Incubation of benzo[alpha] pyrene 4,5-oxide with poly(G) in neutral aqueous ethanol resulted in the formation of covalent adducts and in the production of free 4-hydroxybenzo[alpha]pyrene. This phenol, which was identified by its UV spectral properties and by its chromatographic characteristics, was also formed but at a much slower rate when the epoxide was incubated with DNA or with GMP. Phenol formation was not detected when benzo[alpha]-pyrene 4,5-oxide was incubated for prolonged periods in the presence of poly(A), poly(C) or poly(U) or in the absence of nucleic acid. Formation of 4-hydroxybenzo[alpha] pyrene from the epoxide in the presence of poly(G) was not accompanied by detectable base modifications or by breakage of phosphodiester linkages.     

Stereoselectivity of 1-aminocyclopropanecarboxylate malonyltransferase toward stereoisomers of 1-amino-2-ethylcyclopropanecarboxylic acid

A malonyltransferase isolated from mungbean (Vigna radiata L.) hypocotyls catalyzed the malonylation of both 1-aminocyclopropane-1-carboxylic acid (ACC) and D-amino acids. The possibility that ACC was recognized by the enzyme as a D-amino acid was investigated by examining the efficiencies of the four stereoisomers of 1-amino-2-ethylcyclopropane-1-carboxylic acid (AEC) serving as substrates of malonyltransferase and as inhibitors of ACC malonyltransferase. Although all four isomers were malonylated by the enzyme and competitively inhibited the malonylation of ACC to N-malonyl-ACC, (1R,2S)-AEC and (1R,2R)-AEC, both of which have an R-configuration as a D-amino acid, had lower Km and Ki values (0.1 to 0.2 mM) than their enantiomers, (1S,2R)-AEC (Km and Ki values were about 1 mM) and (1S,2S)-AEC (Km and Ki values were higher than 10 mM), which have an S-configuration as an L-amino acid. Similarly, (R)-isovaline (2-amino-2-methylbutanoic acid), which has an R-configuration as a D-amino acid, inhibited more effectively the enzymatic conversion of ACC to malonyl-ACC than did (S)-isovaline, which has an S-configuration as an L-amino acid. In mungbean hypocotyls (1R,2S)-AEC and (1R,2R)-AEC were also more efficiently converted into malonyl conjugates and more efficiently inhibited the conversion of radioactive ACC into malonyl-ACC than their enantiomers, although the differences in efficiency among stereoisomers were smaller in hypocotyls than in enzymatic reactions. These results suggest that ACC is recognized by the enzyme as a D-amino acid.