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K1
Kinds of K1 Selected AbstractsGenetic toxicity of methamphetamine in vitro and in human abusersENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 4 2003Jih-Heng Li Abstract Methamphetamine (METH) is a widely abused psychomotor stimulant. Although numerous studies have examined METH-induced neurotoxicity, its ability to produce genotoxic effects has not been evaluated. In this article, we report on the genotoxicity of METH in vitro and in human METH abusers. METH induced his+ revertants in Salmonella typhimurium strains TA98 and TA100, and increased the frequency of hprt mutants, micronuclei, and sister chromatid exchange (SCE) in cultured Chinese hamster ovary K1 (CHO-K1) cells. These METH-induced genotoxic effects were eliminated if METH exposure was conducted in the presence of rat liver S9, indicating that the genotoxicity was caused by METH, and not by metabolites of METH. In addition, reactive oxygen species (ROS) scavengers inhibited the METH-induced micronuclei in CHO-K1 cells. Further investigation with 76 human long-term METH abusers and 98 unexposed controls demonstrated that total METH exposure correlated with micronucleus and SCE frequencies in cultured lymphocytes. The results of this study indicate that METH is a genotoxic agent and that ROS may play a role in METH-induced genotoxicity. Environ. Mol. Mutagen. 42:233,242, 2003. © 2003 Wiley-Liss, Inc. [source] Characteristics of okadaic acid,induced cytotoxic effects in CHO K1 cellsENVIRONMENTAL TOXICOLOGY, Issue 6 2003C. Huynh-Delerme Abstract This article reports the results of investigations into the process of cell death induced in the Chinese hamster ovary cell K1 subclone (CHO K1) by okadaic acid (OA), a hydrophobic polyether produced by marine dinoflagellates. The IC50 was about 13 nM OA after 24 h of treatment, as determined using neutral red. With the MTT assay, the IC50 was 25 nM, although in this case 25% of the initial staining was still observed at 100 nM. Hoechst staining showed that mitotic figures accumulated at 12 nM OA after a 24- or 48-h treatment. In experiments limited to a 3-day treatment without changing the medium, CHO K1 cells were engaged in the death process at 50 nM OA after about 20 h and at 10 nM OA after 48 h. In many cells nuclear fragmentation that resulted in the apparent appearance of vesicles correlated with increasing cellular volume. But additional cell fragmentation was not observed with any treatment, and the chromatin material seemed to progressively disappear inside the cells. DNA fragmentation was analyzed by electrophoresis and with the TUNEL technique. With both techniques, the DNA was fragmented by 48 h in both 25 and 50 nM OA. Electrophoresis showed that both adherent and nonadherent cells were affected. Annexin-positive/ propidium iodide (PI),negative cells were rarely observed after OA treatment. Some were seen under the scanning cytometer after 20 h at 50 nM OA or after 48 h at 10 nM OA, but they were never detected by flow cytometry. Most of the time scanning cytometry showed either unstained cells or PI-positive (annexin-positive or -negative) cells (48 h, 50 nM, or 72 h, 10 nM). Flow cytometry cytograms showed two cell subpopulations: one composed of a majority of smaller cells, the other of larger cells. The larger cells markedly decreased with time and OA treatment (50 and 100 nM). Stained-cell counting showed that all cells that stained were both annexin- and PI positive and that most PI-positive cells were smaller. Ki67 antigen labeling showed the proliferative activity of CHO K1 cultures but also demonstrated the loss of this activity in smaller cells treated with 50 nM OA for 48 h. We concluded that in our culture conditions the main OA target within CHO K1 cultures was dividing cells. Our results suggest that cells with disturbed metaphase,anaphase enter apoptosis, leading to necrotic daughter cells. © 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 383,394, 2003 [source] Structural, Photophysical and Chiro-Optical Properties of Lanthanide Complexes with a Bis(benzimidazole)pyridine-Based Chiral LigandEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 22 2003Gilles Muller Abstract The neutral LnIII 1:1 nitrato complexes with the chiral ligand 2,6-bis(1- S -neopentylbenzimidazol-2-yl)pyridine (L11) have been synthesised and their stability constants measured in acetonitrile (log K1 = 4.0,6.4). The crystal and molecular structure of [Eu(NO3)3(L11)(MeCN)] shows the typical meridional planar coordination of L11 to the metal ion and low symmetry of the coordination polyhedron. The influence of the steric hindrance generated by the substituent at R2 on the crystal packing and bond lengths is discussed. Photophysical measurements show that ligand L11 induces a 3,,*-to-Ln energy-transfer process in the EuIII complex, while the TbIII compound is ten times less luminescent. Addition of a second molecule of L11 to give [Ln(ClO4)2(L11)2]+ leads to a large quenching of the EuIII luminescence (140-fold) due to several factors: a less efficient 1,,*,3,,* transfer (ca. fourfold), a smaller intrinsic quantum yield QEu (ca. threefold), and a substantially less efficient ligand-to-metal transfer (ca. 12-fold). In the case of the TbIII complex, the decrease in the energy of the triplet state reduces further the TbIII emission through increased back transfer. The specific rotary dispersion of the 1:1 and 1:2 complexes points to the chirality of the complexes arising mainly from the ligand, while the circularly polarized luminescence of these complexes with EuIII and TbIII displays a weak effect, pointing to a small diastereomeric excess in solution. Altogether, this study demonstrates that electronic, thermodynamic and photophysical properties of lanthanide complexes with aromatic terdentate ligands can be tuned by modifying the number and the arrangement of the ligands, as well as their substituents, particularly those in the R2 and R3 positions. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003) [source] Identification of a novel mutation in keratin 1 in a family with epidermolytic hyperkeratosisEXPERIMENTAL DERMATOLOGY, Issue 1 2000M. J. Arin Abstract: Epidermolytic hyperkeratosis (EHK) is a hereditary skin disorder typified by blistering due to cytolysis. One in 100,000 individuals is affected by this autosomal-dominant disease. The onset of the disease phenotype is typically at birth. Histological and ultrastructural examination of the epidermis shows a thickened stratum corneum and tonofilament clumping around the nucleus of suprabasal keratinocytes. Linkage studies localized the disease genes on chromosomes 12q and 17q which contain the type II and type I keratin gene clusters. Recently, several point mutations in the genes encoding the suprabasal keratins, K1 and K10, have been reported in EHK patients. We have investigated a large kindred affected by EHK and identified a new point mutation in the 2B region of keratin 1 (I107T), resulting from a T to C transition in codon 478. [source] Kinetic study of sn -glycerol-1-phosphate dehydrogenase from the aerobic hyperthermophilic archaeon, Aeropyrum pernix K1FEBS JOURNAL, Issue 3 2002Jin-Suk Han A gene having high sequence homology (45,49%) with the glycerol-1-phosphate dehydrogenase gene from Methanobacterium thermoautotrophicum was cloned from the aerobic hyperthermophilic archaeon Aeropyrum pernix K1 (JCM 9820). This gene expressed in Escherichia coli with the pET vector system consists of 1113 nucleotides with an ATG initiation codon and a TAG termination codon. The molecular mass of the purified enzyme was estimated to be 38 kDa by SDS/PAGE and 72.4 kDa by gel column chromatography, indicating presence as a dimer. The optimum reaction temperature of this enzyme was observed to be 94,96 °C at near neutral pH. This enzyme was subjected to two-substrate kinetic analysis. The enzyme showed substrate specificity for NAD(P)H- dependent dihydroxyacetone phosphate reduction and NAD+ -dependent,glycerol-1-phosphate (Gro1P) oxidation. NADP+ -dependent Gro1P oxidation was not observed with this enzyme. For the production of Gro1P in A. pernix cells, NADPH is the preferred coenzyme rather than NADH. Gro1P acted as a noncompetitive inhibitor against dihydroxyacetone phosphate and NAD(P)H. However, NAD(P)+ acted as a competitive inhibitor against NAD(P)H and as a noncompetitive inhibitor against dihydroxyacetone phosphate. This kinetic data indicates that the catalytic reaction by glycerol- 1-phosphate dehydrogenase from A. pernix follows a ordered bi,bi mechanism. [source] The Hek outer membrane protein of Escherichia coli is an auto-aggregating adhesin and invasinFEMS MICROBIOLOGY LETTERS, Issue 2 2007Robert P. Fagan Abstract Escherichia coli is the principal gram-negative causative agent of sepsis and meningitis in neonates. The pathogenesis of meningitis due to E. coli K1 involves mucosal colonization, transcytosis of epithelial cells, survival in the blood stream and eventually invasion of the meninges. The latter two aspects have been well characterized at a molecular level in the last decade. Less is known about the early stages of pathogenesis, i.e. adhesion to and invasion of gastrointestinal cells. Here, the characterization of the Hek protein is reported, which is expressed by neonatal meningitic E. coli (NMEC) and is localized to the outer membrane. It is demonstrated that this protein can cause agglutination of red blood cells and can mediate autoaggregation. Escherichia coli expressing this protein can adhere to and invade epithelial cells. So far, this is the first outer membrane protein in NMEC to be directly implicated in epithelial cell invasion. [source] Kinetics of the simultaneous oxidation of nickel(II) and sulfur(IV) by oxygen in alkaline medium in Ni(II),sulfur(IV),O2 systemINTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 8 2010Anil Kumar Sharma In the Ni(II),S(IV),O2 system in the region of pH > 8.4, both Ni(II) and S(IV) are simultaneously autoxidized, and when sulfur is consumed fully NiOOH precipitates. At pH > 8.4, ethanol has no effect on the rate, whereas ammonia strongly inhibits the reaction when pH > 7.0. The kinetics of the reaction, in both the presence and the absence of ethanol, is defined by the rate law where k is the rate constant, KO is the equilibrium constant for the adsorption of O2 on Ni(OH)2 particle surface. In ammonia buffer, the factor F is defined by where K, KOH, K1, K2, K3, and K4 are the stability constants of NiSO3, NiOH+, Ni(NH3)2+, Ni(NH3), Ni(NH3), and Ni(NH3), respectively. In unbuffered medium, the factor F reduces to The values of k and Ksp were found to be (1.3 ± 0.08) × 10,1 s,1 and (4.2 ± 3.5) × 10,16, respectively, at 30°C. A nonradical mechanism that assumes the adsorption of both SO32, and O2 on the Ni(OH)2 particle surface has been proposed. At pH , 8.2, Ni(II) displays no catalytic activity for sulfur(IV)-autoxidation and it is also not oxidized to NiOOH. © 2010 Wiley Periodicals, Inc. Int J Chem Kinet 42: 464,478, 2010 [source] Kinetics and mechanism of oxidation of the drug mephenesin by bis(hydrogenperiodato)argentate(III) complex anionINTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 8 2007Shigang Shen Mephenesin is being used as a central-acting skeletal muscle relaxant. Oxidation of mephenesin by bis(hydrogenperiodato)argentate(III) complex anion, [Ag(HIO6)2]5,, has been studied in aqueous alkaline medium. The major oxidation product of mephenesin has been identified as 3-(2-methylphenoxy)-2-ketone-1-propanol by mass spectrometry. An overall second-order kinetics has been observed with first order in [Ag(III)] and [mephenesin]. The effects of [OH,] and periodate concentration on the observed second-order rate constants k, have been analyzed, and accordingly an empirical expression has been deduced: k, = (ka + kb[OH,])K1/{f([OH,])[IO,4]tot + K1}, where [IO,4]tot denotes the total concentration of periodate, ka = (1.35 ± 0.14) × 10,2M,1s,1 and kb = 1.06 ± 0.01 M,2s,1 at 25.0°C, and ionic strength 0.30 M. Activation parameters associated with ka and kb have been calculated. A mechanism has been proposed to involve two pre-equilibria, leading to formation of a periodato-Ag(III)-mephenesin complex. In the subsequent rate-determining steps, this complex undergoes inner-sphere electron transfer from the coordinated drug to the metal center by two paths: one path is independent of OH, whereas the other is facilitated by a hydroxide ion. In the appendix, detailed discussion on the structure of the Ag(III) complex, reactive species, as well as pre-equilibrium regarding the oxidant is provided. © 2007 Wiley Periodicals, Inc. Int J Chem Kinet 39: 440,446, 2007 [source] Kinetics and mechanism of oxidation of a ternary complex involving dipicolinatochromium(III) and DL -aspartic acid by N -bromosuccinimideINTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 7 2004Hassan A. Ewais The kinetics of oxidation of [CrIII(Dpc)(Asp)(H2O)2] (Dpc = dipicolinic acid and Asp = DL -aspartic acid) by N -bromosuccinimide (NBS) in aqueous solution have been found to obey the equation: where k2 is the rate constant for the electron transfer process, K1 is the equilibrium constant for deprotonation of [CrIII(Dpc)(Asp)(H2O)2], K2 and K3 are the pre-equilibrium formation constants of precursor complexes [CrIII(Dpc)(Asp)(H2O)(NBS)] and [CrIII(Dpc)(Asp)(H2O)(OH)(NBS)],. Values of k2 = 4.85 × 10,2 s,1, K1 = 1.85 × 10,7 mol dm,3, and K2 = 78.2 mol,1 dm3 have been obtained at 30°C and I = 0.1 mol dm,3. The experimental rate law is consistent with a mechanism in which the deprotonated [CrIII(Dpc)(Asp)(H2O)(OH)], is considered to be the most reactive species compared to its conjugate acid. It is assumed that electron transfer takes place via an inner-sphere mechanism. © 2004 Wiley Periodicals, Inc. Int J Chem Kinet 36: 394,400, 2004 [source] Rate constants for the gas-phase reaction of CF3CF2CF2CF2CF2CHF2 with OH radicals at 250,430 KINTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 1 2004L. Chen The rate constants k1 for the reaction of CF3CF2CF2CF2CF2CHF2 with OH radicals were determined by using both absolute and relative rate methods. The absolute rate constants were measured at 250,430 K using the flash photolysis,laser-induced fluorescence (FP-LIF) technique and the laser photolysis,laser-induced fluorescence (LP-LIF) technique to monitor the OH radical concentration. The relative rate constants were measured at 253,328 K in an 11.5-dm3 reaction chamber with either CHF2Cl or CH2FCF3 as a reference compound. OH radicals were produced by UV photolysis of an O3,H2O,He mixture at an initial pressure of 200 Torr. Ozone was continuously introduced into the reaction chamber during the UV irradiation. The k1 (298 K) values determined by the absolute method were (1.69 ± 0.07) × 10,15 cm3 molecule,1 s,1 (FP-LIF method) and (1.72 ± 0.07) × 10,15 cm3 molecule,1 s,1 (LP-LIF method), whereas the K1 (298 K) values determined by the relative method were (1.87 ± 0.11) × 10,15 cm3 molecule,1 s,1 (CHF2Cl reference) and (2.12 ± 0.11) × 10,15 cm3 molecule,1 s,1 (CH2FCF3 reference). These data are in agreement with each other within the estimated experimental uncertainties. The Arrhenius rate constant determined from the kinetic data was K1 = (4.71 ± 0.94) × 10,13 exp[,(1630 ± 80)/T] cm3 molecule,1 s,1. Using kinetic data for the reaction of tropospheric CH3CCl3 with OH radicals [k1 (272 K) = 6.0 × 10,15 cm3 molecule,1 s,1, tropospheric lifetime of CH3CCl3 = 6.0 years], we estimated the tropospheric lifetime of CF3CF2CF2CF2CF2CHF2 through reaction with OH radicals to be 31 years. © 2003 Wiley Periodicals, Inc. Int J Chem Kinet 36: 26,33, 2004 [source] Update on peptidylarginine deiminases and deimination in skin physiology and severe human diseasesINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 3 2007M.-C. Méchin Synopsis Deimination (or citrullination) is a recently described post-translational modification, but its consequences are not yet well understood. It is catalysed by peptidylarginine deiminases (PADs). These enzymes transform arginyl residues involved in a peptidyl link into citrullyl residues in a calcium-dependent manner. Several PAD substrates have already been identified like filaggrin and keratins K1 and K10 in the epidermis, trichohyalin in hair follicles, but also ubiquitous proteins like histones. PADs act in a large panel of physiological functions as cellular differentiation or gene regulation. It has been suggested that deimination plays a role in many major diseases such as rheumatoid arthritis, multiple sclerosis, Alzheimer's disease and psoriasis. Five human genes (PADIs), encoding five highly conserved paralogous enzymes (PAD1-4 and 6), have been characterized. These genes are clustered in a single locus, at 1p35-36 in man. Only PAD1-3 are expressed in human epidermis. PADs seem to be controlled at transcriptional, translational and activity levels and they present particular substrate specificities. In this review, we shall discuss these main biochemical, genetic and functional aspects of PADs together with their pathophysiological implications. Résumé La désimination (ou citrullination) est une modification post-traductionnelle catalysée par les peptidyl-arginine désiminases (PADs), décrite depuis peu et dont les conséquences sont encore mal comprises. Ces enzymes transforment, de façon dépendante du calcium, les résidus arginyl engagés dans un lien peptidique en résidus citrullyl. Plusieurs substrats ont été identifiés: la filaggrine et les cytokératines K1 et K10 de l'épiderme, la trichohyaline dans le follicule pileux mais aussi des protéines ubiquistes comme les histones. Les PADs interviennent dans de nombreuses fonctions physiologiques telles que la différenciation cellulaire ou la régulation génique. La désimination pourrait jouer un rôle dans plusieurs maladies sévères et fréquentes comme la polyarthrite rhumatoïde, la sclérose en plaque, la maladie d'Alzheimer ou encore le psoriasis. Cinq gènes humains (PADIs) codant pour 5 enzymes paralogues conservées (PAD1-4 et 6) ont été caractérisés. Ils sont regroupés en un seul locus, en 1p35-36 chez l'homme. Seules les PAD1-3 sont exprimées dans l'épiderme humain. Les PADs semblent contrôlées aux niveaux transcriptionnel et traductionnel, ainsi qu'au niveau de leur activité. Elles présentent chacune leurs spécificités de substrats. Ces principaux aspects biochimiques, génétiques et fonctionnels des PADs tout comme leurs implications physiopathologiques seront discutés dans cette revue. [source] Growth of large protein crystals by a large-scale hanging-drop methodJOURNAL OF APPLIED CRYSTALLOGRAPHY, Issue 4 2010Keisuke Kakinouchi A method for growing large protein crystals is described. In this method, a cut pipette tip is used to hang large-scale droplets (maximum volume 200,µl) consisting of protein and precipitating agents. A crystal grows at the vapor,liquid interface; thereafter the grown crystal can be retrieved by droplet,droplet contact both for repeated macroseeding and for mounting crystals in a capillary. Crystallization experiments with peroxiredoxin of Aeropyrum pernix K1 (thioredoxin peroxidase, ApTPx) and hen egg white lysozyme demonstrated that this large-scale hanging-drop method could produce a large-volume crystal very effectively. A neutron diffraction experiment confirmed that an ApTPx crystal (6.2,mm3) obtained by this method diffracted to beyond 3.5,Å resolution. [source] The effect of body size on food consumption, absorption efficiency, respiration, and ammonia excretion by the inland silverside, Menidia beryllina (Cope) (Osteichthyes: Atherinidae)JOURNAL OF APPLIED ICHTHYOLOGY, Issue 4 2003M. A. Peck Summary The inland silverside, Menidia beryllina (Cope), is an annual zooplanktivore that occurs in estuarine and freshwater habitats along the Atlantic and Gulf of Mexico coasts and drainages of the United States. Experiments were conducted at 25 ± 1°C to quantify the relationship between mean dry weight (WD) and rates of energy gain from food consumption (C), and energy losses as a result of respiration (R) and ammonia excretion (E) during routine activity and feeding by groups of fish. The absorption efficiency of ingested food energy (A) was also quantified. Rates of C, E, and R increased with WD by factors (b in the equation y = aWDb) equal to 0.462, 0.667, and 0.784, respectively. Mean (±SE) rates of energy loss during feeding were 1.6 ± 0.1 (R) and 3.4 ± 0.6 (E) times greater than those for unfed fish. Absorption efficiency was independent of WD and estimated to be 89% of C. From these measurements, the surplus energy available for growth and activity (G) and growth efficiency (K1) were estimated. Over the range in sizes of juveniles and adults (5,500 mg WD), predicted G and K1 values decreased from 7.42 to 0.20 J mg fish,1 day,1 and 63 to 21%, respectively. Measured and predicted bioenergetic parameters are discussed within an ecological context for a northern population of this species. [source] Indirect identification of isoprenoid quinones in Escherichia coli by LC-MS with atmospheric pressure chemical ionization in negative modeJOURNAL OF BASIC MICROBIOLOGY, Issue 6 2004Mengchun Gao Dr. A novel analytical method was applied for identification of isoprenoid quinones in Escherichia coli by liquid chromatography atmospheric press chemical ionization mass spectrometry in negative mode (LC-NI-APCI-MS). Extraction and clean-up of sample were carried out on Sep-Pak Plus Silica solid-phase extraction cartridges. Ubiquinone-7 (UQ-7), Ubiquinone-8 (UQ-8) and Mequinone-8 (MK-8) were determined directly using combined information on retention time, molecular ion mass, fragment ion masses and UV characteristic spectrometry without any standard reagent. It was found that UQ-8 was the major component of isoprenoid quinones in Escherichia coli under aerobic condition. Compared with UQ-8, the relative abundance of UQ-7 and MK-8 is only 15% and 14%, respectively. The average recoveries of UQ-6, UQ-10 and vitamin K1 in Escherichia coli were investigated by standard spiking experiment. The recoveries were achieved in the range from 94 to 106%, and the relative standard deviations (RSD) of the triplicate analysis of the spiked samples (UQ-6, UQ-10 and vitamin K1) ranged from 3 to 8%. The detection limits of LC-NI-APCI-MS were estimated to be 5, 40 and 0.8 ,g/g dry cell for UQ-6, UQ-10 and vitamin K1, respectively. (© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Insulin receptor substrate 1 (IRS-1) plays a unique role in normal epidermal physiology,JOURNAL OF CELLULAR PHYSIOLOGY, Issue 2 2007Marianna Sadagurski Insulin receptor substrate (IRS) proteins play a central role in insulin signaling. Previously we have demonstrated that insulin is essential for normal skin development and function. In the present study we investigated the involvement of the IRS-1 and IRS-2 proteins in skin physiology and in mediating insulin action in skin. For this purpose we have investigated the effects of inactivation of each of the IRSs on skin, studying skin sections and primary skin cells derived from IRS-1 or IRS-2 null mice. We have demonstrated that while the skin of the IRS-2 null mice appeared normal, the skin of the IRS-1 null mice was thinner and translucent. Histological analysis revealed that the thinning of the IRS-1 null skin was a consequence of the thinning of the spinous compartment, consisting of fewer layers. Proliferation of the IRS-1 and IRS-2 null skin epidermal cells was normal. However, the differentiation process of the IRS-1 skin and skin cells was impaired. There was a marked decrease in the induction of the expression of K1, the marker of advanced stages of skin differentiation. In contrary, IRS-2 inactivation had no effects on skin differentiation. In conclusion, we have shown for the first time that IRS-1 but not IRS-2 has an effect on skin formation and development, being one of the main activators of the differentiation process in skin keratinocytes. Furthermore, we suggest that IRS-1 and IRS-2 have distinct roles in skin physiology. J. Cell. Physiol. 213: 519,527, 2007. © 2007 Wiley-Liss, Inc. [source] Differential cytokine expression by human dendritic cells in response to different Porphyromonas gingivalis capsular serotypesJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 10 2009Rolando Vernal Abstract Aim: Capsular polysaccharides play an important role in the virulence of Gram-positive and Gram-negative bacteria. In Porphyromonas gingivalis, six serotypes have been described based on capsular antigenicity and its pathogenicity has been correlated both in vitro and in animal models. This study aimed to investigate the differential response of human dendritic cells (DCs) when stimulated with different P. gingivalis capsular serotypes. Materials and Methods: Using different multiplicity of infection (MOI) of the encapsulated strains K1,K6 and the non-encapsulated K, strain of P. gingivalis, the mRNA expression levels for interleukin (IL)-1,, IL-2, IL-5, IL-6, IL-10, IL-12, IL-13, interferon (IFN)- ,, tumour necrosis factor (TNF)- ,, and TNF- , in stimulated DCs were quantified by real-time reverse transcription-polymerase chain reaction. Results: All P. gingivalis capsular serotypes induced a T-helper type 1 (Th1) pattern of cytokine expression. K1- and K2-stimulated DCs expressed higher levels of IL-1,, IL-6, IL-12p35, IL-12p40, and IFN- , and at lower MOI than DCs stimulated with the other strains. Conclusions: These results demonstrate a differential potential of P. gingivalis capsular serotypes to induce DC responses and a higher capacity of strains K1 W83 and K2 HG184 than other K serotypes to trigger cytokine expression. [source] Purification and Characterization of an ,-L-Rhamnosidase from Aspergillus terreus of Interest in WinemakingJOURNAL OF FOOD SCIENCE, Issue 2 2001M.V. Gallego ABSTRACT: An enzyme with ,-L-rhamnosidase activity was purified to homogeneity from a culture filtrate of Aspergillus terreus after growth in a medium containing L-rhamnose as the sole carbon source. The biosynthesis of this enzyme was repressed by glucose. The enzyme had a molecular mass of 96 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point of 4.6 as determined by analytical isoelectric focusing. The pH and temperature optima for the enzyme were found to be 4.0 and 44 °C, respectively. Using p-nitrophenyl-,-L-rhamnopyranoside as a substrate, the enzyme exhibited Michaelis-Menten kinetics with KM and Vmax values of 0.17 mM and 84 U/mg, respectively. The enzyme was inhibited competitively by L-rhamnose (K1 2.5 mM). Divalent cations such as Ca2+ Mg2+ Zn2+ and Co2+ stimulated the a-L-rhamnosidase activity, whereas this was inhibited by Hg2+ and Cd2+. Ethanol (12% v/v) and glucose (21% w/v) decreased enzyme activity by approximately 20%, while this was not affected by SO2. [source] Perfect coloring and linearly ,-bound P6 -free graphsJOURNAL OF GRAPH THEORY, Issue 4 2007S. A. Choudum Abstract We derive decomposition theorems for P6, K1 + P4 -free graphs, P5, K1 + P4 -free graphs and P5, K1 + C4 -free graphs, and deduce linear ,-binding functions for these classes of graphs (here, Pn (Cn) denotes the path (cycle) on n vertices and K1 + G denotes the graph obtained from G by adding a new vertex and joining it with every vertex of G). Using the same techniques, we also obtain an optimal ,-binding function for P5, C4 -free graphs which is an improvement over that given in [J. L. Fouquet, V. Giakoumakis, F. Maire, and H. Thuillier, 1995, Discrete Math, 146, 33,44.]. © 2006 Wiley Periodicals, Inc. J Graph Theory 54: 293,306, 2007 [source] On totally multicolored starsJOURNAL OF GRAPH THEORY, Issue 3 2006Juan José Montellano-Ballesteros Abstract Given an edge-coloring of a graph G, a subgraph M of G will be called totally multicolored if no two edges of M receive the same color. Let h(G, K1,q) be the minimum integer such that every edge-coloring of G using exactly h(G, K1,q) colors produces at least one totally multicolored copy of K1,q (the q -star) in G. In this article, an upper bound of h(G, K1,q) is presented, as well as some applications of this upper bound. © 2005 Wiley Periodicals, Inc. [source] On possible counterexamples to Negami's planar cover conjectureJOURNAL OF GRAPH THEORY, Issue 3 2004Petr Hlin Abstract A simple graph H is a cover of a graph G if there exists a mapping , from H onto G such that , maps the neighbors of every vertex , in H bijectively to the neighbors of , (,) in G. Negami conjectured in 1986 that a connected graph has a finite planar cover if and only if it embeds in the projective plane. The conjecture is still open. It follows from the results of Archdeacon, Fellows, Negami, and the first author that the conjecture holds as long as the graph K1,2,2,2 has no finite planar cover. However, those results seem to say little about counterexamples if the conjecture was not true. We show that there are, up to obvious constructions, at most 16 possible counterexamples to Negami's conjecture. Moreover, we exhibit a finite list of sets of graphs such that the set of excluded minors for the property of having finite planar cover is one of the sets in our list. © 2004 Wiley Periodicals, Inc. J Graph Theory 46: 183,206, 2004 [source] Spanning 2-trails from degree sum conditionsJOURNAL OF GRAPH THEORY, Issue 4 2004M. N. Ellingham Abstract Suppose G is a simple connected n -vertex graph. Let ,3(G) denote the minimum degree sum of three independent vertices in G (which is , if G has no set of three independent vertices). A 2- trail is a trail that uses every vertex at most twice. Spanning 2-trails generalize hamilton paths and cycles. We prove three main results. First, if ,3G), n - 1, then G has a spanning 2-trail, unless G , K1,3. Second, if ,3(G) , n, then G has either a hamilton path or a closed spanning 2-trail. Third, if G is 2-edge-connected and ,3(G) , n, then G has a closed spanning 2-trail, unless G , K2,3 or K (the 6-vertex graph obtained from K2,3 by subdividing one edge). All three results are sharp. These results are related to the study of connected and 2-edge-connected factors, spanning k -walks, even factors, and supereulerian graphs. In particular, a closed spanning 2-trail may be regarded as a connected (and 2-edge-connected) even [2,4]-factor. © 2004 Wiley Periodicals, Inc. J Graph Theory 45: 298,319, 2004 [source] Defective choosability of graphs with no edge-plus-independent-set minorJOURNAL OF GRAPH THEORY, Issue 1 2004Douglas R. Woodall Abstract It is proved that, if s , 2, a graph that does not have K2,+,Ks,=,K1,+,K1, s as a minor is (s,,1)*-choosable. This completes the proof that such a graph is (s,+,1,,,d,d)*-choosable whenever 0,,,d,,,s,,1 © 2003 Wiley Periodicals, Inc. J Graph Theory 45: 51,56, 2004 [source] Size ramsey numbers of stars versus 4-chromatic graphs,JOURNAL OF GRAPH THEORY, Issue 3 2003Oleg PikhurkoArticle first published online: 31 JAN 200 Abstract We investigate the asymptotics of the size Ramsey number î(K1,nF), where K1,n is the n -star and F is a fixed graph. The author 11 has recently proved that r,(K1,n,F)=(1+o(1))n2 for any F with chromatic number ,(F)=3. Here we show that r,(K1,n,F), n2+o(n2), if , (F) , 4 and conjecture that this is sharp. We prove the case ,(F)=4 of the conjecture, that is, that r,(K1,n,F)=(4+o(1))n2 for any 4-chromatic graph F. Also, some general lower bounds are obtained. © 2003 Wiley Periodicals, Inc. J Graph Theory 42: 220,233, 2003 [source] Another two graphs with no planar coversJOURNAL OF GRAPH THEORY, Issue 4 2001Petr Hlin Abstract A graph H is a cover of a graph G if there exists a mapping , from V(H) onto V(G) such that , maps the neighbors of every vertex , in H bijectively to the neighbors of ,(,) in G. Negami conjectured in 1986 that a connected graph has a finite planar cover if and only if it embeds in the projective plane. It follows from the results of Archdeacon, Fellows, Negami, and the author that the conjecture holds as long as K1,2,2,2 has no finite planar cover. However, this is still an open question, and K1,2,2,2 is not the only minor-minimal graph in doubt. Let ,,4 (,2) denote the graph obtained from K1,2,2,2 by replacing two vertex-disjoint triangles (four edge-disjoint triangles) not incident with the vertex of degree 6 with cubic vertices. We prove that the graphs ,,4 and ,2 have no planar covers. This fact is used in [P. Hlin,ný, R. Thomas, On possible counterexamples to Negami's planar cover conjecture, 1999 (submitted)] to show that there are, up to obvious constructions, at most 16 possible counterexamples to Negami's conjecture. © 2001 John Wiley & Sons, Inc. J Graph Theory 37: 227,242, 2001 [source] Synthesis of (R)- and (S)-[O-methyl - 11C]N -[2-[3-(2-cyano-phenoxy)-2-hydroxy-propylamino]-ethyl]- N,-(4-methoxy-phenyl)-urea as candidate high affinity ,1 -adrenoceptor PET radioligandsJOURNAL OF LABELLED COMPOUNDS AND RADIOPHARMACEUTICALS, Issue 10 2005Stefan Wagner Abstract Molecular imaging and quantification of myocardial ,1 -adrenoceptor (AR) rather than total , -AR density is of great clinical interest since cardiac biopsy studies suggest that myocardial ,1 -AR density is reduced in patients with chronic heart failure whereas cardiac ,2 -AR density may vary. Positron emission tomography (PET), with appropriate radioligands, offers the possibility to assess , -AR density non-invasively in humans. However, no PET radioligand for the selective imaging of cardiac ,1 -ARs is clinically available. Here some derivatives of the well characterized ,1 -AR selective antagonist, ICI 89,406, namely the enantiomers of N -[2-[3-(2-cyano-phenoxy)-2-hydroxy-propylamino]-ethyl]- N,-(4-hydroxy-phenyl)-urea (5a and 5b) were synthesized and evaluated in vitro. The (R)-isomer 5a was more ,1 -selective but has lower affinity than its (S)-enantiomer 5b (,1 -AR selectivity: 6100 vs 1240; ,1 -affinity: K1 = 0.288 nM vs K1 = 0.067 nM). Etherification of the analogous desmethyl precursors, 5e and 5f, respectively, with [11C]iodomethane gave 11C-labelled versions of 5a and 5b, namely 5g and 5h, in 44 ± 5% radiochemical yield (decay-corrected) and 97.4 ± 1.3% radiochemical purity with specific radioactivities of 26.4 ± 9.4 GBq/µmol within 41.2 ± 3.4 min from the end of bombardment (n = 14). 5g and 5h are now being evaluated as candidate radioligands for myocardial ,1 -ARs. Copyright © 2005 John Wiley & Sons, Ltd. [source] Gene expression of differentiation-specific keratins (K4, K13, K1 and K10) in oral non-dysplastic keratoses and lichen planusJOURNAL OF ORAL PATHOLOGY & MEDICINE, Issue 8 2000Balvinder K. Bloor Abstract: Gene expression for the differentiation-specific keratins (K4, K13, K1 and K10) was analyzed in oral non-dysplastic keratoses, oral lichen planus (OLP) and lichenoid reactions (LR) by comparative in situ hybridization (ISH) and immunohistochemistry (IHC) to investigate molecular changes in the altered differentiation pattern from non- to para- or orthokeratinization. At the protein level, K4 and K13 were detected homogeneously in the suprabasal compartment of parakeratotic epithelium but showed reduced expression in orthokeratoses, particularly in the presence of lymphocytes. Corresponding transcripts were restricted to basal and lower prickle cells. Synthesis of K1 and K10 was upregulated and more pronounced in orthokeratotic epithelia. The study showed an alteration in the pattern of differentiation-specific keratins, although involvement of the lymphocytic infiltrate in OLP and LR resulted in further gene modulation. In both diseases, K1 and K10 showed transcriptional control, proteins having the same distribution as their transcripts. This represented a change from post-transcriptional regulation in normal buccal epithelium, in which mRNAs for K1 and K10 are more widely expressed than their proteins. Thus, the pattern of keratin gene expression may be altered in response to frictional/smoking stimuli or immune-mediated mechanisms. [source] Relative in vitro efficacy of the phosphate binders lanthanum carbonate and sevelamer hydrochlorideJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 10 2007Valerie Autissier Abstract The high tablet burden and poor compliance associated with phosphate-binding drugs has led to a search for more potent agents. In vitro -binding studies were performed on the recently introduced binder, lanthanum carbonate (LC; Fosrenol®), to compare its phosphate-binding affinity with sevelamer hydrochloride (SH; RenagelÔ). Langmuir equilibrium binding affinities (K1) for LC and SH were established using different phosphorus (5,100 mM) and binder (134,670 mg per 50 mL) concentrations at pH 3,7, with or without salts of bile acids present (30 mM). At all pH levels, LC had a higher binding affinity for phosphate than SH. For LC, K1 was 6.1,±,1.0 mM,1 and was independent of pH. For SH, K1 was pH dependent, being 1.5,±,0.8 mM,1 at pH 5,7 and 0.025,±,0.002 mM,1 at pH 3, that is, >200 times lower than for LC. In the presence of 30 mM bile salts, SH lost 50% of its phosphate, whereas no displacement of phosphate occurred for LC. These findings indicate that LC binds phosphate more effectively than SH across the pH range encountered in the gastrointestinal tract, and has a lower propensity for bound phosphate to be displaced by competing anions in the intestine. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 96: 2818,2827, 2007 [source] Enantioseparation of nuarimol by affinity electrokinetic chromatography-partial filling technique using human serum albumin as chiral selectorJOURNAL OF SEPARATION SCIENCE, JSS, Issue 18 2008Maria Amparo Martínez-Gómez Abstract The present paper deals with the enantiomeric separation of nuarimol enantiomers by affinity EKC-partial filling technique using HSA as chiral selector. Firstly, a study of nuarimol interactions with HSA by CE-frontal analysis was performed. The binding parameters obtained for the first site of interaction were n1 = 0.84; K1 = 9.7 ± 0.3×103 M,1 and the protein binding percentage of nuarimol at physiological concentration of HSA was 75.2 ± 0.2%. Due to the moderate affinity of nuarimol towards HSA the possibility of using this protein as chiral selector for the separation of nuarimol using the partial filling technique was evaluated. A multivariate optimization approach of the most critical experimental variables in enantioresolution, running pH, HSA concentration and plug length was carried out. Separation of nuarimol enantiomers was obtained under the following selected conditions: electrophoretic buffer composed of 50 mM Tris at pH 7.3; 160 ,M HSA solution applied at 50 mbar for 156 s as chiral selector; nuarimol solutions in the range of 2,8×10,4 M injected hydrodynamically at 30 mbar for 2 s and the electrophoretic runs performed at 30°C applying 15 kV voltage. Resolution, accuracy, reproducibility speed and cost of the proposed method make it suitable for quality control of the enantiomeric composition of nuarimol in formulations and for further toxicological studies. The results showed a different affinity between nuarimol enantiomers towards HSA. [source] PREDICTING SENSORY COHESIVENESS, HARDNESS AND SPRINGINESS OF SOLID FOODS FROM INSTRUMENTAL MEASUREMENTSJOURNAL OF TEXTURE STUDIES, Issue 2 2008R. DI MONACO ABSTRACT The sensory evaluation of cohesiveness, hardness and springiness of 15 solid food samples was performed by eight trained assessors. The rheologic response of the 15 samples was estimated by performing cyclic compression tests and stress,relaxation tests. From the force,deformation curves of the first two cycles of the compression test, texture profile analysis parameters related to cohesiveness, hardness and springiness were calculated. Young's modulus (E), strain (di) and stress (si) at peak as well as irrecoverable strain (ri) and irrecoverable work (Li) were monitored during the first five cycles. From the stress,relaxation response, Peleg's linearization model parameters, K1 and K2, were estimated by best-fit regression. These parameters were used for predicting sensory attributes. Hardness and springiness were both accurately predicted by rheologic properties, while cohesiveness prediction was less representative. PRACTICAL APPLICATIONS This study contributes to enhance the knowledge in the research area of sensory instrumental correlation. Also, the research allows to better understanding that no single instrument is able to measure all texture attributes adequately. In fact, the results demonstrate that both stress,relaxation and cyclic compression tests need to be performed for the correct prediction of sensory responses. [source] Single bolus of intravenous ketamine for anesthetic induction decreases oculocardiac reflex in children undergoing strabismus surgeryACTA ANAESTHESIOLOGICA SCANDINAVICA, Issue 6 2007S. H. Choi Background:, Oculocardiac reflex (OCR) is a major complication of pediatric strabismus surgery. The aim of the present study was to determine whether a single bolus of intravenous (i.v.) ketamine for anesthetic induction can decrease OCR in children undergoing strabismus surgery. Methods:, One hundred and twenty healthy children undergoing strabismus surgery were allocated to three groups using double-blind randomization. Anesthesia was induced with propofol 3 mg/kg in Group P, ketamine 1 mg/kg in Group K1, or ketamine 2 mg/kg in Group K2. Anesthesia was maintained with 3% sevoflurane in 50% N2O/O2 in all patients. The baseline heart rate was obtained 30 s prior to the first traction of the extraocular muscle (EOM). OCR was defined as a development of arrhythmia or a decrease of more than 20% of the baseline heart rate during EOM traction. Results:, The incidence of OCR was significantly lower in the ketamine groups (4/40 and 1/40 in Group K1 and K2, respectively) compared with the propofol group (14/40). Conclusion:, A single bolus of i.v. ketamine 1 or 2 mg/kg for anesthetic induction results in a lower incidence of OCR than propofol when combined with sevoflurane for maintenance in children undergoing strabismus surgery. [source] | ||||||||||||||||||||||||||||||||||||||||