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Biomedical subjects

S Liu

Publications and source records attributed to S Liu.

At least 271 records · Page 15Linked to original sources
PubMed2000

Trimeric ring-like structure of ArsA ATPase.

ArsA protein is the soluble subunit of the Ars anion pump in the Escherichia coli membrane which extrudes arsenite or antimonite from the cytoplasm. The molecular weight of the subunit is 63 kDa. In the cell it hydrolyzes ATP, and the energy released is used by the membrane-bound subunit ArsB to transport the substrates across the membrane. We have obtained two-dimensional crystals of ArsA in the presence of arsenite on negatively-charged lipid monolayer composed of DMPS and DOPC. These crystals have been studied using electron microscopy of negatively-stained specimens followed by image processing. The projection map obtained at 2.4 nm resolution reveals a ring-like structure with threefold symmetry. Many molecular assemblies with the same ring-shape and dimensions were also seen dispersed on electron microscopy grids, prepared directly from purified ArsA protein solution. Size-exclusion chromatography of the protein sample with arsenite present revealed that the majority of the protein particles in solution have a molecular weight of about 180 kDa. Based on these experiments, we conclude that in solution the ArsA ATPase with substrate bound is mainly in a trimeric form.

Adenosine Triphosphatases↗
PubMed2000

Carbon catabolite repression in plant pathogenic fungi: isolation and characterization of the Gibberella fujikuroi and Botrytis cinerea creA genes.

The creA genes of two plant pathogenic fungi, the gibberellin-producing rice pathogen Gibberella fujikuroi and the gray mold Botrytis cinerea, were isolated and characterized. The deduced amino acid sequences of both glucose repressors are 64% identical to each other and 59% (G. fujikuroi) and 61% (B. cinerea) identical to the CreA protein of Aspergillus nidulans. The zinc finger regions of the Gibberella and Botrytis CreA proteins shared 98% identity with the corresponding zinc finger region of the A. nidulans protein, and studies by complementation of a creA null mutant of A. nidulans showed that the proteins are functional homologues of A. nidulans CreA. Northern blot analysis revealed that creA transcript levels are independent of the carbon source in both fungi.

Amino Acid Sequence↗
PubMed2000

Class- and splice variant-specific association of CD98 with integrin beta cytoplasmic domains.

CD98 is a type II transmembrane protein involved in neutral and basic amino acid transport and in cell fusion events. CD98 was implicated in the function of integrin adhesion receptors by its capacity to reverse suppression of integrin activation by isolated integrin beta(1A) domains. Here we report that CD98 associates with integrin beta cytoplasmic domains with a unique integrin class and splice variant specificity. In particular, CD98 interacted with the ubiquitous beta(1A) but not the muscle-specific splice variant, beta(1D), or leukocyte-specific beta(7) cytoplasmic domains. The ability of CD98 to associate with integrin cytoplasmic domains correlated with its capacity to reverse suppression of integrin activation. The association of CD98 with integrin beta(1A) cytoplasmic domains may regulate the function and localization of these membrane proteins.

Amino Acid Sequence↗
PubMed2000

Homotrinuclear lanthanide(III) arrays: assembly of and conversion from mononuclear and dinuclear units.

The reactions of potentially hexadentate H2bbpen (N,N'-bis(2-hydroxybenzyl)-N,N'-bis(2-pyridylmethyl)-ethylenediamine, H2L1), H2(Cl)bbpen (N,N'-bis(5-chloro-2-hydroxybenzyl)-N,N'-bis(2-pyridylmethyl)ethylenediamine, H2L2), and H2(Br)bbpen (N,N'-bis(5-bromo-2-hydroxybenzyl)-N,N'-bis(2-pyridylmethyl)ethylenediamine, H2L3) with Ln(III) ions in the presence of a base in methanol resulted in three types of complexes: neutral mononuclear ([LnL(NO3)]), monocationic dinuclear ([Ln2L2(NO3)]+), and monocationic trinuclear ([Ln3L2(X)n(CH3OH)]+), where X = bridging (CH3COO-) and bidentate ligands (NO3-, CH3COO-, ClO4-) and n is 4. The formation of a complex depends on the base (hydroxide or acetate) and the size of the respective Ln(III) ion. All complexes were characterized by infrared spectroscopy, mass spectrometry, and elemental analyses; in some cases, X-ray diffraction studies were also performed. The structures of the neutral mononuclear [Yb(L1)(NO3)], dinuclear [Pr2(L1)2(NO3)(H2O)]NO3.CH3OH and [Gd2(L1)2(NO3)]NO3.CH3OH.3H2O, and trinuclear [Gd3(L3)2(CH3COO)4(CH3OH)]ClO4.5CH3OH and [Sm3(L1)2(CH3COO)2(NO3)2(CH3OH)]NO3.CH3OH.3.65H2O were solved by X-ray crystallography. The [LnL(NO3)] or [Ln2L2(NO3)]+ complexes could be converted to [Ln3L2(X)n(CH3OH)]+ complexes by the addition of 1 equiv of a Ln(III) salt and 2-3 equiv of sodium acetate in methanol. The trinuclear complexes were found to be the most stable of the three types, which was evident from the presence of the intact monocationic high molecular weight parent peaks ([Ln3L2(X)n]+) in the mass spectra of all the trinuclear complexes and from the ease of conversion from the mononuclear or dinuclear to the trinuclear species. The incompatibility of the ligand denticity with the coordination requirements of the Ln(III) ions was proven to be a useful tool in the construction of multinuclear Ln(III) metal ion arrays.

Contrast Media↗
PubMed2000

Influence of alcohol concentration on lipase-catalyzed enantioselective esterification of racemic naproxen in isooctane: under controlled water activity*

According to a previous study, alcohol concentration influences the enantiomeric ratio and initial rate of Candida cylindracea lipase-catalyzed enantioselective esterification of racemic 2-(6-methoxy-2-naphthyl) propionic acid in microaqueous isooctane in a similar manner. Such an influence might be attributed to the different water partitioning coefficients. In this work, we performed enzymatic resolutions in controlled water activity atmospheres, thereby separating the influence of alcohol concentrations from the effect of water partitioning. Despite this constant water activity condition, a similar dependence of enantiospecificity and initial rate on the concentration of acyl acceptor (n-butanol) was also found. This finding suggests that the alcohol concentration influences enzyme performance, but not because it strips water from the enzyme. The unexpected observations imply that an undetermined factor must be considered when the enzymatic resolution is performed in nonconventional media.

Journal Article↗
PubMed2000

Ethyl carbamate precursor citrulline formation from arginine degradation by malolactic wine lactic acid bacteria.

Major commercially available strains for induction of malolactic fermentation in wine were examined for arginine metabolism in a resting cell system at wine pH with the aim of evaluating their ability to excrete and utilize citrulline, a precursor of carcinogenic ethyl carbamate (urethane). All strains tested excreted citrulline from arginine degradation. Citrulline was stored intracellularly during growth in arginine rich medium and was released upon lysis of the cells. All strains were found to degrade citrulline as a sole amino acid and some of them were able to reutilize previously excreted citrulline.

Arginine↗
PubMed2000

Induction of apoptotic cell death and in vivo growth inhibition of human cancer cells by a saturated branched-chain fatty acid, 13-methyltetradecanoic acid.

A saturated branched-chain fatty acid, 13-methyltetradecanoic acid (13-MTD), was purified from a soy fermentation product, which was used by many cancer patients as a treatment supplement. Our preliminary study indicated that 13-MTD could induce cell death in human cancer cell lines K-562, MCF7, DU 145, NCI-SNU-1, SNU-423, NCI-H1688, BxPC3, and HCT 116. The ID50 dosage of 13-MTD for these tumor cells ranged from 10 to 25 microg/ml. Further investigation revealed that 13-MTD caused tumor cell death through rapid induction of apoptosis, which could be detected 2 h after the treatment of tumor cells with 13-MTD. Xenograft tumors of prostate carcinoma cell line DU 145 and hepatocarcinoma LCI-D35 were orthotopically implanted into nude mouse prostate and liver, respectively. 13-MTD was administered p.o. once daily to the implanted mice for approximately 40 days. Our results showed that 13-MTD could effectively inhibit the growth of orthotopic tumor implants of both cell lines compared with control groups. The average inhibition rate was 84.6% for DU 145 and 65.2% for LCI-D35 (P < 0.01). LD50 test results showed that mice could well sustain the oral feeding of 5 g/kg/day without observable anomaly. Our preliminary data demonstrated that 13-MTD could effectively inhibit in vitro and in vivo growth of various cancer cell lines by inducing apoptosis without significant toxic side effects, suggesting 13-MTD as a potential candidate for chemotherapy of human cancers.

Animals↗
PubMed2000

Structures of human dihydroorotate dehydrogenase in complex with antiproliferative agents.

BACKGROUND: Dihydroorotate dehydrogenase (DHODH) catalyzes the fourth committed step in the de novo biosynthesis of pyrimidines. As rapidly proliferating human T cells have an exceptional requirement for de novo pyrimidine biosynthesis, small molecule DHODH inhibitors constitute an attractive therapeutic approach to autoimmune diseases, immunosuppression, and cancer. Neither the structure of human DHODH nor any member of its family was known. RESULTS: The high-resolution crystal structures of human DHODH in complex with two different inhibitors have been solved. The initial set of phases was obtained using multiwavelength anomalous diffraction phasing with selenomethionine-containing DHODH. The structures have been refined to crystallographic R factors of 16.8% and 16.2% at resolutions of 1. 6 A and 1.8 A for inhibitors related to brequinar and leflunomide, respectively. CONCLUSIONS: Human DHODH has two domains: an alpha/beta-barrel domain containing the active site and an alpha-helical domain that forms the opening of a tunnel leading to the active site. Both inhibitors share a common binding site in this tunnel, and differences in the binding region govern drug sensitivity or resistance. The active site of human DHODH is generally similar to that of the previously reported bacterial active site. The greatest differences are that the catalytic base removing the proton from dihydroorotate is a serine rather than a cysteine, and that packing of the flavin mononucleotide in its binding site is tighter.

Amino Acid Sequence↗
PubMed2000

Patterns of infant mortality caused by major congenital anomalies.

BACKGROUND: We assessed the impact of recent advances in perinatal care on infant mortality due to congenital anomaly. METHODS: Analysis of trends in congenital anomaly-attributed infant mortality, using the 1981-1995 Statistics Canada's birth and death records, with a total of 2,878,826 live births, 21,883 infant deaths, and 6, 908 infant deaths due to congenital anomalies. RESULTS: Infant mortality due to major congenital anomaly decreased from 3.11 per 1, 000 live births in 1981 to 1.89 per 1,000 live births in 1995. Cause-specific infant mortality rates for anencephaly, spina bifida, other central nervous system anomalies, cardiovascular system anomalies, respiratory system anomalies, digestive system anomalies, certain musculoskeleton anomalies, urinary system anomalies, chromosomal anomalies, and multiple congenital anomalies were 0.20, 0.23, 0.27, 1.04, 0.24, 0.08, 0.22, 0.16, 0.22, and 0.13 per 1,000 live births, respectively, in 1981-1983, whereas corresponding rates were 0.07, 0.07, 0.18, 0.73, 0.25, 0.03, 0.12, 0.12, 0.26, and 0.06 per 1,000 live births, respectively, in 1993-1995. CONCLUSIONS: Recent Canadian data show that infant deaths caused by major congenital anomalies have decreased significantly, but reductions varied substantially according to specific forms of anomalies.

Canada↗
PubMed2000

Hydrogen-deuterium exchange reactions of cis- and trans-cyclopropane derivatives with D(2)O and CD(3)OD in the gas phase

Gas-phase hydrogen-deuterium (H/D) exchange reactions involving four isomeric cyclopropane derivatives were investigated under chemical ionization (CI) conditions, using D(2)O and CD(3)OD as reagent gases. There are abundant ions at [M + 1](+), [M + 2](+) and [M + 3](+) in the D(2)O and CD(3)OD positive-ion CI mass spectra of the two isomer pairs 1, 2 and 3, 4. Their CI mass spectra are identical with each pair, and so are the collision-induced dissociation (CID) spectra of ions [M + 1](+), [M + 2](+) and [M + 3](+) of each of the two isomer pairs. The CID spectra of [M + 1](+) ions indicate that they have common D/H exchange reactions within each pair, which take place between molecular ions and deuterium-labeling reagents to form the [M - H + D](+) ions. Those of their [M + 2](+) ions show that they have common D/H exchange reactions within each pair, which form the [M(d1) + H](+) ions. Those of their [M + 3](+) ions show that they have common D/H exchange reactions within each pair, which take place between the [M(d1)] and deuterium-labeling reagents to produce [M(d2) + H](+) for the isomer pair 1, 2 and [M(d1) + D](+) for the isomer pair 3, 4. The number and position, and active order of the active hydrogen atoms of the isomer pairs 1, 2 and 3, 4 were determined. Copyright 2000 John Wiley & Sons, Ltd.

Journal Article↗
PubMed2000

Analysis of oxosteroids by nano-electrospray mass spectrometry of their oximes.

A method for the analysis of neutral oxosteroids by electrospray mass spectrometry is described. The oxosteroids are converted into their oximes by treatment with hydroxyammonium chloride in aqueous methanol. Intense peaks corresponding to protonated oxime molecules are observed in nano-electrospray mass spectra. The detection limits for the oximes of progesterone, pregnenolone and dehydroepiandrosterone were 2.5, 5 and 25 pg/microL, respectively, approximately 20 times lower than for the underivatised steroids. The signal intensities were proportional to the concentration of the steroids in the range of 500 to 2.5 pg/microL. Fragmentation by collision-induced dissociation (CID) was studied using oximes of 28 model steroids carrying an oxo group at C-3, C-17 or C-20. Some of the steroid oximes were labelled with deuterium or (15)N. Fragment ions were observed which yielded useful structural information. Upon CID, protonated oximes of 3-oxo-Delta(4)-steroids produced abundant ions by cleavage through the B-ring and by loss of the side chain, while protonated oximes of saturated 3-oxosteroids did not give abundant ions by cleavage through the B-ring. Protonated oximes of 20-oxosteroids unsubstituted at C-21, C-17 or C-16 produced a characteristic ion at m/z 86 containing the side chain, C-16 and C-17. Protonated oximes of steroids containing only a 17-oxo group gave fewer ions of diagnostic value. Coupled with the selective isolation of steroid oximes from a biological matrix this method of derivatisation and CID may be used for the analysis of neutral oxosteroids in biological samples.

Dehydroepiandrosterone↗
PubMed2000

Capillary electrophoresis on microchip.

Capillary electrophoresis and related techniques on microchips have made great strides in recent years. This review concentrates on progress in capillary zone electrophoresis, but also covers other capillary techniques such as isoelectric focusing, isotachophoresis, free flow electrophoresis, and micellar electrokinetic chromatography. The material and technologies used to prepare microchips, microchip designs, channel geometries, sample manipulation and derivatization, detection, and applications of capillary electrophoresis to microchips are discussed. The progress in separation of nucleic acids and proteins is particularly emphasized.

Animals↗
PubMed2000

Rapid identification of saponins in plant extracts by electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/tandem mass spectrometry.

Electrospray ionization multi-stage tandem mass spectrometry (ESI-MS(n)) and liquid chromatography coupled with on-line mass spectrometry (LC/MS/MS) were applied to characterize saponins in crude extracts from Panax ginseng. The MS(n) data of the [M - H](-) ions of saponins can provide structural information on the sugar sequences of the saccharide chains and on the sapogins of saponins. By ESI-MS(n), non-isomeric saponins and isomeric saponins with different aglycones can be determined rapidly in plant extracts. LC/MS/MS is a good complementary analytical tool for determination of isomeric saponins. These approaches constitute powerful analytical tools for rapid screening and structural assignment of saponins in plant extracts.

Chromatography, Liquid↗
PubMed2000

A study of resonance electron capture ionization on a quadrupole tandem mass spectrometer.

Procedures that allow the realization of resonance electron capture (REC) mode on a commercial triple-quadrupole mass spectrometer, after some simple modifications, are described. REC mass spectrometry (MS) and tandem mass spectrometry (MS/MS) experiments were performed and spectra for some compounds were recorded. In particular, the charge-remote fragmentation (CRF) spectra of [M - H](-) ions of docosanoic and docosenoic acids under low-energy collisionally activated dissociation (CAD) conditions were obtained, and showed that there were no significant differences for [M - H](-) ions produced at different resonances (i.e. for [M - H](-) ions with different structures). This observation was explained on the basis of results obtained from deuterium-labeled fatty acids, which showed that different CRF ions (but with the same m/z value in the absence of labels) could be produced by different mechanisms, and all of them were obviously realized under CAD conditions that made spectra practically indistinguishable. The other example, which compared the REC-MS/MS spectrum of [M - H](-) ions and EI-MS/MS spectrum of M(+.) ions of daidzein, demonstrated the potential of the REC-MS/MS technique for more complex structure elucidation.

Estrogens, Non-Steroidal↗
PubMed2000

Improvement in the yield of lipophilized lysozyme by the combination with Maillard-type glycosylation.

Hen egg white lysozyme was modified using the Maillard-type glycosylation method prior to the lipophilization with palmitic acid. The yield of lipophilized lysozyme significantly increased by the pre-glycosylation of the protein. The lipophilized lysozyme derivative was separated into two main fractions with different level of glycosylation. All fractions showed a strong antimicrobial activity against Gram-negative bacteria, Escherichia coli. The lipophilization of the lysozyme combined with glycosylation is a promising method for potential industrial applications of the lysozyme due to the enhanced antimicrobial activity and the improved yield.

Anti-Bacterial Agents↗
PubMed2000

cAMP induces CD14 expression in murine macrophages via increased transcription.

CD14, a glycoprotein that binds bacterial lipopolysaccharide, plays a critical role in the inflammatory response to infection by gram-negative bacteria. Studies were undertaken to determine whether cyclic adenosine monophosphate (cAMP) regulates CD14 expression in macrophages. Incubation of RAW 264.7 cells with 8-Br-cAMP resulted in a significant increase in steady-state CD14 mRNA levels. The increase in mRNA levels was also associated with both cell-associated and soluble CD14 protein. H89 completely blocked the 8-Br-cAMP-induced CD14 mRNA up-regulation. There was no change in CD 14 mRNA half-life in the presence of 8-Br-cAMP. The CD14 gene transcription rate was increased about twofold after exposure to 8-Br-cAMP. cAMP-dependent increases in CD14 mRNA were also observed in rat peritoneal macrophages, demonstrating that this is an authentic response of mature macrophages. This study provides evidence that cAMP and protein kinase A are important regulators of CD14 expression in macrophages.

1-Methyl-3-isobutylxanthine↗
PubMed2000

Life and death decisions in B1 lymphoma cells.

Crosslinking of surface immunoglobulin (Ig) receptors with anti-IgM (anti-mu) but not anti-IgD (anti-delta) antibodies causes growth arrest and apoptosis in several extensively characterized B1-like lymphoma cell lines. While anti-mu stimulates a transient increase in c-myc mRNA and protein expression, followed by a rapid decline below the baseline level, anti-delta only causes a moderate increase in the expression of this oncogene, which returns to baseline levels within 24-48 hours. However, signals downstream from anti-delta can be converted into an apoptotic pathway by modulating PI3K activity, suggesting that PI3K is a critical rheostat controlling survival signals in B1 cell lines. Anti-mu-induced down-regulation of c-Myc is followed in time with an increase in the cyclin dependent kinase inhibitor, p27Kip1, in all anti-mu sensitive lymphoma lines. This increase correlates with growth arrest and apoptosis. The anti-mu-mediated decrease in c-Myc, increase in p27Kip1, growth arrest and apoptosis, can all be prevented via CD40/CD40L signaling. Inhibition of caspase activation, on the other hand, prevents anti-mu-induced apoptosis, but has no effect on c-Myc, p27Kip1, and G1 arrest. Interestingly, we also found that steroids and retinoids can mimic anti-mu-mediated signaling and lead to a loss of c-Myc, an increase in p27Kip1, G1 arrest, and apoptosis. Together, these data suggest that modulation of c-Myc and p27Kip1 protein levels is crucial for the life versus death decisions in murine immature B1-like lymphoma cells lines.

Antibodies, Anti-Idiotypic↗