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Selected AbstractsDual enantioselective effect of the insecticide bifenthrin on locomotor behavior and development in embryonic,larval zebrafishENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2010Meiqing Jin Abstract Bifenthrin (BF) is a synthetic pyrethroid that targets the nervous system of insects and may have adverse effects on the behavior and development of nontarget organisms. However, no reports have been issued on the effects of different enantiomers on locomotor behavior for synthetic pyrethroids (SPs) in zebrafish, and whether locomotor activity is associated with the developmental toxicities remains unclear. In this study, enantioselectivity of BF (1S and 1R) on the acute locomotor activity and developmental toxicities of embryonic,larval zebrafish were first evaluated. The results indicated that 1R -BF was more toxic, causing morphological impairments, with a 96-h median effective concentration (EC50) of 226,µg/L for pericardial edema and 145,µg/L for curved body axis. Administration of 20,µg/L of one enantiomer of BF had differential effects on the locomotor activity of zebrafish larvae at 4 d postfertilization (dpf) under alternating light and dark conditions. Larvae treated with 1R -BF were not sensitive to the alteration of light to dark, and the locomotor activities were reduced to a level similar to that observed in light, which otherwise increased rapidly and markedly. However, 1S -BF did not alter the general pattern of zebrafish response to the light or dark compared with the control. The results demonstrated that the differential effects on development might have contributed to the enantioselectivity in the locomotor activity. The consistency of enantioselectivity with insecticidal activity may also indicate a common mode of action. Furthermore, 1R -BF accelerated the spontaneous movement and hatching process, whereas 1S -BF seemed to be inhibitory. The results suggest the need to link behavioral changes to developmental toxicities in order to achieve more comprehensive health risk assessments of chiral pesticides. Environ. Toxicol. Chem. 2010;29:1561,1567. © 2010 SETAC [source] Fluorescence-controlled Er:YAG laser for caries removal in permanent teeth: a randomized clinical trialEUROPEAN JOURNAL OF ORAL SCIENCES, Issue 2 2008Henrik Dommisch The aim of this randomized clinical study was to compare the efficacy of a fluorescence-controlled erbium-loaded yttrium aluminum garnet (Er:YAG) laser with conventional bur treatment for caries therapy in adults. Twenty-six patients with 102 carious lesions were treated using either the Er:YAG laser, at threshold levels of 7, 8, 9, and 10 [U], or rotary burs. Both techniques were applied to each lesion at separate locations. After treatment, dentine samples were obtained using a carbide bur. The viable counts of Streptococcus mutans (SM) and lactobacilli (LB) [expressed as colony-forming units (log10 CFUs)], treatment time, pain, vibration, and sound intensity were determined. The median numbers of CFUs for SM and LB were not statistically different between laser and bur treatment at threshold levels 7 and 8 [U]. At threshold levels 9 and 10 [U], the median number of CFUs for LB [1.11 (range: 0.00,2.04)] were significantly higher following laser treatment than following bur treatment [0.30 (range: 0.00,0.60)]. The results indicate that treatment with a fluorescence-controlled Er:YAG laser at threshold levels of 7 and 8 removed caries to a level similar to that achieved using conventional bur treatment, with clinically irrelevant amounts of remaining bacteria. Although more time consuming, laser treatment provided higher patient comfort than bur treatment. [source] Cell-surface phytase on Pichia pastoris cell wall offers great potential as a feed supplementFEMS MICROBIOLOGY LETTERS, Issue 1 2010Piyanun Harnpicharnchai Abstract Cell-surface expression of phytase allows the enzyme to be expressed and anchored on the cell surface of Pichia pastoris. This avoids tedious downstream processes such as purification and separation involved with extracellular expression. In addition, yeast cells with anchored proteins can be used as a whole-cell biocatalyst with high value added. In this work, the phytase was expressed on the cell surface of P. pastoris with a glycosylphosphatidylinositol anchoring system. The recombinant phytase was shown to be located at the cell surface. The cell-surface phytase exhibited high activity with an optimal temperature at 50,55 °C and two optimal pH peaks of 3 and 5.5. The surface-displayed phytase also exhibited similar pH stability and pepsin resistance to the native and secreted phytase. In vitro digestibility test showed that P. pastoris containing cell-surface phytase released phosphorus from feedstuff at a level similar to secreted phytase. Yeast cells expressing phytase also provide additional nutrients, especially biotin and niacin. Thus, P. pastoris with phytase displayed on its surface has a great potential as a whole-cell supplement to animal feed. [source] Bis(carbazolyl)benzodifuran: A High-Mobility Ambipolar Material for Homojunction Organic Light-Emitting Diode DevicesADVANCED MATERIALS, Issue 37 2009Hayato Tsuji A new ambipolar material bis(carbazolyl)benzodifuran (CZBDF) shows well-balanced and high carrier mobilities for both holes and electrons (>10,3,cm2 V,1 s,1). This new material allows us to fabricate efficient p-i-n homojunction OLEDs that emit light across the full visible color range and perform at a level similar to state-of-the-art heterojunction devices. [source] Induction or suppression of expression of cytochrome C oxidase subunit II by heregulin , 1 in human mammary epithelial cells is dependent on the levels of ErbB2 expressionJOURNAL OF CELLULAR PHYSIOLOGY, Issue 2 2002Yanbo Sun The ErbB family of receptor kinases is composed of four members: epidermal growth factor receptor (EGFR/ErbB1), ErbB2/neu, ErbB3, and ErbB4. Amplification of the ErbB2/neu is found in about 30% of breast cancer patients and is associated with a poor prognosis. Heregulin (HRG) activates the ErbB2 via induction of heterodimerization with ErbB3 and ErbB4 receptors. With suppression subtractive hybridization, we demonstrated that the expression of cytochrome c oxidase subunit II (COXII) is HRG-responsive. Two nontransformed human mammary epithelial cell lines, the HB2 and the HB2ErbB2 (the HB2 engineered to overexpress ErbB2), displayed an opposite response to HRG-mediated regulation. HRG upregulated mRNA expression of COXII in the HB2 cells, but suppressed COXII expression in the HB2ErbB2 cells. A human breast cancer cell line (T47D), which expresses ErbB2 at a level similar to that of the HB2 cells, also responded to HRG by increasing COXII mRNA levels. Therefore, HRG regulation of COXII expression depends on the levels of ErbB2 expression. Furthermore, the expression of COXII was inversely correlated to the levels of ErbB2, i.e., the cells overexpressing ErbB2 exhibited lower COXII levels. HRG-evoked signal transduction differed between the cells with normal ErbB expression and the cells overexpressing ErbB2. The activation of both ERK and PI3-K was essential for HRG regulation of COXII, i.e., blockage of either pathway eliminated HRG-mediated alteration. This is the first report demonstrating that the expression of mitochondria-encoded COXII is HRG-responsive. The levels of ErbB2 expression are decisive for the diverse biological activities of HRG. © 2002 Wiley-Liss, Inc. [source] Increased cysteine availability is essential for cadmium tolerance and accumulation in Arabidopsis thalianaPLANT BIOTECHNOLOGY JOURNAL, Issue 6 2004José R. Domínguez-Solís Summary Employing genetic transformation using an Atcys-3A cDNA construct expressing the cytosolic O -acetylserine(thiol)lyase (OASTL), we obtained two Arabidopsis lines with different capabilities for supplying cysteine under metal stress conditions. Lines 1-2 and 10-10, grown under standard conditions, showed similar levels of cysteine and glutathione (GSH) to those of the wild-type. However, in the presence of cadmium, line 10-10 showed significantly higher levels. The increased thiol content allowed line 10-10 to survive under severe heavy metal stress conditions (up to 400 µm of cadmium in the growth medium), and resulted in an accumulation of cadmium in the leaves to a level similar to that of metal hyperaccumulator plants. Investigation of the epidermal leaf surface clearly showed that most of the cadmium had accumulated in the trichomes. Furthermore, line 10-10 was able to accumulate more cadmium in its trichomes than the wild-type, whereas line 1-2 showed a reduced capacity for cadmium accumulation. Our results suggest that an increased rate of cysteine biosynthesis is responsible for the enhanced cadmium tolerance and accumulation in trichome leaves. Thus, molecular engineering of the cysteine biosynthesis pathway, together with modification of the number of leaf trichomes, may have considerable potential in increasing heavy metal accumulation for phytoremediation purposes. [source] Expression of a putative laccase gene, ZmLAC1, in maize primary roots under stress,PLANT CELL & ENVIRONMENT, Issue 5 2006MINGXIANG LIANG ABSTRACT Laccases are multi-copper-containing glycoproteins and comprise a multi-gene family in plants. However, their physiological functions are still not well understood. We obtained sequence information for a putative laccase gene, ZmLAC1, from maize and studied ZmLAC1 expression in detail. The deduced ZmLAC1 protein was 70% identical to LpLAC5-4, a laccase from ryegrass. ZmLAC1 was expressed in leaves, stems and roots of maize seedlings. In unstressed maize primary roots, a higher ZmLAC1 transcript level was located in the basal region where cell elongation had ceased compared to the apical 5 mm of the roots where cells were rapidly dividing and elongating. A treatment with 300 mm NaCl resulted in a shortened root elongation zone (< 2 mm) and swelling in the apical 5 mm. Associated with the morphological change, the transcript level of ZmLAC1 was enhanced in the apical 5 mm, reaching a level similar to that in the basal region. Other abiotic stresses tested , such as 28.5% polyethylene glycol (PEG), which caused an inhibition of root elongation comparable to 300 mm NaCl , did not affect ZmLAC1 transcript level. Potential roles of ZmLAC1 in the roots responding to NaCl or other high concentration of salts are discussed. [source] The newly synthesized linoleic acid derivative DCP-LA ameliorates memory deficits in animal models treated with amyloid-, peptide and scopolaminePSYCHOGERIATRICS, Issue 4 2005Tetsu NAGATA Abstract Background:, In our earlier study, 8-[2-(2-pentyl-cyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA), a newly synthesized linoleic acid derivative with cyclopropane rings instead of cis -double bonds, facilitated hippocampal synaptic transmission by stimulating glutamate release from presynaptic terminals as mediated via ,7 acetylcholine (Ach) receptors under the influence of protein kinase C. The present study assessed the possibility of using DCP-LA as a cognitive enhancer in animal models. Methods:, Amyloid-,1,40 peptide (300 pM/day) or saline was continuously injected in the right lateral ventricle of rats for 2 weeks. Then, the water maze test was carried out, once per day for 7 days, 1 h after the intraperitoneal injection with DCP-LA or saline. In a different set of experiments, rats were intraperitoneally injected with scopolamine (1 mg/kg) and the water maze test was performed twice per day, with the first test taking place 1 h after the intraperitoneal injection with DCP-LA, galantamine or saline, and the second test starting 2 min after the end of the first. Results:, Continuous intraventricular injection with amyloid-,1,40 peptide in the rat lateral ventricle prolonged the latency for acquisition in the water maze test. DCP-LA (1 mg/kg, intraperitoneal (i.p.)) significantly improved the impairment, which reached a level similar to the latency for sham. Furthermore, DCP-LA (1 mg/kg, i.p.) significantly ameliorated learning and memory deficits in rats treated with scopolamine and was, if not more, effective than galantamine, a modest inhibitor of acetylcholinesterase with nicotinic ACh receptor modulation. Conclusion:, The results of the present study show that DCP-LA ameliorates learning and memory deficits induced by amyloid-,1,40 peptide or scopolamine. DCP-LA may thus offer new hope for dementia patients. [source] Growth, haematological parameters and tissue lipid peroxidation of soft-shelled turtles, Pelodiscus sinensis, fed diets supplemented with different levels of ferrous sulphateAQUACULTURE NUTRITION, Issue 1 2009J.-H. CHU Abstract Soft-shelled turtles, Pelodiscus sinensis, with an average weight of 5.55 g, were fed diets supplemented with eight levels of ferrous sulphate for 8 weeks. The analysed iron content ranged from 50.8 to 482.9 mg kg,1. Growth rate of turtles fed the control diet with no iron supplementation was the lowest among all dietary groups. Haematological parameters including red blood cell, haemoglobin, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration of the turtles fed the control diet were also significantly (P < 0.05) lower relative to the other groups. Thus, dietary iron at 50.8 mg kg,1 (no supplemented iron) was deemed deficient for growth and ineffective at preventing anaemia in juvenile soft-shelled turtle. Whereas, a supplementation of 50 mg kg,1 ferrous sulphate (a total dietary iron of 91.8 mg kg,1) was enough to normalize the haematological values of soft-shelled turtles to the level similar to other iron supplement-fed groups. Within the tested dietary iron range, liver iron content curve-linearly (r2 = 0.99) increased with increasing dietary iron level. Furthermore, thiobarbituric acid-reactive substances in liver tissues of the turtles have also increased when liver iron content increased. The dietary iron requirement of soft-shelled turtle is 120,198 mg kg,1 when ferrous sulphate is used as the source of iron. [source] Host-mediated induction of ,-amylases by larvae of the Mexican bean weevil Zabrotes subfasciatus (Coleoptera: Chrysomelidae: Bruchinae) is irreversible and observed from the initiation of the feeding periodARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 4 2010Thaís D. Bifano Abstract Larvae of Zabrotes subfasciatus secrete , -amylases that are insensitive to the , -amylase inhibitor found in seeds of Phaseolus vulgaris. By analyzing amylase activities during larval development on P. vulgaris, we detected activity of the constitutive amylase and the two inducible amylase isoforms at all stages. When larvae were transferred from the non , -amylase inhibitor containing seeds of Vigna unguiculata to P. vulgaris, the inducible , -amylases were expressed at the same level as in control larvae fed on P. vulgaris. Interestingly, when larvae were transferred from seeds of P. vulgaris to those of V. unguiculata, inducible , -amylases continued to be expressed at a level similar to that found in control larvae fed P. vulgaris continuously. When 10-day-old larvae were removed from seeds of V. unguiculata and transferred into capsules containing flour of P. vulgaris cotyledons, and thus maintained until completing 17 days (age when the larvae stopped feeding), we could detect higher activity of the inducible , -amylases. However, when larvae of the same age were transferred from P. vulgaris into capsules containing flour of V. unguiculata, the inducible , -amylases remained up-regulated. These results suggest that the larvae of Z. subfasciatus have the ability to induce insensitive amylases early in their development. A short period of feeding on P. vulgaris cotyledon flour was sufficient to irreversibly induce the inducible , -amylase isoforms. Incubations of brush border membrane vesicles with the , -amylase inhibitor 1 from P. vulgaris suggest that the inhibitor is recognized by putative receptors found in the midgut microvillar membranes. © 2010 Wiley Periodicals, Inc. [source] Characterization of a Hollow Fiber Bioartificial Liver DeviceARTIFICIAL ORGANS, Issue 5 2005Susan Fugett Abu-Absi Abstract:, A three-compartment bioartificial liver (BAL) has been developed for potential treatment of fulminant hepatic failure. It has been shown previously that viability and liver-specific functions were maintained in laboratory-scale bioreactors of such design. In this study, the performance of hepatocytes in a clinical-scale bioartificial liver was verified by sustained specific production rates of albumin and urea, along with oxygen consumption rates for up to 56 h and liver-specific gene expression for up to 72 h. In addition, transmission of porcine endogenous retrovirus and other type C retroviral particles across the hollow fibers was not detected under both normal and extreme operating fluxes. These results demonstrate that the clinical-scale BAL performs at a level similar to the laboratory scale ,and, that, it, offers, a, viral, barrier, against, porcine , retroviruses. [source] L6-S1 spinal nerve stimulation reduces micturition frequency in anaesthetized rats with cyclophosphamide-induced cystitisBJU INTERNATIONAL, Issue 2 2006FRANÇOIS A. GIULIANO OBJECTIVE To further investigate the rationale for using spinal nerve stimulation (SNS) for treating bladder overactivity associated with cystitis in a rat model of cyclophosphamide-induced cystitis, as several studies suggested that symptoms associated with chronic cystitis could be treated using stimulation of sacral spinal nerves, but the mechanisms by which it works are unknown. MATERIALS AND METHODS Cystitis was induced by i.p. injection of cyclophosphamide 48 h before the experiments in anaesthetized male rats. Neurograms were taken by placing a recording electrode onto the pelvic nerve and a stimulating electrode on either the L6 or S1 ipsilateral spinal nerves. Two selected intensities were then evaluated for SNS in control and cyclophosphamide-treated rats during cystometry. RESULTS Cyclophosphamide resulted in significant bladder overactivity. There was no apparent difference in the neurograms generated in response to SNS of the S1 and L6 spinal nerves, and between cyclophosphamide and control rats. Intensities of 200 µA (A,-fibre-specific) and 2 mA (A,+ C-fibres) were chosen for SNS. Continuous SNS at 200 µA significantly reduced the frequency of voiding and non-voiding contractions in cyclophosphamide-treated rats. SNS at 2 mA resulted in the abolition of voiding contractions, accompanied by continuous leakage of urine. CONCLUSION SNS recruiting only A,-fibre produced fewer voiding contractions in cyclophosphamide-treated rats, to a level similar to that from the control rats. These results support the ability of SNS to decrease bladder overactivity in a pathophysiological model of chemical irritation of the bladder. [source] Altering the Substrate Specificity of RhlI by Directed EvolutionCHEMBIOCHEM, Issue 3 2009Pavan Kumar Reddy Kambam Abstract Reducing virulence: RhlI catalyzes the synthesis of N -butanoyl homoserine lactone (BHL), with a minor product N -hexanoyl homoserine lactone (HHL). By using directed evolution and a genetic screen, RhlI has been engineered for enhanced production of both BHL and HHL at a similar level. Quorum sensing regulates biofilm formation and virulence factor production in the human opportunistic pathogen Pseudomonas aeruginosa. We used directed evolution to engineer RhlI, an enzyme in the RhlI,RhlR quorum-sensing system of P. aeruginosa, to alter its substrate specificity and gain insight into the molecular mechanisms of quorum sensing. By using a genetic screen, we identified a mutant with improved production of RhlI's two signaling molecules, N -butanoyl- and N -hexanoyl-homoserine lactone (BHL and HHL). In particular, production of BHL has been enhanced by more than two-fold, and the synthesis of HHL has been improved from an undetectable level to a level similar to BHL; this change indicates a significant change in substrate specificity. No significant change in the gene expression level was observed. Sequence alignments suggest that the mutations are most likely to facilitate interactions between the enzyme and the two acylated ACP substrates. This work also demonstrates that the genetic screen/selection should be useful in engineering additional quorum-sensing components. [source] The ttgGHI solvent efflux pump operon of Pseudomonas putida DOT-T1E is located on a large self-transmissible plasmidENVIRONMENTAL MICROBIOLOGY, Issue 6 2007José J. Rodríguez-Herva Summary Pseudomonas putida DOT-T1E is a solvent-tolerant strain able to grow in the presence of > 1% (v/v) toluene in the culture medium. A set of multidrug efflux pumps have been found to play a major role in the tolerance of this bacterium to organic solvents (Rojas et al., J Bacteriol 183: 3967,3973). In the course of studies of the mechanisms underlying solvent tolerance in DOT-T1E, we isolated a spontaneous solvent-sensitive mutant derivative which had lost the genes encoding the TtgGHI efflux pump, the most important extrusion element in quantitative terms. Genomic comparisons between the mutant and its parental strain by microarray analysis revealed that in addition to the ttgVW-ttgGHI gene cluster, another group of genes, highly similar to those found in the Tn4653A and ISPpu12 transposable elements of the TOL plasmid pWW0 from P. putida mt-2, were also absent from this strain. Further analysis demonstrated that strain DOT-T1E harboured a large plasmid (named pGRT1) that was lost from the solvent-sensitive mutant. Mapping analysis revealed that the ttgVW-ttgGHI genes and the Tn4653A -like transposon are borne by the pGRT1 plasmid. Plasmid pGRT1 is highly stable and its frequency of loss is below 10,8 per cell per generation under a variety of growth conditions, including nutritional and physical stresses. The pGRT1 plasmid is self-transmissible, and its acquisition by the toluene-sensitive P. putida KT2440 and Pseudomonas aeruginosa PAO1 increased the recipient's tolerance to toluene up to levels similar to those exhibited by P. putida DOT-T1E. We discuss the importance and potential benefits of this plasmid for the development of bacteria with enhanced solvent tolerance, and its potential impact for bioremediation and whole-cell biotransformations. [source] Site-directed mutagenesis of the chemokine receptor CXCR6 suggests a novel paradigm for interactions with the ligand CXCL16EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 8 2008Sarah Abstract Chemokine receptor CXCR6 mediates the chemotaxis and adhesion of leukocytes to soluble and membrane-anchored forms of CXCL16, and is an HIV-1 co-receptor. Here, we describe the effects of mutation of acidic extracellular CXCR6 residues on receptor function. Although most CXCR6 mutants examined were expressed at levels similar to wild-type (WT) CXCR6, an N-terminal E3Q mutant was poorly expressed, which may explain previously reported protective effects of a similar single nucleotide polymorphism, with respect to late-stage HIV-1 infection. In contrast to several other chemokine receptors, mutation of the CXCR6 N,terminus and inhibition of post-translational modifications of this region were without effect on receptor function. Likewise, N-terminal extension of CXCL16 resulted in a protein with decent potency and efficacy in chemotaxis and not, as anticipated, a CXCR6 antagonist. D176N and E274Q CXCR6 mutants were unable to interact with soluble CXCL16, suggesting a critical role for D176 and E274 in ligand binding. Intriguingly, although unable to interact with soluble CXCL16, the E274Q mutant could promote robust adhesion to membrane-anchored CXCL16, suggesting that soluble and membrane-bound forms of CXCL16 possess distinct conformations. Collectively, our data suggest a novel paradigm for the CXCR6:CXCL16 interaction, a finding which may impact the discovery of small-molecule antagonists of CXCR6. [source] The key regulators of adult T helper cell responses, STAT6 and T-bet, are established in early life in miceEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 5 2006Shawn Rose Abstract Murine neonatal immunity is typically Th2 biased. This is characterized by high-level IL-4 production at all phases of the immune response and poor IFN-, memory responses. The differential expression of Th1/Th2 cytokines by neonates and adults could arise if the critical regulators of Th differentiation and function, STAT6 and T-bet, operate differently during the neonatal period. To test this idea, the Th cell responses of wild-type, T-bet-deficient, or STAT6-deficient mice were compared in vitro and in vivo. The absence of these factors had similar qualitative effects on the development of effector function in neonates and adults, i.e., if a Th lineage was inhibited or enhanced in adult animals, a similar phenomenon was observed in neonates. However, there was a striking difference observed in the in vivo Th1 memory responses of STAT6-deficient mice initially immunized as neonates. Antigen-specific IFN-, production was increased 50,100-fold in STAT6-deficient neonates, achieving levels similar to those of STAT6-deficient adults. These findings demonstrate that STAT6 and T-bet signals are central in shaping Th responses in wild-type neonates, as in adult mice, and that the master regulators of Th cell development and function are already firmly established in early life. [source] Notch1 expression on T,cells is not required for CD4+ T,helper differentiationEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 6 2004Fabienne Tacchini-Cottier Abstract Notch1 proteins are involved in binary cell fate decisions. To determine the role of Notch1 in the differentiation of CD4+ Th1 versus Th2 cells, we have compared T,helper polarization in vitro in naive CD4+ T,cells isolated from mice in which the N1 gene is specifically inactivated in all mature T,cells. Following activation, Notch1-deficient CD4+ T,cells transcribed and secreted IFN-, under Th1 conditions and IL-4 under Th2 conditions at levels similar to that of control CD4+ T,cells. These results show that Notch1 is dispensable for the development of Th1 and Th2 phenotypes in vitro. The requirement for Notch1 in Th1 differentiation in vivo was analyzed following inoculation of Leishmania major in mice with a T,cell-specific inactivation of the Notch1 gene. Following infection, these mice controlled parasite growth at the site of infection and healed their lesions. The mice developed a protective Th1 immune response characterized by high levels of IFN-, mRNA and protein and low levels of IL-4 mRNA with no IL-4 protein in their lymph node cells. Taken together, these results indicate that Notch1 is not critically involved in CD4+ T,helper,1 differentiation and in resolution of lesions following infection with L.,major. [source] An MHC anchor-substituted analog of myelin oligodendrocyte glycoprotein,35,55 induces IFN-, and autoantibodies in the absence of experimental autoimmune encephalomyelitis and optic neuritisEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 2 2004Mandy Abstract Previous strategies to ameliorate experimental autoimmune encephalitis (EAE) include the treatment of autoreactive T,cells with altered peptide ligands, which contain amino acid substitutions at TCR contact residues. We recently showed that a variant of myelin oligodendrocyte glycoprotein (MOG),35,55 possessing low affinity for MHC (45D) induced anergy in MOG,35,55-specific T,cells and reduced their encephalitogenicity upon adoptive transfer. Here we investigate the characteristics of the primary immune response to this MHC anchor-substituted peptide. Overall, we observed that immunization with 45D resulted in the production of IFN-, and anti-MOG,35,55 autoantibodies at levels similar to those of MOG,35,55-immunized mice with active EAE. However, no symptoms of clinical or histological EAE or overt histological optic neuritis were observed in 45D-immunized mice. Consistent with this finding, 45D-immunized mice did not exhibit CD4+ infiltrates into the CNS. Therefore, MOG,35,55-specific precursors stimulated with a weak ligand (45D) mediate some EAE-associated effector functions but are unable to fully initiate the inflammatory process in the central nervous system that leads to clinical manifestation of EAE. [source] Amygdala amino acid and monoamine levels in genetically Fast and Slow kindling rat strains during massed amygdala kindling: a microdialysis studyEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 1 2004Rick S. Shin Abstract We investigated the neurochemistry of epileptic seizures in rats selectively bred to be seizure-prone (Fast) vs. seizure-resistant (Slow) to amygdala kindling. Microdialysis was used to measure levels of amino acids [glutamate, aspartate and gamma-aminobutyric acid (GABA)] and monoamines (noradrenaline, dopamine and serotonin) during ,massed' stimulation (MS) (every 6 min) of the ipsilateral amygdala for a total of 40 stimulation trials. Behavioral seizure profiles together with their afterdischarge thresholds (ADTs) and associated durations were assessed during the procedure, and subsequently were redetermined 1, 7 and 14 days later. Then normal ,daily' kindling commenced and continued until the animal reached the fully kindled state. During MS, several generalized seizures were triggered in Fast rats that were associated with long afterdischarge (AD) durations and intermittent periods of elevated thresholds, but in Slow rats, most stimulations were associated with stable ADTs and short ADs. Progressively increasing extracellular glutamate and decreasing GABA was observed in Fast rats during the MS, whereas Slow rats showed levels similar to baseline values. Levels of noradrenaline and dopamine, but not of serotonin, were also increased in both strains throughout the MS treatment. In Fast rats, a dramatic lengthening of AD durations occurred 7 and 14 days following MS, as well as subsequent strong positive transfer to daily kindling, all of which were not seen in Slow rats. Together, these results show that repeated, closely spaced stimulations of the amygdala can differentially alter excitatory and/or inhibitory transmitter levels in a seizure network, and that sensitivity to this manipulation is genetically determined. [source] Local Industry Agglomeration and New Business ActivityGROWTH AND CHANGE, Issue 1 2003Todd Gabe New businesses are highly involved in innovative activity, which enhances worker productivity and leads to increased economic output. This paper investigates the effects of industry concentration on the incidence of new business openings in the 5,504 Maine county-industries. Empirical findings indicate that new business activity increases with the number of incumbent establishments in a county-industry and its concentration level relative to the U.S. economy. Model simulations show that raising county-industries, with no initial industry presence, to concentration levels similar to that of the industry in the U.S. economy results in a 1.7 to 8.9 percent increase in the expected number of business openings over a three-year period. Empirical results also suggest that industry clusters comprised of young and small establishments are more conducive to new business formation than clusters made up of mature and large companies. [source] Comparison of MicroSeal and System B/Obtura II obturation techniquesINTERNATIONAL ENDODONTIC JOURNAL, Issue 12 2003P. R. Cathro Abstract Aim, To compare the proportion of gutta-percha, sealer and voids following the filling of simulated root canals in plastic blocks using two warm gutta-percha techniques. Methodology, Ten simulated root canals in plastic blocks were prepared using System GT Rotary Files® and a .02 taper size 40 Quantec rotary nickel,titanium instrument and then filled with MicroSeal and Kerr EWT sealer. A further 10 simulated canals were prepared solely with System GT Rotary Files® and obturated with System B/Obtura II and Kerr EWT sealer. All blocks were sectioned at 1 mm intervals. The sectioned root fillings were photographed through a stereomicroscope and the resultant slides scanned. The images were analysed using adobe photoshop 6.0 to give the proportions of gutta-percha, sealer or void at each level. The data were analysed using unpaired Student's t -test and Mann,Whitney U -test. Results, The MicroSeal technique produced a dense homogeneous gutta-percha fill at the apical 1 and 2 mm levels similar to the System B/Obtura II technique, but further coronally the sealer became mixed into the MicroFlow gutta-percha producing a heterogeneous mass with significantly less (P < 0.05) solid gutta-percha in the root filling compared to System B/Obtura II. The MicroSeal technique generally had more pooling of cement than the System B technique, however, this was only significantly different (P < 0.05) at the 4 mm level. The System B/Obtura II obturation gave an average of 99.27% ± 1.09 gutta-percha fill at all levels. Conclusions, The MicroSeal technique produced a heterogeneous fill consisting of the MicroFlow Master Cone and gutta-percha from the MicroFlow Cartridge mixed with sealer. The System B/Obtura II technique produced a homogeneous fill at all levels. [source] Patients with Epstein Barr virus-positive lymphomas have decreased CD4+ T-cell responses to the viral nuclear antigen 1INTERNATIONAL JOURNAL OF CANCER, Issue 12 2008Kevin N. Heller Abstract Epstein Barr virus (EBV) causes lymphomas in immune competent and, at increased frequencies, in immune compromised patients. In the presence of an intact immune system, EBV-associated lymphomas express in most cases only 3 or fewer EBV antigens at the protein level, always including the nuclear antigen 1 of EBV (EBNA1). EBNA1 is a prominent target for EBV-specific CD4+ T cell and humoral immune responses in healthy EBV carriers. Here we demonstrate that patients with EBV-associated lymphomas, primarily Hodgkin's lymphoma, lack detectable EBNA1-specific CD4+ T-cell responses and have slightly altered EBNA1-specific antibody titers at diagnosis. In contrast, the majority of EBV-negative lymphoma patients had detectable IFN, expression and proliferation by CD4+ T cells in response to EBNA1, and carry EBNA1-specific immunoglobulins at levels similar to healthy virus carriers. Other EBV antigens, which were not present in the tumors, were recognized in less EBV positive, than negative lymphoma patients, but detectable responses reached similar CD8+ T cell frequencies in both cohorts. Patients with EBV-positive and -negative lymphomas did not differ in T-cell responses in influenza-specific CD4+ T cell proliferation and in antibody titers against tetanus toxoid. These data suggest a selective loss of EBNA1-specific immune control in EBV-associated lymphoma patients, which should be targeted for immunotherapy of these malignancies. © 2008 Wiley-Liss, Inc. [source] Retro-active skill of multi-tiered forecasts of summer rainfall over southern AfricaINTERNATIONAL JOURNAL OF CLIMATOLOGY, Issue 1 2001Willem A. Landman Abstract Sea-surface temperature (SST) variations of the oceans surrounding southern Africa are associated with seasonal rainfall variability, especially during austral summer when the tropical atmospheric circulation is dominant over the region. Because of instabilities in the linear association between summer rainfall over southern Africa and SSTs of the tropical Indian Ocean, the skilful prediction of seasonal rainfall may best be achieved using physically based models. A two-tiered retro-active forecast procedure for the December,February (DJF) season is employed over a 10-year period starting from 1987/1988. Rainfall forecasts are produced for a number of homogeneous regions over part of southern Africa. Categorized (below-normal, near-normal and above-normal) statistical DJF rainfall predictions are made for the region to form the baseline skill level that has to be outscored by more elaborate methods involving general circulation models (GCMs). The GCM used here is the Centre for Ocean,Land,Atmosphere Studies (COLA) T30, with predicted global SST fields as boundary forcing and initial conditions derived from the National Centres for Environmental Prediction (NCEP) reanalysis data. Bias-corrected GCM simulations of circulation and moisture at certain standard pressure levels are downscaled to produce rainfall forecasts at the regional level using the perfect prognosis approach. In the two-tiered forecasting system, SST predictions for the global oceans are made first. SST anomalies of the equatorial Pacific (NIÑO3.4) and Indian oceans are predicted skilfully at 1- and 3-month lead-times using a statistical model. These retro-active SST forecasts are accurate for pre-1990 conditions, but predictability seems to have weakened during the 1990s. Skilful multi-tiered rainfall forecasts are obtained when the amplitudes of large events in the global oceans (such as El Niño and La Niña episodes) are described adequately by the predicted SST fields. GCM simulations using persisted August SST anomalies instead of forecast SSTs produce skill levels similar to those of the baseline for longer lead-times. Given high-skill SST forecasts, the scheme has the potential to provide climate forecasts that outscore the baseline skill level substantially. Copyright © 2001 Royal Meteorological Society [source] The synergistic effects of maltodextrin and high-fructose corn sweetener 90 in a fat-reduced muffinINTERNATIONAL JOURNAL OF CONSUMER STUDIES, Issue 1 2001Frank D. Conforti Abstract Low-fat muffins were prepared using as fat substitutes a combination of maltodextrin and high-fructose corn sweetener 90 (HFCS-90), which replaced sucrose at a rate of 25%, 50%, 75% and 100%. The focus of the investigation was to determine the synergistic effects of these two ingredients on the physical properties in a low-fat muffin. Results indicated that maltodextrin and HFCS-90 were effective in maintaining moisture and water activity (aw) levels similar (P> 0.05) to the full-fat muffin. Texture profiles indicated that the low-fat variations were firmer (P< 0.05) than the full-fat counterpart. Increased levels of HFCS-90 had a significant effect (P< 0.05) on crust and crumb darkening. Maltodextrin alone significantly (P< 0.05) impeded the staling rate during storage. However, when maltodextrin was combined with increased levels of HFCS-90, the staling rate accelerated. The two ingredients were found to be effective in maintaining product quality, but HFCS-90 could only be used up to a 50% replacement before any adverse effects in product quality were observed. [source] Frizzled-1 is involved in the neuroprotective effect of Wnt3a against A, oligomersJOURNAL OF CELLULAR PHYSIOLOGY, Issue 1 2008Marcelo A. Chacón The activation of the canonical Wnt signaling pathway protects hippocampal neurons against the toxicity of Alzheimer's amyloid-,-peptide (A,), however, the role played by the Wnt receptors Frizzleds, has not been studied. We report here that Frizzled-1 mediates the activation of the canonical Wnt/,-catenin pathway by Wnt3a in PC12 cells. In addition, the protective effect of Wnt3a against the toxicity of A, oligomers was modulated by Frizzled-1 expression levels in both PC12 cells and hippocampal neurons. Over-expression of Frizzled-1 significantly increased cell survival induced by Wnt3a and diminished caspase-3 activation, while knocking-down Frizzled-1 expression by antisense oligonucleotides decreased the Wnt3a protection. Over-expression of wild-type ,-catenin, but not a transcriptionally inactive mutated version, prevented the toxicity of A, suggesting that the transcription of Wnt target genes may be involved in these events. This was confirmed by co-transfecting both Frizzled-1 and the inactive form of ,-catenin, which does not elicited protection levels similar to those showed with endogenous ,-catenin. Our results indicate that Wnt3a protects from A,-oligomers toxicity by activating the canonical Wnt signaling pathway through the Frizzled-1 receptor, suggesting a therapeutic potential for this signaling pathway in the treatment of Alzheimer's disease. J. Cell. Physiol. 217: 215,227, 2008. © 2008 Wiley-Liss, Inc. [source] Gingival crevicular fluid laminin-5 ,2-chain levels in periodontal diseaseJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 7 2006Gülnur Emingil Abstract Aim: Our study aimed to examine the molecular forms and gingival crevicular fluid (GCF) levels of laminin-5 ,2-chain in patients with different periodontal disease, and compare the effects of P.gingivalis trypsin-like proteinase on intact laminin-5 ,2-chain species. Methods: Eighteen patients with generalized aggressive periodontitis (G-AgP), 29 patients with chronic periodontitis (CP), 20 with gingivitis and 20 periodontally healthy subjects were included. Probing depth, clinical attachment loss, presence of bleeding on probing and plaque were recorded. Molecular forms and GCF laminin-5 ,2-chain levels and the effects of P. gingivalis trypsin-like proteinase on intact laminin-5 ,2-chain were analysed by computer-quantitated Western immunoblotting. Results: Laminin-5 ,2-chain 40 and 70 kDa fragments could be detected in all groups, in varying levels. The CP group had elevated GCF laminin-5 ,2-chain fragment levels compared with the gingivitis and healthy groups (p<0.008). The G-AgP group had GCF laminin-5 ,2-chain fragment levels similar to the gingivitis and healthy groups (p>0.008). GCF laminin-5 ,2-chain fragments differed clearly from the multiple lower molecular size fragments of P.gingivalis trypsin-laminin-5 ,2-chain proteinases. Conclusion: Increased GCF laminin-5 ,2-chain fragments in periodontitis sites with deep periodontal pocket suggest that these cleaved 40 and 70 kDa fragments could reflect the extent of the inflammatory reaction in CP. [source] Levels of HSP70 in HIV+ patients in different viroimmunological statesJOURNAL OF MEDICAL VIROLOGY, Issue 3 2006E. Espigares Abstract The aim of the study was to establish an association between the viroimmunological status of HIV positive patients and their levels of the 70-kD heat shock protein (Hsp70). The longitudinal retrospective case study involved 61 patients at the University Hospital in Granada (Spain) from 1999 to 2002. Twenty-five patients were viroimmunologically stable, while the other 36 patients had suffered virological failure. A minimum of three blood samples were taken at intervals of at least 3 months for the patients who were stable virologically and immunologically, whereas four samples were taken for the virological failure group: two previous to the onset of virological failure, a third corresponding to the time of virological failure, and a fourth at least 3 months after remission of virological failure. Blood samples were also obtained from 20 healthy control subjects; Hsp70 levels in all were determined by enzyme immunoassay. The mean concentration of Hsp70 was 145.4 ng/ml in the HIV-infected patients as opposed to 72.1 ng/ml in the controls. While the viroimmunologically stable group showed levels similar to those of the controls (66.5 ng/ml), the mean value of Hsp70 in the virological failure group was nearly four times as high (249.1 ng/ml), yet this difference was not statistically significant. The patients treated with reverse transcriptase inhibitors were found to have significantly higher levels of Hsp70 than the other subjects. The transformed variable Hsp70/CD4+ presents less variability than the Hsp70 value itself, giving a higher degree of statistical significance, and may be considered a useful parameter for diagnostic, prognostic, and therapeutic management of HIV positive patients. J. Med. Virol. 78:318,323, 2006. © 2006 Wiley-Liss, Inc. [source] Salivary melatonin response to acute pain stimuliJOURNAL OF PINEAL RESEARCH, Issue 4 2001F.A. Nelson Evidence for a relationship between melatonin, nociception, and analgesia in humans is based on data that are only linked by association and simultaneous occurrence. Studies have reported inverse correlation of the circadian melatonin rhythm with nociception latency and enhancement of opioid analgesia by simultaneous administration of melatonin in animals. This study examines the response of salivary melatonin to acute pain stimuli in 18 healthy subjects ranging in age from 19 to 50 years. A biphasic melatonin response following an acute pain stimulus of 36 V was observed, F(8, 8)=17.839, P<0.001. Within 5 min of the stimulus, melatonin decreased and reached a plateau of 36 pg/mL below baseline by 20 min. This decrease was followed by an increase of 5 pg/mL. Melatonin levels subsequently decreased until they had reached levels similar to those anticipated for the time of day and did not vary thereafter. The magnitude of the melatonin response was not related to age or gender. There was no association between voltage and magnitude of the melatonin responses observed at 15 min (r=0.185, P=0.51) or at 30 min (r=0.468, P=0.09). This study provides the first evidence of melatonin utilization and subsequent pineal gland synthesis following acute pain episodes in humans. [source] The efficacy of computer-based supplementary phonics programs for advancing reading skills in at-risk elementary studentsJOURNAL OF RESEARCH IN READING, Issue 2 2006Paul Macaruso In this study we examined the benefits of computer programs designed to supplement regular reading instruction in an urban public school system. The programs provide systematic exercises for mastering word-attack strategies. Our findings indicate that first graders who participated in the programs made significant reading gains over the school year. Their post-test scores were slightly (but not significantly) greater than the post-test scores of control children who received regular reading instruction without the programs. When analyses were restricted to low-performing children eligible for Title I services, significantly higher post-test scores were obtained by the treatment group compared to the control group. At post-test Title I children in the treatment group performed at levels similar to non-Title I students. [source] Expression of the p16INK4a gene and methylation pattern of CpG sites in the promoter region in rat tumor cell linesMOLECULAR CARCINOGENESIS, Issue 1 2004Kanya Honoki Abstract Loss of p16INK4a protein expression has frequently been related to DNA methylation in association with gene silencing. Although the methylation status of exon1, for p16INK4a involvement in various cancers has been extensively analyzed, it has been pointed out that some inconsistencies existed in its relationship to gene silencing of p16INK4a. In this study, we focused on the expression and methylation status in the regions of nt ,478 to ,201, containing a putative TATA box (nt ,401 to ,396), and nt ,233 to 26, both in a recently cloned 5, upstream region of rat p16INK4a. We showed that rat lung adenocarcinoma RLCNR did not express the p16INK4a gene, whereas rat osteosarcoma COS1NR and malignant fibrous histiocytoma MFH1NR both expressed it at levels similar to normal fibroblasts, even though the region of nt ,233 to 26 was hypermethylated in COS1NR rather than RLCNR. In contrast, the CpG islands near the putative TATA box region were consistently methylated in RLCNR, but not in COS1NR and MFH1NR, as well as in normal fibroblasts. Treatment with 5-aza 2,-deoxycytidine induced expression of p16INK4a gene in RLCNR after 48 h, but no changes were observed in COS1NR and MFH1NR. The results indicated that methylation of CpG islands near a TATA box region played a critical role for gene silencing of the rat p16INK4a gene, rather than that of other regions. © 2003 Wiley-Liss, Inc. [source] | ||||||||||||||||||||||