Sugar-Induced Chiral Orientation of Boronic-Acid-Appended Amphiphile Clusters Formed in the Dipalmitoylphosphatidylcholine (DPPC) Membrane

A boronic-acid-appended amphiphile bearing an azobenzene chromophore in the chain center ( 3 ) was synthesized. Although 3 could not form the membrane-like, ordered aggregate by itself, it formed a phase-separated aggregate in a dipalmitoylphosphatidylcholine (DPPC) matrix membrane. When saccharides were added, the boronic acid group reversibly formed the saccharide complexes, and 3 in the DPPC matrix membrane became CD-active with the appearance of exciton-coupling bands. Comparison of the saccharide absolute configuration with the CD intensity established that the saccharide possessing the OH group (as 3-OH, 4-OH, and 5-CH₂OH) in the same side as the cis-1,2-diol gives the strong CD band. Judging from the structure of 3 –saccharide complexes, these “same-side” OH groups can enjoy intermolecular hydrogen-bonding interactions, which eventually induce the chiral orientation of azobenzene chromophores. This is a novel membrane system useful to read out the information stored in the saccharide structure and to create novel membrane structures by added saccharides.     

Substitutions of alanine for cysteine at a reactive thiol site and for lysine at a pyridoxal phosphate binding site of 1-aminocyclopropane-1-carboxylate deaminase

1-Aminocyclopropane-1-carboxylate (ACC) deaminase catalyzes the cyclopropane ring fragmentation and deamination of ACC. Replacement of cysteine with alanine at a reactive thiol site, Cys-162, of ACC deaminase did not affect the enzyme activity, in spite of the previous result that modification of Cys-162 caused complete loss of the enzyme activity. Substitution of glycine or valine for the cysteine residue gave a higher Km for ACC without a significant change of the K0, indicating that changes of the amino acid side chain had structural effects on substrate binding. Replacement of lysine with alanine at the pyridoxal phosphate (PLP) binding site of the ACC deaminase caused a lower content of PLP and loss of detectable activity of ACC deamination. This mutant enzyme, K51A, showed absorption peaks at 330 nm and 405 nm. The peak at 405 nm was shifted to about 425 nm by the addition of ACC, D-, L-alanine, and D-, L-serine. The formation of aldimine complexes indicated by the spectral shift was reversible. It is suggested that lysine 51 affects the formation of holoenzyme and is important in catalysis.     

Illegitimate recombination leading to allelic loss and unbalanced translocation in p53-mutated human lymphoblastoid cells

Allelic loss and translocation are critical mutational events in human tumorigenesis. Allelic loss, which is usually identified as loss of heterozygosity (LOH), is frequently observed at tumor suppressor loci in various kinds of human tumors. It is generally thought to result from deletion or mitotic recombination between homologous chromosomes. In this report, we demonstrate that illegitimate (nonhomologous) recombination strongly contributes to the generation of allelic loss in p53-mutated cells. Spontaneous and X-ray-induced LOH mutations at the heterozygous thymidine kinase (tk) gene, which is located on the long arm of chromosome 17, from normal (TK6) and p53-mutated (WTK-1) human lymphoblastoid cells were cytogenetically analyzed by chromosome 17 painting. We observed unbalanced translocations in 53% of LOH mutants spontaneously arising from WTK-1 cells but none spontaneously arising from TK6 cells. We postulate that illegitimate recombination was occurring between nonhomologous chromosomes after DNA replication, leading to allelic loss and unbalanced translocations in p53-mutated WTK-1 cells. X-ray irradiation, which induces DNA double-strand breaks (DSBs), enhanced the generation of unbalanced translocation more efficiently in WTK-1 than in TK6 cells. This observation implicates the wild-type p53 protein in the regulation of homologous recombination and recombinational DNA repair of DSBs and suggests a possible mechanism by which loss of p53 function may cause genomic instability.     

Determinant analysis of IgE and IgG4 antibodies and T cells specific for bovine alpha(s)1-casein from the same patients allergic to cow's milk: existence of alpha(s)1-casein-specific B cells and T cells characteristic in cow's-milk allergy

In an effort to clarify the etiology of milk allergy from the standpoint of allergen-specific immune reactions, we investigated the determinants of IgE, IgG4, and T cells specific for bovine alpha(s)1-casein from the same individual patients by using its synthetic peptides and cyanogen bromide-digested fragments. Alpha(s)1-casein is a major allergen in cow's milk, and its unique conformation enabled us to investigate the determinants of antibodies without consideration about missing the reactivities because of conformational changes. Nine patients were selected as subjects from among 129 milk-sensitive infants screened by ELISA to assess the anti-alpha(s)1-casein IgE levels in their sera. By using ELISA for epitope mapping, a C-terminal region of alpha(s)1-casein was identified as a common binding site for IgE from all of these patients, whereas those for anti-alpha(s)1-casein IgG4 were located in multiple regions of alpha(s)1-casein. We determined the specificities of seven alpha(s)1-casein-specific T-cell lines established from peripheral blood mononuclear cells of two of the patients. These T cells have been shown to secrete IL-4. All of the T-cell lines had different specificities to alpha(s)1-casein. However, a common amino acid residue use was found among the determinants of various T-cell lines from each patient. The results suggest that patients allergic to cow's milk have characteristic B cells recognizing a limited region of alpha(s)1-casein and secreting alpha(s)1-casein-specific IgE. These B cells may interact particularly with T cells recognizing determinants with a common structure.     

Biosensing Properties of TitanateNanotube Films: Selective Detection of Dopamine in the Presence of Ascorbate and Uric Acid

A novel strategy based on titanate nanotubes (TNTs) for developing an electrochemical biosensor is proposed. Stable TNT films are fabricated on glassy carbon (GC) electrodes by a casting technique. Cyclic voltammetry, electrochemical impedance spectrometry, and linear‐sweep voltammetry are used to characterize the TNT membrane‐covered GC electrodes (TNT/GCs). The TNT film is shown to demonstrate selectivity by charge exclusion. The TNT film is also shown to be capable of improving the mass transport to the electrode surface and electron transfer between dopamine (DA) and the electrode. Therefore, DA exhibits a quasireversible electrochemical reaction at the TNT/GC electrode. The voltammetric signal of DA is well resolved from those of ascorbate (AA) and uric acid (UA) at the TNT/GC electrode; therefore, DA can be selectively detected in the presence of a large excess of AA and UA at physiological pH. The linear calibration curve for DA is obtained over the concentration range 0.1–30 μM in a physiological solution that contains 0.1 mM AA and 0.3 mM UA.     

Calculation of the interfacial properties of liquid steel – slag systems

The rates of many high temperature metallurgical reactions are strongly influenced by interfacial phenomena and to determine reaction rates it is necessary to have an accurate model of interfacial properties. In this article, the factors governing interfacial tension in slag-metal systems will be briefly discussed and, a model based on the combination of the Gibbs and Langmuir adsorption isotherms with Girifalco and Good's expression for the interfacial tension between immiscible liquids, will be shown to be consistent with current literature values and allow the calculation of both the interfacial tension and the contact angle between liquid slags and liquid iron in the CaO–CaF₂–SiO₂–Al₂O₃ system. The application of this approach to systems containing FeO in the slag and carbon in the metal are then discussed.     

19-Allylaminoherbimycin A, an analog of herbimycin A that is stable against treatment with thiol compounds or granulocyte-macrophage colony-stimulating factor in human leukemia cells

OBJECTIVE: To investigate the effects on lipid and lipoprotein metabolism of two doses (5- or 10 micrograms/24 h) of levonorgestrel released from an intrauterine device (IUD) in combination with orally administered estradiol (2 mg estradiol valerate) in perimenopausal women. DESIGN: A 1-year prospective randomized single blind clinical trial. SETTING: Department of Obstetrics and Gynaecology, Ostra Hospital, G?teborg, Sweden. SUBJECTS: Fifty-one perimenopausal women with climacteric symptoms. OUTCOME MEASURES: Cholesterol in serum and in lipoprotein fractions; high-density lipoprotein (HDL), low-density lipoprotein (LDL). Triglycerides in serum and in very low-density lipoprotein. RESULTS: In both treatment groups significant elevations in HDL-cholesterol of similar magnitude were observed after 1 month and these changes were maintained during the 12 month observation period. In both treatment groups an initial significant decrease of LDL-cholesterol was observed and the decrement was maintained after 12 months. Serum levels of cholesterol decreased significantly in both groups after 1 month and were maintained after 12 months in the levonorgestrel-IUD (LNG-IUD) 5 micrograms group. However, the initial reduction of serum cholesterol in the LNG-IUD 10 micrograms group did not differ from baseline after 12 months. Serum triglyceride levels fluctuated during the observation period. No significant changes occurred. CONCLUSION: Continuous combined HRT with intrauterine administration of levonorgestrel, 5- or 10 micrograms/24 h, in perimenopausal women was observed to increase HDL-cholesterol and to decrease LDL-cholesterol compared with pretreatment values. The low doses of levonorgestrel did not reverse the beneficial effects on lipid metabolism usually seen after estradiol administration.