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Fivefold
Terms modified by Fivefold Selected AbstractsA correlation found between gold concentration in blood and patch test reactions in patients with coronary stentsCONTACT DERMATITIS, Issue 3 2008Susanne Ekqvist Background:, Patients with dental gold restorations are known to have a higher level of gold concentration in blood (B-Au). Objectives:, To further investigate, in a study on patients with intracoronary stents and contact allergy to metals, the gold and nickel release from stainless steel stent with (Au stent) and without (Ni stent) gold plating. Method:, A total of 460 patients treated with stenting underwent patch testing with metals, and information on gold and nickel exposure and blood samples were collected. About 200 blood samples were randomly selected and the analysis of B-Au and nickel concentration in blood (B-Ni) was made using inductively coupled plasma mass spectrometry. Results:, There was a correlation between the intensity of Au patch test reaction and B-Au (P < 0.001). This correlation could not be seen between Ni patch test reaction and B-Ni. A Au stent gave a fivefold higher B-Au than a Ni stent. Conclusions:, Gold is released from the Au stent and patients with a Au stent have a fivefold higher B-Au than patients with an Ni stent. The patch test reactions for gold were correlated with B-Au. [source] Transient expression of serotonin 5-HT4 receptors in the mouse developing thalamocortical projectionsDEVELOPMENTAL NEUROBIOLOGY, Issue 3 2010Erin R. Slaten Abstract The serotonin 5-HT4 receptor (5-HT4 -R) is an unusually complex G-protein coupled receptor that is likely to play important roles in brain development and that may underlie the comorbidity of central and peripheral abnormalities in some developmental disorders. We studied the expression of 5-HT4 -Rs in the developing mouse forebrain at embryonic days 13, 15, 17, and at postnatal days 3 and 14 by using immunohistochemistry, tract tracing, and quantitative RT-PCR. The developing thalamocortical projections transiently expressed 5-HT4 -Rs in the embryonic brain and the 5-HT4 -R expression in the forebrain changed from axonal to somatic around birth. From embryonic days 13,17, the forebrain mRNA levels of the 5-HT4(a) -R and 5-HT4(b) -R splice variants increased nine- and fivefold, respectively, whereas the levels of the 5-HT4(e) -R and 5-HT4(f) -R variants remained relatively low throughout the studied period of embryonic development. These results suggest that during development 5-HT4 -R expression undergoes a dynamic regulation and that this regulation may be important for the normal development of sensory and limbic processing. © 2009 Wiley Periodicals, Inc. Develop Neurobiol, 2010. [source] Effect of chronic denervation and denervation-reinnervation on cytoplasmic creatine kinase transcript accumulationDEVELOPMENTAL NEUROBIOLOGY, Issue 3 2001Charles H. Washabaugh Abstract The extensor digitorum longus (EDL) and soleus muscles of adult mice were chronically denervated or denervated and allowed to reinnervate. Muscles were evaluated 1, 5, 14, 21, and 52 days after sciaticectomy. In terms of weight loss, myofiber atrophy, degeneration, and fibrosis, the soleus muscle was more affected than the EDL by chronic denervation. Fifty-two days after chronic denervation, the number of molecules of MCK/ng total RNA in both muscles (determined with competitive PCR) decreased, with the soleus muscle being more affected. At that stage, BCK mRNA levels in the denervated soleus were unchanged, but they were increased (>50%) in the EDL. Reinnervation restored MCK transcript accumulation in the EDL, whereas, in the soleus MCK, transcripts exceeded control values by 57%, approaching levels in the reinnervated EDL. Despite restoration of MCK mRNA levels, the number of molecules of BCK mRNA/ng total RNA was four- to fivefold higher in reinnervated versus control muscles, suggesting that the genes encoding the CK mRNAs are not coordinately regulated in adult muscle. The role of denervation induced, fiber type changes in regulating CK mRNA accumulation has been evaluated. Electron microscopic analyses have established that fibrosis is not a factor that determines BCK mRNA levels in the chronically denervated or denervated-reinnervated muscles. CK isozyme analyses support the hypothesis that a greater proportion of BCK mRNA found in 52 day chronically denervated and denervated-reinnervated muscles is produced in myofibers vs. nonmuscle cells than in control muscles. © 2001 John Wiley & Sons, Inc. J Neurobiol 47: 194,206, 2001 [source] Age-at-onset and schizophrenia: reversed gender effectACTA PSYCHIATRICA SCANDINAVICA, Issue 4 2002B. N. Gangadhar Objective:,This study seeks an explanation for reversed gender effect on age-at-onset (AAO) in schizophrenia. The hypothesis is older AAO in males would be detected in a sample where higher infant mortality (IMR) prevailed. Method:,Case records of International Classification of Diseases-10 (ICD-10) schizophrenia patients from two states (n=70 each) with an IMR of 13 and 67 per thousand were reviewed and AAO was obtained by using the recorded age and duration of illness. Results:,In the sample from the state with lower IMR, AAO did not differ between the two sexes. However, men had older AAO than women in the state with fivefold higher IMR. Conclusion:,Gender differences in AAO may be a function of perinatal complications. In places where infants with perinatal complications are less likely to survive, hence high IMR, a small group of potentially youngest AAO schizophrenic males may be eliminated thus changing the gender effect on AAO. [source] A dose,response perspective on college drinking and related problemsADDICTION, Issue 2 2010Paul J. Gruenewald ABSTRACT Aims In order to examine the degree to which heavy drinking contributes to risks for problems among college drinkers this paper develops and tests a dose,response model of alcohol use that relates frequencies of drinking specific quantities of alcohol to the incidence of drinking problems. Methods A mathematical model was developed that enabled estimation of dose,response relationships between drinking quantities and drinking problems using self-report data from 8698 college drinkers across 14 campuses in California, USA. The model assumes that drinking risks are a direct monotone function of the amount consumed per day and additive across drinking days. Drinking problems accumulate across drinking occasions and are the basis for cumulative reports of drinking problems reported by college drinkers. Results Statistical analyses using the model showed that drinking problems were related to every drinking level, but increased fivefold at three drinks and more gradually thereafter. Problems were associated most strongly with occasions on which three drinks were consumed, and more than half of all reported problems were related to occasions on which four or fewer drinks were consumed. There were some important differences in dose,responsiveness between men and women and between different groups of ,light', ,moderate' and ,heavier' drinkers. Conclusion Many problems among college students are associated with drinking relatively small amounts of alcohol (two to four drinks). Programs to reduce college drinking problems should emphasize risks associated with low drinking levels. [source] Influence of water chemistry on the acute toxicity of copper and zinc to the cladoceran Ceriodaphnia cf dubiaENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2005Ross V. Hyne Abstract This study determined the influence of key water chemistry parameters (pH, alkalinity, dissolved organic carbon [DOC], and hardness) on the aqueous speciation of copper and zinc and its relationship to the acute toxicity of these metals to the cladoceran Ceriodaphnia cf dubia. Immobilization tests were performed for 48-h in synthetic or natural waters buffered at various pH values from 5.5 to 8.4 (other chemical parameters held constant). The toxicity of copper to C. cf dubia decreased fivefold with increasing pH, whereas the toxicity of zinc increased fivefold with increasing pH. The effect of DOC on copper and zinc toxicity to C. cf dubia was determined using natural fulvic acid in the synthetic water. Increasing DOC was found to decrease linearly the toxicity of copper, with the mean effect concentration of copper that immobilized 50% of the cladocerans (EC50) value 45 times higher at 10 mg/L, relative to 0.1 mg/L DOC at pH 6.5. In contrast, the addition of 10 mg/L DOC only resulted in a very small (1.3-fold) reduction in the toxicity of zinc to C. cf dubia. Copper toxicity to C. cf dubia generally did not vary as a function of hardness, whereas zinc toxicity was reduced by a factor of only two, with an increase in water hardness from 44 to 374 mg CaCO3/L. Increasing bicarbonate alkalinity of synthetic waters (30,125 mg/L as CaCO3) decreased the toxicity of copper up to fivefold, which mainly could be attributed to the formation of copper-carbonate complexes, in addition to a pH effect. The toxicity of copper added to a range of natural waters with varying DOC content, pH, and hardness was consistent with the toxicity predicted using the data obtained from the synthetic waters. [source] Changes in the prevalence of alcohol use in rap song lyrics, 1979,97ADDICTION, Issue 9 2005Denise Herd ABSTRACT Aims This paper explores the role of changing images of drinking and alcoholic beverage use in rap music from its beginnings in the United States in the late 1970s to the late 1990s. Design A sample of 341 rap music song lyrics released from 1979 to 1997 were selected using Billboard and Gavin rating charts. Song lyrics were coded for music genres, alcohol beverage types and brand names, drinking behaviors, drinking contexts, intoxication, attitudes towards alcohol and consequences of drinking. Findings From 1979 to 1997, songs with references to alcohol increased fivefold (from 8 to 44%); those exhibiting positive attitudes rose from 43% to 73%; and brand name mentions increased from 46% to 71%. There were also significant increases in songs mentioning champagne and liquor (mainly expensive brand names) when comparing songs released after 1994 with those from previous years. In addition, there were significant increases in references to alcohol to signify glamour and wealth, and using alcohol with drugs and for recreational purposes. The findings also showed that alcohol use in rap music was much more likely to result in positive than negative consequences. Conclusions Many of these findings are consistent with the idea that rap music has been profoundly affected by commercial forces and the marketing of alcoholic beverages. In addition, it is possible that the increase in references to alcoholic beverages in rap music, particularly spirits, is a reflection of a broader advertising culture which increasingly associates African Americans with alcohol use. [source] Pathologic expression of MHC class,II is driven by mitogen-activated protein kinasesEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2007Isabelle Martins Abstract The class,II transactivator (CIITA) is the master regulator of MHC class,II molecules (MHC,II). In melanoma, the MHC,II are constitutively expressed due to an abnormal transcription of CIITA from its promoter,III (pIII), and requires the presence of a 1-kb enhancer located upstream from this latter. Since mitogen-activated protein kinases (MAPK) have been shown to be activated in most melanomas, we sought to analyze their possible involvement in CIITA expression. Using chemical inhibitors and dominant-negative constructs of MAPK-ERK kinase (Mek1) and MAPK-JNK, we evidenced the inhibition of MHC,II and CIITA expression in melanoma cell lines displaying activated MAPK. Transcriptional regulation by MAPK is known to involve the AP-1 transcription factor family. Sequence analysis revealed an AP-1-responsive motif in the enhancer of CIITA pIII at ,5954/,5947 from the site of transcription initiation. Its mutagenesis reduced CIITA expression four- to fivefold in melanoma cell lines and alleviated the effect of dominant-negative constructs of the MAPK pathway. Together, our findings demonstrate that MAPK-ERK and MAPK-JNK are regulators of CIITA transcription in melanoma, and pinpoint an AP-1-responsive site in the CIITA gene pIII. This should have considerable impact on our understanding of the physio-pathologic expression of MHC,II. [source] Sodium/bicarbonate cotransporter NBCn1/slc4a7 increases cytotoxicity in magnesium depletion in primary cultures of hippocampal neuronsEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 3 2009Deborah S. Cooper Abstract Growing evidence suggests that pharmacological inhibition of Na/H exchange and Na/HCO3 transport provides protection against damage or injury in cardiac ischemia. In this study, we examined the contribution of the sodium/bicarbonate cotransporter NBCn1 (slc4a7) to cytotoxicity in cultured hippocampal neurons of rats. In neurons exposed to extracellular pH (pHo) ranging from 6.2 to 8.3, NBCn1 protein expression increased by fivefold at pH < 6.5 compared to the expression at pHo 7.4. At pHo 6.5, the intracellular pH of neurons was ,1 unit lower than that at pH 7.4. Immunochemistry showed a marked increase in NBCn1 immunofluorescence in plasma membranes and cytosol of the soma as well as in dendrites, at pHo 6.5. NBCn1 expression also increased by 40% in a prolonged Mg2+ -free incubation at normal pHo. Knockdown of NBCn1 in neurons had negligible effect on cell viability. The effect of NBCn1 knockdown on cytotoxicity was then determined by exposing neurons to 0.5 mm glutamate for 10 min and measuring lactate dehydrogenase (LDH) release from neurons. Compared to normal incubation (pHo 7.2 for 6 h) after glutamate exposure, acidic incubation (pHo 6.3 for 6 h) reduced cytotoxicity by 75% for control neurons and 78% for NBCn1-knockdown neurons. Thus, both controls and knockdown neurons showed acidic protection from cytotoxicity. However, in Mg2+ -free incubation after glutamate exposure, NBCn1 knockdown progressively attenuated cytotoxicity. This attenuation was unaffected by acidic preincubation before glutamate exposure. We conclude that NBCn1 has a dynamic upregulation in low pHo and Mg2+ depletion. NBCn1 is not required for acidic protection, but increases cytotoxicity in Mg2+ -free conditions. [source] Identification of amino acids in antiplasmin involved in its noncovalent ,lysine-binding-site'-dependent interaction with plasminFEBS JOURNAL, Issue 9 2003Haiyao Wang The lysine-binding-site-mediated interaction between plasmin and antiplasmin is of great importance for the fast rate of this reaction. It also plays an important part in regulating the fibrinolytic enzyme system. To identify structures important for its noncovalent interaction with plasmin, we constructed seven single-site mutants of antiplasmin by modifying charged amino acids in the C-terminal part of the molecule. All the variants were expressed in the Drosophila S2 cell system, purified, and shown to form stable complexes with plasmin. A kinetic evaluation revealed that two mutants of the C-terminal lysine (K452E or K452T) did not differ significantly from wild-type antiplasmin in their reactions with plasmin, in either the presence or absence of 6-aminohexanoic acid, suggesting that this C-terminal lysine is not important for this reaction. On the other hand, modification of Lys436 to Glu decreased the reaction rate about fivefold compared with wild-type. In addition, in the presence of 6-aminohexanoic acid, only a small decrease in the reaction rate was observed, suggesting that Lys436 is important for the lysine-binding-site-mediated interaction between plasmin and antiplasmin. Results from computerized molecular modelling of the C-terminal 40 amino acids support our experimental data. [source] Coupling of endothelin receptors to the ERK/MAP kinase pathway,FEBS JOURNAL, Issue 20 2001Roles of palmitoylation Endothelins are potent mitogens that stimulate extracellular signal-regulated kinases (ERK/MAP kinases) through their cognate G-protein-coupled receptors, ETA and ETB. To address the role of post-translational ET receptor modifications such as acylation on ERK activation and to identify relevant downstream effectors coupling the ET receptor to the ERK signaling cascades we have constructed a panel of palmitoylation-deficient ET receptor mutants with differential G, protein binding capacity. Endothelin-1 stimulation of wild-type ETA or ETB induced a fivefold to sixfold increase in ERK in COS-7 and CHO cells whereas full-length nonpalmitoylated ETA and ETB mutants failed to stimulate ERK. A truncated ETB lacking the C-terminal tail domain including putative phosphorylation and arrestin binding site(s) but retaining the critical palmitoylation site(s) was still able to fully stimulate ERK activation. Using mutated ET receptors with selective G-protein-coupling we found that endothelin-induced stimulation of G,q, but not of G,i or G,s, is essential for endothelin-mediated ERK activation. Inhibition of protein kinases A and C or epidermal growth factor receptor kinase failed to prevent ETA - and ETB -mediated ERK activation whereas blockage of phospholipase C-, completely abrogated endothelin-promoted ERK activation through ETA and ETB in recombinant COS-7 and native C6 cells. Complex formation of Ca2+ or inhibition of Src family tyrosine kinases prevented ET-1-induced ERK-2 activation in C6-cells. Our results indicate that endothelin-promoted ERK/MAPK activation criticially depends on palmitoylation but not on phosphorylation of ET receptors, and that the G,q/phospholipase C-,/Ca2+/Src signaling cascade is necessary for efficient coupling of ET receptors to the ERK/MAPK pathway. [source] Identification of catalytically important residues in the active site of Escherichia coli transaldolaseFEBS JOURNAL, Issue 8 2001Ulrich Schörken The roles of invariant residues at the active site of transaldolase B from Escherichia coli have been probed by site-directed mutagenesis. The mutant enzymes D17A, N35A, E96A, T156A, and S176A were purified from a talB -deficient host and analyzed with respect to their 3D structure and kinetic behavior. X-ray analysis showed that side chain replacement did not induce unanticipated structural changes in the mutant enzymes. Three mutations, N35A, E96A, and T156A resulted mainly in an effect on apparent kcat, with little changes in apparent Km values for the substrates. Residues N35 and T156 are involved in the positioning of a catalytic water molecule at the active site and the side chain of E96 participates in concert with this water molecule in proton transfer during catalysis. Substitution of Ser176 by alanine resulted in a mutant enzyme with 2.5% residual activity. The apparent Km value for the donor substrate, fructose 6-phosphate, was increased nearly fivefold while the apparent Km value for the acceptor substrate, erythrose 4-phosphate remained unchanged, consistent with a function for S176 in the binding of the C1 hydroxyl group of the donor substrate. The mutant D17A showed a 300-fold decrease in kcat, and a fivefold increase in the apparent Km value for the acceptor substrate erythrose 4-phosphate, suggesting a role of this residue in carbon,carbon bond cleavage and stabilization of the carbanion/enamine intermediate. [source] Human Werner helicase interacting protein 1 (WRNIP1) functions as a novel modulator for DNA polymerase ,GENES TO CELLS, Issue 1 2005Toshiki Tsurimoto Human WRNIP1, a Werner DNA helicase interacting protein 1, was expressed in insect cells and E. coli. The purified protein behaved as a homo-oligomeric complex with a native molecular mass indicative of an octamer, and the complex copurified with an ATPase activity that was stimulated by double-stranded DNA ends. As suggested by genetic studies of budding yeast WRNIP1/Mgs1, the purified human WRNIP1 complex interacted physically with human DNA polymerase , (pol ,), stimulating its DNA synthesis activity more than fivefold in the presence or absence of proliferating cell nuclear antigen. Analysis of reaction products demonstrated the stimulation to be partly due to an increased processivity of pol , but more importantly to an increase in its initiation frequency. Addition of ATP to reactions partially suppressed stimulation by WRNIP1. Furthermore, a mutant WRNIP1 lacking ATPase activity could stimulate pol , normally but was insensitive to suppression by ATP. These results indicate that WRNIP1 functions as a modulator for initiation or restart events during pol ,-mediated DNA synthesis and that its ATPase activity is utilized to sense DNA ends and to regulate the extent of stimulation. [source] Causes of the first AIDS-defining illness and subsequent survival before and after the advent of combined antiretroviral therapy,HIV MEDICINE, Issue 4 2008S Grabar Objectives To analyse the impact of combined antiretroviral treatment (cART) on survival with AIDS, according to the nature of the first AIDS-defining clinical illness (ADI); to examine trends in AIDS-defining causes (ADC) and non-AIDS-defining causes (non-ADC) of death. Methods From the French Hospital Database on HIV, we studied trends in the nature of the first ADI and subsequent survival in France during three calendar periods: the pre-cART period (1993,1995; 8027 patients), the early cART period (1998,2000; 3504 patients) and the late cART period (2001,2003; 2936 patients). Results The three most frequent initial ADIs were Pneumocystis carinii (jirovecii) pneumonia (PCP) (15.6%), oesophageal candidiasis (14.3%) and Kaposi's sarcoma (13.9%) in the pre-cART period. In the late cART period, the most frequent ADIs were tuberculosis (22.7%), PCP (19.1%) and oesophageal candidiasis (16.2%). The risk of death after a first ADI fell significantly after the arrival of cART. Lower declines were observed for progressive multifocal leukoencephalopathy, lymphoma and Mycobacterium avium complex infection. After an ADI, the 3-year risk of death from an ADC fell fivefold between the pre-cART and late cART periods (39%vs. 8%), and fell twofold for non-ADCs (17%vs. 9%). Conclusions The relative frequencies of initial ADI have changed since the advent of cART. Tuberculosis is now the most frequent initial ADI in France; this is probably the result of the increasing proportion of migrants from sub-Saharan Africa. After a first ADI, cART has a major impact on ADCs and a smaller impact on deaths from other causes. The risk of death from AIDS and from other causes is now similar. [source] Influence of the surface on thrombin generationINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 4 2008T. W. STIEF Summary Thrombin generation depends on the surface of the blood vessel or container. With a new ultra-sensitive and -specific thrombin assay the surface-dependent thrombin generation was quantified. Citrated blood or plasma was preincubated for 1 h (37 °C). Citrated blood, plasma, or plasma with 0,10 g/l hemoglobin,erythrocyte microparticles (Hb,MP) were preincubated at 23 °C or at 37 °C. Plasma samples (50 ,l) were recalcified in polystyrol (PS) wells and incubated for different coagulation reaction times (CRT). Final supramolar arginine concentrations, 0.1% Triton X 100, and chromogenic thrombin substrate concentrations in the onefold km,range were added and the linear ,A/t was measured in the recalcified coagulation activity assay (RECA). Aprotinin or corn trypsin inhibitor were added. (i) Recalcification of plasma (in different monovettes) pre-incubated for 1 h (37 °C) generated the following thrombin activities after 7 min (37 °C): 0.74 IU/ml (polypropylene (PP)-citrate), 0.39 IU/ml (PP-EDTA), 0.06 IU/ml (PP-heparin), 1.38 IU/ml (PS), 0.63 IU/ml (1 ml volume PP), 0.13 IU/ml (15 ml volume PP), and 3.62 IU/ml (glass). (ii) Recalcification of preincubated whole blood generated up to about fivefold more thrombin. (iii) Thrombin generation is proportional to the plasmatic concentration of Hb,MP, 10 g/l Hb,MP generating about 4 IU/ml thrombin within 20 min CRT. (iv) The IC50 of aprotinin and corn typsin inhibitor on thrombin generation in RECA are about 2 KIU/ml and about 1 U/ml, respectively. The reaction wall, the preincubation temperature, and hemolysis influences thrombin generation. The RECA allows to diagnose the prothrombotic capacity of any material. [source] Pattern of cross-resistance in pyrethroid-selected populations of Helicoverpa armigera Hübner (Lep., Noctuidae) from IndiaJOURNAL OF APPLIED ENTOMOLOGY, Issue 9-10 2004T. Ramasubramanian Abstract:, In Helicoverpa armigera, withdrawal of selection pressure resulted in a two- to fourfold increase in susceptibility to synthetic pyrethroids and continuous selection enhanced the resistance level by four- to fivefold to the respective pyrethroids at the end of the 14th generation. Populations selected for resistance to one pyrethroid showed positive cross-resistance to all other pyrethroids, but no cross-resistance to endosulfan and thiodicarb. There was a significant increase in mixed-function oxidase activity with advancing generation suggesting its possible role in the positive cross-resistance among the pyrethroids. The induction of carboxyl esterases in pyrethroid-selected populations may have resulted in the activation of indoxacarb, thereby accounting for the observed negative cross-resistance. [source] Effect of a transient perturbation on marine bacterial communities with contrasting historyJOURNAL OF APPLIED MICROBIOLOGY, Issue 3 2010O. Zemb Abstract Aims:, To evaluate the importance of the bacterial composition on the resilience of the organic matter assimilation in the sea. Methods and Results:, Chemostats were inoculated with coastal and offshore bacterial communities. Bacterial density and protein synthesis increased before stabilizing, and this response to confinement was more marked in the offshore chemostats. Before the toluene perturbation the community structure in the coastal chemostats remained complex whereas the offshore chemostats became dominated by Alteromonas sp. After the perturbation, bacterial protein synthesis was inhibited before peaking briefly at a level fivefold to that observed before the perturbation and then stabilizing at a level comparable to that before the perturbation. Alteromonas dominated both the coastal and the offshore communities immediately after the perturbation and the coastal communities did not recover their initial complexity. Conclusions:, Cell lysis induced by the toluene perturbation favoured the growth of Alteromonas which could initiate growth rapidly in response to the nutrient pulse. Despite their different community structure in situ, the resilience of protein synthesis of coastal and offshore bacterial communities was dependent on Alteromonas, which dominated in the chemostats. Significance and Impact of the Study:, Here we show that although Alteromonas sp. dominated in artificial offshore and coastal communities in chemostats, their response time to the shock was different. This suggests that future perturbation studies on resilience in the marine environment should take account of ecosystem history. [source] Effect of recombinant Rv1009 protein on promoting the growth of Mycobacterium tuberculosisJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2008X. Wu Abstract Aims:, To determine whether resuscitation-promoting factor (RPF) from Mycobacterium tuberculosis can promote mycobacterial growth and shorten culture time. Method and Results:, We cloned, expressed and purified an RPF from M. tuberculosis, Rv1009 protein and subsequently studied the biological activity of the recombinant Rv1009 (rRv1009) in liquid and on solid media. Our results indicate that the molecular weight of rRv1009 protein expressed in Escherichia coli BL21 was approximately 39 kDa. At picomolar and micromolar concentrations, rRv1009 protein could increase the optical density of freeze-dried Mycobacterium bovis BCG three to fivefold in Middlebrook 7H9 medium, stimulate the growth of viable mycobacteria on solid medium, and shorten positive growth detection time of a small number of M. tuberculosis in BACTEC 960 medium. Conclusions:, The rRv1009 could promote proliferation of mycobacteria. It may be useful for culture of mycobacteria presented in clinical samples. Significance and Impact of the Study:, rRv1009 protein can be used as a growth-promoting reagent of mycobacteria in the medium to shorten the time of culture. [source] Allergenicity testing of supermethrin, phenoxyacetic acid and DNCB using in vivo and in vitro modifications of the local lymph node assays, maximization and epicutaneous testingJOURNAL OF APPLIED TOXICOLOGY, Issue 4 2001M. Kuricova Abstract The purpose of this study was to compare two methods of testing for allergenicity: in vivo and in vitro modifications of local lymph node assays (LLNA) in mice and the maximization and epicutaneous skin tests in guinea pigs as per the Organization for Economic Cooperation and Development (1981). Two pesticides,the synthetic pyrethroid insecticide supermethrin (SM) and the herbicide phenoxyacetic acid (PAA),were evaluated using this testing battery. 1-Chloro-2,4-dinitrobenzene (DNCB) was selected as a reference allergen for the local lymph node assay. In vitro modification of LLNA proliferative response per standard cell count in lymphocyte cultures derived from treated Balb/c mice did not differ from control mice. Results of the in vivo modification showed that treatment with 50% PAA and 50% SM resulted in a lower proliferation response of lymphocytes in lymph nodes compared with control animals. The vigour of the proliferative response varied more in in vivo modification of LLNA. Stimulation indices were <3, so PAA and SM did not indicate classification as allergens. Lymphocyte proliferation in 1% DNCB-activated lymph nodes was approximately fivefold higher than in those derived from control mice. Proliferation response in vitro calculated as stimulation index was higher in DNCB-treated mice than those observed in vivo, but differences were not dramatic. Auricular lymph node weight and cellularity in mice treated with PAA and SM were similar to controls. The DNCB stimulation index for lymph node cellularity was 5.5. Lymph node weight was three times higher in comparison with controls. In the maximization test in guinea pigs SM and PAA acid resulted in 40% and 50% of animals demonstrating sensitization, respectively. Epicutaneous administration resulted in weaker reaction. Both SM and PAA are mildly strong sensitizers by this battery. Copyright © 2001 John Wiley & Sons, Ltd. [source] The characterization and optimization of injectable silicone resin particles in conjunction with dermal fibroblasts and growth factors: An in vitro studyJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2010Robert M. Crews Abstract Minimally invasive subdermal injection of liquid silicone has been used clinically to augment the soft tissue of the foot to mitigate high pressures that cause diabetic foot ulcers. However, implant migration has been a clinical issue. The objective of this study was to assess the effects of three specific concentrations of silicone resin particles (12 ,m average diameter) in conjunction with either platelet-derived growth factor (PDGF-BB) or basic fibroblast growth factor (bFGF) on fibroblast cell proliferation, collagen synthesis, cell morphology, and migration through in vitro assays and a monolayer scratch wound model. PDGF and bFGF enhanced the proliferation of fibroblasts 5.7-fold and fivefold, respectively, while the addition of silicone particles had no significant effect on proliferation. Collagen production was increased approximately twofold with the addition of bFGF and the medium concentration of particles over bFGF without particles and the PDGF groups. The addition of silicone particles had no significant effect on collagen production compared with control groups without particles. Fibroblast migration was enhanced by the addition of both PDGF and bFGF compared to controls, although slower scratch wound closure rates were observed in the presence of particles compared to controls without particles. Cell morphology suggested that particles induced cellular aggregation encircling silicone particles postwounding as well as migration into the wound area. These results suggest that silicone particles in combination with a growth factor might enhance fibroblast aggregation and implant stability, and could promote connective tissue ingrowth and implant encapsulation in the soft tissue of the diabetic foot. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010 [source] Competitive effects of grasses and woody plants in mixed-grass prairieJOURNAL OF ECOLOGY, Issue 4 2001Duane A. Peltzer Summary 1,Variation in the competitive ability of plant species may determine their persistence and abundance in communities. We quantified the competitive effects of grasses and woody plants in native mixed-grass prairie on the performance of transplant species and on resources. 2,We separated the effects of grasses, shrubs and intact vegetation containing both grasses and shrubs by manipulating the natural vegetation using selective herbicides to create four neighbourhood treatments: no neighbours (NN), no shrubs (NS), no grasses (NG) and all neighbours (AN). Treatments were applied to 2 × 2 m experimental plots located in either grass- or shrub-dominated habitats. The effects of grasses and shrubs on resource availability (light, soil moisture, soil available nitrogen) and on the growth of transplants of Bouteloua gracilis, a perennial tussock grass, and Elaeagnus commutata, a common shrub, were measured over two growing seasons. 3,Resource availability was two- to fivefold higher in no neighbour (NN) plots than in vegetated plots (NS, NG, AN) with grasses and shrubs having similar effects. Light penetration declined linearly with increasing grass or shrub biomass, to a minimum of about 30% incident light at 500 g m,2 shoot mass. Soil resources did not decline with increasing neighbour shoot or root mass for either grasses or shrubs, suggesting that the presence of neighbours was more important than their abundance. 4,Transplant growth was significantly suppressed by the presence of neighbours, but not by increasing neighbour shoot or root biomass, except for a linear decline in Bouteloua growth with increasing neighbour shoot mass in plots containing only shrubs. Competition intensity, calculated as the reduction in transplant growth by neighbours, was similar in both grass- and shrub-dominated habitats for transplants of Bouteloua, but was less intense in shrub-dominated habitats for the shrub Elaeagnus. Variation in the persistence and abundance of plants in communities may therefore be more strongly controlled by variation in the competitive effects exerted by neighbours than by differences in competitive response ability. [source] RECOVERY OF SINAPIC ACID FROM A WASTE STREAM IN THE PROCESSING OF YELLOW MUSTARD PROTEIN ISOLATEJOURNAL OF FOOD PROCESS ENGINEERING, Issue 2 2008N. PRAPAKORNWIRIYA ABSTRACT A large amount of waste permeates generated from the processing of yellow mustard protein was concentrated fivefold using a nanofilter with a molecular weight cut off of 1,000 Da, while approximately 74% of sinapic acid was retained. Sinapic acid was then released from sinapine, its esterified form, by an alkaline hydrolysis. The hydrolyzed solution was acidified to prevent oxidation of the sinapic acid and to precipitate the remaining proteins. Subsequently, sinapic acid and other phenolics were extracted by a two-stage extraction using a mixture of diethyl ether and ethyl acetate (1:1), 165-min extraction time and permeate-to-solvent ratio of 1:2. Approximately 95% of the sinapic acid in the acidified permeate was finally concentrated in the solvent phase. PRACTICAL APPLICATIONS This development has led to an economical process to recover phenolics and to treat effluent from a process of oilseed protein while reducing the use of water during the extraction of protein. A reduction of water usage makes the processing of oilseed protein isolate more economically attractive, and the recovered phenolics may find a use as a nutraceutical. The developed process is not only limited to the recovery of phenolics from mustard, but also applied for recovering phenolic acids, specifically sinapic acid, from waste water from membrane processing of protein from mustard and similar polyphenol-containing oilseeds. [source] In vivo pharmacology and antidiarrheal efficacy of a thiazolidinone CFTR inhibitor in rodentsJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 1 2005N.D. Sonawane Abstract A small-molecule inhibitor of the cystic fibrosis transmembrane conductance regulator (CFTR), 3-[(3-trifluoromethyl)phenyl]-5-[(4-carboxyphenyl)methylene]-2-thioxo-4-thiazolidinone (CFTRinh -172), reduces enterotoxin-induced intestinal fluid secretion in rodents. Here, we study CFTRinh -172 pharmacology and antidiarrheal efficacy in rodents using 14C-labeled CFTRinh -172, liquid chromatography/mass spectrometry, and a closed intestinal loop model of fluid secretion. CFTRinh -172 was cleared primarily by renal glomerular filtration without chemical modification. CFTRinh -172 accumulated in liver within 5 min after intravenous infusion in mice, and was concentrated fivefold in bile over blood. At 30,240 min, CFTRinh -172 was found mainly in liver, intestine, and kidney, with little detectable in the brain, heart, skeletal muscle, or lung. Pharmacokinetic analysis in rats following intravenous bolus infusion showed a distribution volume of 770 mL with redistribution and elimination half-times of 0.14 h and 10.3 h, respectively. CFTRinh -172 was stable in hepatic microsomes. Closed-loop studies in mice indicated that a single intraperitoneal injection of 20 ,g CFTRinh -172 inhibited fluid accumulation at 6 h after cholera toxin by >90% in duodenum and jejunum, ,60% in ileum and <10% in colon. No toxicity was seen after high-dose CFTRinh -172 administration (3 mg/kg/day in two daily doses) in mice over the first 6 weeks of life. The metabolic stability, enterohepatic recirculation, slow renal elimination, and intestinal accumulation of CFTRinh -172 account for its efficacy as an antidiarrheal. © 2004 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 94:134,143, 2005 [source] Nurse staffing, bed numbers and the cost of acute psychiatric inpatient care in EnglandJOURNAL OF PSYCHIATRIC & MENTAL HEALTH NURSING, Issue 8 2008L. BOWERS rmn phd The aim of this analysis was to describe the composition, variability and factors associated with nurse staffing costs in acute psychiatric inpatient care. Numbers of acute inpatient beds in England have fallen, creating an occupancy crisis. Numbers of acute inpatient nursing staff are linked to quality of care. Variance in staffing and beds has considerable resource implications, but little is known about how these costs are structured. The sample comprised survey data from 136 wards in 26 NHS Trusts, matched with nationally available data on service levels, population and outcomes. The cost of providing acute inpatient care varied fivefold between different Trusts. This variation comprised of numbers of beds/population, numbers of nurses/beds and the proportion of nurses qualified. These variations were not fully accounted for by differing levels of social deprivation. Although service provision levels in London were higher, wide variation in costs existed in every region. Associations between nursing cost per bed and performance indicators were found. As investment in acute inpatient care varies widely, we need to know much more about the relationship of inputs to outputs, so that empirically based standard service levels can be defined. [source] Differential effects of lorazepam on sleep and activity in C57BL/6J and BALB/cJ strain miceJOURNAL OF SLEEP RESEARCH, Issue 3 2009XIANGDONG TANG Summary Compared to C57BL/6 mice, BALB/c mice exhibit greater ,anxiousness' on behavioural tests of anxiety, and can show significantly longer sleep disruptions after exposure to anxiogenic situations. Relative to C57BL/6 mice, BALB/c mice also have reduced benzodiazepine (BZ) receptor densities in the brain and fivefold less BZ receptor density in the amygdala, a region important in anxiety and in the control of arousal. Lorazepam is a BZ receptor full agonist and has been used to treat both anxiety and insomnia. Differences between C57BL/6 and BALB/c mice raise the question of whether BZ agonists would impact sleep and activity differentially in the two strains. We examined the effects of two doses of lorazepam (0.5 and 1.5 mg kg,1) or saline alone (0.2 mL) on sleep and activity in C57BL/6 (n = 8) and BALB/c (n = 7) mice. Compared to saline, both doses of lorazepam significantly increased non-rapid eye movement (NREM) and reduced activity in both strains. In C57BL/6 mice, rapid eye movement (REM) was increased at both doses. In BALB/c mice, the 0.5 mg kg,1 dose had no significant influence on REM, whereas REM was reduced significantly after the 1.5 mg kg,1 dose. The results demonstrate significant differences between C57BL/6 and BALB/c mice in the effects of lorazepam on REM, whereas the effects on NREM and activity were similar. Strain differences in the number of BZ receptors in the amygdala, but not other brain regions, suggests possible site specificity in the effects of lorazepam on REM. These differences in BZ-binding sites in the amygdala could be a significant factor in differences in the sleep response between C57 and BALB/c mice. [source] Contents of carotenoids, ascorbic acid, minerals and total glucosinolates in leafy brassica pakchoi (Brassica rapa L. chinensis) as affected by season and varietyJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 5 2009Peter Hanson Abstract BACKGROUND: Pakchoi (Brassica rapa L. chinensis) is economically and nutritionally important in East and Southeast Asia. The objective of this study was to evaluate the effects of season and variety on its phytonutrient content (carotenoids, minerals, ascorbic acid and total glucosinolates). RESULTS: Thirty-five varieties were evaluated in one dry season and one wet season field trial at AVRDC,The World Vegetable Center, Taiwan. Mean carotenoid content was about 36% greater in the dry season than in the wet season trial. In contrast, ascorbic acid and total glucosinolate contents were 48 and 72% higher respectively in the wet season than in the dry season trial. Dry matter, calcium and iron contents were also 69, 69 and 21% greater respectively in the wet season than in the dry season trial. Significant differences among entries were found for most phytonutrients; the ranges of variety means were fourfold for total carotenoids, fivefold for iron and total glucosinolates and twofold for calcium. CONCLUSION: Season strongly influences the phytonutrient content of pakchoi grown in the tropics. Significant differences among entries were found for all phytonutrients, and there is potential to increase levels of individual phytonutrients through plant breeding. However, breeding would need to target varieties for either the dry or the wet season. Copyright © 2009 Society of Chemical Industry [source] Treatment of chronic hepatitis C virus infection in patients with cirrhosisJOURNAL OF VIRAL HEPATITIS, Issue 5 2000Zeuzem Chronic hepatitis C virus (HCV) infection eventually leads to cirrhosis in 20,30% of patients and to hepatocellular carcinoma (HCC) in 1,5% of patients. Rates of sustained virological response with standard interferon-, (IFN-,) are low in patients without cirrhosis (generally < 20%) and are even lower in those with cirrhosis. Combination therapy with IFN and ribavirin improves response rates in patients with chronic hepatitis C without cirrhosis, and the results from subgroups of HCV-infected patients with advanced fibrosis or cirrhosis are encouraging. Importantly, treatment with IFN slows progression of liver fibrosis, regardless of HCV genotype or early response to therapy, and reduces the risk of HCC by two- to fivefold. The risk of development of HCC is also lower in patients who show at least a partial response to IFN therapy compared with those who show no response. There is a clear need for more definitive studies of treatment in patients with chronic hepatitis C and cirrhosis, ideally using therapies with greater efficacy. Nonetheless, based on the potential to slow the progression of liver fibrosis (regardless of treatment response) and to reduce the risk of HCC, a greater number of HCV-infected patients with cirrhosis should be considered as candidates for IFN treatment. Preliminary data indicate that pegylated IFNs have improved virological response rates and may have additional clinical benefits in the prevention or reduction of fibrosis and retardation of progression of cirrhosis and HCC in these patients. [source] Rangeland development of the Mu Us Sandy Land in semiarid China: an analysis using Landsat and NOAA remote sensing dataLAND DEGRADATION AND DEVELOPMENT, Issue 2 2003M. C. Runnström Abstract Degradation of the dry semiarid ecosystems in the Mu Us Sandy Land of north central China was explored using high-resolution satellite images from 1978, 1987 and 1996. This study monitored both changes in grassland biomass production and reclamation activities to detect the nature and scale of land degradation since major economic reforms were introduced in 1978. The position of the high-resolution images within the vegetation cycles was inspected from National Oceanographic and Atmospheric Administration (NOAA) NDVI images at 10-day repetition and seasonal precipitation patterns. A model was developed to categorize changes in the vegetation signal activity from 30,×,30,m pixels into vegetation cover development and land-use changes between 1987 and 1996. A general increase of biomass production was evident despite the rapid increase in numbers of grazing animals. This increase in biomass was confirmed by the NOAA time series, which also revealed annual variability related to the amount and pattern of the seasonal rains. Rangeland conversion to farmland was detected, and this process has increased the area of cultivation almost fivefold. The classified area of cultivation corresponds with reported statistical records, also showing that irrigation features in virtually 100 per cent of the sown area. Signs of declining biological production, indicating land degradation processes, are few. Biomass production has increased, with a gain in the economic output from both crop and animal production. The early start of active measures to halt desertification has increased vegetation cover and lowered wind erosion potential and grasslands seems to be managing the high levels of grazing pressure. Copyright © 2003 John Wiley & Sons, Ltd. [source] Colorectal cancer prevention in ulcerative colitis: a case-control studyALIMENTARY PHARMACOLOGY & THERAPEUTICS, Issue 2 2000Eaden Background: The risk of colorectal cancer (CRC) in ulcerative colitis (UC) increases with extent and duration of disease. Identifying other risk factors would allow targeting of sub-groups at greatest risk, enabling more cost-effective surveillance. Methods: We conducted a case-control study comparing 102 cases of CRC in UC with matched controls. Odds ratios (OR) for cancer risk were estimated by conditional logistic regression. A multivariate model assessed the contribution of individual variables. Results: Regular 5-aminosalicylic acid (5-ASA) therapy reduces cancer risk by 75% (OR 0.25, 95% CI: 0.13,0.48, P < 0.00001). Adjusting for other variables, taking mesalazine regularly reduces risk by 81% (OR 0.19, 95% CI: 0.06,0.61, P=0.006) and visiting a hospital doctor more than twice a year also reduces risk (OR 0.16, 95% CI: 0.04,0.60, P=0.007). Considering variables independently, having a family history of sporadic CRC in any relative increases risk fivefold (OR 5.0, 95% CI: 1.10,22.82, P < 0.04). Conclusions: CRC risk among UC patients can be reduced by regular therapy with 5-ASA medication. Colonoscopic surveillance may be best targeted on those unable to take 5-ASAs (e.g. due to allergy) and those with a positive family history of CRC. [source] Is physician-diagnosed allergic rhinitis a risk factor for the development of asthma?ALLERGY, Issue 8 2010L. Van Den Nieuwenhof To cite this article: van den Nieuwenhof L, Schermer T, Bosch Y, Bousquet J, Heijdra Y, Bor H, van den Bosch W, van Weel C. Is physician-diagnosed allergic rhinitis a risk factor for the development of asthma? Allergy 2010; 65: 1049,1055. Abstract Background:, There is strong evidence that there is a relationship between allergic rhinitis (AR) and asthma, but it is unclear whether there is a causal relation between AR and asthma. The aim of this study was to assess prospectively whether AR is a risk factor for the diagnosis of asthma in a large primary care population. Methods:, We performed a historic cohort study of life-time morbidity that had been recorded prospectively since 1967 in four general practices. Two groups of subjects were selected: (i) patients with diagnosis of AR, (ii) a control group matched using propensity scores. We assessed the risk of physician-diagnosed asthma in patients with physician-diagnosed AR compared to subjects without a diagnosis of AR (controls). Results:, The study population consisted of 6491 subjects (n = 2081 patients with AR). Average study follow-up was 8.4 years. In patients with AR, the frequency of newly diagnosed asthma was 7.6% (n = 158) compared to 1.6% (n = 70) in controls (P < 0.001). After adjusting the effect of AR on asthma diagnosis for registration time, age, gender, eczema and socioeconomic status, having AR was a statistically significant risk factor for asthma (hazard ratio: 4.86, P < 0.001, 95% confidence interval: 3.50,6.73, controls as reference). Conclusion:, A diagnosis of AR was an independent risk factor for asthma in our primary care study population. Having physician-diagnosed AR increased the risk almost fivefold for a future asthma diagnosis. [source] | ||||||||||||||||||||||||||||||||||