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Kinds of Inactive Terms modified by Inactive Selected AbstractsPhotoperiod,Testicular,Immune Interaction in a Seasonal Breeder Indian Palm Squirrel Funambulus pennanti During the Reproductively Inactive and Active PhasesJOURNAL OF NEUROENDOCRINOLOGY, Issue 1 2009R. Ahmad The differential effect of long (LD; 16 : 8 h light/dark), short (SD; 10 : 14 h light/dark) and natural day length (NDL; 12 : 12 h light/dark) during the reproductively inactive (RIP) and active (RAP) phases was assessed in relation to immunity and reproductive function of a tropical rodent Funambulus pennanti. They presented high immunity and low testicular activity during RIP and an opposite during RAP. SD increased spleen and thymus weight, leukocyte and lymphocyte counts, cell mediated immunity [i.e. blastogenic response in terms of percentage stimulation ratio of splenocytes and thymocytes (when challenged with concanavalin A)] and delayed type hypersensitivity to oxazolone. SD during RIP increased the above mentioned parameters and reduced testes weight compared to NDL groups. During RAP, LD reduced all the immunological parameters when compared with NDL and SD experiencing groups of RIP and RAP phases. The LD group reduced the immunological parameters compared to RAP, suggesting that LD had always an inhibitory effect on immune status being independent of reproductive phases. The intensity of the stimulatory effects of SD and inhibitory effects of LD during both reproductive phases was significantly different. We exposed another set of squirrels to the above photoperiodic schedule for prolonged period (30 weeks) during RAP. A clear testicular refractoriness followed by immunorefractoriness was observed in the group experiencing SD and LD for 30 weeks. The photorefractoriness presented by the testes was inversely related to the immunorefractoriness. The peripheral melatonin level of those squirrels reflected the photoperiodic signal perceived by squirrels for immunomodulation and gonadal function, suggesting that immune system and gonadal function might have coevolved. [source] Chemical Approaches to Controlling Intracellular Protein DegradationCHEMBIOCHEM, Issue 1 2005John S. Schneekloth Jr. Inactive. Recent advances have yielded many ways to study proteins by means of inactivation. Traditional methods of genetic knockout are complimented by newer techniques, including RNAi and small molecules that induce proteolysis (see scheme). Although seemingly in competition, these techniques each offer solutions to specific problems in proteomic analysis. [source] Detection of in vitro demineralization of primary teeth using quantitative light-induced fluorescence (QLF)INTERNATIONAL JOURNAL OF PAEDIATRIC DENTISTRY, Issue 3 2002I. A. Pretty Summary. Introduction. With the advent of remineralizing therapies and the new, conservative approach to restoration placement, interest in detecting and monitoring subclinical, precavitated lesions has increased. The increased understanding of clinicians about the processes of primary and secondary prevention and the detection of lesions to which these therapies may be applied, is one of the current goals in caries management. Quantitative light-induced fluorescence (QLF) is a new method for the detection of very early caries. Objectives. To determine the ability of QLF to detect and longitudinally monitor in vitro enamel demineralization. To present the device to the paediatric community and present future in vivo uses of the device. Design. An in vitro study with combined in vivo pilot. Sample and methods. Twelve previously extracted, caries free, primary molars were selected and prepared. Two teeth were randomly selected as controls. Teeth were prepared by gentle pumicing and coating in an acid-resistant nail-varnish, except for an exposed window on the buccal surface. QLF baseline images were taken and the teeth then exposed to a demineralizing solution. Teeth were removed at regular intervals (24, 48, 72, 96, 120, and 144 h), air-dried and QLF images taken. QLF images were analysed by a single, blinded examiner (to control, to length of exposure). Mineral loss, as measured by ,Q, was recorded. Results. Demineralization was noted in all experimental teeth by 48 h, and within 24 h in six teeth. The QLF successfully monitored the increase in mineral loss over time (P < 0·05). The detected lesions were not visible clinically until 144 h and then in only the most severe lesions. No demineralization was detected by QLF in control teeth. The device was user- and patient-friendly in vivo, detecting subclinical lesions. Conclusion. Detection of very early mineral loss and subsequent monitoring of this loss is possible in primary teeth using QLF. The device is well suited to use in paediatric dentistry and offers applications for both clinicians and researchers. The determination of the status of carious lesions (active/inactive) will be possible with readings taken at recall appointments. [source] Quantitative MRI-pathology correlations of brain white matter lesions developing in a non-human primate model of multiple sclerosisNMR IN BIOMEDICINE, Issue 2 2007Erwin L. A. Blezer Abstract Experimental autoimmune encephalomyelitis (EAE) induced with recombinant human myelin/oligodendrocyte glycoprotein in the common marmoset is a useful preclinical model of multiple sclerosis in which white matter lesions can be well visualized with MRI. In this study we characterized lesion progression with quantitative in vivo MRI (4.7,T; T1 relaxation time,±,Gd-DTPA; T2 relaxation time; magnetization transfer ratio, MTR, imaging) and correlated end stage MRI presentation with quantitative ex vivo MRI (formaldehyde fixed brains; T1 and T2 relaxation times; MTR) and histology. The histopathological characterization included axonal density measurements and the numeric quantification of infiltrated macrophages expressing markers for early active [luxol fast blue (LFB) or migration inhibition factor-related protein-14 positive] or late active/inactive [periodic acid Schiff (PAS) positive] demyelinating lesion. MRI experiments were done every two weeks until the monkeys were sacrificed with severe EAE-related motor deficits. Compared with the normal appearing white matter, lesions showed an initial increase in T1 relaxation times, leakage of Gd-DTPA and decrease in MTR values. The progressive enlargement of lesions was associated with stabilized T1 values, while T2 initially increased and stabilized thereafter and MTR remained decreased. Gd-DTPA leakage was highly variable throughout the experiment. MRI characteristics of the cortex and (normal appearing) white matter did not change during the experiment. We observed that in vivo MTR values correlated positively with the number of early active (LFB+) and negatively with late active (PAS+) macrophages. Ex vivo MTR and relaxation times correlated positively with the number of PAS-positive macrophages. None of the investigated MRI parameters correlated with axonal density. Copyright © 2006 John Wiley & Sons, Ltd. [source] Thinking about flagellar oscillationCYTOSKELETON, Issue 8 2009Charles J. Brokaw Abstract Bending of cilia and flagella results from sliding between the microtubular outer doublets, driven by dynein motor enzymes. This review reminds us that many questions remain to be answered before we can understand how dynein-driven sliding causes the oscillatory bending of cilia and flagella. Does oscillation require switching between two distinct, persistent modes of dynein activity? Only one mode, an active forward mode, has been characterized, but an alternative mode, either inactive or reverse, appears to be required. Does switching between modes use information from curvature, sliding direction, or both? Is there a mechanism for reciprocal inhibition? Can a localized capability for oscillatory sliding become self-organized to produce the metachronal phase differences required for bend propagation? Are interactions between adjacent dyneins important for regulation of oscillation and bend propagation? Cell Motil. Cytoskeleton 2008. © 2008 Wiley-Liss, Inc. [source] Requirement for ,B1-crystallin promoter of Xenopus laevis in embryonic lens development and lens regenerationDEVELOPMENT GROWTH & DIFFERENTIATION, Issue 3 2005Nobuhiko Mizuno Regulation of the lens-specific ,B1-crystallin promoter in Xenopus laevis was investigated using transgenic larvae and tadpoles. Comparison of the promoter sequence with that of chicken ,B1-crystallin gene indicates significant sequence similarity over a span of several hundred base pairs starting from the transcriptional start site. Remarkably, PL-1 and PL-2 sequences identified in the chicken promoter as essential binding sites of MAF, Pax6 and Prox1 transcription factors were conserved. Mutations of X (Xenopus) PL-1 and XPL-2 sequences eliminated the promoter activity, indicating a conserved mechanism regulating ,B1-crystallin promoter among vertebrate species. A stepwise deletion of the promoter sequence starting from 2800 bp indicated that the proximal 260 bp directly upstream of the transcription initiation site is sufficient for eliciting lens-specific expression, but the 150 bp promoter sequence is inactive despite it containing the XPL-1 and XPL-2 sequences, suggesting the presence of an additional and essential regulatory sequence located between ,150 and ,260 bp. Activity of the ,B1-crystallin promoter during lens regeneration from cornea was examined using transgenic tadpoles and found to have the same dependence on promoter regions as in embryonic lens development, indicating that gene regulation is largely shared by the two lens-generating processes. [source] Analysis of regulatory elements of E-cadherin with reporter gene constructs in transgenic mouse embryosDEVELOPMENTAL DYNAMICS, Issue 2 2003Marc P. Stemmler Abstract Proper regulation of E-cadherin,mediated cell adhesion is important during early embryonic development and in organogenesis. In mice, E-cadherin is expressed from the fertilized egg onward and becomes down-regulated during gastrulation in mesoderm and its derivatives, but its expression is maintained in all epithelia. E-cadherin promoter analyses led to the identification of binding sites for two transcriptional repressors, Snail and SIP1, which are able to mediate down-regulation in vitro, but little is known about the regulatory elements that govern E-cadherin transcriptional activity in vivo. Here, we compared the developmentally regulated expression of a series of lacZ -reporter transgenes fused to different sequences of the murine E-cadherin gene between ,6 kb, including the promoter, and +16 kb, covering one third of intron 2. Four different segments with distinct regulatory properties were identified. The promoter fragment from +0.1 to ,1.5 kb remains inactive in most cases but occasionally induces ectopic expression in mesodermal tissues, although it contains binding sites for the repressors Snail and SIP1. This promoter fragment also lacks positive elements needed for the activation of transcription in ectoderm and endoderm. Sequences from ,1.5 to ,6 kb harbor regulatory elements for brain-specific expression and, in addition, insulator or silencer elements, because they are consistently inactive in the mesoderm. Only if sequences from +0.1 to +11 kb are combined with the promoter fragments is E-cadherin,specific transgene expression observed in endoderm and certain epithelia. Sequences between +11 and +16 kb contain cis -active elements that generally enhance transcription. Our analyses show that E-cadherin expression is governed by a complex interplay of multiple regulatory regions dispersed throughout large parts of the locus. Developmental Dynamics 227:238,245, 2003. © 2003 Wiley-Liss, Inc. [source] Pharmacokinetics of dipeptidylpeptidase-4 inhibitorsDIABETES OBESITY & METABOLISM, Issue 8 2010A. J. Scheen Type 2 diabetes (T2DM) is a complex disease combining defects in insulin secretion and insulin action. New compounds have been developed for improving glucose-induced insulin secretion and glucose control, without inducing hypoglycaemia or weight gain. Dipeptidylpeptidase-4 (DPP-4) inhibitors are new oral glucose-lowering agents, so-called incretin enhancers, which may be used as monotherapy or in combination with other antidiabetic compounds. Sitagliptin, vildaglipin and saxagliptin are already on the market in many countries, either as single agents or in fixed-dose combined formulations with metformin. Other DPP-4 inhibitors, such as alogliptin and linagliptin, are currently in late phase of development. The present paper summarizes and compares the main pharmacokinetics (PK) properties, that is, absorption, distribution, metabolism and elimination, of these five DPP-4 inhibitors. Available data were obtained in clinical trials performed in healthy young male subjects, patients with T2DM, and patients with either renal insufficiency or hepatic impairment. PK characteristics were generally similar in young healthy subjects and in middle-aged overweight patients with diabetes. All together gliptins have a good oral bioavailability which is not significantly influenced by food intake. PK/pharmacodynamics characteristics, that is, sufficiently prolonged half-life and sustained DPP-4 enzyme inactivation, generally allow one single oral administration per day for the management of T2DM; the only exception is vildagliptin for which a twice-daily administration is recommended because of a shorter half-life. DPP-4 inhibitors are in general not substrates for cytochrome P450 (except saxagliptin that is metabolized via CYP 3A4/A5) and do not act as inducers or inhibitors of this system. Several metabolites have been documented but most of them are inactive; however, the main metabolite of saxagliptin also exerts a significant DPP-4 inhibition and is half as potent as the parent compound. Renal excretion is the most important elimination pathway, except for linagliptin whose metabolism in the liver appears to be predominant. PK properties of gliptins, combined with their good safety profile, explain why no dose adjustment is necessary in elderly patients or in patients with mild to moderate hepatic impairment. As far as patients with renal impairment are concerned, significant increases in drug exposure for sitagliptin and saxagliptin have been reported so that appropriate reductions in daily dosages are recommended according to estimated glomerular filtration rate. The PK characteristics of DPP-4 inhibitors suggest that these compounds are not exposed to a high risk of drug,drug interactions. However, the daily dose of saxagliptin should be reduced when coadministered with potent CYP 3A4 inhibitors. In conclusion, besides their pharmacodynamic properties leading to effective glucose-lowering effect without inducing hypoglycaemia or weight gain, DPP-4 inhibitors show favourable PK properties, which contribute to a good efficacy/safety ratio for the management of T2DM in clinical practice. [source] The association of physical activity and depression in Type 2 diabetesDIABETIC MEDICINE, Issue 10 2008Z. Lysy Abstract Aims Physical inactivity and depressed mood are both associated with a higher likelihood of diabetes-related complications; the association between physical activity and depressed mood in Type 2 diabetes has not been reviewed previously. We have reviewed (i) the strength of this association and (ii) the impact of depression-specific management and physical activity interventions on mood and activity levels in overweight adults with Type 2 diabetes. Methods Studies published between January 1996 and September 2007 were identified (Ovid - medline, Psych- Info and embase) using pertinent search terms (keyword/title). Results Of the 12 studies included (10 cross-sectional, two trials), most employed a standardized questionnaire for depressed mood but only one item for physical activity. In adults with Type 2 diabetes, the inactive are 1.72 to 1.75 times more likely to be depressed than the more active; the depressed are 1.22 to 1.9 times more likely to be physically inactive than the non-depressed. Two randomized trials demonstrated that a depression management programme improved mood, but only one demonstrated increased physical activity. Conclusions Studies to date suggest an association between depressed mood and physical inactivity in adults with Type 2 diabetes, although objective measures of physical activity have not been employed. Depression-specific management may improve mood and possibly activity. A trial comparing the impact of depression-specific management compared with exercise intervention on depressed mood and activity in Type 2 diabetes is justified. [source] Prevalence and projections of diabetes and pre-diabetes in adults in Sri Lanka,Sri Lanka Diabetes, Cardiovascular Study (SLDCS)DIABETIC MEDICINE, Issue 9 2008P. Katulanda Abstract Aims To determine the prevalence of diabetes mellitus and pre-diabetes (impaired fasting glucose and impaired glucose tolerance) in adults in Sri Lanka. Projections for the year 2030 and factors associated with diabetes and pre-diabetes are also presented. Methods This cross-sectional study was conducted between 2005 and 2006. A nationally representative sample of 5000 adults aged , 18 years was selected by a multi-stage random cluster sampling technique. Fasting plasma glucose was tested in all participants and a 75-g oral glucose tolerance test was performed in non-diabetic subjects. Prevalence was estimated for those > 20 years of age. Results Response rate was 91% (n = 4532), males 40%, age 46.1 ± 15.1 years (mean ± standard deviation). The age,sex standardized prevalence (95% confidence interval) of diabetes for Sri Lankans aged , 20 years was 10.3% (9.4,11.2%) [males 9.8% (8.4,11.2%), females 10.9% (9.7,12.1%), P = 0.129). Thirty-six per cent (31.9,40.1%) of all diabetic subjects were previously undiagnosed. Diabetes prevalence was higher in the urban population compared with rural [16.4% (13.8,19.0%) vs. 8.7% (7.8,9.6%); P < 0.001]. The prevalence of overall, urban and rural pre-diabetes was 11.5% (10.5,12.5%), 13.6% (11.2,16.0%) and 11.0% (10.0,12.0%), respectively. Overall, 21.8% (20.5,23.1%) had some form of dysglycaemia. The projected diabetes prevalence for the year 2030 is 13.9%. Those with diabetes and pre-diabetes compared with normal glucose tolerance were older, physically inactive, frequently lived in urban areas and had a family history of diabetes. They had higher body mass index, waist circumference, waist,hip ratio, systolic/diastolic blood pressure, low-density lipoprotein cholesterol and triglycerides. Insulin was prescribed to 4.4% (2.7,6.1%) of all diabetic subjects. Conclusions One in five adults in Sri Lanka has either diabetes or pre-diabetes and one-third of those with diabetes are undiagnosed. [source] Increased levels of monokine induced by interferon-, (Mig) in the vitreous of patients with diabetic retinopathyDIABETIC MEDICINE, Issue 7 2008Y. Wakabayashi Abstract Aim To determine the intravitreous concentration of monokine induced by interferon-, (Mig) in patients with diabetic retinopathy (DR) and the relation between Mig and vascular endothelial growth factor (VEGF). Research design and methods Vitreous samples were obtained at the time of vitrectomy from 41 eyes of 38 DR patients (30 with active DR and 11 with inactive DR) and from 15 eyes of 15 non-diabetic patients who had macular disease (control subjects). Human Mig and VEGF were quantified using a FACS Caliber® flow cytometer. Results The vitreous concentration of Mig was increased significantly in patients with both active and inactive DR [148.0 (31.6,997.2; median range) and 82.3 (25.7,347.7) pg/ml, respectively] compared with control subjects [21.0 (5.2,74.3) pg/ml; P < 0.0001 and P < 0.001, respectively]. In DR patients, a significant (P < 0.01) correlation was observed between vitreous concentrations of Mig and VEGF. Conclusion Our results suggest that Mig may play an important role in the pathogenesis of DR and works in consort with VEGF in the progression of pathological angiogenesis in DR. [source] Influence of lipophilicity and stereochemistry at the C7 position on the cardioprotective and antioxidant effect of ginkgolides during rat heart ischemia and reperfusion,DRUG DEVELOPMENT RESEARCH, Issue 3 2005Ludovic Billottet Abstract The extent to which the cardioprotective effect of ginkgolides is related to their lipophilicity rather than to their anti-platelet activating factor (PAF) effect was addressed in isolated rat hearts submitted to ischemia and reperfusion. A new derivative of ginkgolide C (1), the 7-,- O -(4-methylphenyl) ginkgolide C (4) was synthesized and compared to 7- O -(4-methylphenyl) ginkgolide C (2) that had the same absolute configuration at C7 as 1 for its lipophilicity, anti-PAF activity, and cardioprotective and antioxidant effects. Using reversed-phase high-performance liquid chromatography HPLC, 4 and 2 were found to be significantly more lipophilic (i.e., log kw of 3.42±0.05 and 3.64±0.07, respectively) than 1 (1.15±0.03) and the strong PAF inhibitor ginkgolide B (GkB; 1.65±0.03). The anti-PAF activities (IC50 values in ,M) were 8.2, 17.1, and 2.2 for 4, 1, and GkB, respectively, while 2 was inactive. In preischemic and/or reperfused hearts perfused with ginkgolides at 0.7 ,M: (i) 2 and 4 were more efficient in improving postischemic hemodynamic and metabolic recovery than 1, (ii) a key-step in cardioprotection occurred during ischemia where 2 and 4 limited myocardial ATP depletion and contracture development, (iii) a strong anti-lipoperoxidant effect was observed with 2 and 4, but not 1. In vivo administration of 2 to rats (4 mg/kg/day for 20 days) was more effective than that of 1 regarding ischemic heart protection, suggesting a positive role for lipophilicity. It was concluded that a high lipophilicity is not an absolute prerequisite for a strong anti-PAF effect for ginkgolides, whereas it appears essential for cardioprotection. Drug Dev. Res. 64:157,171, 2005. © 2005 Wiley-Liss, Inc. [source] Characterization and expression of ATP P2X4 receptor from embryonic chick skeletal muscleDRUG DEVELOPMENT RESEARCH, Issue 1 2001Xuenong Bo Abstract Previous pharmacological experiments have indicated the existence of ATP P2X receptors in chick embryonic skeletal muscles. In this study we cloned a P2X4 -like cDNA encoding a protein of 385 amino acids, which shares 75% and 76% identity with rat and human P2X4 receptors, respectively. Functional studies of this cP2X4 receptor expressed in Xenopus oocytes showed that ATP induced a fast inward current, which was partially desensitized upon prolonged application of ATP. The ATP-induced currents were concentration-dependent, with an EC50 of 9.5 ,M. Adenosine 5,- O -(thio)triphosphate and 2-methylthioATP very weak agonists. ,,,-methyleneATP was almost inactive. In contrast to their potentiating effects on recombinant rat P2X4 receptors, both suramin and pyridoxalphosphate-6-azophenyl-2,,4,-disulfonic acid partially blocked ATP-induced currents. TrinitrophenylATP was able to block ATP-induced response completely, with an IC50 of 4.7 ,M. Northern blot and RT-PCR analysis showed that cP2X4 mRNAs were mainly expressed in skeletal muscle, brain, and gizzard of day 10 chick embryos. Lower levels of expression were also detected in liver, heart, and retina. Whole-mount in situ hybridization showed that cP2X4 mRNAs were expressed in the brain, spinal cord, notochord, gizzard, and skeletal muscle. The physiological functions of cP2X4 receptors in embryonic skeletal muscle remain unclear at present. Drug Dev. Res. 53:22,28, 2001. © 2001 Wiley-Liss, Inc. [source] The termination of the last major phase of aeolian sand movement, coastal dunefields, DenmarkEARTH SURFACE PROCESSES AND LANDFORMS, Issue 7 2006Lars B. Clemmensen Abstract Optically stimulated luminescence (OSL) dating of sand samples from stabilized (or inactive) coastal dunes in Denmark provides information on the age of the termination phase of the last major aeolian activity period. A total of 26 sand samples were taken from four different coastal dunefields around the North Sea, Skagerrak and Kattegat coasts of Denmark. The OSL dates indicate that the last major phase of aeolian activity terminated between ad 1860 and 1905. Most of the dunes examined in this study were active around 1820, during a period documented to have been very stormy. A dune management scheme started around 1792, and this no doubt was a major cause of dunefield stabilization, but an overall decline of storminess, particularly spring and summer storminess, around the end of the 19th century must also have contributed to the increasing inactivity of coastal dunes. The new OSL dates on aeolian sand movement agree well with historical data and data from topographic maps on dune movement. This agreement supports the observation from earlier work that OSL dating of recent aeolian sand movement is accurate over the last few decades to centuries. Copyright © 2005 John Wiley & Sons, Ltd. [source] Companion planting , behaviour of the cabbage root fly on host plants and non-host plantsENTOMOLOGIA EXPERIMENTALIS ET APPLICATA, Issue 1 2005Kate Morley Abstract Six-hundred individual female cabbage root flies (Delia radicum L.) (Diptera: Anthomyiidae) were each observed for 20 min under laboratory conditions to record how they behaved after landing on a host or a non-host plant. Fly movements were recorded on host plants [cabbage ,Brassica oleracea var. capitata (Cruciferae)] and non-host plants [clover ,Trifolium subterraneum L. (Papilionaceae)] surrounded by bare soil and on cabbage surrounded by clover. The most frequently observed behaviours made by the flies were (1) hops/spiral flights and (2) walks/runs. In the bare soil situation, the 50 individual flies observed in each treatment made 66 hops/spiral flights on the cabbage and 94 on the clover. When the two plants were tested together the movements were not additive as, instead of the expected 160 hops/spiral flights in the mixed plant treatment, the flies made 210 hops/spiral flights when they landed initially on cabbage but only 130 when they landed initially on clover. Few of the flies that landed initially on clover moved onto the host plant, even though the host plant was only a few centimetres away. The duration of the individual walks and runs made by the cabbage root flies were similar on both the host and non-host plants. The only differences were the numbers of walks/runs made and the time the flies remained inactive. On the host plants, the females made four walks/runs, each of about 12 s duration, interspersed by rest periods that totalled 1.5 min. In contrast, on the non-host plants the females made 10 walks/runs, each of about 9 s duration, interspersed by rest periods that totalled 7 min. Therefore, after landing on a plant, the flies, on average, left the host plant after 2.25 min and the non-host plant after 8.5 min. Our conclusion is that the protracted time spent on the non-host plants is the mechanism that disrupts insects from finding host plants in diverse plantings. Hence, the flies were arrested by non-host plants rather than being repelled or deterred as suggested in earlier studies. [source] In situ measurement of methane fluxes and analysis of transcribed particulate methane monooxygenase in desert soilsENVIRONMENTAL MICROBIOLOGY, Issue 10 2009Roey Angel Summary Aerated soils are a biological sink for atmospheric methane. However, the activity of desert soils and the presence of methanotrophs in these soils have hardly been studied. We studied on-site atmospheric methane consumption rates as well as the diversity and expression of the pmoA gene, coding for a subunit of the particulate methane monooxygenase, in arid and hyperarid soils in the Negev Desert, Israel. Methane uptake was only detected in undisturbed soils in the arid region (,90 mm year,1) and vertical methane profiles in soil showed the active layer to be at 0,20 cm depth. No methane uptake was detected in the hyperarid soils (,20 mm year,1) as well as in disturbed soils in the arid region (i.e. agricultural field and a mini-catchment). Molecular analysis of the methanotrophic community using terminal restriction fragment length polymorphism (T-RFLP) and cloning/sequencing of the pmoA gene detected methanotrophs in the active soils, whereas the inactive ones were dominated by sequences of the homologous gene amoA, coding for a subunit of the ammonia monooxygenase. Even in the active soils, methanotrophs (as well as in situ activity) could not be detected in the soil crust, which is the biologically most important layer in desert soils. All pmoA sequences belonged to yet uncultured strains. Transcript analysis showed dominance of sequences clustering within the JR3, formerly identified in Californian grassland soils. Our results show that although active methanotrophs are prevalent in arid soils they seem to be absent or inactive in hyperarid and disturbed arid soils. Furthermore, we postulate that methanotrophs of the yet uncultured JR3 cluster are the dominant atmospheric methane oxidizers in this ecosystem. [source] New ways to break an old bond: the bacterial carbon,phosphorus hydrolases and their role in biogeochemical phosphorus cyclingENVIRONMENTAL MICROBIOLOGY, Issue 10 2007John P. Quinn Summary Phosphonates are organophosphorus molecules that contain the highly stable C,P bond, rather than the more common, and more labile, C,O,P phosphate ester bond. They have ancient origins but their biosynthesis is widespread among more primitive organisms and their importance in the contemporary biosphere is increasingly recognized; for example phosphonate-P is believed to play a particularly significant role in the productivity of the oceans. The microbial degradation of phosphonates was originally thought to occur only under conditions of phosphate limitation, mediated exclusively by the poorly characterized C,P lyase multienzyme system, under Pho regulon control. However, more recent studies have demonstrated the Pho-independent mineralization by environmental bacteria of three of the most widely distributed biogenic phosphonates: 2-aminoethylphosphonic acid (ciliatine), phosphonoacetic acid, and 2-amino-3-phosphonopropionic acid (phosphonoalanine). The three phosphonohydrolases responsible have unique specificities and are members of separate enzyme superfamilies; their expression is regulated by distinct members of the LysR family of bacterial transcriptional regulators, for each of which the phosphonate substrate of the respective degradative operon serves as coinducer. Previously no organophosphorus compound was known to induce the enzymes required for its own degradation. Whole-genome and metagenome sequence analysis indicates that the genes encoding these newly described C,P hydrolases are distributed widely among prokaryotes. As they are able to function under conditions in which C,P lyases are inactive, the three enzymes may play a hitherto-unrecognized role in phosphonate breakdown in the environment and hence make a significant contribution to global biogeochemical P-cycling. [source] Structure,activity relationships for gene activation oestrogenicity: Evaluation of a diverse set of aromatic chemicalsENVIRONMENTAL TOXICOLOGY, Issue 1 2002T. Wayne Schultz Abstract Structure,activity relationships for oestrogenicity were developed based on 120 aromatic chemicals evaluated in the Saccharomyces cerevisiae -based Lac -Z reporter assay. Relative gene activation was compared to 17,-estradiol and varied over eight orders of magnitude. Analysis of the data compared to 17,-estradiol identified three structural criteria that were related to xenoestrogen activity and potency: (1) the hydrogen-bonding ability of the phenolic ring mimicking the A-ring, (2) a hydrophobic centre similar in size and shape to the B- and C-rings, and (3) a hydrogen-bond donor mimicking the 17,-hydroxyl moiety of the D-ring, especially with an oxygen-to-oxygen distance similar to that between the 3- and 17,-hydroxyl groups of 17,-estradiol. Binding data were segregated into activity clusters including strong, moderate, weak, and detectable gene expression, and those compounds that were inactive. The hydrogen-bonding ability of hydroxy group in the 3-position on 17,-estradiol was observed to be essential for gene activation. Compounds with a 4-hydroxyl substituted benzene ring and a hydrophobic moiety of size and shape equivalent to the B-ring of 17,-estradiol were generally observed to be weakly active compounds. Moderately active compounds have a 4-hydroxyl substituted benzene ring with a hydrophobic moiety equivalent in size and shape to the B- and C-ring of 17,-estradiol, or have a high hydrogen-bond donor capacity owing to the presence of halogens on a nonphenolic ring. Strongly active compounds, similar to 4,4,-diethylethylene bisphenol (DES), possess the same hydrophobic ring structure as described for moderately active compounds and an additional hydroxyl group with an oxygen-to-oxygen distance close to that exhibited by the 3- and 17-hydroxyl groups of 17,-estradiol. © 2002 by Wiley Periodicals, Inc. Environ Toxicol 17: 14,23, 2002 [source] Physical activity in adolescence and smoking in young adulthood: a prospective twin cohort studyADDICTION, Issue 7 2007Urho M. Kujala ABSTRACT Aims To control for familial confounds, we studied the association between adolescent physical activity and later smoking in twin siblings discordant for their baseline physical activity. Design and measurements In this prospective population-based twin study, we asked whether persistent physical activity/inactivity in adolescence (assessed at 16, 17 and 18.5 years) predicted questionnaire-reported daily smoking at ages 22,27. Twins who, on the three baseline questionnaires, consistently reported frequent leisure physical activity (more than three times weekly) were classified as persistent exercisers, those who exercised less than three times monthly were called persistently inactive, others were occasional exercisers. Setting Finland. Participants A total of 4240 individuals, including 1870 twin pairs. Findings In analyses of individual twins, compared to persistent activity, persistent physical inactivity predicted increased risk of daily smoking (age- and sex-adjusted odds ratio 5.53, 95% confidence interval 3.88,7.88, P < 0.001). The risk remained elevated even after excluding all those who had smoked 50 cigarettes or more life-time at baseline and adjusted for educational level in adolescence. In within-pair analyses compared to the active members of discordant twin pairs, the physically inactive co-twins had increased risk of future daily smoking (sex-adjusted odds ratio 3.39, 95% confidence interval 1.56,7.39, P = 0.002). Conclusions Persistent physical inactivity in adolescence relates to adult smoking, even after familial factors are taken into account. [source] Evaluating polycyclic aromatic hydrocarbons using a yeast bioassayENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2007Abeer Alnafisi Abstract Sixteen polycyclic aromatic hydrocarbons (PAHs) were evaluated for the ability to activate aryl hydrocarbon (Ah) receptor signaling in a yeast-based bioassay. Individual PAHs were classified as inactive or as weakly, moderately, or strongly active based on induction of human Ah receptor signaling. Indeno[1,2,3- cd]pyrene, chrysene, benzo[a]anthracene, benzo[a]pyrene, benzo[j]fluoranthene, and benzo[k]fluoranthene were the most potent activators of human Ah receptor signaling. Various mixtures of PAHs had additive or synergistic effects in the bioassay. Environmental samples from the New Orleans (Louisiana, USA) and Detroit (Michigan, USA) areas that were previously analyzed for PAH composition and quantity were tested in this bioassay. Weak but statistically significant relationships were found when the analytically measured levels of PAHs were correlated with sample dilutions that gave 25% effective concentration signaling levels in the Ah receptor assay. We conclude that this Ah receptor signaling assay may be useful for preliminary biomonitoring of samples for PAHs and other Ah receptor ligands. [source] Biological measurement of estrogenic activity in urine and bile conjugates with the in vitro ER-CALUX reporter gene assayENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2002Juliette Legler Abstract Although estrogens are excreted as biologically inactive conjugates, they can be reconverted to an active form, possibly by bacteria. A simple method was developed to deconjugate estrogen metabolites present in human urine and fish bile back to active estrogens by enzymatic hydrolysis with ,-glucuronidase or live Escherichia coli cells. Deconjugated extracts were tested for estrogenic activity in the in vitro stable estrogen receptor,mediated chemical-activated luciferase gene expression (ER-CALUX) assay. Estrogen glucuronides in urine obtained from human males and females were effectively converted to active forms after incubation with ,-glucuronidase or E. coli. The highest estrogenic activity was found in deconjugated metabolites from urine of a pregnant woman, in which levels up to 3,000 nmol estradiol equivalents per liter of urine were found after overnight incubation of urine with E. coli. Bile sampled from male bream and flounder from various freshwater and marine locations was also deconjugated and a good correlation was found between high biliary estrogenic activity and elevated levels of xenoestrogenic activity in surface water as well as in plasma vitellogenin. Therefore, the measurement of deconjugated bile could form a useful (indirect) biomarker for internal dose of xenoestrogens in male fish. [source] Effects of Circadian Regulation and Rest,Activity State on Spontaneous Seizures in a Rat Model of Limbic EpilepsyEPILEPSIA, Issue 5 2000Mark Quigg Summary: Purpose: Circadian regulation via the suprachiasmatic nuclei and rest,activity state may influence expression of limbic seizures. Methods: Male rats (n = 14) were made epileptic by electrical stimulation of the hippocampus, causing limbic status epilepticus and subsequent seizures. We monitored seizures with intrahippocampal electrodes in 12,12-h light/dark (LD) cycles and in continuous dark (DD). We used radiotelemetry monitoring of activity to measure state and body temperature to determine circadian phase. Cosinor analysis and ,2 tests determined whether seizures occurred rhythmically when plotted by phase. State was defined as inactive or active in 10-min epochs based on whether activity count was below or above a cut-off value validated from video observation. Results: In LD, the peak seizure occurrence was 14:59 h after circadian temperature peak (95% confidence limit, 13:37,16:19). Phasic seizure occurrence persisted in DD for 14:05 (12:31,15:38), p < 0.0001, against uniform mean distribution. In LD, 14,787 epochs contained 1,268 seizures; seizures preferentially occurred during inactive epochs (965 observed, 878 expected in proportion to the overall distribution of inactive versus active epochs; p < 0.001). In DD, 20,664 epochs contained 1,609 seizures; seizures had no preferential occurrence by state (999 observed, 1,025 expected; p = 0.16). Conclusions: Limbic seizures occurred with an endogenous circadian rhythm. Seizures preferentially struck during inactivity during entrainment to the light,dark cycle. [source] Temporal Variation in Cleanerfish and Client Behaviour: Does It Reflect Ectoparasite Availability?ETHOLOGY, Issue 6 2003Isabelle M. Côté We tested the importance of ectoparasites as the proximate cause of cleaning interactions by comparing the activity of Caribbean cleaning gobies (Elacatinus evelynae) and of their clients during three daily periods (early morning, midday, and late afternoon) in which ectoparasite availability varied naturally. Emergence from the benthos of gnathiid isopod larvae, the main target of cleaning goby predation, was higher at night, when cleaners were inactive, than during the day. As a result, overall ectoparasite loads on client fish tended to be higher in the morning. Inspection bouts by cleaning gobies were longest in the morning, but also at midday when ectoparasite availability on clients was lower. Client fish were observed at cleaning stations most often in the afternoon, when they harboured few ectoparasites, but they were more likely to adopt incitation poses, which increase the likelihood of being cleaned, in the morning than later in the day. Most cleaner and client behaviours therefore did not change predictably in response to natural diurnal variation in ectoparasite availability. Our study suggests that the ultimate and proximate causes of cleaning behaviour need not necessarily coincide. [source] IRAK-4 kinase activity-dependent and -independent regulation of lipopolysaccharide-inducible genesEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2008Magdalena Koziczak-Holbro Abstract IRAK-4 kinase inactive (IRAK-4 KD) knock-in mice display defects in TLR- and IL-1 receptor signaling and are resistant to LPS-induced shock. In the present study we examined the LPS-induced response in IRAK-4 KD mice in more detail. We show that IRAK-4 kinase activity is required for certain aspects of TLR-mediated signaling but not for others. We found that IRAK-4 KD cells displayed reduced JNK and p38 signaling, while NF-,B was activated to a normal level but with delayed kinetics compared to wild-type cells. TLR4-mediated IRF3 activation was intact in these cells. Comprehensive analysis of expression of LPS-inducible genes by microarray demonstrated that IRAK-4 KD cells were severely impaired in the expression of many pro-inflammatory genes, suggesting their dependence on IRAK-4 kinase activity. In contrast, the expression of a subset of LPS-induced genes of anti-viral response was not affected by IRAK-4 kinase deficiency. Additionally, we demonstrate that LPS-activated early expression and production of some cytokines, e.g., TNF-,, is partially induced in the absence of IRAK-4 kinase activity. This suggests that the partially unaffected TLR4-mediated signaling could still drive expression of these genes in early phases and that IRAK-4 kinase activity is important for a more sustained anti-bacterial response. See accompanying commentary http://dx.doi.org/10.1002/eji.200838161 [source] Apoptosis via the B cell antigen receptor requires Bax translocation and involves mitochondrial depolarization, cytochrome C release, and caspase-9 activationEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 7 2004Eric Eldering Abstract Various routes to apoptosis can be active during B cell development. In a model system of mature B cells, differences in caspase-3 processing have suggested that antigen receptor (BCR)-mediated apoptosis may involve a zVAD-insensitive initiator protease(s). In search of the events leading to caspase-3 activation, we now establish that both CD95- and BCR-mediated apoptosis depend on Bax activation and cytochrome C (cytC) release. Nevertheless, the timing and caspase-dependence of mitochondrial membrane depolarization differed considerably after CD95- or BCR-triggering. To delineate events subsequent to cytC release, we compared apoptosis induced via BCR triggering and via direct mitochondrial depolarization by CCCP. In both cases, partial processing of caspase-3 was observed in the presence of zVAD. By expression in 293 cells we addressed the potential of candidate initiator caspases to function in the presence of zVAD, and found that caspase-9 efficiently processed caspase-3, while caspase-2 or ,8 were inactive. Finally, retroviral expression of dominant-negative caspase-9 inhibited both CD95- and BCR-mediated apoptosis. In conclusion, we obtained no evidence for involvement of a BCR-specific protease. Instead, our data show for the first time that the BCR-signal causes Bax translocation, followed by mitochondrial depolarization, and cytC release. Subsequent caspase-9 activation can solely account for events further downstream. [source] Proximal changes in signal transduction that modify CD8+ T cell responsiveness in vivoEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 9 2003Séverine Guillaume Abstract The antigen dose conditions the functional properties of CD8+ T cells generated after priming. At relatively low antigen doses, efficient memory T cells may be generated, while high antigen doses lead to tolerance. To determine the mechanisms leading to such different functional outcomes, we compared the proximal TCR signal transduction of naive cells, to that of memory or high-dose tolerant cells generated in vivo. In vivo activation led to the constitutive phosphorylation of CD3,, recruiting Zap70, in both memory and tolerant cells. In tolerant cells, these phenomena were much more marked, the CD3, and , chains no longer associated, and the Src kinases p56Lck and p59Fyn were inactive. Therefore, when the antigen load overcomes the capacities of immune control, a new mechanism intervenes to block signal transduction: the recruitment of Zap70 to CD3, becomes excessive, leading to TCR complex destabilization, Src kinase dysfunction, and signal arrest. [source] The ester-bonded palmitoyl side chains of Pam3CysSerLys4 lipopeptide account for its powerful adjuvanticity to HLA class,I-restricted CD8+ T,lymphocytesEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 7 2003Anca Reschner Abstract Molecularly defined adjuvants are urgently required to implement immunization protocols by which CD8+ T,cells induction is envisaged. We show here that the synthetic lipopeptide Pam3CysSerLys4 (P3CSK4) strongly enhances the expansion of antigen-specific IFN-,+CD8+ cells in vitro. These effects critically depend on the presence of two ester-bonded palmitoylated side chains. In fact, T,cell expansion is impaired in the presence of derivatives bearing two non-palmitoylated fatty acid chains, while derivatives with only one amide-bonded palmitoylated residue are completely inactive and behave like the non-lipidated peptide backbone. P3CSK4 is not mitogenic for T,lymphocytes and can modulte DC immune biological properties. Indeed, doses as low as 100,ng/ml increase CD86, CD83 and CD40 surface expression on DC, fail to induce CCR7, and trigger a defined pattern of soluble factors associated to immune effector functions. In particular, substantial amounts of TNF-,, IL-6, CCL2 and CXCL10, in the absence of IFN-,, IFN-,, IL-15, IL-12p70 and CX3CL1, can be measured. Accordingly, antigen-specific CD8+ T,cells expanded in vitro express CCR2 and CXCR3 chemokine receptors. Altogether our data suggest that human DC are able to respond to chemically different synthetic lipopeptide analogs and that optimal adjuvanticity to CD8+ T,cell induction is achieved by the palmitoylated structures. [source] Non-Tethered Organometallic Phosphonate Inhibitors for Lipase Inhibition: Positioning of the Metal Center in the Active Site of Cutinase,,EUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 28 2008Cornelis A. Kruithof Abstract Organometallic NCN-pincer complexes, bearing either a p -nitrophenyl phosphonate ester or a phosphonic acid group directly attached to the aromatic ring of the pincer complex, were synthesized. These compounds were tested as covalent inhibitors for the lipase cutinase. In a stoichiometric reaction of the NCN-pincer platinum phosphonate p -nitrophenyl ester 2 with cutinase, a 94,% conversion to the protein,pincer metal complex hybrid was obtained in 48 h. The NCN-pincer metal phosphonic acid derivatives (3, 4) appeared to be inactive as cutinase inhibitors. In contrast to our previous work which entails propyl tethered phosphonate esters connected to pincer metal complexes, the presented strategy allows positioning of metal complexes inside the active site of lipases. This opens up the possibility for fine-tuning the chemical environment (second coordination sphere) around a synthetic metal center inside the pocket of an enzyme for diagnostic and catalytic purposes.(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008) [source] Structure, Electrochemistry and Hydroformylation Catalytic Activity of the Bis(pyrazolylborato)rhodium(I) Complexes [RhBp(CO)P] [P = P(NC4H4)3, PPh3, PCy3, P(C6H4OMe-4)3]EUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 7 2004Anna M. Trzeciak Abstract Rhodium complexes of formula [RhBp(CO)P] [Bp = bis(pyrazolylborate), P = P(NC4H4)31, PPh32, PCy33, P(C6H4OMe-4)34] have been prepared by exchange of the acetylacetonate (acac,) ligand in [Rh(acac)(CO)P] complexes. The spectroscopic and electrochemical properties as well as X-ray data of [Rh(acac)(CO)P] and [RhBp(CO)P] complexes have been compared with the aim to estimate the relative donor properties of both anionic ligands (acac, and Bp,). The cyclic voltammetric results indicate that the Bp, ligand behaves as a much stronger electron donor than acac, and a value of the Lever EL ligand parameter identical to that of the pyrazolate ligand (,0.24 V vs. NHE for each coordinating arm) is proposed for the bis- and tris(pyrazolyl)borate ligands, whereas P(C6H4OMe-4)3 is also shown to have an identical EL value (0.69 V) to that of P(NC4H4)3. An improved linear relationship between the oxidation potential and the sum of the ligand EL values for square-planar RhI complexes is also obtained and adjusted values for the Lever SM and IM parameters for the RhI/RhII redox couple are given. The trans influence of phosphanes was not observed in crystals of complexes 2 and 3, in contrast to analogous acetylacetonato complexes in which the Rh,O bonds differ by ca. 0.04,0.06 Å. Complexes 1,4 are very attractive precursors for hydroformylation catalysts and yields of aldehydes of 80,87% have been obtained with all complexes without extra phosphane as co-catalyst. During the hydroformylation reaction, however, small amounts of a catalytically inactive [RhBp(CO)2] complex were formed. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source] Tissue-type plasminogen activator-plasmin-BDNF modulate glutamate-induced phase-shifts of the mouse suprachiasmatic circadian clock in vitroEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 8 2009Xiang Mou Abstract The mammalian circadian clock in the suprachiasmatic nucleus (SCN) maintains environmental synchrony through light signals transmitted by glutamate released from retinal ganglion terminals. Brain-derived neurotrophic factor (BDNF) is required for light/glutamate to reset the clock. In the hippocampus, BDNF is activated by the extracellular protease, plasmin, which is produced from plasminogen by tissue-type plasminogen activator (tPA). We provide data showing expression of proteins from the plasminogen activation cascade in the SCN and their involvement in circadian clock phase-resetting. Early night glutamate application to SCN-containing brain slices resets the circadian clock. Plasminogen activator inhibitor-1 (PAI-1) blocked these shifts in slices from wild-type mice but not mice lacking its stabilizing protein, vitronectin (VN). Plasmin, but not plasminogen, prevented inhibition by PAI-1. Both plasmin and active BDNF reversed ,2 -antiplasmin inhibition of glutamate-induced shifts. ,2 -Antiplasmin decreased the conversion of inactive to active BDNF in the SCN. Finally, both tPA and BDNF allowed daytime glutamate-induced phase-resetting. Together, these data are the first to demonstrate expression of these proteases in the SCN, their involvement in modulating photic phase-shifts, and their activation of BDNF in the SCN, a potential ,gating' mechanism for photic phase-resetting. These data also demonstrate a functional interaction between PAI-1 and VN in adult brain. Given the usual association of these proteins with the extracellular matrix, these data suggest new lines of investigation into the locations and processes modulating mammalian circadian clock phase-resetting. [source] | |||||||||||||||||||||||||||||||||||||