Sample records for halobacterium halobium r1

  1. Photoenergy transduction in halobacterium halobium

    Stoeckenius, W.; Bogomolni, R. A.


    The structure and function of a rhodopsin-like pigment bacteriorhodopsin, discovered and isolated from the membranes of the halophile Halobacterium halobium, were studied. Intermediatesm appearing in the cyclic photoreaction that drives the proton translocation, were spectroscopically characterized. The charge translocation in membrane monolayers and mulitlayers placed between electrodes were kinetically resolved. A model was developed for the proton translocation process, in which the isomerization of the retinal Schiff base decreases its pK to drive the proton off and simultaneaouly changes the connectivity from the cytoplasmic surface to the external surface. The stoichiometry of proton pumping in intact cells and the effect of the light generated electrochemical potential on the kinetics of the photoreaction cycle and the synthesis of ATP were investigated.

  2. Light-driven solute transport in Halobacterium halobium

    Lanyi, J. K.


    The cell membrane of Halobacterium halobium exhibits differential regions which contain crystalline arrays of a single kind of protein, termed bacteriorhodopsin. This bacterial retinal-protein complex resembles the visual pigment and, after the absorption of protons, translocates H(+) across the cell membrane, leading to an electrochemical gradient for protons between the inside and the outside of the cell. Thus, light is an alternate source of energy in these bacteria, in addition to terminal oxidation. The paper deals with work on light-driven transport in H. halobium with cell envelope vesicles. The discussion covers light-driven movements of H(+), Na(+), and K(+); light-driven amino acid transport; and apparent allosteric control of amino acid transport. The scheme of energy coupling in H. halobium vesicles appears simple, its quantitative details are quite complex and reveal regulatory phenomena. More knowledge is required of the way the coupling components are regulated by the ion gradients present.

  3. Excitation signal processing times in Halobacterium halobium phototaxis.


    Phototaxis responses of Halobacterium halobium were monitored with a computerized cell-tracking system coupled to an electronic shutter controlling delivery of photostimuli. Automated analysis of rates of change in direction and linear speeds provided detection of swimming reversals with 67 ms resolution, permitting measurement of distinct phases of the responses to attractant and repellent stimuli. After stimulation, there was a latency period in which the population reversal frequency was u...

  4. Buffering capacity and membrane H+ conductance of Halobacterium halobium.

    Rius, N; Lorén, J G


    Buffering capacity and membrane H+ conductance were measured in Halobacterium halobium suspensions in the light and in the dark over a wide range of external pH. The values of both variables for this archaeobacterium were significantly higher than those found for eubacteria in other reports. It appears from our results that the special chemical composition of the cell envelope and the movement of ions, mainly protons, may influence the magnitude of the buffering power and the H+ membrane conductance of these cells.

  5. Excitation signal processing times in Halobacterium halobium phototaxis.

    Sundberg, S A; Alam, M; Spudich, J L


    Phototaxis responses of Halobacterium halobium were monitored with a computerized cell-tracking system coupled to an electronic shutter controlling delivery of photostimuli. Automated analysis of rates of change in direction and linear speeds provided detection of swimming reversals with 67 ms resolution, permitting measurement of distinct phases of the responses to attractant and repellent stimuli. After stimulation, there was a latency period in which the population reversal frequency was unchanged, followed by an excitation phase in which reversal frequency increased, and a slower adaptation phase in which reversal frequency returned to its prestimulus value. A step-decrease in illumination of the attractant receptor slow-cycling or sensory rhodopsin (SR) (lambda max, 587 nm) was interpreted by the cells as an unfavorable stimulus and, after a minimum latency of 0.70 +/- 0.14 s, induced swimming reversals with the peak response occurring 1.34 +/- 0.07 s after onset of the stimulus. Two distinct repellent responses in the near UV/blue were observed. One was a reversal response to 400 nm light, which was dependent on orange-red background illumination as expected for the photointermediate repellent form of SR (lambda max, 373 nm). The minimum latency of this response was approximately the same as that of the SR attractant system. The second was a reversal response with shorter minimum latency (0.40 +/- 0.07 s) to light of longer wavelength (450 nm) than absorbed by the known SR repellent form. This result confirms recent findings of an additional repellent photosystem in this spectral range. Further, the longer wavelength repellent response is independent of orange-red background illumination, indicating that the photoreceptor mediating this response is not a photointermediate of SR.

  6. Resonance Raman Spectroscopy of Purple Membrane from Halobacterium Halobium.

    Argade, Pramod Vasant

    Purple membrane from the halophilic bacteria, Halobacterium halobium, contains the protein, bacteriorhodopsin, which functions as a light transducing proton pump. Understanding the molecular mechanism underlying the functioning of bacteriorhodopsin is a key problem in membrane biophysics. After absorbing a photon, this protein cycles through a series of characteristic intermeidate states and pumps H('+) ions across the membrane. In this way, the energy of the absorbed photon is stored in the electrochemical potential gradient formed across the membrane. This energy is subsequently available for metabolism by the bacterium. Bacteriorhodopsin consists of a retinal chromophore (which is responsible for the purple color) bound to the protein, bacterioopsin, whose sequence is known and consists of 248 amino acid residues. There is evidence that conformational changes in the chromophore may contribute to the proton pumping action. Resonance Raman light scattering provides a selective tool to monitor the conformational changes in the chromophore during the proton pumping cycle. This dissertation consists of applying resonance Raman light scattering in conjunction with a variety of newly developed experimental techniques to gain information about the mode of action of bacteriorhodopsin. By selective isotopic labelling of (epsilon)-amino nitrogen of the lysine residues of the protein, the site of attachment of the chromophore with the protein was verified by in situ measurements. Also, a model proposing a secondary interaction of the chromophore with a lysine residue other than the binding site of the chromophore was tested using this method. Furthermore, by selective isotopic labelling of only a part of the protein the location of the lysine on the protein to which the chromophore is bound, was found by in situ measurements to be the fragment consisting of amino acid residues 72 through 248 of the protein. This is inconsistent with the previously reported binding site at

  7. Halorhodopsin and photosensory behaviour in Halobacterium halobium mutant strain L-33

    Traulich, B.; Wagner, G. (Botanisches Institut l der Justus-Liebig-Universitat, Giessen (Germany, F.R.)); Hildebrand, E.; Schimz, A. (Kernforschungsanlage Juelich G.m.b.H. (Germany, F.R.). Inst. fuer Neurobiologie); Lanyi, J.K. (California Univ., Irvine (USA))


    Halobacterium halobium, strain L-33, which is deficient in bacteriorhodopsin (BR) but synthesizes increased amounts of halorhodopsin (HR), responds to changes in fluence rate with visible light or with UV light. The observations support an earlier report that BR is not essential for photosensing in H. halobium. In the UV-range, changes in light intensity elicit the maximal response at lambda = 370 nm. In the visible range, changes in light intensity show the maximal response at lambda = 565 nm and a secondary peak at lambda = 590 nm. The latter corresponds to the absorption maximum of HR (lambdasub(max) = 588 nm). This light-energy converting retinal pigment of H. halobium thus appears to contribute to photosensory behavior.

  8. Role of Protein Methylation in Halobacterium halobium Phototaxis


    suiscnentemlclrbasis of sensory ’and. energy transduction by archaebacterial rhodopsins, phototnnsducerS In Halobacterium hoiobium.. Two of these, rhodopsin-liki...and SR-Il, Ajj 487 am), regulate the cells’~ swimming behavior, enabling thern to migrate into preferred regions of light intensity and color ...bacterial motility, photosensory receptors, retinal, color -sensing, photadetection, spectroscopy 19. will be used for preparation of in oligonucleotide

  9. Light-activated amino acid transport in Halobacterium halobium envelope vesicles

    Macdonald, R. E.; Lanyi, J. K.


    Vesicles prepared from Halobacterium halobium cell envelopes accumulate amino acids in response to light-induced electrical and chemical gradients. Nineteen of 20 commonly occurring amino acids have been shown to be actively accumulated by these vesicles in response to illumination or in response to an artificially created Na+ gradient. On the basis of shared common carriers the transport systems can be divided into eight classes, each responsible for the transport of one or several amino acids: arginine, lysine, histidine; asparagine, glutamine; alanine, glycine, threonine, serine; leucine, valine, isoleucine, methionine; phenylalanine, tyrosine, tryptophan; aspartate; glutamate; proline. Available evidence suggests that these carriers are symmetrical in that amino acids can be transported equally well in both directions across the vesicle membranes. A tentative working model to account for these observations is presented.

  10. Microcalorimetric Studies on Gene Promoter Function of Cloned DNA Fragements from Halobacterium halobium J7 Plasmid pHH205 in Escherichia coli TG1

    LEI,Ke-Lin; HOU,Han-Na; LIU,Yi; YE,Xue-Cheng; SHEN,Ping


    Halobacterium halobium is a typical kind of extremely halophilic bacterium. Combined with the antibiotic resistance assay, the microcalorimetric method was used to study the promoter function of the cloned DNA fragments from Halobacterium halobium J7 plasmid pHH205 in Escherichia coli TG1. The promoter probe vector, plasmid pKK232-8, was used to form the recombinants. The DNA fragment, which is the promoter for the chloramphenicol acetyl transferase (CAT) gene in plasmid pKK232-8, is about 800 bp, and the chloramphenicol resistance level presented by IC50 is about 200 μg·mL-1, which suggests a high promoter activity. The conclusions show that there probably exist double-function or trinary-function gene promoters in Halobacterium halobium, and Archaea may contain rich genetic resources.

  11. Photosensory behaviour of a bacteriorhodopsin-deficient mutant, ET-15, of Halobacterium halobium

    Hildebrand, E.; Schimz, A. (Kernforschungsanlage Juelich G.m.b.H. (Germany, F.R.). Inst. fuer Neurobiologie)


    Halobacterium halobium, strain ET-15, which does not contain detectable amounts of bacteriorhodopsin (BR) shows behavioral responses to UV and yellow-green light. Attractant stimuli, i.e. light-increases in the yellow-green range or light-decreases in the UV, suppress the spontaneous reversals of the swimming direction for a certain time. Repellent stimuli, i.e. light-decreases in the yellow-green range or light-increases in the UV, elicit an additional reversal response after a few seconds. Action spectra of both sensory photosystems, PS 370 and PS 565, were measured with attractant as well as with repellent stimuli. As in BR-containing cells, maximal sensitivity was always found at 370 nm for the UV-system and at 565 nm for the long-wavelength system. Fluence-response curves at 370 and 565 nm obtained with strain ET-15 and with a BR-containing strain show that the sensitivity of both photosystems is not reduced in the absence of BR. It is concluded that BR is required neither for PS 565 nor for PS 370. Instead retinal-containing pigments different from BR have to be assumed to mediate photosensory behavior.

  12. Identification of a third rhodopsin-like pigment in phototactic Halobacterium halobium.

    Bogomolni, R A; Spudich, J L


    Mutant Halobacterium halobium strains deficient in all previously reported rhodopsin-like pigments show phototaxis responses comparable to those of wild-type strains. Spectroscopic analysis reveals the presence of a third retinal-containing pigment in the cells and their membrane fractions. It undergoes a photoreaction cycle with a half-time of approximately equal to 1 sec at room temperature and at physiological light intensities the photostationary state of the pigment consists of two species, one absorbing in the 580- to 590-nm region and the other at 373 nm, both of which are photoactive. Illumination of the long-wavelength species generates the 373-nm intermediate, which upon photoexcitation reconverts to the long-wavelength form. Therefore, changes in the relative light intensities in the long- and short-wavelength regions of the visible spectrum cause opposing shifts in the photostationary state. The spectral sensitivity of this pigment correlates with the color-discriminating phototaxis sensitivities of this organism and strongly suggests that it is the sensory photoreceptor.

  13. Microarray analysis in the archaeon Halobacterium salinarum strain R1.

    Jens Twellmeyer

    Full Text Available BACKGROUND: Phototrophy of the extremely halophilic archaeon Halobacterium salinarum was explored for decades. The research was mainly focused on the expression of bacteriorhodopsin and its functional properties. In contrast, less is known about genome wide transcriptional changes and their impact on the physiological adaptation to phototrophy. The tool of choice to record transcriptional profiles is the DNA microarray technique. However, the technique is still rarely used for transcriptome analysis in archaea. METHODOLOGY/PRINCIPAL FINDINGS: We developed a whole-genome DNA microarray based on our sequence data of the Hbt. salinarum strain R1 genome. The potential of our tool is exemplified by the comparison of cells growing under aerobic and phototrophic conditions, respectively. We processed the raw fluorescence data by several stringent filtering steps and a subsequent MAANOVA analysis. The study revealed a lot of transcriptional differences between the two cell states. We found that the transcriptional changes were relatively weak, though significant. Finally, the DNA microarray data were independently verified by a real-time PCR analysis. CONCLUSION/SIGNIFICANCE: This is the first DNA microarray analysis of Hbt. salinarum cells that were actually grown under phototrophic conditions. By comparing the transcriptomics data with current knowledge we could show that our DNA microarray tool is well applicable for transcriptome analysis in the extremely halophilic archaeon Hbt. salinarum. The reliability of our tool is based on both the high-quality array of DNA probes and the stringent data handling including MAANOVA analysis. Among the regulated genes more than 50% had unknown functions. This underlines the fact that haloarchaeal phototrophy is still far away from being completely understood. Hence, the data recorded in this study will be subject to future systems biology analysis.

  14. The Improved Method for Isolation of Photochrome Trans-membrane Protein Bacteriorhodopsin from Purple Membranes of Halobacterium Halobacterium Halobium ET 1001

    Oleg Mosin


    Full Text Available It was developed the improved method for isolation of photochrome trans-membraine protein bacteriorhodopsin (output – 5 mg from 100 g of wet biomass capable to transform light energy to electrochemical energy of generated protons H+ and АТP. The protein was isolated from purple membranes of photo-organotrophic halobacterium Halobacterium halobium ET 1001 by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weight molecular impurities, cellular RNA, carotenoids and lipids, solubilization with 0.5% (w/v SDS-Na, fractionation by MeOH and column gel permeation chromatography (GPC of the final protein on Sephadex G-200 with 0.1% (w/v SDS-Na and 2.5 mM ETDA. The homogeneity of the isolated bacteriorhodopsin was proved by combination of preparative and analytical methods, including elecrtophoresis in 12.5% (w/v PAAG with 0.1% (w/v SDS-Na and regeneration of apomembranes with 13-trans-retinal.

  15. Continuous hydrogen evolution by an immobilized combined system of Phormidium valderianum, Halobacterium halobium and Escherichia coli in a packed bed reactor

    Patel, S.; Madamwar, D. [Sardar Patel Univ., Vallabh Vidyanagar (India)


    Long-term hydrogen evolution (11 days) was obtained when Phormidium valderianum was coupled with a mixture of halophilic Halobacterium halobium and salt tolerant Escherichia coli under a cyclic illumination on/off cycle of 18 h dark and 6 h light in an immobilized packed bed reactor. Various conditions such as flow rate, temperature and pH have been optimized for maximum continuous evolution. (author)

  16. Further characterization of particulate fractions from lysed cell envelopes of Halobacterium halobium and isolation of gas vacuole membranes.

    Toeckenius, W; Kunau, W H


    Lysates of cell envelopes from Halobacterium halobium have been separated into four fractions. A soluble, colorless fraction (I) containing protein, hexosamines, and no lipid is apparently derived from the cell wall. A red fraction (II), containing approximately 40 per cent lipid, 60 per cent protein, and a small amount of hexosamines consists of cell membrane disaggregated into fragments of small size. A third fraction (III) of purple color consists of large membrane sheets and has a very similar composition to II, containing the same classes of lipids but no hexosamines; its buoyant density is 1.18 g/ml. The fourth fraction (IV) has a buoyant density of 1.23 g/ml and contains the "intracytoplasmic membranes." These consist mainly of protein, and no lipid can be extracted with chloroform-methanol. Fractions I and II, which result from disaggregation of cell wall and cell membrane during lysis, contain a high proportion of dicarboxyl amino acids; this is in good agreement with the assumption that disruption of the cell envelope upon removal of salt is due to the high charge density. The intracytoplasmic membranes (IV) represent the gas vacuole membranes in the collapsed state. In a number of mutants that have lost the ability to form gas vacuoles, no vacuole membranes or any structure that could be related to them has been found.

  17. Transcriptional control by two leucine-responsive regulatory proteins in Halobacterium salinarum R1

    Tarasov Valery


    Full Text Available Abstract Background Archaea combine bacterial-as well as eukaryotic-like features to regulate cellular processes. Halobacterium salinarum R1 encodes eight leucine-responsive regulatory protein (Lrp-homologues. The function of two of them, Irp (OE3923F and lrpA1 (OE2621R, were analyzed by gene deletion and overexpression, including genome scale impacts using microarrays. Results It was shown that Lrp affects the transcription of multiple target genes, including those encoding enzymes involved in amino acid synthesis, central metabolism, transport processes and other regulators of transcription. In contrast, LrpA1 regulates transcription in a more specific manner. The aspB3 gene, coding for an aspartate transaminase, was repressed by LrpA1 in the presence of L-aspartate. Analytical DNA-affinity chromatography was adapted to high salt, and demonstrated binding of LrpA1 to its own promoter, as well as L-aspartate dependent binding to the aspB3 promoter. Conclusion The gene expression profiles of two archaeal Lrp-homologues report in detail their role in H. salinarum R1. LrpA1 and Lrp show similar functions to those already described in bacteria, but in addition they play a key role in regulatory networks, such as controlling the transcription of other regulators. In a more detailed analysis ligand dependent binding of LrpA1 was demonstrated to its target gene aspB3.

  18. Novel pili-like surface structures of Halobacterium salinarum strain R1 are crucial for surface adhesion

    Gerald eLosensky


    Full Text Available It was recently shown that haloarchaeal strains of different genera are able to adhere to surfaces and form surface-attached biofilms. However the surface structures mediating the adhesion were still unknown. We have identified a novel surface structure with Halobacterium salinarum strain R1, crucial for surface adhesion. Electron microscopic studies of surface-attached cells frequently showed pili-like surface structures of two different diameters that were irregularly distributed on the surface. The thinner filaments, 7 - 8 nm in diameter, represented a so far unobserved novel pili-like structure. Examination of the Hbt. salinarum R1 genome identified two putative gene loci (pil-1 and pil-2 encoding type IV pilus biogenesis complexes besides the archaellum encoding fla gene locus. Both pil-1 and pil-2 were expressed as transcriptional units, and the transcriptional start of pil-1 was identified. In silico analyses revealed that the pil-1 locus is present with other euryarchaeal genomes whereas the pil-2 is restricted to haloarchaea. Comparative real time qRT-PCR studies indicated that the general transcriptional activity was reduced in adherent versus planktonic cells. In contrast, the transcription of pilB1 and pilB2, encoding putative type IV pilus assembly ATPases, was induced in comparison to the archaella assembly/motor ATPase (flaI and the ferredoxin gene. Mutant strains were constructed that incurred a flaI deletion or flaI/pilB1 gene deletions. The absence of flaI caused the loss of the archaella while the additional absence of pilB1 led to loss of the novel pili-like surface structures. The ΔflaI/ΔpilB1 double mutants showed a 10-fold reduction in surface adhesion compared to the parental strain. Since surface adhesion was not reduced with the non-archaellated ΔflaI mutants, the pil-1 filaments have a distinct function in the adhesion process.

  19. 盐生盐杆菌RM07 DNA片段在大肠杆菌中的定点诱变和启动子功能分析%Site-directed Mutagenesis and Promoter Functional Analysis of RM07 DNA Fragment from Halobacterium halobium in Escherichia coli

    杨洋; 沈萍


    将来源于嗜盐古生菌--盐生盐杆菌(Halobacterium halobium)基因组的RM07 DNA片段以正反两个方向分别插人大肠杆菌启动子探针载体pKK232-8携带的报告基因--氯霉素抗性基因(cat)的上游,得到RM07-cat融合的质粒pRM07-1(+)和pRM07-1(-),将其分别转入大肠杆菌HB101,进而检测了不同转化子菌株的氯霉素抗性水平和细胞内氯霉素乙酰转移酶蛋白质浓度.结果表明:正向的RM07片段在真细菌(大肠杆菌)中具有启动子活性,能够驱动cat报告基因的表达;而反向的RM07片段在大肠杆菌中不具有启动子活性.对RM07片段进行了定点诱变分析,检测了特定核苷酸突变对启动子活性的影响,结果进一步精确定位了RM07片段中对在大肠杆菌中的启动子功能有重要作用的关键碱基,并且通过改造RM07片段的碱基组成成分大幅提高了其在大肠杆菌中的启动子活性.

  20. Shuttle vectors for the archaebacterium Halobacterium volcanii.

    Lam, W. L.; Doolittle, W F


    Progress in archaebacterial molecular biology requires tools for genetic analysis. We describe vectors that can be selected and maintained in either Halobacterium volcanii or Escherichia coli. A genetic determinant for resistance to the 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor mevinolin was isolated by "shotgun cloning" into a derivative of the endogenous H. volcanii plasmid pHV2, to form pWL2, which transforms sensitive H. volcanii to mevinolin resistance at high frequency. ...

  1. Deciphering a pathway of Halobacterium salinarum N-glycosylation

    Kandiba, Lina; Eichler, Jerry


    Genomic analysis points to N-glycosylation as being a common posttranslational modification in Archaea. To date, however, pathways of archaeal N-glycosylation have only been described for few species. With this in mind, the similarities of N-linked glycans decorating glycoproteins in the haloarchaea Haloferax volcanii and Halobacterium salinarum directed a series of bioinformatics, genetic, and biochemical experiments designed to describe that Hbt. salinarum pathway responsible for biogenesis...

  2. Halobacterium denitrificans sp. nov., an extremely halophilic denitrifying bacterium

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.


    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  3. Halobacterium denitrificans sp. nov. - An extremely halophilic denitrifying bacterium

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.


    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  4. Model Construction and Analysis of Respiration in Halobacterium salinarum.

    Cherryl O Talaue

    Full Text Available The archaeon Halobacterium salinarum can produce energy using three different processes, namely photosynthesis, oxidative phosphorylation and fermentation of arginine, and is thus a model organism in bioenergetics. Compared to its bacteriorhodopsin-driven photosynthesis, less attention has been devoted to modeling its respiratory pathway. We created a system of ordinary differential equations that models its oxidative phosphorylation. The model consists of the electron transport chain, the ATP synthase, the potassium uniport and the sodium-proton antiport. By fitting the model parameters to experimental data, we show that the model can explain data on proton motive force generation, ATP production, and the charge balancing of ions between the sodium-proton antiporter and the potassium uniport. We performed sensitivity analysis of the model parameters to determine how the model will respond to perturbations in parameter values. The model and the parameters we derived provide a resource that can be used for analytical studies of the bioenergetics of H. salinarum.

  5. Deciphering a pathway of Halobacterium salinarum N-glycosylation.

    Kandiba, Lina; Eichler, Jerry


    Genomic analysis points to N-glycosylation as being a common posttranslational modification in Archaea. To date, however, pathways of archaeal N-glycosylation have only been described for few species. With this in mind, the similarities of N-linked glycans decorating glycoproteins in the haloarchaea Haloferax volcanii and Halobacterium salinarum directed a series of bioinformatics, genetic, and biochemical experiments designed to describe that Hbt. salinarum pathway responsible for biogenesis of one of the two N-linked oligosaccharides described in this species. As in Hfx. volcanii, where agl (archaeal glycosylation) genes that encode proteins responsible for the assembly and attachment of a pentasaccharide to target protein Asn residues are clustered in the genome, Hbt. salinarum also contains a group of clustered homologous genes (VNG1048G-VNG1068G). Introduction of these Hbt. salinarum genes into Hfx. volcanii mutant strains deleted of the homologous sequence restored the lost activity. Moreover, transcription of the Hbt. salinarum genes in the native host, as well as in vitro biochemical confirmation of the predicted functions of several of the products of these genes provided further support for assignments made following bioinformatics and genetic experiments. Based on the results obtained in this study, the first description of an N-glycosylation pathway in Hbt. salinarum is offered.

  6. Regulatory Multidimensionality of Gas Vesicle Biogenesis in Halobacterium salinarum NRC-1

    Andrew I. Yao


    Full Text Available It is becoming clear that the regulation of gas vesicle biogenesis in Halobacterium salinarum NRC-1 is multifaceted and appears to integrate environmental and metabolic cues at both the transcriptional and posttranscriptional levels. The mechanistic details underlying this process, however, remain unclear. In this manuscript, we quantify the contribution of light scattering made by both intracellular and released gas vesicles isolated from Halobacterium salinarum NRC-1, demonstrating that each form can lead to distinct features in growth curves determined by optical density measured at 600 nm (OD600. In the course of the study, we also demonstrate the sensitivity of gas vesicle accumulation in Halobacterium salinarum NRC-1 on small differences in growth conditions and reevaluate published works in the context of our results to present a hypothesis regarding the roles of the general transcription factor tbpD and the TCA cycle enzyme aconitase on the regulation of gas vesicle biogenesis.

  7. [R1 resection of esophageal carcinoma].

    Gockel, I; Wittekind, C


    The microscopic identification of residual tumor tissue in the oral or aboral resection margins (R1 resection) of esophageal specimens following oncologic esophageal resection, increases the risk of tumor recurrence and disease-related morbidity. Esophageal resection with its associated risks is only meaningful, if an R0 situation can be safely achieved. The relevance of microscopic involvement of the circumferential resection margin (CRM) in esophageal carcinoma in its different definitions by the British and the American Societies of Pathology has up to now never been investigated in a prospective study. According to the German S3 guideline, radiochemotherapy should be performed in a postoperatively proven R1 situation, which cannot be converted by a curative extended re-resection into an R0 situation or in unfavorable conditions for an extended re-resection, independent of neoadjuvant therapy. In the case of an R1 situation in the region of the CRM, an extended re-resection is not simply possible on account of the anatomical conditions with corresponding limitations by the aorta and the spinal column, in contrast to extensions of the re-resection orally or aborally.

  8. Comparison of two extreme halophilic Halobacterium noricense strains on DNA and protein level

    Bader, Miriam; Cherkouk, Andrea [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). HZDR Young Investigator Group; Flemming, Katrin [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). Biogeochemistry; Swanson, J.S. [Los Alamos National Laboratory, Carlsbad, NM (United States)


    Two strains of the halophilic archaeon Halobacterium noricense isolated from rock salt of different locations were used for interaction studies with uranium. It was found that both strains showed similar, atypical bioassociation kinetics accompanied by cell agglomeration as a stress response. The 16S rRNA gene sequences of both strains had a high similarity (> 99 %). However, differences in the whole protein pattern were apparent.

  9. Novel benzimidazole-based MCH R1 antagonists.

    Carpenter, Andrew J; Al-Barazanji, Kamal A; Barvian, Kevin K; Bishop, Michael J; Britt, Christy S; Cooper, Joel P; Goetz, Aaron S; Grizzle, Mary K; Hertzog, Donald L; Ignar, Diane M; Morgan, Ronda O; Peckham, Gregory E; Speake, Jason D; Swain, Will R


    The identification of an MCH R1 antagonist screening hit led to the optimization of a class of benzimidazole-based MCH R1 antagonists. Structure-activity relationships and efforts to optimize pharmacokinetic properties are detailed along with the demonstration of the effectiveness of an MCH R1 antagonist in an animal model of obesity.

  10. Resistance of the Extreme Halophile Halobacterium sp. NRC-1 to Multiple Stresses

    Gygli, Patrick E.; Prajapati, Surendra; DeVeaux, Linda C.; DasSarma, Shiladitya; DasSarma, Priya; Mestari, Mohammed Amine; Wells, Douglas P.


    The model Archaeon Halobacterium sp. NRC-1 is an extreme halophile known for its resistance to multiple stressors, including electron-beam and ultraviolet radiation. It is a well-developed system with a completely sequenced genome and extensive post-genomic tools for the study of a variety of biological processes. To further understand the mechanisms of Halobacterium's, radiation resistance, we previously reported the selection for multiple independent highly resistant mutants using repeated exposure to high doses of 18-20 MeV electrons using a medical S-band Linac. Molecular analysis of the transcriptional profile of several of these mutants revealed a single common change: upregulation of the rfa3 operon. These genes encode proteins homologous to the subunits of eukaryotic Replication Protein A (RPA), a DNA binding protein with major roles in DNA replication, recombination, and repair. This operon has also been implicated in a somewhat lesser role in resistance of wild type Halobacterium to ultraviolet radiation, suggesting common mechanisms for resistance. To further understand the mechanism of radiation resistance in the mutant strains, we measured the survival after exposure to both electron-beam and ultraviolet radiation, UV-A, B, and C All mutant strains showed increased resistance to electrons when compared with the parent. However, the mutant strains do not display increased UV resistance, and in one case is more sensitive than the parent strain. Thus, the protective role of increased RPA expression within a cell may be specific to the DNA damage caused by the different physical effects induced by high energy electron-beam radiation.

  11. Salt-Dependent Conditional Protein Splicing of an Intein from Halobacterium salinarum.

    Reitter, Julie N; Cousin, Christopher E; Nicastri, Michael C; Jaramillo, Mario V; Mills, Kenneth V


    An intein from Halobacterium salinarum can be isolated as an unspliced precursor protein with exogenous exteins after Escherichia coli overexpression. The intein promotes protein splicing and uncoupled N-terminal cleavage in vitro, conditional on incubation with NaCl or KCl at concentrations of >1.5 M. The protein splicing reaction also is conditional on reduction of a disulfide bond between two active site cysteines. Conditional protein splicing under these relatively mild conditions may lead to advances in intein-based biotechnology applications and hints at the possibility that this H. salinarum intein could serve as a switch to control extein activity under physiologically relevant conditions.

  12. Genomic perspective on the photobiology of Halobacterium species NRC-1, a phototrophic, phototactic, and UV-tolerant haloarchaeon.

    Dassarma, S; Kennedy, S P; Berquist, B; Victor Ng, W; Baliga, N S; Spudich, J L; Krebs, M P; Eisen, J A; Johnson, C H; Hood, L


    Halobacterium species display a variety of responses to light, including phototrophic growth, phototactic behavior, and photoprotective mechanisms. The complete genome sequence of Halobacterium species NRC-1 (Proc Natl Acad Sci USA 97: 12176-12181, 2000), coupled with the availability of a battery of methods for its analysis makes this an ideal model system for studying photobiology among the archaea. Here, we review: (1) the structure of the 2.57 Mbp Halobacterium NRC-1 genome, including a large chromosome, two minichromosomes, and 91 transposable IS elements; (2) the purple membrane regulon, which programs the accumulation of large quantities of the light-driven proton pump, bacteriorhodopsin, and allows for a period of phototrophic growth; (3) components of the sophisticated pathways for color-sensitive phototaxis; (4) the gas vesicle gene cluster, which codes for cell buoyancy organelles; (5) pathways for the production of carotenoid pigments and retinal, (6) processes for the repair of DNA damage; and (7) putative homologs of circadian rhythm regulators. We conclude with a discussion of the power of systems biology for comprehensive understanding of Halobacterium NRC-1 photobiology.

  13. Association of Eu(III) and Cm(III) with Bacillus subtilis and Halobacterium salinarum

    Ozaki, Takuo; Kimura, Takaumi; Ohnuki, Toshihiko; Yoshida, Zenko [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment; Gillow, Jeffrey B.; Francis, Arokiasamy J. [Brookhaven National Laboratory, Upton, NY (United States)


    Adsorption behavior of Eu(III) and Cm(III) by Bacillus subtilis and Halobacterium salinarum was investigated. Both microorganisms showed almost identical pH dependence on the distribution ratio (K{sub d}) of the metals examined, i.e., K{sub d} of Eu(III) and Cm(III) increased with an increase of pH. The coordination state of Eu(III) adsorbed on the microorganisms was studied by time-resolved laser-induced fluorescence spectroscopy (TRLFS). The coordination states of Eu(III) adsorbed on the B. subtilis and H. salinarum was of different characteristics. H. salinarum exhibited more outer-spherical interaction with Eu(III) than B. subtilis. (author)

  14. Sequence of the 16S Ribosomal RNA from Halobacterium volcanii, an Archaebacterium.

    Gupta, R; Lanter, J M; Woese, C R


    The sequence of the 16S ribosomal RNA (rRNA) from the archaebacterium Halobacterium volcanii has been determined by DNA sequencing methods. The archaebacterial rRNA is similar to its eubacterial counterpart in secondary structure. Although it is closer in sequence to the eubacterial 16S rRNA than to the eukaryotic 16S-like rRNA, the H. volcanii sequence also shows certain points of specific similarity to its eukaryotic counterpart. Since the H. volcanii sequence is closer to both the eubacterial and the eukaryotic sequences than these two are to one another, it follows that the archaebacterial sequence resembles their common ancestral sequence more closely than does either of the other two versions.

  15. Oracle BAM 11gR1 Handbook

    Wang, Pete


    "Oracle BAM 11gR1 Handbook" is a practical best practices tutorial focused entirely on Oracle Business Activity Monitoring. An intermediate-to-advanced guide, step-by-step instructions and an accompanying demo project will help SOA report developers through application development and producing dashboards and reports. If you are a developer/report developer or SOA Architect who wants to learn valuable Oracle BAM best practices for monitoring your operations in real time, then "Oracle BAM 11gR1 Handbook" is for you. Administrators will also find the book useful. You should already be comfortabl

  16. Analytical properties of the $R^{1/m}$ law

    Ciotti, L


    In this paper we describe some analytical properties of the R^{1/m} law proposed by Sersic to categorize the photometric profiles of elliptical galaxies. In particular, we present the full asymptotic expansion for the dimensionless scale factor b(m) that is introduced when referring the profile to the standard effective radius. Surprisingly, our asymptotic analysis turns out to be useful even for values of m as low as unity, thus providing a unified analytical tool for observational and theoretical investigations based on the R^{1/m} law for the entire range of interesting photometric profiles, from spiral to elliptical galaxies.

  17. Analytical properties of the $R^{1/m}$ luminosity law

    Ciotti, L


    In this paper we describe some analytical properties of the R^{1/m} law proposed by Sersic (1968) to categorize the photometric profiles of elliptical galaxies. In particular, we present the full asymptotic expansion for the dimensionless scale factor b(m) that is introduced when referring the profile to the standard effective radius. Surprisingly, our asymptotic analysis turns out to be useful even for values of m as low as unity, thus providing a unified analytical tool for observational and theoretical investigations based on the R^{1/m} law for the entire range of interesting photometric profiles, from spiral to elliptical galaxies.

  18. The cobY gene of the archaeon Halobacterium sp. strain NRC-1 is required for de novo cobamide synthesis.

    Woodson, J D; Peck, R F; Krebs, M P; Escalante-Semerena, J C


    Genetic and nutritional analyses of mutants of the extremely halophilic archaeon Halobacterium sp. strain NRC-1 showed that open reading frame (ORF) Vng1581C encodes a protein with nucleoside triphosphate:adenosylcobinamide-phosphate nucleotidyltransferase enzyme activity. This activity was previously associated with the cobY gene of the methanogenic archaeon Methanobacterium thermoautotrophicum strain DeltaH, but no evidence was obtained to demonstrate the direct involvement of this protein in cobamide biosynthesis in archaea. Computer analysis of the Halobacterium sp. strain NRC-1 ORF Vng1581C gene and the cobY gene of M. thermoautotrophicum strain DeltaH showed the primary amino acid sequence of the proteins encoded by these two genes to be 35% identical and 48% similar. A strain of Halobacterium sp. strain NRC-1 carrying a null allele of the cobY gene was auxotrophic for cobinamide-GDP, a known intermediate of the late steps of cobamide biosynthesis. The auxotrophic requirement for cobinamide-GDP was corrected when a wild-type allele of cobY was introduced into the mutant strain, demonstrating that the lack of cobY function was solely responsible for the observed block in cobamide biosynthesis in this archaeon. The data also show that Halobacterium sp. strain NRC-1 possesses a high-affinity transport system for corrinoids and that this archaeon can synthesize cobamides de novo under aerobic growth conditions. To the best of our knowledge this is the first genetic and nutritional analysis of cobalamin biosynthetic mutants in archaea.

  19. Poorly conserved ORFs in the genome of the archaea Halobacterium sp. NRC-1 correspond to expressed proteins.

    Shmuely, H; Dinitz, E; Dahan, I; Eichler, J; Fischer, D; Shaanan, B


    A large fraction of open reading frames (ORFs) identified as 'hypothetical' proteins correspond to either 'conserved hypothetical' proteins, representing sequences homologous to ORFs of unknown function from other organisms, or to hypothetical proteins lacking any significant sequence similarity to other ORFs in the databases. Elucidating the functions and three-dimensional structures of such orphan ORFs, termed ORFans or poorly conserved ORFs (PCOs), is essential for understanding biodiversity. However, it has been claimed that many ORFans may not encode for expressed proteins. A genome-wide experimental study of 'paralogous PCOs' in the halophilic archaea Halobacterium sp. NRC-1 was conducted. Paralogous PCOs are ORFs with at least one homolog in the same organism, but with no clear homologs in other organisms. The results reveal that mRNA is synthesized for a majority of the Halobacterium sp. NRC-1 paralogous PCO families, including those comprising relatively short proteins, strongly suggesting that these Halobacterium sp. NRC-1 paralogous PCOs correspond to true, expressed proteins. Hence, further computational and experimental studies aimed at characterizing PCOs in this and other organisms are merited. Such efforts could shed light on PCOs' functions and origins, thereby serving to elucidate the vast diversity observed in the genetic material.

  20. Essential and non-essential DNA replication genes in the model halophilic Archaeon, Halobacterium sp. NRC-1

    DasSarma Shiladitya


    Full Text Available Abstract Background Information transfer systems in Archaea, including many components of the DNA replication machinery, are similar to those found in eukaryotes. Functional assignments of archaeal DNA replication genes have been primarily based upon sequence homology and biochemical studies of replisome components, but few genetic studies have been conducted thus far. We have developed a tractable genetic system for knockout analysis of genes in the model halophilic archaeon, Halobacterium sp. NRC-1, and used it to determine which DNA replication genes are essential. Results Using a directed in-frame gene knockout method in Halobacterium sp. NRC-1, we examined nineteen genes predicted to be involved in DNA replication. Preliminary bioinformatic analysis of the large haloarchaeal Orc/Cdc6 family, related to eukaryotic Orc1 and Cdc6, showed five distinct clades of Orc/Cdc6 proteins conserved in all sequenced haloarchaea. Of ten orc/cdc6 genes in Halobacterium sp. NRC-1, only two were found to be essential, orc10, on the large chromosome, and orc2, on the minichromosome, pNRC200. Of the three replicative-type DNA polymerase genes, two were essential: the chromosomally encoded B family, polB1, and the chromosomally encoded euryarchaeal-specific D family, polD1/D2 (formerly called polA1/polA2 in the Halobacterium sp. NRC-1 genome sequence. The pNRC200-encoded B family polymerase, polB2, was non-essential. Accessory genes for DNA replication initiation and elongation factors, including the putative replicative helicase, mcm, the eukaryotic-type DNA primase, pri1/pri2, the DNA polymerase sliding clamp, pcn, and the flap endonuclease, rad2, were all essential. Targeted genes were classified as non-essential if knockouts were obtained and essential based on statistical analysis and/or by demonstrating the inability to isolate chromosomal knockouts except in the presence of a complementing plasmid copy of the gene. Conclusion The results showed that ten

  1. Genomic Analysis of the Extremely Halophilic Archaeon Halobacterium noricense CBA1132 Isolated from Solar Salt That Is an Essential Material for Fermented Foods.

    Lim, Seul Ki; Kim, Joon Yong; Song, Hye Seon; Kwon, Min-Sung; Lee, Jieun; Oh, Young Jun; Nam, Young-Do; Seo, Myung-Ji; Lee, Dong-Gi; Choi, Jong-Soon; Yoon, Changmann; Sohn, Eunju; Rahman, Md Arif-Ur; Roh, Seong Woon; Choi, Hak-Jong


    The extremely halophilic archaeon Halobacterium noricense is a member of the genus Halobacterium. Strain CBA1132 (= KCCM 43183, JCM 31150) was isolated from solar salt. The genome of strain CBA1132 assembled with 4 contigs, including three rRNA genes, 44 tRNA genes, and 3,208 open reading frames. Strain CBA1132 had nine putative CRISPRs and the genome contained genes encoding metal resistance determinants: copper-translocating P-type ATPase (CtpA), arsenical pump-driving ATPase (ArsA), arsenate reductase (ArsC), and arsenical resistance operon repressor (ArsR). Strain CBA1132 was related to Halobacterium noricense, with 99.2% 16S rRNA gene sequence similarity. Based on the comparative genomic analysis, strain CBA1132 has distinctly evolved; moreover, essential genes related to nitrogen metabolism were only detected in the genome of strain CBA1132 among the reported genomes in the genus Halobacterium. This genome sequence of Halobacterium noricense CBA1132 may be of use in future molecular biological studies.

  2. Main-Belt Comet P/2008 R1 (Garradd)

    Jewitt, David; Haghighipour, Nader


    We present a study of the newly-discovered main-belt comet P/2008 R1 (Garradd), an object with the dynamical characteristics of an asteroid and the physical characteristics of a comet. Photometry sets a limit to the effective radius of the nucleus at r_e < 0.7 km (red geometric albedo 0.05 assumed). The coma shows a secular fading in our data caused by the escape of dust particles from the near-nucleus environment. The optical reflection spectrum is a nearly neutral continuum devoid of gaseous emission lines, from which we derive a limit to the cyanide (CN) radical production rate of Q_CN <1.4e23/s and infer a mass loss rate <1.5 kg/s at the time of our observations. Unlike the first-reported main-belt comets, P/2008 R1 is not dynamically stable. The nearby 8:3 mean-motion resonance with Jupiter induces dynamical instability on timescales 20 to 30 Myr. Hence, we conclude that P/2008 R1 has recently arrived from a more stable source elsewhere. The high Tisserand parameter of the orbit (in fact, with T...

  3. Heterologous overexpression, purification and characterisation of an alcohol dehydrogenase (ADH2) from Halobacterium sp. NRC-1.

    Liliensiek, Ann-Kathrin; Cassidy, Jennifer; Gucciardo, Gabriele; Whitely, Cliadhna; Paradisi, Francesca


    Replacement of chemical steps with biocatalytic ones is becoming increasingly more interesting due to the remarkable catalytic properties of enzymes, such as their wide range of substrate specificities and variety of chemo-, stereo- and regioselective reactions. This study presents characterisation of an alcohol dehydrogenase (ADH) from the halophilic archaeum Halobacterium sp. NRC-1 (HsADH2). A hexahistidine-tagged recombinant version of HsADH2 (His-HsADH2) was heterologously overexpressed in Haloferax volcanii. The enzyme was purified in one step by immobilised Ni-affinity chromatography. His-HsADH2 was halophilic and mildly thermophilic with optimal activity for ethanol oxidation at 4 M KCl around 60 °C and pH 10.0. The enzyme was extremely stable, retaining 80 % activity after 30 days. His-HsADH2 showed preference for NADP(H) but interestingly retained 60 % activity towards NADH. The enzyme displayed broad substrate specificity, with maximum activity obtained for 1-propanol. The enzyme also accepted secondary alcohols such as 2-butanol and even 1-phenylethanol. In the reductive reaction, working conditions for His-HsADH2 were optimised for acetaldehyde and found to be 4 M KCl and pH 6.0. His-HsADH2 displayed intrinsic organic solvent tolerance, which is highly relevant for biotechnological applications.

  4. Nanoscale Electric Characteristics and Oriented Assembly of Halobacterium salinarum Membrane Revealed by Electric Force Microscopy

    Denghua Li


    Full Text Available Purple membranes (PM of the bacteria Halobacterium salinarum are a unique natural membrane where bacteriorhodopsin (BR can convert photon energy and pump protons. Elucidating the electronic properties of biomembranes is critical for revealing biological mechanisms and developing new devices. We report here the electric properties of PMs studied by using multi-functional electric force microscopy (EFM at the nanoscale. The topography, surface potential, and dielectric capacity of PMs were imaged and quantitatively measured in parallel. Two orientations of PMs were identified by EFM because of its high resolution in differentiating electrical characteristics. The extracellular (EC sides were more negative than the cytoplasmic (CP side by 8 mV. The direction of potential difference may facilitate movement of protons across the membrane and thus play important roles in proton pumping. Unlike the side-dependent surface potentials observed in PM, the EFM capacitive response was independent of the side and was measured to be at a dC/dz value of ~5.25 nF/m. Furthermore, by modification of PM with de novo peptides based on peptide-protein interaction, directional oriented PM assembly on silicon substrate was obtained for technical devices. This work develops a new method for studying membrane nanoelectronics and exploring the bioelectric application at the nanoscale.

  5. A transcription factor links growth rate and metabolism in the hypersaline adapted archaeon Halobacterium salinarum.

    Todor, Horia; Dulmage, Keely; Gillum, Nicholas; Bain, James R; Muehlbauer, Michael J; Schmid, Amy K


    Co-ordinating metabolism and growth is a key challenge for all organisms. Despite fluctuating environments, cells must produce the same metabolic outputs to thrive. The mechanisms underlying this 'growth homeostasis' are known in bacteria and eukaryotes, but remain unexplored in archaea. In the model archaeon Halobacterium salinarum, the transcription factor TrmB regulates enzyme-coding genes in diverse metabolic pathways in response to glucose. However, H. salinarum is thought not to catabolize glucose. To resolve this discrepancy, we demonstrate that TrmB regulates the gluconeogenic production of sugars incorporated into the cell surface S-layer glycoprotein. Additionally, we show that TrmB-DNA binding correlates with instantaneous growth rate, likely because S-layer glycosylation is proportional to growth. This suggests that TrmB transduces a growth rate signal to co-regulated metabolic pathways including amino acid, purine, and cobalamin biosynthesis. Remarkably, the topology and function of this growth homeostatic network appear conserved across domains despite extensive alterations in protein components.

  6. 4-Phenyldiazenyl-2-[(R-(1-phenylethyliminomethyl]phenol

    Yoshikazu Aritake


    Full Text Available The title chiral photochromic Schiff base compound, C21H19N3O, was synthesized from (R-1-phenylethylamine and the salicylaldehyde of an azobenzene derivative. The molecule corresponds to the phenol–imine tautomer, the C=N and N—C bond distances being 1.279 (3 and 1.477 (3 Å, respectively. An intramolecular O—H...N hydrogen bond occurs. The diazenyl group adopts a trans form with an N=N distance of 1.243 (3 Å.

  7. Electrical system regulations of the IEA-R1 reactor

    Mello, Jose Roberto de; Madi Filho, Tufic, E-mail:, E-mail: [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    The IEA-R1 reactor of the Nuclear and Energy Research Institute (IPEN-CNEN/SP), is a research reactor open pool type, designed and built by the U.S. firm Babcock and Wilcox, having, as coolant and moderator, deionized light water and beryllium and graphite, as reflectors. Until about 1988, the reactor safety systems received power from only one source of energy. As an example, it may be cited the control desk that was powered only by the vital electrical system 220V, which, in case the electricity fails, is powered by the generator group: no-break 220V. In the years 1989 and 1990, a reform of the electrical system upgrading to increase the reactor power and, also, to meet the technical standards of the ABNT (Associacao Brasileira de Normas Tecnicas) was carried out. This work has the objective of showing the relationship between the electric power system and the IEA-R1 reactor security. Also, it demonstrates that, should some electrical power interruption occur, during the reactor operation, this occurrence would not start an accident event. (author)

  8. Effects of cyclothiazide on GluR1/AMPA receptors

    Fucile, Sergio; Miledi, Ricardo; Eusebi, Fabrizio


    Cyclothiazide (CTZ), a positive allosteric modulator of ionotropic α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-type glutamate receptors, is used frequently to block the desensitization of both native and heterologously expressed AMPA receptors. Specifically, CTZ is known to produce a fast inhibition of AMPA receptor desensitization and a much slower potentiation of the AMPA current. By using patch-clamp techniques, the effects of CTZ were studied in HEK 293 cells stably transfected with the rat flip GluR1 subunit. Upon CTZ treatment, we found an increased apparent affinity for the agonist, a slow whole-cell current potentiation, a fast inhibition of desensitization, and a lengthening of single-channel openings. Furthermore, we show that CTZ alters the channel gating events modifying the relative contribution of different single-channel classes of conductance (γ), increasing and decreasing, respectively, the contributions of γM (medium) and γL (low) without altering that of the γH (high) conductance channels. We also present a kinetic model that predicts well all of the experimental findings of CTZ action. Finally, we suggest a protocol for standard cell treatment with CTZ to attain maximal efficacy of CTZ on GluR1 receptors. PMID:16473938

  9. (4R-4-[(1R-1,2-Dihydroxyethyl]-1-[(1R-1-phenylethyl]pyrrolidin-2-one

    Peter D. W. Boyd


    Full Text Available The title compound, C14H19NO3, was obtained as one of the two isomers of a Sharpless asymmetric dihydroxylation reaction of (1S-1-[(1R-1-phenylethyl]-4-vinylpyrrolidin-2-one. The absolute stereochemistry of this isomer was determined from the known stereochemistry (R at the bridge C atom between the phenyl and pyrrolidine rings. The molecules form one-dimensional tapes along the b axis via hydrogen bonding between the carbonyl O atom and the alcohol groups of neighbouring molecules. These assemble into sheets via interdigitative stacking of the phenyl rings and C—H...O interactions.

  10. ATP synthesis in Halobacterium saccharovorum: evidence that synthesis may be catalysed by an F0F1-ATP synthase

    Hochstein, L. I.


    Halobacterium saccharovorum synthesized ATP in response to a pH shift from 8 to 6.2. Synthesis was inhibited by carbonyl cyanide m-chloro-phenylhydrazone, dicyclohexylcarbodiimide, and azide. Nitrate, an inhibitor of the membrane-bound ATPase previously isolated from this organism, did not inhibit ATP synthesis. N-Ethymaleimide, which also inhibited this ATPase, stimulated the production of ATP. These observations suggested that H. saccharovorum synthesized and hydrolysed ATP using different enzymes and that the vacuolar-like ATPase activity previously described in H. saccharovorum was an ATPase whose function is yet to be identified.

  11. Molecular characterization of the Jatropha curcas JcR1MYB1 gene encoding a putative R1-MYB transcription factor

    Hui-Liang Li


    Full Text Available The cDNA encoding the R1-MYB transcription factor, designated as JcR1MYB1, was isolated from Jatropha curcas using rapid amplification of cDNA ends. JcR1MYB1 contains a 951 bp open reading frame that encodes 316 amino acids. The deduced JcR1MYB1 protein was predicted to possess the conserved, 56-amino acid-long DNA-binding domain, which consists of a single helix-turn-helix module and usually occurs in R1-MYBs. JcR1MYB1 is a member of the R1-MYB transcription factor subfamily. A subcellular localization study confirmed the nuclear localization of JcR1MYB1. Expression analysis showed that JcR1MYB1 transcripts accumulated in various examined tissues, with high expression levels in the root and low levels in the stem. JcR1MYB1 transcription was up-regulated by polyethylene glycol, NaCl, and cold treatments, as well as by abscisic acid, jasmonic acid, and ethylene treatment. Analysis of transgenic tobacco plants over-expressing JcR1MYB1 indicates an inportant function for this gene in salt stress.

  12. Development of New Modular Genetic Tools for Engineering the Halophilic Archaeon Halobacterium salinarum.

    Rafael Silva-Rocha

    Full Text Available Our ability to genetically manipulate living organisms is usually constrained by the efficiency of the genetic tools available for the system of interest. In this report, we present the design, construction and characterization of a set of four new modular vectors, the pHsal series, for engineering Halobacterium salinarum, a model halophilic archaeon widely used in systems biology studies. The pHsal shuttle vectors are organized in four modules: (i the E. coli's specific part, containing a ColE1 origin of replication and an ampicillin resistance marker, (ii the resistance marker and (iii the replication origin, which are specific to H. salinarum and (iv the cargo, which will carry a sequence of interest cloned in a multiple cloning site, flanked by universal M13 primers. Each module was constructed using only minimal functional elements that were sequence edited to eliminate redundant restriction sites useful for cloning. This optimization process allowed the construction of vectors with reduced sizes compared to currently available platforms and expanded multiple cloning sites. Additionally, the strong constitutive promoter of the fer2 gene was sequence optimized and incorporated into the platform to allow high-level expression of heterologous genes in H. salinarum. The system also includes a new minimal suicide vector for the generation of knockouts and/or the incorporation of chromosomal tags, as well as a vector for promoter probing using a GFP gene as reporter. This new set of optimized vectors should strongly facilitate the engineering of H. salinarum and similar strategies could be implemented for other archaea.

  13. Systems analysis of bioenergetics and growth of the extreme halophile Halobacterium salinarum.

    Orland Gonzalez


    Full Text Available Halobacterium salinarum is a bioenergetically flexible, halophilic microorganism that can generate energy by respiration, photosynthesis, and the fermentation of arginine. In a previous study, using a genome-scale metabolic model, we have shown that the archaeon unexpectedly degrades essential amino acids under aerobic conditions, a behavior that can lead to the termination of growth earlier than necessary. Here, we further integratively investigate energy generation, nutrient utilization, and biomass production using an extended methodology that accounts for dynamically changing transport patterns, including those that arise from interactions among the supplied metabolites. Moreover, we widen the scope of our analysis to include phototrophic conditions to explore the interplay between different bioenergetic modes. Surprisingly, we found that cells also degrade essential amino acids even during phototropy, when energy should already be abundant. We also found that under both conditions considerable amounts of nutrients that were taken up were neither incorporated into the biomass nor used as respiratory substrates, implying the considerable production and accumulation of several metabolites in the medium. Some of these are likely the products of forms of overflow metabolism. In addition, our results also show that arginine fermentation, contrary to what is typically assumed, occurs simultaneously with respiration and photosynthesis and can contribute energy in levels that are comparable to the primary bioenergetic modes, if not more. These findings portray a picture that the organism takes an approach toward growth that favors the here and now, even at the cost of longer-term concerns. We believe that the seemingly "greedy" behavior exhibited actually consists of adaptations by the organism to its natural environments, where nutrients are not only irregularly available but may altogether be absent for extended periods that may span several years

  14. 圈C4的(St(r),St(r+1)Gr,St(r+1)-冠的优美性%On the gracefulness of the(St(r), St(r+1)Gr, St(r+1)- corona of the cycle C4

    康芳茂; 吴跃生


    给出了圈C4的(St(r), St(r+1)Gr, St(r+1)-冠的定义,讨论了圈C4的(St(r), St(r+1)Gr, St(r+1)-冠的优美性,用构造性的方法给出了圈C4的(St(r), St(r+1)Gr, St(r+1)-冠的优美标号。%Gave the definition of the (St (r ), St (r +1), Gr , St (r +1)) - corona of the cycle C4.Discussed the gracefulness of the (St (r ), St (r +1), Gr , St (r +1)) - corona of the cycleC4.The graceful labelings was given by the method of construction.

  15. Approach toward enhancement of halophilic protease production by Halobacterium sp. strain LBU50301 using statistical design response surface methodology

    Julalak Chuprom


    Full Text Available A new potent halophilic protease producer, Halobacterium sp. strain LBU50301 was isolated from salt-fermented fish samples (budu and identified by phenotypic analysis, and 16S rDNA gene sequencing. Thereafter, sequential statistical strategy was used to optimize halophilic protease production from Halobacterium sp. strain LBU50301 by shake-flask fermentation. The classical one-factor-at-a-time (OFAT approach determined gelatin was the best nitrogen source. Based on Plackett–Burman (PB experimental design; gelatin, MgSO4·7H2O, NaCl and pH significantly influenced the halophilic protease production. Central composite design (CCD determined the optimum level of medium components. Subsequently, an 8.78-fold increase in corresponding halophilic protease yield (156.22 U/mL was obtained, compared with that produced in the original medium (17.80 U/mL. Validation experiments proved the adequacy and accuracy of model, and the results showed the predicted value agreed well with the experimental values. An overall 13-fold increase in halophilic protease yield was achieved using a 3 L laboratory fermenter and optimized medium (231.33 U/mL.

  16. miR-1 exacerbates cardiac ischemia-reperfusion injury in mouse models.

    Zhenwei Pan

    Full Text Available Recent studies have revealed the critical role of microRNAs (miRNAs in regulating cardiac injury. Among them, the cardiac enriched microRNA-1(miR-1 has been extensively investigated and proven to be detrimental to cardiac myocytes. However, solid in vivo evidence for the role of miR-1 in cardiac injury is still missing and the potential therapeutic advantages of systemic knockdown of miR-1 expression remained unexplored. In this study, miR-1 transgenic (miR-1 Tg mice and locked nucleic acid modified oligonucleotide against miR-1 (LNA-antimiR-1 were used to explore the effects of miR-1 on cardiac ischemia/reperfusion injury (30 min ischemia followed by 24 h reperfusion. The cardiac miR-1 level was significantly increased in miR-1 Tg mice, and suppressed in LNA-antimiR-1 treated mice. When subjected to ischemia/reperfusion injury, miR-1 overexpression exacerbated cardiac injury, manifested by increased LDH, CK levels, caspase-3 expression, apoptosis and cardiac infarct area. On the contrary, LNA-antimiR-1 treatment significantly attenuated cardiac ischemia/reperfusion injury. The expression of PKCε and HSP60 was significantly repressed by miR-1 and enhanced by miR-1 knockdown, which may be a molecular mechanism for the role miR-1 in cardiac injury. Moreover, luciferase assay confirmed the direct regulation of miR-1 on protein kinase C epsilon (PKCε and heat shock protein 60 (HSP60. In summary, this study demonstrated that miR-1 is a causal factor for cardiac injury and systemic LNA-antimiR-1 therapy is effective in ameliorating the problem.

  17. CSIR wins R1 million gas generator contract


    The Division of Energy Technology, CSIR, has signed a R1 million contract with Slagment (Pty) Ltd for the supply, installation and commissioning during the first half of 1989 of a fluidized-bed hot gas generator. The contract was won on selected tender. A demonstration plant was built during 1984 in Pretoria West and funded by the Development of Mineral and Energy Affairs, to demonstrate to industry the ability of fluidized-bed technology to burn discard coal, thus reducing energy costs. It has been in operation for more than 20 000 hours and has demonstrated that it can successfully operate on coals with up to 70% ash and 8% sulfur content as well as on duff coal and char with a fines content of up to 50% less than 2mm diameter. The importance of offering a solution to the serious problem of stockpiling local inferior coals and the prospect of developing local expertise in the new, emerging technology, gave the research project the required support. The following are discussed: fluidization, fluidized beds and combustion of coal, fluidized bed boilers burning coal, fluidized-bed hot gas generator burning coal; advantages of fluidized-bed combustors, and applications. 2 figs.

  18. New burnup calculation of TRIGA IPR-R1 reactor

    Meireles, Sincler P. de; Campolina, Daniel de A.M.; Santos, Andre A. Campagnole dos; Menezes, Maria A.B.C.; Mesquita, Amir Z., E-mail: [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)


    The IPR-R1 TRIGA Mark I research reactor, located at the Nuclear Technology Development Center - CDTN, Belo Horizonte, Brazil, operates since 1960.The reactor is operating for more than fifty years and has a long history of operation. Determining the current composition of the fuel is very important to calculate various parameters. The reactor burnup calculation has been performed before, however, new techniques, methods, software and increase of the processing capacity of the new computers motivates new investigations to be performed. This work presents the evolution of effective multiplication constant and the results of burnup. This new model has a more detailed geometry with the introduction of the new devices, like the control rods and the samarium discs. This increase of materials in the simulation in burnup calculation was very important for results. For these series of simulations a more recently cross section library, ENDF/B-VII, was used. To perform the calculations two Monte Carlo particle transport code were used: Serpent and MCNPX. The results obtained from two codes are presented and compared with previous studies in the literature. (author)

  19. The photochemical reaction cycle and photoinduced proton transfer of sensory rhodopsin II (Phoborhodopsin) from Halobacterium salinarum.

    Tamogami, Jun; Kikukawa, Takashi; Ikeda, Yoichi; Takemura, Ayaka; Demura, Makoto; Kamo, Naoki


    Sensory rhodopsin II (HsSRII, also called phoborhodopsin) is a negative phototaxis receptor of Halobacterium salinarum, a bacterium that avoids blue-green light. In this study, we expressed the protein in Escherichia coli cells, and reconstituted the purified protein with phosphatidylcholine. The reconstituted HsSRII was stable. We examined the photocycle by flash-photolysis spectroscopy in the time range of milliseconds to seconds, and measured proton uptake/release using a transparent indium-tin oxide electrode. The pKa of the counterion of the Schiff base, Asp(73), was 3.0. Below pH 3, the depleted band was observed on flash illumination, but the positive band in the difference spectra was not found. Above pH 3, the basic photocycle was HsSRII (490) --> M (350) --> O (520) --> Y (490) --> HsSRII, where the numbers in parentheses are the maximum wavelengths. The decay rate of O-intermediate and Y-intermediate were pH-independent, whereas the M-intermediate decay was pH-dependent. For 3 decay was one phase, and the rate decreased with an increase in pH. For 4.5 decay was one phase with pH-independent rates, and azide markedly accelerated the M-decay. These findings suggest the existence of a protonated amino acid residue (X-H) that may serve as a proton relay to reprotonate the Schiff base. Above pH 6.5, the M-decay showed two phases. The fast M-decay was pH-independent and originated from the molecule having a protonated X-H, and the slow M-decay originated from the molecule having a deprotonated X, in which the proton came directly from the outside. The analysis yielded a value of 7.5 for the pKa of X-H. The proton uptake and release occurred during M-decay and O-decay, respectively.

  20. 26 CFR 1.414(r)-1 - Requirements applicable to qualified separate lines of business.


    ... lines of business. 1.414(r)-1 Section 1.414(r)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT... Plans, Etc. § 1.414(r)-1 Requirements applicable to qualified separate lines of business. (a) In general. Section 414(r) prescribes the conditions under which an employer is treated as operating...

  1. The Maize enr System of r1 Haplotype–Specific Aleurone Color Enhancement Factors

    We describe a family of three dominant r1 haplotype-specific enhancers of aleurone color in Zea mays. Stable alleles of the three enhancement of r1 loci (enr1, enr2 and enr3) intensify aleurone color conferred by certain pale and near-colorless r1 haplotypes. In addition, unstable alleles of enr1 ac...

  2. Effects of 60Co gamma-rays, ultraviolet light, and mitomycin C on Halobacterium salinarium and Thiobacillus intermedius.

    Shahmohammadi, H R; Asgarani, E; Terato, H; Ide, H; Yamamoto, O


    Lethal effects of 60Co gamma-rays, UV light, and mitomycin C on two kinds of bacteria, Halobacterium salinarium which grows in highly concentrated salt media and Thiobacillus intermedius which requires reduced sulfur compounds, were studied and compared with those on Escherichia coli B/r. D37 values for H. salinarium, T. intermedius and E. coli B/r were 393, 150, and 92 Gy, respectively, by exposure to 60Co gamma-rays. They were 212, 38, and 10 J/m2, respectively, by exposure to UV light and 2.36, 0.25, and 0.53 microgram/ml/h, respectively, by exposure to mitomycin C. Against these agents, H. salinarium was much more resistant than T. intermedius and E. coli B/r.

  3. Primary structures of three highly acidic ribosomal proteins S6, S12 and S15 from the archaebacterium Halobacterium marismortui.

    Kimura, J; Arndt, E; Kimura, M


    The amino acid sequences of three extremely acidic ribosomal proteins, S6, S12, and S15, from Halobacterium marismortui have been determined. The sequences were obtained by the sequence analysis of peptides derived by enzymatic digestion with trypsin. Stapylococcus aureus protease and chymotrypsin, as well as by cleavage with dilute HCl. The proteins, S6, S12 and S15, consist of 116, 147 and 102 amino acid residues, and have molecular masses of 12,251, 16,440 and 11,747 Da, respectively. Comparison of the amino acid sequences of these proteins with ribosomal protein sequences of other organisms revealed that halobacterial protein S12 has homology with the eukaryotic protein S16A from Saccharomyces cerevisiae, while S15 is significantly related to the Xenopus laevis S19 protein. No homology was found between these halobacterial proteins and any eubacterial ribosomal proteins.

  4. Optimization of hydrogen production by Halobacterium salinarium coupled with E coli using milk plasma as fermentative substrate

    Brijesh Prasad


    Full Text Available Batch experiments were conducted to investigate the fermentative hydrogen production by coupled system of Halobacterium salinarium and E. coli. Increase in the light intensity from 6000 lux to 12000 lux and changing the inoculums level of E coli resulted in 10 fold increase in the rate of hydrogen production using the coupled system. Statistical based design of experiments was applied to optimize the rate of hydrogen production using milk plasma,popularly known as cheese whey, a dairy industry byproduct. An optimal rate of hydrogen production of 56.7 ml/l h was achieved with 14.42 % (by volume of milk plasma and an initial pH of 6.6. The investigations provided information on achieving higher yields with milk plasma as substrate, its optimal concentration, and importance of media pH for producing higher rate of hydrogen.

  5. Studies with the human cohesin establishment factor, ChlR1. Association of ChlR1 with Ctf18-RFC and Fen1.

    Farina, Andrea; Shin, Jae-Ho; Kim, Do-Hyung; Bermudez, Vladimir P; Kelman, Zvi; Seo, Yeon-Soo; Hurwitz, Jerard


    Human ChlR1 (hChlR1), a member of the DEAD/DEAH subfamily of helicases, was shown to interact with components of the cohesin complex and play a role in sister chromatid cohesion. In order to study the biochemical and biological properties of hChlR1, we purified the protein from 293 cells and demonstrated that hChlR1 possesses DNA-dependent ATPase and helicase activities. This helicase translocates on single-stranded DNA in the 5' to 3' direction in the presence of ATP and, to a lesser extent, dATP. Its unwinding activity requires a 5'-singlestranded region for helicase loading, since flush-ended duplex structures do not support unwinding. The helicase activity of hChlR1 is capable of displacing duplex regions up to 100 bp, which can be extended to 500 bp by RPA or the cohesion establishment factor, the Ctf18-RFC (replication factor C) complex. We show that hChlR1 interacts with the hCtf18-RFC complex, human proliferating cell nuclear antigen, and hFen1. The interactions between Fen1 and hChlR1 stimulate the flap endonuclease activity of Fen1. Selective depletion of either hChlR1 or Fen1 by targeted small interfering RNA treatment results in the precocious separation of sister chromatids. These findings are consistent with a role of hChlR1 in the establishment of sister chromatid cohesion and suggest that its action may contribute to lagging strand processing events important in cohesion.

  6. Purple membranes from Halobacterium salinarum as building blocks for nanobiotechnology: The importance of mechanical and thermal properties for matrix and surface applications

    Rhinow, Daniel Christopher


    Bacteriorhodopsin (BR) is a light-driven proton pump and the key protein in halobacterial photosynthesis. In its native host, the archaeon Halobacterium salinarum, BR trimers arrange into a 2-D crystalline lattice, the so-called purple membranes (PMs) which comprise BR and lipids only. Along with the PM assembly BR is astonishingly stable against thermal and chemical stress which makes it an excellent candidate for a variety of ...

  7. LPA法克隆嗜盐菌Halobacterium species xz515古紫质基因%Rapid Cloning of an Archaerhodopsin Gene from Halobacterium species xz515 by LPA

    王奕然; 洪洁; 明明; 丁建东; 李庆国; 黄伟达


    Ligation-mediated PCR amplification (LPA) is a novel approach developed for rapid cloning of genes starting from partial known nucleotide sequence. In this report, the application of LPA in the cloning of an archaerhodopsin (AR4) gene from Halobacterium species xz515 (H.sp.xz515) is presented. Bacterial genomic DNA samples were first digested by a group of selected restriction enzymes, separately. The resulting digests were then ligated to one of the three types of specially designed adapters corresponding to the restriction enzymes used for digestion. The ligated DNA samples were mixed together and used as the template for semi-nested PCR amplification of the unknown regions by using the common sequence of the adapters as one of the primers. The full-length AR4 gene was obtained by simply joining the sequences derived by LPA. LPA is ideally suited for rapid cloning of promoter regions of eukaryotic genes departing from its cDNA sequence, as well as routine gene cloning works starting from EST sequences.%Halobacterium species xz515是从中国西藏分离到的嗜盐菌,所含古紫质(古细菌视紫红质的简称命名为AR4) 具有与其他已知菌视紫红质相反的质子释放和吸收的顺序而引起重视. 连接反应介导的PCR扩增法(LPA)是一种克隆部分序列已知的基因的新型克隆方法. LPA法利用"酶切-连接-PCR"之组合,首先选定一组限制性内切酶,各自单独地酶切细菌总DNA样品,然后酶切产物分别与相应的寡聚核苷酸片段接头连接. 连接液混合起来,作为扩增未知序列DNA的模板. 通过这种方法,克隆到了包括完整AR4基因开放阅读框和0.4 kb上游调控区域的总长度1.3 kb的DNA片段. 实验结果表明LPA法可以代替传统的构建基因组DNA文库的方法,可以快速有效地获得目标基因相关的序列.

  8. Cloning and Expression of Ama r 1, as a Novel Allergen of Amaranthus retroflexus Pollen

    Payam Morakabati


    Full Text Available Sensitisation to Amaranthus retroflexus pollen is very common in tropical and subtropical countries. In this study we aimed to produce a recombinant allergenic Ole e 1-like protein from the pollen of this weed. To predict cross-reactivity of this allergen (Ama r 1 with other members of the Ole e 1-like protein family, the nucleotide sequence homology of the Ama r 1 was investigated. The expression of Ama r 1 in Escherichia coli was performed by using a pET-21b(+ vector. The IgE-binding potential of recombinant Ama r 1 (rAma r 1 was evaluated by immunodetection and inhibition assays using 26 patients’ sera sensitised to A. retroflexus pollen. The coding sequence of the Ama r 1 cDNA indicated an open reading frame of 507 bp encoding for 168 amino acid residues which belonged to the Ole e 1-like protein family. Of the 26 serum samples, 10 (38.46% had significant specific IgE levels for rAma r 1. Immunodetection and inhibition assays revealed that the purified rAma r 1 might be the same as that in the crude extract. Ama r 1, the second allergen from the A. retroflexus pollen, was identified as a member of the family of Ole e 1-like protein.

  9. BubR1 is modified by sumoylation during mitotic progression.

    Yang, Feikun; Hu, Liyan; Chen, Cheng; Yu, Jianxiu; O'Connell, Christopher B; Khodjakov, Alexey; Pagano, Michele; Dai, Wei


    BubR1 functions as a crucial component that monitors proper chromosome congression and mitotic timing during cell division. We investigated molecular regulation of BubR1 and found that BubR1 was modified by an unknown post-translation mechanism during the cell cycle, resulting in a significant mobility shift on denaturing gels. We termed it BubR1-M as the nature of modification was not characterized. Extended (>24 h) treatment of HeLa cells with a microtubule disrupting agent including nocodazole and taxol or release of mitotic shake-off cells into fresh medium induced BubR1-M. BubR1-M was derived from neither phosphorylation nor acetylation. Ectopic expression coupled with pulling down analyses showed that BubR1-M was derived from SUMO modification. Mutation analysis revealed that lysine 250 was a crucial site for sumoylation. Significantly, compared with the wild-type control, ectopic expression of a sumoylation-deficient mutant of BubR1 induced chromosomal missegregation and mitotic delay. Combined, our study identifies a new type of post-translational modification that is essential for BubR1 function during mitosis.

  10. Prenatal cocaine exposure uncouples mGluR1 from Homer1 and Gq Proteins.

    Kalindi Bakshi

    Full Text Available Cocaine exposure during gestation causes protracted neurobehavioral changes consistent with a compromised glutamatergic system. Although cocaine profoundly disrupts glutamatergic neurotransmission and in utero cocaine exposure negatively affects metabotropic glutamate receptor-type 1 (mGluR1 activity, the effect of prenatal cocaine exposure on mGluR1 signaling and the underlying mechanism responsible for the prenatal cocaine effect remain elusive. Using brains of the 21-day-old (P21 prenatal cocaine-exposed rats, we show that prenatal cocaine exposure uncouples mGluR1s from their associated synaptic anchoring protein, Homer1 and signal transducer, Gq/11 proteins leading to markedly reduced mGluR1-mediated phosphoinositide hydrolysis in frontal cortex (FCX and hippocampus. This prenatal cocaine-induced effect is the result of a sustained protein kinase C (PKC-mediated phosphorylation of mGluR1 on the serine residues. In support, phosphatase treatment of prenatal cocaine-exposed tissues restores whereas PKC-mediated phosphorylation of saline-treated synaptic membrane attenuates mGluR1 coupling to both Gq/11 and Homer1. Expression of mGluR1, Homer1 or Gα proteins was not altered by prenatal cocaine exposure. Collectively, these data indicate that prenatal cocaine exposure triggers PKC-mediated hyper-phosphorylation of the mGluR1 leading to uncoupling of mGluR1 from its signaling components. Hence, blockade of excessive PKC activation may alleviate abnormalities in mGluR1 signaling and restores mGluR1-regulated brain functions in prenatal cocaine-exposed brains.

  11. Prenatal cocaine exposure uncouples mGluR1 from Homer1 and Gq Proteins.

    Bakshi, Kalindi; Parihar, Raminder; Goswami, Satindra K; Walsh, Melissa; Friedman, Eitan; Wang, Hoau-Yan


    Cocaine exposure during gestation causes protracted neurobehavioral changes consistent with a compromised glutamatergic system. Although cocaine profoundly disrupts glutamatergic neurotransmission and in utero cocaine exposure negatively affects metabotropic glutamate receptor-type 1 (mGluR1) activity, the effect of prenatal cocaine exposure on mGluR1 signaling and the underlying mechanism responsible for the prenatal cocaine effect remain elusive. Using brains of the 21-day-old (P21) prenatal cocaine-exposed rats, we show that prenatal cocaine exposure uncouples mGluR1s from their associated synaptic anchoring protein, Homer1 and signal transducer, Gq/11 proteins leading to markedly reduced mGluR1-mediated phosphoinositide hydrolysis in frontal cortex (FCX) and hippocampus. This prenatal cocaine-induced effect is the result of a sustained protein kinase C (PKC)-mediated phosphorylation of mGluR1 on the serine residues. In support, phosphatase treatment of prenatal cocaine-exposed tissues restores whereas PKC-mediated phosphorylation of saline-treated synaptic membrane attenuates mGluR1 coupling to both Gq/11 and Homer1. Expression of mGluR1, Homer1 or Gα proteins was not altered by prenatal cocaine exposure. Collectively, these data indicate that prenatal cocaine exposure triggers PKC-mediated hyper-phosphorylation of the mGluR1 leading to uncoupling of mGluR1 from its signaling components. Hence, blockade of excessive PKC activation may alleviate abnormalities in mGluR1 signaling and restores mGluR1-regulated brain functions in prenatal cocaine-exposed brains.

  12. SLC9A3R1 stimulates autophagy via BECN1 stabilization in breast cancer cells.

    Liu, Hong; Ma, Yan; He, Hong-Wei; Wang, Jia-Ping; Jiang, Jian-Dong; Shao, Rong-Guang


    Autophagy, a self-catabolic process, has been found to be involved in abrogating the proliferation and metastasis of breast cancer. SLC9A3R1 (solute carrier family 9, subfamily A [NHE3, cation proton antiporter 3], member 3 regulator 1), a multifunctional scaffold protein, is involved in suppressing breast cancer cells proliferation and the SLC9A3R1-related signaling pathway regulates the activation of autophagy processes. However, the precise regulatory mechanism and signaling pathway of SLC9A3R1 in the regulation of autophagy processes in breast cancer cells remains unknown. Here, we report that the stability of BECN1, the major component of the autophagic core lipid kinase complex, is augmented in SLC9A3R1-overexpressing breast cancer MDA-MB-231 cells, subsequently stimulating autophagy by attenuating the interaction between BECN1 and BCL2. Initially, we found that SLC9A3R1 partially stimulated autophagy through the PTEN-PI3K-AKT1 signaling cascade in MDA-MB-231 cells. SLC9A3R1 then attenuated the interaction between BECN1 and BCL2 to stimulate the autophagic core lipid kinase complex. Further findings revealed that SLC9A3R1 bound to BECN1 and subsequently blocked ubiquitin-dependent BECN1 degradation. And the deletion of the C-terminal domain of SLC9A3R1 resulted in significantly reduced binding to BECN1. Moreover, the lack of C-terminal of SLC9A3R1 neither reduced the ubiquitination of BECN1 nor induced autophagy in breast cancer cells. The decrease in BECN1 degradation induced by SLC9A3R1 resulted in the activity of autophagy stimulation in breast cancer cells. These findings indicate that the SLC9A3R1-BECN1 signaling pathway participates in the activation of autophagy processes in breast cancer cells.

  13. The discovery and optimization of pyrimidinone-containing MCH R1 antagonists.

    Hertzog, Donald L; Al-Barazanji, Kamal A; Bigham, Eric C; Bishop, Michael J; Britt, Christy S; Carlton, David L; Cooper, Joel P; Daniels, Alex J; Garrido, Dulce M; Goetz, Aaron S; Grizzle, Mary K; Guo, Yu C; Handlon, Anthony L; Ignar, Diane M; Morgan, Ronda O; Peat, Andrew J; Tavares, Francis X; Zhou, Huiqiang


    Optimization of a series of constrained melanin-concentrating hormone receptor 1 (MCH R1) antagonists has provided compounds with potent and selective MCH R1 activity. Details of the optimization process are provided and the use of one of the compounds in an animal model of diet-induced obesity is presented.

  14. Engineering assessment and certification of integrity of the 141-R1U1 tank system

    Graser, D.A. (Science Applications International Corp. (USA))


    This Engineering Assessment and Certification of Integrity of retention tank 141-R1U1 is in response to the requirements of 40 CFR 265.191 for an existing tank system that stores hazardous waste and does not have secondary containment. This technical assessment has been reviewed by an independent, qualified, California registered professional engineer, who has certified the tank system to be adequately designed and compatible with the stored waste so that it will not collapse rupture, or fail. This document will be kept on file at the facility. Onground retention tanks 141-R1O1 and 141-R1O2, which are also part of the 141-R1 retention tank system, do not have secondary containment; consequently, certification documentation for these tanks is not included in this assessment. A discussion of the onground tanks, however, is included in this report to provide a complete description of the 141-R1 retention tank system. 8 refs., 7 figs.

  15. Thermal hydraulic analysis of the IPR-R1 TRIGA reactor; Analise termo-hidraulica do reator TRIGA IPR-R1

    Veloso, Marcelo Antonio [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN), Belo Horizonte, MG (Brazil); Fortini, Maria Auxiliadora [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Dept. de Engenharia Nuclear


    The subchannel approach, normally employed for the analysis of power reactor cores that work under forced convection, have been used for the thermal hydraulic evaluation of a TRIGA Mark I reactor, named IPR-R1, at 250 kW power level. This was accomplished by using the PANTERA-1P subchannel code, which has been conveniently adapted to the characteristics of natural convection of TRIGA reactors. The analysis of results indicates that the steady state operation of IPR-R1 at 250 kW do not imply risks to installations, workers and public. (author)

  16. Visual inspections of the neutron absorber control rods of the IEA-R1 reactor; Inspecoes visuais nas barras absorvedoras de neutrons do reator IEA-R1

    Silva, Jose Eduardo R. da; Terremoto, Luis A.A.; Castanheira, Myrthes; Zeituni, Carlos A.; Damy, Margaret de A. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Centro de Engenharia Nuclear]. E-mail:


    The Fuel Engineering Division at IPEN/CNEN-SP developed facilities for visual inspection of the IEA-R1 fuel elements and neutron absorbing control rod assemblies inside the research reactor pool. This work presents the method of visual inspection performed at IEA-R1 research reactor. These inspections were adopted to evaluate and to follow the state of the Ag-In-Cd control assemblies fabricated at CERCA in 1972 that remain in use at the reactor core. In 1998, 2000 and 20001, visual inspections were performed in these control rod assemblies, which the general conditions were evaluated. (author)

  17. Decreasing TfR1 expression reverses anemia and hepcidin suppression in β-thalassemic mice.

    Li, Huihui; Choesang, Tenzin; Bao, Weili; Chen, Huiyong; Feola, Maria; Garcia-Santos, Daniel; Li, Jie; Sun, Shuming; Follenzi, Antonia; Pham, Petra; Liu, Jing; Zhang, Jinghua; Ponka, Prem; An, Xiuli; Mohandas, Narla; Fleming, Robert; Rivella, Stefano; Li, Guiyuan; Ginzburg, Yelena


    Iron availability for erythropoiesis and its dysregulation in β-thalassemia are incompletely understood. We previously demonstrated that exogenous apo-transferrin leads to more effective erythropoiesis, decreasing erythroferrone and de-repressing hepcidin in β-thalassemic mice. Transferrin-bound iron binding to transferrin receptor 1 (TfR1) is essential for cellular iron delivery during erythropoiesis. We hypothesize that apo-transferrin's effect is mediated via decreased TfR1 expression, and evaluate TfR1 expression in β-thalassemic mice in vivo and in vitro with and without added apo-transferrin. Our findings demonstrate that β-thalassemic erythroid precursors overexpress TfR1, an effect which can be reversed by the administration of exogenous apo-transferrin. In vitro experiments demonstrate that apo-transferrin inhibits TfR1 expression independent of erythropoietin- and iron-related signaling, decreases TfR1 partitioning to reticulocytes during enucleation, and enhances enucleation of defective β-thalassemic erythroid precursors. These findings strongly suggest that overexpressed TfR1 may play a regulatory role contributing to iron overload and anemia in β-thalassemic mice. To evaluate further, we crossed TfR1+/- mice--themselves exhibiting iron-restricted erythropoiesis with increased hepcidin--with β-thalassemic mice. Resultant double-heterozygote mice demonstrate long-term improvement in ineffective erythropoiesis, hepcidin de-repression, and increased erythroid enucleation relative to β-thalassemic mice. Our data demonstrates for the first time that TfR1+/- haplo-insufficiency reverses iron overload specifically in β-thalassemic erythroid precursors. Taken together, decreasing TfR1 expression during β-thalassemic erythropoiesis, either via directly induced haplo-insufficiency or exogenous apo-transferrin, decreases ineffective erythropoiesis and provides an endogenous mechanism to upregulate hepcidin, leading to sustained iron

  18. Understanding the Adaptation of Halobacterium Species NRC-1 to Its Extreme Environment through Computational Analysis of Its Genome Sequence

    Kennedy, Sean P.; Ng, Wailap Victor; Salzberg, Steven L.; Hood, Leroy; DasSarma, Shiladitya


    The genome of the halophilic archaeon Halobacterium sp. NRC-1 and predicted proteome have been analyzed by computational methods and reveal characteristics relevant to life in an extreme environment distinguished by hypersalinity and high solar radiation: (1) The proteome is highly acidic, with a median pI of 4.9 and mostly lacking basic proteins. This characteristic correlates with high surface negative charge, determined through homology modeling, as the major adaptive mechanism of halophilic proteins to function in nearly saturating salinity. (2) Codon usage displays the expected GC bias in the wobble position and is consistent with a highly acidic proteome. (3) Distinct genomic domains of NRC-1 with bacterial character are apparent by whole proteome BLAST analysis, including two gene clusters coding for a bacterial-type aerobic respiratory chain. This result indicates that the capacity of halophiles for aerobic respiration may have been acquired through lateral gene transfer. (4) Two regions of the large chromosome were found with relatively lower GC composition and overrepresentation of IS elements, similar to the minichromosomes. These IS-element-rich regions of the genome may serve to exchange DNA between the three replicons and promote genome evolution. (5) GC-skew analysis showed evidence for the existence of two replication origins in the large chromosome. This finding and the occurrence of multiple chromosomes indicate a dynamic genome organization with eukaryotic character. PMID:11591641

  19. Protective roles of bacterioruberin and intracellular KCl in the resistance of Halobacterium salinarium against DNA-damaging agents

    Shahmohammadi, H.R.; Asgarani, E.; Terato, Hiroaki; Saito, Takeshi; Ohyama, Yoshihiko; Gekko, Kunihiko; Yamamoto, Osamu; Ide, Hiroshi [Hiroshima Univ. (Japan). Faculty of Science


    Halobacterium salinarium, a member of the extremely halophilic archaebacteria, contains a C{sub 50}-carotenoid namely bacterioruberin. We have previously reported the high resistance of this organism against the lethal actions of DNA-damaging agents including ionizing radiation and ultraviolet light (UV). In this study, we have examined whether bacterioruberin and the highly concentrated salts in this bacterium play protective roles against the lethal actions of ionizing radiation, UV, hydrogen peroxide, and mitomycin-C (MMC). The colourless mutant of H. salinarium deficient in bacterioruberin was more sensitive than the red-pigmented wild-type to all tested DNA-damaging agents except MMC. Circular dichroism (CD) spectra of H. salinarium chromosomal DNA at various concentrations of KCl (0-3.5 M) were similar to that of B-DNA, indicating that no conformational changes occurred as a result of high salt concentrations. However, DNA strand-breaks induced by ionizing radiation were significantly reduced by the presence of either bacterioruberin or concentrated KCl, presumably due to scavenging of free radicals. These results suggest that bacterioruberin and intracellular KCl of H. salinarium protect this organism against the lethal effects of oxidative DNA-damaging agents. (author)

  20. Ser/Thr/Tyr protein phosphorylation in the archaeon Halobacterium salinarum--a representative of the third domain of life.

    Michalis Aivaliotis

    Full Text Available In the quest for the origin and evolution of protein phosphorylation, the major regulatory post-translational modification in eukaryotes, the members of archaea, the "third domain of life", play a protagonistic role. A plethora of studies have demonstrated that archaeal proteins are subject to post-translational modification by covalent phosphorylation, but little is known concerning the identities of the proteins affected, the impact on their functionality, the physiological roles of archaeal protein phosphorylation/dephosphorylation, and the protein kinases/phosphatases involved. These limited studies led to the initial hypothesis that archaea, similarly to other prokaryotes, use mainly histidine/aspartate phosphorylation, in their two-component systems representing a paradigm of prokaryotic signal transduction, while eukaryotes mostly use Ser/Thr/Tyr phosphorylation for creating highly sophisticated regulatory networks. In antithesis to the above hypothesis, several studies showed that Ser/Thr/Tyr phosphorylation is also common in the bacterial cell, and here we present the first genome-wide phosphoproteomic analysis of the model organism of archaea, Halobacterium salinarum, proving the existence/conservation of Ser/Thr/Tyr phosphorylation in the "third domain" of life, allowing a better understanding of the origin and evolution of the so-called "Nature's premier" mechanism for regulating the functional properties of proteins.

  1. Hydrogen gas production by combined systems of Rhodobacter sphaeroides O.U.001 and Halobacterium salinarum in a photobioreactor

    Zabut, Baker; El-Kahlout, Kamal [Department of Biochemistry, School of Science, IUG, Gaza (PS); Yuecel, Meral [Department of Biology, Middle East Technical University, 06531 Ankara (Turkey); Guenduez, Ufuk; Tuerker, Lemi [Department of Chemistry, Middle East Technical University, 06531 Ankara (Turkey); Eroglu, Inci [Department of Chemical Engineering, Middle East Technical University, 06531 Ankara (Turkey)


    Rhodobacter sphaeroides O.U.001 is a photosynthetic non-sulfur bacterium which produces hydrogen from organic compounds under anaerobic conditions. Halobacterium salinarum is an archaeon and lives under extremely halophilic conditions (4M NaCl). H. salinarum contains a retinal protein bacteriorhodopsin in its purple membrane which acts as a light-driven proton pump. In this study the Rhodobacter sphaeroides O.U.001 culture was combined with different amounts of packed cells of H. salinarum S9 or isolated purple membrane fragments in order to increase the photofermentative hydrogen gas production. The packed cells of H. salinarum have the ability to pump protons upon illumination due to the presence of bacteriorhodopsin. The proton gradient produced may be used for the formation of ATP or protons may be used for H{sub 2} production by R. sphaeroides. Similar to intact cells purple membrane fragments may also form vesicles around certain ions and may act like closed systems. The hydrogen production experiments were carried out using 400ml water-jacketed-glass column stirred photobioreactors. In combined systems 10-200nmol of bacteriorhodopsin was used. Hydrogen gas production was enhanced by four- to sixfold in combined systems of H. salinarum packed cells with R. sphaeroides O.U.001 cell. Stirring both increased the total gas produced and enhanced the rate of hydrogen production. The light energy conversion efficiency was increased from 0.6% to 2.25% in combined systems. (author)

  2. A small basic protein from the brz-brb operon is involved in regulation of bop transcription in Halobacterium salinarum

    Dyall-Smith Mike


    Full Text Available Abstract Background The halophilic archaeon Halobacterium salinarum expresses bacteriorhodopsin, a retinal-protein that allows photosynthetic growth. Transcription of the bop (bacterioopsin gene is controlled by two transcription factors, Bat and Brz that induce bop when cells are grown anaerobically and under light. Results A new gene was identified that is transcribed together with the brz gene that encodes a small basic protein designated as Brb (bacteriorhodopsin-regulating basic protein. The translation activity of the start codon of the brb gene was confirmed by BgaH reporter assays. In vivo site-directed mutagenesis of the brb gene showed that the Brb protein cooperates with Brz in the regulation of bop expression. Using a GFP reporter assay, it was demonstrated that Brb cooperates with both Brz and Bat proteins to activate bop transcription under phototrophic growth conditions. Conclusions The activation of the bop promoter was shown to be dependent not only on two major factors, Bat and Brz, but is also tuned by the small basic protein, Brb.

  3. RyR1 S-nitrosylation underlies environmental heat stroke and sudden death in Y522S RyR1 knockin mice.

    Durham, William J; Aracena-Parks, Paula; Long, Cheng; Rossi, Ann E; Goonasekera, Sanjeewa A; Boncompagni, Simona; Galvan, Daniel L; Gilman, Charles P; Baker, Mariah R; Shirokova, Natalia; Protasi, Feliciano; Dirksen, Robert; Hamilton, Susan L


    Mice with a malignant hyperthermia mutation (Y522S) in the ryanodine receptor (RyR1) display muscle contractures, rhabdomyolysis, and death in response to elevated environmental temperatures. We demonstrate that this mutation in RyR1 causes Ca(2+) leak, which drives increased generation of reactive nitrogen species (RNS). Subsequent S-nitrosylation of the mutant RyR1 increases its temperature sensitivity for activation, producing muscle contractures upon exposure to elevated temperatures. The Y522S mutation in humans is associated with central core disease. Many mitochondria in the muscle of heterozygous Y522S mice are swollen and misshapen. The mutant muscle displays decreased force production and increased mitochondrial lipid peroxidation with aging. Chronic treatment with N-acetylcysteine protects against mitochondrial oxidative damage and the decline in force generation. We propose a feed-forward cyclic mechanism that increases the temperature sensitivity of RyR1 activation and underlies heat stroke and sudden death. The cycle eventually produces a myopathy with damaged mitochondria.

  4. Separating the spindle, checkpoint, and timer functions of BubR1.

    Rahmani, Zohra; Gagou, Mary E; Lefebvre, Christophe; Emre, Doruk; Karess, Roger E


    BubR1 performs several roles during mitosis, affecting the spindle assembly checkpoint (SAC), mitotic timing, and spindle function, but the interdependence of these functions is unclear. We have analyzed in Drosophila melanogaster the mitotic phenotypes of kinase-dead (KD) BubR1 and BubR1 lacking the N-terminal KEN box. bubR1-KD individuals have a robust SAC but abnormal spindles with thin kinetochore fibers, suggesting that the kinase activity modulates microtubule capture and/or dynamics but is relatively dispensable for SAC function. In contrast, bubR1-KEN flies have normal spindles but no SAC. Nevertheless, mitotic timing is normal as long as Mad2 is present. Thus, the SAC, timer, and spindle functions of BubR1 are substantially separable. Timing is shorter in bubR1-KEN mad2 double mutants, yet in these flies, lacking both critical SAC components, chromosomes still segregate accurately, reconfirming that in Drosophila, reliable mitosis does not need the SAC.

  5. On entire f-maximal graphs in the Lorentzian product Gn ×R1

    An, H. V. Q.; Cuong, D. V.; Duyen, N. T. M.; Hieu, D. T.; Nam, T. L.


    In the Lorentzian product Gn ×R1, we give a comparison theorem between the f-volume of an entire f-maximal graph and the f-volume of the hyperbolic Hr+ under the condition that the gradient of the function defining the graph is bounded away from 1. This condition comes from an example of non-planar entire f-maximal graph in Gn ×R1 and is equivalent to the hyperbolic angle function of the graph being bounded. As a consequence, we obtain a Calabi-Bernstein type theorem for f-maximal graphs in Gn ×R1.

  6. Research on metabolic network of extremely halophilic archaea%极端嗜盐古菌代谢网络研究



    Extremely halophilic archaea belonging to the euryarchaeota halobium branch, it is a typical population in archaea, and very important to microbial resources. In this article, we mainly study the structure and function of metabolic network about one extremely halophilic archaea: Halobacterium salinarum Rl. We obtain a list of all metabolic reactions in Halobacterium salinarum Rl from published high-quality metabolic models, and represent it with metabolite graph. We analyze functional modules and hubs of the network, and discussed their biological signification.%极端嗜盐古菌(extremely thermophilic archaea)属于广域古菌界盐杆菌科,是古菌域中的典型生理类群,也是重要的极端微生物资源.文章主要对一种极端嗜盐古菌——盐沼盐杆菌(Halobacterium salinarum R1)——代谢网络的结构与功能进行了分析.对高质量盐沼盐杆菌代谢网络数据进行整理,构建了其中所有的代谢反应列表,并用代谢物图来表示.分析了该网络的功能模块和关键节点,并讨论了它们的生物学功能意义.

  7. Iron-oxo clusters biomineralizing on protein surfaces: Structural analysis of Halobacterium salinarum DpsA in its low- and high-iron states

    Zeth, Kornelius; Offermann, Stefanie; Essen, Lars-Oliver; Oesterhelt, Dieter


    The crystal structure of the Dps-like (Dps, DNA-protecting protein during starvation) ferritin protein DpsA from the halophile Halobacterium salinarum was determined with low endogenous iron content at 1.6-Å resolution. The mechanism of iron uptake and storage was analyzed in this noncanonical ferritin by three high-resolution structures at successively increasing iron contents. In the high-iron state of the DpsA protein, up to 110 iron atoms were localized in the dodecameric protein complex....

  8. New Salmonella serotype: Salmonella enteritidis serotype Grandhaven (30(1):r:1,2).

    McDougal, D L; Treleaven, B E; Renshaw, E C


    A new Salmonella serotype, Salmonella enteritidis serotype Grandhaven (30(1):r:1,2), was isolated from the stool of a 35-year-old man with mild gastroenteritis. He had just returned from Sudan, Africa.

  9. Mammalian ChlR1 has a role in heterochromatin organization

    Inoue, Akira; Hyle, Judith [Department of Tumor Cell Biology, St. Jude Children' s Research Hospital, Memphis, TN (United States); Lechner, Mark S. [Faculty of Health Sciences, Simon Fraser University, Burnaby, BC (Canada); Lahti, Jill M., E-mail: [Department of Tumor Cell Biology, St. Jude Children' s Research Hospital, Memphis, TN (United States); Department of Molecular Sciences, University of Tennessee Health Science Center, Memphis, TN (United States)


    The ChlR1 DNA helicase, encoded by DDX11 gene, which is responsible for Warsaw breakage syndrome (WABS), has a role in sister-chromatid cohesion. In this study, we show that human ChlR1 deficient cells exhibit abnormal heterochromatin organization. While constitutive heterochromatin is discretely localized at perinuclear and perinucleolar regions in control HeLa cells, ChlR1-depleted cells showed dispersed localization of constitutive heterochromatin accompanied by disrupted centromere clustering. Cells isolated from Ddx11{sup -/-} embryos also exhibited diffuse localization of centromeres and heterochromatin foci. Similar abnormalities were found in HeLa cells depleted of combinations of HP1{alpha} and HP1{beta}. Immunofluorescence and chromatin immunoprecipitation showed a decreased level of HP1{alpha} at pericentric regions in ChlR1-depleted cells. Trimethyl-histone H3 at lysine 9 (H3K9-me3) was also modestly decreased at pericentric sequences. The abnormality in pericentric heterochromatin was further supported by decreased DNA methylation within major satellite repeats of Ddx11{sup -/-} embryos. Furthermore, micrococcal nuclease (MNase) assay revealed a decreased chromatin density at the telomeres. These data suggest that in addition to a role in sister-chromatid cohesion, ChlR1 is also involved in the proper formation of heterochromatin, which in turn contributes to global nuclear organization and pleiotropic effects. -- Highlights: {yields} New role for ChlR1 (DDX11), a cohesinopathy gene, in heterochromatin organization. {yields} Loss of ChlR1 altered heterochromatin localization and centromere clustering. {yields} Reduced ChlR1 levels also reduced HP1{alpha} and H3K9-me3 binding to pericentric DNA. {yields} Decreased DNA methylation was found in pericentric repeats of Ddx11{sup -/-} embryos. {yields} These findings will aid in understanding the pathogenesis of Warsaw breakage syndrome.

  10. Unified segmentation based correction of R1 brain maps for RF transmit field inhomogeneities (UNICORT).

    Weiskopf, Nikolaus; Lutti, Antoine; Helms, Gunther; Novak, Marianne; Ashburner, John; Hutton, Chloe


    Quantitative mapping of the longitudinal relaxation rate (R1=1/T1) in the human brain enables the investigation of tissue microstructure and macroscopic morphology which are becoming increasingly important for clinical and neuroimaging applications. R1 maps are now commonly estimated from two fast high-resolution 3D FLASH acquisitions with variable excitation flip angles, because this approach is fast and does not rely on special acquisition techniques. However, these R1 maps need to be corrected for bias due to RF transmit field (B1(+)) inhomogeneities, requiring additional B1(+) mapping which is usually time consuming and difficult to implement. We propose a technique that simultaneously estimates the B1(+) inhomogeneities and R1 values from the uncorrected R1 maps in the human brain without need for B1(+) mapping. It employs a probabilistic framework for unified segmentation based correction of R1 maps for B1(+) inhomogeneities (UNICORT). The framework incorporates a physically informed generative model of smooth B1(+) inhomogeneities and their multiplicative effect on R1 estimates. Extensive cross-validation with the established standard using measured B1(+) maps shows that UNICORT yields accurate B1(+) and R1 maps with a mean deviation from the standard of less than 4.3% and 5%, respectively. The results of different groups of subjects with a wide age range and different levels of atypical brain anatomy further suggest that the method is robust and generalizes well to wider populations. UNICORT is easy to apply, as it is computationally efficient and its basic framework is implemented as part of the tissue segmentation in SPM8.

  11. miR-1 Inhibits Cell Growth, Migration, and Invasion by Targeting VEGFA in Osteosarcoma Cells

    Junjie Niu


    Full Text Available microRNAs (miRNAs are small noncoding RNAs and have been shown to play a crucial role in the osteosarcoma (OS tumorigenesis and progression. VEGFA is a key regulator of angiogenesis and plays an important role in regulation of tumor metastasis. The objective of this study was to determine whether VEGFA was involved in miR-1-mediated suppression of proliferation, migration, and invasion of OS cells. The expression levels of miR-1 were significantly lower in OS tumor tissues than those in adjacent normal tissues and in SAOS-2 and U2OS cell lines compared to a normal osteoblast (NHOst cell line. VEGFA was upregulated in OS tumor tissues and SAOS-2 and U2OS cell lines. The results of CCK-8 assay and transwell assay showed that miR-1 acted as a tumor suppressor by inhibiting cell proliferation, migration, and invasion in U2OS cells. Dual luciferase reporter assay demonstrated that VEGFA was a direct and functional target gene of miR-1. miR-1 directly inhibits the protein expression of VEGFA via its 3′-UTR. Knockdown of VEGFA by siRNA inhibited proliferation, migration, and invasion of U2OS cells. Our study suggested the potential inhibitory function of miR-1 in OS cell proliferation, migration, and invasion via inhibiting VEGFA.

  12. Regulation of the cardioprotective adiponectin and its receptor AdipoR1 by salt.

    Arnold, Nicholas; Mahmood, Abuzar; Ramdas, Maya; Ehlinger, Paul P; Pulakat, Lakshmi


    Both circulating adiponectin (APN) and cardiac APN exert cardioprotective effects and improve insulin sensitivity and mitochondrial function. Low circulating APN serves as a biomarker for cardiovascular risk. Ablation of adiponectin receptor 1 (AdipoR1) causes myocardial mitochondrial dysfunction. Although high salt intake is a contributor to cardiovascular disease, how it modulates the expression of APN or AdipoR1 in cardiomyocytes is not known. We report that APN mRNA expression was attenuated in a dose-dependent manner in mouse cardiomyocyte cell line HL-1 exposed to salt concentrations ranging from 0.75% to 1.5% for 12 h. High-salt exposure (0.88% and 1.25% for 12 h) also suppressed APN and AdipoR1 protein expression significantly in rat cardiac muscle H9c2 cells. Co-immunostaining for AdipoR1 and mitochondrial complex 1 indicated that AdipoR1 may be co-localized with mitochondria. These data show for the first time that high salt is an important suppressor of cardiovascular protective APN and AdipoR1.

  13. IEA-R1 reactor spent fuel element surveillance; Acompanhamento da irradiacao dos elementos combustiveis do reator IEA-R1

    Damy, Margaret de Almeida; Terremoto, Luis Antonio Albiac; Silva, Jose Eduardo Rosa da; Silva, Antonio Teixeira e; Teodoro, Celso A.; Lucki, Georgi; Castanheira, Myrthes [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail:


    The irradiation surveillance is an important part of a qualification program of the U{sub 3}O{sub 8}-Al and U{sub 3}Si{sub 2}-Al dispersion nuclear fuels manufactured in IPEN/CNEN-SP. This work presents the surveillance results regarding the fuel and control elements irradiated in the IEA-R1 research reactor during the period from June/1999 until December/2003, which embraced register of visual inspections, irradiation conditions, burn-up calculations, thermal hydraulic parameters and failure occurrences. Also providing information that helps the safe operation of the IEA-R1 research reactor, the irradiation surveillance is a collaboration work involving researchers of the Centro de Engenharia Nuclear (CEN) and the operators' staff of the Centro do Reator de Pesquisas (CRPq), both from IPEN/CNEN-SP. (author)

  14. Aspects of the Iea-R1 research reactor seismic evaluation; Aspectos da avaliacao sismica do reator de pesquisa IEA-R1

    Mattar Neto, Miguel [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)


    Codes and standards for the seismic evaluation of the research reactor IEA-R1 are presented. An approach to define the design basis earthquake based on the local seismic map and on simplified analysis methods is proposed. The site seismic evaluation indicates that the design earthquake intensity is IV MM. Therefore, according to the used codes and standards, no buildings, systems, and components seismic analysis are required. (author)


    Sych I.V.


    Full Text Available Introduction: The analysis of modern literature, including overseas one, showed that a lot of the scientific researches is devoted to finding and creating biologically active compounds on base 1,3,4-thiadiazole. Derivatives of 1,3,4-thiadiazole are the large group of heterocyclic compounds with high rates of antimicrobial, antituberculosis, antidiabetic, antineoplastic and anticonvulsant activity. Material and methods: The purpose of this study was the expansion of sulfone derivatives substituted nitrogen-containing heterocyclic systems through the synthesis of 2-R1-N (5-R-1,3,4-thiadiazol-2-ilbenzolsulfonamides and prediction their pharmacological activity for future planning pharmacological screening. Synthesis of semi-products 2-amino-5-R-1,3,4-thiadiazoles was carried out by cyclization thiosemicarbazide and substituted derivatives of carboxylic acids in the presence of concentrated sulfuric acid. The synthesis of target compounds 2-R1-N(5-R-1,3,4-thiadiazol-2-ylbenzolsulfon-amides was carried out by N-acylation of 2-amino-5R-1,3,4-thiadiazole substituted benzolsul-fochlorides in the presence of anhydrous pyridine. The reaction proceeds by the classic SN2-mechanism. The resulting compounds are white crystalline substances, soluble in alcohol, chloroform and acetone, difficult to dissolve in water. Yields of obtained compounds was satisfactory (76-84%. The purity of the obtained compounds was determined by TLC. The structure of the obtained compounds was proved by elemental analysis, IR methods and 1H NMR spectroscopy. NMR 1H spectra were recorded at Bruker WM spectrometer (200 MHz; solvent DMSO-d6; chemical shifts were in ppm, internal standard (TMS (tetramethylsilane was used. The prognosis of biological activity for obtained compounds were carried out using the program PASS (Prediction of Activity Spectra for Substances in order to plan the further pharmacological screening. The program PASS predicts more than 500 kinds of biological

  16. Alterations in CD200-CD200R1 System during EAE Already Manifest at Presymptomatic Stages

    Tony Valente


    Full Text Available In the brain of patients with multiple sclerosis, activated microglia/macrophages appear in active lesions and in normal appearing white matter. However, whether they play a beneficial or a detrimental role in the development of the pathology remains a controversial issue. The production of pro-inflammatory molecules by chronically activated microglial cells is suggested to contribute to the progression of neurodegenerative processes in neurological disease. In the healthy brain, neurons control glial activation through several inhibitory mechanisms, such as the CD200-CD200R1 interaction. Therefore, we studied whether alterations in the CD200-CD200R1 system might underlie the neuroinflammation in an experimental autoimmune encephalomyelitis (EAE model of multiple sclerosis. We determined the time course of CD200 and CD200R1 expression in the brain and spinal cord of an EAE mouse model from presymptomatic to late symptomatic stages. We also assessed the correlation with associated glial activation, inflammatory response and EAE severity. Alterations in CD200 and CD200R1 expression were mainly observed in spinal cord regions in the EAE model, mostly a decrease in CD200 and an increase in CD200R1 expression. A decrease in the expression of the mRNA encoding a full CD200 protein was detected before the onset of clinical signs, and remained thereafter. A decrease in CD200 protein expression was observed from the onset of clinical signs. By contrast, CD200R1 expression increased at EAE onset, when a glial reaction associated with the production of pro- and anti-inflammatory markers occurred, and continued to be elevated during the pathology. Moreover, the magnitude of the alterations correlated with severity of the EAE mainly in spinal cord. These results suggest that neuronal-microglial communication through CD200-CD200R1 interaction is compromised in EAE. The early decreases in CD200 expression in EAE suggest that this downregulation might also

  17. Characteristics of dibenzothiophene desulfurization by Rhodococcus erythropolis R1 and its Dsz-negative mutant

    Zahra Etemadifar


    Full Text Available Introduction: Biodesulfurization is used as a selective method for lowering the sulfur content of petroleum products. Materials and methods: A sulfur-oxidation bacterial strain named Rhodococcus erythropolis R1 (NCBI GenBank Accession No. GU570564 was used in this study for desulfurization of dibenzothiophene (DBT. Results: The induced culture of strain R1 was able to produce 2-hydroxybiphenyl (2- HBP from DBT followed 4S pathway without further degrading carbon backbone. This process confirmed by gas chromatography (GC analysis. The specific activity of DBT desulfurization by R1 was 45 µM (g dry wt-1 h-1. The addition of Tween 80 as surfactant and glycerol as carbon source determines a 100% rate of DBT-desulfurization during 3 days. The heavy plasmid detected in R1 strain carries dsz genes responsible for biodesulfurization of DBT that was shown by PCR reaction. The mutant strains which had lost this plasmid also had lost desulfurization phenotype. Both mutant and wild strain were sensitive to high concentration of 2-HBP and some antibiotics. Discussion and conclusion: Strain R1 desulfurize DBT through the sulfur-specific degradation pathway or 4S pathway with the selective cleavage of carbon-sulfur (C-S bonds without reducing the energy content. Addition of surfactant enhanced the desulfurization of DBT by increasing its bioavailability and also could improve the growth and desulfurization rate. The location of desulfurization genes was on a heavy plasmid in strain R1. Based on the results of this study, R. erythropolis R1 could serve as a model system for efficient biodesulfurization of petroleum oil without reducing the energy value.

  18. C3a Enhances the Formation of Intestinal Organoids through C3aR1

    Naoya Matsumoto


    Full Text Available C3a is important in the regulation of the immune response as well as in the development of organ inflammation and injury. Furthermore, C3a contributes to liver regeneration but its role in intestinal stem cell function has not been studied. We hypothesized that C3a is important for intestinal repair and regeneration. Intestinal organoid formation, a measure of stem cell capacity, was significantly limited in C3-deficient and C3a receptor (C3aR 1-deficient mice while C3a promoted the growth of organoids from normal mice by supporting Wnt-signaling but not from C3aR1-deficient mice. Similarly, the presence of C3a in media enhanced the expression of the intestinal stem cell marker leucine-rich repeat G-protein-coupled receptor 5 (Lgr5 and of the cell proliferation marker Ki67 in organoids formed from C3-deficient but not from C3aR1-deficient mice. Using Lgr5.egfp mice we showed significant expression of C3 in Lgr5+ intestinal stem cells whereas C3aR1 was expressed on the surface of various intestinal cells. C3 and C3aR1 expression was induced in intestinal crypts in response to ischemia/reperfusion injury. Finally, C3aR1-deficient mice displayed ischemia/reperfusion injury comparable to control mice. These data suggest that C3a through interaction with C3aR1 enhances stem cell expansion and organoid formation and as such may have a role in intestinal regeneration.

  19. Agonistic CD200R1 DNA Aptamers Are Potent Immunosuppressants That Prolong Allogeneic Skin Graft Survival

    Aaron Prodeus


    Full Text Available CD200R1 expressed on the surface of myeloid and lymphoid cells delivers immune inhibitory signals to modulate inflammation when engaged with its ligand CD200. Signalling through CD200/CD200R1 has been implicated in a number of immune-related diseases including allergy, infection, cancer and transplantation, as well as several autoimmune disorders including arthritis, systemic lupus erythematosus, and multiple sclerosis. We report the development and characterization of DNA aptamers, which bind to murine CD200R1 and act as potent signalling molecules in the absence of exogenous CD200. These agonistic aptamers suppress cytotoxic T-lymphocyte induction in 5-day allogeneic mixed leukocyte culture and induce rapid phosphorylation of the CD200R1 cytoplasmic tail thereby initiating immune inhibitory signalling. PEGylated conjugates of these aptamers show significant in vivo immunosuppression and enhance survival of allogeneic skin grafts as effectively as soluble CD200Fc. As DNA aptamers exhibit inherent advantages over conventional protein-based therapeutics including low immunogenicity, ease of synthesis, low cost, and long shelf life, such CD200R1 agonistic aptamers may emerge as useful and safe nonsteroidal anti-inflammatory therapeutic agents.

  20. Protein-Nanocrystal Conjugates Support a Single Filament Polymerization Model in R1 Plasmid Segregation

    Choi, Charina L.; Claridge, Shelley A.; Garner, Ethan C.; Alivisatos, A. Paul; Mullins, R. Dyche


    To ensure inheritance by daughter cells, many low-copy number bacterial plasmids, including the R1 drug-resistance plasmid, encode their own DNA segregation systems. The par operon of plasmid R1 directs construction of a simple spindle structure that converts free energy of polymerization of an actin-like protein, ParM, into work required to move sister plasmids to opposite poles of rod-shaped cells. The structures of individual components have been solved, but little is known about the ultrastructure of the R1 spindle. To determine the number of ParM filaments in a minimal R1 spindle, we used DNA-gold nanocrystal conjugates as mimics of the R1 plasmid. Wefound that each end of a single polar ParM filament binds to a single ParR/parC-gold complex, consistent with the idea that ParM filaments bind in the hollow core of the ParR/parC ring complex. Our results further suggest that multifilament spindles observed in vivo are associated with clusters of plasmidssegregating as a unit.

  1. The AMPA receptor subunit GluR1 regulates dendritic architecture of motor neurons

    Inglis, Fiona M.; Crockett, Richard; Korada, Sailaja; Abraham, Wickliffe C.; Hollmann, Michael; Kalb, Robert G.


    The morphology of the mature motor neuron dendritic arbor is determined by activity-dependent processes occurring during a critical period in early postnatal life. The abundance of the AMPA receptor subunit GluR1 in motor neurons is very high during this period and subsequently falls to a negligible level. To test the role of GluR1 in dendrite morphogenesis, we reintroduced GluR1 into rat motor neurons at the end of the critical period and quantitatively studied the effects on dendrite architecture. Two versions of GluR1 were studied that differed by the amino acid in the "Q/R" editing site. The amino acid occupying this site determines single-channel conductance, ionic permeability, and other essential electrophysiologic properties of the resulting receptor channels. We found large-scale remodeling of dendritic architectures in a manner depending on the amino acid occupying the Q/R editing site. Alterations in the distribution of dendritic arbor were not prevented by blocking NMDA receptors. These observations suggest that the expression of GluR1 in motor neurons modulates a component of the molecular substrate of activity-dependent dendrite morphogenesis. The control of these events relies on subunit-specific properties of AMPA receptors.

  2. Structure of IL-22 Bound to Its High-Affinity IL-22R1 Chain

    Jones, B.C.; Logsdon, N.J.; Walter, M.R. (UAB)


    IL-22 is an IL-10 family cytokine that initiates innate immune responses against bacterial pathogens and contributes to immune disease. IL-22 biological activity is initiated by binding to a cell-surface complex composed of IL-22R1 and IL-10R2 receptor chains and further regulated by interactions with a soluble binding protein, IL-22BP, which shares sequence similarity with an extracellular region of IL-22R1 (sIL-22R1). IL-22R1 also pairs with the IL-20R2 chain to induce IL-20 and IL-24 signaling. To define the molecular basis of these diverse interactions, we have determined the structure of the IL-22/sIL-22R1 complex. The structure, combined with homology modeling and surface plasmon resonance studies, defines the molecular basis for the distinct affinities and specificities of IL-22 and IL-10 receptor chains that regulate cellular targeting and signal transduction to elicit effective immune responses.

  3. GHS-R1a constitutive activity and its physiological relevance

    Yves Louis MEAR


    Full Text Available Abundant evidences have shown that ghrelin, by its binding to GHS-R1a, plays an important role for fundamental physiological functions. Increasing attention is given to the GHS-R1a unusually high constitutive activity and its contribution to downstream signalling and physiological processes. Here, we review recent lines of evidences showing that the interaction between ligand-binding pocket TM domains and the ECL2 could be partially responsible for this high constitutive activity. Interestingly, GHSR-1a constitutive activity activates in turn the downstream PLC, PKC and CRE signalling pathways and this activation is reversed by the inverse agonist [D-Arg1, D-Phe5, D-Trp7,9, Leu11]-substance P (MSP. Noteworthy, GHSR-1a exhibits a C-terminal-dependent constitutive internalization. Non-sense GHS-R1a mutation (Ala204Glu, first discovered in Moroccan patients, supports the role of GHSR-1a constitutive activity in physiological impairments. Ala204Glu-point mutation, altering exclusively the GHSR-1a constitutive activity, was associated with familial short stature syndrome. Altogether, these findings suggest that GHS-R1a constitutive activity could contribute to GH secretion or body weight regulation. Consequently, future research on basic and clinical applications of GHS-R1a inverse agonists will be challenging and potentially rewarding.

  4. Primary identification, biochemical characterization, and immunologic properties of the allergenic pollen cyclophilin cat R 1.

    Ghosh, Debajyoti; Mueller, Geoffrey A; Schramm, Gabriele; Edwards, Lori L; Petersen, Arnd; London, Robert E; Haas, Helmut; Gupta Bhattacharya, Swati


    Cyclophilin (Cyp) allergens are considered pan-allergens due to frequently reported cross-reactivity. In addition to well studied fungal Cyps, a number of plant Cyps were identified as allergens (e.g. Bet v 7 from birch pollen, Cat r 1 from periwinkle pollen). However, there are conflicting data regarding their antigenic/allergenic cross-reactivity, with no plant Cyp allergen structures available for comparison. Because amino acid residues are fairly conserved between plant and fungal Cyps, it is particularly interesting to check whether they can cross-react. Cat r 1 was identified by immunoblotting using allergic patients' sera followed by N-terminal sequencing. Cat r 1 (∼ 91% sequence identity to Bet v 7) was cloned from a cDNA library and expressed in Escherichia coli. Recombinant Cat r 1 was utilized to confirm peptidyl-prolyl cis-trans-isomerase (PPIase) activity by a PPIase assay and the allergenic property by an IgE-specific immunoblotting and rat basophil leukemia cell (RBL-SX38) mediator release assay. Inhibition-ELISA showed cross-reactive binding of serum IgE from Cat r 1-allergic individuals to fungal allergenic Cyps Asp f 11 and Mala s 6. The molecular structure of Cat r 1 was determined by NMR spectroscopy. The antigenic surface was examined in relation to its plant, animal, and fungal homologues. The structure revealed a typical cyclophilin fold consisting of a compact β-barrel made up of seven anti-parallel β-strands along with two surrounding α-helices. This is the first structure of an allergenic plant Cyp revealing high conservation of the antigenic surface particularly near the PPIase active site, which supports the pronounced cross-reactivity among Cyps from various sources.

  5. Fine mapping of the red plant gene R1 in upland cotton(Gossypium hirsutum)

    ZHAO Liang; CAI CaiPing; ZHANG TianZhen; GUO WangZhen


    Sub 16 is a substitution line with G. hirsutum cv. TM-1 genetic background except that the 16th chro-mosome (Chr. 16) is replaced by the corresponding homozygous chromosome of G. barbadense cv. 3-79, and T586 is a G. hirsutum multiple gene marker line with 8 dominant mutation genes. The R1 gene for anthocyanin pigmentation was tagged in Chr. 16 in T586. The objective of this research was to screen SSR markers tightly linked with R1 by using the F2 segregating population containing 1259 plants derived from the cross of Sub 16 and T586 and the backbone genetic linkage map from G. hir-sutumxG, barbadense BC1 newly updated by our laboratory. Genetic analysis suggested that the se-gregation ratio of red plants in the F2 population fit Mendelian 1:2:1 inheritance, confirming that the red plant trait was controlled by an incomplete dominance gene. Preliminary mapping of R1 was conducted using 237 randomLy selected F2 individuals and JoinMap v3.0 software. Then, a fine map of R1 was constructed using the F2 segregating population containing 1259 plants, and R1 was located between NAU4956 and NAU6752, with only 0.49 cM to the nearest maker loci (NAU6752). These results pro-vided a foundation for map-based cloning of R1 and further development of cotton cultivars with red fibers by transgenic technology.

  6. 2-Heteroaryl Benzimidazole Derivatives as Melanin Concentrating Hormone Receptor 1 (MCH-R1) Antagonists

    Lim, Chae Jo; Kim, Jeong Young; Lee, Byung Ho; Oh, Kwangseok; Yi, Kyu Yang [Korea Research Institute of Chemical Technology, Daejeon (Korea, Republic of)


    A novel series of 2-heteroaryl substituted benzimidazole derivatives, containing the piperidinylphenyl acetamide group at the 1-position, were synthesized and evaluated as MCH-R1 antagonists. Extensive SAR investigation probing the effects of C-2 heteroaryl group led to the identification of 2-[2-(pyridin-3-yl)ethyl] analog 3o, which exhibits highly potent MCH-R1 binding activity with an IC{sub 50} value of 1 nM. This substance 3o also has low hERG binding activity, good metabolic stability, and favorable pharmacokinetic properties.

  7. 26 CFR 31.3402(r)-1 - Withholding on distributions of Indian gaming profits to tribal members.


    ... profits to tribal members. 31.3402(r)-1 Section 31.3402(r)-1 Internal Revenue INTERNAL REVENUE SERVICE... TAXES AND COLLECTION OF INCOME TAX AT SOURCE Collection of Income Tax at Source § 31.3402(r)-1 Withholding on distributions of Indian gaming profits to tribal members. (a) (1) General rule. Section...

  8. Direct binding between BubR1 and B56-PP2A phosphatase complexes regulate mitotic progression

    Kruse, Thomas; Zhang, Gang; Larsen, Marie Sofie Yoo;


    and mutation of these residues prevents the establishment of a proper metaphase plate and delays cells in mitosis. Furthermore, we show that phosphorylation of S670 and S676 stimulates the binding of B56 to BubR1 and that BubR1 targets a pool of B56 to kinetochores. Our data suggests that BubR1 counteracts...

  9. Amino acid residues involved in the catalytic mechanism of NAD-dependent glutamate dehydrogenase from Halobacterium salinarum.

    Pérez-Pomares, F; Ferrer, J; Camacho, M; Pire, C; LLorca, F; Bonete, M J


    The pH dependence of kinetic parameters for a competitive inhibitor (glutarate) was determined in order to obtain information on the chemical mechanism for NAD-dependent glutamate dehydrogenase from Halobacterium salinarum. The maximum velocity is pH dependent, decreasing at low pHs giving a pK value of 7.19+/-0.13, while the V/K for l-glutamate at 30 degrees C decreases at low and high pHs, yielding pK values of 7.9+/-0.2 and 9.8+/-0.2, respectively. The glutarate pKis profile decreases at high pHs, yielding a pK of 9. 59+/-0.09 at 30 degrees C. The values of ionization heat calculated from the change in pK with temperature are: 1.19 x 10(4), 5.7 x 10(3), 7 x 10(3), 6.6 x 10(3) cal mol-1, for the residues involved. All these data suggest that the groups required for catalysis and/or binding are lysine, histidine and tyrosine. The enzyme shows a time-dependent loss in glutamate oxidation activity when incubated with diethyl pyrocarbonate (DEPC). Inactivation follows pseudo-first-order kinetics with a second-order rate constant of 53 M-1min-1. The pKa of the titratable group was pK1=6.6+/-0.6. Inactivation with ethyl acetimidate also shows pseudo-first-order kinetics as well as inactivation with TNM yielding second-order constants of 1.2 M-1min-1 and 2.8 M-1min-1, and pKas of 8.36 and 9.0, respectively. The proposed mechanism involves hydrogen binding of each of the two carboxylic groups to tyrosyl residues; histidine interacts with one of the N-hydrogens of the l-glutamate amino group. We also corroborate the presence of a conservative lysine that has a remarkable ability to coordinate a water molecule that would act as general base.

  10. Data of evolutionary structure change: 1DS8R-1AIJS [Confc[Archive

    Full Text Available 1DS8R-1AIJS 1DS8 1AIJ R S ------------ALLSFERKYRVPGGTL-------VGGN...line> LEU CA 264 ALA CA 260 LYS CA 280 1AIJ... S 1AIJS...el> 1 1AIJ S 1AIJ...8992214202881 7.358476161956787 2 1AIJ

  11. Data of evolutionary structure change: 1DS8R-1AIJM [Confc[Archive

    Full Text Available 1DS8R-1AIJM 1DS8 1AIJ R M --------------ALLSFERKYRVPGGTL-------VG...244 SER CA 310 ALA CA 262 VAL CA 196 1AIJ... M 1AIJM LEU CA 264 ALA CA 260 LYS CA 280 1AIJ... M 1AIJM YGLSFAAPLKE

  12. Homotopy classification of maps between r-1 connected 2r dimensional manifolds


    In this paper,we study the homotopy classification of continuous maps between two r-1 connected 2r dimensional topological manifolds M,N.If we assume some knowledge on the homotopy groups of spheres,then the complete classification can be obtained from the homotopy invariants of M,N.We design an algorithm and compose a program to give explicit computations.

  13. Prion pathogenesis is unaltered following down-regulation of SIGN-R1.

    Bradford, Barry M; Brown, Karen L; Mabbott, Neil A


    Prion diseases are infectious neurodegenerative disorders characterised by accumulations of abnormal prion glycoprotein in affected tissues. Following peripheral exposure, many prion strains replicate upon follicular dendritic cells (FDC) in lymphoid tissues before infecting the brain. An intact splenic marginal zone is important for the efficient delivery of prions to FDC. The marginal zone contains a ring of specific intercellular adhesion molecule-3-grabbing non-integrin related 1 (SIGN-R1)-expressing macrophages. This lectin binds dextran and capsular pneumococcal polysaccharides, and also enhances the clearance of apoptotic cells via interactions with complement components. Since prions are acquired as complement-opsonized complexes we determined the role of SIGN-R1 in disease pathogenesis. We show that transient down-regulation of SIGN-R1 prior to intravenous prion exposure had no effect on the early accumulation of prions upon splenic FDC or their subsequent spread to the brain. Thus, SIGN-R1 expression by marginal zone macrophages is not rate-limiting for peripheral prion disease pathogenesis.

  14. Association study of human VN1R1 pheromone receptor gene alleles and gender.

    Mitropoulos, Constantinos; Papachatzopoulou, Adamantia; Menounos, Panagiotis G; Kolonelou, Christina; Pappa, Magda; Bertolis, George; Gerou, Spiros; Patrinos, George P


    Pheromones are water-soluble chemicals that elicit neuroendocrine and physiological changes, while they also provide information about gender within individuals of the same species. VN1R1 is the only functional pheromone receptor in humans. We have undertaken a large mutation screening approach in 425 adult individuals from the Hellenic population to investigate whether the allelic differences, namely alleles 1a and 1b present in the human VN1R1 gene, are gender specific. Here we show that both VN1R1 1a and 1b alleles are found in chromosomes of both male and female subjects at frequency of 26.35% and 73.65%, respectively. Given the fact that those allelic differences potentially cause minor changes in the protein conformation and its transmembrane domains, as simulated by the TMHMM software, our data suggest that the allelic differences in the human VN1R1 gene are unlikely to be associated with gender and hence to contribute to distinct gender-specific behavior.

  15. Homotopy classification of maps between r-1 connected 2r dimensional manifolds

    Xu-an ZHAO; Hong-zhu GAO; Xiao-le SU


    In this paper, we study the homotopy classification of continuous maps between two r - 1 connected 2r dimensional topological manifolds M, N. If we assume some knowledge on the homotopy groups of spheres, then the complete classification can be obtained from the homotopy invariants of M, N. We design an algorithm and compose a program to give explicit computations.

  16. Determination of taste receptor type 1 member 1 (TAS1R1) gene ...



    Oct 10, 2011 ... DOI: 10.5897/AJB11.1379 ... (TAS1R1) gene polymorphism and association with ... 1Institute of Cellular and Molecular Biology, College of Life Science, Xuzhou Normal University, Xuzhou, ... In this study, the coding and flanking .... Note: Exons 1, 2, 3, 4, 5 and 6 are located at base pairs 1~191, 1746~2052, ...

  17. Experience of IEA-R1 research reactor spent fuel transportation back to United States

    Frajndlich, Roberto [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Div. de Operacao do Reator IEAR-R1m]. E-mail:; Perrotta, Jose A. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Engenharia do Nucleo]. E-mail:; Maiorino, Jose Rubens [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Diretoria de Reatores]. E-mail:; Soares, Adalberto Jose [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Dept. de Reatores]. E-mail:


    IPEN/CNEN-SP is sending the IEA-R1 Research Reactor spent fuels from USA origin back to this country. This paper describes the experience in organizing the negotiations, documents and activities to perform the transport. Subjects as cask licensing, transport licensing and fuel failure criteria for transportation are presented. (author)

  18. Highest average burnups achieved by MTR fuel elements of the IEA-R1 research reactor

    Damy, Margaret A.; Terremoto, Luis A.A.; Silva, Jose E.R.; Silva, Antonio Teixeira e; Castanheira, Myrthes; Teodoro, Celso A. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Centro de Engenharia Nuclear (CEN)]. E-mail:


    Different nuclear fuels were employed in the manufacture of plate type at IPEN , usually designated as Material Testing Reactor (MTR) fuel elements. These fuel elements were used at the IEA-R1 research reactor. This work describes the main characteristics of these nuclear fuels, emphasizing the highest average burn up achieved by these fuel elements. (author)

  19. Design of a new wet storage rack for spent fuels from IEA-R1 reactor

    Rodrigues, Antonio C.I.; Madi Filho, Tufic; Siqueira, Paulo T.D.; Ricci Filho, Walter, E-mail:, E-mail:, E-mail:, E-mail: [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    The IEA-R1 research reactor operates in a regimen of 64h weekly, at the power of 4.5 MW. In these conditions, the racks of the spent fuel elements have less than half of its initial capacity. Thus, maintaining these operating conditions, the storage will have capacity for about six years. Since the estimated useful life of the IEA-R1 is about another 20 years, it will be necessary to increase the storage capacity of spent fuel. Dr. Henrik Grahn, expert of the International Atomic Energy Agency on wet storage, visiting the IEA-R1 Reactor (September/2012) made some recommendations: among them, the design and installation of racks made with borated stainless steel and internally coated with an aluminum film, so that corrosion of the fuel elements would not occur. After an extensive literature review of material options given for this type of application we got to Boral® manufactured by 3M due to numerous advantages. This paper presents studies on the analysis of criticality using the computer code MCNP 5, demonstrating the possibility of doubling the storage capacity of current racks to attend the demand of the IEA-R1 reactor while attending the safety requirements the International Atomic Energy Agency. (author)

  20. The Structure and Dynamics of BmR1 Protein from Brugia malayi: In Silico Approaches

    Bee Yin Khor


    Full Text Available Brugia malayi is a filarial nematode, which causes lymphatic filariasis in humans. In 1995, the disease has been identified by the World Health Organization (WHO as one of the second leading causes of permanent and long-term disability and thus it is targeted for elimination by year 2020. Therefore, accurate filariasis diagnosis is important for management and elimination programs. A recombinant antigen (BmR1 from the Bm17DIII gene product was used for antibody-based filariasis diagnosis in “Brugia Rapid”. However, the structure and dynamics of BmR1 protein is yet to be elucidated. Here we study the three dimensional structure and dynamics of BmR1 protein using comparative modeling, threading and ab initio protein structure prediction. The best predicted structure obtained via an ab initio method (Rosetta was further refined and minimized. A total of 5 ns molecular dynamics simulation were performed to investigate the packing of the protein. Here we also identified three epitopes as potential antibody binding sites from the molecular dynamics average structure. The structure and epitopes obtained from this study can be used to design a binder specific against BmR1, thus aiding future development of antigen-based filariasis diagnostics to complement the current diagnostics.

  1. Asymptotic behaviour of H~p (R~n×R_+) (1



    Asymptotic behaviour of Hp (Rn×R+) (1

  2. Age-related decline in BubR1 impairs adult hippocampal neurogenesis

    Yang, Z; Jun, H.; Choi, C.I.; Yoo, K.H.; Cho, C.H.; Hussaini, S.M.; Simmons, A.J.; Kim, S.; Deursen, J.M.A. van; Baker, D.J.; Jang, M.H.


    Aging causes significant declines in adult hippocampal neurogenesis and leads to cognitive disability. Emerging evidence demonstrates that decline in the mitotic checkpoint kinase BubR1 level occurs with natural aging and induces progeroid features in both mice and children with mosaic variegated

  3. Scalar-tensor cosmology with R^{-1} curvature correction by Noether Symmetry

    Motavali, H; Jog, M Rowshan Almeh


    We discuss scalar-tensor cosmology with an extra $R^{-1}$ correction by the Noether Symmetry Approach. The existence of such a symmetry selects the forms of the coupling $\\omega(\\phi)$, of the potential $V(\\phi)$ and allows to obtain physically interesting exact cosmological solutions.

  4. Identifying the Proteins that Mediate the Ionizing Radiation Resistance of Deinococcus Radiodurans R1

    Battista, John R


    The primary objectives of this proposal was to define the subset of proteins required for the ionizing radiation (IR) resistance of Deinococcus radiodurans R1, characterize the activities of those proteins, and apply what was learned to problems of interest to the Department of Energy.

  5. R1 Resection by Necessity for Colorectal Liver Metastases Is It Still a Contraindication to Surgery?

    de Haas, Robbert J.; Wicherts, Dennis A.; Flores, Eduardo; Azoulay, Daniel; Castaing, Denis; Adam, Rene


    Objective: To compare long-term outcome of R0 (negative margins) and R1 (positive margins) liver resections for colorectal liver metastases (CLM) treated by an aggressive approach combining chemotherapy and repeat surgery. Summary Background Data: Complete macroscopic resection with negative margins

  6. Coordinated movement of cytoplasmic and transmembrane domains of RyR1 upon gating.

    Montserrat Samsó


    Full Text Available Ryanodine receptor type 1 (RyR1 produces spatially and temporally defined Ca2+ signals in several cell types. How signals received in the cytoplasmic domain are transmitted to the ion gate and how the channel gates are unknown. We used EGTA or neuroactive PCB 95 to stabilize the full closed or open states of RyR1. Single-channel measurements in the presence of FKBP12 indicate that PCB 95 inverts the thermodynamic stability of RyR1 and locks it in a long-lived open state whose unitary current is indistinguishable from the native open state. We analyzed two datasets of 15,625 and 18,527 frozen-hydrated RyR1-FKBP12 particles in the closed and open conformations, respectively, by cryo-electron microscopy. Their corresponding three-dimensional structures at 10.2 A resolution refine the structure surrounding the ion pathway previously identified in the closed conformation: two right-handed bundles emerging from the putative ion gate (the cytoplasmic "inner branches" and the transmembrane "inner helices". Furthermore, six of the identifiable transmembrane segments of RyR1 have similar organization to those of the mammalian Kv1.2 potassium channel. Upon gating, the distal cytoplasmic domains move towards the transmembrane domain while the central cytoplasmic domains move away from it, and also away from the 4-fold axis. Along the ion pathway, precise relocation of the inner helices and inner branches results in an approximately 4 A diameter increase of the ion gate. Whereas the inner helices of the K+ channels and of the RyR1 channel cross-correlate best with their corresponding open/closed states, the cytoplasmic inner branches, which are not observed in the K+ channels, appear to have at least as important a role as the inner helices for RyR1 gating. We propose a theoretical model whereby the inner helices, the inner branches, and the h1 densities together create an efficient novel gating mechanism for channel opening by relaxing two right

  7. Cdk5r1 Overexpression Induces Primary β-Cell Proliferation

    Carrie Draney


    Full Text Available Decreased β-cell mass is a hallmark of type 1 and type 2 diabetes. Islet transplantation as a method of diabetes therapy is hampered by the paucity of transplant ready islets. Understanding the pathways controlling islet proliferation may be used to increase functional β-cell mass through transplantation or by enhanced growth of endogenous β-cells. We have shown that the transcription factor Nkx6.1 induces β-cell proliferation by upregulating the orphan nuclear hormone receptors Nr4a1 and Nr4a3. Using expression analysis to define Nkx6.1-independent mechanisms by which Nr4a1 and Nr4a3 induce β-cell proliferation, we demonstrated that cyclin-dependent kinase 5 regulatory subunit 1 (Cdk5r1 is upregulated by Nr4a1 and Nr4a3 but not by Nkx6.1. Overexpression of Cdk5r1 is sufficient to induce primary rat β-cell proliferation while maintaining glucose stimulated insulin secretion. Overexpression of Cdk5r1 in β-cells confers protection against apoptosis induced by etoposide and thapsigargin, but not camptothecin. The Cdk5 kinase complex inhibitor roscovitine blocks islet proliferation, suggesting that Cdk5r1 mediated β-cell proliferation is a kinase dependent event. Overexpression of Cdk5r1 results in pRb phosphorylation, which is inhibited by roscovitine treatment. These data demonstrate that activation of the Cdk5 kinase complex is sufficient to induce β-cell proliferation while maintaining glucose stimulated insulin secretion.

  8. Neutrons characterization of the nuclear reactor Ian-R1 of Colombia; Caracterizacion de los neutrones del reactor nuclear IAN-R1 de Colombia

    Gonzalez P, L. X.; Martinez O, S. A. [Universidad Pedagogica y Tecnologica de Colombia, Grupo de Fisica Nuclear Aplicada y Simulacion, Carretera Central del Norte Km. 1, Via Paipa, 150003 Tunja, Boyaca (Colombia); Vega C, H. R., E-mail: [Universidad Autonoma de Zacatecas, Unidad Academica de Estudios Nucleares, Cipres No. 10, Fracc. La Penuela, 98068 Zacatecas (Mexico)


    By means of Monte Carlo methods, with the code MCNPX, the neutron characteristics of the research nuclear reactor Ian-R1 of Colombia, in power off but with the neutrons source in their start position, have been valued. The neutrons spectra, the total flow and their average power were calculated in the irradiation spaces inside the graphite reflector, as well as in the cells with air. Also the spectra, the total flow and the absorbed dose were calculated in several places distributed along the radial shaft inside the water moderator. The neutrons total flow was also considered to the long of the axial shaft. The characteristics of the neutrons spectra vary depending on their position regarding the source and the material that surrounds to the cell where the calculation was made. (Author)

  9. TRAPPIST monitoring of comets C/2012 S1 (ISON) and C/2013 R1 (Lovejoy)

    Opitom, C.; Jehin, E.; Manfroid, J.; Hutsemékers, D.; Gillon, M.


    We present the results of a dense photometric monitoring of comets C/2012 S1 (Ison) and C/2013 R1 (Lovejoy) using narrow-band cometary filters and the 60-cm TRAPPIST robotic telescope [1]. We were able to isolate the emission of the OH, NH, CN, C_2, and C_3 radicals for both comets as well as the dust continuum in four bands. By applying a Haser model [2] and fitting the observed profiles, we derive gas production rates. From the continuum bands, we computed the dust Afρ parameters [3]. We were able to follow the evolution of the gas and dust activity of these comets for weeks, looking for changes with the heliocentric distance, study the coma morphology, and analyze their composition and dust coma properties. Comet C/2012 S1 (ISON) was observed about three times a week from October 12 (r=1.43 au) to November 23, 2013. It was then at a heliocentric distance of 0.33 au, only five days before perihelion, when it disintegrated. This dense monitoring allowed us to detect fast changes of the cometary activity. We observed a slowly rising activity in October and early November, and two major outbursts around November 13 and November 19 [4], the gas and dust production rates being multiplied by at least a factor of five during each outburst and then slowly decreasing in the following days. These outbursts were correlated with changes in gas-production-rate ratios. The coma morphology study revealed strong jets in both gas and dust filters. Since the comet was very active in November, we were even able to detect OH jets in our images. Comet C/2013 R1 (Lovejoy) was observed before perihelion from September 9 (r=1.94 au) to November 16 (r=1.12 au), 2013 when the comet was too far North. We recovered the comet post-perihelion on February 13 (r=1.24 au), 2014 and planned to observe it until May (r=2.5 au) with narrow-band filters. We compare the evolution of gas and dust activity as well as the evolution of gas production rates ratios on both sides of perihelion. The

  10. Color-specific conditioning effects due to both orange and blue stimuli are observed in a Halobacterium salinarum strain devoid of putative methylatable sites on HtrI.

    Lucia, S; Cercignani, G; Frediani, A; Petracchi, D


    Behavioral responses of Halobacterium salinarum appear as changes in the frequency of motion reversals. Turning on orange light decreases the reversal frequency, whereas blue light induces reversals. Light pulses normally induce the same response as step-up stimuli. However, anomalous behavioral reactions, including inverse responses, are seen when stimuli are applied in sequence. The occurrence of a prior stimulus is conditioning for successive stimulation on a time scale of the same order of adaptational processes. These prolonged conditioning effects are color-specific. The only adaptation process identified so far is methylation of the transducers, and this could be somehow color-specific. Therefore we tested for the behavioral anomalies in a mutant in which all methylation sites on the transducer have been eliminated. The results show that behavioral anomalies are unaffected by the absence of methylation processes on the transducer.

  11. Characterization of AvaR1, a butenolide-autoregulator receptor for biosynthesis of a Streptomyces hormone in Streptomyces avermitilis.

    Sultan, Suandi Pratama; Kitani, Shigeru; Miyamoto, Kiyoko T; Iguchi, Hiroyuki; Atago, Tokitaka; Ikeda, Haruo; Nihira, Takuya


    Streptomyces hormones, sometimes called as autoregulators, are important signaling molecules to trigger secondary metabolism across many Streptomyces species. We recently identified a butenolide-type autoregulator (termed avenolide) as a new class of Streptomyces hormone from Streptomyces avermitilis that produces important anthelmintic agent avermectin. Avenolide triggers the production of avermectin with minimum effective concentration of nanomolar. Here, we describe the characterization of avaR1 encoding an avenolide receptor in the regulation of avermectin production and avenolide biosynthesis. The disruption of avaR1 resulted in transcriptional derepression of avenolide biosynthetic gene with an increase in avenolide production, with no change in the avermectin production profile. Moreover, the avaR1 mutant showed increased transcription of avaR1. Together with clear DNA-binding capacity of AvaR1 toward avaR1 upstream region, it suggests that AvaR1 negatively controls the expression of avaR1 through the direct binding to the promoter region of avaR1. These findings revealed that the avenolide receptor AvaR1 functions as a transcriptional repressor for avenolide biosynthesis and its own synthesis.

  12. The Indian origin of paternal haplogroup R1a1* substantiates the autochthonous origin of Brahmins and the caste system.

    Sharma, Swarkar; Rai, Ekta; Sharma, Prithviraj; Jena, Mamata; Singh, Shweta; Darvishi, Katayoon; Bhat, Audesh K; Bhanwer, A J S; Tiwari, Pramod Kumar; Bamezai, Rameshwar N K


    Many major rival models of the origin of the Hindu caste system co-exist despite extensive studies, each with associated genetic evidences. One of the major factors that has still kept the origin of the Indian caste system obscure is the unresolved question of the origin of Y-haplogroup R1a1*, at times associated with a male-mediated major genetic influx from Central Asia or Eurasia, which has contributed to the higher castes in India. Y-haplogroup R1a1* has a widespread distribution and high frequency across Eurasia, Central Asia and the Indian subcontinent, with scanty reports of its ancestral (R*, R1* and R1a*) and derived lineages (R1a1a, R1a1b and R1a1c). To resolve these issues, we screened 621 Y-chromosomes (of Brahmins occupying the upper-most caste position and schedule castes/tribals occupying the lower-most positions) with 55 Y-chromosomal binary markers and seven Y-microsatellite markers and compiled an extensive dataset of 2809 Y-chromosomes (681 Brahmins, and 2128 tribals and schedule castes) for conclusions. A peculiar observation of the highest frequency (up to 72.22%) of Y-haplogroup R1a1* in Brahmins hinted at its presence as a founder lineage for this caste group. Further, observation of R1a1* in different tribal population groups, existence of Y-haplogroup R1a* in ancestors and extended phylogenetic analyses of the pooled dataset of 530 Indians, 224 Pakistanis and 276 Central Asians and Eurasians bearing the R1a1* haplogroup supported the autochthonous origin of R1a1 lineage in India and a tribal link to Indian Brahmins. However, it is important to discover novel Y-chromosomal binary marker(s) for a higher resolution of R1a1* and confirm the present conclusions.

  13. PIK3R1 Mutations Cause Syndromic Insulin Resistance with Lipoatrophy

    Thauvin-Robinet, Christel; Auclair, Martine; Duplomb, Laurence; Caron-Debarle, Martine; Avila, Magali; St-Onge, Judith; Le Merrer, Martine; Le Luyer, Bernard; Héron, Delphine; Mathieu-Dramard, Michèle; Bitoun, Pierre; Petit, Jean-Michel; Odent, Sylvie; Amiel, Jeanne; Picot, Damien; Carmignac, Virginie; Thevenon, Julien; Callier, Patrick; Laville, Martine; Reznik, Yves; Fagour, Cédric; Nunes, Marie-Laure; Capeau, Jacqueline; Lascols, Olivier; Huet, Frédéric; Faivre, Laurence; Vigouroux, Corinne; Rivière, Jean-Baptiste


    Short stature, hyperextensibility of joints and/or inguinal hernia, ocular depression, Rieger anomaly, and teething delay (SHORT) syndrome is a developmental disorder with an unknown genetic cause and hallmarks that include insulin resistance and lack of subcutaneous fat. We ascertained two unrelated individuals with SHORT syndrome, hypothesized that the observed phenotype was most likely due to de novo mutations in the same gene, and performed whole-exome sequencing in the two probands and their unaffected parents. We then confirmed our initial observations in four other subjects with SHORT syndrome from three families, as well as 14 unrelated subjects presenting with syndromic insulin resistance and/or generalized lipoatrophy associated with dysmorphic features and growth retardation. Overall, we identified in nine affected individuals from eight families de novo or inherited PIK3R1 mutations, including a mutational hotspot (c.1945C>T [p.Arg649Trp]) present in four families. PIK3R1 encodes the p85α, p55α, and p50α regulatory subunits of class IA phosphatidylinositol 3 kinases (PI3Ks), which are known to play a key role in insulin signaling. Functional data from fibroblasts derived from individuals with PIK3R1 mutations showed severe insulin resistance for both proximal and distal PI3K-dependent signaling. Our findings extend the genetic causes of severe insulin-resistance syndromes and provide important information with respect to the function of PIK3R1 in normal development and its role in human diseases, including growth delay, Rieger anomaly and other ocular affections, insulin resistance, diabetes, paucity of fat, and ovarian cysts. PMID:23810378

  14. Improvement of Biodesulfurization Rate of Alginate Immobilized Rhodococcus erythropolis R1

    Derikvand, Peyman; Etemadifar, Zahra


    Background: Sulfur oxides released from the burning of oil causes severe environmental pollution. The sulfur can be removed via the 4S pathway in biodesulfurization (BDS). Immobilization approaches have been developed to prevent cell contamination of oil during the BDS process. Objectives: The encapsulation of Rhodococcus erythropolis R1 in calcium alginate beads was studied in order to enhance conversion of dibenzothiophene (DBT) to 2-hydroxy biphenyl (2-HBP) as the final product. Also the e...

  15. Insulin resistance uncoupled from dyslipidemia due to C-terminal PIK3R1 mutations

    Huang-Doran, Isabel; Tomlinson, Patsy; Payne, Felicity; Gast, Alexandra; Sleigh, Alison; Bottomley, William; Harris, Julie; Daly, Allan; Rocha, Nuno; Rudge, Simon; Clark, Jonathan; Kwok, Albert; Romeo, Stefano; McCann, Emma; Müksch, Barbara; Dattani, Mehul; Zucchini, Stefano; Wakelam, Michael; Foukas, Lazaros C.; Savage, David B.; Murphy, Rinki; O’Rahilly, Stephen; Semple, Robert K.


    Obesity-related insulin resistance is associated with fatty liver, dyslipidemia, and low plasma adiponectin. Insulin resistance due to insulin receptor (INSR) dysfunction is associated with none of these, but when due to dysfunction of the downstream kinase AKT2 phenocopies obesity-related insulin resistance. We report 5 patients with SHORT syndrome and C-terminal mutations in PIK3R1, encoding the p85α/p55α/p50α subunits of PI3K, which act between INSR and AKT in insulin signaling. Four of 5 patients had extreme insulin resistance without dyslipidemia or hepatic steatosis. In 3 of these 4, plasma adiponectin was preserved, as in insulin receptor dysfunction. The fourth patient and her healthy mother had low plasma adiponectin associated with a potentially novel mutation, p.Asp231Ala, in adiponectin itself. Cells studied from one patient with the p.Tyr657X PIK3R1 mutation expressed abundant truncated PIK3R1 products and showed severely reduced insulin-stimulated association of mutant but not WT p85α with IRS1, but normal downstream signaling. In 3T3-L1 preadipocytes, mutant p85α overexpression attenuated insulin-induced AKT phosphorylation and adipocyte differentiation. Thus, PIK3R1 C-terminal mutations impair insulin signaling only in some cellular contexts and produce a subphenotype of insulin resistance resembling INSR dysfunction but unlike AKT2 dysfunction, implicating PI3K in the pathogenesis of key components of the metabolic syndrome. PMID:27766312

  16. Core calculations for the upgrading of the IEA-R1 research reactor

    Santos, Adimir dos; Perrotta, Jose A.; Bastos, Jose Luis F.; Yamaguchi, Mitsuo; Umbehaun, Pedro E. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail:;;


    The IEA-R1 Research Reactor is a multipurpose reactor. It has been used for basic and applied research in the nuclear area, training and radioisotopes production since 1957. In 1995, the Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP) took the decision to modernize and upgrade the power from 2 to 5 MW and increase the operational cycle. This work presents the design requirements and the calculations effectuated to reach this goal. (author)

  17. Effects of heavy ions on inactivation and DNA double strand breaks in Deinococcus radiodurans R1.

    Zimmermann, H; Schafer, M; Schmitz, C; Bucker, H


    Inactivation and double strand break (dsb) induction after heavy ion irradiation were studied in stationary phase cells of the highly radiation resistant bacterium Deinococcus radiodurans R1. There is evidence that the radiation sensitivity of this bacterium is nearly independent on energy in the range of up to 15 MeV/u for lighter ions (Ar). The responses to dsb induction for charged particles show direct relationship between increasing radiation dose and residual intact DNA.

  18. AHR prevents human IL-1R1hi ILC3 differentiation to natural killer cells

    Hughes, Tiffany; Briercheck, Edward L.; Freud, Aharon G.; Trotta, Rossana; McClory, Susan; Scoville, Steven D.; Keller, Karen; Deng, Youcai; Cole, Jordan; Harrison, Nicholas; Mao, Charlene; Zhang, Jianying; Benson, Don M.; Yu, Jianhua; Caligiuri, Michael A.


    SUMMARY Accumulating evidence indicates that human natural killer (NK) cells develop in secondary lymphoid tissue (SLT) through a so-called “stage 3” developmental intermediate minimally characterized by a CD34-CD117+CD94- immunophenotype that lacks mature NK cell function. This stage 3 population is heterogeneous, potentially composed of functionally distinct innate lymphoid cell (ILC) types that includes interleukin-1 receptor (IL-1R1) positive, IL-22-producing ILC3s. Whether human ILC3s are developmentally related to NK cells is a subject of ongoing investigation. Here we show that antagonism of the aryl hydrocarbon receptor (AHR) or silencing of AHR gene expression promotes differentiation of tonsillar IL-22-producing IL-1R1hi human ILC3s to CD56brightCD94+ IFN-gamma-producing cytolytic mature NK cells expressing eomesodermin (EOMES) and T-Box Protein 21 (TBX21 or TBET). Hence, AHR is a transcription factor that prevents human IL-1R1hi ILC3s from differentiating into NK cells. PMID:24953655

  19. HCN observations of comets C/2013 R1 (Lovejoy) and C/2014 Q2 (Lovejoy)

    Wirström, E S; Källström, P; Levinsson, A; Olivefors, A; Tegehall, E


    HCN J=1-0 emission from the long-period comet C/2013 R1 (Lovejoy) was observed from the Onsala Space Observatory on multiple occasions during the month before its perihelion passage on December 22, 2013. We report detections for seven different dates, spanning heliocentric distances (R_h) decreasing from 0.94 to 0.82 au. Estimated HCN production rates are generally higher than previously reported for the same time period, but the implied increase in production rate with heliocentric distance, Q_{HCN} proportionate to R_h^{-3.2}, represent well the overall documented increase since it was first observed at R_h=1.35. The implied mean HCN abundance relative to water in R1 Lovejoy is 0.2%. We also report on a detection of HCN with the new 3 mm receiver system at Onsala Space Observatory in comet C/2014 Q2 (Lovejoy) on January 14, 2015, when its heliocentric distance was 1.3 au. Relative to comet C/2013 R1 (Lovejoy), the HCN production rate of C/2014 Q2 (Lovejoy) was more than 5 times higher at similar heliocentri...

  20. An optimized method for (15)N R(1) relaxation rate measurements in non-deuterated proteins.

    Gairí, Margarida; Dyachenko, Andrey; González, M Teresa; Feliz, Miguel; Pons, Miquel; Giralt, Ernest


    (15)N longitudinal relaxation rates are extensively used for the characterization of protein dynamics; however, their accurate measurement is hindered by systematic errors. (15)N CSA/(1)H-(15)N dipolar cross-correlated relaxation (CC) and amide proton exchange saturation transfer from water protons are the two main sources of systematic errors in the determination of (15)N R1 rates through (1)H-(15)N HSQC-based experiments. CC is usually suppressed through a train of 180° proton pulses applied during the variable (15)N relaxation period (T), which can perturb water magnetization. Thus CC cancellation is required in such a way as to minimize water saturation effects. Here we examined the level of water saturation during the T period caused by various types of inversion proton pulses to suppress CC: (I) amide-selective IBURP-2; (II) cosine-modulated IBURP-2; (III) Watergate-like blocks; and (IV) non-selective hard. We additionally demonstrate the effect of uncontrolled saturation of aliphatic protons on (15)N R1 rates. In this paper we present an optimized pulse sequence that takes into account the crucial effect of controlling also the saturation of the aliphatic protons during (15)N R1 measurements in non-deuterated proteins. We show that using cosine-modulated IBURP-2 pulses spaced 40 ms to cancel CC in this optimized pulse program is the method of choice to minimize systematic errors coming from water and aliphatic protons saturation effects.

  1. Porcine retinal cell line VIDO R1 and Chlamydia suis to modelize ocular chlamydiosis.

    Käser, Tobias; Cnudde, Thomas; Hamonic, Glenn; Rieder, Meghanne; Pasternak, J Alex; Lai, Ken; Tikoo, Suresh K; Wilson, Heather L; Meurens, François


    Human ocular Chlamydia trachomatis infections can lead to trachoma, the major cause of infectious blindness worldwide. Trachoma control strategies are very helpful but logistically challenging, and a trachoma vaccine is needed but not available. Pigs are a valuable large animal model for various immunological questions and could facilitate the study of human ocular chlamydial infections. In addition, a recent study identified the zoonotic potential of Chlamydia suis, the natural pathogen of pigs. In terms of the One Health Initiative, understanding the host-pathogen-interactions and finding a vaccine for porcine chlamydia infections would also benefit human health. Thus, we infected the porcine retinal cell line VIDO R1 with C. suis and analyzed the chlamydial life cycle and the innate immune response of the infected cells. Our results indicate that C. suis completes its life cycle in VIDO R1 cells within 48 h, comparable to C. trachomatis in humans. C. suis infection of VIDO R1 cells led to increased levels of various innate immune mediators like pathogen recognition receptors, cytokines and chemokines including IL6, TNFα, and MMP9, also most relevant in human C. trachomatis infections. These results illustrate the first steps in the host-pathogen-interactions of ocular C. suis infections in pigs and show their similarity to C. trachomatis infections in humans, justifying further testing of pigs as an animal model for human trachoma.

  2. Study of neutronic flux in IPR-R1 reactor with MCNPX; Estudo do fluxo neutronico no reator IPR-R1 com o MCNPX

    Melo, J.A.S.; Castrillo, L.S., E-mail:, E-mail: [Universidade de Pernambuco (UPE), Recife, PE (Brazil). Escola Politecnica; Oliveira, R.M.B.M., E-mail: [Secretaria Executiva de Educacao do Estado de Pernambuco (SEE), Recife, PE (Brazil)


    MCNPX computer code, one of the latest versions of code MCNP transport were used to study the flux distribution and its neutronic fluence as a function of energy in two research reactor irradiation IPR-R1. The model developed was validated with research conducted by Dalle (2005). Initially, in the simulation is considered fresh fuel whose core configuration contained three neutron rods control, being two of them 100% ejected while the other inserted 3,1 x 10{sup -1} m deep, as adopted in the literature situation. The neutron source used was the critical type, through KSRC card. The results of the neutron flow and neutronic fluence were obtained in the central tube and the turntable on a range of energy spectrum that ranged from 1.0 x 10{sup -9} MeV to 10 MeV, showing good correlations with the model used in validation. Finally, a hypothetical situation wherein the three reactor control rods are ejected simultaneously was simulated. The simulation results showed an increase in the neutron flux of 7% in the central tube and 5% on the turntable.

  3. MiR-23-TrxR1 as a novel molecular axis in skeletal muscle differentiation.

    Mercatelli, Neri; Fittipaldi, Simona; De Paola, Elisa; Dimauro, Ivan; Paronetto, Maria Paola; Jackson, Malcolm J; Caporossi, Daniela


    Thioredoxin reductase 1 (TrxR1) is a selenocysteine-containing protein involved in cellular redox homeostasis which is downregulated in skeletal muscle differentiation. Here we show that TrxR1 decrease occurring during myogenesis is functionally involved in the coordination of this cellular process. Indeed, TrxR1 depletion reduces myoblasts growth by inducing an early myogenesis -related gene expression pattern which includes myogenin and Myf5 up-regulation and Cyclin D1 decrease. On the contrary, the overexpression of TrxR1 during differentiation delays myogenic process, by negatively affecting the expression of Myogenin and MyHC. Moreover, we found that miR-23a and miR-23b - whose expression was increased in the early stage of C2C12 differentiation - are involved in the regulation of TrxR1 expression through their direct binding to the 3' UTR of TrxR1 mRNA. Interestingly, the forced inhibition of miR-23a and miR-23b during C2C12 differentiation partially rescues TrxR1 levels and delays the expression of myogenic markers, suggesting the involvement of miR-23 in myogenesis via TrxR1 repression. Taken together, our results depict for the first time a novel molecular axis, which functionally acts in skeletal muscle differentiation through the modulation of TrxR1 by miR-23.

  4. Genome-Wide Analysis of the Role of Global Transcriptional Regulator GntR1 in Corynebacterium glutamicum

    Tanaka, Yuya; Takemoto, Norihiko; Ito, Terukazu; Teramoto, Haruhiko; Yukawa, Hideaki; Inui, Masayuki


    The transcriptional regulator GntR1 downregulates the genes for gluconate catabolism and pentose phosphate pathway in Corynebacterium glutamicum. Gluconate lowers the DNA binding affinity of GntR1, which is probably the mechanism of gluconate-dependent induction of these genes. In addition, GntR1 positively regulates ptsG, a gene encoding a major glucose transporter, and pck, a gene encoding phosphoenolpyruvate carboxykinase. Here, we searched for the new target of GntR1 on a genome-wide scal...

  5. Association of social defeat stress-induced anhedonia-like symptoms with mGluR1-dependent decrease in membrane-bound AMPA-GluR1 in the mouse ventral midbrain.

    Yashiro, Sayori; Seki, Kenjiro


    Anhedonia is a core symptom of social defeat stress (SDS)-induced depression associated with the reward system. We previously reported that decreased membrane-bound AMPA-GluR1 in the reward system is associated with lipopolysaccharide-induced anhedonia-like symptoms. Since group I metabotropic glutamate receptor (mGluR) activation reduces the surface density of GluR1, we examined whether group I mGluR-dependent decrease in membrane-bound GluR1 in the reward system is involved in SDS-induced anhedonia-like symptoms. Mice exposed to SDS for 4 consecutive days had markedly decreased membrane-bound GluR1 and GluR2 in the prefrontal cortex (PFC) and membrane-bound GluR1 in the ventral midbrain (VM) along with lower sucrose preference (SP). Intra-PFC injection of the group I mGluR agonist (S)-3,5-dihydroxyphenylglycine (DHPG; 100 μmol) demonstrated decrease in membrane-bound GluR1 and GluR2 in the PFC 2 and 24 h and membrane-bound GluR1 in the VM 24 h after injection. Moreover, intra-PFC injection of DHPG decreased SP only in the second 24-h (24-48 h) period. Conversely, intra-VM injection of DHPG decreased SP in both the first and second 24-h period and decreased membrane-bound GluR1 in the VM 2 and 24 h after injection. Pre-treatment with the mGluR1 antagonist JNJ16259685 (30 mg/kg, subcutaneous) prevented SDS-decreased SP and membrane-bound GluR1 in the VM. The mGluR5 antagonist 2-methyl-6-(phenylethynyl)pyridine (MPEP; 10 mg/kg, subcutaneous) prevented SDS-induced decrease in membrane-bound GluR1 and GluR2 in the PFC, whereas MPEP did not affect SDS-induced decrease in SP and membrane-bound GluR1 in the VM. These results suggest that mGluR1-mediated decrease in membrane-bound GluR1 in VM is involved in SDS-induced anhedonia-like symptoms.

  6. The Q Motif Is Involved in DNA Binding but Not ATP Binding in ChlR1 Helicase.

    Hao Ding

    Full Text Available Helicases are molecular motors that couple the energy of ATP hydrolysis to the unwinding of structured DNA or RNA and chromatin remodeling. The conversion of energy derived from ATP hydrolysis into unwinding and remodeling is coordinated by seven sequence motifs (I, Ia, II, III, IV, V, and VI. The Q motif, consisting of nine amino acids (GFXXPXPIQ with an invariant glutamine (Q residue, has been identified in some, but not all helicases. Compared to the seven well-recognized conserved helicase motifs, the role of the Q motif is less acknowledged. Mutations in the human ChlR1 (DDX11 gene are associated with a unique genetic disorder known as Warsaw Breakage Syndrome, which is characterized by cellular defects in genome maintenance. To examine the roles of the Q motif in ChlR1 helicase, we performed site directed mutagenesis of glutamine to alanine at residue 23 in the Q motif of ChlR1. ChlR1 recombinant protein was overexpressed and purified from HEK293T cells. ChlR1-Q23A mutant abolished the helicase activity of ChlR1 and displayed reduced DNA binding ability. The mutant showed impaired ATPase activity but normal ATP binding. A thermal shift assay revealed that ChlR1-Q23A has a melting point value similar to ChlR1-WT. Partial proteolysis mapping demonstrated that ChlR1-WT and Q23A have a similar globular structure, although some subtle conformational differences in these two proteins are evident. Finally, we found ChlR1 exists and functions as a monomer in solution, which is different from FANCJ, in which the Q motif is involved in protein dimerization. Taken together, our results suggest that the Q motif is involved in DNA binding but not ATP binding in ChlR1 helicase.

  7. BDNF activates mTOR to regulate GluR1 expression required for memory formation.

    Leandro Slipczuk

    Full Text Available BACKGROUND: The mammalian target of Rapamycin (mTOR kinase plays a key role in translational control of a subset of mRNAs through regulation of its initiation step. In neurons, mTOR is present at the synaptic region, where it modulates the activity-dependent expression of locally-translated proteins independently of mRNA synthesis. Indeed, mTOR is necessary for different forms of synaptic plasticity and long-term memory (LTM formation. However, little is known about the time course of mTOR activation and the extracellular signals governing this process or the identity of the proteins whose translation is regulated by this kinase, during mnemonic processing. METHODOLOGY/PRINCIPAL FINDINGS: Here we show that consolidation of inhibitory avoidance (IA LTM entails mTOR activation in the dorsal hippocampus at the moment of and 3 h after training and is associated with a rapid and rapamycin-sensitive increase in AMPA receptor GluR1 subunit expression, which was also blocked by intra-hippocampal delivery of GluR1 antisense oligonucleotides (ASO. In addition, we found that pre- or post-training administration of function-blocking anti-BDNF antibodies into dorsal CA1 hampered IA LTM retention, abolished the learning-induced biphasic activation of mTOR and its readout, p70S6K and blocked GluR1 expression, indicating that BDNF is an upstream factor controlling mTOR signaling during fear-memory consolidation. Interestingly, BDNF ASO hindered LTM retention only when given into dorsal CA1 1 h after but not 2 h before training, suggesting that BDNF controls the biphasic requirement of mTOR during LTM consolidation through different mechanisms: an early one involving BDNF already available at the moment of training, and a late one, happening around 3 h post-training that needs de novo synthesis of this neurotrophin. CONCLUSIONS/SIGNIFICANCE: IN CONCLUSION, OUR FINDINGS DEMONSTRATE THAT: 1 mTOR-mediated mRNA translation is required for memory consolidation during

  8. Proposed design for the PGAA facility at the TRIGA IPR-R1 research reactor

    Guerra, Bruno T.; Jacimovic, Radojko; Menezes, Maria Angela BC; Leal,Alexandre S.


    Background This work presents an initial proposed design of a Prompt Gamma Activation Analysis (PGAA) facility to be installed at the TRIGA IPR-R1, a 60 years old research reactor of the Centre of Development of Nuclear Technology (CDTN) in Brazil. The basic characteristics of the facility and the results of the neutron flux are presented and discussed. Findings The proposed design is based on a quasi vertical tube as a neutron guide from the reactor core, inside the reactor pool, 6 m below t...

  9. Draft Genome Sequence of Raoultella terrigena R1Gly, a Diazotrophic Endophyte.

    Schicklberger, M; Shapiro, N; Loqué, D; Woyke, T; Chakraborty, R


    Raoultella terrigena R1Gly is a diazotrophic endophyte isolated from surface-sterilized roots of Nicotiana tabacum. The whole-genome sequence was obtained to investigate the endophytic characteristics of this organism at the genetic level, as well as to compare this strain with its close relatives. To our knowledge, this is the first genome obtained from the Raoultella terrigena species and only the third genome from the Raoultella genus, after Raoultella ornitholytic and Raoultella planticola. This genome will provide a foundation for further comparative genomic, metagenomic, and functional studies of this genus.

  10. Distinct domains in Bub1 localize RZZ and BubR1 to kinetochores to regulate the checkpoint

    Zhang, Gang; Lischetti, Tiziana; Hayward, Daniel G;


    , we show that a distinct region in Bub1 mediates kinetochore localization of BubR1 through direct binding, but surprisingly removal of this region increases checkpoint strength. Our work thus uncovers how Bub1 coordinates checkpoint signalling by distinct domains for RZZ and BubR1 recruitment...

  11. Bacterial Mitosis: ParM of Plasmid R1 Moves Plasmid DNA by an Actin-like Insertional Polymerization Mechanism

    Møller-Jensen, Jakob; Borch, Jonas; Dam, Mette


    Bacterial DNA segregation takes place in an active and ordered fashion. In the case of Escherichia coli plasmid R1, the partitioning system (par) separates paired plasmid copies and moves them to opposite cell poles. Here we address the mechanism by which the three components of the R1 par system...

  12. Evaluation of the physical protection system of the IEA-R1 research reactor

    Vaz, Antonio C.A.; Conti, Thadeu das N., E-mail:, E-mail: [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    The '09/11' in New York and the accident at the Fukushima power plant are two events that served as worldwide reference to review some aspects of the Physical Protection System (PPS) in nuclear areas. The nuclear research reactor IEA-R1 has followed this new world order and improved the protection systems that are directly related to detection (CCTV, sensors, alarms, etc), delay (turnstile, gates, barriers, etc) and response (communication systems, response force, etc), for operation against malicious act, seeking always to avoid or minimize any possibility of threat, theft and sabotage. These actions were performed to prevent and to mitigate the consequence on the environment, economy and society from damages caused by natural hazard, as well. This study evaluates the PPS of the IEA-R1 regarding the weaknesses, strengths,and impacts of the changes resulting from the system implanted. The analyses were based on methodology developed by security experts from SANDIA National Laboratories in Texas - U.S.A, allowing the evaluation of the system through probabilistic and hypothetical analysis. (author)

  13. The Pseudomonas aeruginosa chemotaxis methyltransferase CheR1 impacts on bacterial surface sampling.

    Juliane Schmidt

    Full Text Available The characterization of factors contributing to the formation and development of surface-associated bacterial communities known as biofilms has become an area of intense interest since biofilms have a major impact on human health, the environment and industry. Various studies have demonstrated that motility, including swimming, swarming and twitching, seems to play an important role in the surface colonization and establishment of structured biofilms. Thereby, the impact of chemotaxis on biofilm formation has been less intensively studied. Pseudomonas aeruginosa has a very complex chemosensory system with two Che systems implicated in flagella-mediated motility. In this study, we demonstrate that the chemotaxis protein CheR1 is a methyltransferase that binds S-adenosylmethionine and transfers a methyl group from this methyl donor to the chemoreceptor PctA, an activity which can be stimulated by the attractant serine but not by glutamine. We furthermore demonstrate that CheR1 does not only play a role in flagella-mediated chemotaxis but that its activity is essential for the formation and maintenance of bacterial biofilm structures. We propose a model in which motility and chemotaxis impact on initial attachment processes, dispersion and reattachment and increase the efficiency and frequency of surface sampling in P. aeruginosa.

  14. Experimental distribution of coolant in the IPR-R1 Triga nuclear reactor core

    Mesquita, Amir Z., E-mail: amir@cdtn.b [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil). Servico de Tecnologia de Reatores; Palma, Daniel A.P., E-mail: [Comissao Nacional de Energia Nuclear (CNEN/RJ), Rio de Janeiro, RJ (Brazil); Costa, Antonella L.; Pereira, Claubia; Veloso, Maria A.F.; Reis, Patricia A.L., E-mail: claubia@nuclear.ufmg.b, E-mail: dora@nuclear.ufmg.b [Universidade Federal de Minas Gerais (DEN/UFMG), Belo Horizonte, MG (Brazil). Dept. de Engenharia Nuclear


    The IPR-R1 is a typical TRIGA Mark I light-water and open pool type reactor. The core has an annular configuration of six rings and is cooled by natural circulation. The core coolant channels extend from the bottom grid plate to the top grid plate. The cooling water flows through the holes in the bottom grid plate, passes through the lower unheated region of the element, flows upwards through the active region, passes through the upper unheated region, and finally leaves the channel through the differential area between a triangular spacer block on the top of the fuel element and a round hole in the grid. Direct measurement of the flow rate in a coolant channel is difficult because of the bulky size and low accuracy of flow meters. The flow rate through the channel may be determined indirectly from the heat balance across the channel using measurements of the water inlet and outlet temperatures. This paper presents the experiments performed in the IPR-R1 reactor to monitoring some thermo-hydraulic parameters in the core coolant channels, such as: the radial and axial temperature profile, temperature, velocity, mass flow rate, mass flux and Reynolds's number. Some results were compared with theoretical predictions, as it was expected the variables follow the power distribution (or neutron flux) in the core. (author)

  15. Teste dos relógios e R-1 - forma B: evidências de validade

    Noronha, Ana Paula


    Full Text Available Considerando a necessidade de construção de testes brasileiros, com análises apropriadas de parâmetros psicométricos, o presente estudo objetivou analisar as evidências de validade do R-1: Teste não-verbal de avaliação da inteligência, por meio da correlação desse instrumento com Teste dos Relógios. Foram estudados 68 alunos de cursos de Educação de Jovens e Adultos, com idade média de 34,12 e desvio padrão de 13,76. Os resultados indicaram que a correlação entre os dois instrumentos foi significativa e com uma magnitude que pode ser considerada boa (0,64, indicando que 41% da variância é comum a ambos os testes. Entretanto, não foi aceita a hipótese de que o Teste dos Relógios estaria mais correlacionado com o Fator 2 do R-1

  16. Characterization of RyR1-slow, a ryanodine receptor specific to slow-twitch skeletal muscle.

    Morrissette, J; Xu, L; Nelson, A; Meissner, G; Block, B A


    Two distinct skeletal muscle ryanodine receptors (RyR1s) are expressed in a fiber type-specific manner in fish skeletal muscle (11). In this study, we compare [(3)H]ryanodine binding and single channel activity of RyR1-slow from fish slow-twitch skeletal muscle with RyR1-fast and RyR3 isolated from fast-twitch skeletal muscle. Scatchard plots indicate that RyR1-slow has a lower affinity for [(3)H]ryanodine when compared with RyR1-fast. In single channel recordings, RyR1-slow and RyR1-fast had similar slope conductances. However, the maximum open probability (P(o)) of RyR1-slow was threefold less than the maximum P(o) of RyR1-fast. Single channel studies also revealed the presence of two populations of RyRs in tuna fast-twitch muscle (RyR1-fast and RyR3). RyR3 had the highest P(o) of all the RyR channels and displayed less inhibition at millimolar Ca(2+). The addition of 5 mM Mg-ATP or 2.5 mM beta, gamma-methyleneadenosine 5'-triphosphate (AMP-PCP) to the channels increased the P(o) and [(3)H]ryanodine binding of both RyR1s but also caused a shift in the Ca(2+) dependency curve of RyR1-slow such that Ca(2+)-dependent inactivation was attenuated. [(3)H]ryanodine binding data also showed that Mg(2+)-dependent inhibition of RyR1-slow was reduced in the presence of AMP-PCP. These results indicate differences in the physiological properties of RyRs in fish slow- and fast-twitch skeletal muscle, which may contribute to differences in the way intracellular Ca(2+) is regulated in these muscle types.

  17. Development of new plasmid DNA vaccine vectors with R1-based replicons

    Bower Diana M


    Full Text Available Abstract Background There has been renewed interest in biopharmaceuticals based on plasmid DNA (pDNA in recent years due to the approval of several veterinary DNA vaccines, on-going clinical trials of human pDNA-based therapies, and significant advances in adjuvants and delivery vehicles that have helped overcome earlier efficacy deficits. With this interest comes the need for high-yield, cost-effective manufacturing processes. To this end, vector engineering is one promising strategy to improve plasmid production. Results In this work, we have constructed a new DNA vaccine vector, pDMB02-GFP, containing the runaway R1 origin of replication. The runaway replication phenotype should result in plasmid copy number amplification after a temperature shift from 30°C to 42°C. However, using Escherichia coli DH5α as a host, we observed that the highest yields of pDMB02-GFP were achieved during constant-temperature culture at 30°C, with a maximum yield of approximately 19 mg pDNA/g DCW being observed. By measuring mRNA and protein levels of the R1 replication initiator protein, RepA, we determined that RepA may be limiting pDMB02-GFP yield at 42°C. A mutant plasmid, pDMB-ATG, was constructed by changing the repA start codon from the sub-optimal GTG to ATG. In cultures of DH5α[pDMB-ATG], temperature-induced plasmid amplification was more dramatic than that observed with pDMB02-GFP, and RepA protein was detectable for several hours longer than in cultures of pDMB02-GFP at 42°C. Conclusions Overall, we have demonstrated that R1-based plasmids can produce high yields of high-quality pDNA without the need for a temperature shift, and have laid the groundwork for further investigation of this class of vectors in the context of plasmid DNA production.

  18. A structural model of the pore-forming region of the skeletal muscle ryanodine receptor (RyR1.

    Srinivas Ramachandran


    Full Text Available Ryanodine receptors (RyRs are ion channels that regulate muscle contraction by releasing calcium ions from intracellular stores into the cytoplasm. Mutations in skeletal muscle RyR (RyR1 give rise to congenital diseases such as central core disease. The absence of high-resolution structures of RyR1 has limited our understanding of channel function and disease mechanisms at the molecular level. Here, we report a structural model of the pore-forming region of RyR1. Molecular dynamics simulations show high ion binding to putative pore residues D4899, E4900, D4938, and D4945, which are experimentally known to be critical for channel conductance and selectivity. We also observe preferential localization of Ca(2+ over K(+ in the selectivity filter of RyR1. Simulations of RyR1-D4899Q mutant show a loss of preference to Ca(2+ in the selectivity filter as seen experimentally. Electrophysiological experiments on a central core disease mutant, RyR1-G4898R, show constitutively open channels that conduct K(+ but not Ca(2+. Our simulations with G4898R likewise show a decrease in the preference of Ca(2+ over K(+ in the selectivity filter. Together, the computational and experimental results shed light on ion conductance and selectivity of RyR1 at an atomistic level.

  19. Temperature dependence of the chromium(III) R1 linewidth in emerald

    Carceller-Pastor, Ivana; Hutchison, Wayne D.; Riesen, Hans


    The temperature dependent contribution to the R1 (2E ← 4A2) linewidth in emerald, Be3Al2Si6O18:Cr3, has been measured by employing spectral hole-burning, fluorescence line narrowing and conventional luminescence experiments. The contribution varies from 0.6 MHz at 6.5 K to ˜420 GHz at 240 K and the line red-shifts by ˜570 GHz. Above 60 K, the dependence is well described by a non-perturbative formalism for two-phonon Raman scattering. Below this temperature the direct one-phonon process between the levels of the split 2E excited state dominates. However, it appears that a localized low-energy phonon leads to a deviation from the standard pattern at lowest temperatures.

  20. Structure determination of the neutral exopolysaccharide produced by Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1.

    Van Calsteren, Marie-Rose; Gagnon, Fleur; Nishimura, Junko; Makino, Seiya


    The neutral exopolysaccharide (NPS) of Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 was purified and characterized. The molecular mass was 5.0×10(6) g/mol. Sugar and absolute configuration analyses gave the following composition: d-Glc, 1; d-Gal, 1.5. The NPS was also submitted to periodate oxidation followed by borohydride reduction and Smith degradation. Sugar and methylation analyses, (1)H and (13)C nuclear magnetic resonance, and mass spectrometry of the NPS or of its specifically modified products allowed determining the repeating unit sequence: {2)Glc(α1-3)Glc(β1-3)[Gal(β1-4)]Gal(β1-4)Gal(α1-}n. The structure is compared to that of exopolysaccharides produced by other Lactobacillus bulgaricus strains.

  1. The interaction of the Eco R1 restriction enzyme E.coli with nucleotides

    Hollis, Donald F. [Univ. of California, Berkeley, CA (United States)


    The Eco R1 restriction enzyme can be shown to be inhibited by nucleotides which correspond to any part of its known site of phosphodiesterase activity. A series of di-, tetra-, and hexa-nucleotide fragments were synthesized and their effect on the activity of the enzyme upon superhelical Co1 E1 DNA studied. The inhibition caused by the individual mononucleotides were also studied. In general all the nucleotide fragments showed some form of interaction with the enzyme system. Tetranucleotides were stronger inhibitors than dinucleotides, which in turn were stronger inhibitors than the mononucleotides. Within each category of inhibitors, those containing the phosphodiester bond which is acted upon by the enzyme were the strongest inhibitors. Only those fragments which were consistent with the enzymes site of activity showed competitive inhibition kinetics. Nucleotides which do not fit within the site of phosphodiesterase activity show non-competitive inhibition kinetics.

  2. How chromosome mis-segregation leads to cancer: lessons from BubR1 mouse models.

    Lee, Hyunsook


    Alteration in chromosome numbers and structures instigate and foster massive genetic instability. As Boveri has seen a hundred years ago (Boveri, 1914; 2008), aneuploidy is hallmark of many cancers. However, whether aneuploidy is the cause or the result of cancer is still at debate. The molecular mechanism behind aneuploidy includes the chromo-some mis-segregation in mitosis by the compromise of spindle assembly checkpoint (SAC). SAC is an elaborate network of proteins, which monitor that all chromosomes are bipolarly attached with the spindles. Therefore, the weakening of the SAC is the major reason for chromosome number instability, while complete compromise of SAC results in detrimental death, exemplified in natural abortion in embryonic stage. Here, I will review on the recent progress on the understanding of chromosome mis-segregation and cancer, based on the comparison of different mouse models of BubR1, the core component of SAC.

  3. Harvesting biohydrogen from cellobiose from sulfide or nitrite-containing wastewaters using Clostridium sp. R1.

    Ho, Kuo-Ling; Lee, Duu-Jong


    Harvesting biohydrogen from inhibiting wastewaters is of practical interest since the toxicity of compounds in a wastewater stream commonly prevents the bioenergy content being recovered. The isolated Clostridium sp. R1 is utilized to degrade cellobiose in sulfide or nitrite-containing medium for biohydrogen production. The strain can effectively degrade cellobiose free of severe inhibitory effects at up to 200 mgl(-1) sulfide or to 5 mgl(-1) nitrite, yielding hydrogen at >2.0 mol H2 mol(-1) cellobiose. Principal metabolites of cellobiose fermentation are acetate and butyrate, with the concentration of the former increases with increasing sulfide and nitrite concentrations. The isolated strain can yield hydrogen from cellobiose in sulfide-laden wastewaters. However, the present of nitrite significantly limit the efficiency of the biohydrogen harvesting process.

  4. Role of miR-1 and miR-133a in myocardial ischemic postconditioning

    Xie Bing


    Full Text Available Abstract Background Ischemic postconditioning (IPost has aroused much attention since 2003 when it was firstly reported. The role of microRNAs (miRNAs or miRs in IPost has rarely been reported. The present study was undertaken to investigate whether miRNAs were involved in the protective effect of IPost against myocardial ischemia-reperfusion (IR injury and the probable mechanisms involved. Methods Thirty SD rats weighing 250-300 g were equally randomized to three groups: Control group, where the rats were treated with thoracotomy only; IR group, where the rats were treated with ischemia for 60 min and reperfusion for 180 min; and IPost group, where the rats were treated with 3 cycles of transient IR just before reperfusion. The extent of myocardial infarction, LDH and CK activities were measured immediately after treatment. Myocardial apoptosis was detected by TUNEL assay. The myocardial tissue was collected after IR or IPost stimulation to evaluate the miRNAs expression level by miRNA-microarray and quantitative real-time RT-PCR. Real-time PCR was conducted to identify changes in mRNA expression of apoptosis-related genes such as Bcl-2, Bax and Caspase-9 (CASP9, and Western blot was used to compare the protein expression level of CASP9 in the three groups. The miRNA mimics and anti-miRNA oligonucleotides (AMO were transferred into the cultured neonatal cardiomyocytes and myocardium before they were treated with IR. The effect of miRNAs on apoptosis was determined by flow cytometry and TUNEL assay. CASP9, as one of the candidate target of miR-133a, was compared during IR after the miR-133a mimic or AMO-133a was transferred into the myocardium. Results IPost reduced the IR-induced infarct size of the left ventricle, and decreased CK and LDH levels. TUNEL assay showed that myocardial apoptosis was attenuated by IPost compared with IR. MiRNA-microarray and RT-PCR showed that myocardial-specific miR-1 and miR-133a were down-regulated by IR, and up

  5. Optimization of the irradiation beam in the bnct research facility at IEA-R1 reactor

    Vinicius Alexandre de Castro


    A Terapia por Captura de Nêutrons pelo Boro (BNCT) é uma técnica radioterapêutica, que visa o tratamento de alguns tipos de câncer, em que sua energia útil é proveniente da reação nuclear promovida pela incidência de nêutrons térmicos no isótopo de 10B. No Brasil existe uma instalação, localizada junto ao canal de irradiação número 3 do Reator de Pesquisas IEA-R1 do IPEN, que foi projetada para o desenvolvimento de pesquisas em BNCT. Para uma aplicação adequada da técnica é necessário que o f...

  6. The role of TonB-dependent receptor TbdR1 in Riemerella anatipestifer in iron acquisition and virulence.

    Lu, Fengying; Miao, Shuang; Tu, Jing; Ni, Xintao; Xing, Linlin; Yu, Hui; Pan, Ling; Hu, Qinghai


    Riemerella anatipestifer is an important duck pathogen and causes serious economic losses to the duck industry worldwide. To date, four full R. anatipestifer genomic sequences have been submitted to the GenBank database and 31 TonB-dependent outer membrane receptors, which may play critical roles in host-bacteria interactions, were predicted for R. anatipestifer strain GSM15868. In our previous study, we reported that the TonB-dependent receptor TbdR1 was a cross immunogenic antigen among R. anatipestifer serotypes 1, 2, and 10. However, the biological functions of TbdR1 in R. anatipestifer remain unclear. In the present study, a tbdR1 (Riean_1607) deletion mutant CH3ΔtbdR1 of R. anatipestifer strain CH3 was constructed and characterized for iron-limited growth, biofilm formation, and pathogenicity to ducklings. Our results showed that TbdR1 was involved in hemin iron acquisition and the tbdR1 deletion significantly reduced biofilm formation and adhesion to and invasion of Vero cells. Animal experiments indicated that the median lethal dose of the CH3ΔtbdR1 mutant in ducklings was about 45-fold higher than that of the wild-type CH3 strain. Additional analysis indicated that bacterial loads in blood, liver, and brain tissues in CH3ΔtbdR1-infected ducklings were decreased significantly compared to those in wild-type CH3-infected ducklings. Thus, our results demonstrated that TbdR1 was involved in hemin iron acquisition and necessary for optimal bacterial virulence. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Virtual Screening and Molecular Dynamics Study of Potential Negative Allosteric Modulators of mGluR1 from Chinese Herbs

    Ludi Jiang


    Full Text Available The metabotropic glutamate subtype 1 (mGluR1, a member of the metabotropic glutamate receptors, is a therapeutic target for neurological disorders. However, due to the lower subtype selectivity of mGluR1 orthosteric compounds, a new targeted strategy, known as allosteric modulators research, is needed for the treatment of mGluR1-related diseases. Recently, the structure of the seven-transmembrane domain (7TMD of mGluR1 has been solved, which reveals the binding site of allosteric modulators and provides an opportunity for future subtype-selectivity drug design. In this study, a series of computer-aided drug design methods were utilized to discover potential mGluR1 negative allosteric modulators (NAMs. Pharmacophore models were constructed based on three different structure types of mGluR1 NAMs. After validation using the built-in parameters and test set, the optimal pharmacophore model of each structure type was selected and utilized as a query to screen the Traditional Chinese Medicine Database (TCMD. Then, three different hit lists of compounds were obtained. Molecular docking was used based on the latest crystal structure of mGluR1-7TMD to further filter these hits. As a compound with high QFIT and LibDock Score was preferred, a total of 30 compounds were retained. MD simulation was utilized to confirm the stability of potential compounds binding. From the computational results, thesinine-4ʹ-O-β-d-glucoside, nigrolineaxanthone-P and nodakenin might exhibit negative allosteric moderating effects on mGluR1. This paper indicates the applicability of molecular simulation technologies for discovering potential natural mGluR1 NAMs from Chinese herbs.

  8. How the nuclear safety team conducts emergency exercises at the IEA-R1 reactor

    Vaz, Antonio C.A.; Silva, Davilson G.; Toyoda, Eduardo Y.; Santia, Paulo S.; Conti, Thadeu N.; Semmler, Renato; Carvalho, Ricardo N., E-mail:, E-mail:, E-mail:, E-mail:, E-mail:, E-mail: rsemmler@ipen.b, E-mail: [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    This work introduces the Diagram of Emergency Exercise Coordination designed by the Nuclear Safety Team for better Emergency Exercise coordination. The Nuclear Safety Team was created with the mission of avoiding, preventing and mitigating the causes and effects of accidents at the IEA-R1. The facility where we conduct our work is located in an area of a huge population, what increases the responsibility of our mission: conducting exercises and training are part of our daily activities. During the Emergency Exercise, accidents ranked 0-4 on INES (International Nuclear Events Scale) are simulated and involve: Police Department, Fire Department, workers, people from the community, and others. In the last exercise held in June 2014, the scenario contemplated a terrorist organization action that infiltrated in a group of students who were visiting the IEA-R1, tried to steal fresh fuel element to fabricate a dirty bomb. Emergency procedures and plans, timeline and metrics of the actions were applied to the Emergency Exercise evaluation. The next exercise will be held in November, with the simulation of the piping of the primary cooling circuit rupture, causing the emptying of the pool and the lack of cooling of the fuel elements in the reactor core: this will be the scenario. The skills acquired and the systems improvement have been very important tools for the reactor operation safety and the Nuclear Safety Team is making technical efforts so that these Emergency Exercises may be applied to other nuclear and radiological facilities. Equally important for the process of improving nuclear safety is the emphasis placed on implementing quality improvements to the human factor in the nuclear safety area, a crucial element that is often not considered by those outside the nuclear sector. Surely, the Diagram of Emergency Exercise Coordination application will improve and facilitate the organization, coordination and evaluation tasks. (author)

  9. Protein conformational exchange measured by 1H R1ρ relaxation dispersion of methyl groups.

    Weininger, Ulrich; Blissing, Annica T; Hennig, Janosch; Ahlner, Alexandra; Liu, Zhihong; Vogel, Hans J; Akke, Mikael; Lundström, Patrik


    Activated dynamics plays a central role in protein function, where transitions between distinct conformations often underlie the switching between active and inactive states. The characteristic time scales of these transitions typically fall in the microsecond to millisecond range, which is amenable to investigations by NMR relaxation dispersion experiments. Processes at the faster end of this range are more challenging to study, because higher RF field strengths are required to achieve refocusing of the exchanging magnetization. Here we describe a rotating-frame relaxation dispersion experiment for (1)H spins in methyl (13)CHD2 groups, which improves the characterization of fast exchange processes. The influence of (1)H-(1)H rotating-frame nuclear Overhauser effects (ROE) is shown to be negligible, based on a comparison of R 1ρ relaxation data acquired with tilt angles of 90° and 35°, in which the ROE is maximal and minimal, respectively, and on samples containing different (1)H densities surrounding the monitored methyl groups. The method was applied to ubiquitin and the apo form of calmodulin. We find that ubiquitin does not exhibit any (1)H relaxation dispersion of its methyl groups at 10 or 25 °C. By contrast, calmodulin shows significant conformational exchange of the methionine methyl groups in its C-terminal domain, as previously demonstrated by (1)H and (13)C CPMG experiments. The present R 1ρ experiment extends the relaxation dispersion profile towards higher refocusing frequencies, which improves the definition of the exchange correlation time, compared to previous results.

  10. An Investigation of the Stoichiometry of Na+ Cotransport with Dopamine in Rat and Human Dopamine Transporters Expressed in Human Embryonic Kidney Cells


    fine-tuning of movement and the coloring of mood (Blakely and Bauman, 2000). Dopamine and the neuronal dopamine transporter (DAT) have been studied... Halobacterium halobium. Translocation stoichiometries and apparent cooperativity. Biochemistry 17, 3011-3018. McElvain J. S. and Schenk J. O. (1992) A

  11. A comparison of an ATPase from the archaebacterium Halobacterium saccharovorum with the F1 moiety from the Escherichia coli ATP Synthase

    Stan-Lotter, Helga; Hochstein, Lawrence I.


    A purified ATPase associated with membranes from Halobacterium saccharovorum was compared with the F sub 1 moiety from the Escherichia coli ATP Synthase. The halobacterial enzyme was composed of two major (I and II) and two minor subunits (III and IV), whose molecular masses were 87 kDa, 60 kDa, 29 kDa, and 20 kDa, respectively. The isoelectric points of these subunits ranged from 4.1 to 4.8, which in the case of the subunits I and II was consistent with the presence of an excess of acidic amino acids (20 to 22 Mol percent). Peptide mapping of sodium dodecylsulfate-denatured subunits I and II showed no relationship between the primary structures of the individual halobacterial subunits or similarities to the subunits of the F sub 1 ATPase (EC from E. coli. Trypsin inactivation of the halobacterial ATPase was accompanied by the partial degradation of the major subunits. This observation, taken in conjunction with molecular masses of the subunits and the native enzyme, was consistent with the previously proposed stoichiometry of 2:2:1:1. These results suggest that H. saccharovorum, and possibly, Halobacteria in general, possess an ATPase which is unlike the ubiquitous F sub o F sub 1 - ATP Synthase.

  12. NCoR1-independent mechanism plays a role in the action of the unliganded thyroid hormone receptor.

    Mendoza, Arturo; Astapova, Inna; Shimizu, Hiroaki; Gallop, Molly R; Al-Sowaimel, Lujain; MacGowan, S M Dileas; Bergmann, Tim; Berg, Anders H; Tenen, Danielle E; Jacobs, Christopher; Lyubetskaya, Anna; Tsai, Linus; Hollenberg, Anthony N


    Nuclear receptor corepressor 1 (NCoR1) is considered to be the major corepressor that mediates ligand-independent actions of the thyroid hormone receptor (TR) during development and in hypothyroidism. We tested this by expressing a hypomorphic NCoR1 allele (NCoR1ΔID), which cannot interact with the TR, in Pax8-KO mice, which make no thyroid hormone. Surprisingly, abrogation of NCoR1 function did not reverse the ligand-independent action of the TR on many gene targets and did not fully rescue the high mortality rate due to congenital hypothyroidism in these mice. To further examine NCoR1's role in repression by the unliganded TR, we deleted NCoR1 in the livers of euthyroid and hypothyroid mice and examined the effects on gene expression and enhancer activity measured by histone 3 lysine 27 (H3K27) acetylation. Even in the absence of NCoR1 function, we observed strong repression of more than 43% of positive T3 (3,3',5-triiodothyronine) targets in hypothyroid mice. Regulation of approximately half of those genes correlated with decreased H3K27 acetylation, and nearly 80% of these regions with affected H3K27 acetylation contained a bona fide TRβ1-binding site. Moreover, using liver-specific TRβ1-KO mice, we demonstrate that hypothyroidism-associated changes in gene expression and histone acetylation require TRβ1. Thus, many of the genomic changes mediated by the TR in hypothyroidism are independent of NCoR1, suggesting a role for additional signaling modulators in hypothyroidism.

  13. Impaired associative fear learning in mice with complete loss or haploinsufficiency of AMPA GluR1 receptors

    Michael Feyder


    Full Text Available There is compelling evidence that L-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA glutamate receptors containing the GluR1 subunit contribute to the molecular mechanisms associated with learning. AMPA GluR1 glutamate receptor knockout mice (KO exhibit abnormal hippocampal and amygdala plasticity, and deficits on various assays for cognition including Pavlovian fear conditioning. Here we examined associative fear learning in mice with complete absence (KO or partial loss (heterozygous mutant, HET of GluR1 on multiple fear conditioning paradigms. After multi-trial delay or trace conditioning, KO displayed impaired tone and context fear recall relative to WT, whereas HET were normal. After one-trial delay conditioning, both KO and HET showed impaired tone and context recall. HET and KO showed normal nociceptive sensitivity in the hot plate and tail flick tests. These data demonstrate that the complete absence of GluR1 subunit-containing receptors prevents the formation of associative fear memories, while GluR1 haploinsufficiency is sufficient to impair one-trial fear learning. These findings support growing evidence of a major role for GluR1-containing AMPA receptors in amygdalamediated forms of learning and memory.

  14. ERp46 binds to AdipoR1, but not AdipoR2, and modulates adiponectin signalling

    Charlton, Hayley K.; Webster, Julie; Kruger, Sarah; Simpson, Fiona; Richards, Ayanthi A. [Diamantina Institute for Cancer, Immunology and Metabolic Medicine, University of Queensland, Princess Alexandra Hospital, Brisbane, QLD 4102 (Australia); Whitehead, Jonathan P., E-mail: [Diamantina Institute for Cancer, Immunology and Metabolic Medicine, University of Queensland, Princess Alexandra Hospital, Brisbane, QLD 4102 (Australia)


    The pleiotropic effects of the insulin-sensitizing adipokine adiponectin are mediated, at least in part, by two seven-transmembrane domain receptors AdipoR1 and AdipoR2. Recent reports indicate a role for AdipoR-binding proteins, namely APPL1, RACK1 and CK2{beta}, in proximal signal transduction events. Here we demonstrate that endoplasmic reticulum protein 46 (ERp46) interacts specifically with AdipoR1 and provide evidence that ERp46 modulates adiponectin signalling. Co-immunoprecipitation followed by mass spectrometry identified ERp46 as an AdipoR1-, but not AdipoR2-, interacting protein. Analysis of truncated constructs and GST-fusion proteins revealed the interaction was mediated by the cytoplasmic, N-terminal residues (1-70) of AdipoR1. Indirect immunofluorescence microscopy and subcellular fractionation studies demonstrated that ERp46 was present in the ER and the plasma membrane (PM). Transient knockdown of ERp46 increased the levels of AdipoR1, and AdipoR2, at the PM and this correlated with increased adiponectin-stimulated phosphorylation of AMPK. In contrast, adiponectin-stimulated phosphorylation of p38MAPK was reduced following ERp46 knockdown. Collectively these results establish ERp46 as the first AdipoR1-specific interacting protein and suggest a role for ERp46 in adiponectin receptor biology and adiponectin signalling.

  15. Mouse oocytes depend on BubR1 for proper chromosome segregation but not for prophase I arrest

    Touati, Sandra A.; Buffin, Eulalie; Cladière, Damien; Hached, Khaled; Rachez, Christophe; van Deursen, Jan M.; Wassmann, Katja


    Mammalian female meiosis is error prone, with rates of meiotic chromosome missegregations strongly increasing towards the end of the reproductive lifespan. A strong reduction of BubR1 has been observed in oocytes of women approaching menopause and in ovaries of aged mice, which led to the hypothesis that a gradual decline of BubR1 contributes to age-related aneuploidization. Here we employ a conditional knockout approach in mouse oocytes to dissect the meiotic roles of BubR1. We show that BubR1 is required for diverse meiotic functions, including persistent spindle assembly checkpoint activity, timing of meiosis I and the establishment of robust kinetochore-microtubule attachments in a meiosis-specific manner, but not prophase I arrest. These data reveal that BubR1 plays a multifaceted role in chromosome segregation during the first meiotic division and suggest that age-related decline of BubR1 is a key determinant of the formation of aneuploid oocytes as women approach menopause. PMID:25897860

  16. Deacetylation of the mitotic checkpoint protein BubR1 at lysine 250 by SIRT2 and subsequent effects on BubR1 degradation during the prometaphase/anaphase transition.

    Suematsu, Tomohisa; Li, Yanze; Kojima, Hirotada; Nakajima, Koichi; Oshimura, Mitsuo; Inoue, Toshiaki


    Mitotic catastrophe, a form of cell death that occurs during mitosis and after mitotic slippage to a tetraploid state, plays an important role in the efficacy of cancer cell killing by microtubule inhibitors. Prolonged mitotic arrest at the spindle assembly checkpoint (SAC) is a well-known requirement for mitotic catastrophe and, thus, for conferring sensitivity to microtubule inhibitors. We previously reported that downregulation of SIRT2, a member of the sirtuin family of NAD+-dependent deacetylases, confers resistance to microtubule inhibitors by abnormally prolonging mitotic arrest and thus compromising the cell death pathway after mitotic slippage. Thus, turning off SAC activation after a defined period is an additional requirement for efficient post-slippage death. Here, we investigated whether SIRT2 deacetylates BubR1, which is a core component of the SAC; acetylation of BubR1 at lysine 250 (K250) during prometaphase inhibits its APC/C-dependent proteolysis and thus regulates timing in anaphase entry. We showed that SIRT2 deacetylates BubR1 K250 both in vitro and in vivo. We also found that SIRT2 knockdown leads to increased levels of BubR1 acetylation at prometaphase; however, this increase is not substantial to elevate the levels of total BubR1 or delay the transition from prometaphase to anaphase. The present study shows that SIRT2 is a deacetylase for BubR1 K250, although the abnormally prolonged SAC activation observed in SIRT2 knockdown cells is not accompanied by a change in BubR1 levels or by delayed progression from prometaphase to anaphase.

  17. Spent fuel management - two alternatives at the FiR 1 reactor

    Salmenhaara, S.E.J. [Technical Research Centre of Finland (VTT), FIN-02044 VTT Espoo (Finland)


    The FiR 1 -reactor, a 250 kW Triga reactor, has been in operation since 1962. The reactor with its subsystems has experienced a large renovation work in 1996-97. The main purpose of the upgrading was to install the new Boron Neutron Capture Therapy (BNCT) irradiation facility. The BNCT work dominates the current utilization of the reactor: four days per week for BNCT purposes and only one day per week for neutron activation analysis and isotope production. The Council of State (government) granted for the reactor a new operating license for twelve years starting from the beginning of the year 2000. There is however a special condition in the new license. We have to achieve a binding agreement between our Research Centre and the domestic Nuclear Power Plant Companies about the possibility to use the final disposal facility of the Nuclear Power Plants for our spent fuel, if we want to continue the reactor operation beyond the year 2006. In addition to the choosing of one of the spent fuel management alternatives the future of the reactor will also depend strongly on the development of the BNCT irradiations. If the number of patients per year increases fast enough and the irradiations of the patients will be economically justified, the operation of the reactor will continue independently of the closing of the USDOE alternative in 2006. Otherwise, if the number of patients will be low, the funding of the reactor will be probably stopped and the reactor will be shut down. (author)

  18. Mechanism of DNA Segregation in Prokaryotes: Replicon Pairing by parC of Plasmid R1

    Jensen, Rasmus Bugge; Lurz, Rudi; Gerdes, Kenn


    Prokaryotic chromosomes and plasmids encode partitioning systems that are required for DNA segregation at cell division. The systems are thought to be functionally analogous to eukaryotic centromeres and to play a general role in DNA segregation. The parA system of plasmid R1 encodes two proteins ParM and ParR, and a cis-acting centromere-like site denoted parC. The ParR protein binds to parC in vivo and in vitro. The ParM protein is an ATPase that interacts with ParR specifically bound to parC. Using electron microscopy, we show here that parC mediates efficient pairing of plasmid molecules. The pairing requires binding of ParR to parC and is stimulated by the ParM ATPase. The ParM mediated stimulation of plasmid pairing is dependent on ATP hydrolysis by ParM. Using a ligation kinetics assay, we find that ParR stimulates ligation of parC-containing DNA fragments. The rate-of-ligation was increased by wild type ParM protein but not by mutant ParM protein deficient in the ATPase activity. Thus, two independent assays show that parC mediates pairing of plasmid molecules in vitro. These results are consistent with the proposal that replicon pairing is part of the mechanism of DNA segregation in prokaryotes.

  19. Measured and calculated effective delayed neutron fraction of the IPR-R1 Triga reactor

    Souza, Rose Mary G.P.; Dalle, Hugo M.; Campolina, Daniel A.M., E-mail: souzarm@cdtn.b, E-mail: dallehm@cdtn.b, E-mail: campolina@cdtn.b [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)


    The effective delayed neutron fraction, {beta}{sub eff}, one of the most important parameter in reactor kinetics, was measured for the 100 kW IPR-R1 TRIGA Mark I research reactor, located at the Nuclear Technology Development Center - CDTN, Belo Horizonte, Brazil. The current reactor core has 63 fuel elements, containing about 8.5% and 8% by weight of uranium enriched to 20% in U{sup 235}. The core has cylindrical configuration with an annular graphite reflector. Since the first criticality of the reactor in November 1960, the core configuration and the number of fuel elements have been changed several times. At that time, the reactor power was 30 kW, there were 56 fuel elements in the core, and the {beta}{sub eff} value for the reactor recommended by General Atomic (manufacturer of TRIGA) was 790 pcm. The current {beta}{sub eff} parameter was determined from experimental methods based on inhour equation and on the control rod drops. The estimated values obtained were (774 {+-} 38) pcm and (744 {+-} 20) pcm, respectively. The {beta}{sub eff} was calculated by Monte Carlo transport code MCNP5 and it was obtained 747 pcm. The calculated and measured values are in good agreement, and the relative percentage error is -3.6% for the first case, and 0.4% for the second one. (author)

  20. Genome of Herbaspirillum seropedicae strain SmR1, a specialized diazotrophic endophyte of tropical grasses.

    Fábio O Pedrosa


    Full Text Available The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme--GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species.

  1. Imaging polarimetry and spectropolarimetry of comet C/2013 R1 (Lovejoy)

    Borisov, Galin; Bagnulo, Stefano; Nikolov, Plamen; Bonev, Tanyu


    We have obtained imaging polarimetry of the comet C/2013 R1 (Lovejoy) with 2-Channel-Focal-Reducer Rozhen instrument at 2m Ritchey-Chrétien-Coudé telescope of the Bulgarian National Astronomical Observatory Rozhen in two dust continuum filters covering wavelength intervals clear from molecular emissions and centred at 4430 Å in blue filter and at 6840 Å in red filter. In imaging mode we measured the degree of linear polarisation 17.01±0.09% in the blue and 18.81±0.02% in the red, which is in a very good agreement with measurements of other comets at the similar phase angle. We have also obtained polarisation maps in both filters. We found a strong correlation between the spatial distribution of the polarisation and the dust colour. Spectropolarimetry of the nucleus region shows an increase of the polarisation with wavelength, and a depolarisation in the spectral regions with gas emission lines, most noticeable in C2 emission band, which shows a polarisation of 6.0±1.1%.

  2. Imaging polarimetry and spectropolarimetry of comet C/2013 R1 (Lovejoy)

    Borisov, Galin; Nikolov, Plamen; Bonev, Tanyu


    We have obtained imaging polarimetry of the comet C/2013 R1 (Lovejoy) with 2-Channel-Focal-Reducer Rozhen instrument at 2m Ritchey-Chr\\'etien-Coud\\'e telescope of the Bulgarian National Astronomical Observatory Rozhen in two dust continuum filters covering wavelength intervals clear from molecular emissions and centred at $4430\\AA$ in (blue filter) and at $6840\\AA$ in (red filter). In imaging mode we measured the degree of linear polarisation $17.01\\pm0.09$% in the blue and $18.81\\pm0.02$% in the red, which is in a very good agreement with measurements of other comets at the similar phase angle. We have also obtained polarisation maps in both filters. We found a strong correlation between the spatial distribution of the polarisation and the dust colour. Spectropolarimetry of the nucleus region shows an increase of the polarisation with wavelength, and a depolarisation in the spectral regions with gas emission lines, most noticeable in C$_2$ emission band, which shows a polarisation of $6.0\\pm1.1$%.

  3. Atlas of products for wave-Sobolev spaces on $\\mathbf R^{1+3}$

    D'Ancona, Piero; Selberg, Sigmund


    The wave-Sobolev spaces $H^{s,b}$ are $L^2$-based Sobolev spaces on the Minkowski space-time $\\R^{1+n}$, with Fourier weights are adapted to the symbol of the d'Alembertian. They are a standard tool in the study of regularity properties of nonlinear wave equations, and in such applications the need arises for product estimates in these spaces. Unfortunately, it seems that with every new application some estimates come up which have not yet appeared in the literature, and then one has to resort to a set of well-established procedures for proving the missing estimates. To relieve the tedium of having to constantly fill in such gaps "by hand", we make here a systematic effort to determine the complete set of estimates in the bilinear case. We determine a set of necessary conditions for a product estimate $H^{s_1,b_1} \\cdot H^{s_2,b_2} \\hookrightarrow H^{-s_0,-b_0}$ to hold. These conditions define a polyhedron $\\Omega$ in the space $\\R^6$ of exponents $(s_0,s_1,s_2,b_0,b_1,b_2)$. We then show, in space dimension...

  4. Experience on wet storage spent fuel sipping at IEA-R1 Brazilian research reactor

    Perrotta, J.A.; Terremoto, L.A.A.; Zeituni, C.A. [Instituto de Pesquisas Energeticas e Nucleares, Sao Paulo (Brazil). Divisao de Engenharia do Nucleo


    The IEA-R1 research reactor of the Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP) is a pool type reactor of B and W design, that has been operating since 1957 at a power of 2 MW. Irradiated (spent) fuels have been stored at the facility during the various years of operation. At present there are 40 spent fuel assemblies at dry storage, 79 spent fuel assemblies at wet storage and 30 fuel assemblies in the core. The oldest fuels are of United States origin, made with U-Al alloy, both of LEU and HEU MTR fuel type. many of these fuel assemblies have corrosion pits along their lateral fuel plates. These pits originate by galvanic corrosion between the fuel plate and the stainless steel storage racks. As a consequence of the possibility of sending the irradiated old fuels back to the U.S.A., sipping tests were performed with the spent fuel assemblies. The reason for this was to evaluate their {sup 137}Cs leaking rate, if any. This work describes the procedure and methodology used to perform the sipping tests with the fuel assemblies at the storage pool, and presents the results obtained for the {sup 137}Cs sipping water activity for each fuel assembly. A correlation is made between the corrosion pits and the activity values measured. A {sup 137}Cs leaking rate is determined and compared to the criteria established for canning spent fuel assemblies before shipment. (author).

  5. Experience on wet storage spent fuel sipping at IEA-R1 Brazilian research reactor

    Perrotta, J.A.; Terremoto, L.A.A.; Zeituni, C.A


    The IEA-R1 research reactor of the Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP) is a pool type reactor of B and W design, that has been operating since 1957 at a power of 2 MW. Irradiated (spent) fuels have been stored at the facility during the various years of operation. At present there are 40 spent fuel assemblies at dry storage, 79 spent fuel assemblies at wet storage and 30 fuel assemblies in the core. The oldest fuels are of United States origin, made with U-Al alloy, both of LEU and HEU MTR fuel type. Many of these fuel assemblies have corrosion pits along their lateral fuel plates. These pits originate by galvanic corrosion between the fuel plate and the stainless steel storage racks. As a consequence of the possibility of sending the irradiated old fuels back the U.S.A., sipping tests were performed with the spent fuel assemblies. The reason for this was to evaluate their {sup 137}Cs leaking rate, if any. This work describes the procedure and methodology used to perform the sipping tests with the fuel assemblies at the storage pool, and presents the results obtained for the {sup 137}Cs sipping water activity for each fuel assembly. A correlation is made between the corrosion pits and the activity values measured. A {sup 137}Cs leaking rate is determined and compared to the criteria established for canning spent fuel assemblies before shipment.

  6. Bandlike electrical transport in P r1 -xC axMn O3 manganites

    Kadyrov, L. S.; Zhang, T.; Zhukova, E. S.; Anzin, V. B.; Trotsenko, V. G.; Torgashev, V. I.; Dressel, M.; Gorshunov, B. P.


    The conductivity and dielectric permittivity spectra of polycrystalline manganites P r1 -xC axMn O3 with x =0.3 , 0.4, and 0.5 have been measured in a broad frequency range (5 -3000 c m-1 ) down to low temperatures. From the dispersion analysis the existence of a Drude component can be established that is associated with small polarons coherently moving within a band several meV wide. The temperature-dependent conductivity shows an activated behavior with an activation energy of approximately 165 meV above 170 K and 4.8 meV below 120 K. The change in the activation energy is assigned to the onset of magnetic order at 120 K. In all three samples an absorption band is discovered at 40 -60 c m-1 that is associated with the transition between Stark-split P r3 + electron states which gain optical activity due to coupling to acoustical phonons.

  7. Cloning and Function Analysis of Transcription Factor Gene NtGRAS-R1 from Nicotiana tabacum L.%烟草转录因子基因NtGRAS-R1的克隆与功能分析

    李富欣; 许芳芳; 李素敏; 肖万福; 孙艳敏; 刘卫群; 郭红祥


    为了分析NtGRAS-R1的生物学功能,在NCBI上BLAST植物的EST数据库拼接获得Nt-GRAS-R1的全长序列;根据该序列设计特异引物,利用PCR方法从烟草根系cDNA 中扩增Nt-GRAS-R1,将其连接到pS1300表达载体上,采用农杆菌介导的花序侵染法转化拟南芥,采用RT-PCR法检测转基因拟南芥植株;获得稳定转基因拟南芥后观察生长性状,并用qPCR方法检测At-CLV3基因的表达情况。结果显示,NtGRAS-R1基因属于HAM亚家族,编码508个氨基酸;观察发现,转基因拟南芥植株根长和根体积明显大于野生型;qPCR结果表明,转基因拟南芥AtCLV3的表达量明显低于野生型拟南芥。初步表明NtGRAS-R1参与根系生长发育调控过程。%The aim of this research is to explore the function of NtGRAS-R1 . The full-length sequence of NtGRAS-R1 was acquired with in silico cloning method. The ORF fragment of NtGRAS-R1 was obtained with PCR amplification method, and then pS1300-NtGRAS-R1 expression vector was constructed. Transgenic Arabidopsis with NtGRAS-R1 was obtained with Agrobacterium tumefaciens-mediated floral dip transformation method. RT-PCR was used to identify transgenic plants. The phenotypic of transgenic Arabidopsis was observed, and the expression of AtCLV3 was detected with qPCR method. The results showed that NtGRAS-R1 belonged to HAM subfamily,having an ORF of 1 527 bp to encode a protein of 508 amino acids. Compared to wild type plants, the root length and volume of transgenic Arabidopsis obviously increased. The expression of AtCLV3 in transgenic Arabidopsis was lower than wild type plant. In the present study, NtGRAS-R1 was successfully cloned, and the phenotypic analysis of transgenic Arabidopsis shows that NtGRAS-R1 has a role in the process of tobacco roots growth and development.

  8. Longitudinal T1 relaxation rate (R1) captures changes in short-term Mn exposure in welders.

    Lewis, Mechelle M; Flynn, Michael R; Lee, Eun-Young; Van Buren, Scott; Van Buren, Eric; Du, Guangwei; Fry, Rebecca C; Herring, Amy H; Kong, Lan; Mailman, Richard B; Huang, Xuemei


    We demonstrated recently that the T1 relaxation rate (R1) captured short-term Mn exposure in welders with chronic, relatively low exposure levels in a cross-sectional study. In the current study, we used a longitudinal design to examine whether R1 values reflect the short-term dynamics of Mn exposure. Twenty-nine welders were evaluated at baseline and 12 months. Occupational questionnaires estimated short-term welding exposure using welding hours in the 90days prior to each study visit (HrsW90). In addition, blood Mn levels, the pallidal index (PI; globus pallidus T1-weighted intensity (T1WI)/frontal white matter T1WI), and R1 values in brain regions of interest (ROIs) were determined as Mn biomarkers at each visit. Associations between changes in estimated welding exposure and changes in purported Mn biomarkers were assessed by Spearman's correlations with adjustment for age and baseline R1, HrsW90, and blood Mn values. Changes in welding hours (HrsW90: the short-term welding exposure estimate), was associated significantly with changes in R1 values in the putamen (r=0.541, p=0.005), caudate (R=0.453, p=0.023), globus pallidus (R=0.430, p=0.032), amygdala (R=0.461, p=0.020), and hippocampus (R=0.447, p=0.025), but not with changes in blood Mn levels or the PI. Changes in R1 values correlated with changes in the short-term welding exposure estimate, but not with more traditional measures of Mn exposure (blood Mn levels or PI). These results suggest that R1 may serve as a useful marker to capture the short-term dynamics in Mn brain accumulation related to welding exposure. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Spent fuel management plans for the FiR 1 Reactor

    Salmenhaara, S. E. J. [V1T Processes Technical Research Centre of Finland (VTT), Otakaari 3 A, P.O. Box 1404, FIN-02044 VTT, (Finland)


    The FiR 1-reactor, a 250 kW TRIGA reactor, has been in operation since 1962. The main purpose to run the reactor is now the Boron Neutron Capture Therapy (BNCT). The BNCT work dominates the current utilization of the reactor: three days per week for BNCT purposes and only two days per week for other purposes such as the neutron activation analysis and isotope production. The final disposal site is situated in Olkiluoto, on the western coast of Finland. Olkiluoto is also one of the two nuclear power plant sites in Finland. In the new operating license of our reactor there is a special condition. We have to achieve a binding agreement between our Research Centre and either the domestic Nuclear Power Companies about the possibility to use the Olkiluoto final disposal facility for our spent fuel or US DOE about the return of our spent fuel back to USA. If we want to continue the reactor operation beyond the year 2006. the domestic final disposal is the only possibility. At the moment it seems to be reasonable to prepare to both possibilities: the domestic final disposal and the return to the USA offered by US DOE. Because the cost estimates of the both possibilities are on the same order of magnitude, the future of the reactor itself will decide, which of the spent fuel policies will be obeyed. In a couple of years' time it will be seen, if the funding of the reactor and the incomes from the BNCT treatments will cover the costs. If the BNCT and other irradiations develop satisfactorily, the reactor can be kept in operation beyond the year 2006 and the domestic final disposal will be implemented. If, however, there is still lack of money, there is no reason to continue the operation of the reactor and the choice of US DOE alternative is natural. (author)

  10. A Genetic Screen To Assess Dopamine Receptor (DopR1 Dependent Sleep Regulation in Drosophila

    Yiqin Jiang


    Full Text Available Sleep is an essential behavioral state of rest that is regulated by homeostatic drives to ensure a balance of sleep and activity, as well as independent arousal mechanisms in the central brain. Dopamine has been identified as a critical regulator of both sleep behavior and arousal. Here, we present results of a genetic screen that selectively restored the Dopamine Receptor (DopR/DopR1/dumb to specific neuroanatomical regions of the adult Drosophila brain to assess requirements for DopR in sleep behavior. We have identified subsets of the mushroom body that utilizes DopR in daytime sleep regulation. These data are supported by multiple examples of spatially restricted genetic rescue data in discrete circuits of the mushroom body, as well as immunohistochemistry that corroborates the localization of DopR protein within mushroom body circuits. Independent loss of function data using an inducible RNAi construct in the same specific circuits also supports a requirement for DopR in daytime sleep. Additional circuit activation of discrete DopR+ mushroom body neurons also suggests roles for these subpopulations in sleep behavior. These conclusions support a new separable function for DopR in daytime sleep regulation within the mushroom body. This daytime regulation is independent of the known role of DopR in nighttime sleep, which is regulated within the Fan-Shaped Body (FSB. This study provides new neuroanatomical loci for exploration of dopaminergic sleep functions in Drosophila, and expands our understanding of sleep regulation during the day vs. night.

  11. Clinicopathologic findings in (anti-FcepsilonR1alpha) autoimmune-related chronic urticaria.

    Rojanapremsuk, Theera; Kasprowicz, Sarah; Schafer, Ewa; Story, Rachel; Clarke, Michael S; Walls, Timothy; Snyder, Vivian; Gleason, Briana C; Thomas, Antoinette B; Cibull, Thomas


    One cause of chronic urticaria is autoreactivity which is diagnosed by detecting autoantibodies against the IgE receptor alpha subunit (anti-Fc R1alpha). To compare the histopathologic features of chronic urticaria patients testing positive for anti-IgE receptor antibody (Ab) to those testing negative. Totally, 438 patients with a clinical presentation of chronic urticaria (2011-2013) had anti-IgE receptor Ab tested and 37 of those patients had skin biopsy. We evaluated microscopic features including: spongiosis, dermal edema, presence of mast cells, density of lymphocytic infiltration, predomination of eosinophils/neutrophils; intravascular neutrophils and presence of vasculitis. The aforementioned features were compared between negative and positive anti-IgE receptor Ab groups. Of 37 patients , 69% were women and 31% were men. 49% had positive anti-IgE receptor Ab and 51% had negative anti-IgE receptor Ab. In the positive anti-IgE receptor Ab group, 83% showed intravascular neutrophils. Eosinophil predominance was identified in 72% and neutrophil predominance was identified in 28%. In the negative anti-IgE receptor Ab group, 89% showed intravascular neutrophils. Eosinophil predominance was identified in 53% and neutrophil predominance was identified in 47%. There was no evidence of vasculitis in either group. There were no significant histopathologic differences between the anti-IgE receptor Ab positive and negative cases. Therefore, serum testing for anti-IgE receptor Ab is required to identify this subgroup of chronic urticaria patients. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  12. Characterization of filter cartridges from the IEA-R1 reactor by radiochemical method

    Geraldo, Bianca; Vicente, Roberto; Ferreira, Robson J.; Goes, Marcos M.; Marumo, Julio T., E-mail: [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    The filter cartridges used in water purification system of research nuclear reactor IEA-R1 are considered radioactive wastes after their useful life. The characterization of these wastes is one of the stages of management, which aims to identify and quantify the radionuclides present, including those known as 'difficult to measure' (DTM) radionuclides. Establish a radiochemical analysis methodology for this type of waste is a difficult job, not only by the application of these techniques, but also by the amount of radionuclides that should be analyzed. In the waste produced in a nuclear reactor, the most important radionuclides are fission products, activation products and transuranic elements. Since these radionuclides emit gamma radiation not measurable in its decay process and consequently are difficult to measure, their concentrations can be estimated by indirect methods such as scale factors. This method is used to evaluate the DTM concentration, which is represented by alpha and beta nuclides using the correlation between them and the radionuclide key, a gamma emitter. The objective of this work is to describe a radiochemical analysis methodology for gamma emitter nuclides, present in the filter cartridges, evaluating the activity and concentrations by destructive assays. At the same time, two studies have been performed by non-destructive assays, the first one based on dose rates and the point kernel method to correlate the results and the second one based on calibration efficiency with Monte Carlo method. These studies belong to the radioactive waste characterization program that has been conducted at the Waste Management Laboratory of Nuclear and Energy Research Institute, IPEN-CNEN/SP. (author)

  13. Sinorhizobium meliloti CpdR1 is critical for co-ordinating cell cycle progression and the symbiotic chronic infection.

    Kobayashi, Hajime; De Nisco, Nicole J; Chien, Peter; Simmons, Lyle A; Walker, Graham C


    ATP-driven proteolysis plays a major role in regulating the bacterial cell cycle, development and stress responses. In the nitro -fixing symbiosis with host plants, Sinorhizobium meliloti undergoes a profound cellular differentiation, including endoreduplication of the ome. The regulatory mechanisms governing the alterations of the S. meliloti cell cycle in planta are largely unknown. Here, we report the characterization of two cpdR homologues, cpdR1 and cpdR2, of S. meliloti that encode single-domain response regulators. In Caulobacter crescentus, CpdR controls the polar localization of the ClpXP protease, thereby mediating the regulated proteolysis of key protein(s), such as CtrA, involved in cell cycle progression. The S. meliloti cpdR1-null mutant can invade the host cytoplasm, however, the intracellular bacteria are unable to differentiate into bacteroids. We show that S. meliloti CpdR1 has a polar localization pattern and a role in ClpX positioning similar to C. crescentus CpdR, suggesting a conserved function of CpdR proteins among alpha-proteobacteria. However, in S. meliloti, free-living cells of the cpdR1-null mutant show a striking morphology of irregular coccoids and aberrant DNA replication. Thus, we demonstrate that CpdR1 mediates the co-ordination of cell cycle events, which are critical for both the free-living cell division and the differentiation required for the chronic intracellular infection.

  14. Inhibition of IL-1R1/MyD88 signalling promotes mesenchymal stem cell-driven tissue regeneration.

    Martino, Mikaël M; Maruyama, Kenta; Kuhn, Gisela A; Satoh, Takashi; Takeuchi, Osamu; Müller, Ralph; Akira, Shizuo


    Tissue injury and the healing response lead to the release of endogenous danger signals including Toll-like receptor (TLR) and interleukin-1 receptor, type 1 (IL-1R1) ligands, which modulate the immune microenvironment. Because TLRs and IL-1R1 have been shown to influence the repair process of various tissues, we explored their role during bone regeneration, seeking to design regenerative strategies integrating a control of their signalling. Here we show that IL-1R1/MyD88 signalling negatively regulates bone regeneration, in the mouse. Furthermore, IL-1β which is released at the bone injury site, inhibits the regenerative capacities of mesenchymal stem cells (MSCs). Mechanistically, IL-1R1/MyD88 signalling impairs MSC proliferation, migration and differentiation by inhibiting the Akt/GSK-3β/β-catenin pathway. Lastly, as a proof of concept, we engineer a MSC delivery system integrating inhibitors of IL-1R1/MyD88 signalling. Using this strategy, we considerably improve MSC-based bone regeneration in the mouse, demonstrating that this approach may be useful in regenerative medicine applications.

  15. The Effects of HZE Particles, γ and X-ray Radiation on the Survival and Genetic Integrity of Halobacterium salinarum NRC-1, Halococcus hamelinensis, and Halococcus morrhuae

    Leuko, Stefan; Rettberg, Petra


    Three halophilic archaea, Halobacterium salinarum NRC-1, Halococcus hamelinensis, and Halococcus morrhuae, have been exposed to different regimes of simulated outer space ionizing radiation. Strains were exposed to high-energy heavy ion (HZE) particles, namely iron and argon ions, as well as to γ radiation (60Co) and X-rays, and the survival and the genetic integrity of the 16S rRNA gene were evaluated. Exposure to 1 kGy of argon or iron ions at the Heavy Ion Medical Accelerator in Chiba (HIMAC) facility at the National Institute for Radiological Sciences (NIRS) in Japan did not lead to a detectable loss in viability; only after exposure to 2 kGy of iron ions a decline in survival was observed. Furthermore, a delay in growth was manifested following exposure to 2 kGy iron ions. DNA integrity of the 16S rRNA was not compromised up to 1 kGy, with the exception of Hcc. hamelinensis following exposure to argon particles. All three strains showed a high resistance toward X-rays (exposed at the DLR in Cologne, Germany), where Hcc. hamelinensis and Hcc. morrhuae displayed better survival compared to Hbt. salinarum NRC-1. In all three organisms the DNA damage increased in a dose-dependent manner. To determine a biological endpoint for survival following exposure to γ radiation, strains were exposed to up to 112 kGy at the Beta-Gamma-Service GmbH (BGS) in Germany. Although all strains were incubated for up to 4 months, only Hcc. hamelinensis and Hcc. morrhuae recovered from 6 kGy of γ radiation. In comparison, Hbt. salinarum NRC-1 did not recover. The 16S rRNA gene integrity stayed remarkably well preserved up to 48 kGy for both halococci. This research presents novel data on the survival and genetic stability of three halophilic archaea following exposure to simulated outer space radiation.

  16. MutS and MutL are dispensable for maintenance of the genomic mutation rate in the halophilic archaeon Halobacterium salinarum NRC-1.

    Courtney R Busch

    Full Text Available BACKGROUND: The genome of the halophilic archaeon Halobacterium salinarum NRC-1 encodes for homologs of MutS and MutL, which are key proteins of a DNA mismatch repair pathway conserved in Bacteria and Eukarya. Mismatch repair is essential for retaining the fidelity of genetic information and defects in this pathway result in the deleterious accumulation of mutations and in hereditary diseases in humans. METHODOLOGY/PRINCIPAL FINDINGS: We calculated the spontaneous genomic mutation rate of H. salinarum NRC-1 using fluctuation tests targeting genes of the uracil monophosphate biosynthesis pathway. We found that H. salinarum NRC-1 has a low incidence of mutation suggesting the presence of active mechanisms to control spontaneous mutations during replication. The spectrum of mutational changes found in H. salinarum NRC-1, and in other archaea, appears to be unique to this domain of life and might be a consequence of their adaption to extreme environmental conditions. In-frame targeted gene deletions of H. salinarum NRC-1 mismatch repair genes and phenotypic characterization of the mutants demonstrated that the mutS and mutL genes are not required for maintenance of the observed mutation rate. CONCLUSIONS/SIGNIFICANCE: We established that H. salinarum NRC-1 mutS and mutL genes are redundant to an alternative system that limits spontaneous mutation in this organism. This finding leads to the puzzling question of what mechanism is responsible for maintenance of the low genomic mutation rates observed in the Archaea, which for the most part do not have MutS and MutL homologs.

  17. Photoprotection of Skin Fibroblasts from Uitraviolet Radiation by Notoginsenoside R1%三七皂苷R1对UV辐射皮肤成纤维细胞的影响

    谢璟; 何黎; 郝萍; 万屏


    Objective To observe the photoprotection of notoginsenoside R1 on fibroblasts after UVB and UVA irradiation and to explore the relative molecular mechanism. Methods Fibroblasts were obtained from human skin. The cells were irradiated with different dosages of UV and treated with notoginsenoside R1. The damage of fibroblasts was observed with inverted phase contrast microscope. Cellular proliferation activity was detected by MTT method. The hy-droxyproline amount in the culture supernatant was measured by colorimetric method, and the level of matrix metal-loproteinase-1 (MMP-1) secreted by fibroblasts was measured with ELISA kits. Results The irradiation damage degree of the cells was dependent with the irradiated dose. The number and morphology of the cells remained normal when they were pretreated with notoginsenoside R1. The activity of proliferation of fibroblasts irradiated by UV was increased and MMP-1 protein secretion level was suppressed when the dose of notoginsenoside R1 was 5 μg· mL-1 and 20 μg· mL-1. Conclusion UVA and UVB irradiation can induce obvious damage to fibroblasts of human skin. Notoginsenoside R1 protects the damage of fibroblasts from UV irradiation.%目的 探讨三七皂苷R1对UV诱导的人皮肤成纤维细胞的保护作用及其相关分子机制.方法 采用不同剂量UV照射原代培养的人皮肤成纤维细胞,同时加入三七皂苷R1干预处理,倒置相差显微镜观察细胞损伤的形态学变化,并用MTT法检测其增殖活性,比色法检测成纤维细胞培养上清中羟脯氨酸的含量,ELISA法检测成纤维细胞MMP-1蛋白水平.结果 UVB、UVA辐射成纤维细胞后,细胞损伤程度呈剂量依赖性,加入三七皂苷R1后,体外培养成纤维细胞未出现明显的数量上和形态学改变;三七皂苷R1浓度为5,20 μg·mL-1时经UV照射的FB细胞增殖活性增加;三七皂苷R1浓度为5,20 μg· mL-1时抑制MMP-1的分泌.结论 UVB、UVA辐射对皮肤成纤维细胞具有明

  18. Cloning and characterization of XiR1, a locus responsible for dagger nematode resistance in grape.

    Hwang, Chin-Feng; Xu, Kenong; Hu, Rong; Zhou, Rita; Riaz, Summaira; Walker, M Andrew


    The dagger nematode, Xiphinema index, feeds aggressively on grape roots and in the process, vectors grapevine fanleaf virus (GFLV) leading to the severe viral disease known as fanleaf degeneration. Resistance to X. index and GFLV has been the key objective of grape rootstock breeding programs. A previous study found that resistance to X. index derived from Vitis arizonica was largely controlled by a major quantitative trait locus, XiR1 (X. index Resistance 1), located on chromosome 19. The study presented here develops high-resolution genetic and physical maps in an effort to identify the XiR1 gene(s). The mapping was carried out with 1,375 genotypes in three populations derived from D8909-15, a resistant selection from a cross of V. rupestris A. de Serres (susceptible) x V. arizonica b42-26 (resistant). Resistance to X. index was evaluated on 99 informative recombinants that were identified by screening the three populations with two markers flanking the XiR1 locus. The high-resolution genetic map of XiR1 was primarily constructed with seven DNA markers developed in this study. Physical mapping of XiR1 was accomplished by screening three bacterial artificial chromosome (BAC) libraries constructed from D8909-15, V. vinifera Cabernet Sauvignon and V. arizonica b42-26. A total of 32 BAC clones were identified and the XiR1 locus was delineated within a 115 kb region. Sequence analysis of three BAC clones identified putative nucleotide binding/leucine-rich repeat (NB-LRR) genes. This is the first report of a closely linked major gene locus responsible for ectoparasitic nematode resistance. The markers developed from this study are being used to expedite the breeding of resistant grape rootstocks.

  19. MicroRNA miR-1 is up-regulated in remote myocardium in patients with myocardial infarction.

    Bostjancic, E; Zidar, N; Stajner, D; Glavac, D


    MicroRNAs are small regulatory RNA molecules that mediate regulation of gene expression, thus affecting a variety of physiological, developmental and pathological conditions. They are believed to be new promising therapeutic targets. In recent studies two muscle-specific microRNAs were discovered to contribute to heart diseases and development: miR-1 and miR-133, but there is little data on their expression patterns in human myocardial infarction. We performed simultaneous expression analysis of miR-1, miR-133a, miR-133b in samples of infarcted tissue and remote myocardium from twenty- four patients with acute myocardial infarction. MicroRNA expression was analysed using quantitative real-time PCR and compared to the expression patterns in myocardium of eight healthy adults who died in accidents. We found ~3.8-fold miR-1 up-regulation in remote myocardium when compared to infarcted tissue or healthy adult hearts. As miR-1 has been shown in animal models and clinical studies to contribute to arrhythmogenesis by regulating pacemaker channel genes, our finding of miR-1 up-regulation in patients with myocardial infarction indicates that it might be responsible for the higher risk for arrhythmias in these patients. In addition, miR-133a/b down-regulation in infarcted tissue and remote myocardium was observed, indicating miR-133a/b involvement in the heart response to myocardial infarction. We conclude that miR-1 and miR-133 seem to be important regulators of heart adaptation after ischaemic stress.

  20. Chloroform extract of hog barn dust modulates skeletal muscle ryanodine receptor calcium-release channel (RyR1)

    Tian, Chengju; Shao, Chun Hong; Fenster, Danielle S.; Mixan, Mark; Romberger, Debra J.; Toews, Myron L.


    Skeletal muscle weakness is a reported ailment in individuals working in commercial hog confinement facilities. To date, specific mechanisms responsible for this symptom remain undefined. The purpose of this study was to assess whether hog barn dust (HBD) contains components that are capable of binding to and modulating the activity of type 1 ryanodine receptor Ca2+-release channel (RyR1), a key regulator of skeletal muscle function. HBD collected from confinement facilities in Nebraska were extracted with chloroform, filtered, and rotary evaporated to dryness. Residues were resuspended in hexane-chloroform (20:1) and precipitates, referred to as HBDorg, were air-dried and studied further. In competition assays, HBDorg dose-dependently displaced [3H]ryanodine from binding sites on RyR1 with an IC50 of 1.5 ± 0.1 μg/ml (Ki = 0.4 ± 0.0 μg/ml). In single-channel assays using RyR1 reconstituted into a lipid bilayer, HBDorg exhibited three distinct dose-dependent effects: first it increased the open probability of RyR1 by increasing its gating frequency and dwell time in the open state, then it induced a state of reduced conductance (55% of maximum) that was more likely to occur and persist at positive holding potentials, and finally it irreversibly closed RyR1. In differentiated C2C12 myotubes, addition of HBD triggered a rise in intracellular Ca2+ that was blocked by pretreatment with ryanodine. Since persistent activation and/or closure of RyR1 results in skeletal muscle weakness, these new data suggest that HBD is responsible, at least in part, for the muscle ailment reported by hog confinement workers. PMID:20576841

  1. Enhanced natural killer cell activation by exopolysaccharides derived from yogurt fermented with Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1.

    Makino, Seiya; Sato, Asako; Goto, Ayako; Nakamura, Marie; Ogawa, Miho; Chiba, Yoshika; Hemmi, Jun; Kano, Hiroshi; Takeda, Kazuyoshi; Okumura, Ko; Asami, Yukio


    Yogurt is generally recognized as a beneficial food for our health, but research into its physiological effects has focused mainly on intestinal dysfunctions such as constipation and diarrhea. We previously found yogurt fermented with Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 (hereafter OLL1073R-1) could reduce risks of catching the common cold and flu in human trials. It was assumed that immunostimulatory exopolysaccharide (EPS) produced from OLL1073R-1 play an important role in this context. However, few studies have examined the immunostimulatory effects of traditional Bulgarian yogurts fermented with different strains of lactobacilli and their metabolites. Therefore, we screened 139 L. delbrueckii ssp. bulgaricus strains and identified OLL1073R-1 as the most robust producer of EPS. This strain was also the only strain that induced the production of IFN-γ in vitro. Oral administration of the EPS or yogurt fermented with OLL1073R-1 and Streptococcus thermophilus OLS3059 (OLL1073R-1 yogurt) augmented natural killer (NK) cell activity and induced IFN-γ production in spleen cells in mice, whereas 2 other yogurts fermented with other strains had no effect on NK cell activity. Cellular preparations of the OLL1073R-1 strain also slightly augmented NK cell activity, but were less effective than EPS itself. The EPS-dependent stimulation of NK cell activity was abrogated in IFN-γ knockout mice and in myeloid differentiation factor 88 knockout mice. Furthermore, IFN-γ production from spleen cells stimulated with EPS was completely blocked with both anti-IL-12 and anti-IL-18 antibodies in vitro. These findings suggest that NK cell activation by OLL1073R-1 yogurt is EPS-dependent, occurs via IL-12- and IL-18-mediated IFN-γ production, and requires myeloid differentiation factor 88. We showed that traditional Bulgarian yogurt could exert immunostimulatory effects by selecting starter strains and part of the mechanisms depend on IFN-γ inducible EPS produced

  2. Developing new PET tracers to image the growth hormone secretagogue receptor 1a (GHS-R1a).

    Kawamura, Kazunori; Fujinaga, Masayuki; Shimoda, Yoko; Yamasaki, Tomoteru; Zhang, Yiding; Hatori, Akiko; Xie, Lin; Wakizaka, Hidekatsu; Kumata, Katsushi; Ohkubo, Takayuki; Kurihara, Yusuke; Ogawa, Masanao; Nengaki, Nobuki; Zhang, Ming-Rong


    `The growth hormone secretagogue receptor 1a (GHS-R1a) is the orphan G-protein-coupled receptor, and its endogenous ligand is ghrelin. GHS-R1a contributes to regulation of glucose homeostasis, memory and learning, food addiction, and neuroprotection. Several PET tracers for GHS-R1a have been developed, but none have been reported to be clinically applicable to GHS-R1a imaging. In this study, we developed three new PET tracers for GHS-R1a: (18)F-labeled 6-(4-chlorophenyl)-3-((1-(2-fluoroethyl)piperidin-3-yl)methyl)-2-(o-tolyl)quinazolin-4(3H)-one (1), (11)C-labeled 6-(4-chlorophenyl)-3-((1-(2-methoxyethyl)piperidin-3-yl)methyl)-2-(o-tolyl)quinazolin-4(3H)-one (2), and (11)C-labeled (S)-(4-(1H-indole-6-carbonyl)-3-methylpiperazin-1-yl)(4'-methoxy-[1,1'-biphenyl]-4-yl)methanone (3). [(18)F]1 was synthesized by the (18)F-fluoroethylation; [(11)C]2 or [(11)C]3 was synthesized by the (11)C-methylation. Biodistribution studies and PET studies were conducted in mice. We successfully radiosynthesized [(18)F]1, [(11)C]2, and [(11)C]3 with appropriate radioactivity for the animal study. In the ex vivo biodistribution study, 60min following injection, the radioactivity level of [(18)F]1 was relatively high in the small intestine, that of [(11)C]2 was high in the liver, and that of [(11)C]3 was high in the pancreas. The radioactivity levels of the three PET tracers were relatively low in the brain. Under pretreatment with YIL781 (a selective and high affinity antagonist for GHS-R1a), the pancreas radioactivity level at 30min following [(11)C]3 injection was significantly reduced to 55% of control, but the radioactivity in the brain was not changed. In the PET study under control conditions, high radioactivity levels in the liver and pancreas were observed following [(11)C]3 injection. With YIL781 pretreatment, the accumulated radioactivity in the pancreas 15-60min after [(11)C]3 injection was significantly decreased to 78% of control. [(11)C]3 exhibited relatively high uptake

  3. The internal Cdc20 binding site in BubR1 facilitates both spindle assembly checkpoint signalling and silencing

    Lischetti, Tiziana; Zhang, Gang; Sedgwick, Garry G;


    Improperly attached kinetochores activate the spindle assembly checkpoint (SAC) and by an unknown mechanism catalyse the binding of two checkpoint proteins, Mad2 and BubR1, to Cdc20 forming the mitotic checkpoint complex (MCC). Here, to address the functional role of Cdc20 kinetochore localization...... on the SAC because the IC20BD is also required for efficient SAC silencing. Indeed, the IC20BD can disrupt the MCC providing a mechanism for its role in SAC silencing. We thus uncover an unexpected dual function of the second Cdc20 binding site in BubR1 in promoting both efficient SAC signalling and SAC...

  4. A Saccharomyces cerevisiae assay system to investigate ligand/AdipoR1 interactions that lead to cellular signaling.

    Mustapha Aouida

    Full Text Available Adiponectin is a mammalian hormone that exerts anti-diabetic, anti-cancer and cardioprotective effects through interaction with its major ubiquitously expressed plasma membrane localized receptors, AdipoR1 and AdipoR2. Here, we report a Saccharomyces cerevisiae based method for investigating agonist-AdipoR interactions that is amenable for high-throughput scale-up and can be used to study both AdipoRs separately. Agonist-AdipoR1 interactions are detected using a split firefly luciferase assay based on reconstitution of firefly luciferase (Luc activity due to juxtaposition of its N- and C-terminal fragments, NLuc and CLuc, by ligand induced interaction of the chimeric proteins CLuc-AdipoR1 and APPL1-NLuc (adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif 1-NLuc in a S. cerevisiae strain lacking the yeast homolog of AdipoRs (Izh2p. The assay monitors the earliest known step in the adiponectin-AdipoR anti-diabetic signaling cascade. We demonstrate that reconstituted Luc activity can be detected in colonies or cells using a CCD camera and quantified in cell suspensions using a microplate reader. AdipoR1-APPL1 interaction occurs in absence of ligand but can be stimulated specifically by agonists such as adiponectin and the tobacco protein osmotin that was shown to have AdipoR-dependent adiponectin-like biological activity in mammalian cells. To further validate this assay, we have modeled the three dimensional structures of receptor-ligand complexes of membrane-embedded AdipoR1 with cyclic peptides derived from osmotin or osmotin-like plant proteins. We demonstrate that the calculated AdipoR1-peptide binding energies correlate with the peptides' ability to behave as AdipoR1 agonists in the split luciferase assay. Further, we demonstrate agonist-AdipoR dependent activation of protein kinase A (PKA signaling and AMP activated protein kinase (AMPK phosphorylation in S. cerevisiae, which are

  5. A Saccharomyces cerevisiae Assay System to Investigate Ligand/AdipoR1 Interactions That Lead to Cellular Signaling

    Aouida, Mustapha


    Adiponectin is a mammalian hormone that exerts anti-diabetic, anti-cancer and cardioprotective effects through interaction with its major ubiquitously expressed plasma membrane localized receptors, AdipoR1 and AdipoR2. Here, we report a Saccharomyces cerevisiae based method for investigating agonist-AdipoR interactions that is amenable for high-throughput scale-up and can be used to study both AdipoRs separately. Agonist-AdipoR1 interactions are detected using a split firefly luciferase assay based on reconstitution of firefly luciferase (Luc) activity due to juxtaposition of its N- and C-terminal fragments, NLuc and CLuc, by ligand induced interaction of the chimeric proteins CLuc-AdipoR1 and APPL1-NLuc (adaptor protein containing pleckstrin homology domain, phosphotyrosine binding domain and leucine zipper motif 1-NLuc) in a S. cerevisiae strain lacking the yeast homolog of AdipoRs (Izh2p). The assay monitors the earliest known step in the adiponectin-AdipoR anti-diabetic signaling cascade. We demonstrate that reconstituted Luc activity can be detected in colonies or cells using a CCD camera and quantified in cell suspensions using a microplate reader. AdipoR1-APPL1 interaction occurs in absence of ligand but can be stimulated specifically by agonists such as adiponectin and the tobacco protein osmotin that was shown to have AdipoR-dependent adiponectin-like biological activity in mammalian cells. To further validate this assay, we have modeled the three dimensional structures of receptor-ligand complexes of membrane-embedded AdipoR1 with cyclic peptides derived from osmotin or osmotin-like plant proteins. We demonstrate that the calculated AdipoR1-peptide binding energies correlate with the peptides\\' ability to behave as AdipoR1 agonists in the split luciferase assay. Further, we demonstrate agonist-AdipoR dependent activation of protein kinase A (PKA) signaling and AMP activated protein kinase (AMPK) phosphorylation in S. cerevisiae, which are homologous to

  6. Regulation of Fear Extinction in the Basolateral Amygdala by Dopamine D2 Receptors Accompanied by Altered GluR1, GluR1-Ser845 and NR2B Levels.

    Shi, Yan-Wei; Fan, Bu-Fang; Xue, Li; Wen, Jia-Ling; Zhao, Hu


    The amygdala, a critical structure for both Pavlovian fear conditioning and fear extinction, receives sparse but comprehensive dopamine innervation and contains dopamine D1 and D2 receptors. Fear extinction, which involves learning to suppress the expression of a previously learned fear, appears to require the dopaminergic system. The specific roles of D2 receptors in mediating associative learning underlying fear extinction require further study. Intra-basolateral amygdala (BLA) infusions of a D2 receptor agonist, quinpirole, and a D2 receptor antagonist, sulpiride, prior to fear extinction and extinction retention were tested 24 h after fear extinction training for long-term memory (LTM). LTM was facilitated by quinpirole and attenuated by sulpiride. In addition, A-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor glutamate receptor 1 (GluR1) subunit, GluR1 phospho-Ser845, and N-methyl-D-aspartic acid receptor NR2B subunit levels in the BLA were generally increased by quinpirole and down-regulated by sulpiride. The present study suggests that activation of D2 receptors facilitates fear extinction and that blockade of D2 receptors impairs fear extinction, accompanied by changes in GluR1, GluR1-Ser845 and NR2B levels in the amygdala.

  7. New digital control system for the operation of the Colombian research reactor IAN-R1; Nuevo sistema de control digital para la operacion del reactor de investigacion Colombiano IAN-R1

    Celis del A, L.; Rivero, T.; Bucio, F.; Ramirez, R.; Segovia, A.; Palacios, J., E-mail: [ININ, Carretera Mexico-Toluca s/n, 52750 Ocoyoacac, Estado de Mexico (Mexico)


    En 2011, Mexico won the Colombian international tender for the renewal of instrumentation and control of the IAN-R1 Reactor, to Argentina and the United States. This paper presents the design criteria and the development made for the new digital control system installed in the Colombian nuclear reactor IAN-R1, which is based on a redundant and diverse architecture, which provides increased availability, reliability and safety in the reactor operation. This control system and associated instrumentation met all national export requirements, with the safety requirements established by the IAEA as well as the requirements demanded by the Colombian Regulatory Body in nuclear matter. On August 20, 2012, the Colombian IAN-R1 reactor reached its first criticality controlled with the new system developed at Instituto Nacional de Investigaciones Nucleares (ININ). On September 14, 2012, the new control system of the Colombian IAN-R1 reactor was officially handed over to the Colombian authorities, this being the first time that Mexico exported nuclear technology through the ININ. Currently the reactor is operating successfully with the new control system, and has an operating license for 5 years. (Author)

  8. RSK1 protects P-glycoprotein/ABCB1 against ubiquitin–proteasomal degradation by downregulating the ubiquitin-conjugating enzyme E2 R1

    Katayama, Kazuhiro; Fujiwara, Chiaki; Noguchi, Kohji; Sugimoto, Yoshikazu


    P-glycoprotein (P-gp) is a critical determinant of multidrug resistance in cancer. We previously reported that MAPK inhibition downregulates P-gp expression and that P-gp undergoes ubiquitin–proteasomal degradation regulated by UBE2R1 and SCFFbx15. Here, we investigated the crosstalk between MAPK inhibition and the ubiquitin–proteasomal degradation of P-gp. Proteasome inhibitors or knockdown of FBXO15 and/or UBE2R1 cancelled MEK inhibitor-induced P-gp downregulation. RSK1 phosphorylated Thr162 on UBE2R1 but did not phosphorylate FBXO15. MEK and RSK inhibitors increased UBE2R1-WT but not UBE2R1-T162D and -T162A expression. UBE2R1-T162D showed higher self-ubiquitination and destabilisation than UBE2R1-WT and -T162A. Unlike UBE2R1-WT and -T162A, UBE2R1-T162D did not induce P-gp ubiquitination. UBE2R1-WT or -T162A downregulated P-gp expression and upregulated rhodamine 123 level and sensitivity to vincristine and doxorubicin. However, UBE2R1-T162D did not confer any change in P-gp expression, rhodamine 123 accumulation and sensitivity to the drugs. These results suggest that RSK1 protects P-gp against ubiquitination by reducing UBE2R1 stability. PMID:27786305

  9. The architecture of the BubR1 tetratricopeptide tandem repeat defines a protein motif underlying mitotic checkpoint-kinetochore communication

    Bolanos-Garcia, Victor M; Nilsson, Jakob; Blundell, Tom L


    as substitution of BubR1 residues engaged in KNL1 binding impaired the SAC and BubR1 recruitment into checkpoint complexes in stable cell lines. Here we discuss the implications of the disorder-to-order transition of KNL1 upon BubR1 binding for SAC signaling and propose a mechanistic model of how BUBs binding may...

  10. 利用酿酒酵母R1再生废轮胎橡胶的研究%Preliminary study of the reclaimation of ground tyre rubber by Saccharomyces cerevisiae R1

    陶子溢; 王雅琴; 赵素合; 李元虎


    The yeast species S. Cerevisiae R1 has the capability to recycle waste rubber. However the many toxic additives in rubber have a significant inhibiting effect on the growth of S. Cerevisiae R1, as shown by our toxicity tests. Ethanol leaching of the rubber material can remove some of these toxic additives, reducing the adverse impact on the growth of the microorganism. After 132 hours of regeneration, a remarkable decrease in crosslink density, increase in swelling ratio(4. 6% ) , and better physical properties were observed for the resulting styrene butadiene rubber/devulcanized ground tyre rubber (SBR/DGTR) blend, compared with the corresponding values for untreated SBR/GTR. Scanning electron microscopy images of SBR/DGTR also showed an improvement in interfacial binding force and good adhesion. X-ray photoelectron spectroscopy showed that there was no obvious change in carbon content, but the content of sulfur decreased markedly(23. 86% ) , indicative of a significant desulfurization effect.%通过毒性实验分析了废轮胎橡胶( GTR)主要组分对酿酒酵母(S.cerevisiae)R1生长的影响,并用乙醇解毒,减少其对S.cerevisiae R1的抑制作用.经132 h再生后,30份再生GTR填充到丁苯橡胶(SBR)基体中的力学性能均好于未经S.cerevisiae R1再生的硫化橡胶,其溶胀值增加4.6%,交联密度下降,撕裂强度提高了26%.再生GTR与SBR界面接触良好,XPS测试显示C元素基本不变,S元素质量分数降低了23.86%,具有显著的脱硫效果.

  11. Attenuated Pik3r1 Expression Prevents Insulin Resistance and Adipose Tissue Macrophage Accumulation in Diet-Induced Obese Mice

    McCurdy, Carrie E.; Schenk, Simon; Holliday, Michael J.; Philp, Andrew; Houck, Julie A.; Patsouris, David; MacLean, Paul S.; Majka, Susan M.; Klemm, Dwight J.; Friedman, Jacob E.


    Obese white adipose tissue (AT) is characterized by large-scale infiltration of proinflammatory macrophages, in parallel with systemic insulin resistance; however, the cellular stimulus that initiates this signaling cascade and chemokine release is still unknown. The objective of this study was to determine the role of the phosphoinositide 3-kinase (PI3K) regulatory subunits on AT macrophage (ATM) infiltration in obesity. Here, we find that the Pik3r1 regulatory subunits (i.e., p85α/p55α/p50α) are highly induced in AT from high-fat diet–fed obese mice, concurrent with insulin resistance. Global heterozygous deletion of the Pik3r1 regulatory subunits (αHZ), but not knockout of Pik3r2 (p85β), preserves whole-body, AT, and skeletal muscle insulin sensitivity, despite severe obesity. Moreover, ATM accumulation, proinflammatory gene expression, and ex vivo chemokine secretion in obese αHZ mice are markedly reduced despite endoplasmic reticulum (ER) stress, hypoxia, adipocyte hypertrophy, and Jun NH2-terminal kinase activation. Furthermore, bone marrow transplant studies reveal that these improvements in obese αHZ mice are independent of reduced Pik3r1 expression in the hematopoietic compartment. Taken together, these studies demonstrate that Pik3r1 expression plays a critical role in mediating AT insulin sensitivity and, more so, suggest that reduced PI3K activity is a key step in the initiation and propagation of the inflammatory response in obese AT. PMID:22698915

  12. Pharmacological properties of homomeric and heteromeric GluR1o and GluR3o receptors

    Nielsen, B S; Banke, T G; Schousboe, A


    Homomeric and heteromeric alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor subunits GluR1o and GluR3o were expressed in Spodoptera frugiperda (Sf9) insect cells. Membranes containing the recombinant receptors showed a doublet of bands of the expected size (99-109 kDa) after...

  13. Investigation of signal transduction routes within the sensor/transducer protein BlaR1 of Staphylococcus aureus.

    Staude, Michael W; Frederick, Thomas E; Natarajan, Sivanandam V; Wilson, Brian D; Tanner, Carol E; Ruggiero, Steven T; Mobashery, Shahriar; Peng, Jeffrey W


    The transmembrane antibiotic sensor/signal transducer protein BlaR1 is part of a cohort of proteins that confer β-lactam antibiotic resistance in methicillin-resistant Staphylococcus aureus (MRSA) [Fisher, J. F., Meroueh, S. O., and Mobashery, S. (2005) Chem. Rev. 105, 395-424; Llarrull, L. I., Fisher, J. F., and Mobashery, S. (2009) Antimicrob. Agents Chemother. 53, 4051-4063; Llarrull, L. I., Toth, M., Champion, M. M., and Mobashery, S. (2011) J. Biol. Chem. 286, 38148-38158]. Specifically, BlaR1 regulates the inducible expression of β-lactamases that hydrolytically destroy β-lactam antibiotics. The resistance phenotype starts with β-lactam antibiotic acylation of the BlaR1 extracellular domain (BlaRS). The acylation activates the cytoplasmic protease domain through an obscure signal transduction mechanism. Here, we compare protein dynamics of apo versus antibiotic-acylated BlaRS using nuclear magnetic resonance. Our analyses reveal inter-residue interactions that relay acylation-induced perturbations within the antibiotic-binding site to the transmembrane helix regions near the membrane surface. These are the first insights into the process of signal transduction by BlaR1.

  14. Mouse oocytes depend on BubR1 for proper chromosome segregation but not for prophase I arrest

    Touati, S.A.; Buffin, E.; Cladiere, D.; Hached, K.; Rachez, C.; Deursen, J.M.A. van; Wassmann, K.


    Mammalian female meiosis is error prone, with rates of meiotic chromosome missegregations strongly increasing towards the end of the reproductive lifespan. A strong reduction of BubR1 has been observed in oocytes of women approaching menopause and in ovaries of aged mice, which led to the hypothesis

  15. Normas do Teste de Inteligência Não Verbal R-1 para adultos não alfabetizados

    Maria Olinda Gottsfritz


    Full Text Available The R-1: Non-Verbal Intelligence Test is a Brazilian test to measure general intelligence. It was created to assess future drivers. However, the last published test manual does not have standards for illiterate adults. This study was designed to establish norms in the R-1 Test among illiterate adults and to present an analysis of frequencies of success for each item. The sample was composed by 151 adults, aged from 16 to 77 years of both sexes, enrolled in literacy courses in São Paulo city. The R-1 Test was collectively administered, with no time limits. Results indicated that illiterate adult’s means were lower than more educated people, confirming that education influences intelligence test scores, as it was suggested by literature. A comparison of the relative frequencies of passing individual items between the illiterate group and the more educated sample, revealed that items presented the same relative difficulty to both, although success percentages were lower for the illiterate group.   Keywords: R-1 Test; illiteracy; intellectual development; intelligence tests.

  16. Efficient bioremediation of radioactive iodine using biogenic gold nanomaterial-containing radiation-resistant bacterium, Deinococcus radiodurans R1.

    Choi, Mi Hee; Jeong, Sun-Wook; Shim, Ha Eun; Yun, Seong-Jae; Mushtaq, Sajid; Choi, Dae Seong; Jang, Beom-Su; Yang, Jung Eun; Choi, Yong Jun; Jeon, Jongho


    We herein report a new bioremediation method using a radiation-resistant bacterium. Biogenic gold nanomaterial-containing Deinococcus radiodurans R1 showed excellent capability for the removal of radioactive iodine (>99%) in several aqueous solutions. These observations demonstrated that our remediation system would be efficiently applied to the treatment of radioactive wastes.

  17. Characterization of spent fuel elements stored at IEA-R1 research reactor based on visual inspections and sipping tests

    Silva, Jose Eduardo Rosa da; Terremoto, Luis Antonio Albiac; Teodoro, Celso Antonio; Castanheira, Myrthes; Lucki, Georgi; Damy, Margaret de Almeida; Silva, Antonio Teixeira e [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail:


    Aluminum spent nuclear fuels are susceptible to corrosion attack, or mechanical damage from improper handling, while in pool reactor storage. Storage practices have been modified to reduce the potential for damage, based on recommendations presented at second WS on Spent Fuel Characterization, promoted by IAEA. In this work, we present the inspection program proposed to the IEA-R1 stored spent fuel elements, in order to provide information on the physical condition during the interim storage time under wet condition at the reactor pool. The inspection program is based on non-destructive tests results (visual inspection and sipping tests) already periodically performed to exam the IEA-R1 stored spent fuel and fuel elements from the core reactor. To record the available information and examination results it was elaborated a document in the format of a catalogue containing the proposed inspection program for the IEA-R1 stored spent fuel, the description of the visual inspection and sipping tests systems, a compilation of information and images result from the tests performed for all stored standard spent fuel element and, in annexes, copies of the reference documents. That document constitutes an important step of the effective implementation of the referred IEA-R1 spent fuel inspection program and can be used to address regulatory and operational needs for the demonstration, for example, of safe storage throughout the pool storage period. (author)

  18. Mouse oocytes depend on BubR1 for proper chromosome segregation but not for prophase I arrest

    Touati, S.A.; Buffin, E.; Cladiere, D.; Hached, K.; Rachez, C.; Deursen, J.M.A. van; Wassmann, K.


    Mammalian female meiosis is error prone, with rates of meiotic chromosome missegregations strongly increasing towards the end of the reproductive lifespan. A strong reduction of BubR1 has been observed in oocytes of women approaching menopause and in ovaries of aged mice, which led to the hypothesis

  19. Eclipse period of R1 plasmids during downshift from elevated copy number: Nonrandom selection of copies for replication.

    Olsson, Jan A; Berg, Otto; Nordström, Kurt; Dasgupta, Santanu


    The classical Meselson-Stahl density-shift method was used to study replication of pOU71, a runaway-replication derivative of plasmid R1 in Escherichia coli. The miniplasmid maintained the normal low copy number of R1 during steady growth at 30°C, but as growth temperatures were raised above 34°C, the copy number of the plasmid increased to higher levels, and at 42°C, it replicated without control in a runaway replication mode with lethal consequences for the host. The eclipse periods (minimum time between successive replication of the same DNA) of the plasmid shortened with rising copy numbers at increasing growth temperatures (Olsson et al., 2003). In this work, eclipse periods were measured during downshifts in copy number of pOU71 after it had replicated at 39 and 42°C, resulting in 7- and 50-fold higher than normal plasmid copy number per cell, respectively. Eclipse periods for plasmid replication, measured during copy number downshift, suggested that plasmid R1, normally selected randomly for replication, showed a bias such that a newly replicated DNA had a higher probability of replication compared to the bulk of the R1 population. However, even the unexpected nonrandom replication followed the copy number kinetics such that every generation, the plasmids underwent the normal inherited number of replication, n, independent of the actual number of plasmid copies in a newborn cell.

  20. Pik3r1 Is Required for Glucocorticoid-Induced Perilipin 1 Phosphorylation in Lipid Droplet for Adipocyte Lipolysis.

    Kuo, Taiyi; Chen, Tzu-Chieh; Lee, Rebecca A; Nguyen, Nguyen Huynh Thao; Broughton, Augusta E; Zhang, Danyun; Wang, Jen-Chywan


    Glucocorticoids promote lipolysis in white adipose tissue (WAT) to adapt to energy demands under stress, whereas superfluous lipolysis causes metabolic disorders, including dyslipidemia and hepatic steatosis. Glucocorticoid-induced lipolysis requires the phosphorylation of cytosolic hormone-sensitive lipase (HSL) and perilipin 1 (Plin1) in the lipid droplet by protein kinase A (PKA). We previously identified Pik3r1 (also called p85α) as a glucocorticoid receptor target gene. Here, we found that glucocorticoids increased HSL phosphorylation, but not Plin1 phosphorylation, in adipose tissue-specific Pik3r1-null (AKO) mice. Furthermore, in lipid droplets, the phosphorylation of HSL and Plin1 and the levels of catalytic and regulatory subunits of PKA were increased by glucocorticoids in wild-type mice. However, these effects were attenuated in AKO mice. In agreement with reduced WAT lipolysis, glucocorticoid- initiated hepatic steatosis and hypertriglyceridemia were improved in AKO mice. Our data demonstrated a novel role of Pik3r1 that was independent of the regulatory function of phosphoinositide 3-kinase in mediating the metabolic action of glucocorticoids. Thus, the inhibition of Pik3r1 in adipocytes could alleviate lipid disorders caused by excess glucocorticoid exposure. © 2017 by the American Diabetes Association.

  1. Safety analysis of 5 MW IEAR-1 reactor; Analise de seguranca do reator IEA-R1 a 5 MW

    Silva, Antonio T. e; Maprelian, Eduardo; Rodrigues, Antonio C.I.; Cabral, Eduardo L.L.; Molnary, Leslie de; Mesquita, Ricardo N.; Mendonca, Arlindo G. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Dept. de Reatores. E-mail:


    This paper presents the methods and procedures utilized in the safety analysis of IEA-R1 research reactor. Four postulated accidents are quantitatively analyzed, being the fuel channel blockage accident considered as the Maximum Credible Accident for the reactor. The potential accident consequences and the criteria for radiological doses acceptance are evaluated and discussed. (author)

  2. Application of Nondestructive Methods for Qualification of High Density Fuels in the IEA-R1 Reactor

    Silva, J.E.R.; Silva, A.T.; Domingos, D.B.; Terremoto, L.A.A. [Instituto de Pesquisas Energeticas e Nucleares, Comissao Nacional de Energia Nuclear (IPEN-CNEN/SP), Av.Prof. Lineu Prestes 2242, Cidade Universitaria 05508-000, Sao Paulo, SP (Brazil)


    The IEA-R1 reactor of IPEN/CNEN-SP in Brazil is a pool type research reactor cooled and moderated by demineralised water and having Beryllium and Graphite as reflectors. Since 1990, IPEN/CNEN-SP has been fabricating and qualifying its own U{sub 3}O{sub 8}-Al and U{sub 3}Si{sub 2}-Al dispersion fuels. The U{sub 3}O{sub 8}-Al dispersion fuel is qualified to a uranium density of 2.3 gU/cm{sup 3} and the U{sub 3}Si{sub 2}-Al dispersion fuel up to 3.0 gU/cm{sup 3}. The IEA-R1 reactor core is constituted of the fuels above, with low enrichment in U-235 (19.9% of U-235). Nowadays, IPEN/CNEN-SP is interested in qualifying the above dispersion fuels at higher densities. Fuel miniplates of U{sub 3}O{sub 8}-Al and U{sub 3}Si{sub 2}-Al fuels, with densities of 3.0 gU/cm{sup 3} and 4.8 gU/cm{sup 3}, respectively, which are the maximal uranium densities qualified worldwide for these dispersion fuels, were fabricated at IPEN/CNEN-SP. The miniplates were put in an irradiation device, with similar external dimensions of IEA-R1 fuel assemblies, which was placed in a peripheral position of the IEA-R1 reactor core. IPEN/CNEN-SP has no hot cells to provide destructive analysis of the irradiated fuel. As a consequence, non destructive methods are being used to evaluate irradiation performance of the fuel miniplates: i) monitoring the fuel miniplate performance during the IEA-R1 operation for the following parameters: reactor power, time of operation, neutron flux at the position of each fuel assembly, burnup, inlet and outlet water, and radiochemistry analysis of reactor water; ii) periodic underwater visual inspection of fuel miniplates and eventual sipping test for the fuel miniplate suspected of leakage. The miniplates are being periodically visually inspected by an underwater radiation-resistant camera inside the IEA-R1 reactor pool, to verify its integrity and its general plate surface conditions. A new special system was designed for the fuel miniplate swelling evaluation. The

  3. Identification of potential small molecule allosteric modulator sites on IL-1R1 ectodomain using accelerated conformational sampling method.

    Chao-Yie Yang

    Full Text Available The interleukin-1 receptor (IL-1R is the founding member of the interleukin 1 receptor family which activates innate immune response by its binding to cytokines. Reports showed dysregulation of cytokine production leads to aberrant immune cells activation which contributes to auto-inflammatory disorders and diseases. Current therapeutic strategies focus on utilizing antibodies or chimeric cytokine biologics. The large protein-protein interaction interface between cytokine receptor and cytokine poses a challenge in identifying binding sites for small molecule inhibitor development. Based on the significant conformational change of IL-1R type 1 (IL-1R1 ectodomain upon binding to different ligands observed in crystal structures, we hypothesized that transient small molecule binding sites may exist when IL-1R1 undergoes conformational transition and thus suitable for inhibitor development. Here, we employed accelerated molecular dynamics (MD simulation to efficiently sample conformational space of IL-1R1 ectodomain. Representative IL-1R1 ectodomain conformations determined from the hierarchy cluster analysis were analyzed by the SiteMap program which leads to identify small molecule binding sites at the protein-protein interaction interface and allosteric modulator locations. The cosolvent mapping analysis using phenol as the probe molecule further confirms the allosteric modulator site as a binding hotspot. Eight highest ranked fragment molecules identified from in silico screening at the modulator site were evaluated by MD simulations. Four of them restricted the IL-1R1 dynamical motion to inactive conformational space. The strategy from this study, subject to in vitro experimental validation, can be useful to identify small molecule compounds targeting the allosteric modulator sites of IL-1R and prevent IL-1R from binding to cytokine by trapping IL-1R in inactive conformations.

  4. Vitamin D insufficiency in Arabs and South Asians positively associates with polymorphisms in GC and CYP2R1 genes.

    Naser Elkum

    Full Text Available A number of genetic studies have reported an association between vitamin D related genes such as group-specific component gene (GC, Cytochrome P450, family 2, subfamily R, polypeptide 1 (CYP2R1 and 7-dehydrocholesterol reductase/nicotinamide-adenine dinucleotide synthetase 1 (DHCR7/NADSYN1 and serum levels of the active form of Vitamin D, 25 (OH D among African Americans, Caucasians, and Chinese. Little is known about how genetic variations associate with, or contribute to, 25(OHD levels in Arabs populations.Allele frequencies of 18 SNPs derived from CYP2R1, GC, and DHCR7/NADSYN1 genes in 1549 individuals (Arabs, South Asians, and Southeast Asians living in Kuwait were determined using real time genotyping assays. Serum levels of 25(OHD were measured using chemiluminescence immunoassay.GC gene polymorphisms (rs17467825, rs3755967, rs2282679, rs7041 and rs2298850 were found to be associated with 25(OHD serum levels in Arabs and South Asians. Two of the CYP2R1 SNPs (rs10500804 and rs12794714 and one of GC SNPs (rs1155563 were found to be significantly associated with 25(OHD serum levels only in people of Arab origin. Across all three ethnicities none of the SNPs of DHCR7/NADSYN1 were associated with serum 25(OHD levels and none of the 18 SNPs were significantly associated with serum 25(OHD levels in people from South East Asia.Our data show for the first time significant association between the GC (rs2282679 and rs7041, CYP2R1 (rs10741657 SNPs and 25(OHD levels. This supports their roles in vitamin D Insufficiency in Arab and South Asian populations respectively. Interestingly, two of the CYP2R1 SNPs (rs10500804 and rs12794714 and one GC SNP (rs1155563 were found to correlate with vitamin D in Arab population exclusively signifying their importance in this population.

  5. Preparation of (R)-1,2-propanediol through asymmetric reduction with bakers yeast%酵母不对称催化制备R-1,2-丙二醇

    陈勇; 李凤梅; 韩荣伟; 杜德红; 于春娣


    研究了采用面包酵母还原丙酮醇制备R-1,2-丙二醇的工艺.采用摇瓶对转化条件进行单因素实验,确定最优转化条件:丙酮醇浓度0.3 mmol/mL,pH 7.0,酵母质量浓度150 g/L,乙醇浓度为0.3 mmol/mL,转化时间25 h.在此条件下,采用分批流加策略进行1.5 L规模发酵罐转化试验,转化25 h后,发酵液的R-1,2-丙二醇浓度为0.27 mmol/mL.

  6. Effect of MLP on Expression of AdipoR1 in Diabetic Nephropathy Rats%桑叶多糖对糖尿病肾病大鼠AdipoR1表达的影响

    韩智学; 薛继婷; 李齐; 宋铁军; 王桂云; 刘洪凤


    观察桑叶多糖(mulberry leaves polysaccharide,MLP)对糖尿病肾病(diaketic nephropathy,DN)大鼠空腹血糖(FBG)、血脂水平及肾脏脂联素受体1(AdipoR1)基因表达的影响,探讨桑叶多糖对糖尿病肾病大鼠肾脏保护作用及机制.

  7. Presence of Cryptosporidium scrofarum, C. suis and C. parvum subtypes IIaA16G2R1 and IIaA13G1R1 in Eurasian wild boars (Sus scrofa).

    García-Presedo, Ignacio; Pedraza-Díaz, Susana; González-Warleta, Marta; Mezo, Mercedes; Gómez-Bautista, Mercedes; Ortega-Mora, Luis Miguel; Castro-Hermida, José Antonio


    The aim of the present study was to identify the species of Cryptosporidium infecting Eurasian wild boars (Sus scrofa) in Galicia (NW, Spain). A sampling of 209 wild boars shot in different game preserves was carried out during the hunting season in 2009-2010. All samples were examined for Cryptosporidium infection, using both immunological and molecular tools. Cryptosporidium oocysts in faecal samples were identified using a direct immunofluorescence technique with monoclonal antibodies (DFA). The presence of Cryptosporidium DNA was determined using nested PCR involving amplification of a fragment of the small-subunit (SSU) ribosomal RNA gene (SSU rRNA). A total of 35 (16.7%) samples tested positive with both techniques. However, sequencing was only possible in 27 samples. Cryptosporidium scrofarum, Cryptosporidium suis and Cryptosporidium parvum oocysts were identified in 19, 5 and 3 of the samples, respectively. Moreover, C. scrofarum was detected as a dominant species infecting all age groups (juveniles, sub adults and adults). Sequence analyses of the glycoprotein (GP60) gene revealed the presence of C. parvum subtypes IIaA16G2R1 in 2 juveniles and IIaA13G1R1 in 1 sub adult wild boar. These species and subtypes have previously been described in human patients, indicating that isolates from asymptomatic wild boars might have zoonotic potential. This is the first report of the presence of C. scrofarum, C. suis and C. parvum subtypes IIaA16G2R1 and IIaA13G1R1 in wild boars (S. scrofa) in Spain.

  8. Teste de Inteligência R1-Forma B e G36: evidência de validade convergente R1-Forma B Intelligence Test and G36: evidences of convergent validity

    Acácia A. Angeli dos Santos


    Full Text Available Considerando-se a relevância de se obter medidas confiáveis para a avaliação da inteligência, este estudo foi proposto com o objetivo de verificar a validade convergente entre os testes R1-Forma B e o G36. Participaram 78 alunos que freqüentavam o Telecurso do ensino médio, com idades entre 15 e 64 anos (média = 36,84; DP = 10,10, sendo 27 (34,6% do sexo feminino e 51 (65,4% do masculino. Os resultados apontaram para uma alta correlação entre os testes (r = 0,80, o que permite afirmar que medem o mesmo construto. Apesar disso, a hipótese de que o G36 estaria mais correlacionado com o Fator 2 do R-1 não pôde ser confirmada. Esse dado pode indicar que o G36 não seja unidimensional e se relacione mais com a inteligência cristalizada do que com o fator g.Considering the relevance of obtaining trustful measurement for the evaluation of intelligence, this study aimed at verifying the convergent validity between the R1-Forma B and the G36 tests. 78 students of both gender (34,6% female and 65,4% male, aged from 15 to 64 years old (mean = 36,84; SD = 10,10 and attending to the high school Telecurso were studied. The results indicated a high correlation between tests (r = 0.80, what allowed for the statement that they measure similar constructs. Nevertheless, the hypothesis that G36 would be more correlated with Factor 2 of R-1 could not be confirmed. This information suggests that the G36 is not unidimensional and is more related to crystallized intelligence than the g factor.

  9. Common genetic variations in the CYP2R1 and GC genes are determinants of vitamin D status in Danes

    Nissen, Ioanna

    ), after 6 months intake of vitamin D3-fortified bread and milk (paper II) and in 92 participants in the VitDgen study after artificial UVB irradiation during winter (paper III). Common genetic variations in the CYP2R1 and GC genes were found to be important determinants of vitamin D status in three out...... by genetic variation in vitamin D modulating genes. Twin and family-based studies indicate that genetic variation may have an appreciable influence on vitamin D status. Moreover, several candidate gene studies including two genome-wide association studies (GWAS) have found single nucleotide polymorphisms...... (SNPs) in CYP2R1, CYP24A1, CYP27B1, C10orf88, DHCR7/NADSYN1, GC and VDR genes to be associated with vitamin D status. The main hypothesis of this work was that genetically determined variation in vitamin D metabolism would influence the effect of vitamin D sources (vitamin D...

  10. Comparative molecular field analysis of quinoline derivatives as selective and noncompetitive mGluR1 antagonists.

    Sekhar, Y Nataraja; Nayana, M Ravi Shashi; Ravikumar, Muttineni; Mahmood, S K


    A 3D- QSAR model os Comparative Molecular Field Analysib (CoMFA) of 45 quinoline derivatives as metaborropic glutamate receptor subtype 1 (mGluR1) inhibitors wew investigated. The CoMFA analysis provided a model with q(2) value of 0.827 and r(2) value of 0.990, in which q(2) value of 0.827 and an r(2) value of 0.990, in which the good correlation between the inhibitory activities and the steric and electrostatic molecular field around the analoques was observed. The predictive ability of the models was validated using the set of 12 compounds that were not included in the training set of 33 compounds. These results provided further understanding of the relationship between the structural features of quinolone derivatives and its activities, which should be applicable to design and find new potential mGluR1 inhibitors.

  11. International Conference on Harmonisation; guidance on Q8(R1) Pharmaceutical Development; addition of annex; availability. Notice.


    The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled "Q8(R1) Pharmaceutical Development." The guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The ICH Q8(R1) guidance includes the previously published parent guidance entitled "Q8 Pharmaceutical Development" (Q8 parent guidance) (71 FR 29344; May 22, 2006) and a newly added annex. The annex provides further clarification of key concepts outlined in the Q8 parent guidance and describes the principles of quality by design (QbD). The annex is intended to show how concepts and tools (e.g., design space) outlined in the Q8 parent guidance could be put into practice by the applicant for all dosage forms.

  12. Identificación de Genes R1 y R2 que confieren resistencia a Phytophthora infestans en genotipos colombianos de papa Identification of R1 and R2 Genes conferring resistance to Phytophthora infestans in Colombian potato genotypes

    Núñez Víctor M.


    Full Text Available En Colombia, actualmente existen genotipos de papa con excelente calidad industrial pero muy susceptibles a P. infestans. La mejor manera de combatir este problema es mediante resistencia genética, puesto que la inversión para controlar esta enfermedad por medios químicos es muy costosa, sin olvidar la contaminación ambiental que producen. El objetivo de este trabajo fue la identificación de genes R1 y R2 en los diferenciales de papa respectivos (Solanum tuberosum ssp. tuberosum mediante evaluación de resistencia a P. infestans y la detección molecular por medio de PCR (alelo R1 y AFLP (alelo R2. Para la detección del alelo R1 fueron empleados los primers GP179, GP21, 76-2SF2/76-2SR, SPUD237 y Sol 2749-2770F / Sol 3246-3267R. Los primers GP179, GP21 y SPUD237 fueron inespecíficos para Rl, ya que se generó un producto de amplificación en los diferenciales 1 y 2, así como también en Solanum phureja. Los primers 76-2SF2/76-2SR, y Sol 2749-2770F / Sol 3246-3267R generaron un producto de amplificación en los diferenciales 1 y 2; por el contrario, el fragmento estuvo ausente en el material susceptible. Para la detección del alelo R2, fueron implementados cinco marca­dores AFLP, de los cuales sólo dos fueron reconocidos visualmente en el diferencial 2. Los resultados mostra­ron una evidente correspondencia fenotípica y genotípica con respecto a la presencia de los genes Rl y R2. La identificación molecular de genes de resistencia a P. infestans permitirá desarrollar programas de mejoramiento genético que beneficien directamente los rendimientos de los cultivos de papa, sobre todo los de mayor interés industrial para nuestro país.Excellent industrial quality potato genotypes are currently available in Colombia; however, they are very sus­ceptible to P. infestans. The best way of fighting this problem is by genetic resistance, given that the expense of controlling this disease through chemicals is high, plus the environmental

  13. Galanin Protects from Caspase-8/12-initiated Neuronal Apoptosis in the Ischemic Mouse Brain via GalR1

    Li, Yun; Mei, Zhu; Liu, Shuiqiao; Wang, Tong; Li, Hui; Li, Xiao-Xiao; Han, Song; Yang, Yutao; Li, Junfa; Xu, Zhi-Qing David


    Galanin (GAL) plays key role in many pathophysiological processes, but its role in ischemic stroke remains unclear. Here, the models of 1 h middle cerebral artery occlusion (MCAO)/1-7 d reperfusion (R)-induced ischemic stroke and in vitro cell ischemia of 1 h oxygen-glucose deprivation (OGD)/24 h reoxygenation in primary cultured cortical neurons were used to explore GAL’s effects and its underlying mechanisms. The results showed significant increases of GAL protein levels in the peri-infarct region (P) and infarct core (I) within 48 h R of MCAO mice (p<0.001). The RT-qPCR results also demonstrated significant increases of GAL mRNA during 24-48 h R (p<0.001), and GAL receptors GalR1-2 (but not 3) mRNA levels in the P region at 24 h R of MCAO mice (p<0.001). Furthermore, the significant decrease of infarct volume (p<0.05) and improved neurological outcome (p<0.001-0.05) were observed in MCAO mice following 1 h pre- or 6 h post-treatment of GAL during 1-7 d reperfusion. GalR1 was confirmed as the receptor responsible for GAL-induced neuroprotection by using GalR2/3 agonist AR-M1896 and Lentivirus-based RNAi knockdown of GalR1. GAL treatment inhibited Caspase-3 activation through the upstream initiators Capsases-8/-12 (not Caspase-9) in both P region and OGD-treated cortical neurons. Meanwhile, GAL’s neuroprotective effect was not observed in cortical neurons from conventional protein kinase C (cPKC) γ knockout mice. These results suggested that exogenous GAL protects the brain from ischemic injury by inhibiting Capsase-8/12-initiated apoptosis, possibly mediated by GalR1 via the cPKCγ signaling pathway. PMID:28203483

  14. General expressions for R1ρ relaxation for N-site chemical exchange and the special case of linear chains

    Koss, Hans; Rance, Mark; Palmer, Arthur G.


    Exploration of dynamic processes in proteins and nucleic acids by spin-locking NMR experiments has been facilitated by the development of theoretical expressions for the R1ρ relaxation rate constant covering a variety of kinetic situations. Herein, we present a generalized approximation to the chemical exchange, Rex, component of R1ρ for arbitrary kinetic schemes, assuming the presence of a dominant major site population, derived from the negative reciprocal trace of the inverse Bloch-McConnell evolution matrix. This approximation is equivalent to first-order truncation of the characteristic polynomial derived from the Bloch-McConnell evolution matrix. For three- and four-site chemical exchange, the first-order approximations are sufficient to distinguish different kinetic schemes. We also introduce an approach to calculate R1ρ for linear N-site schemes, using the matrix determinant lemma to reduce the corresponding 3N × 3N Bloch-McConnell evolution matrix to a 3 × 3 matrix. The first- and second order-expansions of the determinant of this 3 × 3 matrix are closely related to previously derived equations for two-site exchange. The second-order approximations for linear N-site schemes can be used to obtain more accurate approximations for non-linear N-site schemes, such as triangular three-site or star four-site topologies. The expressions presented herein provide powerful means for the estimation of Rex contributions for both low (CEST-limit) and high (R1ρ-limit) radiofrequency field strengths, provided that the population of one state is dominant. The general nature of the new expressions allows for consideration of complex kinetic situations in the analysis of NMR spin relaxation data.

  15. Circulating miR-1 as a potential biomarker of doxorubicin-induced cardiotoxicity in breast cancer patients

    Oliveira-Carvalho Vagner, Rigaud; Ferreira, Ludmila R.P; Ayub-Ferreira, Silvia M; Ávila, Mônica S; Brandão, Sara M.G; Cruz, Fátima D; Santos, Marília H.H; Cruz, Cecilia B.B.V; Alves, Marco S.L; Issa, Victor S; Guimarães, Guilherme V; Cunha-Neto, Edécio; Bocchi, Edimar A


    Cardiotoxicity is associated with the chronic use of doxorubicin leading to cardiomyopathy and heart failure. Identification of cardiotoxicity-specific miRNA biomarkers could provide clinicians with a valuable prognostic tool. The aim of the study was to evaluate circulating levels of miRNAs in breast cancer patients receiving doxorubicin treatment and to correlate with cardiac function. This is an ancillary study from “Carvedilol Effect on Chemotherapy-induced Cardiotoxicity” (CECCY trial), which included 56 female patients (49.9±3.3 years of age) from the placebo arm. Enrolled patients were treated with doxorubicin followed by taxanes. cTnI, LVEF, and miRNAs were measured periodically. Circulating levels of miR-1, -133b, -146a, and -423-5p increased during the treatment whereas miR-208a and -208b were undetectable. cTnI increased from 6.6±0.3 to 46.7±5.5 pg/mL (p<0.001), while overall LVEF tended to decrease from 65.3±0.5 to 63.8±0.9 (p=0.053) over 12 months. Ten patients (17.9%) developed cardiotoxicity showing a decrease in LVEF from 67.2±1.0 to 58.8±2.7 (p=0.005). miR-1 was associated with changes in LVEF (r=-0.531, p<0.001). In a ROC curve analysis miR-1 showed an AUC greater than cTnI to discriminate between patients who did and did not develop cardiotoxicity (AUC = 0.851 and 0.544, p= 0.0016). Our data suggest that circulating miR-1 might be a potential new biomarker of doxorubicin-induced cardiotoxicity in breast cancer patients. PMID:28052002

  16. Feasibility studies of producing {sup 99} Mo by capture in the IEA-R1 research reactor

    Concilio, Roberta; Mendonca, Arlindo Gilson; Maiorino, Jose Rubens [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail:;


    Everyday the production of {sup 99} Mo for {sup 99m} Tc generators, becomes more necessary, whose properties are ideal for medical diagnosis. This works presents a description and an analysis of the production of {sup 99} Mo by radioactive capture at {sup 98} Mo using the research reactor IEA-R1 in 5 MW and operating 5 days a week, referring to the use of targets, separation methods, total and specific activity attained and its limitations. (author)

  17. Neutronic parameters characterization of the TRIGA IPR-R1 using scale 6.0 (KENO VI)

    Faria, Victor; Miro, Rafael; Verdu, Gumersindo; Barrachina, Teresa [Institute for Industrial, Radiophysical and Environmental Safety (ISIRYM), Universitat Politecnica de València (Spain); Silva, Clarysson A. Mello da; Pereira, Claubia [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Departamento de Engenharia Nuclear; Dalle, Hugo Moura [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)


    KENO-VI is a Monte Carlo based transport code used to obtain the criticality of a nuclear system. A model built using this code in the SCALE6.0 software system was developed for the characterization of neutronic parameters of the IPR-R1 TRIGA research reactor. A comparison with experimental values and those calculated with a MCNP code model could be then attained with the purpose to validate this methodology. (author)

  18. Can the new RCP R0/R1 classification predict the clinical outcome in ductal adenocarcinoma of the pancreatic head?

    Janot, M S; Kersting, S; Belyaev, O; Matuschek, A; Chromik, A M; Suelberg, D; Uhl, W; Tannapfel, A; Bergmann, U


    According to the International Union Against Cancer (UICC), R1 is defined as the microscopic presence of tumor cells at the surface of the resection margin (RM). In contrast, the Royal College of Pathologists (RCP) suggested to declare R1 already when tumor cells are found within 1 mm of the RM. The aim of this study was to determine the significance of the RM concerning the prognosis of pancreatic ductal adenocarcinoma (PDAC). From 2007 to 2009, 62 patients underwent a curative operation for PDAC of the pancreatic head. The relevance of R status on cumulative overall survival (OS) was assessed on univariate and multivariate analysis for both the classic R classification (UICC) and the suggestion of the RCP. Following the UICC criteria, a positive RM was detected in 8 %. Along with grading and lymph node ratio, R status revealed a significant impact on OS on univariate and multivariate analysis. Applying the suggestion of the RCP, R1 rate rose to 26 % resulting in no significant impact on OS in univariate analysis. Our study has shown that the RCP suggestion for R status has no impact on the prognosis of PDAC. In contrast, our data confirmed the UICC R classification of RM as well as N category, grading, and lymph node ratio as significant prognostic factors.

  19. Synapse-specific mGluR1-dependent long-term potentiation in interneurones regulates mouse hippocampal inhibition

    Lapointe, Valérie; Morin, France; Ratté, Stéphanie; Croce, Ariane; Conquet, François; Lacaille, Jean-Claude


    Hippocampal CA1 inhibitory interneurones control the excitability and synchronization of pyramidal cells, and participate in hippocampal synaptic plasticity. Pairing theta-burst stimulation (TBS) with postsynaptic depolarization, we induced long-term potentiation (LTP) of putative single-fibre excitatory postsynaptic currents (EPSCs) in stratum oriens/alveus (O/A) interneurones of mouse hippocampal slices. LTP induction was absent in metabotropic glutamate receptor 1 (mGluR1) knockout mice, was correlated with the postsynaptic presence of mGluR1a, and required a postsynaptic Ca2+ rise. Changes in paired-pulse facilitation and coefficient of variation indicated that LTP expression involved presynaptic mechanisms. LTP was synapse specific, occurring selectively at synapses modulated by presynaptic group II, but not group III, mGluRs. Furthermore, the TBS protocol applied in O/A induced a long-term increase of polysynaptic inhibitory responses in CA1 pyramidal cells, that was absent in mGluR1 knockout mice. These results uncover the mechanisms of a novel form of interneurone synaptic plasticity that can adaptively regulate inhibition of hippocampal pyramidal cells. PMID:14673190

  20. Bacterial mitosis: ParM of plasmid R1 moves plasmid DNA by an actin-like insertional polymerization mechanism.

    Møller-Jensen, Jakob; Borch, Jonas; Dam, Mette; Jensen, Rasmus B; Roepstorff, Peter; Gerdes, Kenn


    Bacterial DNA segregation takes place in an active and ordered fashion. In the case of Escherichia coli plasmid R1, the partitioning system (par) separates paired plasmid copies and moves them to opposite cell poles. Here we address the mechanism by which the three components of the R1 par system act together to generate the force required for plasmid movement during segregation. ParR protein binds cooperatively to the centromeric parC DNA region, thereby forming a complex that interacts with the filament-forming actin-like ParM protein in an ATP-dependent manner, suggesting that plasmid movement is powered by insertional polymerization of ParM. Consistently, we find that segregating plasmids are positioned at the ends of extending ParM filaments. Thus, the process of R1 plasmid segregation in E. coli appears to be mechanistically analogous to the actin-based motility operating in eukaryotic cells. In addition, we find evidence suggesting that plasmid pairing is required for ParM polymerization.

  1. Inhibition of mGluR1 and IP3Rs impairs long-term memory formation in young chicks.

    Baker, K D; Edwards, T M; Rickard, N S


    Calcium (Ca(2+)) is involved in a myriad of cellular functions in the brain including synaptic plasticity. However, the role of intracellular Ca(2+) stores in memory processing remains poorly defined. The current study explored a role for glutamate-dependent intracellular Ca(2+) release in memory processing via blockade of metabotropic glutamate receptor subtype 1 (mGluR1) and inositol (1,4,5)-trisphosphate receptors (IP(3)Rs). Using a single-trial discrimination avoidance task developed for the young chick, administration of the specific and potent mGluR1 antagonist JNJ16259685 (500nM, immediately post-training, ic), or the IP(3)R antagonist Xestospongin C (5microM, immediately post-training, ic), impaired retention from 90min post-training. These findings are consistent with mGluR1 activating IP(3)Rs to release intracellular Ca(2+) required for long-term memory formation and have been interpreted within an LTP2 model. The consequences of different patterns of retention loss following ryanodine receptor (RyR) and IP(3)R inhibition are discussed.

  2. Vitamin D Insufficiency in Arabs and South Asians Positively Associates with Polymorphisms in GC and CYP2R1 Genes

    Elkum, Naser; Alkayal, Fadi; Noronha, Fiona; Ali, Maisa M.; Melhem, Motasem; Al-Arouj, Monira; Bennakhi, Abdullah; Behbehani, Kazem; Alsmadi, Osama; Abubaker, Jehad


    Background A number of genetic studies have reported an association between vitamin D related genes such as group-specific component gene (GC), Cytochrome P450, family 2, subfamily R, polypeptide 1 (CYP2R1) and 7-dehydrocholesterol reductase/nicotinamide-adenine dinucleotide synthetase 1 (DHCR7/NADSYN1) and serum levels of the active form of Vitamin D, 25 (OH) D among African Americans, Caucasians, and Chinese. Little is known about how genetic variations associate with, or contribute to, 25(OH)D levels in Arabs populations. Methods Allele frequencies of 18 SNPs derived from CYP2R1, GC, and DHCR7/NADSYN1 genes in 1549 individuals (Arabs, South Asians, and Southeast Asians living in Kuwait) were determined using real time genotyping assays. Serum levels of 25(OH)D were measured using chemiluminescence immunoassay. Results GC gene polymorphisms (rs17467825, rs3755967, rs2282679, rs7041 and rs2298850) were found to be associated with 25(OH)D serum levels in Arabs and South Asians. Two of the CYP2R1 SNPs (rs10500804 and rs12794714) and one of GC SNPs (rs1155563) were found to be significantly associated with 25(OH)D serum levels only in people of Arab origin. Across all three ethnicities none of the SNPs of DHCR7/NADSYN1 were associated with serum 25(OH)D levels and none of the 18 SNPs were significantly associated with serum 25(OH)D levels in people from South East Asia. Conclusion Our data show for the first time significant association between the GC (rs2282679 and rs7041), CYP2R1 (rs10741657) SNPs and 25(OH)D levels. This supports their roles in vitamin D Insufficiency in Arab and South Asian populations respectively. Interestingly, two of the CYP2R1 SNPs (rs10500804 and rs12794714) and one GC SNP (rs1155563) were found to correlate with vitamin D in Arab population exclusively signifying their importance in this population. PMID:25405862

  3. Characterization of heterosubunit complexes formed by the R1 and R2 subunits of herpes simplex virus 1 and equine herpes virus 4 ribonucleotide reductase.

    Sun, Y; Conner, J


    We report on the separate PCR cloning and subsequent expression and purification of the large (R1) and small (R2) subunits from equine herpes virus type 4 (EHV-4) ribonucleotide reductase. The EHV-4 R1 and R2 subunits reconstituted an active enzyme and their abilities to complement the R1 and R2 subunits from the closely related herpes simplex virus 1 (HSV-1) ribonucleotide reductase, with the use of subunit interaction and enzyme activity assays, were analysed. Both EHV-4 R1/HSV-1 R2 and HSV-1 R1/EHV-4 R2 were able to assemble heterosubunit complexes but, surprisingly, neither of these complexes was fully active in enzyme activity assays; the EHV-4 R1/HSV-1 R2 and HSV-1 R1/EHV-4 R2 enzymes had 50% and 5% of their respective wild-type activities. Site-directed mutagenesis was used to alter two non-conserved residues located within the highly conserved and functionally important C-termini of the EHV-4 and HSV-1 R1 proteins. Mutation of Pro-737 to Lys and Lys-1084 to Pro in EHV-4 and HSV-1 R1 respectively had no effects on subunit assembly. Mutation of Pro-737 to Lys in EHV-4 R1 decreased enzyme activity by 50%; replacement of Lys-1084 by Pro in HSV-1 R1 had no effect on enzyme activity. Both alterations failed to restore full enzyme activities to the heterosubunit enzymes. Therefore probably neither of these amino acids has a direct role in catalysis. However, mutation of the highly conserved Tyr-1111 to Phe in HSV-1 R1 inactivated enzyme activity without affecting subunit interaction.

  4. BubR1 Acts as a Promoter in Cellular Motility of Human Oral Squamous Cancer Cells through Regulating MMP-2 and MMP-9

    Chou-Kit Chou


    Full Text Available BubR1 is a critical component of spindle assembly checkpoint, ensuring proper chromatin segregation during mitosis. Recent studies showed that BubR1 was overexpressed in many cancer cells, including oral squamous cell carcinomas (OSCC. However, the effect of BubR1 on metastasis of OSCC remains unclear. This study aimed to unravel the role of BubR1 in the progression of OSCC and confirm the expression of BubR1 in a panel of malignant OSCC cell lines with different invasive abilities. The results of quantitative real-time PCR showed that the mRNA level of BubR1 was markedly increased in four OSCC cell lines, Ca9-22, HSC3, SCC9 and Cal-27 cells, compared to two normal cells, normal human oral keratinocytes (HOK and human gingival fibroblasts (HGF. Moreover, the expression of BubR1 in these four OSCC cell lines was positively correlated with their motility. Immunofluorescence revealed that BubR1 was mostly localized in the cytosol of human gingival carcinoma Ca9-22 cells. BubR1 knockdown significantly decreased cellular invasion but slightly affect cellular proliferation on both Ca9-22 and Cal-27 cells. Consistently, the activities of metastasis-associated metalloproteinases MMP-2 and MMP-9 were attenuated in BubR1 knockdown Ca9-22 cells, suggesting the role of BubR1 in promotion of OSCC migration. Our present study defines an alternative pathway in promoting metastasis of OSCC cells, and the expression of BubR1 could be a prognostic index in OSCC patients.

  5. The C5a/C5aR1 axis controls the development of experimental allergic asthma independent of LysM-expressing pulmonary immune cells.

    Wiese, Anna V; Ender, Fanny; Quell, Katharina M; Antoniou, Konstantina; Vollbrandt, Tillman; König, Peter; Köhl, Jörg; Laumonnier, Yves


    C5a regulates the development of maladaptive immune responses in allergic asthma mainly through the activation of C5a receptor 1 (C5aR1). Yet, the cell types and the mechanisms underlying this regulation are ill-defined. Recently, we described increased C5aR1 expression in lung tissue eosinophils but decreased expression in airway and pulmonary macrophages as well as in pulmonary CD11b+ conventional dendritic cells (cDCs) and monocyte-derived DCs (moDCs) during the allergic effector phase using a floxed green fluorescent protein (GFP)-C5aR1 knock-in mouse. Here, we determined the role of C5aR1 signaling in neutrophils, moDCs and macrophages for the pulmonary recruitment of such cells and the importance of C5aR1-mediated activation of LysM-expressing cells for the development of allergic asthma. We used LysM-C5aR1 KO mice with a specific deletion of C5aR1 in LysMCre-expressing cells and confirmed the specific deletion of C5aR1 in neutrophils, macrophages and moDCs in the airways and/or the lung tissue. We found that alveolar macrophage numbers were significantly increased in LysM-C5aR1 KO mice. Induction of ovalbumin (OVA)-driven experimental allergic asthma in GFP-C5aR1fl/fl and LysM-C5aR1 KO mice resulted in strong but similar airway resistance, mucus production and Th2/Th17 cytokine production. In contrast, the number of airway but not of pulmonary neutrophils was lower in LysM-C5aR1 KO as compared with GFP-C5aR1fl/fl mice. The recruitment of macrophages, cDCs, moDCs, T cells and type 2 innate lymphoid cells was not altered in LysM-C5aR1 KO mice. Our findings demonstrate that C5aR1 is critical for steady state control of alveolar macrophage numbers and the transition of neutrophils from the lung into the airways in OVA-driven allergic asthma. However, C5aR1 activation of LysM-expressing cells plays a surprisingly minor role in the recruitment and activation of such cells and the development of the allergic phenotype in OVA-driven experimental allergic asthma.

  6. Muscle-specific MicroRNA1 (miR1) Targets Heat Shock Protein 70 (HSP70) during Dexamethasone-mediated Atrophy*

    Kukreti, Himani; Amuthavalli, Kottaiswamy; Harikumar, Arigela; Sathiyamoorthy, Sushmitha; Feng, Peng Zhao; Anantharaj, Rengaraj; Tan, Suan Liang Kelvin; Lokireddy, Sudarsanareddy; Bonala, Sabeera; Sriram, Sandhya; McFarlane, Craig; Kambadur, Ravi; Sharma, Mridula


    High doses of dexamethasone (Dex) or myostatin (Mstn) induce severe atrophy of skeletal muscle. Here we show a novel microRNA1 (miR1)-mediated mechanism through which Dex promotes skeletal muscle atrophy. Using both C2C12 myotubes and mouse models of Dex-induced atrophy we show that Dex induces miR1 expression through glucocorticoid receptor (GR). We further show that Mstn treatment facilitates GR nuclear translocation and thereby induces miR1 expression. Inhibition of miR1 in C2C12 myotubes attenuated the Dex-induced increase in atrophy-related proteins confirming a role for miR1 in atrophy. Analysis of miR1 targets revealed that HSP70 is regulated by miR1 during atrophy. Our results demonstrate that increased miR1 during atrophy reduced HSP70 levels, which resulted in decreased phosphorylation of AKT, as HSP70 binds to and protects phosphorylation of AKT. We further show that loss of pAKT leads to decreased phosphorylation, and thus, enhanced activation of FOXO3, up-regulation of MuRF1 and Atrogin-1, and progression of skeletal muscle atrophy. Based on these results, we propose a model whereby Dex- and Mstn-mediated atrophic signals are integrated through miR1, which then either directly or indirectly, inhibits the proteins involved in providing protection against atrophy. PMID:23297411

  7. The architecture of the BubR1 tetratricopeptide tandem repeat defines a protein motif underlying mitotic checkpoint-kinetochore communication

    Bolanos-Garcia, Victor M; Nilsson, Jakob; Blundell, Tom L


    advance to anaphase before every chromosome is properly attached to microtubules of the mitotic spindle. The architecture of the KNL1-BubR1 complex reveals important features of the molecular recognition between SAC components and the kinetochore. The interaction is important for a functional SAC...... as substitution of BubR1 residues engaged in KNL1 binding impaired the SAC and BubR1 recruitment into checkpoint complexes in stable cell lines. Here we discuss the implications of the disorder-to-order transition of KNL1 upon BubR1 binding for SAC signaling and propose a mechanistic model of how BUBs binding may...

  8. Identification and characterization of EctR1, a new transcriptional regulator of the ectoine biosynthesis genes in the halotolerant methanotroph Methylomicrobium alcaliphilum 20Z.

    Mustakhimov, Ildar I; Reshetnikov, Alexander S; Glukhov, Anatoly S; Khmelenina, Valentina N; Kalyuzhnaya, Marina G; Trotsenko, Yuri A


    Genes encoding key enzymes of the ectoine biosynthesis pathway in the halotolerant obligate methanotroph Methylomicrobium alcaliphilum 20Z have been shown to be organized into an ectABC-ask operon. Transcription of the ect operon is initiated from two promoters, ectAp(1) and ectAp(2) (ectAp(1)p(2)), similar to the sigma(70)-dependent promoters of Escherichia coli. Upstream of the gene cluster, an open reading frame (ectR1) encoding a MarR-like transcriptional regulator was identified. Investigation of the influence of EctR1 on the activity of the ectAp(1)p(2) promoters in wild-type M. alcaliphilum 20Z and ectR1 mutant strains suggested that EctR1 is a negative regulator of the ectABC-ask operon. Purified recombinant EctR1-His(6) specifically binds as a homodimer to the putative -10 motif of the ectAp(1) promoter. The EctR1 binding site contains a pseudopalindromic sequence (TATTTAGT-GT-ACTATATA) composed of 8-bp half-sites separated by 2 bp. Transcription of the ectR1 gene is initiated from a single sigma(70)-like promoter. The location of the EctR1 binding site between the transcriptional and translational start sites of the ectR1 gene suggests that EctR1 may regulate its own expression. The data presented suggest that in Methylomicrobium alcaliphilum 20Z, EctR1-mediated control of the transcription of the ect genes is not the single mechanism for the regulation of ectoine biosynthesis.

  9. Ageing implementation and refurbishment development at the IEA-R1 nuclear research reactor: a 15 years experience

    Cardenas, Jose Patricio N.; Ricci Filho, Walter; Carvalho, Marcos R. de; Berretta, Jose Roberto; Marra Neto, Adolfo, E-mail: ahiru@ipen.b, E-mail: wricci@ipen.b, E-mail: carvalho@ipen.b, E-mail: jrretta@ipen.b, E-mail: amneto@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    IPEN (Instituto de Pesquisas Energeticas e Nucleares) is a nuclear research center established into the Secretary of Science and Technology from the government of the state of Sao Paulo, and administered both technically and financially by Comissao Nacional de Energia Nuclear (CNEN), a federal government organization under the Ministry of Science and Technology. The institute is located inside the campus of the University of Sao Paulo, Sao Paulo city, Brazil. One of major nuclear facilities at IPEN is the IEA-R1 nuclear research reactor. It is the unique Brazilian research reactor with substantial power level suitable for application with research in physics, chemistry, biology and engineering, as well as radioisotope production for medical and other applications. Designed and built by Babcok-Wilcox, in accordance with technical specifications established by the Brazilian Nuclear Energy Commission, and financed by the US Atoms for Peace Program, it is a swimming pool type reactor, moderated and cooled by light water and uses graphite and beryllium as reflector elements. The first criticality was achieved on September 16, 1957 and the reactor is currently operating at 4.0 MW on a 64h per week cycle. Since 1996, an IEA-R1 reactor ageing study was established at the Research Reactor Center (CRPq) related with general deterioration of components belonging to some operational systems, as cooling towers from secondary cooling system, piping and pumps, sample irradiation devices, radiation monitoring system, fuel elements, rod drive mechanisms, nuclear and process instrumentation and safety operational system. Although basic structures are almost the same as the original design, several improvements and modifications in components, systems and structures had been made along reactor life. This work aims to show the development of the ageing program in the IEA-R1 reactor and the upgrading (modernization) that was carried out, concerning several equipment and system in the

  10. Near-infrared Spectroscopic Observations of Comet C/2013 R1 (Lovejoy) by WINERED: CN Red-system Band Emission

    Shinnaka, Yoshiharu; Kawakita, Hideyo; Kondo, Sohei; Ikeda, Yuji; Kobayashi, Naoto; Hamano, Satoshi; Sameshima, Hiroaki; Fukue, Kei; Matsunaga, Noriyuki; Yasui, Chikako; Izumi, Natsuko; Mizumoto, Misaki; Otsubo, Shogo; Takenaka, Keiichi; Watase, Ayaka; Kawanishi, Takafumi; Nakanishi, Kenshi; Nakaoka, Tetsuya


    Although high-resolution spectra of the CN red-system band are considered useful in cometary sciences, e.g., in the study of isotopic ratios of carbon and nitrogen in cometary volatiles, there have been few reports to date due to the lack of high-resolution (R ≡ λ/Δλ > 20,000) spectrographs in the near-infrared region around ˜1 μm. Here, we present the high-resolution emission spectrum of the CN red-system band in comet C/2013 R1 (Lovejoy), acquired by the near-infrared high-resolution spectrograph WINERED mounted on the 1.3 m Araki telescope at the Koyama Astronomical Observatory, Kyoto, Japan. We applied our fluorescence excitation models for CN, based on modern spectroscopic studies, to the observed spectrum of comet C/2013 R1 (Lovejoy) to search for CN isotopologues (13C14N and 12C15N). We used a CN fluorescence excitation model involving both a “pure” fluorescence excitation model for the outer coma and a “fully collisional” fluorescence excitation model for the inner coma region. Our emission model could reproduce the observed 12C14N red-system band of comet C/2013 R1 (Lovejoy). The derived mixing ratio between the two excitation models was 0.94(+0.02/-0.03):0.06(+0.03/-0.02), corresponding to the radius of the collision-dominant region of ˜800-1600 km from the nucleus. No isotopologues were detected. The observed spectrum is consistent, within error, with previous estimates in comets of 12C/13C (˜90) and 14N/15N (˜150).

  11. High-dispersion spectroscopic observations of comet C/2013 R1 (Lovejoy) with the Subaru Telescope on 2013 November 15

    Shinnaka, Y.; Kawakita, H.; Nagashima, M.; Kobayashi, H.; Decock, A.; Jehin, E.


    Comet C/2013 R1 (Lovejoy) probably originates from the Oort Cloud. Comet Lovejoy provided us with great opportunities to investigate the chemical composition of the comet thanks to its brightness and elongation angle from October 2013 to March 2014. We observed comet C/2013 R1 (Lovejoy) on 2013 November 15 UT using the High Dispersion Spectrograph (HDS) mounted on the Subaru Telescope on Mauna Kea. Its heliocentric and geocentric distances were 1.066 and 0.412 au, respectively. The obtained spectra cover the wavelength region from 360 to 830 nm with the resolving power of R = λ/Δλ = 72,000 for the slit size of 0''.5 × 9''.0 (360-520 nm) and 0''.5 × 9''.0 (550-830 nm) on the sky. Exposure time was 130 minutes in total (100 minutes for shorter and 30 minutes for longer wavelength regions). We could identify many species such as the radicals (CN, CH, C_{3}, C_{2}, NH_{2}, etc.), ions (CH^{+}, H_{2}O^{+}), and atoms ([OI] and NaI) in the spectra and many unidentified lines were also detected. Here we discuss the chemical reaction in cometary coma and the origin of icy materials of comet C/2013 R1 (Lovejoy), based on the high-dispersion spectra in the optical wavelength regions. We present (1) the ortho-to-para abundance ratios (OPRs) of water and ammonia inferred from the high-dispersion spectra of H_{2}O^{+} and NH_{2}, (2) the green-to-red line ratio of forbidden oxygen emissions, (3) the nitrogen and the carbon isotopic ratios of CN, and (4) spatial distributions of radicals, atoms, and dust continuum in the inner coma.

  12. Differential expression of ghrelin and its receptor (GHS-R1a) in various adrenal tumors and normal adrenal gland.

    Ueberberg, B; Unger, N; Sheu, S Y; Walz, M K; Schmid, K W; Saeger, W; Mann, K; Petersenn, S


    Ghrelin is a newly characterized, widely distributed peptide thought to be involved in the regulation of appetite. Significant effects on the release of growth hormone (GH) and ACTH have been demonstrated. This study compares the expression of ghrelin and its receptor (GHS-R) in various adrenal tumors and normal adrenal gland. Normal adrenal tissue was obtained after autopsy. Tissue was obtained from 13 pheochromocytomas (PHEOs), 15 cortisol-secreting adenomas (CPAs), 12 aldosterone-secreting adenomas (APAs), and 16 nonfunctional adenomas (NFAs) following laparoscopic surgery. Expression of ghrelin and GHS-R1a was investigated on RNA levels by using real-time reverse transcription polymerase chain reaction (RT-PCR) and on protein levels by using immunohistochemistry. In the seven normal adrenal glands analyzed, ghrelin mRNA levels were 12-fold lower than in stomach. Ghrelin protein expression was confirmed by immunohistochemistry. In all adrenal tumors, relevant levels of ghrelin mRNA were observed, with significantly lower expression in PHEOs and APAs than in normal adrenal gland. Ghrelin protein was detected in 0% of PHEOs, 55% of APAs, 87% of CPAs, and 54% of NFAs. GHS-R1a mRNA expression was detectable in normal adrenal gland, but the receptor protein was absent. In adrenal tumors, detectable levels of receptor mRNA were found in 38% of PHEOs, 13% of CPAs, and 25% of NFAs. GHS-R1a protein was absent in the majority of adrenal tumors. Expression of ghrelin in normal adrenal gland and adrenal tumors may indicate some unknown physiological function. The pathophysiological relevance of ghrelin expression in adrenal tumors remains to be investigated.

  13. Application of nondestructive methods for qualification of high density fuels in the IEA-R1 reactor

    Silva, Jose E.R.; Silva, Antonio T.; Domingos, Douglas B.; Terremoto, Luis A.A., E-mail: jersilva@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    IPEN/CNEN-SP manufactures fuels to be used in its research reactor - the IEA-R1. To qualify those fuels, it is necessary to check if they have a good performance under irradiation. As Brazil still does not have nuclear research reactors with high neutron fluxes, or suitable hot cells for carrying out post-irradiation examination of nuclear fuels, IPEN/CNEN-SP has conducted a fuel qualification program based on the use of uranium compounds (U{sub 3}O{sub 8} and U{sub 3}Si{sub 2} dispersed in Al matrix) internationally tested and qualified to be used in research reactors, and has attained experience in the technological development stages for the manufacturing of fuel plates, irradiation and non-destructive post-irradiation testing. Fuel elements containing low volume fractions of fuel in the dispersion were manufactured and irradiated successfully directly in the core of the IEA-R1. However, there are plans at IPEN/CNEN-SP to increase the uranium density of the fuels. Ten fuel miniplates (five containing U{sub 3}O{sub 8}-Al and five containing U{sub 3}Si{sub 2}-Al), with densities of 3.2 gU/cm{sup 3} and 4.8 gU/cm{sup 3} respectively, are being irradiated inside an irradiation device placed in a peripheral position of the IEA-R1 core. Non-destructive methods will be used to evaluate irradiation performance of the fuel miniplates after successive cycles of irradiation, by means: monitoring the reactor parameters during operation; periodic underwater visual inspection of fuel miniplates, eventual sipping test for fuel miniplates suspected of leakage and underwater measuring of the miniplate thickness for assessment of the fuel miniplate swelling. (author)

  14. A systematic study of gene mutations in urothelial carcinoma; inactivating mutations in TSC2 and PIK3R1.

    Gottfrid Sjödahl

    Full Text Available BACKGROUND: Urothelial carcinoma (UC is characterized by frequent gene mutations of which activating mutations in FGFR3 are the most frequent. Several downstream targets of FGFR3 are also mutated in UC, e.g., PIK3CA, AKT1, and RAS. Most mutation studies of UCs have been focused on single or a few genes at the time or been performed on small sample series. This has limited the possibility to investigate co-occurrence of mutations. METHODOLOGY/PRINCIPAL FINDINGS: We performed mutation analyses of 16 genes, FGFR3, PIK3CA, PIK3R1 PTEN, AKT1, KRAS, HRAS, NRAS, BRAF, ARAF, RAF1, TSC1, TSC2, APC, CTNNB1, and TP53, in 145 cases of UC. We show that FGFR3 and PIK3CA mutations are positively associated. In addition, we identified PIK3R1 as a target for mutations. We demonstrate a negative association at borderline significance between FGFR3 and RAS mutations, and show that these mutations are not strictly mutually exclusive. We show that mutations in BRAF, ARAF, RAF1 rarely occurs in UC. Our data emphasize the possible importance of APC signaling as 6% of the investigated tumors either showed inactivating APC or activating CTNNB1 mutations. TSC1, as well as TSC2, that constitute the mTOR regulatory tuberous sclerosis complex were found to be mutated at a combined frequency of 15%. CONCLUSIONS/SIGNIFICANCE: Our data demonstrate a significant association between FGFR3 and PIK3CA mutations in UC. Moreover, the identification of mutations in PIK3R1 further emphasizes the importance of the PI3-kinase pathway in UC. The presence of TSC2 mutations, in addition to TSC1 mutations, underlines the involvement of mTOR signaling in UC.

  15. Partial recessive IFN-γR1 deficiency: genetic, immunological and clinical features of 14 patients from 11 kindreds

    Sologuren, Ithaisa; Boisson-Dupuis, Stéphanie; Pestano, Jose; Vincent, Quentin Benoit; Fernández-Pérez, Leandro; Chapgier, Ariane; Cárdenes, María; Feinberg, Jacqueline; García-Laorden, M. Isabel; Picard, Capucine; Santiago, Esther; Kong, Xiaofei; Jannière, Lucile; Colino, Elena; Herrera-Ramos, Estefanía; Francés, Adela; Navarrete, Carmen; Blanche, Stéphane; Faria, Emilia; Remiszewski, Paweł; Cordeiro, Ana; Freeman, Alexandra; Holland, Steven; Abarca, Katia; Valerón-Lemaur, Mónica; Gonçalo-Marques, José; Silveira, Luisa; García-Castellano, José Manuel; Caminero, José; Pérez-Arellano, José Luis; Bustamante, Jacinta; Abel, Laurent; Casanova, Jean-Laurent; Rodríguez-Gallego, Carlos


    We report a series of 14 patients from 11 kindreds with recessive partial (RP)-interferon (IFN)-γR1 deficiency. The I87T mutation was found in nine homozygous patients from Chile, Portugal and Poland, and the V63G mutation was found in five homozygous patients from the Canary Islands. Founder effects accounted for the recurrence of both mutations. The most recent common ancestors of the patients with the I87T and V63G mutations probably lived 1600 (875–2950) and 500 (200–1275) years ago, respectively. The two alleles confer phenotypes that are similar but differ in terms of IFN-γR1 levels and residual response to IFN-γ. The patients suffered from bacillus Calmette-Guérin-osis (n= 6), environmental mycobacteriosis (n= 6) or tuberculosis (n= 1). One patient did not suffer from mycobacterial infections but had disseminated salmonellosis, which was also present in two other patients. Age at onset of the first environmental mycobacterial disease differed widely between patients, with a mean value of 11.25 ± 9.13 years. Thirteen patients survived until the age of 14.82 ± 11.2 years, and one patient died at the age of 7 years, 9 days after the diagnosis of long-term Mycobacterium avium infection and the initiation of antimycobacterial treatment. Up to 10 patients are currently free of infection with no prophylaxis. The clinical heterogeneity of the 14 patients was not clearly related to either IFNGR1 genotype or the resulting cellular phenotype. RP-IFN-γR1 deficiency is, thus, more common than initially thought and should be considered in both children and adults with mild or severe mycobacterial diseases. PMID:21266457

  16. Circuits design of action logics of the protection system of nuclear reactor IAN-R1 of Colombia; Diseno de los circuitos de la logica de actuacion del sistema de proteccion del reactor nuclear IAN-R1 de Colombia

    Gonzalez M, J. L.; Rivero G, T.; Sainz M, E., E-mail: [ININ, Carretera Mexico-Toluca s/n, 52750 Ocoyoacac, Estado de Mexico (Mexico)


    Due to the obsolescence of the instrumentation and control system of the nuclear research reactor IAN-R1, the Institute of Geology and Mining of Colombia, IngeoMinas, launched an international convoking for renewal it which was won by the Instituto Nacional de Investigaciones Nucleares (ININ). Within systems to design, the reactor protection system is described as important for safety, because this carried out, among others two primary functions: 1) ensuring the reactor shutdown safely, and 2) controlling the interlocks to protect against operational errors if defined conditions have not been met. To fulfill these functions, the various subsystems related to the safety report the state in which they are using binary signals and are connected to the inputs of two redundant logic wiring circuits called action logics (Al) that are part of the reactor protection system. These Al also serve as logical interface to indicate at all times the status of subsystems, both the operator and other systems. In the event that any of the subsystems indicates a state of insecurity in the reactor, the Al generate signals off (or scram) of the reactor, maintaining the interlock until the operator sends a reset signal. In this paper the design, implementation, verification and testing of circuits that make up the Al 1 and 2 of IAN-R1 reactor is described, considering the fulfillment of the requirements that the different international standards imposed on this type of design. (Author)

  17. Study of molybdenum-99 production in IEA-R1m reactor using a beryllium irradiator; Estudos sobre a producao de molibdenio no reator IEA-R1m utilizando um irradiador de berilio

    Ricci Filho, Walter [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil); Moreira, Joao M.L. [Centro Tecnologico da Marinha (CTMSP), Sao Paulo, SP (Brazil)


    The IEA-R1 reactor has undergone a modernization to increase its operating power to 5 MW, in order to allow a more efficient production of the {sup 99} Mo radioisotope. An irradiation element made of Be was acquired for the reactor and studies were initiated for determining its performance when compared to other irradiators available in the reactor, the water and graphite types. The results obtained showed some advantages of the Be irradiation element for producing {sup 99} Mo: the epithermal neutron flux in the irradiation element is approximately 15% greater than in the water and in the graphite irradiation elements; the negative reactivity introduced in the reactor by the Be irradiation element is substantially smaller than the those introduced by the other elements: - 1636 pcm for the Be irradiator, -2568 for the graphite irradiator and -2977 pcm for the water irradiator. It is concluded that the production of the {sup 99M}o radioisotope with the Be irradiation element can be increased by 15% in the IEA-R1m reactor. It also requires less fuel for the reactor operation due to the smaller negative reactivity introduced in the reactor core. (author) 4 refs., 9 figs., 1 tab.

  18. 三七皂苷 R1壳聚糖纳米粒的制备及其工艺优化%Preparation and process optimization of notoginsenoside R1 chitosan nanoparticles

    虞璐; 魏宇峰


    目的:制备三七皂苷 R1壳聚糖纳米粒,为该药物的临床应用提供新的给药方式。方法制备三七皂苷 R1壳聚糖纳米粒,采用高效液相色谱法(HPLC)检测纳米粒中三七皂苷 R1的含量,采用正交试验优化纳米粒的制备工艺,并验证优化的纳米粒的制备工艺。结果HPLC 标准曲线方程为 A =911.49C-1803.4(r =0.9999),线性范围为25~900μg/mL。日内精密度分别为1.520%、0.884%、0.969%(n =6);日间精密度分别为1.591%、1.447%、1.269%(n =6)。低、中、高浓度的回收率分别为(98.11±1.16)%、(101.27±0.59)%、(100.97±0.82)%。正交试验的4个因素:壳聚糖浓度、药物与载体的质量比、温度、转速,每个因素分别选择3个水平。选择平均粒径、包封率、载药量为控制指标,使用综合加权评分法确定试验结果,按照 L9(34)正交表设计正交试验。优化的工艺为:壳聚糖浓度2.0%,药物与载体的重量比例20%,温度35℃,转速600 r/min。按照优化的工艺制备纳米粒,平均粒径为(123.40±7.68)nm,包封率为(58.41±1.59)%,载药量为(10.46±0.53)%。结论优化的三七皂苷 R1壳聚糖纳米粒的制备工艺简单易操作,包封率和载药量较高,作为一种新剂型有良好的临床应用前景。%Objective To prepare optimization of notoginsenoside R1 chitosan nanoparticles,to provide a theoretical basis for clinical application of the drug.Methods Notoginsenoside R1 chitosan nanoparticles were prepared,HPLC method was used to detect the content of notoginsenoside R1 chitosan nanoparticles,preparation technology of nanoparticles were optimized by orthogonal experiment,and the optimized preparation technology of nanoparticles was verified.Results HPLC standard curve equation was A =911.49C -1803.4(r =0.999 9),linear range was from 25 to 900 g

  19. Reactivity-worth estimates of the OSMOSE samples in the MINERVE reactor R1-UO2 configuration.

    Klann, R. T.; Perret, G.; Nuclear Engineering Division


    An initial series of calculations of the reactivity-worth of the OSMOSE samples in the MINERVE reactor with the R1-UO2 core configuration were completed. The reactor model was generated using the REBUS code developed at Argonne National Laboratory. The calculations are based on the specifications for fabrication, so they are considered preliminary until sampling and analysis have been completed on the fabricated samples. The estimates indicate a range of reactivity effect from -22 pcm to +25 pcm compared to the natural U sample.

  20. Attenuated Pik3r1 Expression Prevents Insulin Resistance and Adipose Tissue Macrophage Accumulation in Diet-Induced Obese Mice

    McCurdy, Carrie E.; Schenk, Simon; Holliday, Michael J.; Philp, Andrew; Houck, Julie A.; Patsouris, David; MacLean, Paul S.; Majka, Susan M.; Klemm, Dwight J.; Friedman, Jacob E. (Jed)


    Obese white adipose tissue (AT) is characterized by large-scale infiltration of proinflammatory macrophages, in parallel with systemic insulin resistance; however, the cellular stimulus that initiates this signaling cascade and chemokine release is still unknown. The objective of this study was to determine the role of the phosphoinositide 3-kinase (PI3K) regulatory subunits on AT macrophage (ATM) infiltration in obesity. Here, we find that the Pik3r1 regulatory subunits (i.e., p85α/p55α/p50α...

  1. Distribution of mGluR1alpha and SMI 311 immunoreactive Lugaro cells in the kitten cerebellum.

    Víg, Julianna; Takács, József; Vastagh, Csaba; Baldauf, Zsolt; Veisenberger, Eleonóra; Hámori, József


    The Lugaro cell is a feedback interneuron of the cerebellar cortex, recognizable by its characteristic morphology. Postnatal neuronal migration to the cortex has been described for several cerebellar interneurons. Since in our previous studies we observed Lugaro-like cells (LCs) in the white matter (WM) and internal granular layer (IGL) of the cerebellum of young cats, we assumed that a proportion of these cells migrate also postnatally to their destination. In the present study using and immunostaining for the metabotropic glutamate receptor mGluR1alpha and neurofilament protein SMI 311 the number and spatial distribution of LCs at different postnatal days were investigated. We found that the number and distribution of both mGluR1a-immunoreactive (ir) and of SMI 311-ir LCs changed with age in the developing cerebellar cortex of kittens: developing LCs express mGluR1alpha already in the newborn, while expression of SMI 311-ir in LCs appears only about a week later. At postnatal day 1 (P1) relatively few mGluR1-ir LCs were detected in the WM and at the border of WM and IGL. Later, their number increased sharply until P15 (6-7 fold) and decreased continuously between P15 and P135. SMI 311-ir LCs were not present at P1 and even at P8 only a few were observed in the WM or in infraganglionic positions. Their number increased gradually (12-14 fold) until adulthood when their number was stabilized at 8.000-10.000/cerebellum. At the same time the number of probably ectopic SMI 311-ir LCs decreased with age: at P22 about one third of them was found in "ectopic" position, whereas in the adult cat only about 10-12% of LCs's was either in the WM or scattered in the whole depth of the granular layer. These results suggest that: (1) most LCs appear in the cerebellar cortex postnatally; and (2) postnatal migration and incorporation of LCs to the cortex is a much longer process than previously expected, occurring even after the cytoarchitectonic built-up (about P65-P70 in cat) of

  2. First description of food-borne Salmonella enterica resistance regions R1 and R3 associated with IS26 elements

    Eduarda Gomes-Neves; Vera Manageiro; Eugénia Ferreira; José M Correia da Costa; Manuela Caniça


    In this study, we assessed the presence of IS26 in food-borne ASSuT-type Salmonella enterica isolates. A new genetic region (R3) wasdescribed, that included a C14 caspase gene between IS26 elements. R3 was present in two Salmonella Rissen isolates from a swine carcass anda meat handler, collected at the same abattoir. Furthermore, a new rearrangement of resistance region R1, harboring the blaTEM-1 gene flanked byIS26 elements, was identified in Salmonella Typhimurium and Salmonella 4,[5],12:i...

  3. First description of food-borne Salmonella enterica resistance regions R1 and R3 associated with IS26 elements

    Gomes-Neves, Eduarda; Manageiro, Vera; Ferreira, Eugénia; José M Correia da Costa; Caniça, Manuela


    In this study, we assessed the presence of IS26 in food-borne ASSuT-type Salmonella enterica isolates. A new genetic region (R3) was described, that included a C14 caspase gene between IS26 elements. R3 was present in two Salmonella Rissen isolates from a swine carcass and a meat handler, collected at the same abattoir. Furthermore, a new rearrangement of resistance region R1, harboring the blaTEM-1 gene flanked by IS26 elements, was identified in Salmonella Typhimurium and Salmonella 4,[5],1...

  4. Upgrading the electrical system of the IEA-R1 reactor to avoid triggering event of accidents

    Mello, Jose Roberto de; Madi Filho, Tufic, E-mail:, E-mail: [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    The IEA-R1 research reactor at the Institute of Energy and Nuclear Research (IPEN) is a research reactor open pool type, built and designed by the American firm 'Babcox and Wilcox', having as coolant and moderator demineralized light water and Beryllium and graphite, as reflectors. The power supply system is designed to meet the electricity demand required by the loads of the reactor (Security systems and systems not related to security) in different situations the plant can meet, such as during startup, normal operation at power, shutdown, maintenance, exchange of fuel elements and accident situations. Studies have been done on possible accident initiating events and deterministic techniques were applied to assess the consequences of such incidents. Thus, the methods used to identify and select the accident initiating events, the methods of analysis of accidents, including sequence of events, transient analysis and radiological consequences, have been described. Finally, acceptance criteria of radiological doses are described. Only a brief summary of the item concerning loss of electrical power will be presented. The loss of normal electrical power at the IEA-R1 reactor is very common. In the case of Electric External Power Loss, at the IEA-R1 reactor building, there may be different sequences of events, as described below. When the supply of external energy in the IEA-R1 facility fails, the Electrical Distribution Vital System, consisting of 4 (four) generators type 'UPS', starts operation, immediately and it will continue supplying power to the reactor control table, core cooling system and other security systems. To contribute to security, in the electric power failure, starts to operate the Emergency Cooling System (SRE). SRE has the function of removing residual heat from the core to prevent the melting of fuel elements in the event of loss of refrigerant to the core. Adding to the generators with batteries group system, new auxiliary

  5. CYP287A1 is a carotenoid 2-β-hydroxylase required for deinoxanthin biosynthesis in Deinococcus radiodurans R1.

    Zhou, Zhengfu; Zhang, Wei; Su, Shiyou; Chen, Ming; Lu, Wei; Lin, Min; Molnár, István; Xu, Yuquan


    The carotenoid deinoxanthin is a crucial resistance factor against various stresses in the radiation-resistant bacterium Deinococcus radiodurans. Disruption of the gene dr2473 encoding the cytochrome P450 CYP287A1 led to the accumulation of 2-deoxydeinoxanthin in D. radiodurans, demonstrating that CYP287A1 is a novel β-carotene 2-hydroxylase. The dr2473 knockout mutant was shown to be more sensitive to UV radiation and oxidative stress than the wild-type strain D. radiodurans R1, indicating that the C2 alcohol of deinoxanthin is important for antioxidant activity.

  6. ERK-GluR1 phosphorylation in trigeminal spinal subnucleus caudalis neurons is involved in pain associated with dry tongue

    Nakaya, Yuka; Tsuboi, Yoshiyuki; Okada-Ogawa, Akiko; Shinoda, Masamichi; Kubo, Asako; Chen, Jui Yen; Noma, Noboru; Batbold, Dulguun; Imamura, Yoshiki; Sessle, Barry J


    Background Dry mouth is known to cause severe pain in the intraoral structures, and many dry mouth patients have been suffering from intraoral pain. In development of an appropriate treatment, it is crucial to study the mechanisms underlying intraoral pain associated with dry mouth, yet the detailed mechanisms are not fully understood. To evaluate the mechanisms underlying pain related to dry mouth, the dry-tongue rat model was developed. Hence, the mechanical or heat nocifensive reflex, the phosphorylated extracellular signal-regulated kinase and phosphorylated GluR1-IR immunohistochemistries, and the single neuronal activity were examined in the trigeminal spinal subnucleus caudalis of dry-tongue rats. Results The head-withdrawal reflex threshold to mechanical, but not heat, stimulation of the tongue was significantly decreased on day 7 after tongue drying. The mechanical, but not heat, responses of trigeminal spinal subnucleus caudalis nociceptive neurons were significantly enhanced in dry-tongue rats compared to sham rats on day 7. The number of phosphorylated extracellular signal-regulated kinase-immunoreactive cells was also significantly increased in the trigeminal spinal subnucleus caudalis following noxious stimulation of the tongue in dry-tongue rats compared to sham rats on day 7. The decrement of the mechanical head-withdrawal reflex threshold (HWT) was reversed during intracisternal administration of the mitogen-activated protein kinase kinase 1 inhibitor, PD98059. The trigeminal spinal subnucleus caudalis neuronal activities and the number of phosphorylated extracellular signal-regulated kinase-immunoreactive cells following noxious mechanical stimulation of dried tongue were also significantly decreased following intracisternal administration of PD98059 compared to vehicle-administrated rats. Increased number of the phosphorylated GluR1-IR cells was observed in the trigeminal spinal subnucleus caudalis of dry-tongue rats, and the number of

  7. Common Variants in CYP2R1 and GC Genes Predict Vitamin D Concentrations in Healthy Danish Children and Adults

    Nissen, Ioanna; Rasmussen, Lone Banke; Ravn-Haren, Gitte


    Environmental factors such as diet, intake of vitamin D supplements and exposure to sunlight are known to influence serum vitamin D concentrations. Genetic epidemiology of vitamin D is in its infancy and a better understanding on how genetic variation influences vitamin D concentration is needed....... We aimed to analyse previously reported vitamin D-related polymorphisms in relation to serum 25(OH)D concentrations in 201 healthy Danish families with dependent children in late summer in Denmark. Serum 25(OH)D concentrations and a total of 25 SNPs in GC, VDR, CYP2R1, CYP24A1, CYP27B1, C10or88...

  8. The role of ptsP, orfT, and sss recombinase in root colonization by Pseudomonas flurescens Q8r1-96.

    Pseudomonas fluorescens Q8r1-96 produces 2,4-diacetylphloroglucinol (2,4-DAPG), a polyketide antibiotic that suppresses a wide variety of soilborne fungal pathogens, including Gaeumannomyces graminis var. tritici, which causes take-all disease of wheat. Strain Q8r1-96 is representative of the D-geno...

  9. Complete Genome Sequence of Clavibacter michiganensis subsp. insidiosus R1-1 Using PacBio Single-Molecule Real-Time Technology.

    Lu, You; Samac, Deborah A; Glazebrook, Jane; Ishimaru, Carol A


    We report here the complete genome sequence of Clavibacter michiganensis subsp. insidiosus R1-1, isolated in Minnesota, USA. The R1-1 genome, generated by a de novo assembly of PacBio sequencing data, is the first complete genome sequence available for this subspecies. Copyright © 2015 Lu et al.

  10. Synaptic function for the Nogo-66 receptor NgR1: regulation of dendritic spine morphology and activity-dependent synaptic strength.

    Lee, Hakjoo; Raiker, Stephen J; Venkatesh, Karthik; Geary, Rebecca; Robak, Laurie A; Zhang, Yu; Yeh, Hermes H; Shrager, Peter; Giger, Roman J


    In the mature nervous system, changes in synaptic strength correlate with changes in neuronal structure. Members of the Nogo-66 receptor family have been implicated in regulating neuronal morphology. Nogo-66 receptor 1 (NgR1) supports binding of the myelin inhibitors Nogo-A, MAG (myelin-associated glycoprotein), and OMgp (oligodendrocyte myelin glycoprotein), and is important for growth cone collapse in response to acutely presented inhibitors in vitro. After injury to the corticospinal tract, NgR1 limits axon collateral sprouting but is not important for blocking long-distance regenerative growth in vivo. Here, we report on a novel interaction between NgR1 and select members of the fibroblast growth factor (FGF) family. FGF1 and FGF2 bind directly and with high affinity to NgR1 but not to NgR2 or NgR3. In primary cortical neurons, ectopic NgR1 inhibits FGF2-elicited axonal branching. Loss of NgR1 results in altered spine morphologies along apical dendrites of hippocampal CA1 neurons in vivo. Analysis of synaptosomal fractions revealed that NgR1 is enriched synaptically in the hippocampus. Physiological studies at Schaffer collateral-CA1 synapses uncovered a synaptic function for NgR1. Loss of NgR1 leads to FGF2-dependent enhancement of long-term potentiation (LTP) without altering basal synaptic transmission or short-term plasticity. NgR1 and FGF receptor 1 (FGFR1) are colocalized to synapses, and mechanistic studies revealed that FGFR kinase activity is necessary for FGF2-elicited enhancement of hippocampal LTP in NgR1 mutants. In addition, loss of NgR1 attenuates long-term depression of synaptic transmission at Schaffer collateral-CA1 synapses. Together, our findings establish that physiological NgR1 signaling regulates activity-dependent synaptic strength and uncover neuronal NgR1 as a regulator of synaptic plasticity.

  11. Bioactive Benzofuran Derivatives from Cortex Mori Radicis, and Their Neuroprotective and Analgesic Activities Mediated by mGluR1

    Ya-Nan Wang


    Full Text Available Four new benzofuran-type stilbene glycosides and 14 known compounds including 8 benzofuran-type stilbenes and 6 flavonoids were isolated from the traditional Chinese medicine, Cortex Mori Radicis. The new compounds were identified as (9R-moracin P 3′-O-α-l-arabinopyranoside (1, (9R-moracin P 9-O-β-d-glucopyranoside (2, (9R-moracin P 3′-O-β-d-glucopyranoside (3, and (9R-moracin O 10-O-β-d-glucopyranoside (4 based on the spectroscopic interpretation and chemical analysis. Three benzofuran-type stilbenes, moracin O (5, R (7, and P (8 showed significant neuroprotective activity against glutamate-induced cell death in SK-N-SH cells. In addition, moracin O (5 and P (8 also demonstrated a remarkable inhibition of the acetic acid-induced pain. The molecular docking with metabotropic glutamate receptor 1 (mGluR1 results indicated that these neuroprotective benzofuran-type stilbenes might be the active analgesic components of the genus Morus, and acted by mediating the mGluR1 pathway.

  12. Structural insights into Ca(2+)-activated long-range allosteric channel gating of RyR1.

    Wei, Risheng; Wang, Xue; Zhang, Yan; Mukherjee, Saptarshi; Zhang, Lei; Chen, Qiang; Huang, Xinrui; Jing, Shan; Liu, Congcong; Li, Shuang; Wang, Guangyu; Xu, Yaofang; Zhu, Sujie; Williams, Alan J; Sun, Fei; Yin, Chang-Cheng


    Ryanodine receptors (RyRs) are a class of giant ion channels with molecular mass over 2.2 mega-Daltons. These channels mediate calcium signaling in a variety of cells. Since more than 80% of the RyR protein is folded into the cytoplasmic assembly and the remaining residues form the transmembrane domain, it has been hypothesized that the activation and regulation of RyR channels occur through an as yet uncharacterized long-range allosteric mechanism. Here we report the characterization of a Ca(2+)-activated open-state RyR1 structure by cryo-electron microscopy. The structure has an overall resolution of 4.9 Å and a resolution of 4.2 Å for the core region. In comparison with the previously determined apo/closed-state structure, we observed long-range allosteric gating of the channel upon Ca(2+) activation. In-depth structural analyses elucidated a novel channel-gating mechanism and a novel ion selectivity mechanism of RyR1. Our work not only provides structural insights into the molecular mechanisms of channel gating and regulation of RyRs, but also sheds light on structural basis for channel-gating and ion selectivity mechanisms for the six-transmembrane-helix cation channel family.

  13. Structure and magnetism in S r1 -xAxTc O3 perovskites: Importance of the A -site cation

    Reynolds, Emily; Avdeev, Maxim; Thorogood, Gordon J.; Poineau, Frederic; Czerwinski, Kenneth R.; Kimpton, Justin A.; Yu, Michelle; Kayser, Paula; Kennedy, Brendan J.


    The S r1 -xB axTc O3 (x =0 , 0.1, 0.2) oxides were prepared and their solid-state and magnetic structure studied as a function of temperature by x-ray and neutron powder diffraction. The refined Tc moments at room temperature and Néel temperatures for B a0.1S r0.9Tc O3 and B a0.2S r0.8Tc O3 were 2.32 (14 ) μβ and 2.11 (13 ) μβ and 714 ∘C and 702 ∘C , respectively. In contrast to expectations, the Néel temperature in the series S r1 -xAxTc O3 decreases with increasing Ba content. This observation is consistent with previous experimental measurements for the two series A M O3 (M =Ru , Mn; A =Ca , Sr, Ba) where the maximum magnetic ordering temperature was observed for A =Sr . Taken with these previous results the current work demonstrates the critical role of the A -site cation in the broadening of the π* bandwidth and ultimately the magnetic ordering temperature.

  14. Bacillus licheniformis BlaR1 L3 loop is a zinc metalloprotease activated by self-proteolysis.

    Stéphanie Berzigotti

    Full Text Available In Bacillus licheniformis 749/I, BlaP β-lactamase is induced by the presence of a β-lactam antibiotic outside the cell. The first step in the induction mechanism is the detection of the antibiotic by the membrane-bound penicillin receptor BlaR1 that is composed of two functional domains: a carboxy-terminal domain exposed outside the cell, which acts as a penicillin sensor, and an amino-terminal domain anchored to the cytoplasmic membrane, which works as a transducer-transmitter. The acylation of BlaR1 sensor domain by the antibiotic generates an intramolecular signal that leads to the activation of the L3 cytoplasmic loop of the transmitter by a single-point cleavage. The exact mechanism of L3 activation and the nature of the secondary cytoplasmic signal launched by the activated transmitter remain unknown. However, these two events seem to be linked to the presence of a HEXXH zinc binding motif of neutral zinc metallopeptidases. By different experimental approaches, we demonstrated that the L3 loop binds zinc ion, belongs to Gluzincin metallopeptidase superfamily and is activated by self-proteolysis.

  15. IL18 and IL18R1 polymorphisms, lung CT and fibrosis: a longitudinal study in coal miners

    Nadif, R.; Mintz, M.; Marzec, J.; Jedlicka, A.; Kauffmann, F.; Kleeberger, S.R. [INSERM, U780, Villejuif (France)


    It has been suggested that interleukin (IL)-18 plays a role in the development of inflammatory and fibrosing lung diseases. Associations of polymorphisms in the genes coding for IL-18 (108 /G-656T, C-607A, G-137C, T113G, C127T) and its receptor (IL8R1/C-69T) with coal workers' pneumoconiosis (CWP) were studied in 200 miners who were examined in 1990, 1994 and 1999. Coal-dust exposure was assessed according to job history and ambient measures. The main health outcome was lung computed tomography (CT) score in 1990. Internal coherence was assessed by studying CT score in 1994, 4-yr change in CT score and CWP incidence and prevalence. CT score in 1990 was a good predictor of radiographic grade in 1999 and, therefore, an appropriate subclinical quantitative trait. The IL18 -137C allele was associated with lower CT score in 1990 and 1994 (11.24 versus 1.69 and 1.57 versus 2.46, respectively), slower progression of CT score between 1990 and 1994 and lower pneumoconiosis prevalence in 1999 relative to the G allele (0.33 versus 0.77 and 8.2 versus 19.6%, respectively). Smoking- or dust-adjustment, and stratification on IL18R1 genotype and adjustment for haplotype effects did not change the conclusions. In conclusion, the results of the present study suggest a role for IL18 in reducing the development of this fibrosing lung disease.

  16. Experimental study of the temperature distribution in the TRIGA IPR-R1 Brazilian research reactor; Investigacao experimental da distribuicao de temperaturas no reator nuclear de pesquisa TRIGA IPR-R1

    Mesquita, Amir Zacarias


    The TRIGA-IPR-R1 Research Nuclear Reactor has completed 44 years in operation in November 2004. Its initial nominal thermal power was 30 kW. In 1979 its power was increased to 100 kW by adding new fuel elements to the reactor. Recently some more fuel elements were added to the core increasing the power to 250 kW. The TRIGA-IPR-R1 is a pool type reactor with a natural circulation core cooling system. Although the large number of experiments had been carried out with this reactor, mainly on neutron activation analysis, there is not many data on its thermal-hydraulics processes, whether experimental or theoretical. So a number of experiments were carried out with the measurement of the temperature inside the fuel element, in the reactor core and along the reactor pool. During these experiments the reactor was set in many different power levels. These experiments are part of the CDTN/CNEN research program, and have the main objective of commissioning the TRIGA-IPR-R1 reactor for routine operation at 250 kW. This work presents the experimental and theoretical analyses to determine the temperature distribution in the reactor. A methodology for the calibration and monitoring the reactor thermal power was also developed. This methodology allowed adding others power measuring channels to the reactor by using thermal processes. The fuel thermal conductivity and the heat transfer coefficient from the cladding to the coolant were also experimentally valued. lt was also presented a correlation for the gap conductance between the fuel and the cladding. The experimental results were compared with theoretical calculations and with data obtained from technical literature. A data acquisition and processing system and a software were developed to help the investigation. This system allows on line monitoring and registration of the main reactor operational parameters. The experiments have given better comprehension of the reactor thermal-fluid dynamics and helped to develop numerical

  17. mGluR1-mediated excitation of cerebellar GABAergic interneurons requires both G protein-dependent and Src-ERK1/2-dependent signaling pathways.

    Hideo Kubota

    Full Text Available Stimulation of type I metabotropic glutamate receptors (mGluR1/5 in several neuronal types induces slow excitatory responses through activation of transient receptor potential canonical (TRPC channels. GABAergic cerebellar molecular layer interneurons (MLIs modulate firing patterns of Purkinje cells (PCs, which play a key role in cerebellar information processing. MLIs express mGluR1, and activation of mGluR1 induces an inward current, but its precise intracellular signaling pathways are unknown. We found that mGluR1 activation facilitated spontaneous firing of mouse cerebellar MLIs through an inward current mediated by TRPC1 channels. This mGluR1-mediated inward current depends on both G protein-dependent and -independent pathways. The nonselective protein tyrosine kinase inhibitors genistein and AG490 as well as the selective extracellular signal-regulated kinase 1/2 (ERK1/2 inhibitors PD98059 and SL327 suppressed the mGluR1-mediated current responses. Following G protein blockade, the residual mGluR1-mediated inward current was significantly reduced by the selective Src tyrosine kinase inhibitor PP2. In contrast to cerebellar PCs, GABAB receptor activation in MLIs did not alter the mGluR1-mediated inward current, suggesting that there is no cross-talk between mGluR1 and GABAB receptors in MLIs. Thus, activation of mGluR1 facilitates firing of MLIs through the TRPC1-mediated inward current, which depends on not only G protein-dependent but also Src-ERK1/2-dependent signaling pathways, and consequently depresses the excitability of cerebellar PCs.

  18. Transgenic wheat expressing Thinopyrum intermedium MYB transcription factor TiMYB2R-1 shows enhanced resistance to the take-all disease.

    Liu, Xin; Yang, Lihua; Zhou, Xianyao; Zhou, Miaoping; Lu, Yan; Ma, Lingjian; Ma, Hongxiang; Zhang, Zengyan


    The disease take-all, caused by the fungus Gaeumannomyces graminis, is one of the most destructive root diseases of wheat worldwide. Breeding resistant cultivars is an effective way to protect wheat from take-all. However, little progress has been made in improving the disease resistance level in commercial wheat cultivars. MYB transcription factors play important roles in plant responses to environmental stresses. In this study, an R2R3-MYB gene in Thinopyrum intermedium, TiMYB2R-1, was cloned and characterized. The gene sequence includes two exons and an intron. The expression of TiMYB2R-1 was significantly induced following G. graminis infection. An in vitro DNA binding assay proved that TiMYB2R-1 protein could bind to the MYB-binding site cis-element ACI. Subcellular localization assays revealed that TiMYB2R-1 was localized in the nucleus. TiMYB2R-1 transgenic wheat plants were generated, characterized molecularly, and evaluated for take-all resistance. PCR and Southern blot analyses confirmed that TiMYB2R-1 was integrated into the genomes of three independent transgenic wheat lines by distinct patterns and the transgene was heritable. Reverse transcription-PCR and western blot analyses revealed that TiMYB2R-1 was highly expressed in the transgenic wheat lines. Based on disease response assessments for three successive generations, the significantly enhanced resistance to take-all was observed in the three TiMYB2R-1-overexpressing transgenic wheat lines. Furthermore, the transcript levels of at least six wheat defence-related genes were significantly elevated in the TiMYB2R-1 transgenic wheat lines. These results suggest that engineering and overexpression of TiMYB2R-1 may be used for improving take-all resistance of wheat and other cereal crops.

  19. The stress-related, rhizobial small RNA RcsR1 destabilizes the autoinducer synthase encoding mRNA sinI in Sinorhizobium meliloti.

    Baumgardt, Kathrin; Šmídová, Klára; Rahn, Helen; Lochnit, Günter; Robledo, Marta; Evguenieva-Hackenberg, Elena


    Quorum sensing is a cell density-dependent communication system of bacteria relying on autoinducer molecules. During the analysis of the post-transcriptional regulation of quorum sensing in the nitrogen fixing plant symbiont Sinorhizobium meliloti, we predicted and verified a direct interaction between the 5'-UTR of sinI mRNA encoding the autoinducer synthase and a small RNA (sRNA), which we named RcsR1. In vitro, RcsR1 prevented cleavage in the 5'-UTR of sinI by RNase E and impaired sinI translation. In line with low ribosomal occupancy and transcript destabilization upon binding of RcsR1 to sinI, overproduction of RcsR1 in S. meliloti resulted in lower level and shorter half-life of sinI mRNA, and in decreased autoinducer amount. Although RcsR1 can influence quorum sensing via sinI, its level did not vary at different cell densities, but decreased under salt stress and increased at low temperature. We found that RcsR1 and its stress-related expression pattern, but not the interaction with sinI homologs, are conserved in Sinorhizobium, Rhizobium and Agrobacterium. Consistently, overproduction of RcsR1 in S. meliloti and Agrobacterium tumefaciens inhibited growth at high salinity. We identified conserved targets of RcsR1 and showed that most conserved interactions and the effect on growth under salt stress are mediated by the first stem-loop of RcsR1, while its central part is responsible for the species-specific interaction with sinI. We conclude that RcsR1 is an ancient, stress-related riboregulator in rhizobia and propose that it links stress responses to quorum sensing in S. meliloti.

  20. Development of methodology for characterization of cartridge filters from the IEA-R1 using the Monte Carlo method; Desenvolvimento de uma metodologia para caracterizacao do filtro cuno do reator IEA-R1 utilizando o Metodo Monte Carlo

    Costa, Priscila


    The Cuno filter is part of the water processing circuit of the IEA-R1 reactor and, when saturated, it is replaced and becomes a radioactive waste, which must be managed. In this work, the primary characterization of the Cuno filter of the IEA-R1 nuclear reactor at IPEN was carried out using gamma spectrometry associated with the Monte Carlo method. The gamma spectrometry was performed using a hyperpure germanium detector (HPGe). The germanium crystal represents the detection active volume of the HPGe detector, which has a region called dead layer or inactive layer. It has been reported in the literature a difference between the theoretical and experimental values when obtaining the efficiency curve of these detectors. In this study we used the MCNP-4C code to obtain the detector calibration efficiency for the geometry of the Cuno filter, and the influence of the dead layer and the effect of sum in cascade at the HPGe detector were studied. The correction of the dead layer values were made by varying the thickness and the radius of the germanium crystal. The detector has 75.83 cm{sup 3} of active volume of detection, according to information provided by the manufacturer. Nevertheless, the results showed that the actual value of active volume is less than the one specified, where the dead layer represents 16% of the total volume of the crystal. A Cuno filter analysis by gamma spectrometry has enabled identifying energy peaks. Using these peaks, three radionuclides were identified in the filter: {sup 108m}Ag, {sup 110m}Ag and {sup 60}Co. From the calibration efficiency obtained by the Monte Carlo method, the value of activity estimated for these radionuclides is in the order of MBq. (author)

  1. A divergent asymmetric approach to aza-spiropyran derivative and (1S,8aR-1-hydroxyindolizidine

    Huang Pei-Qiang


    Full Text Available Abstract Background Spiroketals and the corresponding aza-spiroketals are the structural features found in a number of bioactive natural products, and in compounds possessing photochromic properties for use in the area of photochemical erasable memory, self-development photography, actinometry, displays, filters, lenses of variable optical density, and photomechanical biomaterials etc. And (1R,8aS-1-hydroxyindolizidine (3 has been postulated to be a biosynthetic precursor of hydroxylated indolizidines such as (+-lentiginosine 1, (--2-epilentiginosine 2 and (--swainsonine, which are potentially useful antimetastasis drugs for the treatment of cancer. In continuation of a project aimed at the development of enantiomeric malimide-based synthetic methodology, we now report a divergent, concise and highly diastereoselective approach for the asymmetric syntheses of an aza-spiropyran derivative 7 and (1S,8aR-1-hydroxyindolizidine (ent-3. Results The synthesis of aza-spiropyran 7 started from the Grignard addition of malimide 4. Treatment of the THP-protected 4-hydroxybutyl magnesium bromide with malimide 4 at -20°C afforded N,O-acetal 5a as an epimeric mixture in a combined yield of 89%. Subjection of the diastereomeric mixture of N,O-acetal 5a to acidic conditions for 0.5 h resulted in the formation of the desired functionalized aza-spiropyran 7 as a single diastereomer in quantitative yield. The stereochemistry of the aza-spiropyran 7 was determined by NOESY experiment. For the synthesis of ent-3, aza-spiropyran 7, or more conveniently, N,O-acetal 5a, was converted to lactam 6a under standard reductive dehydroxylation conditions in 78% or 77% yield. Reduction of lactam 6a with borane-dimethylsulfide provided pyrrolidine 8 in 95% yield. Compound 8 was then converted to 1-hydroxyindolizidine ent-3 via a four-step procedure, namely, N-debenzylation/O-mesylation/Boc-cleavage/cyclization, and O-debenzylation. Alternatively, amino alcohol 8 was mesylated

  2. Spin and orbital disordering by hole doping in P r1 -xC axV O3

    Reehuis, M.; Ulrich, C.; Abdala, P. M.; Pattison, P.; Khaliullin, G.; Fujioka, J.; Miyasaka, S.; Tokura, Y.; Keimer, B.


    High-resolution powder x-ray diffraction and single-crystal neutron diffraction were used to investigate the crystal structure and magnetic ordering of the compound P r1 -xC axV O3 (0 ≤x ≤0.3 ), which undergoes an insulator-to-metal transition for x ˜0.23 . Since the ionic radii of P r3 + and C a2 + are almost identical and structural disorder is minimal, P r1 -xC axV O3 is a good model system for the influence of hole doping on the spin and orbital correlations in transition metal oxides. The end member PrV O3 is a Mott-Hubbard insulator, which exhibits a structural phase transition at TS=180 K from an orthorhombic to a monoclinic structure with space groups Pbnm and P 21/b , respectively. This transition is associated with the onset of orbital ordering and strong Jahn-Teller distortions of the V O6 octahedra. Antiferromagnetic C -type order with vanadium moments oriented in the a b plane is observed below TN=140 K . Upon cooling, the vanadium moments induce a progressive magnetic polarization of the praseodymium sublattice, resulting in a ferrimagnetic structure with coexisting modes (Cx, Fy) and (Fx, Cy). In the insulating range of the P r1 -xC axV O3 phase diagram, Ca doping reduces both the orbital and magnetic transition temperatures so that TS=108 K and TN=95 K for x =0.20 . The Jahn-Teller distortions and ordered vanadium moments also decrease upon doping. In a metallic sample with x =0.30 , Jahn-Teller distortions and long-range orbital ordering are no longer observable, and the average crystal structure remains orthorhombic down to low temperature. However, broadening of some lattice Bragg reflections indicate a significant increase in lattice strain. Antiferromagnetic short-range order with a weak ordered moment of 0.14(3) μB per vanadium atom could still be observed on the vanadium site below T ˜60 K . We discuss these observations in terms of doping-induced spin-orbital polaron formation.

  3. TRAIL receptor gene editing unveils TRAIL-R1 as a master player of apoptosis induced by TRAIL and ER stress.

    Dufour, Florent; Rattier, Thibault; Constantinescu, Andrei Alexandru; Zischler, Luciana; Morlé, Aymeric; Ben Mabrouk, Hazem; Humblin, Etienne; Jacquemin, Guillaume; Szegezdi, Eva; Delacote, Fabien; Marrakchi, Naziha; Guichard, Gilles; Pellat-Deceunynck, Catherine; Vacher, Pierre; Legembre, Patrick; Garrido, Carmen; Micheau, Olivier


    TRAIL induces selective tumor cell death through TRAIL-R1 and TRAIL-R2. Despite the fact that these receptors share high structural homologies, induction of apoptosis upon ER stress, cell autonomous motility and invasion have solely been described to occur through TRAIL-R2. Using the TALEN gene-editing approach, we show that TRAIL-R1 can also induce apoptosis during unresolved unfolded protein response (UPR). Likewise, TRAIL-R1 was found to co-immunoprecipitate with FADD and caspase-8 during ER stress. Its deficiency conferred resistance to apoptosis induced by thaspigargin, tunicamycin or brefeldin A. Our data also demonstrate that tumor cell motility and invasion-induced by TRAIL-R2 is not cell autonomous but induced in a TRAIL-dependant manner. TRAIL-R1, on the other hand, is unable to trigger cell migration owing to its inability to induce an increase in calcium flux. Importantly, all the isogenic cell lines generated in this study revealed that apoptosis induced TRAIL is preferentially induced by TRAIL-R1. Taken together, our results provide novel insights into the physiological functions of TRAIL-R1 and TRAIL-R2 and suggest that targeting TRAIL-R1 for anticancer therapy is likely to be more appropriate owing to its lack of pro-motile signaling capability.

  4. The impact of R1and R3a genes on tuber resistance to late blight of the potato breeding clones

    Zoteyeva Nadezhda


    Full Text Available Potato breeding clones were evaluated for resistance to late blight (agent Phytophthora infestans using tuber inoculation tests and for presence of the resistance alleles of R1 and R3a genes in polymerase chain reaction tests. Among clones tested those expressing high, moderate and low resistance were identified. The data were analysed for the impact of R1 and R3a genes on tuber resistance to late blight in tested plant material. In previous evaluations performed on smaller amount of clones the tuber resistance levels significantly depended on presence/absence of the resistance allele of R3a gene and did not depend on presence of R1 gene allele. In the current study the statistical analyses did not prove the significant difference in resistance levels depending on presence of the resistance alleles, neither of R1 gene, nor of R3a gene. Tuber resistant clones bearing R3a gene resistance alleles still noticeably prevailed over the clones bearing the alleles of R1 gene as well as over the clones bearing the no resistance alleles of both genes. In several cases the resistance of clones with detected resistance allele of R1 gene was higher compared to those derived from the same crosses and showing amplification of the allele of R3a gene or those with no resistance alleles. Clones accumulating the resistance alleles of both (R1 and R3a genes expressed high tuber resistance accompanied by necrotic reaction.

  5. Decoy Receptor DcR1 Is Induced in a p50/Bcl3-Dependent Manner and Attenuates the Efficacy of Temozolomide.

    Mansour, Nassir M; Bernal, Giovanna M; Wu, Longtao; Crawley, Clayton D; Cahill, Kirk E; Voce, David J; Balyasnikova, Irina V; Zhang, Wei; Spretz, Ruben; Nunez, Luis; Larsen, Gustavo F; Weichselbaum, Ralph R; Yamini, Bakhtiar


    Temozolomide is used widely to treat malignant glioma, but the overall response to this agent is generally poor. Resistance to DNA-damaging drugs such as temozolomide has been related to the induction of antiapoptotic proteins. Specifically, the transcription factor NF-κB has been suggested to participate in promoting the survival of cells exposed to chemotherapy. To identify factors that modulate cytotoxicity in the setting of DNA damage, we used an unbiased strategy to examine the NF-κB-dependent expression profile induced by temozolomide. By this route, we defined the decoy receptor DcR1 as a temozolomide response gene induced by a mechanism relying upon p50/NF-κB1. A conserved NF-κB-binding sequence (κB-site) was identified in the proximal promoter and was demonstrated to be required for DcR1 induction by temozolomide. Loss-of-function and gain-of-function studies reveal that the atypical IκB protein, Bcl3, is also required for induction of DcR1 by temozolomide. Mechanistically, DcR1 attenuates temozolomide efficacy by blunting activation of the Fas receptor pathway in p53(+/+) glioma cells. Intracranial xenograft studies show that DcR1 depletion in glioma cells enhances the efficacy of temozolomide. Taken together, our results show how DcR1 upregulation mediates temozolomide resistance and provide a rationale for DcR1 targeting as a strategy to sensitize gliomas to this widely used chemotherapy.

  6. The heat-shock DnaK protein is required for plasmid R1 replication and it is dispensable for plasmid ColE1 replication.

    Giraldo-Suárez, R; Fernández-Tresguerres, E; Díaz-Orejas, R; Malki, A; Kohiyama, M


    Plasmid R1 replication in vitro is inactive in extracts prepared from a dnaK756 strain but is restored to normal levels upon addition of purified DnaK protein. Replication of R1 in extracts of a dnaKwt strain can be specifically inhibited with polyclonal antibodies against DnaK. RepA-dependent replication of R1 in dnaK756 extracts supplemented with DnaKwt protein at maximum concentration is partially inhibited by rifampicin and it is severely inhibited at sub-optimal concentrations of DnaK protein. The copy number of a run-away R1 vector is reduced in a dnaK756 background at 30 degrees C and at 42 degrees C the amplification of the run-away R1 vector is prevented. However a runaway R1 vector containing dnaK gene allows the amplification of the plasmid at high temperature. These data indicate that DnaK is required for both in vitro and in vivo replication of plasmid R1 and show a partial compensation for the low level of DnaK by RNA polymerase. In contrast ColE1 replication is not affected by DnaK as indicated by the fact that ColE1 replicates with the same efficiency in extracts from dnaKwt and dnaK756 strains. Images PMID:8265367

  7. SIGN-R1 and complement factors are involved in the systemic clearance of radiation-induced apoptotic cells in whole-body irradiated mice

    Park, Jin-Yeon; Loh, SoHee; Cho, Eun-hee [Department of Biomedical Science & Technology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701 (Korea, Republic of); Choi, Hyeong-Jwa [Division of Radiation Effect, Korea Institute of Radiological and Medical Sciences, 215-4, 75 Nowon gil Nowon-Gu, Seoul, 139-706 (Korea, Republic of); Na, Tae-Young [College of Pharmacy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-741 (Korea, Republic of); Nemeno, Judee Grace E.; Lee, Jeong Ik [Regenerative Medicine Laboratory, Department of Veterinary Medicine, College of Veterinary Medicine, Konkuk University, Seoul, 143-701 (Korea, Republic of); Yoon, Taek Joon [Department of Food and Nutrition, Yuhan College, Bucheon, Gyeonggi-do, 422-749 (Korea, Republic of); Choi, In-Soo [Department of Infectious Diseases, College of Veterinary Medicine, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701 (Korea, Republic of); Lee, Minyoung [Division of Radiation Effect, Korea Institute of Radiological and Medical Sciences, 215-4, 75 Nowon gil Nowon-Gu, Seoul, 139-706 (Korea, Republic of); Lee, Jae-Seon [Department of Biomedical Sciences, College of Medicine, Inha University, Incheon, 400-712 (Korea, Republic of); Kang, Young-Sun, E-mail: [Department of Biomedical Science & Technology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701 (Korea, Republic of); Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701 (Korea, Republic of)


    Although SIGN-R1-mediated complement activation pathway has been shown to enhance the systemic clearance of apoptotic cells, the role of SIGN-R1 in the clearance of radiation-induced apoptotic cells has not been characterized and was investigated in this study. Our data indicated that whole-body γ-irradiation of mice increased caspase-3{sup +} apoptotic lymphocyte numbers in secondary lymphoid organs. Following γ-irradiation, SIGN-R1 and complements (C4 and C3) were simultaneously increased only in the mice spleen tissue among the assessed tissues. In particular, C3 was exclusively activated in the spleen. The delayed clearance of apoptotic cells was markedly prevalent in the spleen and liver of SIGN-R1 KO mice, followed by a significant increase of CD11b{sup +} cells. These results indicate that SIGN-R1 and complement factors play an important role in the systemic clearance of radiation-induced apoptotic innate immune cells to maintain tissue homeostasis after γ-irradiation. - Highlights: • Splenic SIGN-R1{sup +} macrophages are activated after γ-irradiation. • C3 and C4 levels increased and C3 was activated in the spleen after γ-irradiation. • SIGN-R1 mediated the systemic clearance of radiation-induced apoptotic cells in spleen and liver.

  8. International benchmark study of advanced thermal hydraulic safety analysis codes against measurements on IEA-R1 research reactor

    Hainoun, A., E-mail: [Atomic Energy Commission of Syria (AECS), Nuclear Engineering Department, P.O. Box 6091, Damascus (Syrian Arab Republic); Doval, A. [Nuclear Engineering Department, Av. Cmdt. Luis Piedrabuena 4950, C.P. 8400 S.C de Bariloche, Rio Negro (Argentina); Umbehaun, P. [Centro de Engenharia Nuclear – CEN, IPEN-CNEN/SP, Av. Lineu Prestes 2242-Cidade Universitaria, CEP-05508-000 São Paulo, SP (Brazil); Chatzidakis, S. [School of Nuclear Engineering, Purdue University, West Lafayette, IN 47907 (United States); Ghazi, N. [Atomic Energy Commission of Syria (AECS), Nuclear Engineering Department, P.O. Box 6091, Damascus (Syrian Arab Republic); Park, S. [Research Reactor Design and Engineering Division, Basic Science Project Operation Dept., Korea Atomic Energy Research Institute (Korea, Republic of); Mladin, M. [Institute for Nuclear Research, Campului Street No. 1, P.O. Box 78, 115400 Mioveni, Arges (Romania); Shokr, A. [Division of Nuclear Installation Safety, Research Reactor Safety Section, International Atomic Energy Agency, A-1400 Vienna (Austria)


    Highlights: • A set of advanced system thermal hydraulic codes are benchmarked against IFA of IEA-R1. • Comparative safety analysis of IEA-R1 reactor during LOFA by 7 working teams. • This work covers both experimental and calculation effort and presents new out findings on TH of RR that have not been reported before. • LOFA results discrepancies from 7% to 20% for coolant and peak clad temperatures are predicted conservatively. - Abstract: In the framework of the IAEA Coordination Research Project on “Innovative methods in research reactor analysis: Benchmark against experimental data on neutronics and thermal hydraulic computational methods and tools for operation and safety analysis of research reactors” the Brazilian research reactor IEA-R1 has been selected as reference facility to perform benchmark calculations for a set of thermal hydraulic codes being widely used by international teams in the field of research reactor (RR) deterministic safety analysis. The goal of the conducted benchmark is to demonstrate the application of innovative reactor analysis tools in the research reactor community, validation of the applied codes and application of the validated codes to perform comprehensive safety analysis of RR. The IEA-R1 is equipped with an Instrumented Fuel Assembly (IFA) which provided measurements for normal operation and loss of flow transient. The measurements comprised coolant and cladding temperatures, reactor power and flow rate. Temperatures are measured at three different radial and axial positions of IFA summing up to 12 measuring points in addition to the coolant inlet and outlet temperatures. The considered benchmark deals with the loss of reactor flow and the subsequent flow reversal from downward forced to upward natural circulation and presents therefore relevant phenomena for the RR safety analysis. The benchmark calculations were performed independently by the participating teams using different thermal hydraulic and safety

  9. Crystal structure of 2-{(R-[1-(4-bromophenylethyl]iminomethyl}-4-(phenyldiazenylphenol, a chiral photochromic Schiff base

    Ryoji Moriwaki


    Full Text Available The title chiral photochromic Schiff base compound, C21H18BrN3O, was synthesized from (R-(+-1-(4-bromophenylethylamine and the salicylaldehyde of an azobenzene derivative. The molecule corresponds to the phenol–imine tautomer, the C=N and N—C bond distances being 1.285 (3 and 1.470 (3 Å, respectively. The diazenyl group adopts a trans form, with an N=N distance of 1.256 (3 Å. The hydroxy group is involved in intramolecular O—H...N hydrogen bonding. In the crystal, C—H...π interactions consolidate the crystal packing of one-dimensional chains, which exhibits short intermolecular Br...C contacts of 3.400 (3 Å.

  10. Transient cases analyses of the TRIGA IPR-R1 using thermal hydraulic and neutron kinetic coupled codes

    Reis, Patricia A.L.; Costa, Antonella L.; Pereira, Claubia; Veloso, Maria A.F.; Scari, Maria E., E-mail:, E-mail:, E-mail:, E-mail:, E-mail: [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Departamento de Engenharia Nuclear; Instituto Nacional de Ciencias e Tecnologia de Reatores Nucleares Inovadores (INCT/CNPq), Belo Horizonte (Brazil); Miro, Rafael; Verdu, Gumersindo, E-mail:, E-mail: [Universidad Politecnica de Valencia (Spain). Departamento de Ingenieria Quimica y Nuclear


    Simulations and analyses of nuclear reactors have been improved by utilization of coupled thermal-hydraulic (TH) and neutron kinetics (NK) system codes especially to simulate transients that involve strong feedback effects between NK and TH. The TH-NK coupling technique was initially developed and used to simulate the behavior of power reactors; however, several coupling methodologies are now being applied for research reactors. This work presents the coupling methodology application between RELAP5 and PARCS codes using as a model the TRIGA IPR-R1 research reactor. Analyses of steady state and transient conditions and comparisons with results from simulations using only the RELAP5 code are being presented in this paper. (author)

  11. Roles of the canonical myomiRs mi R-1,-133 and-206 in cell development and disease

    Keith; Richard; Mitchelson; Wen-Yan; Qin


    Micro RNAs are small non-coding RNAs that participate in different biological processes, providing subtle combinational regulation of cellular pathways, often by regulating components of signalling pathways. Aberrant expression of mi RNAs is an important factor in the development and progression of disease. The canonical myomi Rs(mi R-1,-133 and-206) are central to the development and health of mammalian skeletal and cardiac muscles, but new findings show they have regulatory roles in the development of other mammalian non-muscle tissues, including nerve, brain structures, adipose and some specialised immunological cells. Moreover, the deregulation of myomi R expression is associated with a variety of different cancers, where typically they have tumor suppressor functions, although examples of an oncogenic role illustrate their diverse function in different cell environments. This review examines the involvement of the related myomi Rs at the crossroads between cell development/tissue regeneration/tissue inflammation responses, and cancer development.

  12. Invisible watermarking optical camera communication and compatibility issues of IEEE 802.15.7r1 specification

    Le, Nam-Tuan


    Copyright protection and information security are two most considered issues of digital data following the development of internet and computer network. As an important solution for protection, watermarking technology has become one of the challenged roles in industry and academic research. The watermarking technology can be classified by two categories: visible watermarking and invisible watermarking. With invisible technique, there is an advantage on user interaction because of the visibility. By applying watermarking for communication, it will be a challenge and a new direction for communication technology. In this paper we will propose one new research on communication technology using optical camera communications (OCC) based invisible watermarking. Beside the analysis on performance of proposed system, we also suggest the frame structure of PHY and MAC layer for IEEE 802.15.7r1 specification which is a revision of visible light communication (VLC) standardization.

  13. A simple approach to solving the kinematics of the 4-UPS/PS (3R1T) parallel manipulator

    Gallardo-Alvarado, Jaime; Gracio-Murillo, Mario A. [Instituto Tecnologico de Celaya, Celaya (Mexico); Islam, Md. Nazrul [Universiti Malasya Sabah, Sabah (Malaysia); Abedinnasab, Mohammad H. [Rowan University, New Jersey (United States)


    This work reports on the position, velocity and acceleration analyses of a four-degrees-of-freedom parallel manipulator, 4-DoF-PM for brevity, which generates Three-rotation-one-translation (3R1T) motion. Nearly closed-form solutions to solve the forward displacement analysis are easily obtained based on closure equations formulated upon linear combinations of the coordinates of three non-collinear points embedded in the moving platform. Then, the input-output equations of velocity and acceleration of the robot manipulator are systematically established by resorting to the theory of screws. To this end, the Klein form of the Lie algebra se(3) of the Euclidean group SE(3) is systematically applied to the velocity and reduced acceleration state in screw form of the moving platform cancelling the passive joint rates of the parallel manipulator. Numerical examples, which are confirmed by means of commercially available software, are provided to show the application of the method.

  14. Phase gap in pseudoternary R1-yRy'Mn2X2-xXx' compounds

    Wang, J. L.; Kennedy, S. J.; Campbell, S. J.; Hofmann, M.; Dou, S. X.


    Our neutron diffraction investigation of PrMn2Ge2-xSix reveals a clear separation into two magnetic phases, canted ferromagnetic (Fmc) and antiferromagnetic (AFmc), between x = 1.0 and 1.2 and a commensurate phase gap in the lattice, due to magnetostrictive distortion. This remarkable magnetoelastic phenomenon is driven by a nonuniform atomic distribution on the X site which in turn produces subtle variations in the local lattice and abrupt changes in the Mn-Mn magnetic exchange interaction. Our results show that coexistence of Fmc and AFmc phases depends on lattice parameter, chemical pressure from the rare-earth and metalloid sites, and local lattice strain distributions. We demonstrate that these magnetostructural correlations act across the entire family of R1-yRy'Mn2X2-xXx' compounds.

  15. [3H]Azidodantrolene photoaffinity labeling, synthetic domain peptides and monoclonal antibody reactivity identify the dantrolene binding sequence on RyR1

    Paul-Pletzer, Kalanethee; Yamamoto, Takeshi; Bhat, Manju B.; Ma, Jianjie; Ikemoto, Noriaki; Jimenez, Leslie S.; Morimoto, Hiromi; Williams, Philip G.; Parness, Jerome


    Dantrolene is a drug that suppresses intracellular Ca2+ release from sarcoplasmic reticulum in normal skeletal muscle and is used as a therapeutic agent in individuals susceptible to malignant hyperthermia. Though its precise mechanism of action has not been elucidated, we have identified the N-terminal region (amino acids 1-1400) of the skeletal muscle isoform of the ryanodine receptor (RyR1), the primary Ca2+ release channel in sarcoplasmic reticulum, as a molecular target for dantrolene using the photoaffinity analog [3H]azidodantrolene(1). Here, we demonstrate that heterologously expressed RyR1 retains its capacity to be specifically labeled with [3H]azidodantrolene,indicating that muscle specific factors are not required for this ligand-receptor interaction. Synthetic domain peptides of RyR1, previously shown to affect RyR1 function in vitro and in vivo, were exploited as potential drug binding site mimics and used in photoaffinity labeling experiments. Only DP1 and DP1-2, peptide s containing the amino acid sequence corresponding to RyR1 residues 590-609, were specifically labeled by [3H]azidodantrolene. A monoclonal anti-RyR1 antibody which recognizes RyR1 and its 1400 amino acid N-terminal fragment, recognizes DP1 and DP1-2 in both Western blots and immunoprecipitation assays, and specifically inhibits [3H]azidodantrolene photolabeling of RyR1 and its N-terminal fragment in sarcoplasmic reticulum. Our results indicate that synthetic domain peptides can mimic a native, ligand binding conformation in vitro, and that the dantrolene binding site and the epitope for the monoclonal antibody on RyR1 are equivalent and composed of amino-acids 590-609.

  16. Characterization of beta-R1, a gene that is selectively induced by interferon beta (IFN-beta) compared with IFN-alpha.

    Rani, M R; Foster, G R; Leung, S; Leaman, D; Stark, G R; Ransohoff, R M


    We report preliminary characterization of a gene designated beta-R1, which is selectively expressed in response to interferon beta (IFN-beta) compared with IFN-alpha. In human astrocytoma cells, beta-R1 was induced to an equivalent extent by 10 IU/mL IFN-beta or 2500 IU/mL IFN-alpha2. To address the mechanism of this differential response, we analyzed induction of the beta-R1 gene in fibrosarcoma cells and derivative mutant cells lacking components required for signaling by type I IFNs. beta-R1 was readily induced by IFN-beta in the parental 2fTGH cell line, but not by recombinant IFN-alpha2, IFN-alpha Con1, or a mixture of IFN-alpha subtypes. IFN-alpha8 induced beta-R1 weakly. beta-R1 was not induced by IFN-beta in mutant cell lines U2A, U3A, U4A, and U6A, which lack, respectively, p48, STAT1, JAK1, and STAT2. U5A cells, which lack the Ifnar 2.2 component of the IFN-alpha and -beta receptor, also failed to express beta-R1. U1A cells are partially responsive to IFN-beta and IFN-alpha8 but lacked beta-R1 expression, indicating that TYK2 protein is essential for induction of this gene. Taken together, these results suggest that the expression of beta-R1 in response to type I IFN requires IFN-stimulated gene factor 3 plus an additional component, which is more efficiently formed on induction by IFN-beta compared with IFN-alpha.

  17. Effects of PPP1R1B (DARPP-32 Polymorphism on Feedback-related Brain Potentials across the Life Span

    Dorothea eHämmerer


    Full Text Available Maximizing gains during probabilistic reinforcement learning requires the updating of choice–outcome expectations at the time when the feedback about a specific choice or action is given. Extant theories and evidence suggest that dopaminergic modulation plays a crucial role in reinforcement learning and the updating of choice–outcome expectations. Furthermore, recently a positive component of the event-related potential (ERP about 200 msec (P2 after feedback has been suggested to reflect such updating. The efficacy of dopaminergic modulation changes across the life span. However, to date investigations of age-related differences in feedback-related P2 during reinforcement learning are still scarce. The present study thus aims to investigate whether individual differences in the feedback-related P2 would be associated with polymorphic variations in a dopamine relevant gene PPP1R1B (also known as DARPP-32 and whether the genetic effect may differ between age groups. We observed larger P2 amplitudes in individuals carrying the genotype associated with higher dopamine receptor efficacy, i.e., A allele homozygotes of a single nucleotide polymorphism (rs907094 of the PPP1R1B gene. Moreover, this effect was more pronounced in children and older adults in comparison to adolescents and younger adults. Together, our findings indicate that polymorphic variations in a dopamine relevant gene are associated with individual differences in brain-evoked potentials of outcome updating and hint at the possibility that genotype effects on neurocognitive phenotypes may vary as a function of brain maturation and aging.

  18. NMDA and mGluR1 receptor subtypes as major players affecting depotentiation in the hippocampal CA1-region

    Amira Latif-Hernandez


    Full Text Available Neurons have the ability to modify their structure and function which ultimately serves for learning (Abraham and Bear, 1996. Dendritic events provide a major contribution to such modifications. For example, natural and artificial patterns of afferent activation have been shown to induce persistent forms of synaptic plasticity, such as long-term potentiation (LTP and long-term depression (LTD at distinct dendritic synapses. LTP and LTD are both assumed to occur during the physiological processes of learning and memory formation and to sustain the latter (Abraham, 2008. In recent years, there has been a burgeoning interest in the understanding of metaplasticity, which refers to the plasticity of synaptic plasticity (Abraham and Bear, 1996. In particular, depotentiation (DP is the mechanism by which synapses that have recently undergone LTP can reverse their synaptic strengthening in response to low frequency stimulation (LFS; Abraham, 2008. Typically, DP is thought to prevent the saturation of synaptic potentiation by resetting synapses into a more efficient state to store new information. The detailed mechanisms that underlie DP still remain unclear. Bortolotto et al. (1994 first identified metabotropic glutamate receptors (mGluRs as being involved in DP. Experimental evidence indicates that both subtypes of group I mGluRs (mGluR1 and mGluR5 have distinct functions in synaptic plasticity in the hippocampal CA1 region (Gladding et al., 2008. However, their role in DP was not addressed yet in detail and appear to be distinct from those involved in NMDAR-dependent DP (Zho et al., 2002. Therefore, we investigated the precise mechanisms responsible for NMDAR and mGluR-dependent DP by combining electrophysiological recordings in vitro and pharmacological approach. Transverse hippocampal slices (400 µm thick were prepared from the right hippocampus with a tissue chopper and placed into a submerged-type chamber, where they were continuously perfused

  19. RNA-Sequencing Reveals the Progression of Phage-Host Interactions between φR1-37 and Yersinia enterocolitica

    Katarzyna Leskinen; Blasdel, Bob G.; Rob Lavigne; Mikael Skurnik


    Despite the expanding interest in bacterial viruses (bacteriophages), insights into the intracellular development of bacteriophage and its impact on bacterial physiology are still scarce. Here we investigate during lytic infection the whole-genome transcription of the giant phage vB_YecM_φR1-37 (φR1-37) and its host, the gastroenteritis causing bacterium Yersinia enterocolitica. RNA sequencing reveals that the gene expression of φR1-37 does not follow a pattern typical observed in other lytic...

  20. High-density Association Mapping and Interaction Analysis of PLA2R1 and HLA Regions with Idiopathic Membranous Nephropathy in Japanese

    Thiri, Myo; Honda, Kenjiro; Kashiwase, Koichi; Mabuchi, Akihiko; Suzuki, Hodaka; WATANABE, KIMIO; Nakayama, Masaaki; Watanabe, Tsuyoshi; Doi, Kent; Tokunaga, Katsushi; Noiri, Eisei


    Although recent studies showed anti-PLA2R antibody plays a crucial role in idiopathic membranous nephropathy (IMN), detailed HLA mapping and interaction between the HLA genes and PLA2R1 have not been investigated in IMN. We genotyped across the PLA2R1 gene and the HLA region, using 183 IMN patients and 811 healthy controls. Five SNPs around the PLA2R1 gene were significantly associated with IMN. In addition to the two SNPs previously reported to be strongly associated with IMN, rs3749119 and ...

  1. Optimization of piperidin-4-yl-urea-containing melanin-concentrating hormone receptor 1 (MCH-R1) antagonists: Reducing hERG-associated liabilities.

    Berglund, Susanne; Egner, Bryan J; Gradén, Henrik; Gradén, Joakim; Morgan, David G A; Inghardt, Tord; Giordanetto, Fabrizio


    The discovery and optimization of piperidin-4-yl-urea derivatives as MCH-R1 antagonists is herein described. Previous work around the piperidin-4-yl-amides led to the discovery of potent MCH-R1 antagonists. However, high affinity towards the hERG potassium channel proved to be an issue. Different strategies to increase hERG selectivity were implemented and resulted in the identification of piperidin-4-yl-urea compounds as potent MCH-R1 antagonists with minimized hERG inhibition.

  2. An Exploratory Pilot Study of Genetic Marker for IgE-Mediated Allergic Diseases with Expressions of FcεR1α and Cε

    En-Chih Liao


    Full Text Available The high affinity immunoglobulin E (IgE receptor-FcεR1 is mainly expressed on the surface of effector cells. Cross-linking of IgE Abs bound to FcεR1 by multi-valent antigens can induce the activation of these cells and the secretion of inflammatory mediators. Since FcεR1 plays a central role in the induction and maintenance of allergic responses, this study aimed to investigate the association of FcεR1 with the allergic phenotype of Cε expression and cytokine and histamine release from peripheral leukocytes. Peripheral leukocytes from 67 allergic and 50 non-allergic subjects were used for genotyping analysis. Peripheral mononuclear cells (PBMCs were used for Cε expression and ELISpot analysis, while polymorphonuclear cells (PMNs were used for histamine release. The association between genotype polymorphism of the FcεR1α promoter region (rs2427827 and rs2251746 and allergic features of Cε expression and histamine were analyzed, and their effects on leukocytes function were compared with wild type. The genotype polymorphisms of FcεR1α promoter region with CT and TT in rs2427827 and TC in rs2251746 were significantly higher in allergic patients than in non-allergic controls. Patients with single nucleotide polymorphism (SNP of FcεR1α promoter region had high levels of total IgE, mite-specific Der p 2 (Group 2 allergen of Dermatophagoides pteronyssinus-specific IgE and IgE secretion B cells. The mRNA expression of FcεR1α was significantly increased after Der p2 stimulation in PBMCs with SNPs of the FcεR1α promoter region. Despite the increased Cε mRNA expression in PBMCs and histamine release from PMNs and the up-regulated mRNA expression of interleukin (IL-6 and IL-8 secretions after Der p2 stimulation, there was no statistically significant difference between SNPs of the FcεR1α promoter region and the wild type. SNPs of FcεR1α promoter region were associated with IgE expression, IgE producing B cells, and increased Der p2

  3. Birch pollen-related food allergy to legumes: identification and characterization of the Bet v 1 homologue in mungbean (Vigna radiata), Vig r 1.

    Mittag, D; Vieths, S; Vogel, L; Wagner-Loew, D; Starke, A; Hunziker, P; Becker, W-M; Ballmer-Weber, B K


    Recently allergic reactions to legumes mediated by Bet v 1-homologous food allergens were described for soy and peanut. In this study we assessed allergic reactions to another legume, to mungbean seedlings, and identified its Bet v 1-homologous allergen Vig r 1. Ten patients were selected who had a history of allergic reactions to mungbean seedlings and a respiratory allergy to birch pollen. The Bet v 1 homologue in mungbean seedlings, Vig r 1, was cloned by a PCR strategy, expressed in Escherichia coli, and purified by preparative SDS-PAGE. In all sera, specific IgE against birch pollen, Bet v 1, Bet v 2, Vig r 1, and the Bet v 1 homologues in soy (Gly m 4) and cherry (Pru av 1) was determined by CAP-FEIA. Cross-reactivity of specific IgE with Vig r 1, Bet v 1, Gly m 4, and Pru av 1 was assessed by immunoblot inhibition. Expression of Vig r 1 during development of mungbean seedlings and under wounding stress was analysed by immunoblotting. The Vig r 1 double band was analysed by matrix-assisted laser desorption/ionization time-of-flight and liquid chromatography/tandem mass spectrometry (LC/MS/MS). All patients were sensitized to birch pollen and Bet v 1, 20% to Bet v 2, and 90% to Gly m 4. Seventy percent of the patients showed IgE binding to a double band at 15 kDa in mungbean extract that was inhibited after pre-incubation of sera with rBet v 1. PCR cloning revealed that the mungbean homologue of Bet v 1 had a molecular weight of 16.2 kDa, a calculated pI of 4.6% and 42.8% amino acid sequence identity with Bet v 1. MS analysis confirmed similarity of the double band with the deduced Vig r 1 sequence, but also indicated the existence of other Vig r 1 isoforms. ImmunoCAP analysis detected IgE against Vig r 1 in 80% of the sera. IgE binding to Vig r 1 was inhibited with Gly m 4 in six of six and with rPru av 1 in four of six patients. Vig r 1 expression occurred during development of seedlings and was increased by wounding stress. Food allergy to mungbean

  4. The antibiotic thiostrepton inhibits a functional transition within protein L11 at the ribosomal GTPase centre

    Porse, B T; Leviev, I; Mankin, A S


    A newly identified class of highly thiostrepton-resistant mutants of the archaeon Halobacterium halobium carry a missense mutation at codon 18 within the gene encoding ribosomal protein L11. In the mutant proteins, a proline, conserved in archaea and bacteria, is converted to either serine or thr....... This putative inhibitory mechanism of thiostrepton is critically dependent on proline 18/22. Moreover, the absence of this proline from eukaryotic protein L11 sequences would account for the high thiostrepton resistance of eukaryotic ribosomes.......A newly identified class of highly thiostrepton-resistant mutants of the archaeon Halobacterium halobium carry a missense mutation at codon 18 within the gene encoding ribosomal protein L11. In the mutant proteins, a proline, conserved in archaea and bacteria, is converted to either serine...

  5. Development and implementation of a new pneumatic transfer system for materials irradiation at IEA-R1 reactor; Desenvolvimento e implementacao de um novo sistema pneumatico de transferencia para irradiacao de materiais no reator IEA-R1

    Fernando, Alberto de Jesus


    Pneumatic Transfer Systems (PTS) are classified as mechanical equipment largely operated all over the world for transport of a huge sort of objects, samples and materials located at nearly terminals or even at separated ones. System applicability is often recognized in many activities, such as medicine (hospital settings, clinical analysis labs), industry (steel, automobiles, mining, chemical, food, construction), trading (gas station, movies, supermarkets, banks, e-commerce) and federal agencies (post services, federal courts, public enterprises). In the nuclear settings, PTS shows also a vast array of applications, being a part of radioisotope production, as well as short-lived radiopharmaceuticals, including 67 Ga, 201 Tl, 18 F and 123 I-ultra pure. Besides, PTS are also used at radioactive waste management plants and research institutes that apply neutron activation analysis (NAA). This work was directed toward the design and operation of a new PTS for the IEA-R1 nuclear research reactor settled at Instituto de Pesquisas Energeticas e Nucleares (IPEN) for NAA application. With this aim, it was calculated the charge of reactor core grid plate and sample transport testing. Neutron flux at irradiating position was determined as 3,70 {+-} 0,26 10{sup 12} n cm{sup -2} s{sup -1}. (author)

  6. Optimization of the irradiation beam in the BNCT research facility at IEA-R1 reactor; Otimizacao do feixe de irradiacao na instalacao para estudos em BNCT junto ao reator IEA-R1

    Castro, Vinicius Alexandre de


    Boron Neutron Capture Therapy (BNCT) is a radiotherapeutic technique for the treatment of some types of cancer whose useful energy comes from a nuclear reaction that occurs when thermal neutron impinges upon a Boron-10 atom. In Brazil there is a research facility built along the beam hole number 3 of the IEA-R1 research reactor at IPEN, which was designed to perform BNCT research experiments. For a good performance of the technique, the irradiation beam should be mostly composed of thermal neutrons with a minimum as possible gamma and above thermal neutron components. This work aims to monitor and evaluate the irradiation beam on the sample irradiation position through the use of activation detectors (activation foils) and also to propose, through simulation using the radiation transport code, MCNP, new sets of moderators and filters which shall deliver better irradiation fields at the irradiation sample position In this work, a simulation methodology, based on a MCNP card, known as wwg (weight window generation) was studied, and the neutron energy spectrum has been experimentally discriminated at 5 energy ranges by using a new set o activation foils. It also has been concluded that the BNCT research facility has the required thermal neutron flux to perform studies in the area and it has a great potential for improvement for tailoring the irradiation field. (author)

  7. Expression of the AMPA Receptor Subunits GluR1 and GluR2 is Associated with Granule Cell Maturation in the Dentate Gyrus

    Hagihara, Hideo; Ohira, Koji; Toyama, Keiko; Miyakawa, Tsuyoshi


    The dentate gyrus produces new granule neurons throughout adulthood in mammals from rodents to humans. During granule cell maturation, defined markers are expressed in a highly regulated sequential process, which is necessary for directed neuronal differentiation. In the present study, we show that α-amino-3-hydroxy-5-methy-4-isoxazole propionate (AMPA) receptor subunits GluR1 and GluR2 are expressed in differentiated granule cells, but not in stem cells, in neonatal, and adult dentate gyrus. Using markers for neural progenitors, immature and mature granule cells, we found that GluR1 and GluR2 were expressed mainly in mature cells and in some immature cells. A time-course analysis of 5-bromo-2′-deoxyuridine staining revealed that granule cells express GluR1 around 3 weeks after being generated. In mice heterozygous for the alpha-isoform of calcium/calmodulin-dependent protein kinase II, a putative animal model of schizophrenia and bipolar disorder in which dentate gyrus granule cells fail to mature normally, GluR1 and GluR2 immunoreactivities were substantially downregulated in the dentate gyrus granule cells. In the granule cells of mutant mice, the expression of both presynaptic and postsynaptic markers was decreased, suggesting that GluR1 and GluR2 are also associated with synaptic maturation. Moreover, GluR1 and GluR2 were also expressed in mature granule cells of the neonatal dentate gyrus. Taken together, these findings indicate that GluR1 and GluR2 expression closely correlates with the neuronal maturation state, and that GluR1 and GluR2 are useful markers for mature granule cells in the dentate gyrus. PMID:21927594

  8. Molecular identification of ghrelin receptor (GHS-R1a) and its functional role in the gastrointestinal tract of the guinea-pig.

    Kitazawa, Takio; Nakamura, Tatsuro; Saeki, Atsuki; Teraoka, Hiroki; Hiraga, Takeo; Kaiya, Hiroyuki


    Ghrelin stimulates gastric motility in vivo in the guinea-pig through activation of growth hormone secretagogue receptor (GHS-R). In this study, we identified GHS-R1a in the guinea-pig, and examined its distribution and cellular function and compared them with those in the rat. Effects of ghrelin in different regions of gastrointestinal tract were also examined. GHS-R1a was identified in guinea-pig brain cDNA. Amino acid identities of guinea-pig GHS-R1a were 93% to horses and 85% to dogs. Expression levels of GHS-R1a mRNA were high in the pituitary and hypothalamus, moderate in the thalamus, cerebral cortex, pons, medulla oblongata and olfactory bulb, and low in the cerebellum and peripheral tissues including gastrointestinal tract. Comparison of GHS-R1a expression patterns showed that those in the brain were similar but the expression level in the gastrointestinal tract was higher in rats than in guinea-pigs. Guinea-pig GHS-R1a expressed in HEK 293 cells responded to rat ghrelin and GHS-R agonists. Rat ghrelin was ineffective in inducing mechanical changes in the stomach and colon but caused a slight contraction in the small intestine. 1,1-Dimethyl-4-phenylpiperazinium and electrical field stimulation (EFS) caused cholinergic contraction in the intestine, and these contractions were not affected by ghrelin. Ghrelin did not change spontaneous and EFS-evoked [(3)H]-efflux from [(3)H]-choline-loaded ileal strips. In summary, guinea-pig GHS-R1a was identified and its functions in isolated gastrointestinal strips were characterized. The distribution of GHS-R1a in peripheral tissues was different from that in rats, suggesting that the functional role of ghrelin in the guinea-pig is different from that in other animal species.

  9. PLA2R-associated membranous glomerulopathy is modulated by common variants in PLA2R1 and HLA-DQA1 genes.

    Saeed, M; Beggs, M L; Walker, P D; Larsen, C P


    Membranous glomerulopathy (MG) is most commonly caused by autoantibodies directed against the podocyte phospholipase A2 receptor (PLA2R1) and common variants in this gene are associated with MG. Here for the first time, we carried out a large case-control association study (n=1512) of PLA2R-positive and -negative MG to determine the extent of association in these pathologic subtypes. We performed four separate sets of analyses to determine significance of the single-nucleotide polymorphisms (SNPs) and their haplotypes followed by joint analysis and trans-ethnic mapping to increase power. The PLA2R1 SNP rs35771982 was most strongly associated with PLA2R-positive MG (P=1.4 × 10(-14), odds ratio (ORGG)=1.98). The associations of other SNPs in PLA2R1 could be explained because of linkage disequilibrium with the G-allele. Haplotypes in PLA2R1 did not exceed the significance of rs35771982 even after 10 000 permutations. PLA2R1 variants were only associated with PLA2R-positive MG and predominantly in Caucasians. PLA2R1 variants did not associate with MG in African Americans (AA). There was strong epistasis between HLA-DQA1 SNP rs2187668 and the PLA2R1 variant rs35771982. Thus, common variants in the PLA2R1, particularly rs35771982, modulate PLA2R-positive MG with HLA-DQA1 in Caucasians. PLA2R-negative MG especially in AA, may provide a novel opportunity to discover new genes underlying MG.

  10. Pseudogenization of the umami taste receptor gene Tas1r1 in the giant panda coincided with its dietary switch to bamboo.

    Zhao, Huabin; Yang, Jian-Rong; Xu, Huailiang; Zhang, Jianzhi


    Although it belongs to the order Carnivora, the giant panda is a vegetarian with 99% of its diet being bamboo. The draft genome sequence of the giant panda shows that its umami taste receptor gene Tas1r1 is a pseudogene, prompting the proposal that the loss of the umami perception explains why the giant panda is herbivorous. To test this hypothesis, we sequenced all six exons of Tas1r1 in another individual of the giant panda and five other carnivores. We found that the open reading frame (ORF) of Tas1r1 is intact in all these carnivores except the giant panda. The rate ratio (ω) of nonsynonymous to synonymous substitutions in Tas1r1 is significantly higher for the giant panda lineage than for other carnivore lineages. Based on the ω change and the observed number of ORF-disrupting substitutions, we estimated that the functional constraint on the giant panda Tas1r1 was relaxed ∼ 4.2 Ma, with its 95% confidence interval between 1.3 and 10 Ma. Our estimate matches the approximate date of the giant panda's dietary switch inferred from fossil records. It is probable that the giant panda's decreased reliance on meat resulted in the dispensability of the umami taste, leading to Tas1r1 pseudogenization, which in turn reinforced its herbivorous life style because of the diminished attraction of returning to meat eating in the absence of Tas1r1. Nonetheless, additional factors are likely involved because herbivores such as cow and horse still retain an intact Tas1r1.

  11. Loss of the oncogenic phosphatase PRL-3 promotes a TNF-R1 feedback loop that mediates triple-negative breast cancer growth.

    Gari, H H; DeGala, G D; Lucia, M S; Lambert, J R


    Stimulating tumor cell senescence and apoptosis are proven methods for therapeutically combating cancer. However, senescence and apoptosis are conventionally viewed as parallel, not sequential, processes. We have discovered that the metastasis-promoting phosphatase, PRL-3, is transcriptionally regulated by the NF-ĸB pathway in triple-negative breast cancer (TNBC) cells, and that PRL-3 knockdown elicits an autocrine tumor necrosis factor receptor 1 (TNF-R1) feedback loop that results in TNBC cell senescence followed by apoptosis. Knockdown of PRL-3 leads to rapid G1 cell cycle arrest and induction of a strong TNFα cytokine response that promotes a period of cellular senescence through TNF-R1-mediated activation of NF-ĸB. Senescent PRL-3 knockdown cells subsequently underwent apoptosis as a result of increased TNF-R1 signaling through the TNFα-associated extrinsic death pathway, shunting signaling away from the NF-ĸB cascade. These data suggest that TNF-R1 signaling dynamically re-programs after PRL-3 knockdown, from sustaining cell senescence through NF-ĸB to promoting apoptosis through TNF-R1 internalization and caspase-8 activation. The molecular mechanisms that determine the survival-death balance of TNF-R1 signaling are poorly understood, despite the fact that TNF-R1 has been extensively studied. Our results describe PRL-3 knockdown as a novel survival-death balance modifier of the TNF-R1 pathway, and show that senescent TNBC tumor cells can be sensitized to undergo apoptosis in a sequential manner.

  12. Does DcR1 (TNF-related apoptosis-inducing-ligand Receptor 3) have any role in human AMD pathogenesis?

    Anand, Akshay; Sharma, Neel K; Singh, Ramandeep; Gupta, Amod; Prabhakar, Sudesh; Jindal, Neeru; Bhatt, Arvind K; Sharma, Suresh K; Gupta, Pawan K


    It has been postulated that there is a link between age related degenerative diseases and cancer. The TNF-related apoptosis-inducing ligand (TRAIL) has been shown to selectively kill tumor cells by binding to pro-apoptotic and anti-apoptotic receptors. Our aim was to study the levels of anti-apoptotic receptor (DcR1) in age related macular degeneration (AMD) and controls. AMD patients (115) were classified into two groups: Dry and Wet AMD. Wet AMDs were further classified into occult, predominant classic and minimal classic. 61 healthy individuals were recruited as normal controls. After normalization with total protein, DcR1 levels were analyzed by ELISA. Mann Whitney U-statistic was used for analysis of DcR1 ELISA results. We have observed DcR1 levels in serum sample which were significantly lower in AMD patients as compared to controls (p = 0.001). On the other hand, we did not find difference in DcR1 levels between wet and dry AMD. The present study defines the plausible role of DcR1 in AMD pathology signifying a new therapeutic target for AMD.

  13. Ligand specificity and affinity of BT-R1, the Bacillus thuringiensis Cry1A toxin receptor from Manduca sexta, expressed in mammalian and insect cell cultures.

    Keeton, T P; Bulla, L A


    The Manduca sexta receptor for the Bacillus thuringiensis Cry1Aa, Cry1Ab, and Cry1Ac toxins, BT-R1, has been expressed in heterologous cell culture, and its ligand binding characteristics have been determined. When transfected with the BT-R1 cDNA, insect and mammalian cell cultures produce a binding protein of approximately 195 kDa, in contrast to natural BT-R1 from M. sexia, which has an apparent molecular weight of 210 kDa. Transfection of cultured Spodoptera frugiperda cells with the BT-R1 cDNA imparts Cry1A-specific high-affinity binding activity typical of membranes prepared from larval M. sexta midguts. Competition assays with BT-R1 prepared from larval M. sexta midguts and transiently expressed in cell culture reveal virtually identical affinities for the Cry1Aa, Cry1Ab, and Cry1Ac toxins, clearly demonstrating the absolute specificity of the receptor for toxins of the lepidopteran-specific Cry1A family. BT-R1 therefore remains the only M. sexta Cry1A binding protein to be purified, cloned, and functionally expressed in heterologous cell culture, and for the first time, we are able to correlate the Cry1Aa, Cry1Ab, and Cry1Ac toxin sensitivities of M. sexta to the identity and ligand binding characteristics of a single midgut receptor molecule. PMID:9292994

  14. Independent activity of the homologous small regulatory RNAs AbcR1 and AbcR2 in the legume symbiont Sinorhizobium meliloti.

    Omar Torres-Quesada

    Full Text Available The legume symbiont Sinorhizobium meliloti expresses a plethora of small noncoding RNAs (sRNAs whose function is mostly unknown. Here, we have functionally characterized two tandemly encoded S. meliloti Rm1021 sRNAs that are similar in sequence and structure. Homologous sRNAs (designated AbcR1 and AbcR2 have been shown to regulate several ABC transporters in the related α-proteobacteria Agrobacterium tumefaciens and Brucella abortus. In Rm1021, AbcR1 and AbcR2 exhibit divergent unlinked regulation and are stabilized by the RNA chaperone Hfq. AbcR1 is transcribed in actively dividing bacteria, either in culture, rhizosphere or within the invasion zone of mature alfalfa nodules. Conversely, AbcR2 expression is induced upon entry into stationary phase and under abiotic stress. Only deletion of AbcR1 resulted into a discrete growth delay in rich medium, but both are dispensable for symbiosis. Periplasmic proteome profiling revealed down-regulation of the branched-chain amino acid binding protein LivK by AbcR1, but not by AbcR2. A double-plasmid reporter assay confirmed the predicted specific targeting of the 5'-untranslated region of the livK mRNA by AbcR1 in vivo. Our findings provide evidences of independent regulatory functions of these sRNAs, probably to fine-tune nutrient uptake in free-living and undifferentiated symbiotic rhizobia.

  15. Bub3-BubR1-dependent sequestration of Cdc20Fizzy at DNA breaks facilitates the correct segregation of broken chromosomes.

    Derive, Nicolas; Landmann, Cedric; Montembault, Emilie; Claverie, Marie-Charlotte; Pierre-Elies, Priscillia; Goutte-Gattat, Damien; Founounou, Nabila; McCusker, Derek; Royou, Anne


    The presence of DNA double-strand breaks during mitosis is particularly challenging for the cell, as it produces broken chromosomes lacking a centromere. This situation can cause genomic instability resulting from improper segregation of the broken fragments into daughter cells. We recently uncovered a process by which broken chromosomes are faithfully transmitted via the BubR1-dependent tethering of the two broken chromosome ends. However, the mechanisms underlying BubR1 recruitment and function on broken chromosomes were largely unknown. We show that BubR1 requires interaction with Bub3 to localize on the broken chromosome fragments and to mediate their proper segregation. We also find that Cdc20, a cofactor of the E3 ubiquitin ligase anaphase-promoting complex/cyclosome (APC/C), accumulates on DNA breaks in a BubR1 KEN box-dependent manner. A biosensor for APC/C activity demonstrates a BubR1-dependent local inhibition of APC/C around the segregating broken chromosome. We therefore propose that the Bub3-BubR1 complex on broken DNA inhibits the APC/C locally via the sequestration of Cdc20, thus promoting proper transmission of broken chromosomes.

  16. MadR1, a Mycobacterium tuberculosis cell cycle stress response protein that is a member of a widely conserved protein class of prokaryotic, eukaryotic and archeal origin.

    Crew, Rebecca; Ramirez, Melissa V; England, Kathleen; Slayden, Richard A


    Stress-induced molecular programs designed to stall division progression are nearly ubiquitous in bacteria, with one well-known example being the participation of the SulA septum inhibiting protein in the SOS DNA damage repair response. Mycobacteria similarly demonstrate stress-altered growth kinetics, however no such regulators have been found in these organisms. We therefore set out to identify SulA-like regulatory proteins in Mycobacterium tuberculosis. A bioinformatics modeling-based approach led to the identification of rv2216 as encoding for a protein with weak similarity to SulA, further analysis distinguished this protein as belonging to a group of uncharacterized growth promoting proteins. We have named the mycobacterial protein encoded by rv2216 morphology altering division regulator protein 1, MadR1. Overexpression of madR1 modulated cell length while maintaining growth kinetics similar to wild-type, and increased the proportion of bent or V-form cells in the population. The presence of MadR1-GFP at regions of cellular elongation (poles) and morphological differentiation (V-form) suggests MadR1 involvement in phenotypic heterogeneity and longitudinal cellular growth. Global transcriptional analysis indicated that MadR1 functionality is linked to lipid editing programs required for growth and persistence. This is the first report to differentiate the larger class of these conserved proteins from SulA proteins and characterizes MadR1 effects on the mycobacterial cell.

  17. Independent Activity of the Homologous Small Regulatory RNAs AbcR1 and AbcR2 in the Legume Symbiont Sinorhizobium meliloti

    Torres-Quesada, Omar; Millán, Vicenta; Nisa-Martínez, Rafael; Bardou, Florian; Crespi, Martín; Toro, Nicolás; Jiménez-Zurdo, José I.


    The legume symbiont Sinorhizobium meliloti expresses a plethora of small noncoding RNAs (sRNAs) whose function is mostly unknown. Here, we have functionally characterized two tandemly encoded S. meliloti Rm1021 sRNAs that are similar in sequence and structure. Homologous sRNAs (designated AbcR1 and AbcR2) have been shown to regulate several ABC transporters in the related α-proteobacteria Agrobacterium tumefaciens and Brucella abortus. In Rm1021, AbcR1 and AbcR2 exhibit divergent unlinked regulation and are stabilized by the RNA chaperone Hfq. AbcR1 is transcribed in actively dividing bacteria, either in culture, rhizosphere or within the invasion zone of mature alfalfa nodules. Conversely, AbcR2 expression is induced upon entry into stationary phase and under abiotic stress. Only deletion of AbcR1 resulted into a discrete growth delay in rich medium, but both are dispensable for symbiosis. Periplasmic proteome profiling revealed down-regulation of the branched-chain amino acid binding protein LivK by AbcR1, but not by AbcR2. A double-plasmid reporter assay confirmed the predicted specific targeting of the 5′-untranslated region of the livK mRNA by AbcR1 in vivo. Our findings provide evidences of independent regulatory functions of these sRNAs, probably to fine-tune nutrient uptake in free-living and undifferentiated symbiotic rhizobia. PMID:23869210

  18. UV-dependent production of 25-hydroxyvitamin D{sub 2} in the recombinant yeast cells expressing human CYP2R1

    Yasuda, Kaori; Endo, Mariko; Ikushiro, Shinichi; Kamakura, Masaki [Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398 (Japan); Ohta, Miho [Department of Food and Nutrition Management Studies, Faculty of Human Development, Soai University, 4-4-1 Nanko-naka, Suminoe-ku, Osaka 559-0033 (Japan); Sakaki, Toshiyuki, E-mail: [Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398 (Japan)


    Highlights: •We produce 25-hydroxyvitamin D in the recombinant yeast expressing human CYP2R1. •Vitamin D2 is produced in yeast from endogenous ergosterol with UV irradiation. •We produce 25-hydroxyvitamin D2 in the recombinant yeast without added substrate. -- Abstract: CYP2R1 is known to be a physiologically important vitamin D 25-hydroxylase. We have successfully expressed human CYP2R1 in Saccharomyces cerevisiae to reveal its enzymatic properties. In this study, we examined production of 25-hydroxylated vitamin D using whole recombinant yeast cells that expressed CYP2R1. When vitamin D{sub 3} or vitamin D{sub 2} was added to the cell suspension of CYP2R1-expressing yeast cells in a buffer containing glucose and β-cyclodextrin, the vitamins were converted into their 25-hydroxylated products. Next, we irradiated the cell suspension with UVB and incubated at 37 °C. Surprisingly, the 25-hydroxy vitamin D{sub 2} was produced without additional vitamin D{sub 2}. Endogenous ergosterol was likely converted into vitamin D{sub 2} by UV irradiation and thermal isomerization, and then the resulting vitamin D{sub 2} was converted to 25-hydroxyvitamin D{sub 2} by CYP2R1. This novel method for producing 25-hydroxyvitamin D{sub 2} without a substrate could be useful for practical purposes.

  19. Prognosis of R1-resection at the bronchial stump in patients with non-small cell lung cancer

    Lyu Jima; Hao Xuezhi; Hui Zhouguang; Liang Jun; Zhou Zongmei; Feng Qinfu; Xiao Zefen


    Background The prognosis of R1-resection at the bronchial stump in patients with non-small cell lung cancer (NSCLC) remains unclear.This study intends to identify the prognostic factors and to optimize treatments for these patients under update conditions.Methods The data of 124 NSCLC patients who underwent R1-resection at the bronchial stump was reviewed.There were 41 patients in the surgery group (S),21 in the postoperative radiotherapy (PORT) group (S+R),30 in the postoperative chemotherapy (POCT) group (S+C),and 32 in the PORT plus POCT group (S+R+C).The constitute proportion in different groups was tested using the X2 method,univariate analysis was performed using the Kaplan-Meier and log-rank method,and multivariate analysis was done using the Cox hazard regression with entry factors including age,sex,pathological type and stage,classification of the residual disease,and treatment procedure.The process was performed stepwise backward with a maximum iteration of 20 and an entry possibility of 0.05 as well as an excluded possibility of 0.10 at each step.Results In univariate analysis,survival was more favorable for patients with squamous cell carcinoma,early pathological T or N stage,and chemotherapy or radiotherapy.There was no significant difference in the survival for patients with different types of the residual disease,except for the difference between patients with carcinoma in situ and lymphangiosis carcinomatosa (P=0.030).The survival for patients receiving chemoradiotherapy was superior to that for those undergoing surgery alone (P=0.016).In multivariate analysis,the pathological type (HR 2.51,95% CI 1.59 to 3.96,P=0.000),pathological T (HR 1.29,95% CI 1.04 to 1.60,P=-0.021) or N stage (HR 2.04,95% CI 1.40 to 2.98,P=0.000),and chemotherapy (HR 0.24,95% CI 0.13 to 0.43,P=0.000) were independent prognostic factors.Conclusion Patients with squamous cell carcinoma,early pathological T or N stage,or receiving chemotherapy had a more favorable

  20. C/2013 R1 (Lovejoy) at IR wavelengths and the variability of CO abundances among Oort Cloud comets

    Paganini, L.; Mumma, M. J.; Villanueva, G. L.; Bonev, B. P.; DiSanti, M. A. [Goddard Center for Astrobiology, NASA GSFC, MS 690, Greenbelt, MD 20771 (United States); Keane, J. V.; Meech, K. J. [Institute for Astronomy, University of Hawaii, Honolulu, HI 96822 (United States); Blake, G. A. [Division of Geological and Planetary Sciences, California Institute of Technology, Pasadena, CA 91125 (United States); Gibb, E. L., E-mail: [Department of Physics and Astronomy, University of Missouri-St. Louis, St. Louis, MO 63121 (United States)


    We report production rates, rotational temperatures, and related parameters for gases in C/2013 R1 (Lovejoy) using the Near InfraRed SPECtrometer at the Keck Observatory, on six UT dates spanning heliocentric distances (R{sub h} ) that decreased from 1.35 AU to 1.16 AU (pre-perihelion). We quantified nine gaseous species (H{sub 2}O, OH*, CO, CH{sub 4}, HCN, C{sub 2}H{sub 6}, CH{sub 3}OH, NH{sub 3}, and NH{sub 2}) and obtained upper limits for two others (C{sub 2}H{sub 2} and H{sub 2}CO). Compared with organics-normal comets, our results reveal highly enriched CO, (at most) slightly enriched CH{sub 3}OH, C{sub 2}H{sub 6}, and HCN, and CH{sub 4} consistent with {sup n}ormal{sup ,} yet depleted, NH{sub 3}, C{sub 2}H{sub 2}, and H{sub 2}CO. Rotational temperatures increased from ∼50 K to ∼70 K with decreasing R{sub h} , following a power law in R{sub h} of –2.0 ± 0.2, while the water production rate increased from 1.0 to 3.9 × 10{sup 28} molecules s{sup –1}, following a power law in R{sub h} of –4.7 ± 0.9. The ortho-para ratio for H{sub 2}O was 3.01 ± 0.49, corresponding to spin temperatures (T {sub spin}) ≥ 29 K (at the 1σ level). The observed spatial profiles for these emissions showed complex structures, possibly tied to nucleus rotation, although the cadence of our observations limits any definitive conclusions. The retrieved CO abundance in Lovejoy is more than twice the median value for comets in our IR survey, suggesting this comet is enriched in CO. We discuss the enriched value for CO in comet C/2013 R1 in terms of the variability of CO among Oort Cloud comets.

  1. An Extended Istomin-Palm Model for Estimating the Enthalpies of Formation of the Two-Direction Extending Compounds R1-Y-R2%用扩展的Istomin-Palm模型估算双向延伸化合物R1-Y-R2生成焓

    曹晨忠; 武亚新


    Istomin and Palm proposed a model, ΔfH0(RX)=h[R]+h[X]+φ[R]φ[X], (h[R] and h[X] are the contributions of alkyl R and substituent X to theΔfH0(RX), respectively.φ[R]φ[X] represents the interaction of alkyl R and substituent X), to express the enthalpies of formation of monoderivatives of hydrocarbonsΔfH0(RX). However, in two-direction extending compounds R1-Y-R2, the Y substituent is attached to two alkyl groups (R1 and R2), and the intramolecular interactions are more complicated than that in monosubstituted alkanes. Thus, the Istomin-Palm model must be modified. In this work, the interactions among Y, R1, and R2 contributing to the enthalpy of formation,ΔfH0(R1-Y-R2), are divided into three parts: the interaction between R1Y and R2 (φ[R2]φ[R1Y]), the interaction between YR2 and R1 (φ[R1]φ[YR2]), and the interaction between R1 and R2 (ψ[R1]ψ[R2]). These three interactions replace theφ[R]φ[X] term, and a new extended Istomin-Palm model,ΔfH0(R1-Y-R2)=h[R1]+h[R2]+h[Y]+φ[R1]φ[YR2]+φ[R2]φ[R1Y]+ψ[R1]ψ[R2], is proposed. In this model, h[Y] is the contribution of substituent Y to ΔfH0(R1-Y-R2). The h[R1] and h[R2] terms are the contributions of alkyls R1 and R2 toΔfH0(R1-Y-R2). The last three terms are the total contribution of interactions among Y, R1, and R2. Furthermore, the interaction potential index IPI(X) reported in our recent work (Wu, Y. X.; Cao, C. Z.; Yuan, H. Chin. J. Chem. Phys. 2012, 25 (2), 153.) was employed to express the intrinsic interaction of Y with alkyl groups (φ[Y]), and two general expressions were established to estimate ΔfH0, in which one is for thioethers, secondary amines, ethers, and ketones, and the other is for esters. These two estimating equations give results, which are as accurate as G3 and G3MP2 models in calculating ΔfH0 for R1-Y-R2 compounds. Moreover, our method avoids time consuming calculations.%  Istomin和Palm曾提出用模型ΔfH0(RX)=h[R]+h[X]+φ[R]φ[X](式中h[R]和h[X]分别为烷基R和取

  2. Myocilin is involved in NgR1/Lingo-1-mediated oligodendrocyte differentiation and myelination of the optic nerve.

    Kwon, Heung Sun; Nakaya, Naoki; Abu-Asab, Mones; Kim, Hong Sug; Tomarev, Stanislav I


    Myocilin is a secreted glycoprotein that belongs to a family of olfactomedin domain-containing proteins. Although myocilin is detected in several ocular and nonocular tissues, the only reported human pathology related to mutations in the MYOCILIN gene is primary open-angle glaucoma. Functions of myocilin are poorly understood. Here we demonstrate that myocilin is a mediator of oligodendrocyte differentiation and is involved in the myelination of the optic nerve in mice. Myocilin is expressed and secreted by optic nerve astrocytes. Differentiation of optic nerve oligodendrocytes is delayed in Myocilin-null mice. Optic nerves of Myocilin-null mice contain reduced levels of several myelin-associated proteins including myelin basic protein, myelin proteolipid protein, and 2'3'-cyclic nucleotide 3'-phosphodiesterase compared with those of wild-type littermates. This leads to reduced myelin sheath thickness of optic nerve axons in Myocilin-null mice compared with wild-type littermates, and this difference is more pronounced at early postnatal stages compared with adult mice. Myocilin also affects differentiation of oligodendrocyte precursors in vitro. Its addition to primary cultures of differentiating oligodendrocyte precursors increases levels of tested markers of oligodendrocyte differentiation and stimulates elongation of oligodendrocyte processes. Myocilin stimulation of oligodendrocyte differentiation occurs through the NgR1/Lingo-1 receptor complex. Myocilin physically interacts with Lingo-1 and may be considered as a Lingo-1 ligand. Myocilin-induced elongation of oligodendrocyte processes may be mediated by activation of FYN and suppression of RhoA GTPase.

  3. Study of human factors and its basic aspects, focusing the operators of IEA-R1 research reactor

    Martins, Maria da Penha Sanches; Andrade, Delvonei Alves de [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)]. E-mails:;


    The objective of this work is the study of human factors and situational variables, which, when modified, can interfere in the work actions of the operators of nuclear installations. This work is focused on the operators of the IEA-R1 research reactor, which is located in the Instituto de Pesquisas Energeticas e Nucleares - IPEN - CNEN/SP. The accidents in Nuclear Plants have shown that the most serious have occurred due to human failure. This work also considers the item 5.5.3 of CNEN-NN-3.01 standard - 'Actions must be taken to reduce, as much as possible, the human failures that may lead to accidents or even other events which may originate inadvertent or unintentional expositions in any individual'. The model named - Behavioral Analysis - is adopted. Relevant factors and aspects of the operators' routine are also considered. It is worth to remind that the performance depends on a series of variables, not only on the individual, but also the situational ones, which include physical, work, environment, organizational and social variables. Subjective factors are also considered, such as: attitude, ability, motivation etc., aiming at a global perspective of the situation, which counts on a set of principles for the behavior analysis and comprehension. After defining the applicability scenario, mechanisms and corrective actions to contribute with the reduction of failures will be proposed. (author)

  4. Study of human factors and its basic aspects, focusing the operators of IEA-R1 research reactor

    Martins, Maria da Penha Sanches; Andrade, Delvonei Alves de [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)]. E-mails:;


    Human factors and situational variables, which ca, when modified, interfere in the actions of operators of nuclear installations is studied. This work is focused in the operators of the IEA-R1 research reactor, which is located in the Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Brazil. The accidents in Nuclear Plants have shown that the most serious have occurred due to human failure. This work also considers the item 5.5.3 of CNEN-NN-3.01 standard - 'Actions must be taken to reduce, as much as possible, the human failures that can lead to accidents or even other events which can originate inadvertent or unintentional expositions in any individual'. The model named 'Behavioral Analysis' is adopted. Relevant factors and aspects of the operators' routine are also considered. It is worth to remind that the performance depends on a series of variables, not only on the individual, but also situational, including in these categories; physical variables, work environment, organizational and the social ones. The subjective factors are also considered, such as: attitude, ability, motivation etc., aiming at a global perspective of the situation, which counts on a set of principles for the behaviour analysis and comprehension. After defining the applicability scenario, mechanisms and corrective actions to contribute with the reduction of failures will be proposed. (author)

  5. Bioinformatics Analysis of the FREM1 Gene—Evolutionary Development of the IL-1R1 Co-Receptor, TILRR

    Eva E. Qwarnstrom


    Full Text Available The TLRs and IL-1 receptors have evolved to coordinate the innate immune response following pathogen invasion. Receptors and signalling intermediates of these systems are generally characterised by a high level of evolutionary conservation. The recently described IL-1R1 co-receptor TILRR is a transcriptional variant of the FREM1 gene. Here we investigate whether innate co-receptor differences between teleosts and mammals extend to the expression of the TILRR isoform of FREM1. Bioinformatic and phylogenetic approaches were used to analyse the genome sequences of FREM1 from eukaryotic organisms including 37 tetrapods and five teleost fish. The TILRR consensus peptide sequence was present in the FREM1 gene of the tetrapods, but not in fish orthologs of FREM1, and neither FREM1 nor TILRR were present in invertebrates. The TILRR gene appears to have arisen via incorporation of adjacent non-coding DNA with a contiguous exonic sequence after the teleost divergence. Comparing co-receptors in other systems, points to their origin during the same stages of evolution. Our results show that modern teleost fish do not possess the IL-1RI co-receptor TILRR, but that this is maintained in tetrapods as early as amphibians. Further, they are consistent with data showing that co-receptors are recent additions to these regulatory systems and suggest this may underlie differences in innate immune responses between mammals and fish.

  6. TRAPPIST photometry and imaging monitoring of comet C/2013 R1(Lovejoy): Implications for the origin of daughter species

    Opitom, C; Manfroid, J; Hutsemékers, D; Gillon, M; Magain, P


    We report the results of the narrow band photometry and imaging monitoring of comet C/2013 R1 (Lovejoy) with the robotic telescope TRAPPIST (La Silla observatory). We gathered around 400 images over 8 months pre- and post-perihelion between September 12, 2013 and July 6, 2014. We followed the evolution of the OH, NH, CN, C3 , and C2 production rates computed with the Haser model as well as the evolution of the dust production. All five gas species display an asymmetry about perihelion, the rate of brightening being steeper than the rate of fading. The study of the coma morphology reveals gas and dust jets which indicate one or several active zone(s) on the nucleus. The dust, C2 , and C3 morphologies present some similarities while the CN morphology is different. OH and NH are enhanced in the tail direction. The study of the evolution of the comet activity shows that the OH, NH, and C2 production rates evolution with the heliocentric distance is correlated to the dust evolution. The CN and, to a lesser extent,...

  7. 23(S),25(R)-1,25-dihydroxyvitamin D3-26,23-lactone stimulates murine bone formation in vivo

    Shima, M.; Tanaka, H.; Norman, A.W.; Yamaoka, K.; Yoshikawa, H.; Takaoka, K.; Ishizuka, S.; Seino, Y. (Osaka Univ. School of Medicine (Japan))


    23(S),25(R)-1,25-Dihydroxyvitamin D3-26,23-lactone (1,25-lactone) has been shown to have unique actions different from those of 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). In contrast to 1,25-(OH)2D3, 1,25-lactone causes a significant reduction in the serum Ca2+ level, stimulates collagen production in an osteoblastic cell line, and inhibits bone resorption induced by 1,25-(OH)2D3. A possible effect of 1,25-lactone on bone formation was examined in experiments on ectopic bone formation using a bone-inducing factor derived from Dunn osteosarcomas. 1,25-Lactone, a metabolite of 1,25-(OH)2D3, increased (3H)proline uptake at the stage of chondrogenesis and {sup 85}Sr uptake during bone formation. Significantly enlarged bone was also induced by this compound 3 weeks after implantation. These results suggest that the 1,25-lactone may be able to stimulate bone formation under in vivo conditions.

  8. Association of exposure to manganese and iron with relaxation rates R1 and R2*- magnetic resonance imaging results from the WELDOX II study.

    Pesch, Beate; Dydak, Ulrike; Lotz, Anne; Casjens, Swaantje; Quetscher, Clara; Lehnert, Martin; Abramowski, Jessica; Stewig, Christoph; Yeh, Chien-Lin; Weiss, Tobias; van Thriel, Christoph; Herrmann, Lennard; Muhlack, Siegfried; Woitalla, Dirk; Glaubitz, Benjamin; Schmidt-Wilcke, Tobias; Brüning, Thomas


    Magnetic resonance imaging is a non-invasive method that allows the indirect quantification of manganese (Mn) and iron (Fe) accumulation in the brain due to their paramagnetic features. The WELDOX II study aimed to explore the influence of airborne and systemic exposure to Mn and Fe on the brain deposition using the relaxation rates R1 and R2* as biomarkers of metal accumulation in regions of interest in 161 men, including active and former welders. We obtained data on the relaxation rates R1 and R2* in regions that included structures within the globus pallidus (GP), substantia nigra (SN), and white matter of the frontal lobe (FL) of both hemispheres, as well as Mn in whole blood (MnB), and serum ferritin (SF). The study subjects, all male, included 48 active and 20 former welders, 41 patients with Parkinson's disease (PD), 13 patients with hemochromatosis (HC), and 39 controls. Respirable Mn and Fe were measured during a working shift for welders. Mixed regression models were applied to estimate the effects of MnB and SF on R1 and R2*. Furthermore, we estimated the influence of airborne Mn and Fe on the relaxation rates in active welders. MnB and SF were significant predictors of R1 but not of R2* in the GP, and were marginally associated with R1 in the SN (SF) and FL (MnB). Being a welder or suffering from PD or HC elicited no additional group effect on R1 or R2* beyond the effects of MnB and SF. In active welders, shift concentrations of respirable Mn>100μg/m(3) were associated with stronger R1 signals in the GP. In addition to the effects of MnB and SF, the welding technique had no further influence on R1. MnB and SF were significant predictors of R1 but not of R2*, indicative of metal accumulation, especially in the GP. Also, high airborne Mn concentration was associated with higher R1 signals in this brain region. The negative results obtained for being a welder or for the techniques with higher exposure to ultrafine particles when the blood

  9. Neutron field characterization in the installation for BNCT study in the IEA-R1 reactor; Caracterizacao do campo de neutrons na instalacao para estudo em BNCT no reator IEA-R1

    Carneiro Junior, Valdeci


    This work aims to characterize the mixed neutron and gamma field, in the sample irradiation position, in a research installation for Boron Neutron Capture Therapy (BNCT), in the IPEN IEA-R1 reactor. The BNCT technique has been studied as a safe and selective option in the treatment of resistant cancerigenous tumors or considered non-curable by the conventional techniques, for example, the Glioblastoma Multiform - a brain cancerigenous tumor. Neutron flux measurements were carried out: thermal, resonance and fast, as well as neutron and gamma rays doses, in the sample position, using activation foils detectors and thermoluminescent dosimeters. For the determination of the neutron spectrum and intensity, a set of different threshold activation foils and gold foils covered and uncovered with cadmium irradiated in the installation was used, analyzed by a high Pure Germanium semiconductor detector, coupled to an electronic system suitable for gamma spectrometry. The results were processed with the SAND-BP code. The doses due to gamma and neutron rays were determined using thermoluminescent dosimeters TLD 400 and TLD 700 sensitive to gamma and TLD 600, sensitive to neutrons. The TLDs were selected and used for obtaining the calibration curves - dosimeter answer versus dose - from each of the TLD three types, which were necessary to calculate the doses due to neutron and gamma, in the sample position. The radiation field, in the sample irradiation position, was characterized flux for thermal neutrons of 1.39.10{sup 8} {+-} 0,12.10{sup 8} n/cm{sup 2}s the doses due to thermal neutrons are three times higher than those due to gamma radiation and confirm the reproducibility and consistency of the experimental findings obtained. Considering these results, the neutron field and gamma radiation showed to be appropriated for research in BNCT. (author)

  10. 2R1T并联支撑机构动基座的自动调平系统设计%Design for the Automatic Leveling System of Parallel Support Mechanism Based on 2R1T

    江涛; 朱大昌


      根据动基座自动调平系统调平特性,采用螺旋理论机构综合方法,给出一类适合作为动基座自动调平系统支撑机构的三自由度(2R1T)并联机构构型。在此基础上,选取一种自动调平系统的支撑机构设计方案,进行了正、逆运动学分析,最后建立了调平支撑机构的三维模型,应用ADAMS软件研究了此种调平机构的动态特性是否满足调平系统的调平特性。%Based on the screw theory,the paper presents a systematic method for structural synthesis of the two translations and one rotation parallel robot.According to the reciprocal product between kinetic screw and constrainted screw in screw theory,this method firstly creats possible branch structures and then generates different models of mechanism.By this method,the paper carries on the structural synthesis of the two translations and one rotation parallel robot.Using software of ADAMS to analyze the mechanism kinematics and dynamics,it analyzes the kinetic characteristics to verify whether it can meet the automatic leveling system dynamics and kinematics of the requirements or not.

  11. Colitogenic role of tumour necrosis factor (TNF) receptors in trinitrobenzene sulphonic acid colitis: TNF-R1 ablation does not affect systemic inflammatory response.

    Yang, Y; Wang, H; Dou, Y; Wang, Y; Han, G; Wang, Renxi; Wang, L; Guo, R; Xiao, H; Li, X; Shen, B; Shi, Y; Chen, G; Li, Y


    Tumour necrosis factor (TNF)-α plays a critical role in the pathogenesis of T helper type 1-mediated colitis such as Crohn's disease. However, the roles of its two receptors in mediating pathology remain largely unknown. In this study, trinitrobenzene sulphonic acid (TNBS) was used to induce colitis in TNF-receptor single or double knock-out (DKO) BALB/c mice and in wild-type counterparts. TNF-R1(-/-) mice had significantly less weight loss, reduced mortality, colon shortening and oedema, colon histological damage and lower levels of colon myeloperoxidase compared with wild-type (WT) BALB/c mice. A similar manifestation was also observed in TNF-R2(-/-) and TNF-R1(-/-) TNF-R2(-/-) (TNF-R DKO) mice. Strikingly, systemic inflammatory response (including splenomegaly and monocyte expansion) was found in WT and TNF-R1(-/-) mice after TNBS, instead of TNF-R2(-/-) and TNF-R DKO mice. Attenuated pathology of colitis in TNF-R1(-/-) or TNF-R2(-/-) mice correlated with lower amounts of interleukin (IL)-6, IL-1β, monocyte chemotactic protein (MCP)-1, IL-12p70 and interferon (IFN)-γ production in the colons. Importantly, ablation of TNF-R1 or TNF-R2 reduced the number of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labelling (TUNEL)-positive apoptotic epithelial cells in the affected colons compared with WT TNBS-instilled controls, which might be due to the heightened ratio of Bcl-2/Bax and reduced activity of nuclear factor (NF)-κB. These findings suggest that either TNF-R1 or TNF-R2 plays a pathogenic role in the pathology of colitis and TNF signalling via TNF-R1 or TNF-R2 alone is not sufficient for inducing mucosal damage.

  12. Monocytes, microglia, and CD200-CD200R1 signaling are essential in the transmission of inflammation from the periphery to the central nervous system.

    Xie, Xin; Luo, Xiaoguang; Liu, Na; Li, Xiaohong; Lou, Fan; Zheng, Yumin; Ren, Yan


    Peripheral inflammation is known to trigger neuroinflammation and neurodegenerative disease. However, the key components during the propagation of inflammation from the periphery to the central nervous system (CNS) remain unclear. Lipopolysaccharide (LPS) was administered to Sprague-Dawley rats to induce peripheral inflammation. An intravenous injection and an intranigral injection of clodronate liposomes were given to deplete monocytes and microglia, respectively. Recombinant CD200 fusion protein (CD200Fc) or an anti-CD200R1 antibody was injected into the substantia nigra to manipulate the involvement of CD200 and CD200R1. Immunohistochemistry and immunofluorescence staining were used to measure microglial activation and dopaminergic neuronal loss. The expression of brain pro-inflammatory cytokines (i.e., tumor necrosis factor alpha, IL-1β) and CD200-CD200R1 signaling were measured by quantitative RT-PCR. Our data showed that the peripheral LPS injection activated the microglia and induced an increase in the levels of pro-inflammatory cytokines (i.e., tumor necrosis factor alpha, IL-1β). The depletion of either monocytes or microglia suppressed these inflammatory effects that were induced by peripheral LPS administration. The peripheral LPS injection increased the expression of CD200 and CD200R1 in the substantia nigra. Dopaminergic neuronal loss induced by the peripheral LPS injection was accelerated by the blockade of CD200-CD200R1 signaling with an anti-CD200R1 antibody and attenuated by intensifying the signaling with CD200Fc. These results highlight the importance of monocytes, microglia, and CD200-CD200R1 signaling in the transmission of inflammation from the periphery to the CNS. © 2017 International Society for Neurochemistry.

  13. 中药复方对戊四氮致痫大鼠海马mGLuR1表达的影响



    目的:观察戊四氮(PTZ)致痫大鼠海马神经元代谢型谷氨酸受体(mGluR1)表达以及中药复方AAP的脑保护作用.方法:动物随机分为6组,复制戊四氮致痫大鼠模型;于致痫后12 h、2 d、5 d、7 d相应时间点取材,制备脑标本;免疫组化技术检测大鼠海马神经元mGluR1表达.结果:与正常组比较,模型组mGluR1免疫反应阳性表达增加,差异显著(P<0.05);与模型组及丙戊酸钠组比较,中药复方AAPl、AAPm、AAPs组海马CA3区mGluR1阳性表达水平降低,差异显著(P<0.05).结论:戊四氮致痫大鼠海马mGluR1表达增加,mGluR1可能在PTZ致大鼠癫痫发作中起作用;中药复方AAP可降低mGluR1表达,对癫痫大鼠有脑保护作用.

  14. Effect of Xiaoer Fuxie Waifu powder on the ultrastructure of intestinal Cajal cells and expression of neurotransmitter receptor VIP-R1 in rats with diarrhea

    Si-wei CHEN


    Full Text Available Objective To investigate the effect of Xiaoer Fuxie Waifu powder on intestinal Cajal cells (ICC and the expression of vasoactive peptide receptor 1 (VIP-R1 in rats with diarrhea. Methods Thirty Wistar rats were divided randomly into three groups: control group, model group, and treatment group (10 each. Folium sennae (2ml/100g was gavaged to reproduce the diarrheal model. Xiaoer Fuxie Waifu powder was applied topically in the treatment group. Transmission electron microscope was used to observe the changes in ultrastructure of ICC after application of the drug. Immunohistochemistry was used to observe the distribution of VIP-R1 in the intestine of diarrheic rats. RT-PCR and Western blotting were used to determine the expressions of mRNA and protein of VIP-R1. Results The ultrastructure of ICC showed that the drug treatment could normalize the cellular morphology and gap junction. VIP-R1 was found to be distributed mainly between circular muscle and longitudinal muscle, in the myenteric nerve plexus of the small intestine, and was found around the epithelial cells in the mucosal layer of the colon and lamina propria cells. Xiaoer Fuxie Waifu powder inhibited the secretion of VIP-R1 and down-regulated the mRNA and protein expression of VIP-R1. Conclusion Xiaoer Fuxie Waifu powder can repair the injured ICC of diarrheic rats and cure diarrhea by down-regulating the expression of VIP-R1. DOI: 10.11855/j.issn.0577-7402.2015.01.05

  15. The awareness of the functional and near population with the relation to the research nuclear reactor IEA-R1

    Vanni, Silvia R.; Martins, Maria da Penha S. [Centro Tecnologico da Marinha (CTMSP), SP (Brazil); Sabundjian, Gaiane, E-mail: gdjian@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    After the natural accident that hit Japan in the beginning of March of 2011, and that ended into an accident of great proportions in the nuclear installations of Fukushima, it has now the debate over the lack of information that the population in general has over the nuclear energy. The dissemination of information, about the operation and security of the nuclear reactors, has the purpose of softening the effect that the pessimistic atmosphere has over its using. This study was reinforced by the memories of serious consequences due to other nuclear accidents that have already happened (Chernobyl, Three-Mile and Hiroshima/Nagasaki event), bringing insecurity, fear and even revenge from part of the public. Over all, people are not sufficiently informed about the positives and negatives aspects of the nuclear energy. It is necessary the adoption of a clear and aware policy with the population, about the pacific use of nuclear energy. Today, the international and national organizations of control of nuclear energy, the International Atomic Energy Agency (IAEA) and the Comissao Nacional de Energia Nuclear (CNEN), have respectively, published information about this subject using a more professional way and of hard access for the public in general. This work has the goal of checking the level of information that the population of workers and individuals of the close public to the research nuclear reactor IEA-R1, located in the Institute of Nuclear Research (IPEN), University City, Sao Paulo, Brazil, has over it. The way used for this study, involved questionnaires with straight questions and of simple language over the subject, to people of all different social, economic and cultural classes, from 12 to 80 years old. From the results found after this work, it was verified the necessity to elaborate a project of awareness of information and clarification about the nuclear energy, using ways of communication that exist and that are easy for the public to understand. (author)

  16. TRAPPIST photometry and imaging monitoring of comet C/2013 R1 (Lovejoy): Implications for the origin of daughter species

    Opitom, C.; Jehin, E.; Manfroid, J.; Hutsemékers, D.; Gillon, M.; Magain, P.


    We report the results of the narrow-band photometry and imaging monitoring of comet C/2013 R1 (Lovejoy) with the robotic telescope TRAPPIST (La Silla observatory). We gathered around 400 images over 8 months pre- and post-perihelion between September 12, 2013 and July 6, 2014. We followed the evolution of the OH, NH, CN, C3, and C2 production rates computed with the Haser model, as well as the evolution of the dust production. All five gas species display an asymmetry about perihelion, since the rate of brightening is steeper than the rate of fading. The study of the coma morphology reveals gas and dust jets that indicate one or several active zone(s) on the nucleus. The dust, C2, and C3 morphologies present some similarities, while the CN morphology is different. OH and NH are enhanced in the tail direction. The study of the evolution of the comet activity shows that the OH, NH, and C2 production rate evolution with the heliocentric distance is correlated to the dust evolution. The CN and, to a lesser extent, the C3 do not display such a correlation with the dust. This evidence and the comparison with parent species production rates indicate that C2 and C3, on one hand, and OH and NH, on the other, could be - at least partially - released from organic - rich grains and icy grains. On the contrary, all evidences point to HCN being the main parent of CN in this comet. Appendix A is available in electronic form at

  17. 1α,25-dihydroxyvitamin D3 modulates CYP2R1 gene expression in human oral squamous cell carcinoma tumor cells.

    Sundaram, Kumaran; Sambandam, Yuvaraj; Tsuruga, Eichi; Wagner, Carol L; Reddy, Sakamuri V


    Oral squamous cell carcinomas (OSCC) are the most common malignant neoplasms associated with mucosal surfaces of the oral cavity and oropharynx. 1α,25-Dihydroxyvitamin D3 (1,25(OH)2D3) is implicated as an anticancer agent. Cytochrome P450 2R1 (CYP2R1) is a microsomal vitamin D 25-hydroxylase which plays an important role in converting dietary vitamin D to active metabolite, 25-(OH)D3. We identified high levels of CYP2R1 expression using tissue microarray of human OSCC tumor specimens compared to normal adjacent tissue. Therefore, we hypothesize that 1,25(OH)2D3 regulates CYP2R1 gene expression in OSCC tumor cells. Interestingly, real-time RT-PCR analysis of total RNA isolated from OSCC cells (SCC1, SCC11B, and SCC14a) treated with 1,25(OH)2D3 showed a significant increase in CYP2R1 and vitamin D receptor (VDR) mRNA expression. Also, Western blot analysis demonstrated that 1,25(OH)2D3 treatment time-dependently increased CYP2R1 expression in these cells. 1,25(OH)2D3 stimulation of OSCC cells transiently transfected with the hCYP2R1 promoter (-2 kb)-luciferase reporter plasmid demonstrated a 4.3-fold increase in promoter activity. In addition, 1,25(OH)2D3 significantly increased c-Fos, p-c-Jun expression, and c-Jun N-terminal kinase (JNK) activity in these cells. The JNK inhibitor suppresses 1,25(OH)2D3, inducing CYP2R1 mRNA expression and gene promoter activity in OSCC cells. Furthermore, JNK inhibitor significantly decreased 1,25(OH)2D3 inhibition of OSCC tumor cell proliferation. Taken together, our results suggest that AP-1 is a downstream effector of 1,25(OH)2D3 signaling to modulate CYP2R1 gene expression in OSCC tumor cells, and vitamin D analogs could be potential therapeutic agents to control OSCC tumor progression.

  18. Tumor necrosis factor-α regulates glucocorticoid synthesis in the adrenal glands of Trypanosoma cruzi acutely-infected mice. the role of TNF-R1.

    Silvina R Villar

    Full Text Available Adrenal steroidogenesis is under a complex regulation involving extrinsic and intrinsic adrenal factors. TNF-α is an inflammatory cytokine produced in response to tissue injury and several other stimuli. We have previously demonstrated that TNF-R1 knockout (TNF-R1(-/- mice have a dysregulated synthesis of glucocorticoids (GCs during Trypanosoma cruzi acute infection. Since TNF-α may influence GCs production, not only through the hypothalamus-pituitary axis, but also at the adrenal level, we now investigated the role of this cytokine on the adrenal GCs production. Wild type (WT and TNF-R1(-/- mice undergoing acute infection (Tc-WT and Tc-TNF-R1(-/- groups, displayed adrenal hyperplasia together with increased GCs levels. Notably, systemic ACTH remained unchanged in Tc-WT and Tc-TNF-R1(-/- compared with uninfected mice, suggesting some degree of ACTH-independence of GCs synthesis. TNF-α expression was increased within the adrenal gland from both infected mouse groups, with Tc-WT mice showing an augmented TNF-R1 expression. Tc-WT mice showed increased levels of P-p38 and P-ERK compared to uninfected WT animals, whereas Tc-TNF-R1(-/- mice had increased p38 and JNK phosphorylation respect to Tc-WT mice. Strikingly, adrenal NF-κB and AP-1 activation during infection was blunted in Tc-TNF-R1(-/- mice. The accumulation of mRNAs for steroidogenic acute regulatory protein and cytochrome P450 were significantly increased in both Tc-WT and Tc-TNF-R1(-/- mice; being much more augmented in the latter group, which also had remarkably increased GCs levels. TNF-α emerges as a potent modulator of steroidogenesis in adrenocortical cells during T. cruzi infection in which MAPK pathways, NF-κB and AP-1 seem to play a role in the adrenal synthesis of pro-inflammatory cytokines and enzymes regulating GCs synthesis. These results suggest the existence of an intrinsic immune-adrenal interaction involved in the dysregulated synthesis of GCs during murine Chagas

  19. Identification of novel regulatory GRE-binding elements in the porcine IP3R1 gene promoter and their transcriptional activation under glucocorticoid stimulation.

    Chai, Jin; Xiong, Qi; Wang, Dan; Wan, Xuebing; Niu, Hongdan; Xiang, Hong; Li, Huanan; Wang, Hongshuai; Zheng, Rong; Peng, Jian; Jiang, Siwen


    Inositol 1,4,5-trisphosphate receptor 1 (IP3R1) is a type of ligand-gated calcium channel that is expressed predominantly in mammalian skeletal muscle, where it acts as a key regulator of calcium homeostasis. In meat, calcium disequilibrium is accompanied by the deterioration of meat quality. Here we show that serum cortisol concentration was higher and the IP3R1 gene expression level increased markedly in pigs exposed to high stress. In porcine primary muscle cells, dexamethasone (DEX, a synthetic glucocorticoid) increased the protein levels of porcine IP3R1 and GRα, and cell apoptosis, and the specific GRα inhibitor RU486 attenuated these effects. DEX also increased the expression of IP3R1 at both the gene and protein levels, and this expression was attenuated by RU486, siRNA against GRα, and the transcriptional inhibitor actinomycin D. DEX significantly reduced cell viability and increased the intracellular calcium concentration, and these effects were attenuated by siRNA against GRα. Bioinformatics analyses predicted a potential glucocorticoid response element (GRE) located in the region -326 to -309 upstream of the IP3R1 promoter and highly conserved in pigs and other mammalian species. Promoter analysis showed that this region containing the GRE was critical for transcriptional activity of porcine IP3R1 under DEX stimulation. This was confirmed by deletion and site-mutation methods. EMSA and ChIP assays showed that this potential GRE bound specifically to GRα and this complex activated the transcription of the IP3R1 gene. Taken together, these data suggest that DEX-mediated induction of IP3R1 influences porcine muscle cells through the transcriptional activation of a mechanism involving interactions between GRα and a GRE present in the proximal IP3R1 promoter. This process can lead to an imbalance in intracellular calcium concentration, which may subsequently activate the apoptosis signal and decrease cell activity, and cause deterioration of meat

  20. Pressure-Dependent Base-Wavefunction Admixture and Lifetime of R1 State of La3Lu2Ga3O12:Cr3+

    ZHANG Zheng-Jie; MA Dong-Ping


    As a key factor leading to the pressure-dependent R1-line-shift reversal and R1-state lifetime, at 10 K,the pressure-dependent variation of mixing-degree of |t22(3T1)e4T2> and |t322E> base-wavefunctions in the wavefunction of R1 state of LLGG:Cr3+ has been calculated and analyzed. From this, the physical origin of the pressure-dependent R1-line-shift reversal has been revealed. Furthermore, by using the pressure-dependent values of the sum of all square mixing-coefficients of|t22 (3T1)e4T2> in the wavefunction of R1 state, the lifetimes of Ri state of LLGG:Cr3+ at various pressures have been calculated, which are in good agreement with observed results. The quantum anticrossing effect between t322E and t22(3T1)e4T2 levels due to both spin-orbital interaction and electron-phonon interaction is remarkable,which is related to the admixture of |t22(3T1)e4T2> and |t32E> as well as the low-high crystal-field transition.

  1. Mutual amplification of HNF4α and IL-1R1 composes an inflammatory circuit in Helicobacter pylori associated gastric carcinogenesis

    Ma, Lin; Zeng, Jiping; Guo, Qing; Liang, Xiuming; Shen, Li; Li, Shuyan; Sun, Yundong; Li, Wenjuan; Liu, Shili; Yu, Han; Chen, Chunyan; Jia, Jihui


    Helicobacter pylori (Hp) is an environmental inducer of gastritis and gastric cancer (GC). The immune response to Hp and the associated changes in somatic gene expression are key determinants governing the transition from gastritis to GC. We show that hepatocyte nuclear factor 4α (HNF4α) is upregulated by Hp infection via NF-κB signaling and that its protein and mRNA levels are elevated in GC. HNF4α in turn stimulates expression of interleukin-1 receptor 1(IL-1R1), which amplifies the inflammatory response evoked by its ligand IL-1β. IL-1β/IL-1R1 activates NF-κB signaling, thereby increasing HNF4α expression and forming a feedback loop that sustains activation of the NF-κB pathway and drives the inflammation towards GC. Examination of clinical samples revealed that HNF4α and IL-1R1 levels increase with increasing severity of Hp-induced gastritis and reach their highest levels in GC. Co-expression of HNF4α and IL-1R1 was a crucial indicator of malignant transformation from gastritis to GC, and was associated with a poorer prognosis in GC patients. Disruption of the HNF4α/IL-1R1/IL-1β/NF-κB circuit during Hp infection maybe an effective means of preventing the associated GC. PMID:26870992

  2. The evolution of doses in the IEA-R1 reactor environment and tendencies based on the current results; Evolucao das doses no ambiente do Reator IEA-R1 e tendencias com base nos resultados atuais

    Toyoda, Eduardo Yoshio


    The IPEN / CNEN-SP have a Nuclear Research Reactor-NRR named IEA-R1, in operation from 1957. It is an open swimming pool reactor using light water as shielding, moderator and as cooling, the volume of this pool is 273m{sup 3}.Until 1995 the reactor operated daily at a power of 2,0 MW. From June of that year, after a few safety modifications the reactor began operating in continuous way from Monday to Wednesday without shutdown totalizing 64 hours per week and the power was increased to 4,5MW also. Because of these changes, continuous operation and increased power, workers' doses would tend to increase. In the past several studies were conducted seeking ways to reduce the workers' doses. A study was made on the possibility to introduce a shielding at the top of the reactor core with a hot water layer. Studies have shown that a major limitation for operating a reactor at high power comes from the gamma radiation emitted by the sodium-24. Other elements such as magnesium-27, aluminum-28, Argon-51, contribute considerably to the water activity of the pool. The introduction of a hot water layer on the swimming pool would form a layer of surface, stable and free of radioactive elements with a 1.5m to 2m thickness creates a shielding to radiation from radioactive elements dissolved in water. Optimization studies proved that the installation of the hot layer was not necessary for the regime and the current power reactor operation, because other procedures adopted were more effective. From this decision the Radiological Protection Reactor Team, set up a dose assessment program to ensure them remained in low values based on principles established in national and international standards. The purpose of this paper is to analyze the individual doses of OEI (Occupationally Exposed Individual), which will be checked increasing doses resulting from recent changes in reactor operation regime and suggested viable safety and protection options, in the first instance to

  3. The FiR 1 photon beam model adjustment according to in-air spectrum measurements with the Mg(Ar) ionization chamber.

    Koivunoro, H; Schmitz, T; Hippeläinen, E; Liu, Y-H; Serén, T; Kotiluoto, P; Auterinen, I; Savolainen, S


    The mixed neutron-photon beam of FiR 1 reactor is used for boron-neutron capture therapy (BNCT) in Finland. A beam model has been defined for patient treatment planning and dosimetric calculations. The neutron beam model has been validated with an activation foil measurements. The photon beam model has not been thoroughly validated against measurements, due to the fact that the beam photon dose rate is low, at most only 2% of the total weighted patient dose at FiR 1. However, improvement of the photon dose detection accuracy is worthwhile, since the beam photon dose is of concern in the beam dosimetry. In this study, we have performed ionization chamber measurements with multiple build-up caps of different thickness to adjust the calculated photon spectrum of a FiR 1 beam model.

  4. Characterization of AvaR1, an autoregulator receptor that negatively controls avermectins production in a high avermectin-producing strain.

    Wang, Jian-Bo; Zhang, Feng; Pu, Jin-Yue; Zhao, Juan; Zhao, Qun-Fei; Tang, Gong-Li


    Many γ-butyrolactone-autoregulator receptors control the production of secondary metabolites in Streptomyces spp. Hence, AvaR1, an autoregulator receptor protein in Streptomyces avermitilis, was characterized as a negative regulator of avermectin (Ave) production. Deletion of AvaR1 in a high-producing strain increased production of Ave B1a approx. 1.75 times (~700 μg/ml) compared with the parent strain. Semi-quantitative RT-PCR and electrophoretic mobility shift assays revealed that AvaR1 regulates the biosynthesis of Ave but not through the aveR pathway-specific regulatory gene. A special signaling molecule, avenolide, increased production of Ave. This study has refined our understanding of how avenolide regulates the production of Aves which is promising for developing new methods to improve the production of antibiotics in industrial strains.

  5. Lack of any prognostic relationship between adiponectin receptor (Adipo R1/R2) expression for early/advanced stage gastric cancer.

    Ayyildiz, Talat; Dolar, Enver; Ugras, Nesrin; Dizdar, Oguzhan Sitki; Adim, Saduman Balaban; Yerci, Omer


    Adiponectin (ApN) is a complement C1q-related protein, mainly secreted from adipose tissue, that signals through ApN receptor 1 (Adipo-R1) and ApN receptor 2 (Adipo-R2). Low serum ApN concentrations are associated with obesity-related malignancies. However, there are very few studies on any prognostic role of ApN receptors in gastric cancer. The aim of this study is to investigate the relationship between AdipoR1/R2 expression and early/advanced stage gastric cancer in terms of clinicopathologic characteristics and survival. Eighteen patients with early and 39 with advanced stage gastric cancer who underwent surgical gastric resection were included in this study. Adipo-R1 expression was low in 2 of the 18 patients with early stage gastric cancer (11.1%), while 4 had low Adipo-R2 expression (22.2%). In those with advanced stage gastric cancer, 7 of 39 had low Adipo-R1 expression (17.9%) and 16 had low Adipo-R2 expression (41%). Adipo-R2 expression was significantly higher (p=0.011) in moderately differentiated tumors when compared to well-differentiated tumors. While there was nearly a statistically significant relationship between TNM stage (T, tumor size; N, regional lymph node; M, whether distant metastases exist) and Adipo-R2 expression (p=0.054), there was no relationship between Adipo-R1/-R2 expression with tumor stage and survival. Adipo-R1/-R2 expression has no prognostic significance of in early/advanced stage gastric cancer.

  6. Acute and chronic cocaine differentially alter the subcellular distribution of AMPA GluR1 subunits in region-specific neurons within the mouse ventral tegmental area

    Lane, D.A.; Jaferi, A.; Kreek, M.J.; Pickel, V.M.


    Cocaine administration increases AMPA GluR1 expression and receptor-mediated activation of the ventral tegmental area (VTA). Functionality is determined, however, by surface availability of these receptors in transmitter- and VTA-region-specific neurons, which may also be affected by cocaine. To test this hypothesis, we used electron microscopic immunolabeling of AMPA GluR1 subunits and tyrosine hydroxylase (TH; the enzyme needed for dopamine synthesis), in the cortical-associated parabrachial (PB) and in the limbic-associated paranigral (PN) VTA of adult male C57BL/6 mice receiving either a single injection (acute) or repeated escalating-doses for 14 days (chronic) of cocaine. Acute cocaine resulted in opposing VTA-region-specific changes in TH-containing dopaminergic dendrites. TH-labeled dendrites within the PB VTA showed increased cytoplasmic GluR1 immunogold particle density consistent with decreased AMPA receptor-mediated glutamatergic transmission. Conversely, TH-labeled dendrites within the PN VTA showed greater surface expression of GluR1 with increases in both synaptic and plasmalemmal GluR1 immunogold density after a single injection of cocaine. These changes diminished in both VTA subregions after chronic cocaine administration. In contrast, non-TH-containing (presumably GABAergic) dendrites showed VTA-region-specific changes only after repeated cocaine administration such that synaptic GluR1 decreased in the PB, but increased in the PN VTA. Taken together, these findings provide ultrastructural evidence suggesting that chronic cocaine not only reverses the respective depression and facilitation of mesocortical (PB) and mesolimbic (PN) dopaminergic neurons elicited by acute cocaine, but also differentially affects synaptic availability of these receptors in non-dopaminergic neurons of each region. These adaptations may contribute to increased cocaine seeking/relapse and decreased reward that is reported with chronic cocaine use. PMID:20553819

  7. Distinct roles for miR-1 and miR-133a in the proliferation and differentiation of rhabdomyosarcoma cells.

    Rao, Prakash K; Missiaglia, Edoardo; Shields, Lauren; Hyde, Greg; Yuan, Bingbing; Shepherd, Christopher J; Shipley, Janet; Lodish, Harvey F


    Rhabdomyosarcoma is the most common soft tissue sarcoma in the pediatric population. As this tumor has an undifferentiated myogenic phenotype, agents that promote differentiation hold particular promise as part of a novel therapeutic approach to combat this type of cancer. In this report, we focus on the contribution of two microRNAs (miRNAs) in rhabdomyosarcomas. Levels of miR-1 and miR-133a are drastically reduced in representative cell lines from each major rhabdomyosarcoma subtype (embryonal and alveolar). Introduction of miR-1 and miR-133a into an embryonal rhabdomyosarcoma-derived cell line is cytostatic, thereby suggesting a tumor suppressor-like role for these myogenic miRNAs. Transcriptional profiling of cells after miR-1 and miR-133a expression reveals that miR-1 (but not miR-133a) exerts a strong promyogenic influence on these poorly differentiated tumor cells. We identify mRNAs that are down-regulated by these miRNAs and propose roles for miR-1 and miR-133a in repressing isoforms of genes that are normally not expressed in muscle. Finally, we show that mRNA targets of miR-1 and miR-133a are up-regulated in rhabdomyosarcomas, suggesting a causative role for these miRNAs in the development of rhabdomyosarcomas. More important, these results point to the promise of enhancing rhabdomyosarcoma therapy using miRNAs as agents that mediate cytostasis and promote muscle differentiation.

  8. Metabotropic glutamate receptor I (mGluR1) antagonism impairs cocaine-induced conditioned place preference via inhibition of protein synthesis.

    Yu, Fei; Zhong, Peng; Liu, Xiaojie; Sun, Dalong; Gao, Hai-Qing; Liu, Qing-Song


    Antagonism of group I metabotropic glutamate receptors (mGluR1 and mGluR5) reduces behavioral effects of drugs of abuse, including cocaine. However, the underlying mechanisms remain poorly understood. Activation of mGluR5 increases protein synthesis at synapses. Although mGluR5-induced excessive protein synthesis has been implicated in the pathology of fragile X syndrome, it remains unknown whether group I mGluR-mediated protein synthesis is involved in any behavioral effects of drugs of abuse. We report that group I mGluR agonist DHPG induced more pronounced initial depression of inhibitory postsynaptic currents (IPSCs) followed by modest long-term depression (I-LTD) in dopamine neurons of rat ventral tegmental area (VTA) through the activation of mGluR1. The early component of DHPG-induced depression of IPSCs was mediated by the cannabinoid CB1 receptors, while DHPG-induced I-LTD was dependent on protein synthesis. Western blotting analysis indicates that mGluR1 was coupled to extracellular signal-regulated kinase (ERK) and mammalian target of rapamycin (mTOR) signaling pathways to increase translation. We also show that cocaine conditioning activated translation machinery in the VTA via an mGluR1-dependent mechanism. Furthermore, intra-VTA microinjections of mGluR1 antagonist JNJ16259685 and protein synthesis inhibitor cycloheximide significantly attenuated or blocked the acquisition of cocaine-induced conditioned place preference (CPP) and activation of translation elongation factors. Taken together, these results suggest that mGluR1 antagonism inhibits de novo protein synthesis; this effect may block the formation of cocaine-cue associations and thus provide a mechanism for the reduction in CPP to cocaine.

  9. Reduced Dendritic Cells Expressing CD200R1 in Children with Inflammatory Bowel Disease: Correlation with Th17 and Regulatory T Cells.

    Elshal, Mohamed F; Aldahlawi, Alia M; Saadah, Omar I; McCoy, J Philip


    Loss of tolerance of the adaptive immune system towards indigenous flora contributes to the development of inflammatory bowel diseases (IBD). Defects in dendritic cell (DC)-mediated innate and adoptive immune responses are conceivable. The aim of this study was to investigate the expression of the inhibitory molecules CD200R1 and their ligand CD200 on DCs, to clarify the role of the DCs in the pathogenesis of IBD. Thirty-seven pediatric IBD patients (23 with Crohn's disease (CD) and 14 with ulcerative colitis (UC)) with mean age 13.25 ± 2.9 years were included. Fourteen age-matched healthy pediatric volunteers (five males and nine females) served as a control group (HC). The percentage of CD11c⁺ myeloid dendritic cells (mDCs) and CD123⁺ plasmacytoid DCs (pDCs) expressing CD200R1 and CD200 were evaluated in peripheral blood using flow cytometry and were correlated with routine biochemical, serological markers, serum levels of cytokines and with the percentages of circulating regulatory T cells (Treg) and CD4⁺ producing IL-17 (Th17). IBD patients showed a significant decrease in the percentage of pDCs and mDCs expressing CD200R1 compared to that of HC. Patients with UC showed increased expressions of the CD200 molecule on pDCs as compared to HC. DCs expressing CD200R1 were found to be correlated positively with Treg and negatively with TH17 and erythrocyte sedimentation rate (ESR). Our findings suggest that IBD is associated with dysregulation in the CD200R1/CD200 axis and that the decrease in DCs expressing CD200R1 may contribute to the imbalance of Th17 and Treg cells and in the pathogenesis of IBD.

  10. MiR-181b regulates cisplatin chemosensitivity and metastasis by targeting TGFβR1/Smad signaling pathway in NSCLC.

    Wang, Xiaoyuan; Chen, Xuesong; Meng, Qingwei; Jing, Hu; Lu, Hailing; Yang, Yanmei; Cai, Li; Zhao, Yanbin


    MicroRNAs (miRNAs) have been identified as important post-transcriptional regulators involved in various biological and pathological processes of cells, but their underlying mechanisms in chemosensitivity and metastasis have not been fully elucidated. The objective of this study was to identify miR-181b and its mechanism in the chemosensitivity and metastasis of NSCLC. We found that miR-181b expression levels were lower in A549/DDP cells compared with A549 cells. Functional assays showed that the overexpression of miR-181b inhibited proliferation, enhanced chemosensitivity to DDP, attenuated migration and metastatic ability in NSCLC cell lines in vitro and in vivo. TGFβR1 was subsequently identified as a novel functional target of miR-181b. TGFβR1 knockdown revealed similar effects as that of ectopic miR-181b expression, whereas overexpression of TGFβR1 rescued the function of miR-181b-mediated growth, chemosensitivity and metastasis in NSCLC cells. In addition, miR-181b could inactivate the TGFβR1/Smad signaling pathway. We also observed that decreased miR-181b expression and increased TGFβR1 expression were significantly associated with chemosensitivity to DDP and tumor metastasis in NSCLC patients. Consequently, miR-181b functions as a tumor suppressor and has an important role in proliferation, chemosensitivity to DDP and metastasis of NSCLC by targeting TGFβR1/Smad signaling pathway.

  11. Biological and geological characteristics of the R1 and R2 coral mounds, Rockall Trough, west of Ireland.

    Unnithan, V.; Grehan, A.; van Weering, T.; Olu-Leroy, K.


    The carbonate mounds discovered in the mid-1990s on the Irish Continental Margin are unique. It is not only their size (up to 300 m in height and 2-3 km in diameter), distribution (along the margins of the Porcupine Seabight and Rockall Trough), abundance (> 250 individual mounds) but also their association with deep-water coral species that has generated a great deal of interest in the scientific community. During the past 10 years a number of European Union funded projects concentrated their efforts on studying these deep-sea features. However, there is still a great deal to be learnt regarding mound structure, dynamics and genesis. The basic question why and how carbonate mounds are formed is still largely unanswered. The CARACOLE (CARbonate And COLD water Ecosystems) Cruise in August 2001, was an Irish-French-EU inter-disciplinary co-operation program with participation of ACES, ECOMOUND and GEOMOUND related scientists from Germany, The Netherlands and Belgium to study carbonate mounds and deep-coral reefs in the Porcupine Seabight and Rockall Trough, west of Ireland. The IFREMER led cruise aboard the French Research Vessel Atalante deployed the 'state of the art' remotely operated vehicle, Victor 6000 at a total of 5 mound locations selected on the basis of previous extensive seismic, acoustic and bottom sampling studies, mainly carried out by RV 'Pelagia' of the Royal NIOZ. High-resolution geo-referenced video and digital still photography was used for detailed observation and mapping. This poster presents preliminary results and work in progress from the R1 and R2 Rockall Trough mound sites based on video and bathymetric analysis carried out by the authors in March 2002 at IFREMER. The focus of the analysis was two fold: 1) Biological, encompassing the identification and mapping of coral habitats and associated species, and 2) geological which includes mapping of the morphology and nature (character) of the seabed. From the observations and analysis

  12. Chronic lung injury in the neonatal rat: up-regulation of TGFβ1 and nitration of IGF-R1 by peroxynitrite as likely contributors to impaired alveologenesis.

    Belcastro, Rosetta; Lopez, Lianet; Li, Jun; Masood, Azhar; Tanswell, A Keith


    Postnatal alveolarization is regulated by a number of growth factors, including insulin-like growth factor-I (IGF-I) acting through the insulin-like growth factor receptor-1 (IGF-R1). Exposure of the neonatal rat lung to 60% O2 for 14 days results in impairments of lung cell proliferation, secondary crest formation, and alveologenesis. This lung injury is mediated by peroxynitrite and is prevented by treatment with a peroxynitrite decomposition catalyst. We hypothesized that one of the mechanisms by which peroxynitrite induces lung injury in 60% O2 is through nitration and inactivation of critical growth factors or their receptors. Increased nitration of both IGF-I and IGF-R1 was evident in 60% O2-exposed lungs, which was reversible by concurrent treatment with a peroxynitrite decomposition catalyst. Increased nitration of the IGF-R1 was associated with its reduced activation, as assessed by IGF-R1 phosphotyrosine content. IGF-I displacement binding plots were conducted in vitro using rat fetal lung distal epithelial cells which respond to IGF-I by an increase in DNA synthesis. When IGF-I was nitrated to a degree similar to that observed in vivo there was minimal, if any, effect on IGF-I displacement binding. In contrast, nitrating cell IGF-R1 to a similar degree to that observed in vivo completely prevented specific binding of IGF-I to the IGF-R1, and attenuated an IGF-I-mediated increase in DNA synthesis. Additionally, we hypothesized that peroxynitrite also impairs alveologenesis by being an upstream regulator of the growth inhibitor, TGFβ1. That 60% O2-induced impairment of alveologenesis was mediated in part by TGFβ1 was confirmed by demonstrating an improvement in secondary crest formation when 60% O2-exposed pups received concurrent treatment with the TGFß1 activin receptor-like kinase, SB 431542. That the increased TGFβ1 content in lungs of pups exposed to 60% O2 was regulated by peroxynitrite was confirmed by its attenuation by concurrent treatment

  13. Angiotensin II reduces cardiac AdipoR1 expression through AT1 receptor/ROS/ERK1/2/c-Myc pathway.

    Li Li

    Full Text Available Adiponectin, an abundant adipose tissue-derived protein, exerts protective effect against cardiovascular disease. Adiponectin receptors (AdipoR1 and AdipoR2 mediate the beneficial effects of adiponectin on the cardiovascular system. However, the alteration of AdipoRs in cardiac remodeling is not fully elucidated. Here, we investigated the effect of angiotensin II (AngII on cardiac AdipoRs expression and explored the possible molecular mechanism. AngII infusion into rats induced cardiac hypertrophy, reduced AdipoR1 but not AdipoR2 expression, and attenuated the phosphorylations of adenosine monophosphate-activated protein kinase and acetyl coenzyme A carboxylase, and those effects were all reversed by losartan, an AngII type 1 (AT1 receptor blocker. AngII reduced expression of AdipoR1 mRNA and protein in cultured neonatal rat cardiomyocytes, which was abolished by losartan, but not by PD123319, an AT2 receptor antagonist. The antioxidants including reactive oxygen species (ROS scavenger NAC, NADPH oxidase inhibitor apocynin, Nox2 inhibitor peptide gp91 ds-tat, and mitochondrial electron transport chain complex I inhibitor rotenone attenuated AngII-induced production of ROS and phosphorylation of extracellular signal-regulated kinase (ERK 1/2. AngII-reduced AdipoR1 expression was reversed by pretreatment with NAC, apocynin, gp91 ds-tat, rotenone, and an ERK1/2 inhibitor PD98059. Chromatin immunoprecipitation assay demonstrated that AngII provoked the recruitment of c-Myc onto the promoter region of AdipoR1, which was attenuated by PD98059. Moreover, AngII-induced DNA binding activity of c-Myc was inhibited by losartan, NAC, apocynin, gp91 ds-tat, rotenone, and PD98059. c-Myc small interfering RNA abolished the inhibitory effect of AngII on AdipoR1 expression. Our results suggest that AngII inhibits cardiac AdipoR1 expression in vivo and in vitro and AT1 receptor/ROS/ERK1/2/c-Myc pathway is required for the downregulation of AdipoR1 induced by AngII.

  14. The Production of Benzaldehyde by Rhizopus oligosporus USM R1 in a Solid State Fermentation (SSF) System of Soy Bean Meal: Rice Husks

    Norliza, A. W.; Ibrahim, C. O.


    The cultivation of Rhizopus oligosporus USM R1 for the production of benzaldehyde, a bitter cherry almond flavour was performed using soya bean meal and rice husks as the substrates. The identification of R. oligosporus USM R1 was performed based on the observation made under light microscope and scanning electron microscope (SEM). The optimum conditions for the SSF in a 250-ml Erlenmeyer flask system were 40% (v/w) water content, substrate particle size of 0.7 mm; inoculum size of 1 x 10^5 s...

  15. Mapatumumab, a Fully Human Agonistic Monoclonal Antibody That Targets TRAIL-R1, in Combination with Gemcitabine and Cisplatin : a Phase I Study

    Mom, Constantijne H.; Verweij, Jaap; Oldenhuis, Corina N. A. M.; Gietema, Jourik A.; Fox, Norma Lynn; Miceli, Rene; Eskens, Ferry A. L. M.; Loos, Walter J.; de Vries, Elisabeth G. E.; Sleijfer, Stefan


    Purpose: To evaluate the safety, tolerability, pharmacokinetics, and antitumor activity of mapatumumab, a fully human monoclonal antibody targeting tumor necrosis factor-related apoptosis-inducing ligand receptor 1 (TRAIL-R1), in combination with gemcitabine and cisplatin. Experimental Design:

  16. The architecture of the BubR1 tetratricopeptide tandem repeat defines a protein motif underlying mitotic checkpoint-kinetochore communication.

    Bolanos-Garcia, Victor M; Nilsson, Jakob; Blundell, Tom L


    The accurate and timely transmission of the genetic material to progeny during successive rounds of cell division is sine qua non for the maintenance of genome stability. Eukaryotic cells have evolved a surveillance mechanism, the mitotic spindle assembly checkpoint (SAC), to prevent premature advance to anaphase before every chromosome is properly attached to microtubules of the mitotic spindle. The architecture of the KNL1-BubR1 complex reveals important features of the molecular recognition between SAC components and the kinetochore. The interaction is important for a functional SAC as substitution of BubR1 residues engaged in KNL1 binding impaired the SAC and BubR1 recruitment into checkpoint complexes in stable cell lines. Here we discuss the implications of the disorder-to-order transition of KNL1 upon BubR1 binding for SAC signaling and propose a mechanistic model of how BUBs binding may affect the recognition of KNL1 by its other interacting partners.

  17. Structure-activity relationships of novel non-competitive mGluR1 antagonists: a potential treatment for chronic pain.

    Owen, Dafydd R; Dodd, Peter G; Gayton, Simon; Greener, Ben S; Harbottle, Gareth W; Mantell, Simon J; Maw, Graham N; Osborne, Simon A; Rees, Huw; Ringer, Tracy J; Rodriguez-Lens, Margarita; Smith, Graham F


    A series of novel mGluR1 antagonists have been prepared. Incorporation of fragments derived from weak lead matter into a library led to enhanced potency in a new chemical series. A chemistry driven second library iteration, covering a greatly enhanced area of chemical space, maintained good potency and introduced metabolic stability.

  18. Coupling between the Basic Replicon and the Kis-Kid Maintenance System of Plasmid R1: Modulation by Kis Antitoxin Levels and Involvement in Control of Plasmid Replication

    Juan López-Villarejo


    Full Text Available kis-kid, the auxiliary maintenance system of plasmid R1 and copB, the auxiliary copy number control gene of this plasmid, contribute to increase plasmid replication efficiency in cells with lower than average copy number. It is thought that Kis antitoxin levels decrease in these cells and that this acts as the switch that activates the Kid toxin; activated Kid toxin reduces copB-mRNA levels and this increases RepA levels that increases plasmid copy number. In support of this model we now report that: (i the Kis antitoxin levels do decrease in cells containing a mini-R1 plasmid carrying a repA mutation that reduces plasmid copy number; (ii kid-dependent replication rescue is abolished in cells in which the Kis antitoxin levels or the CopB levels are increased. Unexpectedly we found that this coordination significantly increases both the copy number of the repA mutant and of the wt mini-R1 plasmid. This indicates that the coordination between plasmid replication functions and kis-kid system contributes significantly to control plasmid R1 replication.

  19. Cross-Correlated Relaxation of Dipolar Coupling and Chemical-Shift Anisotropy in Magic-Angle Spinning R1ρ NMR Measurements: Application to Protein Backbone Dynamics Measurements.

    Kurauskas, Vilius; Weber, Emmanuelle; Hessel, Audrey; Ayala, Isabel; Marion, Dominique; Schanda, Paul


    Transverse relaxation rate measurements in magic-angle spinning solid-state nuclear magnetic resonance provide information about molecular motions occurring on nanosecond-to-millisecond (ns-ms) time scales. The measurement of heteronuclear ((13)C, (15)N) relaxation rate constants in the presence of a spin-lock radiofrequency field (R1ρ relaxation) provides access to such motions, and an increasing number of studies involving R1ρ relaxation in proteins have been reported. However, two factors that influence the observed relaxation rate constants have so far been neglected, namely, (1) the role of CSA/dipolar cross-correlated relaxation (CCR) and (2) the impact of fast proton spin flips (i.e., proton spin diffusion and relaxation). We show that CSA/D CCR in R1ρ experiments is measurable and that the CCR rate constant depends on ns-ms motions; it can thus provide insight into dynamics. We find that proton spin diffusion attenuates this CCR due to its decoupling effect on the doublet components. For measurements of dynamics, the use of R1ρ rate constants has practical advantages over the use of CCR rates, and this article reveals factors that have so far been disregarded and which are important for accurate measurements and interpretation.

  20. Telmisartan ameliorates adipoR1 and adipoR2 expression via PPAR-γ activation in the coronary artery and VSMCs.

    Shen, Xuhua; Li, Hongwei; Li, Weiping; Wu, Xing; Sun, Zhijun; Ding, Xiaosong


    The effects of telmisartan on insulin-resistant properties and expression of adiponectin receptors (AdipoRs) were investigated. A diabetic rat model was established using a high-fat diet and streptozotocin (25mg/kg) and primary rat coronary vascular smooth muscle cells (VSMCs) were used to elucidate the underlying mechanisms. The diabetic rats were insulin-resistant and exhibited weight gain, elevated blood pressures, and increased plasma triglyceride levels. These manifestations were ameliorated by elmisartan treatment. Four-week telmisartan therapy increased plasma adiponectin and decreased TNF-α expression in the coronary artery. Moreover, telmisartan significantly decreased AdipoR1 and AdipoR2 expression. Using high glucose-treated rat coronary VSMCs, telmisartan and PPAR-γ agonist GW1929 prominently stimulated PPAR-γ and decreased TNF-α expression. Interestingly, telmisartan or GW1929 also prevented hyperglycemia-induced downregulation of AdipoR1 and AdipoR2 expression. Additionally, GW9662 (PPAR-γ antagonist) significantly decreased the effects of telmisartan on AdipoR1 and AdipoR2 expression. These results demonstrated that telmisartan effectively ameliorated coronary insulin resistance and inflammation in diabetic rats and upregulated AdipoR1/R2 expression via activation of PPAR-γ in the coronary artery and VSMCs. Copyright © 2017. Published by Elsevier Masson SAS.

  1. Galanin-induced decreases in nucleus accumbens/striatum EPSPs and morphine conditioned place preference require both GalR1 and GalR2

    Einstein, Emily B.; Asaka, Yukiko; Yeckel, Mark F.; Higley, Michael J.; Picciotto, Marina R.


    The neuropeptide galanin has been shown to alter the rewarding properties of morphine. To identify potential cellular mechanisms that might be involved in the ability of galanin to modulate opiate reward, we measured excitatory post-synaptic potentials (EPSPs) using both field and whole-cell recordings from striatal brain slices extracted from wild type mice and mice lacking specific galanin receptor (GalR) subtypes. We found that galanin decreases the amplitude of EPSPs in both the dorsal striatum and nucleus accumbens. We then performed recordings in slices from knockout mice lacking either the GalR1 or GalR2 gene and found that the ability of galanin to decrease EPSP amplitude was absent from both mouse lines, suggesting that both receptor subtypes are required for this effect. In order to determine whether behavioral responses to opiates were dependent on the same receptor subtypes, we tested GalR1 and GalR2 mice for morphine conditioned place preference (CPP). Morphine CPP was significantly attenuated in both GalR1 and GalR2 knockout mice. These data suggest that mesolimbic excitatory signaling is significantly modulated by galanin in a GalR1- and GalR2-dependent manner and that morphine CPP is dependent on the same receptor subtypes. PMID:23387435

  2. Crystallization and preliminary X-ray crystallographic studies on the parD-encoded protein Kid from Escherichia coli plasmid R1

    Hargreaves, D.; Giraldo, R.; Santos-Sierra, S.; Boelens, R.; Rice, D.W.; Díaz Orejas, R.; Rafferty, J.B.


    DNA replication in Escherichia coli and therefore bacterial proliferation relies upon the efficient functioning of the DnaB helicase. The toxin protein Kid from the plasmid-stability system parD encoded on plasmid R1 of E. coli is thought to target and block DnaB-dependent DNA replication. The

  3. De novo PIK3R1 gain-of-function with recurrent sinopulmonary infections, long-lasting chronic CMV-lymphadenitis and microcephaly.

    Kuhlen, Michaela; Hönscheid, Andrea; Loizou, Loizos; Nabhani, Schafiq; Fischer, Ute; Stepensky, Polina; Schaper, Jörg; Klapper, Wolfram; Siepermann, Meinolf; Schuster, Friedhelm; Meisel, Roland; Borkhardt, Arndt


    PIK3R1 (phosphoinositide-3-kinase, regulatory subunit 1) gain-of-function has recently been described in patients with recurrent sinopulmonary infections, chronic CMV-/EBV-infections, lymphoproliferation, and hypogammaglobulinemia. Here we report a 15-year-old boy with treatment refractory CMV lymphadenitis, severe combined immunodeficiency, microcephaly and a severe developmental defect of Th17 cells. To avoid poor outcome, hematopoietic stem cell transplantation (HSCT) was performed. Subsequently, whole exome sequencing revealed a de novo heterozygous G-to-C mutation (chr5: 5:67,589,663: G>C) at the splice donor site of the PIK3R1 gene. Our data suggest that PIK3R1 gain-of-function leads to developmental defects in helper and regulatory T-cell subsets, the latter expanding the immunological features of PIK3R1 gain-of-function. T-cell subsets play a critical role in the regulation of immune response against infectious agents and of autoimmunity and thus may be particularly accountable for the clinical phenotype of affected patients. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Chrysin, Apigenin and Acacetin Inhibit Tumor Necrosis Factor-Related Apoptosis—Inducing Ligand Receptor-1 (TRAIL-R1 on Activated RAW264.7 Macrophages

    Monika Warat


    Full Text Available Expression level of Tumor Necrosis Factor—related apoptosis—inducing ligand (TRAIL receptors is one of the most important factors of TRAIL-mediated apoptosis in cancer cells. We here report for the first time data concerning TRAIL-R1 and TRAIL-R2 receptor expression on RAW264.7 macrophages. Three substances belonging to flavones: chrysin, apigenin and acacetin which differ from their substituents at the 4' position in the phenyl ring were used in assays because of the variety of biological activities (e.g., anticancer activity of the polyphenol compounds. The expression of TRAIL-R1 and TRAIL-R2 death receptors on non-stimulated and LPS (lipopolysaccharide-stimulated macrophages was determined using flow cytometry. We demonstrate that RAW264.7 macrophages exhibit TRAIL-R1 surface expression and that the tested compounds: chrysin, apigenin and acacetin can inhibit TRAIL-R1 death receptor expression level on macrophages.

  5. Purification, crystallization and preliminary X-ray diffraction analysis of the IL-20-IL-20R1-IL-20R2 complex

    Logsdon, Naomi J.; Allen, Christopher E.; Rajashankar, Kanagalaghatta R.; Walter, Mark R. (Cornell); (UAB)


    Interleukin-20 (IL-20) is an IL-10-family cytokine that regulates innate and adaptive immunity in skin and other tissues. In addition to protecting the host from various external pathogens, dysregulated IL-20 signaling has been shown to contribute to the pathogenesis of human psoriasis. IL-20 signals through two cell-surface receptor heterodimers, IL-20R1-IL-20R2 and IL-22R1-IL-20R2. In this report, crystals of the IL-20-IL-20R1-IL-20R2 ternary complex have been grown from polyethylene glycol solutions. The crystals belonged to space group P4{sub 1}2{sub 1}2 or P4{sub 3}2{sub 1}2, with unit-cell parameters a = 111, c = 135 {angstrom}, and diffracted X-rays to 3 {angstrom} resolution. The crystallographic asymmetric unit contains one IL-20-IL-20R1-IL-20R2 complex, corresponding to a solvent content of approximately 54%.

  6. International Conference on Harmonisation; guidance on S2(R1) Genotoxicity Testing and Data Interpretation for Pharmaceuticals intended for Human Use; availability. Notice.


    The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled ``S2(R1) Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use'' (ICH S2(R1)). This guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The ICH S2(R1) combines and replaces two ICH guidances, "S2A Specific Aspects for Regulatory Genotoxicity Tests for Pharmaceuticals'' and "S2B Genotoxicity: A Standard Battery for Genotoxicity Testing of Pharmaceuticals.'' ICH S2(R1) provides guidance to drug sponsors on which tests should be performed to assess potential genotoxicity of pharmaceuticals. It also provides guidance on testing conditions, data interpretation, and followup strategies if a positive response is seen in in vitro assays. This guidance is intended to provide drug sponsors with recommendations to ensure that drugs are appropriately tested for potential to cause genetic damage and to ensure efficient development of new drugs.

  7. Crystal Structure of Human Interferon-[lamda]1 in Complex with Its High-Affinity Receptor Interferon-[lamda]R1

    Miknis, Zachary; Magracheva, Eugenia; Li, Wei; Zdanov, Alexander; Kotenko, Sergei V.; Wlodawer, Alexander (NJMS); (NCI)


    Interferon (IFN)-{lambda}1 [also known as interleukin (IL)-29] belongs to the recently discovered group of type III IFNs. All type III IFNs initiate signaling processes through formation of specific heterodimeric receptor complexes consisting of IFN-{lambda}R1 and IL-10R2. We have determined the structure of human IFN-{lambda}1 complexed with human IFN-{lambda}R1, a receptor unique to type III IFNs. The overall structure of IFN-{lambda}1 is topologically similar to the structure of IL-10 and other members of the IL-10 family of cytokines. IFN-{lambda}R1 consists of two distinct domains having fibronectin type III topology. The ligand-receptor interface includes helix A, loop AB, and helix F on the IFN site, as well as loops primarily from the N-terminal domain and inter-domain hinge region of IFN-{lambda}R1. Composition and architecture of the interface that includes only a few direct hydrogen bonds support an idea that long-range ionic interactions between ligand and receptor govern the process of initial recognition of the molecules while hydrophobic interactions finalize it.

  8. Circular dichroism and fluorescence spectroscopy of cysteinyl-tRNA synthetase from Halobacterium salinarum ssp. NRC-1 demonstrates that group I cations are particularly effective in providing structure and stability to this halophilic protein.

    Christopher J Reed

    Full Text Available Proteins from extremophiles have the ability to fold and remain stable in their extreme environment. Here, we investigate the presence of this effect in the cysteinyl-tRNA synthetase from Halobacterium salinarum ssp. NRC-1 (NRC-1, which was used as a model halophilic protein. The effects of salt on the structure and stability of NRC-1 and of E. coli CysRS were investigated through far-UV circular dichroism (CD spectroscopy, fluorescence spectroscopy, and thermal denaturation melts. The CD of NRC-1 CysRS was examined in different group I and group II chloride salts to examine the effects of the metal ions. Potassium was observed to have the strongest effect on NRC-1 CysRS structure, with the other group I salts having reduced strength. The group II salts had little effect on the protein. This suggests that the halophilic adaptations in this protein are mediated by potassium. CD and fluorescence spectra showed structural changes taking place in NRC-1 CysRS over the concentration range of 0-3 M KCl, while the structure of E. coli CysRS was relatively unaffected. Salt was also shown to increase the thermal stability of NRC-1 CysRS since the melt temperature of the CysRS from NRC-1 was increased in the presence of high salt, whereas the E. coli enzyme showed a decrease. By characterizing these interactions, this study not only explains the stability of halophilic proteins in extremes of salt, but also helps us to understand why and how group I salts stabilize proteins in general.

  9. IgE FcɛR1β polymorphism and risk of developing chronic spontaneous urticaria: A study in an ethnic Kashmiri population.

    Rasool, R; Shera, I A; Nissar, S; Yousuf, Q; Shah, Z A


    The pathogenesis of chronic spontaneous urticaria involves interplay between the genetic and environmental factors, most of which is still poorly understood. It is well-recognized that 30-40% of chronic spontaneous urticaria is autoimmune in nature. Chronic autoimmune urticaria is caused by anti-FcɛR1β and less frequently, by anti-IgE auto antibodies that lead to mast cell and basophil activation, thereby giving rise to the release of histamine and other proinflammatory mediators. We investigated the association between SNP loci in FcɛR1β and chronic spontaneous urticaria and to see its relation with serum IgE levels in Kashmiri population. The autologous serum skin test was used as a screening test for chronic autoimmune urticaria. PCR-RFLP was used to detect the genotype of the SNP loci. Serum IgE levels were assessed by ELISA kit. No significant difference was found between the study population and control group in genotype distribution (wild and variant) among FcɛR1β loci (P value=0.06, odds ratio=0.29). The frequency of FcɛR1β (C109T) in autologous serum skin test positive chronic autoimmune urticaria patients with the CT genotype was found to be statistically non-significant when compared with the wild genotype (P=0.35). Carriers of FcɛR1β (T allele) had a more significant risk of developing CAU than those with C allele (P=0.01). In our population serum total IgE levels did not find any statistical significance with regard to ASST positive & ASST negative patients (P=0.26). There is statistically no significant association between FcɛR1β gene polymorphism and CSU in Kashmiri population; however, there is a probability of developing CSU in patients carrying FcɛR1β T allele. Furthermore, serum total IgE levels had no significant association with the development of CAU. Copyright © 2013 SEICAP. Published by Elsevier Espana. All rights reserved.

  10. Automated mass action model space generation and analysis methods for two-reactant combinatorially complex equilibriums: An analysis of ATP-induced ribonucleotide reductase R1 hexamerization data

    Radivoyevitch Tomas


    Full Text Available Abstract Background Ribonucleotide reductase is the main control point of dNTP production. It has two subunits, R1, and R2 or p53R2. R1 has 5 possible catalytic site states (empty or filled with 1 of 4 NDPs, 5 possible s-site states (empty or filled with ATP, dATP, dTTP or dGTP, 3 possible a-site states (empty or filled with ATP or dATP, perhaps two possible h-site states (empty or filled with ATP, and all of this is folded into an R1 monomer-dimer-tetramer-hexamer equilibrium where R1 j-mers can be bound by variable numbers of R2 or p53R2 dimers. Trillions of RNR complexes are possible as a result. The problem is to determine which are needed in models to explain available data. This problem is intractable for 10 reactants, but it can be solved for 2 and is here for R1 and ATP. Results Thousands of ATP-induced R1 hexamerization models with up to three (s, a and h ATP binding sites per R1 subunit were automatically generated via hypotheses that complete dissociation constants are infinite and/or that binary dissociation constants are equal. To limit the model space size, it was assumed that s-sites are always filled in oligomers and never filled in monomers, and to interpret model terms it was assumed that a-sites fill before h-sites. The models were fitted to published dynamic light scattering data. As the lowest Akaike Information Criterion (AIC of the 3-parameter models was greater than the lowest of the 2-parameter models, only models with up to 3 parameters were fitted. Models with sums of squared errors less than twice the minimum were then partitioned into two groups: those that contained no occupied h-site terms (508 models and those that contained at least one (1580 models. Normalized AIC densities of these two groups of models differed significantly in favor of models that did not include an h-site term (Kolmogorov-Smirnov p -15; consistent with this, 28 of the top 30 models (ranked by AICs did not include an h-site term and 28

  11. miR-1/133对病毒性心肌炎小鼠心肌细胞离子通道表达的影响%miR-1/133 participates in the expression of ion channel genes of myocardial cells in myocarditis mice

    刘蒙蒙; 赵翠芬; 孔清玉; 蔡直锋; 夏伟


    目的:探讨 miR-1/133是否参与病毒性心肌炎心肌细胞钾离子、钙离子通道基因表达的调节。方法建立Balb/c 小鼠急性病毒性心肌炎模型。将小鼠分为对照组、心肌炎组、心肌炎+miR-1/133mimics 组、心肌炎+miR-1/133 NC 组,每组10只。采用苏木精-伊红染色法观察心肌形态学改变;采用 qRT-PCR 法检测心肌中 miR-1、miR-133及 Kcnd2、Irx5、Kcnj2和α1c 的相对表达量;采用 Western blotting 法检测心肌中蛋白 Kv4.2、Kir2.1、Cav1.2的相对表达量。结果苏木精-伊红染色显示对照组心肌细胞排列整齐,间质无炎性细胞浸润;心肌炎组与心肌炎+miR-1/133 NC 组心肌细胞水肿、排列紊乱,炎性细胞浸润间质;心肌炎+miR-1/133mimics 组心肌细胞排列较整齐,无细胞水肿,间质少量炎性细胞浸润。与对照组相比,心肌炎组与心肌炎+miR-1/133 NC 组心肌miR-1、miR-133及 Kcnd2、Kcnj2表达下调,蛋白 Kv4.2、Kir2.1的表达下调(P <0.01);Irx5、α1c 及蛋白 Cav1.2表达均上调(P <0.01);心肌炎+miR-1/133 mimics 组较心肌炎组与心肌炎+miR-1/133 NC 组相比,miR-1、miR-133及 Kcnd2、Kcnj2表达上调,蛋白 Kv4.2、Kir2.1表达上调(P <0.05),Irx5、α1c 及蛋白 Cav1.2表达均下调(P <0.01)。结论miR-1/133参与病毒性心肌炎心肌细胞钾离子和钙离子通道基因表达的调节。%Objective To explore the involvement of miR-1 /133 in regulating the expression of potassium and calcium ion channel in mouse with acute viral myocarditis.Methods After the mice models of acute viral myocarditis were es-tablished,they were divided into 4 groups:control group,viral myocarditis group,myocarditis +miR-1 /133 mimics group,and myocarditis +miR-1 /133 NC group,with 10 mice in each group.The myocardial pathological changes were observed with HE staining.The expressions of miR-1,mi

  12. AcvR1-mediated BMP signaling in second heart field is required for arterial pole development: implications for myocardial differentiation and regional identity.

    Thomas, Penny S; Rajderkar, Sudha; Lane, Jamie; Mishina, Yuji; Kaartinen, Vesa


    BMP signaling plays an essential role in second heart field-derived heart and arterial trunk development, including myocardial differentiation, right ventricular growth, and interventricular, outflow tract and aortico-pulmonary septation. It is mediated by a number of different BMP ligands, and receptors, many of which are present simultaneously. The mechanisms by which they regulate morphogenetic events and degree of redundancy amongst them have still to be elucidated. We therefore assessed the role of BMP Type I receptor AcvR1 in anterior second heart field-derived cell development, and compared it with that of BmpR1a. By removing Acvr1 using the driver Mef2c[AHF]-Cre, we show that AcvR1 plays an essential role in arterial pole morphogenesis, identifying defects in outflow tract wall and cushion morphology that preceded a spectrum of septation defects from double outlet right ventricle to common arterial trunk in mutants. Its absence caused dysregulation in gene expression important for myocardial differentiation (Isl1, Fgf8) and regional identity (Tbx2, Tbx3, Tbx20, Tgfb2). Although these defects resemble to some degree those in the equivalent Bmpr1a mutant, a novel gene knock-in model in which Bmpr1a was expressed in the Acvr1 locus only partially restored septation in Acvr1 mutants. These data show that both BmpR1a and AcvR1 are needed for normal heart development, in which they play some non-redundant roles, and refine our understanding of the genetic and morphogenetic processes underlying Bmp-mediated heart development important in human congenital heart disease.

  13. mGluR1/5 subtype-specific calcium signalling and induction of long-term potentiation in rat hippocampal oriens/alveus interneurones

    Topolnik, Lisa; Azzi, Mounia; Morin, France; Kougioumoutzakis, André; Lacaille, Jean-Claude


    Hippocampal inhibitory interneurones demonstrate pathway- and synapse-specific rules of transmission and plasticity, which are key determinants of their role in controlling pyramidal cell excitability. Mechanisms underlying long-term changes at interneurone excitatory synapses, despite their importance, remain largely unknown. We use two-photon calcium imaging and whole-cell recordings to determine the Ca2+ signalling mechanisms linked specifically to group I metabotropic glutamate receptors (mGluR1α and mGluR5) and their role in hebbian long-term potentiation (LTP) in oriens/alveus (O/A) interneurones. We demonstrate that mGluR1α activation elicits dendritic Ca2+ signals resulting from Ca2+ influx via transient receptor potential (TRP) channels and Ca2+ release from intracellular stores. By contrast, mGluR5 activation produces dendritic Ca2+ transients mediated exclusively by intracellular Ca2+ release. Using Western blot analysis and immunocytochemistry, we show mGluR1α-specific extracellular signal-regulated kinase (ERK1/2) activation via Src in CA1 hippocampus and, in particular, in O/A interneurones. Moreover, we find that mGluR1α/TRP Ca2+ signals in interneurone dendrites are dependent on activation of the Src/ERK cascade. Finally, this mGluR1α-specific Ca2+ signalling controls LTP at interneurone synapses since blocking either TRP channels or Src/ERK and intracellular Ca2+ release prevents LTP induction. Thus, our findings uncover a novel molecular mechanism of interneurone-specific Ca2+ signalling, critical in regulating synaptic excitability in hippocampal networks. PMID:16740609

  14. Role of amino-terminal half of the S4-S5 linker in type 1 ryanodine receptor (RyR1) channel gating.

    Murayama, Takashi; Kurebayashi, Nagomi; Oba, Toshiharu; Oyamada, Hideto; Oguchi, Katsuji; Sakurai, Takashi; Ogawa, Yasuo


    The type 1 ryanodine receptor (RyR1) is a Ca(2+) release channel found in the sarcoplasmic reticulum of skeletal muscle and plays a pivotal role in excitation-contraction coupling. The RyR1 channel is activated by a conformational change of the dihydropyridine receptor upon depolarization of the transverse tubule, or by Ca(2+) itself, i.e. Ca(2+)-induced Ca(2+) release (CICR). The molecular events transmitting such signals to the ion gate of the channel are unknown. The S4-S5 linker, a cytosolic loop connecting the S4 and S5 transmembrane segments in six-transmembrane type channels, forms an α-helical structure and mediates signal transmission in a wide variety of channels. To address the role of the S4-S5 linker in RyR1 channel gating, we performed alanine substitution scan of N-terminal half of the putative S4-S5 linker (Thr(4825)-Ser(4829)) that exhibits high helix probability. The mutant RyR1 was expressed in HEK cells, and CICR activity was investigated by caffeine-induced Ca(2+) release, single-channel current recordings, and [(3)H]ryanodine binding. Four mutants (T4825A, I4826A, S4828A, and S4829A) had reduced CICR activity without changing Ca(2+) sensitivity, whereas the L4827A mutant formed a constitutive active channel. T4825I, a disease-associated mutation for malignant hyperthermia, exhibited enhanced CICR activity. An α-helical wheel representation of the N-terminal S4-S5 linker provides a rational explanation to the observed activities of the mutants. These results suggest that N-terminal half of the S4-S5 linker may form an α-helical structure and play an important role in RyR1 channel gating.

  15. SIRT1 attenuates neuropathic pain by epigenetic regulation of mGluR1/5 expressions in type 2 diabetic rats.

    Zhou, Cheng-Hua; Zhang, Ming-Xing; Zhou, Sha-Sha; Li, Huan; Gao, Jian; Du, Lei; Yin, Xiao-Xing


    Accumulating evidence has demonstrated that epigenetic modification-mediated changes in pain-related gene expressions play an important role in the development and maintenance of neuropathic pain. Sirtuin 1 (SIRT1), a nicotinamide adenine dinucleotide (NAD)-dependent deacetylase, is involved in the development of chronic pain. Moreover, SIRT1 may be a novel therapeutic target for the prevention of type 2 diabetes mellitus (T2DM). But the role of SIRT1 in T2DM-induced neuropathic pain remains unknown. In this study, we found that spinal SIRT1 expression and activity were downregulated significantly in high-fat-fed/low-dose streptozotocin-induced neuropathic pain rats. SIRT1 localized in spinal neurons but not in astrocytes or microglia. Furthermore, the expressions of metabotropic glutamate receptor (mGluR1) and mGluR5, which play a key role in central sensitization and neuropathic pain, and H3 acetylation levels at Grm1/5 (encoding mGluR1/5) promoter regions were increased in diabetic neuropathic pain rats. SIRT1 activator SRT1720 reversed thermal hyperalgesia and mechanical allodynia and spinal neuronal activation in diabetic neuropathic pain rats. Concurrently, increased expressions of mGluR1/5 and H3 acetylation levels at Grm1/5 promoter regions were reversed by SIRT1 activation. In addition, knockdown of SIRT1 by Ad-SIRT1-shRNA induced pain behaviors and spinal neuronal activation in normal rats, which was accompanied by the increased expressions of mGluR1/5 and H3 acetylation levels at Grm1/5 promoter regions. Therefore, we concluded that SIRT1-mediated epigenetic regulation of mGluR1/5 expressions was involved in the development of neuropathic pain in type 2 diabetic rats.

  16. Effect of behavior training on learning, memory and the expression of NR2B, GluR1 in hippocampus of rats offspring with fetal growth restriction

    Chunfang Li; Wenli Gou; Yunping Sun; Huang Pu


    Objective: To study effects of behavior training on learning, memory and the expression of NR2B, GIuR1 in hippocampus of rat's offspring with fetal growth restricfion(FGR). Methods: The rat model of FGR was established by passive smoking method. The rats offspring were divided into the FGR group and the control group, then randomly divided into the trained and untrained group, respectively. Morris water maze test was procezded on postnatal month(PM2/4) as a behavior training method, then the learning-memory of rats was detected through dark-avoidance and step-down tests. The expressions of NR2B and GluR1 suhunits in hippocampal CA1 and CA3 areas were detected by immunohistochemical method. Results: In the dark-avoidance and step-down tests, the performance record of rats with FGR was worse than that of control rats, and the behavior-trained rats was better than the untrained rats, when the FGR model and training factors were analyzed singly. The model factor and training factor had significant interacfion(P < 0.05). The expressions of NR2B and GIuR1 subunits in hippocampal CA1 and CA3 areas of rats with FGR reduced. In contrast, the expressions of GIuR1 and NR2B subunits in CA1 area of behavior-trained rats increased, when the FGR model and training factors were analyzed singly. Conclusion: These findings suggested that the effect of behavior training on the expressions of NR2B and GiuR1 subunits in CA1 area should be the mechanistic basis for the training-induced improvement in learning-memory abilities.

  17. Molecular characterization of ZzR1 resistance gene from Zingiber zerumbet with potential for imparting Pythium aphanidermatum resistance in ginger.

    Nair, R Aswati; Thomas, George


    Soft rot disease caused by the oomycete Pythium aphanidermatum (Edson) Fitzp. is the most economically significant disease of ginger (Zingiber officinale Rosc.) in tropical countries. All available ginger cultivars are susceptible to this pathogen. However a wild ginger relative viz., Zingiber zerumbet L. Smith, was identified as a potential soft rot resistance donor. In the present study, a putative resistance (R) gene designated, ZzR1 was isolated and characterized from Z. zerumbet using sequence information from Zingiber RGCs identified in our earlier experiments. Analysis of the 2280 bp segment revealed a 2157 bp open reading frame (ORF) encoding a putative cytoplasmically localized protein. The deduced ZzR1 protein shared high homology with other known R-genes belonging to the CC-NBS-LRR (coiled coil-nucleotide binding site-leucine rich repeat) class and had a calculated molecular weight of 84.61kDa. Real-time PCR analysis of ZzR1 transcription in Z. zerumbet following pathogen infection demonstrated activation at 3 hpi thus suggesting an involvement of ZzR1 in Z. zerumbet defense mechanism. Although many R-genes have been characterized from different taxa, none of them will help in the development of resistant ginger cultivars owing to the phenomenon of "Restricted Taxonomic Functionality" (RTF). Thus ZzR1 gene characterized from the resistant wild Zingiber accession represents a valuable genomic resource for ginger improvement programs. This first report on R-gene isolation from the Zingiber secondary gene pool is pivotal in designing strategies for engineering resistance in ginger, which is otherwise not amenable to conventional improvement programs owing to sexual reproduction barriers.

  18. Central injection of GalR1 agonist M617 facilitates GLUT4 expression in cardiac muscle of type 2 diabetic rats.

    Fang, Penghua; Shi, Mingyi; Zhu, Yan; Zhang, Zhenwen; Bo, Ping


    Although galanin has been shown to increase GLUT4 expression in the cardiac muscle of rats, there is no literature available about the effect of GalR1 on GLUT4 expression in the cardiac muscle of type 2 diabetic rats. The aim of this study was to determine whether intracerebroventricular injection of GalR1 agonist M617 would elevate GLUT4 expression in the cardiac muscle of type 2 diabetic rats. The rats tested were divided into four groups: rats from healthy and type 2 diabetic drug groups were injected with 10nM/kg/d M617 in 5μl artificial cerebrospinal fluid for 21days, while control received 5μl vehicle injections. The blood samples were analyzed for glucose and insulin concentration. Cardiac muscle was collected and processed for determination of GLUT4 mRNA expression and GLUT4 protein levels. The present findings showed that fasting blood glucose levels in both M617 treatment groups were lower compared with each control. The insulin levels in both M617 treatment groups were decreased compared with each control. Moreover, the GLUT4 content in the cardiac muscle in both drug groups was higher compared with each control. M617 treatment increased GLUT4 mRNA expression and GLUT4 protein levels compared with each control group. These observations suggest that GalR1 agonist M617, acting through its central GalR1, can promote GLUT4 expression and enhance GLUT4 content in the cardiac muscle of type 2 diabetic rats. Central GalR1 may play a significant role in regulation of glucose metabolic homeostasis in the cardiac muscle of type 2 diabetic rats.

  19. Relationship between motor complications of Parkinson's disease and phosphorylated GluR1Ser845%帕金森病运动并发症与谷氨酸受体亚型GluR1Ser845磷酸化关系的实验研究

    巴茂文; 刘振国; 孔敏; 陈生弟; 陆国强


    目的 探讨帕金森病(PD)长期左旋多巴治疗的运动并发症与纹状体神经元谷氨酸受体1的845位丝氨酸(GluR1Ser845)磷酸化的关系.方法 通过6-羟基多巴立体定向注射至大鼠前脑内侧前脑束建立PD动物模型,然后左旋多巴甲酯腹腔注射治疗(25 mg·kg-1·d-1,每天2次)22 d,评估旋转时间、关期发生频率情况;采用免疫荧光与蛋白印迹法检测纹状体区谷氨酸受体1(GluR1)亚细胞分布及GluR1Ser845磷酸化的表达情况.结果 PD大鼠长期应用左旋多巴处理后呈现旋转时间逐渐缩短、关期频率递增的趋势,与人类症状波动和开关现象具有相似特征.PD大鼠损伤侧纹状体细胞膜上GluR1和GluR1Ser845磷酸化的数量分别减少至73.0%±4.8%和42.0%±5.6%;长期左旋多巴处理后使损伤侧纹状体细胞膜上GluR1和GluR1Ser845磷酸化的数量分别增加至104.0%±5.5%和112.0%±3.4%;然而损伤侧纹状体GluR1总蛋白数量未发生明显变化.这些改变特异性发生在小清蛋白阳性的中间神经元上.结论 长期左旋多巴治疗的运动并发症可能与小清蛋白阳性的神经元上GluR1的亚细胞分布及GluR1Ser845磷酸化的改变有关.

  20. Effect of CRH and CRH-R1 at labor onset%CRH及其受体-1在分娩发动中的作用

    王瑜; 罗穗豫; 李睿; 董黎


    目的:比较早产及足月分娩剖宫产产妇血清肾上腺皮质激素释放激素( CRH)水平及子宫下段中促肾上腺皮质激素释放激素受体-1(CRH-R1)的表达情况,分析其在分娩发动中的作用。方法按难免早产、足月分娩发动、足月分娩未发动分为3组,术前测定血清中CRH水平,于剖宫产中取子宫下段平滑肌,用免疫组化的方法检测CRH-R1在子宫下段的表达。结果3组间CRH水平差异无统计学意义(P>0.05),均为孕期较高水平。难免早产组、足月分娩发动组CRH-R1在子宫下段肌层中呈高表达状,其表达指数明显高于足月分娩未发动组(t值分别为5.000、6.295,均P<0.05)。难免早产组、足月分娩发动组CRH-R1在子宫下段肌层中的表达差异无统计学意义(P>0.05)。结论 CRH为参与人类分娩启动因子之一。母体血浆CRH水平有望作为早产预测和预后指标,其作用的发挥还有赖于子宫体中CRH-R1的表达。%Objective To compare the level of serum corticotrophin-releasing hormone (CRH) and expression of corticotropin-releasing hormone receptor 1 ( CRH-R1) in lower uterine segment of pregnant women undergoing preterm and term birth cesarean section , and to analyze their role in laboronset .Methods Patients were divided into three groups according to inevitable premature birth , term labor onset and term labor without onset .Serum CRH level was tested before operation .In cesarean section myometrial samples were collected from the lower uterine segment to detect CRH-R1 expression with immunohistochemical method .Results CRH levels were not significantly different among three groups (P>0.05), and they were at relatively high level during pregnancy .In the group of inevitable premature birth and the group of term labor onset CRH-R1 was highly expressed in the lower uterine segment muscle layer , and its expression index was obviously higher than that of

  1. A Novel Role for the Receptor of the Complement Cleavage Fragment C5a, C5aR1, in CCR5-Mediated Entry of HIV into Macrophages.

    Moreno-Fernandez, Maria E; Aliberti, Julio; Groeneweg, Sander; Köhl, Jörg; Chougnet, Claire A


    The complement system is an ancient pattern recognition system that becomes activated during all stages of HIV infection. Previous studies have shown that C5a can enhance the infection of monocyte-derived macrophages and T cells indirectly through the production of interleukin (IL)-6 and tumor necrosis factor (TNF)-α and the attraction of dendritic cells. C5a exerts its multiple biologic functions mainly through activation of C5a receptor 1 (C5aR1). Here, we assessed the role of C5aR1 as an enhancer of CCR5-mediated HIV infection. We determined CCR5 and C5aR1 heterodimer formation in myeloid cells and the impact of C5aR1 blockade on HIV entry and genomic integration. C5aR1/CCR5 heterodimer formation was identified by immunoprecipitation and western blotting. THP-1 cells and monocyte-derived macrophages (MDM) were infected by R5 laboratory strains or HIV pseudotyped for the vesicular stomatitis virus (VSV) envelope. Levels of integrated HIV were measured by quantitative PCR after targeting of C5aR1 by a C5aR antagonist, neutralizing C5aR1 monoclonal antibody (mAb) or hC5a. C5aR1 was also silenced by specific siRNA prior to viral entry. We found that C5aR1 forms heterodimers with the HIV coreceptor CCR5 in myeloid cells. Targeting C5aR1 significantly decreased integration by R5 viruses but not by VSV-pseudotyped viruses, suggesting that C5aR1 is critical for viral entry. The level of inhibition achieved with C5aR1-blocking reagents was comparable to that of CCR5 antagonists. Mechanistically, C5aR1 targeting decreased CCR5 expression. MDM from CCR5Δ32 homozygous subjects expressed levels of C5aR1 similar to CCR5 WT individuals, suggesting that mere C5aR1 expression is not sufficient for HIV infection. HIV appeared to preferentially enter THP-1 cells expressing high levels of both C5aR1 and CCR5. Targeted reduction of C5aR1 expression in such cells reduced HIV infection by ~50%. Our data thus suggest that C5aR1 acts as an enhancer of CCR5-mediated HIV entry into

  2. A metallo-keratinase from a newly isolated Acinetobacter sp. R-1 with low collagenase activity and its biotechnological application potential in leather industry.

    Zhang, Rong-Xian; Gong, Jin-Song; Zhang, Dan-Dan; Su, Chang; Hou, Ying-Shuo; Li, Heng; Shi, Jin-Song; Xu, Zheng-Hong


    Microbial keratinase is a well-recognized enzyme that can specifically degrade insoluble keratins. A keratinase-producing bacterium was isolated from a duck ranch soil and identified as Acinetobacter sp. R-1 based on the biochemical characteristics and 16S rDNA gene sequencing. It showed high keratinase activity and low collagenase activity. The keratinase was purified to electrophoretic homogeneity with 6.69% recovery, 2.68-fold purification and an estimated molecular weight of 25 kDa. Additionally, the keratinase showed optimal activity at 50 °C and pH11. Keratinase activity of Acinetobacter sp. significantly increased in the presence of Li(+), Na(+), and Ca(2+), while it was completely inhibited by EDTA, indicating it was a metallo-keratinase. Moreover, the crude keratinase from Acinetobacter sp. R-1 could thoroughly depilate goat skin and simultaneously modify the wool surface, which indicated its applicable potential in leather and textile industries.

  3. Synthesis and pharmacological evaluation of 1-alkyl-N-[(1R)-1-(4-fluorophenyl)-2-methylpropyl]piperidine-4-carboxamide derivatives as novel antihypertensive agents.

    Watanuki, Susumu; Matsuura, Keisuke; Tomura, Yuichi; Okada, Minoru; Okazaki, Toshio; Ohta, Mitsuaki; Tsukamoto, Shin-ichi


    We synthesized and evaluated inhibitory activity against T-type Ca(2+) channels for a series of 1-alkyl-N-[(1R)-1-(4-fluorophenyl)-2-methylpropyl]piperidine-4-carboxamide derivatives. Structure-activity relationship studies have revealed that the isopropyl substituent at the benzylic position plays an important role in exerting potent inhibitory activity, and the absolute configuration of the benzylic position was found to be opposite that of mibefradil, which was first launched as a new class of T-type Ca(2+) channel blocker. Oral administration of N-[(1R)-1-(4-fluorophenyl)-2-methylpropyl]-1-[2-(3-methoxyphenyl)ethyl]piperidine-4-carboxamide (17f) lowered blood pressure in spontaneously hypertensive rats without inducing reflex tachycardia, an adverse effect often caused by traditional L-type Ca(2+) channel blockers.

  4. On Schr\\"odinger equation with potential U = - {\\alpha}r^{-1} + {\\beta}r + kr^{2} and the bi-confluent Heun functions theory

    Ovsiyuk, E; Veko, O


    It is shown that Schr\\"odinger equation with combination of three potentials U = - {\\alpha} r^{-1} + {\\beta} r + kr^{2}, Coulomb, linear and harmonic, the potential often used to describe quarkonium, is reduced to a bi-confluent Heun differential equation. The method to construct its solutions in the form of polynomials is developed, however with additional constraints in four parameters of the model, {\\alpha}, {\\beta}, k, l. The energy spectrum looks as a modified combination of oscillator and Coulomb parts.

  5. AMIGO3 is an NgR1/p75 co-receptor signalling axon growth inhibition in the acute phase of adult central nervous system injury.

    Zubair Ahmed

    Full Text Available Axon regeneration in the injured adult CNS is reportedly inhibited by myelin-derived inhibitory molecules, after binding to a receptor complex comprised of the Nogo-66 receptor (NgR1 and two transmembrane co-receptors p75/TROY and LINGO-1. However, the post-injury expression pattern for LINGO-1 is inconsistent with its proposed function. We demonstrated that AMIGO3 levels were significantly higher acutely than those of LINGO-1 in dorsal column lesions and reduced in models of dorsal root ganglion neuron (DRGN axon regeneration. Similarly, AMIGO3 levels were raised in the retina immediately after optic nerve crush, whilst levels were suppressed in regenerating optic nerves, induced by intravitreal peripheral nerve implantation. AMIGO3 interacted functionally with NgR1-p75/TROY in non-neuronal cells and in brain lysates, mediating RhoA activation in response to CNS myelin. Knockdown of AMIGO3 in myelin-inhibited adult primary DRG and retinal cultures promoted disinhibited neurite growth when cells were stimulated with appropriate neurotrophic factors. These findings demonstrate that AMIGO3 substitutes for LINGO-1 in the NgR1-p75/TROY inhibitory signalling complex and suggests that the NgR1-p75/TROY-AMIGO3 receptor complex mediates myelin-induced inhibition of axon growth acutely in the CNS. Thus, antagonizing AMIGO3 rather than LINGO-1 immediately after CNS injury is likely to be a more effective therapeutic strategy for promoting CNS axon regeneration when combined with neurotrophic factor administration.

  6. Cartilage quality in rheumatoid arthritis: comparison of T2* mapping, native T1 mapping, dGEMRIC, {delta}R1 and value of pre-contrast imaging

    Buchbender, Christian; Scherer, Axel; Kroepil, Patric; Quentin, Michael; Reichelt, Dorothea C.; Lanzman, Rotem S.; Mathys, Christian; Blondin, Dirk; Wittsack, Hans-Joerg; Antoch, Gerald; Miese, Falk [University Duesseldorf, Department of Diagnostic and Interventional Radiology, Medical Faculty, Duesseldorf (Germany); Koerbl, Birthe [Heinrich-Heine-University, Department of Endocrinology, Diabetology and Rheumatology, Medical Faculty, Duesseldorf (Germany); Heinrich-Heine-University, Leibniz Centre for Diabetes Research, Institute of Biometrics and Epidemiology, German Diabetes Centre, Duesseldorf (Germany); Bittersohl, Bernd; Zilkens, Christoph [Heinrich-Heine-University, Department of Orthopaedics, Medical Faculty, Duesseldorf (Germany); Hofer, Matthias [Heinrich-Heine-University, Medical Education Group, Medical School, Duesseldorf (Germany); Schneider, Matthias; Ostendorf, Benedikt [Heinrich-Heine-University, Department of Endocrinology, Diabetology and Rheumatology, Medical Faculty, Duesseldorf (Germany)


    To prospectively evaluate four non-invasive markers of cartilage quality - T2* mapping, native T1 mapping, dGEMRIC and {delta}R1 - in healthy volunteers and rheumatoid arthritis (RA) patients. Cartilage of metacarpophalangeal (MCP) joints II were imaged in 28 consecutive subjects: 12 healthy volunteers [9 women, mean (SD) age 52.67 (9.75) years, range 30-66] and 16 RA patients with MCP II involvement [12 women, mean (SD) age 58.06 (12.88) years, range 35-76]. Sagittal T2* mapping was performed with a multi-echo gradient-echo on a 3 T MRI scanner. For T1 mapping the dual flip angle method was applied prior to native T1 mapping and 40 min after gadolinium application (delayed gadolinium-enhanced MRI of cartilage, dGEMRIC, T1{sub Gd}). The difference in the longitudinal relaxation rate induced by gadolinium ({delta}R1) was calculated. The area under the receiver operating characteristic curve (AROC) was used to test for differentiation of RA patients from healthy volunteers. dGEMRIC (AUC 0.81) and {delta}R1 (AUC 0.75) significantly differentiated RA patients from controls. T2* mapping (AUC 0.66) and native T1 mapping (AUC 0.66) were not significantly different in RA patients compared to controls. The data support the use of dGEMRIC for the assessment of MCP joint cartilage quality in RA. T2* and native T1 mapping are of low diagnostic value. Pre-contrast T1 mapping for the calculation of {delta}R1 does not increase the diagnostic value of dGEMRIC. (orig.)

  7. Interaction between PLA2R1 and HLA-DQA1 variants associates with anti-PLA2R antibodies and membranous nephropathy.

    Lv, Jicheng; Hou, Wanyin; Zhou, Xujie; Liu, Gang; Zhou, Fude; Zhao, Na; Hou, Ping; Zhao, Minghui; Zhang, Hong


    Risk alleles at genome loci containing phospholipase A2 receptor 1 (PLA2R1) and HLA-DQA1 closely associate with idiopathic membranous nephropathy (IMN) in the European population, but it is unknown whether a similar association exists in the Chinese population and whether high-risk alleles promote the development of anti-PLA2R antibodies. Here, we genotyped 2132 Chinese individuals, including 1112 patients with IMN and 1020 healthy controls, for three single nucleotide polymorphisms (SNPs) within PLA2R1 and three SNPs within HLA genes. We also selected 71 patients, with varying genotypes, to assess for circulating anti-PLA2R antibody and for PLA2R expression in glomeruli. Three SNPs within PLA2R1 and one SNP within HLA-DQA1 strongly associated with IMN, and we noted gene-gene interactions involving these SNPs. Furthermore, these risk alleles strongly associated with the presence of anti-PLA2R antibodies and glomerular PLA2R expression. Among individuals who carried risk alleles for both genes, 73% had anti-PLA2R antibodies and 75% expressed PLA2R in glomeruli. In contrast, among individuals who carried protective genotypes of both genes, none had anti-PLA2R antibodies and glomerular expression of PLA2R was weak or absent. In conclusion, the interaction between PLA2R1 and HLA-DQA1 risk alleles associates with the development of IMN in the Chinese population. Individuals carrying risk alleles are predisposed to the generation of circulating anti-PLA2R autoantibodies, which may contribute to the development of IMN.

  8. Analysis of the APX, PGD1 and R1G1B constitutive gene promoters in various organs over three homozygous generations of transgenic rice plants.

    Park, Su-Hyun; Bang, Seung Woon; Jeong, Jin Seo; Jung, Harin; Redillas, Mark Christian Felipe Reveche; Kim, Hyung Il; Lee, Kang Hyun; Kim, Youn Shic; Kim, Ju-Kon


    We have previously characterized the constitutively active promoters of the APX, PGD1 and R1G1B genes in rice (Park et al. 2010 in J Exp Bot 61:2459-2467). To have potential crop biotechnology applications, gene promoters must be stably active over many generations. In our current study, we report our further detailed analysis of the APX, PGD1 and R1G1B gene promoters in various organs and tissues of transgenic rice plants for three (T₃₋₅) homozygous generations. The copy numbers in 37 transgenic lines that harbor promoter:gfp constructs were determined and promoter activities were measured by real-time qPCR. Analysis of the 37 lines revealed that 15 contained a single copy of one of the three promoter:gfp chimeric constructs. The promoter activity levels were generally higher in multi-copy lines, whereas variations in these levels over the T₃₋₅ generations studied were observed to be smaller in single-copy than in multi-copy lines. The three promoters were further found to be highly active in the whole plant body at both the vegetative and reproductive stages of plant growth, with the exception of the APX in the ovary and R1G1B in the pistil and filaments where zero or very low levels of activity were detected. Of note, the spatial activities of the PGD1 promoter were found to be strikingly similar to those of the ZmUbi1, a widely used constitutive promoter. Our comparison of promoter activities between T₃, T₄ and T₅ plants revealed that the APX, PGD1 and R1G1B promoters maintained their activities at comparable levels in leaves and roots over three homozygous generations and are therefore potentially viable alternative promoters for crop biotechnology applications.

  9. 两类非连通图(P2∨(Kn))(0,0,r1,0,…,0,rn)∪St( m)及(P2∨(Kn))(r1+a,r2,0,…,0)∪Gr的优美性%The Gracefulness of Two Kinds of Unconnected Graphs (P2∨(Kn))(0,0,r1,0,…,0,rn)∪St(m) and (P2∨(Kn))(r1 + a,r2,0,…,0)∪Gr

    吴跃生; 徐保根


    For natural numbers n, m and i∈ N, let Ki be an i-vertex complete graph, let Ka be the complement graph of graph Kn, A (m + 1) -vertex star tree is represented by St(m). Let Gr be a graceful graph with r-edges, let Pn be a n-vertex path and let P2 V Kn be the join graph of P2 and Kn. Two kinds of unconnected graphs (P2 ∨(Kn))(r1,r2,0,…,0)∪St(m) and (P2∨(Kn))(r1 +a,r2,0,…,0)∪Gr, are presented. It proves that the above two kinds of graphs are graceful graphs when n ≥ 2.%对自然数n,m,i∈N,设Ki表示i个顶点的完全图,(Kn)表示Kn的补图,St(m)表示m+1个顶点的星形树,G,为有r条边的优美图,Pn为n个节点的路,P2 ∨(Kn)是P2与(Kn)联图.给出了非连通图(P2 ∨(Kn))(r1,r2,0,…,0)∪St(m)及(P2∨(Kn))(r1 +a,r2,0,…,0)∪Gr的定义,并论证了当n≥2时,这两类图都是优美图.

  10. IL-10R1 S138G loss-of-function polymorphism is associated with extrapulmonary tuberculosis risk development in Tunisia.

    Ben-Selma, Walid; Ben-Abderrahmen, Yosra; Boukadida, Jalel; Harizi, Hedi


    There is considerable evidence that host genetic factors are important in determining susceptibility to mycobacterial infections. More recently, functional genetic mutations affecting IL-10 receptor 1 (IL-10R1) were described. In this study, we investigated the relationship of IL-10R1 S138G loss-of-function polymorphism (A536G: rs3135932) with susceptibility to active tuberculosis (TB) in Tunisian patients. A total of 168 patients with pulmonary TB, 55 with extrapulmonary TB, and 150 control subjects were studied. Genomic DNA samples were extracted from leukocytes and used to investigate S138G polymorphism in IL-10R1 gene by multiplex allele-specific polymerase chain reaction. Associations between G allele [odds ratio OR=5.01; 95% confidence intervals CI=2.58-9.77; P=10(-7)], GG genotypes [OR=9.06; 95% CI (1.58-67.33); correcting P-values using the Bonferroni method for multiple tests Pc=0.015] and AG genotype [OR=3.75; 95% CI (1.62-8.7); Pc=0.0012] with the risk development of active extrapulmonary TB were found. In contrast, the AA genotype was found to be associated with resistance to extrapulmonary TB [OR=0.19; 95% CI (0.09-0.42); Pc=6.10(-6)]. No association was found between S138G SNP and pulmonary TB. In conclusion, our study suggested the possible role of IL-10R1 S138G loss-of-function polymorphism in extrapulmonary TB susceptibility-resistance in Tunisia.

  11. TRH/TRH-R1 receptor signaling in the brain medulla as a pathway of vagally mediated gut responses during the cephalic phase.

    Taché, Yvette; Adelson, David; Yang, Hong


    Pavlov's seminal findings in the early twentieth century showed that the sight, smell or taste of food in dogs with chronic esophagostomy induces a vagal-dependent gastric acid secretion. These observations established the concept of the cephalic phase of digestion. Compelling experimental evidence in rats indicates that the three amino acid peptide thyrotropin-releasing hormone (TRH) expressed in the brainstem plays a key role in the vagal stimulation of gastric function. Neurons in the dorsal motor nucleus of the vagus (DMN) expressed TRH receptor subtype (TRH-R1) and received efferent input from TRH containing fibers arising from TRH synthesizing neurons in the raphe pallidus, raphe obscurus, and the parapyramidal regions. TRH microinjected into the DMN or intracisternally excites the firing of DMN neurons and stimulates efferent activity in the gastric branch of the vagus nerve and gastric myenteric cholinergic neurons. At the functional level, this results in a vagally-mediated and atropine-sensitive stimulation of gastric epithelial and endocrine cells secreting acid, pepsin, serotonin, histamine and ghrelin, and enteric neurons leading to increased gastric motility and emptying. Importantly, the blockade of TRH or TRH-R1 in the brainstem by pretreatment into the cisterna magna or the DMN with TRH antibody or TRH-R1 oligodeoxynucleotide antisense respectively abolishes the stimulation of gastric acid induced by sham-feeding. The gastric response to TRH injected into the DMN is potentiated by serotonin and the proTRH flanking peptide, Ps4 and suppressed by a number of brainstem peptides and cytokines activated during stress or immune response and inhibiting food intake and gastric acid secretion. These convergent data strongly support a physiological involvement of TRH signaling pathway in the brainstem to stimulate vagal activity and identified TRH-TRH-R1 system as a major effector in the dorsal vagal complex to drive the vagally mediated gut response

  12. Enhanced plasma ghrelin levels in Helicobacter pylori-colonized,interleukin-1-receptor type 1-homozygous knockout (IL-1R1-/-) mice

    Yuka Abiko; Hidekazu Suzuki; Tatsuhiro Masaoka; Sachiko Nomura; Kumiko Kurabayashi; Hiroshi Hosoda; Kenji Kangawa; Toshifumi Hibi


    AIM: Ghrelin is an endogenous ligand for the growth hormone secretagogue receptor, and it plays a role in stimulating the growth hormone secretion, food intake,body weight gain and gastric motility. Eradication of Helicobacter pylori(H pylori) was shown to be associated with increase of the body weight. On the other hand, H pylori infection evokes the release of gastric IL-1β. The present study was designed to investigate the involvement of the gastric IL-1 signal in the ghrelin dynamics in H pyloricolonized mice.METHODS: Twelve-week-old female IL-1-receptor type 1-homozygous-knockout mice (IL-1R1-/-) and their wild-type littermates (WT) were orally inoculated with H pylori (Hp group), while other cohorts received oral inoculation of culture medium (Cont group). Thirteen weeks after the inoculation, the mice were examined. The plasma and stomach ghrelin levels and the gastric preproghrelin mRNA were measured.RESULTS: Although the WT mice with H pylori infection showed a significantly decreased body weight as compared with that of the animals without H pylori infection,H pylori infection did not influence the body weight of the IL-1R1-knockout (IL-1R1-/-) mice. In the H pylori-infected IL-1R1-/-mice, the total and active ghrelin levels in the plasma were significantly increased, and the gastric ghrelin level was decreased. No significant differences were noted in the gastric preproghrelin mRNA expression.CONCLUSION: Ghrelin secretion triggered by H pylori infection might be suppressed by IL-1β, the release of which is also induced by the infection, resulting in the body weight loss of mice with H pylori infection.

  13. Extinction of morphine-dependent conditioned behavior is associated with increased phosphorylation of the GluR1 subunit of AMPA receptors at hippocampal synapses

    Billa, Sophie K.; Sinha, Namita; Rudrabhatla, Sri Rajyalakshmi; Morón, Jose A.


    In abstinent opiate addicts, relapse can be triggered by exposure to environmental cues associated with drug use; thus, the disruption of these learned associations may be an effective approach for reducing relapse. Interestingly, glutamatergic systems are thought to be involved in opiate-induced behavioral plasticity. In this study, changes in expression and phosphorylation levels of AMPA glutamate receptor subunits (GluR1, GluR2) in the hippocampus were investigated in rats showing a condit...

  14. Essential Genes for In Vitro Growth of the Endophyte Herbaspirillum seropedicae SmR1 as Revealed by Transposon Insertion Site Sequencing

    Rosconi, Federico; de Vries, Stefan P. W.; Baig, Abiyad; Fabiano, Elena; Grant, Andrew J


    ABSTRACT The interior of plants contains microorganisms (referred to as endophytes) that are distinct from those present at the root surface or in the surrounding soil. Herbaspirillum seropedicae strain SmR1, belonging to the betaproteobacteria, is an endophyte that colonizes crops, including rice, maize, sugarcane, and sorghum. Different approaches have revealed genes and pathways regulated during the interactions of H. seropedicae with its plant hosts. However, functional genomic analysis o...

  15. Expression of conserved signalling pathway genes during spontaneous vascular differentiation of R1 embryonic stem cells and in Py-4-1 endothelial cells

    Kavitha Siva; K Gokul; Maneesha S Inamdar


    Embryonic stem (ES) cells are an invaluable model for identifying subtle phenotypes as well as severe outcomes of perturbing gene function that may otherwise result in lethality. However, though ES cells of different origins are regarded as equally pluripotent, their in vitro differentiation potential varies, suggesting that their response to developmental signals is different. The R1 cell line is widely used for gene manipulation due to its good growth characteristics and highly efficient germline transmission. Hence, we analysed the expression of Notch, Wnt and Sonic Hedgehog (Shh) pathway genes during differentiation of R1 cells into early vascular lineages. Notch-, Wnt-and Shh-mediated signalling is important during embryonic development. Regulation of gene expression through these signalling molecules is a frequently used theme, resulting in context-dependent outcomes during development. Perturbing these pathways can result in severe and possibly lethal developmental phenotypes often due to primary cardiovascular defects. We report that during early spontaneous differentiation of R1 cells, Notch-1 and the Wnt target Brachyury are active whereas the Shh receptor is not detected. This expression pattern is similar to that seen in a mouse endothelial cell line. This temporal study of expression of genes representative of all three pathways in ES cell differentiation will aid in further analysis of cell signalling during vascular development.

  16. Low enriched uranium UAl{sub X}-Al targets for the production of Molybdenum-99 in the IEA-R1 and RMB reactors

    Domingos, Douglas B.; Silva, Antonio T. e; Joao, Thiago G.; Silva, Jose Eduardo R. da, E-mail: teixeira@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Nishiyama, Pedro J.B. de O., E-mail: [Centro Tecnologico da Marinha em Sao Paulo (CTMSP), SP (Brazil)


    The IEA-R1 reactor of IPEN/CNEN-SP in Brazil is a pool type research reactor cooled and moderated by demineralized water and having Beryllium and Graphite as reflectors. In 1997 the reactor received the operating licensing for 5 MW. A new research reactor is being planned in Brazil to replace the IEA-R1 reactor. This new reactor, the Brazilian Multipurpose Reactor (RMB), planned for 30 MW, is now in the conception design phase. Low enriched uranium (LEU) (<20% {sup 235}U) UAl{sub x} dispersed in Al targets are being considered for production of Molybdenum-99 ({sup 99}Mo) by fission. Neutronic and thermal-hydraulics calculations were performed, respectively, to compare the production of {sup 99}Mo for these targets in IEA-R1 reactor and RMB and to determine the temperatures achieved in the UAl{sub x}-Al targets during irradiation. For the neutronic calculations were utilized the computer codes HAMMER-TECHNION, CITATION and SCALE and for the thermal-hydraulics calculations was utilized the computer code MTRCR-IEAR1. (author)

  17. Relaxor ferroeletric behavior in S r1 -xP rxTi O3 : Cooperation between polar and antiferrodistortive instabilities

    Checchia, Stefano; Allieta, Mattia; Coduri, Mauro; Brunelli, Michela; Scavini, Marco


    Chemical doping at the Sr and Ti sites is a feasible way to alter the quantum paraelectric state of SrTi O3 perovskite. Doping with Pr is known to induce relaxor ferroelectricity at room temperature in the S r1 -xP rxTi O3 solid solution. The relationship between its dielectric properties and structural phase transition has been debated, but no definitive structural argument has been proposed. Here we present a systematic structural study of S r1 -xP rxTi O3 (0.020 ≤x ≤0.150 ). We establish the structural phase diagram using high-resolution x-ray powder diffraction by finding the antiferrodistortive structural phase transitions for all the compositions studied. By using pair distribution function analysis, we show the mismatch between local and long-range structures in terms of increased local order parameters. Finally, we propose a correlation between the local structural order parameters and the emergence of hard polar modes as found by Raman spectroscopy. Our results are quantitatively consistent with recent theoretical calculations showing that the increase of local tetragonality and local octahedral tilting above a critical value in fact underlie the polar instability. This confirms that structural orders involving both polar and antiferrodistortive characters compete and cooperate at different levels, promoting ferroelectricity in S r1 -xP rxTi O3 .

  18. Changes in dopamine D2 and GluR-1 glutamate receptor mRNAs in the rat brain after treatment with phencyclidine.



    Full Text Available In situ hybridization of slide-mounted brain sections from rats subjected to acute and chronic phencyclidine treatment was carried out using synthetic oligonucleotides complementary to dopamine D2-receptor and non-N-methyl-D-aspartate (NMDA glutamate-receptor-subunit (GluR-1 mRNAs. There was no significant difference in either the D2-receptor or the GluR-1 mRNA levels in any brain region of the acute phencyclidine (10 mg/kg-treated and control groups. However, chronic administration of phencyclidine (10 mg/kg/day, 14 days significantly decreased the dopamine D2-receptor mRNA level in the caudate-putamen (by 27%, P < 0.01 and significantly increased the GluR-1 mRNA level in the prefrontal cortex (by 29%, P < 0.001. These results suggest that the chronic pharmaco-behavioral effects of phencyclidine may involve expression of both dopamine- and non-NMDA glutamate-receptor mRNAs.

  19. Microbial production of the aromatic building-blocks (S)-styrene oxide and (R)-1,2-phenylethanediol from renewable resources.

    McKenna, Rebekah; Pugh, Shawn; Thompson, Brian; Nielsen, David R


    (S)-Styrene oxide and (R)-1,2-phenylethanediol are chiral aromatic molecular building blocks used commonly as precursors to pharmaceuticals and other specialty chemicals. Two pathways have been engineered in Escherichia coli for their individual biosynthesis directly from glucose. The novel pathways each constitute extensions of the previously engineered styrene pathway, developed by co-expressing either styrene monooxygenase (SMO) or styrene dioxygenase (SDO) to convert styrene to (S)-styrene oxide and (R)-1,2-phenylethanediol, respectively. StyAB from Pseudomonas putida S12 was determined to be the most effective SMO. SDO activity was achieved using NahAaAbAcAd of Pseudomonas sp. NCIB 9816-4, a naphthalene dioxygenase with known broad substrate specificity. Production of phenylalanine, the precursor to both pathways, was systematically enhanced through a number of mutations, most notably via deletion of tyrA and over-expression of tktA. As a result, (R)-1,2-phenylethanediol reached titers as high as 1.23 g/L, and at 1.32 g/L (S)-styrene oxide titers already approach their toxicity limit. As with other aromatics, product toxicity was strongly correlated with a model of membrane accumulation and disruption. This study additionally demonstrates that greater flux through the styrene pathway can be achieved if its toxicity is addressed, as achieved in this case by reacting styrene to less toxic products. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. RNA-Sequencing Reveals the Progression of Phage-Host Interactions between φR1-37 and Yersinia enterocolitica.

    Leskinen, Katarzyna; Blasdel, Bob G; Lavigne, Rob; Skurnik, Mikael


    Despite the expanding interest in bacterial viruses (bacteriophages), insights into the intracellular development of bacteriophage and its impact on bacterial physiology are still scarce. Here we investigate during lytic infection the whole-genome transcription of the giant phage vB_YecM_φR1-37 (φR1-37) and its host, the gastroenteritis causing bacterium Yersinia enterocolitica. RNA sequencing reveals that the gene expression of φR1-37 does not follow a pattern typical observed in other lytic bacteriophages, as only selected genes could be classified as typically early, middle or late genes. The majority of the genes appear to be expressed constitutively throughout infection. Additionally, our study demonstrates that transcription occurs mainly from the positive strand, while the negative strand encodes only genes with low to medium expression levels. Interestingly, we also detected the presence of antisense RNA species, as well as one non-coding intragenic RNA species. Gene expression in the phage-infected cell is characterized by the broad replacement of host transcripts with phage transcripts. However, the host response in the late phase of infection was also characterized by up-regulation of several specific bacterial gene products known to be involved in stress response and membrane stability, including the Cpx pathway regulators, ATP-binding cassette (ABC) transporters, phage- and cold-shock proteins.

  1. RNA-Sequencing Reveals the Progression of Phage-Host Interactions between φR1-37 and Yersinia enterocolitica

    Katarzyna Leskinen


    Full Text Available Despite the expanding interest in bacterial viruses (bacteriophages, insights into the intracellular development of bacteriophage and its impact on bacterial physiology are still scarce. Here we investigate during lytic infection the whole-genome transcription of the giant phage vB_YecM_φR1-37 (φR1-37 and its host, the gastroenteritis causing bacterium Yersinia enterocolitica. RNA sequencing reveals that the gene expression of φR1-37 does not follow a pattern typical observed in other lytic bacteriophages, as only selected genes could be classified as typically early, middle or late genes. The majority of the genes appear to be expressed constitutively throughout infection. Additionally, our study demonstrates that transcription occurs mainly from the positive strand, while the negative strand encodes only genes with low to medium expression levels. Interestingly, we also detected the presence of antisense RNA species, as well as one non-coding intragenic RNA species. Gene expression in the phage-infected cell is characterized by the broad replacement of host transcripts with phage transcripts. However, the host response in the late phase of infection was also characterized by up-regulation of several specific bacterial gene products known to be involved in stress response and membrane stability, including the Cpx pathway regulators, ATP-binding cassette (ABC transporters, phage- and cold-shock proteins.

  2. The bidirectionality of motor learning in the vestibulo-ocular reflex is a function of cerebellar mGluR1 receptors.

    Titley, Heather K; Heskin-Sweezie, Raquel; Broussard, Dianne M


    Bidirectional changes in synaptic transmission have the potential to optimize the control of movement. However, it can be difficult to establish a causal relationship between the bidirectionality of synaptic plasticity and bidirectional changes in the speed of actual movements. We asked whether metabotropic glutamate receptor 1 (mGluR1) receptors, which participate in cerebellar long-term depression (LTD), are necessary for bidirectional motor learning in the vestibulo-ocular reflex (VOR). Cerebellar LTD and long-term potentiation (LTP) are thought to cause increases and decreases, respectively, in the gain of the VOR; the direction of learning depends on the behavioral protocol. We injected either the mGluR1 agonist (S)-DHPG or the antagonist YM 298198 bilaterally into the flocculus of alert cats, and then induced motor learning. In the presence of YM 298198, the VOR gain decreased in gain-up, as well as in gain-down protocols. (S)-DHPG augmented gain-up learning. Gain-down learning was not significantly affected by either drug. These results supported the hypothesis that gain-up learning relies on cerebellar LTD, but gain-down learning relies on a different mechanism. In the absence of mGluR1 activity, cerebellar LTD may be replaced with LTP, permitting learning in only one direction.

  3. Lack of association between polymorphisms of the IL18R1 and IL18RAP genes and cardiovascular risk: the MORGAM Project

    Nicaud Viviane


    Full Text Available Abstract Background Interleukin-18 is a pro-inflammatory cytokine suspected to be associated with atherosclerosis and its complications. We had previously shown that one single nucleotide polymorphism (SNP of the IL18 gene was associated with cardiovascular disease (CVD through an interaction with smoking. As a further step for elucidating the contribution of the IL-18 pathway to the etiology of CVD, we here investigated the association between the genetic variability of two IL-18 receptor genes, IL18R1 and IL18RAP, with the risk of developing CVD. Methods Eleven tagging SNPs, 5 in IL18R1 and 6 in IL18RAP, characterizing the haplotypic variability of the corresponding genes; were genotyped in 5 European prospective CVD cohorts including 1416 cases and 1772 non-cases, as part of the MORGAM project. Both single-locus and haplotypes analyses were carried out to investigate the association of these SNPs with CVD. Results We did not find any significant differences in allele, genotype and haplotype frequencies between cases and non-cases for either of the two genes. Moreover, the search for interactions between SNPs located in different genes, including 5 IL18 SNPs previously studied in the MORGAM project, and between SNPs and environmental factors remained unfruitful. Conclusion Our analysis suggests that the variability of IL18R1 and IL18RAP genes are unlikely to contribute to modulate the risk of CVD.

  4. Immunohistochemical localization of galanin receptors (GAL-R1, GAL-R2, and GAL-R3) on myenteric neurons from the sheep and dog stomach.

    Arciszewski, Marcin Bartłomiej; Barabasz, Sylwester; Całka, Jarosław


    Galanin exerts its biological activities (inhibitory or excitatory) via three different G protein-coupled receptors. In the present study, double immunocytochemical labeling was used to localize GAL-R1, GAL-R2 and GAL-R3 on PGP 9.5-positive myenteric neurons from the dog and sheep stomach/forestomachs. In both species, the occurrence of galanin in neurons and nerve fibers of gastric ganglia was also studied. Myenteric ganglia of the dog stomach were supplied with numerous, mainly varicose, galanin-immunoreactive (IR) nerve terminals whereas the frequency of galanin-positive nerve fibers in myenteric ganglia of the ovine stomach and forestomachs was moderate. The number of PGP 9.5-IR/galanin-IR myenteric neurons was significantly lower in the dog stomach (12.3+/-1.3%) as compared to the sheep rumen (20.1+/-0.7%), omasum (19.5+/-2.9%), abomasum (23.8+/-1.2%) but not reticulum (8.1+/-0.8%). In the canine stomach the frequencies of GAL-R1, GAL-R2 and GAL-R3 expressing myenteric neurons were statistically equivalent (4.4+/-0.9%, 3.5+/-0.7% and 3.1+/-0.5%, respectively). Immunoreactivity to GAL-R1 was absent in myenteric ganglia from the ovine rumen, reticulum as well as omasum. GAL-R1 was localized on 0.5+/-0.3% of myenteric perikarya from the abomasum. GAL-R2 bearing myenteric neurons were localized in the ovine rumen (0.6+/-0.3%), reticulum (0.5+/-0.3%), omasum (1.0+/-0.2%) and abomasum (1.1+/-0.3%). The percentages of PGP 9.5-IR/GAL-R3-IR neurons were 0.8+/-0.2% in the rumen, 0.6+/-0.3% in the reticulum, 0.7+/-0.2% in the omasum and 0.9+/-0.3% in the abomasum. In all compartments of the sheep stomach, the proportions of GAL-R1, GAL-R2 and GAL-R3 expressing neurons were significantly lower when compared to analogous neuronal subpopulations present in the dog. It is suggested that, although endogenous galanin may potentially inhibit or stimulate the activity of sparse gastric enteric neurons, its general role in indirect mediation of gastric motility and/or secretion

  5. GR SUMOylation and formation of an SUMO-SMRT/NCoR1-HDAC3 repressing complex is mandatory for GC-induced IR nGRE-mediated transrepression

    Hua, Guoqiang; Paulen, Laetitia; Chambon, Pierre


    Unique among the nuclear receptor superfamily, the glucocorticoid (GC) receptor (GR) can exert three distinct transcriptional regulatory functions on binding of a single natural (cortisol in human and corticosterone in mice) and synthetic [e.g., dexamethasone (Dex)] hormone. The molecular mechanisms underlying GC-induced positive GC response element [(+)GRE]-mediated activation of transcription are partially understood. In contrast, these mechanisms remain elusive for GC-induced evolutionary conserved inverted repeated negative GC response element (IR nGRE)-mediated direct transrepression and for tethered indirect transrepression that is mediated by DNA-bound NF-κB/activator protein 1 (AP1)/STAT3 activators and instrumental in GC-induced anti-inflammatory activity. We demonstrate here that SUMOylation of lysine K293 (mouse K310) located within an evolutionary conserved sequence in the human GR N-terminal domain allows the formation of a GR-small ubiquitin-related modifiers (SUMOs)-NCoR1/SMRT-HDAC3 repressing complex mandatory for GC-induced IR nGRE-mediated direct repression in vitro, but does not affect transactivation. Importantly, these results were validated in vivo: in K310R mutant mice and in mice ablated selectively for nuclear receptor corepressor 1 (NCoR1)/silencing mediator for retinoid or thyroid-hormone receptors (SMRT) corepressors in skin keratinocytes, Dex-induced direct repression and the formation of repressing complexes on IR nGREs were impaired, whereas transactivation was unaffected. In mice selectively ablated for histone deacetylase 3 (HDAC3) in skin keratinocytes, GC-induced direct repression, but not bindings of GR and of corepressors NCoR1/SMRT, was abolished, indicating that HDAC3 is instrumental in IR nGRE-mediated repression. Moreover, we demonstrate that the binding of HDAC3 to IR nGREs in vivo is mediated through interaction with SMRT/NCoR1. We also show that the GR ligand binding domain (LBD) is not required for SMRT

  6. GR SUMOylation and formation of an SUMO-SMRT/NCoR1-HDAC3 repressing complex is mandatory for GC-induced IR nGRE-mediated transrepression.

    Hua, Guoqiang; Paulen, Laetitia; Chambon, Pierre


    Unique among the nuclear receptor superfamily, the glucocorticoid (GC) receptor (GR) can exert three distinct transcriptional regulatory functions on binding of a single natural (cortisol in human and corticosterone in mice) and synthetic [e.g., dexamethasone (Dex)] hormone. The molecular mechanisms underlying GC-induced positive GC response element [(+)GRE]-mediated activation of transcription are partially understood. In contrast, these mechanisms remain elusive for GC-induced evolutionary conserved inverted repeated negative GC response element (IR nGRE)-mediated direct transrepression and for tethered indirect transrepression that is mediated by DNA-bound NF-κB/activator protein 1 (AP1)/STAT3 activators and instrumental in GC-induced anti-inflammatory activity. We demonstrate here that SUMOylation of lysine K293 (mouse K310) located within an evolutionary conserved sequence in the human GR N-terminal domain allows the formation of a GR-small ubiquitin-related modifiers (SUMOs)-NCoR1/SMRT-HDAC3 repressing complex mandatory for GC-induced IR nGRE-mediated direct repression in vitro, but does not affect transactivation. Importantly, these results were validated in vivo: in K310R mutant mice and in mice ablated selectively for nuclear receptor corepressor 1 (NCoR1)/silencing mediator for retinoid or thyroid-hormone receptors (SMRT) corepressors in skin keratinocytes, Dex-induced direct repression and the formation of repressing complexes on IR nGREs were impaired, whereas transactivation was unaffected. In mice selectively ablated for histone deacetylase 3 (HDAC3) in skin keratinocytes, GC-induced direct repression, but not bindings of GR and of corepressors NCoR1/SMRT, was abolished, indicating that HDAC3 is instrumental in IR nGRE-mediated repression. Moreover, we demonstrate that the binding of HDAC3 to IR nGREs in vivo is mediated through interaction with SMRT/NCoR1. We also show that the GR ligand binding domain (LBD) is not required for SMRT

  7. Exendin-4 promotes the membrane trafficking of the AMPA receptor GluR1 subunit and ADAM10 in the mouse neocortex.

    Ohtake, Nobuaki; Saito, Mieko; Eto, Masaaki; Seki, Kenjiro


    Glucagon-like peptide-1 (GLP-1) is a novel treatment modality for type 2 diabetes mellitus. However, GLP-1 has been suggested as a therapeutic target for Alzheimer's disease (AD). In rodent studies, GLP-1 reduces amyloid beta (Aβ) and facilitates synaptic plasticity. Therefore, in the present study, we investigated how GLP-1 facilitates synaptic plasticity and reduces the Aβ in vivo. Exendin-4, a GLP-1 receptor agonist that can cross the blood brain barrier, was subcutaneously administered to adult mice. We then extracted the total and the plasma membrane proteins from the mouse neocortex. Exendin-4 significantly increased the phosphorylation level of cAMP response element-binding protein (CREB). Consistently, the expression level of brain-derived neurotrophic factor (BDNF), a transcriptional target of CREB, was increased. Furthermore, exendin-4 increased the membrane protein level of the AMPA receptor GluR1 subunit and postsynaptic density protein-95 (PSD-95), whereas GluR2 was unaffected. These exendin-4-dependent increases in membrane GluR1, total PSD-95 and BDNF were abrogated by pretreatment with temozolomide (TMZ), a DNA-alkylating agent, indicating that these alterations were dependent on exendin-4-induced transcriptional activity. In addition, we found that exendin-4 increased the level of the α-C terminal fragment (α-CTF) of amyloid precursor protein (APP). Furthermore, protein levels of both mature and immature ADAM10, the α-secretase of APP in the plasma membrane, were increased, whereas the total mature and immature ADAM10 levels were unchanged. These exendin-4-dependent increases in α-CTF and ADAM10 were not affected by TMZ. These findings suggested that GLP-1 facilitates the GluR1 membrane insertion through CREB activation and increases α-secretase activity through ADAM10 membrane trafficking. Upregulation of GluR1 and ADAM10 at the plasma membrane were also observed in mice with intracerebroventricular administration of Aβ oligomer

  8. Association of IL18R1 gene rs1O3513O PolymorPhism with schizoPhrenia%IL18R1基因多态性与中国汉族精神分裂症的关联性分析

    孙莉; 程灶火; 张付全; 徐勇


    目的:探讨 IL18受体 IL18R1基因多态性与中国汉族精神分裂症的关系。方法:采用病例对照研究方法,选取1174例精神分裂症患者(病例组)与1120名正常健康对照者(对照组),利用连接酶检测-聚合酶链反应(LDR-PCR)方法对 IL18R1基因多态位点 rs1035130进行基因分型,观察其等位基因及基因型与精神分裂症发病风险的关系。结果:rs1035130的基因型与等位基因频率在病例组与正常对照组中的分布差异无统计学意义(P ﹥0.05)。结论:IL18R1基因 rs1035130多态性可能与中国汉族精神分裂症无关。%ObJective:To explore the association between IL18R1( IL18receptor1)polymorphism and schizophrenia. Method:rs1035130 was genotyped by LDR-PCR methods,and the association of its polymor-phism and schizophrenia risk was analyzed in a case-control study with 1174 schizophrenic patients and 1120 healthy controls. Results:No significant differences for genotype and allete gene in IL18R1 gene were presen-ted between schizophrenic patients and healthy controls(P ﹥ 0. 05). Conclusion:IL18R1 gene rs1035130 may not relate to schizophrenia in Chinese Han population.

  9. Levodopa/benserazide microspheres reduced levodopa-induced dyskinesia by downregulating phosphorylated GluR1 expression in 6-OHDA-lesioned rats

    Song L,Yuan W


    . Abnormal involuntary movements were measured in rats on days 1, 5, 10, 15, and 20 during the treatment. The levels of GluR1 at serine-831 (pGluR1S831 and serine-845 (pGluR1S845 were determined by Western blot. Arc and proenkephalin (Penk levels were measured by real-time polymerase chain reaction.Results: Three-week levodopa plus benserazide treatment induced dyskinesia in PD rats. Levodopa/benserazide-loaded microsphere-treated dyskinetic rats showed lower AIM scores than levodopa plus benserazide-treated dyskinetic rats. Microsphere treatment downregulated the phosphrylated levels of pGluR1S831 and pGluR1S845 in the striatum of dyskinetic rats. In addition, microsphere treatment reduced the levels of Arc and Penk.Conclusion: These data indicated that levodopa/benserazide-loaded microspheres could be used to ameliorate the expression of LID by reducing the expression of pGluR1S831 and pGluR1S845 as well as Arc and Penk.Keywords: levodopa/beserazide-loaded microspheres, Arc, proenkephalin

  10. Arsenic sulfide promotes apoptosis in retinoid acid resistant human acute promyelocytic leukemic NB4-R1 cells through downregulation of SET protein.

    Yuwang Tian

    Full Text Available Tetra-arsenic tetra-sulfide (As4S4 is an arsenic compound with anti-tumor activity, especially in acute promyelocytic leukemia (APL that are resistant to retinoic acid (RA. Although recent studies revealed that the therapeutic action of As4S4 is closely associated with the induction of cellular apoptosis, the exact molecular mechanism of action of As4S4 in RA-resistant APL remains to be clarified. In this study, we found that As4S4-induced apoptosis was accompanied by reduced mRNA and protein expression of SET gene in RA-resistant NB4-R1 cells. Moreover, RNAi knockdown of SET gene further promoted As4S4-induced apoptosis, while SET over-expression inhibited it, suggesting that As4S4 induces apoptosis through the reduction of SET protein in NB4-R1 cells. We also demonstrated that the knockdown of SET gene resulted in the upregulation of protein phosphatase 2 (PP2A expression and the downregulation of promyelocytic leukemia and retinoic acid receptor α fusion gene (PML-RARα expression, which were enhanced by As4S4 treatments. By contrast, over-expression of SET gene resulted in PP2A downregulation and PML-RARα upregulation, which were abolished by As4S4 pretreatment. Since PP2A is a pro-apoptotic factor and PMLRARα is an anti-apoptotic factor, our results suggest that As4S4-induced apoptosis in NB4-R1 cells is through the downregulation of SET protein expression, which in turn increases PP2A and reduces PML-RARα expressions to lead to cell apoptosis.

  11. Consolidation of remote fear memories involves Corticotropin-Releasing Hormone (CRH) receptor type 1-mediated enhancement of AMPA receptor GluR1 signaling in the dentate gyrus.

    Thoeringer, Christoph K; Henes, Kathrin; Eder, Matthias; Dahlhoff, Maik; Wurst, Wolfgang; Holsboer, Florian; Deussing, Jan M; Moosmang, Sven; Wotjak, Carsten T


    Persistent dreadful memories and hyperarousal constitute prominent psychopathological features of posttraumatic stress disorder (PTSD). Here, we used a contextual fear conditioning paradigm to demonstrate that conditional genetic deletion of corticotropin-releasing hormone (CRH) receptor 1 within the limbic forebrain in mice significantly reduced remote, but not recent, associative and non-associative fear memories. Per os treatment with the selective CRHR1 antagonist DMP696 (3 mg/kg) attenuated consolidation of remote fear memories, without affecting their expression and retention. This could be achieved, if DMP696 was administered for 1 week starting as late as 24 h after foot shock. Furthermore, by combining electrophysiological recordings and western blot analyses, we demonstrate a delayed-onset and long-lasting increase in AMPA receptor (AMPAR) GluR1-mediated signaling in the dentate gyrus (DG) of the dorsal hippocampus 1 month after foot shock. These changes were absent from CRHR1-deficient mice and after DMP696 treatment. Inactivation of hippocampal GluR1-containing AMPARs by antisense oligonucleotides or philantotoxin 433 confirmed the behavioral relevance of AMPA-type glutamatergic neurotransmission in maintaining the high levels of remote fear in shocked mice with intact CRHR1 signaling. We conclude that limbic CRHR1 receptors enhance the consolidation of remote fear memories in the first week after foot shock by increasing the expression of Ca(2+)-permeable GluR1-containing AMPARs in the DG. These findings suggest both receptors as rational targets for the prevention and therapy, respectively, of psychopathology associated with exaggerated fear memories, such as PTSD.

  12. Association of cord blood des-acyl ghrelin with birth weight, and placental GHS-R1 receptor expression in SGA, AGA, and LGA newborns.

    González-Domínguez, Martha I; Lazo-de-la-Vega-Monroy, Maria-Luisa; Zaina, Silvio; Sabanero, Myrna; Daza-Benítez, Leonel; Malacara, Juan Manuel; Barbosa-Sabanero, Gloria


    Although ghrelin in cord blood has been associated to birth weight, its role in fetal and postnatal growth has not been elucidated. The aim of this study was to analyze total ghrelin, acyl ghrelin (AG), and des-acyl ghrelin (DAG) in cord blood of newborns with idiopathic birth weight alterations, and to evaluate protein expression of placental GHS-R1, in order to investigate their correlation with birth weight and placental weight. We performed a cross-sectional comparative study in umbilical cord blood and placentas from healthy mothers of SGA, AGA, and LGA (small, adequate and large for gestational age) term newborns (n = 20 per group). Cord blood total ghrelin, AG, and DAG were measured by ELISA, and placental GHS-R1 expression was evaluated by Western blot. Cord blood DAG was higher in SGA compared to AGA newborns (902.1 ± 109.1 and 597.4 ± 58.2 pg/ml, respectively, p = 0.01) while LGA and AGA showed similar values (627.2 ± 76.4 pg/ml for LGA, p = 0.80). DAG negatively correlated with birthweight (r = -0.31, p = 0.02) and placental weight (r = -0.33, p = 0.02). No differences in AG or total ghrelin were found. GHS-R1 protein in placenta was not differentially expressed among SGA, AGA, and LGA. Our results suggest a role of DAG in intrauterine growth. Further studies are needed in order to elucidate the mechanisms by which DAG participates in fetal growth.

  13. Ghrelin accelerates wound healing through GHS-R1a-mediated MAPK-NF-κB/GR signaling pathways in combined radiation and burn injury in rats.

    Liu, Cong; Huang, Jiawei; Li, Hong; Yang, Zhangyou; Zeng, Yiping; Liu, Jing; Hao, Yuhui; Li, Rong


    The therapeutic effect of ghrelin on wound healing was assessed using a rat model of combined radiation and burn injury (CRBI). Rat ghrelin, anti-rat tumor necrosis factor (TNF) α polyclonal antibody (PcAb), or selective antagonists of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK), and growth hormone secretagogue receptor (GHS-R) 1a (SB203580, SP600125, and [D-Lys3]-GHRP-6, respectively), were administered for seven consecutive days. Levels of various signaling molecules were assessed in isolated rat peritoneal macrophages. The results showed that serum ghrelin levels and levels of macrophage glucocorticoid receptor (GR) decreased, while phosphorylation of p38MAPK, JNK, and p65 nuclear factor (NF) κB increased. Ghrelin inhibited the serum induction of proinflammatory mediators, especially TNF-α, and promoted wound healing in a dose-dependent manner. Ghrelin treatment decreased phosphorylation of p38MAPK, JNK, and p65NF-κB, and increased GR levels in the presence of GHS-R1a. SB203580 or co-administration of SB203580 and SP600125 decreased TNF-α level, which may have contributed to the inactivation of p65NF-κB and increase in GR expression, as confirmed by western blotting. In conclusion, ghrelin enhances wound recovery in CRBI rats, possibly by decreasing the induction of TNF-α or other proinflammatory mediators that are involved in the regulation of GHS-R1a-mediated MAPK-NF-κB/GR signaling pathways.

  14. IL-1α/IL-1R1 expression in chronic obstructive pulmonary disease and mechanistic relevance to smoke-induced neutrophilia in mice.

    Fernando M Botelho

    Full Text Available BACKGROUND: Cigarette smoking is the main risk factor for the development of chronic obstructive pulmonary disease (COPD, a major cause of morbidity and mortality worldwide. Despite this, the cellular and molecular mechanisms that contribute to COPD pathogenesis are still poorly understood. METHODOLOGY AND PRINCIPAL FINDINGS: The objective of this study was to assess IL-1 α and β expression in COPD patients and to investigate their respective roles in perpetuating cigarette smoke-induced inflammation. Functional studies were pursued in smoke-exposed mice using gene-deficient animals, as well as blocking antibodies for IL-1α and β. Here, we demonstrate an underappreciated role for IL-1α expression in COPD. While a strong correlation existed between IL-1α and β levels in patients during stable disease and periods of exacerbation, neutrophilic inflammation was shown to be IL-1α-dependent, and IL-1β- and caspase-1-independent in a murine model of cigarette smoke exposure. As IL-1α was predominantly expressed by hematopoietic cells in COPD patients and in mice exposed to cigarette smoke, studies pursued in bone marrow chimeric mice demonstrated that the crosstalk between IL-1α+ hematopoietic cells and the IL-1R1+ epithelial cells regulates smoke-induced inflammation. IL-1α/IL-1R1-dependent activation of the airway epithelium also led to exacerbated inflammatory responses in H1N1 influenza virus infected smoke-exposed mice, a previously reported model of COPD exacerbation. CONCLUSIONS AND SIGNIFICANCE: This study provides compelling evidence that IL-1α is central to the initiation of smoke-induced neutrophilic inflammation and suggests that IL-1α/IL-1R1 targeted therapies may be relevant for limiting inflammation and exacerbations in COPD.

  15. Novel osmotin inhibits SREBP2 via the AdipoR1/AMPK/SIRT1 pathway to improve Alzheimer's disease neuropathological deficits

    Shah, S A; Yoon, G H; Chung, S S; Abid, M N; Kim, T H; Lee, H Y; Kim, M O


    Extensive evidence has indicated that a high rate of cholesterol biogenesis and abnormal neuronal energy metabolism play key roles in Alzheimer's disease (AD) pathogenesis. Here, for we believe the first time, we used osmotin, a plant protein homolog of mammalian adiponectin, to determine its therapeutic efficacy in different AD models. Our results reveal that osmotin treatment modulated adiponectin receptor 1 (AdipoR1), significantly induced AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) activation and reduced SREBP2 (sterol regulatory element-binding protein 2) expression in both in vitro and in vivo AD models and in Adipo−/− mice. Via the AdipoR1/AMPK/SIRT1/SREBP2 signaling pathway, osmotin significantly diminished amyloidogenic Aβ production, abundance and aggregation, accompanied by improved pre- and post-synaptic dysfunction, cognitive impairment, memory deficits and, most importantly, reversed the suppression of long-term potentiation in AD mice. Interestingly, AdipoR1, AMPK and SIRT1 silencing not only abolished osmotin capability but also further enhanced AD pathology by increasing SREBP2, amyloid precursor protein (APP) and β-secretase (BACE1) expression and the levels of toxic Aβ production. However, the opposite was true for SREBP2 when silenced using small interfering RNA in APPswe/ind-transfected SH-SY5Y cells. Similarly, osmotin treatment also enhanced the non-amyloidogenic pathway by activating the α-secretase gene that is, ADAM10, in an AMPK/SIRT1-dependent manner. These results suggest that osmotin or osmotin-based therapeutic agents might be potential candidates for AD treatment. PMID:27001618

  16. Differential Regulation of ERK1/2 and mTORC1 Through T1R1/T1R3 in MIN6 Cells

    Wauson, Eric M.; Guerra, Marcy L.; Dyachok, Julia; McGlynn, Kathleen; Giles, Jennifer; Ross, Elliott M.; Cobb, Melanie H.


    The MAPKs ERK1/2 respond to nutrients and other insulin secretagogues in pancreatic β-cells and mediate nutrient-dependent insulin gene transcription. Nutrients also stimulate the mechanistic target of rapamycin complex 1 (mTORC1) to regulate protein synthesis. We showed previously that activation of both ERK1/2 and mTORC1 in the MIN6 pancreatic β-cell-derived line by extracellular amino acids (AAs) is at least in part mediated by the heterodimeric T1R1/T1R3, a G protein-coupled receptor. We ...

  17. Determination of 63Ni and 59Ni in spent ion-exchange resin and activated charcoal from the IEA-R1 nuclear research reactor.

    Taddei, M H T; Macacini, J F; Vicente, R; Marumo, J T; Sakata, S K; Terremoto, L A A


    A radiochemical method has been adapted to determine (59)Ni and (63)Ni in samples of radioactive wastes from the water cleanup system of the IEA-R1 nuclear research reactor. The process includes extraction chromatographic resin with dimethylglyoxime (DMG) as a functional group. Activity concentrations of (59)Ni and (63)Ni were measured, respectively, by X-ray spectrometry and liquid scintillation counting, whereas the chemical yield was determined by ICP-OES. The average ratio of measured activity concentrations of (63)Ni and (59)Ni agree well with theory. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Validation of the βeta-s.t.a.r. 1+1 for rapid screening of residues of β-lactam antibiotics in milk

    Reybroeck, Wim; Ooghe, Sigrid; De Brabander, Hubert; Daeseleire, Els


    Abstract The a two minute protocol (1+1) for the ?eta-s.t.a.r., manufactured by Neogen Corporation, Lansing, MI, USA, was validated at the Technology and Food Science Unit of the Institute for Agricultural and Fisheries Research according to Commission Decision 2002/657/EC. The test was very selective for the group of ?-lactam compounds: the only interference found was by clavulanic acid at 2500 ?g kg-1 and above. The modified protocol (?eta-s.t.a.r. 1+1) detected all ?-lactams wit...

  19. Algunas representaciones de la función hipergeométrica generalizada 2R1 (a, b; c; τ ; z

    Jaime Castillo Pérez


    Full Text Available El campo de las funciones especiales ha tenido un gran desarrollo en los últimos decenios, dado que son muchos campos de aplicación de las mismas, como por ejemplo procesos estocásticos relacionados, investigación de operaciones, teoría cuántica, ecuaciones funcionales, vibración de placas, conducción del calor, elasticidad, radiación. En este trabajo se considera una ampliación de las teorías presentadas por M. Dotsenko en 1991, quien introdujo la generalización de la función hipergeométrica de Gauss denotada τ por 2 R1 (z, y estableció su representación en serie e integral. Es importante notar que en 1999 Nina Virchenko, y luego en el 2003 Leda Galué, consideraron esta función introduciendo un conjunto de fórmulas de recurrencia y de diferenciación. En este trabajo se establecen algunas representaciones para la función 2 R1 (a, b; c; τ ; z, las cuales serán muy útiles puesto que permiten simplificar cálculos a la hora de resolver problemas que involucren esta función.

  20. Expression of adiponectin receptors, AdipoR1 and AdipoR2, in normal colon epithelium and colon cancer tissue.

    Yoneda, Kyoko; Tomimoto, Ayako; Endo, Hiroki; Iida, Hiroshi; Sugiyama, Michiko; Takahashi, Hirokazu; Mawatari, Hironori; Nozaki, Yuichi; Fujita, Koji; Yoneda, Masato; Inamori, Masahiko; Nakajima, Noriko; Wada, Koichiro; Nagashima, Yoji; Nakagama, Hitoshi; Uozaki, Hitoshi; Fukayama, Masashi; Nakajima, Atsushi


    Adiponectin is secreted by adipocytes and is a key hormone responsible for insulin sensitization. Recent studies have shown that plasma adiponectin is decreased in patients with breast, endometrial and gastric cancer. However, the effect of adiponectin on colorectal carcinogenesis is controversial. It is now well known that the adiponectin receptor exists in two isoforms, adiponectin receptor 1 (AdipoR1) and adiponectin receptor 2 (AdipoR2). We examined the expression of the adiponectin receptors on normal colon mucosa and colon cancer tissues in a human study using real-time RT-PCR, Western blotting and immunohistochemical staining. Adiponectin receptors, AdipoR1/AdipoR2, were expressed in normal colon epithelial and colon cancer cells. Furthermore, laser microdissection was performed to confirm our results. These results suggest that adiponectin may exert some effects on normal colon epithelium or colon cancer cells directly through adiponectin receptors. Further studies are required to elucidate the function of the AdipoRs activated by adiponectin and the downstream mechanisms of AdipoRs in colon cancer cells.

  1. Faint moving object detection, and the Low Signal-to-Noise recovery of Main Belt comet P/2008 R1 Garradd

    Kleyna, Jan; Hainaut, Olivier


    We describe the recovery of faint Main Belt comet P/2008 R1 Garradd using several telescopes, culminating in a successful low $S/N$ recovery with the Gemini North telescope with GMOS. This recovery was a time-critical effort for a mission proposal, and had to be performed in a crowded field. We describe techniques and software tools for eliminating systematic noise artifacts and stellar residuals, bringing the final detection image statistics close to the Gaussian ideal for a median image stack, and achieving a detection sensitivity close to this theoretical optimum. The magnitude of $R_c$=26.1$\\pm$0.2 with an assumed geometric albedo of 0.05 corresponds to a radius of 0.3 km. For ice to have survived in this object over the age of the solar system, it implies that the object is a more recent collisional fragment. We discuss the implications of the unexpectedly faint magnitude and nuclear size of P/2008 R1 on the survival of ice inside very small bodies.

  2. Discovery of Western European R1b1a2 Y chromosome variants in 1000 genomes project data: an online community approach.

    Richard A Rocca

    Full Text Available The authors have used an online community approach, and tools that were readily available via the Internet, to discover genealogically and therefore phylogenetically relevant Y-chromosome polymorphisms within core haplogroup R1b1a2-L11/S127 (rs9786076. Presented here is the analysis of 135 unrelated L11 derived samples from the 1000 Genomes Project. We were able to discover new variants and build a much more complex phylogenetic relationship for L11 sub-clades. Many of the variants were further validated using PCR amplification and Sanger sequencing. The identification of these new variants will help further the understanding of population history including patrilineal migrations in Western and Central Europe where R1b1a2 is the most frequent haplogroup. The fine-grained phylogenetic tree we present here will also help to refine historical genetic dating studies. Our findings demonstrate the power of citizen science for analysis of whole genome sequence data.

  3. Synthesis and pharmacological evaluation of 2-(1-alkylpiperidin-4-yl)-n-[(1r)-1-(4-fluorophenyl)-2-methylpropyl]acetamide Derivatives as Novel Antihypertensive Agents.

    Watanuki, Susumu; Matsuura, Keisuke; Tomura, Yuichi; Okada, Minoru; Okazaki, Toshio; Ohta, Mitsuaki; Tsukamoto, Shin-ichi


    We synthesized and evaluated the inhibitory activity of a series of 2-(1-alkylpiperidin-4-yl)-N-[(1R)-1-(4-fluorophenyl)-2-methylpropyl]acetamide derivatives against T-type Ca(2+) channels. Structure-activity relationship studies revealed that the position of the amide structure was important for the potent inhibitory activity toward T-type Ca(2+) channels. In addition, the introduction of an appropriate substituent on the pendant benzene ring played a crucial role for the selectivity towards T-type Ca(2+) channels over L-type Ca(2+) channels and the potent bradycardic activity of these derivatives. Oral administration of N-[(1R)-1-(4-fluorophenyl)-2-methylpropyl]-2-(1-{2-[2-(2-methoxyethoxy)phenyl]ethyl}piperidin-4-yl)acetamide (4f), which had superior selectivity for T-type Ca(2+) channels over L-type Ca(2+) channels, lowered blood pressure in spontaneously hypertensive rats without inducing reflex tachycardia, which is often caused by traditional L-type Ca(2+) channel blockers.

  4. The Production of Benzaldehyde by Rhizopus oligosporus USM R1 in a Solid State Fermentation (SSF System of Soy Bean Meal: Rice Husks

    Norliza, A. W.


    Full Text Available The cultivation of Rhizopus oligosporus USM R1 for the production of benzaldehyde, a bitter cherry almond flavour was performed using soya bean meal and rice husks as the substrates. The identification of R. oligosporus USM R1 was performed based on the observation made under light microscope and scanning electron microscope (SEM. The optimum conditions for the SSF in a 250-ml Erlenmeyer flask system were 40% (v/w water content, substrate particle size of 0.7 mm; inoculum size of 1 x 10^5 spores/g substrate; incubation temperature of 30C; substrate amount of 7 g and the ratio of soy bean meal: rice husks of 50:50%. A maximum benzaldehyde production was obtained when the substrate was agitated after 48 hour for a 96 hour fermentation time. The highest benzaldehyde production obtained after 96 hour cultivation was 5.47 mg g-1 substrate. The supplementation of carbon and nitrogen sources in the substrate mixture revealed an enhancement in the growth and benzyldehyde production. A maximum production of benzaldehyde was obtained with the supplementation of L-phenylalanine, a precursor for benzaldehyde biosynthesis which gave 38.69 mg benzaldehyde/g substrate. This is approximately 6-folds higher compared to the substrates without the supplementation of L-phenylalanine.

  5. 早期母婴分离对大鼠成年后抑郁易感性和海马GluR1表达的影响%Effects of maternal separation on vulnerability to depression and the expression of GluR1 in adult rats

    黄铭发; 许婷; 危智盛; 李旭然; 郑玉; 周芩稷; 董建阳; 蔡翔


    Objective To investigate the effect of maternal separation on vulnerability to depression and the ex-pression of GluR1 in adult rats .Methods Fifty-six newborn male SD rats were randomly assigned to four groups , con-trol, MS, CRR and MS+CRR groups.The rats in MS and MS+CRR groups received maternal separated for 3 h on Day 2 to Day 14 after birth.The rats in CRR and MS+CRR groups were subjected to three -hour chronic repeated restraint stress ( CRR) for 10 days at 6 weeks old ( weight were sub behavioral changes were recorded by open -field test ( OFT) , novelty suppressed feeding test ( NST) and forced swim test ( FST) .Western blot was used to measure the expression of GluR1 in hippocampus .Results Compared with the control , MS and CRR groups , the total distance of OFT in MS +CRR group was significantly reduced , with significant increases of the latency time of NSF and the immobility time of FST (P<0.05).The expression of GluR1 in hippocampus in MS group was down -regulatedcompared to control group (P<0.05).Compared with the other three groups , significant down-regulation in GluR1 in hippocampuswas revealed in MS+CRR group (P<0.05).Conclusion Maternal separation causes vulnerability to depression in rats at adult age .And the expression of GluR1 in hippocampus is involved .%目的:探讨早期母婴分离对大鼠成年后抑郁易感性和海马GluR1表达的影响。方法将Sprague Dawley孕鼠分娩的56只新生雄性幼鼠随机分为对照组、母婴分离(MS)组、慢性重复性束缚应激(CRR)组和母婴分离并慢性重复性束缚应激( MS+CRR)组。 MS组和MS+CRR组幼鼠出生后第2~14天进行母婴分离应激3 h。鼠龄达7周(体重≈290 g)后,CRR组和MS+CRR组大鼠连续10 d给予3 h慢性重复性束缚应激。之后,进行旷场、新环境进食抑制及强迫游泳实验。结果与对照组、MS组和CRR组比较,MS+CRR组旷场总距离明显减少,新环境进食抑制实验进食潜

  6. Role of metabotropic glutamate receptor subtype mGluR1 in brief nociception and central sensitization of primate STT cells.

    Neugebauer, V; Chen, P S; Willis, W D


    G-protein coupled metabotropic glutamate receptors (mGluRs) are important modulators of synaptic transmission in the mammalian CNS and have been implicated in various forms of neuroplasticity and nervous system disorders. Increasing evidence also suggests an involvement of mGluRs in nociception and pain behavior although the contribution of individual mGluR subtypes is not yet clear. Subtypes mGluR1 and mGluR5 are classified as group I mGluRs and share the ability to stimulate phosphoinositide hydrolysis and activate protein kinase C. The present study examined the role of group I mGluRs in nociceptive processing and capsaicin-induced central sensitization of primate spinothalamic tract (STT) cells in vivo. In 10 anesthetized male monkeys (Macaca fascicularis) extracellular recordings were made from 20 STT cells in the lumbar dorsal horn. Responses to brief (15 s) cutaneous stimuli of innocuous (BRUSH) and barely and substantially noxious (PRESS and PINCH, respectively) intensity were recorded before, during, and after the infusion of group I mGluR agonists and antagonists into the dorsal horn by microdialysis. Cumulative concentration-response relationships were obtained by applying different concentrations for at least 20 min each (at 5 microl/min). The actual concentrations reached in the tissue are 2-3 orders of magnitude lower than those in the microdialysis fibers (values in this paper refer to the latter). The group I antagonists were also applied at 10-25 min after capsaicin injection. S-DHPG, a group I agonist at both mGluR1 and mGluR5, potentiated the responses to innocuous and noxious stimuli (BRUSH > PRESS > PINCH) at low concentrations (10-100 microM; n = 5) but had inhibitory effects at higher concentrations (1-10 mM; n = 5). The mGluR5 agonist CHPG (1 microM-100 mM; n = 5) did not potentiate but inhibited all responses (10-100 mM; n = 5). AIDA (1 microM-100 mM), a mGluR1-selective antagonist, dose-dependently depressed the responses to PINCH and

  7. 冠心病患者血清同型半胱氨酸、miR-1、miR-126和 miR-208的相关性分析%Correlation analysis of Hcy,miR-1,miR-126 and miR-208 in coronary heart disease patients

    金洁; 王俊; 王磊; 蔡晓敏; 王立军; 宫剑滨


    Objective To study the correlation between miRNA (miR-1,miR-126,miR-208) and homocysteine (Hcy) con-bining on the severity of stenosis of coronary artery .Methods One hundred and two patients undergoing quantitative coronary angiogra -phy were involved in this study .Patients were categorized into control group and coronary artery disease ( CAD) group.The concentra-tion of Hcy and miRNA (miR-1,miR-126,miR-208) were measured.The severity scale of coronary artery stenosis was quantitatively assessed according to coronary angiography by Gensini score .The correlation analysis and linear regression were used to show the asso-ciation of Hcy,miRNA (miR-1,miR-126 and miR-208) and Gensini score.Results The concentration of Hcy and miRNA (miR-1, miR-126 and miR-208) were higher in CAD group than those in control group .In the correlation analysis ,Gensini score was positively associated with Hcy,miR-1 and miR-126 (r=0.575,0.649,and 0.499,P<0.01) and the positive correlation between them remained significant after revising partial correlation analysis (r'=0.693,0.621,and 0.532,P<0.05).Hcy was influenced by miR-1 and miR-126 both in the Spearson correlation analysis (r=0.513,0.56,P<0.01) and partial correlation analysis (r'=0.599,0.614,P<0.05),but not by miR-208.Conclusion Hcy,miRNA (miR-1,miR-126 and miR-208) are independent risk factor for coronary heart disease and are increased along with stenosis of coronary arteries .%目的:结合冠状动脉狭窄程度,探讨冠心病患者血清同型半胱氨酸( Hcy)与微小RNA( miR-1、miR-126及miR-208)之间的相互关系。方法选取行冠脉造影术的患者共102名,根据造影结果分为对照组和观察组,检测Hcy、miR-1、miR-126及miR-208水平,同时对冠脉造影结果进行评价并计算Gensini积分,应用相关及线性回归分析Hcy、miRNA( miR-1、miR-126、miR-208)以及Gensini积分的相互关系。结果观察组血清Hcy、miRNA(miR-1、miR-126、miR-208)水平均

  8. Ryanodine receptor type 1 (RyR1) mutations C4958S and C4961S reveal excitation-coupled calcium entry (ECCE) is independent of sarcoplasmic reticulum store depletion.

    Hurne, Alanna M; O'Brien, Jennifer J; Wingrove, Douglas; Cherednichenko, Gennady; Allen, Paul D; Beam, Kurt G; Pessah, Isaac N


    Bi-directional signaling between ryanodine receptor type 1 (RyR1) and dihydropyridine receptor (DHPR) in skeletal muscle serves as a prominent example of conformational coupling. Evidence for a physiological mechanism that upon depolarization of myotubes tightly couples three calcium channels, DHPR, RyR1, and a Ca(2+) entry channel with SOCC-like properties, has recently been presented. This form of conformational coupling, termed excitation-coupled calcium entry (ECCE) is triggered by the alpha(1s)-DHPR voltage sensor and is highly dependent on RyR1 conformation. In this report, we substitute RyR1 cysteines 4958 or 4961 within the TXCFICG motif, common to all ER/SR Ca(2+) channels, with serine. When expressed in skeletal myotubes, C4958S- and C4961S-RyR1 properly target and restore L-type current via the DHPR. However, these mutants do not respond to RyR activators and do not support skeletal type EC coupling. Nonetheless, depolarization of cells expressing C4958S- or C4961S-RyR1 triggers calcium entry via ECCE that resembles that for wild-type RyR1, except for substantially slowed inactivation and deactivation kinetics. ECCE in these cells is completely independent of store depletion, displays a cation selectivity of Ca(2+)>Sr(2+) approximately Ba(2+), and is fully inhibited by SKF-96365 or 2-APB. Mutation of other non-CXXC motif cysteines within the RyR1 transmembrane assembly (C3635S, C4876S, and C4882S) did not replicate the phenotype observed with C4958S- and C4961S-RyR1. This study demonstrates the essential role of Cys(4958) and Cys(4961) within an invariant CXXC motif for stabilizing conformations of RyR1 that influence both its function as a release channel and its interaction with ECCE channels.

  9. miR-1 suppresses the growth of esophageal squamous cell carcinoma in vivo and in vitro through the downregulation of MET, cyclin D1 and CDK4 expression



    Several aberrant microRNAs (miRNAs or miRs) have been implicated in esophageal cancer (EC), which is widely prevalent in China. However, their role in EC tumorigenesis has not yet been fully elucidated. In the present study, we determined that miR-1 was downregulated in esophageal squamous cell carcinoma (ESCC) tissues compared with adjacent non-neoplastic tissues using RT-qPCR, and confirmed this using an ESCC cell line. Using a nude mouse xenograft model, we confirmed that the re-expression of miR-1 significantly inhibited ESCC tumor growth. A tetrazolium assay and a trypan blue exclusion assay revealed that miR-1 suppressed ESCC cell proliferation and increased apoptosis, whereas the silencing of miR-1 promoted cell proliferation and decreased apoptosis, suggesting that miR-1 is a novel tumor suppressor. To elucidate the molecular mechanisms of action of miR-1 in ESCC, we investigated putative targets using bioinformatics tools. MET, cyclin D1 and cyclin-dependent kinase 4 (CDK4), which are involved in the hepatocyte growth factor (HGF)/MET signaling pathway, were found to be targets of miR-1. miR-1 expression inversely correlated with MET, cyclin D1 and CDK4 expression in ESCC cells. miR-1 directly targeted MET, cyclin D1 and CDK4, suppressing ESCC cell growth. The newly identified miR-1/MET/cyclin D1/CDK4 axis provides new insight into the molecular mechanisms of ESCC pathogenesis and indicates a novel strategy for the diagnosis and treatment of ESCC. PMID:27247259

  10. Pain-related increase of excitatory transmission and decrease of inhibitory transmission in the central nucleus of the amygdala are mediated by mGluR1

    Neugebauer Volker


    Full Text Available Abstract Neuroplasticity in the central nucleus of the amygdala (CeA, particularly its latero-capsular division (CeLC, is an important contributor to the emotional-affective aspects of pain. Previous studies showed synaptic plasticity of excitatory transmission to the CeLC in different pain models, but pain-related changes of inhibitory transmission remain to be determined. The CeLC receives convergent excitatory inputs from the parabrachial nucleus in the brainstem and from the basolateral amygdala (BLA. In addition, feedforward inhibition of CeA neurons is driven by glutamatergic projections from the BLA area to a cluster of GABAergic neurons in the intercalated cell masses (ITC. Using patch-clamp in rat brain slices we measured monosynaptic excitatory postsynaptic currents (EPSCs and polysynaptic inhibitory currents (IPSCs that were evoked by electrical stimulation in the BLA. In brain slices from arthritic rats, input-output functions of excitatory synaptic transmission were enhanced whereas inhibitory synaptic transmission was decreased compared to control slices from normal untreated rats. A non-NMDA receptor antagonist (NBQX blocked the EPSCs and reduced the IPSCs, suggesting that non-NMDA receptors mediate excitatory transmission and also contribute to glutamate-driven feed-forward inhibition of CeLC neurons. IPSCs were blocked by a GABAA receptor antagonist (bicuculline. Bicuculline increased EPSCs under normal conditions but not in slices from arthritic rats, which indicates a loss of GABAergic control of excitatory transmission. A metabotropic glutamate receptor subtype 1 (mGluR1 antagonist (LY367385 reversed both the increase of excitatory transmission and the decrease of inhibitory transmission in the arthritis pain model but had no effect on basal synaptic transmission in control slices from normal rats. The inhibitory effect of LY367385 on excitatory transmission was blocked by bicuculline suggesting the involvement of a GABAergic

  11. CCD polarimetry of distant comets C/2010 S1 (LINEAR) and C/2010 R1 (LINEAR) at the 6-m telescope of the SAO RAS

    Ivanova, Oleksandra V.; Dlugach, Janna M.; Afanasiev, Viktor L.; Reshetnyk, Volodymyr M.; Korsun, Pavlo P.


    We present first measurements of the degree of linear polarization of distant comets C/2010 S1 (LINEAR) and C/2010 R1 (LINEAR) at heliocentric distances r=5.9-7.0 AU. Observations were carried out with the SCORPIO-2 focal reducer at the 6-m telescope of the Special Astrophysical Observatory (Russia). Both comets showed considerable level of activity (significant dust comae and tails) beyond a zone where water ice sublimation is negligible (up to 5 AU). Significant spatial variations both in the intensity and polarization are found in both the comets. The slope of radial profiles of intensity changes gradually with the distance from the photocenter: from -0.7 near the nucleus up to about -1.3 for larger distances (up to 100,000 km). The variation in polarization profiles indicates the non-uniformity in the polarization distribution over the coma. The polarization degree over the coma gradually increases (in absolute value) with increasing photocentric distance from of about -1.9% up to -3% for comet C/2010 S1 (LINEAR), and from of about -2.5% up to -3.5% for comet C/2010 R1 (LINEAR). These polarization values are significantly higher than typical value of the whole coma polarization (∼-1.5%) for comets at heliocentric distances less than 5 AU. The obtained photometric and polarimetric data are compared with those derived early for other comets at smaller heliocentric distances. Numerical modeling of light scattering characteristics was performed for media composed of particles with different refractive indexes, shapes, and sizes. The computations were made by using the superposition T-matrix method. We obtained that for comet C/2010 S1 (LINEAR), the dust in the form of aggregates of overall radius R~1.3 μm composed of N=1000 spherical monomers with radius a=0.1 μm and refractive index m=1.65+i0.05, allows to obtain a satisfactory agreement between the results of polarimetric observations of comet C/2010 S1 and computations.

  12. Efficient anti-Prelog enantioselective reduction of acetyltrimethylsilane to (R-1-trimethylsilylethanol by immobilized Candida parapsilosis CCTCC M203011 cells in ionic liquid-based biphasic systems

    Zhang Bo-Bo


    Full Text Available Abstract Background Biocatalytic asymmetric reductions with whole cells can offer high enantioselectivity, environmentally benign processes and energy-effective operations and thus are of great interest. The application of whole cell-mediated bioreduction is often restricted if substrate and product have low water solubility and/or high toxicity to the biocatalyst. Many studies have shown that a biphasic system is often useful in this instance. Hence, we developed efficient biphasic reaction systems with biocompatible water-immiscible ionic liquids (ILs, to improve the biocatalytic anti-Prelog enantioselective reduction of acetyltrimethylsilane (ATMS to (R-1-trimethylsilylethanol {(R-1-TMSE}, which is key synthon for a large number of silicon-containing drugs, using immobilized Candida parapsilosis CCTCC M203011 cells as the biocatalyst. Results It was found that the substrate ATMS and the product 1-TMSE exerted pronounced toxicity to immobilized Candida parapsilosis CCTCC M203011 cells. The biocompatible water-immiscible ILs can be applied as a substrate reservoir and in situ extractant for the product, thus greatly enhancing the efficiency of the biocatalytic process and the operational stability of the cells as compared to the IL-free aqueous system. Various ILs exerted significant but different effects on the bioreduction and the performances of biocatalysts were closely related to the kinds and combination of cation and anion of ILs. Among all the water-immiscible ILs investigated, the best results were observed in 1-butyl-3-methylimidazolium hexafluorophosphate (C4mim·PF6/buffer biphasic system. Furthermore, it was shown that the optimum substrate concentration, volume ratio of buffer to IL, buffer pH, reaction temperature and shaking rate for the bioreduction were 120 mM, 8/1 (v/v, 6.0, 30°C and 180 r/min, respectively. Under these optimized conditions, the initial reaction rate, the maximum yield and the product e.e. were 8.1

  13. 孕激素对子宫内膜癌Ishikawa细胞TGF-β1、TβR1表达的影响%Effects of Progesterone on TGF-β1 and TβR1 in Ishikawa Cell of Endometrial Carcinoma

    王改华; 刘贵鹏


    Objective Toinvestigate the effects of progesterone(MPA)onapoptosis and TGFβ1、TβR1 protein expression of Ishikawa cell line of human endometrial carcinoma in vitro. Methods The proliferation of Ishikawa cells in vitro was determined by methyl thiazolyl terazolium (MTT) method. Percentage of cell apoptosis was detected by flow cytometry (FCM). The expression of TGF-β1、TβR1 trotein of Ishikawa cells was detected by streptavidin-peroxidase linkage (SP) method. Results Progesterone inhibited the growth of Ishikawa cells significantly when the concentration of progesterone reached 15 μg/ml (P < 0.01 ). Flow cytometry (FCM) showed that the apoptotic rate in progesterone-treated group was sgnificantly increased (P < 0.05) compared with control group. Apotosis morphology was observed in each group. The expression of TGF-β1 in Ishikawa cells was decreased after the cells had been treated with MPA for 72 hours,while expression of TβR1 was increased. Conclusion Progesterone inhibited the proliferation and induced the apoptosis ofIshikawa cells. The regulation on the expressions ofthe TGF-β1 and TβR1 proteins by progesterone seems to be responsible for the effects on the apoptosis of Ishikawa cells.%目的 观察孕酮(MPA)对人子宫内膜高分化腺癌细胞株Ishikawa细胞中TGF-β1、TβR1蛋白表达的影响,探讨孕激素治疗子宫内膜腺癌的作用机制.方法 体外培养人子宫内膜高分化腺癌细胞株Ishikawa细胞,MTT法观察细胞的增殖;流式细胞仪(FCM)检测细胞细胞凋亡率;链霉素抗生物素蛋白-过氧化物酶连接(SP)法测定Ishikawa细胞中TGF-β1、TβR1蛋白的表达.结果 MPA呈时间及剂量依赖性抑制Ishikawa细胞增殖(P<0.05).与对照组相比,MPA处理组细胞凋亡率上升(P<0.01).TGF-β1在Ishikawa细胞中有强表达,MPA处理72 h后,其表达明显下调(P<0.01);TβR1蛋白在Ishikawa细胞中有弱表达,MPA处理72 h后,明显上调其表达(P<0.01).结论 孕酮调节的TCFβ1、TβR

  14. Crystal structure of 2-{(R)-[1-(4-bromo-phen-yl)eth-yl]imino-meth-yl}-4-(phenyl-diazen-yl)phenol, a chiral photochromic Schiff base.

    Moriwaki, Ryoji; Akitsu, Takashiro


    The title chiral photochromic Schiff base compound, C21H18BrN3O, was synthesized from (R)-(+)-1-(4-bromo-phen-yl)ethyl-amine and the salicyl-aldehyde of an azo-benzene derivative. The mol-ecule corresponds to the phenol-imine tautomer, the C=N and N-C bond distances being 1.285 (3) and 1.470 (3) Å, respectively. The diazenyl group adopts a trans form, with an N=N distance of 1.256 (3) Å. The hy-droxy group is involved in intra-molecular O-H⋯N hydrogen bonding. In the crystal, C-H⋯π inter-actions consolidate the crystal packing of one-dimensional chains, which exhibits short inter-molecular Br⋯C contacts of 3.400 (3) Å.

  15. Identification of the chromosome complement and the spontaneous 1R/1V translocations in allotetraploid Secale cereale × Dasypyrum villosum hybrids through cytogenetic approaches.

    Książczyk, Tomasz; Apolinarska, Barbara; Kulak-Książczyk, Sylwia; Wiśniewska, Halina; Stojałowski, Stefan; Łapiński, Mirosław


    Genome modifications that occur at the initial interspecific hybridization event are dynamic and can be consolidated during the process of stabilization in successive generations of allopolyploids. This study identifies the number and chromosomal location of ribosomal DNA (rDNA) sites between Secale cereale, Dasypyrum villosum, and their allotetraploid S. cereale × D. villosum hybrids. For the first time, we show the advantages of FISH to reveal chromosome rearrangements in the tetraploid Secale × Dasypyrum hybrids. Based on the specific hybridization patterns of ribosomal 5S, 35S DNA and rye species-specific pSc200 DNA probes, a set of genotypes with numerous Secale/Dasypyrum translocations of 1R/1V chromosomes were identified in successive generations of allotetraploid S. cereale × D. villosum hybrids. In addition we analyse rye chromosome pairs using FISH with chromosome-specific DNA sequences on S. cereale × D. villosum hybrids.

  16. Neuroprotective effect of β-asarone against Alzheimer’s disease: regulation of synaptic plasticity by increased expression of SYP and GluR1

    Liu SJ


    Full Text Available Si-jun Liu,1,* Cong Yang,2,* Yue Zhang,2,* Ru-yu Su,2 Jun-li Chen,2 Meng-meng Jiao,2 Hui-fang Chen,2 Na Zheng,2 Si Luo,2 Yun-bo Chen,2 Shi-jian Quan,1 Qi Wang21School of Chinese Materia Medica, 2Institute of Clinical Pharmacology, Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of China*These authors contributed equally to this workAim: β-asarone, an active component of Acori graminei rhizome, has been reported to have neuroprotective effects in Alzheimer’s disease. As the underlying mechanism is not known, we investigated the neuroprotective effects of β-asarone in an APP/PS1 double transgenic mouse model and in NG108 cells.Materials and methods: APPswe/PS1dE9 double transgenic male mice were randomly assigned to a model group, β-asarone treatment groups (21.2, 42.4, or 84.8 mg/kg/d, or donepezil treatment group (2 mg/kg/d. Donepezil treatment was a positive control, and background- and age-matched wild-type B6 mice were an external control group. β-asarone (95.6% purity was dissolved in 0.8% Tween 80 and administered by gavage once daily for 2.5 months. Control and model animals received an equal volume of vehicle. After 2.5 months of treatment, behavior of all animals was evaluated in a Morris water maze. Expression of synaptophysin (SYP and glutamatergic receptor 1 (G1uR1 in the hippocampus and cortex of the double transgenic mice was assayed by Western blotting. The antagonistic effects of β-asarone against amyloid-β peptide (Aβ were investigated in vitro in the NG108-15 cell line. After 24 hours of incubation, cells were treated with 10 µm Aβ with or without β-asarone at different concentrations (6.25, 12.5, or 25 µM for an additional 36 hours. The cytotoxicity of β-asarone was evaluated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assay of cell viability, and cell morphology was evaluated by bright-field microscopy after 24 hours of treatment. The expression of SYP and GluR1 in


    Yu. N. Kucheryavyi


    Full Text Available It is known that drugs based on the 1,2,4-triazole have a wide spectrum of biological activities. They are effective cardioprotectors, hepatoprotectors, anti-oxidants, which are widely used in cardiology practice. So this class of heterocyclic compounds has all the attributes of relevance. We have synthesized new 2-(5-(phenoxymethyl-4-R1-1,2,4-triazol-3-ylthio acetate acid and its salts. We have used complex physical and chemical methods of analysis to establish the structure of the synthesized compounds. The chemical properties and diuretic activity of synthesized compounds have been studied. The relation between the structure of obtained salts and their biological effect has been set. Compounds with strong diuretic activity have been found among the newly synthesized compounds. This fact testifies to further study and introduction into medical practice as the original drugs.

  18. Synthesis and Crystal Structure of 7α-[(R)-1-Hydroxyl-1-methyl-3-(thien-2-yl)-propyl]-6,14-endoethanotetrahydrothebaine

    LIU He; LIU Chun-He; WU Bo; CHEN Lan-Fu; ZHONG Bo-Hua


    The crystal structure of 7α-[(R)-1-hydroxyl-1-methyl-3-(thien-2-yl)propyl]-6,14-endoethanotetrahydro thebaine (C29H37NO4S, Mr = 495.66) has been determined by single-crystal X-ray diffraction analysis. The crystal belongs orthorhombic, space group P212121 with a = 10.063(4), b = 12.571(5), c = 19.626(8) (A), V = 2482.9(17) (A)3, Z = 4, Dc = 1.326 g/cm3,μ = 0.167 mm-1, F(000) = 1064, the final R = 0.0537 and wR = 0.0888. The C(7) substituent is 1-hydroxyl-1-methyl-3-(thienyl-2-)propyl group adopting R-configuration. The N-methyl group is located at the equatorial position as expected. The crystal structure shows the presence of intra-molecular hydrogen bonds,O(1)-H(1 )…O(2).

  19. Capture of influenza by medullary dendritic cells via SIGN-R1 is essential for humoral immunity in draining lymph nodes

    Gonzalez, Santiago F.; Lukacs-Kornek, Veronika; Kuligowski, Michael P.


    A major pathway for B cell acquisition of lymph-borne particulate antigens relies on antigen capture by subcapsular sinus macrophages of the lymph node. Here we tested whether this mechanism is also important for humoral immunity to inactivated influenza virus. By multiple approaches, including...... multiphoton intravital imaging, we found that antigen capture by sinus-lining macrophages was important for limiting the systemic spread of virus but not for the generation of influenza-specific humoral immunity. Instead, we found that dendritic cells residing in the lymph node medulla use the lectin receptor...... SIGN-R1 to capture lymph-borne influenza virus and promote humoral immunity. Thus, our results have important implications for the generation of durable humoral immunity to viral pathogens through vaccination....

  20. High-resolution spectroscopy of the CN red system in comet C/2013 R1 (Lovejoy) using WINERED at Koyama Astronomical Observatory

    Kawakita, Hideyo; Shinnaka, Yoshiharu; Kondo, Sohei; Hamano, Satoshi; Sameshima, Hiroaki; Nakanishi, Kenshi; Kawanishi, Takafumi; Nakaoka, Tetsuya; Otsubo, Shogo; Kinoshita, Masaomi; Ikeda, Yuji; Yamamoto, Ryo; Izumi, Natsuko; Fukue, Kei; Yasui, Chikako; Mito, Hiroyuki; Sarugaku, Yuki; Matsunaga, Noriyuki; Kobayashi, Naoto


    CN radical has the strong electronic transition moments in optical wavelength region and CN has extensively observed in comets. Especially, the CN violet system (B2Σ+—X2Σ+) has been observed by using high-resolution spectroscopic technique in order to infer the isotopic ratios of carbon and nitrogen in comets via 12C14N, 13C14N and 12C15N. However, the wavelength range for this system (~388 nm) is severely extinct if a comet is close to the Sun (we have to observe the comet at low elevations from the ground-based observatories). On the other hand, CN radical also has the strong electronic transition in near-infrared (~1.1 microns), the CN red system (A2Πi—X2Σ+). Although there are few reports on the high-resolution spectra of this band in comets, this wavelength region is not severely affected by the telluric extinction and considered as the new window for the observations of the carbon and nitrogen isotopic ratios in comets.High resolution near-infrared spectra of comet C/2013 R1 (Lovejoy) using the WINERED (R~3x104) spectrometer mounted on the 1.3-m Araki telescope at Koyama Astronomical Observatory were acquired on UT 2013 Nov 30. The heliocentric and geocentric distances were 0.91 AU and 0.49 AU, respectively. We detected strong emission lines of the CN red system (0,0) at around 1.1 microns. The rotational line intensities of this band approximately follow the Boltzmann distribution at ~300K for our observations. We present the detailed analysis of the CN red system in comet C/2013 R1 (Lovejoy) and discuss about the isotopic ratios in CN.This research program is supported by the MEXT --- Supported Program for the Strategic Research Foundation at Private Universities, 2014 - 2018 and partially supported by JSPS, 15J10864.

  1. Oxygen/glucose deprivation induces a reduction in synaptic AMPA receptors on hippocampal CA3 neurons mediated by mGluR1 and adenosine A3 receptors.

    Dennis, Siobhan H; Jaafari, Nadia; Cimarosti, Helena; Hanley, Jonathan G; Henley, Jeremy M; Mellor, Jack R


    Hippocampal CA1 pyramidal neurons are highly sensitive to ischemic damage, whereas neighboring CA3 pyramidal neurons are less susceptible. It is proposed that switching of AMPA receptor (AMPAR) subunits on CA1 neurons during an in vitro model of ischemia, oxygen/glucose deprivation (OGD), leads to an enhanced permeability of AMPARs to Ca(2+), resulting in delayed cell death. However, it is unclear whether the same mechanisms exist in CA3 neurons and whether this underlies the differential sensitivity to ischemia. Here, we investigated the consequences of OGD for AMPAR function in CA3 neurons using electrophysiological recordings in rat hippocampal slices. Following a 15 min OGD protocol, a substantial depression of AMPAR-mediated synaptic transmission was observed at CA3 associational/commissural and mossy fiber synapses but not CA1 Schaffer collateral synapses. The depression of synaptic transmission following OGD was prevented by metabotropic glutamate receptor 1 (mGluR1) or A(3) receptor antagonists, indicating a role for both glutamate and adenosine release. Inhibition of PLC, PKC, or chelation of intracellular Ca(2+) also prevented the depression of synaptic transmission. Inclusion of peptides to interrupt the interaction between GluA2 and PICK1 or dynamin and amphiphysin prevented the depression of transmission, suggesting a dynamin and PICK1-dependent internalization of AMPARs after OGD. We also show that a reduction in surface and total AMPAR protein levels after OGD was prevented by mGluR1 or A(3) receptor antagonists, indicating that AMPARs are degraded following internalization. Thus, we describe a novel mechanism for the removal of AMPARs in CA3 pyramidal neurons following OGD that has the potential to reduce excitotoxicity and promote neuroprotection.

  2. Oxygen/glucose Deprivation Induces a Reduction in Synaptic AMPA Receptors on Hippocampal CA3 Neurons Mediated by mGluR1 and A3 Receptors

    Dennis, Siobhan H.; Jaafari, Nadia; Cimarosti, Helena; Hanley, Jonathan G.; Henley, Jeremy M.; Mellor, Jack R.


    Summary Hippocampal CA1 pyramidal neurons are highly sensitive to ischemic damage, whereas neighbouring CA3 pyramidal neurons are less susceptible. It is proposed that switching of AMPA receptor (AMPAR) subunits on CA1 neurons during an in vitro model of ischemia, oxygen/glucose deprivation (OGD), leads to an enhanced permeability of AMPARs to Ca2+ resulting in delayed cell death. However, it is unclear if the same mechanisms exist in CA3 neurons and whether this underlies the differential sensitivity to ischemia. Here, we investigated the consequences of OGD for AMPAR function in CA3 neurons using electrophysiological recordings in rat hippocampal slices. Following a 15 minute OGD protocol a substantial depression of AMPAR-mediated synaptic transmission was observed at CA3 associational/commissural and mossy fiber synapses but not CA1 Schaffer collateral synapses. The depression of synaptic transmission following OGD was prevented by mGluR1 or A3 receptor antagonists, indicating a role for both glutamate and adenosine release. Inhibition of PLC, PKC or chelation of intracellular Ca2+ also prevented the depression of synaptic transmission. Inclusion of peptides to interrupt the interaction between GluA2 and PICK1 or dynamin and amphiphysin prevented the depression of transmission, suggesting a dynamin and PICK1-dependent internalisation of AMPARs after OGD. We also show a reduction in surface and total AMPAR protein levels after OGD was prevented by mGluR1 or A3 receptor antagonists indicating that AMPARs are degraded following internalisation. Thus, we describe a novel mechanism for the removal of AMPARs in CA3 pyramidal neurons following OGD that has the potential to reduce excitotoxicity and promote neuroprotection. PMID:21849555

  3. mGluR1 receptors contribute to non-purinergic slow excitatory transmission to submucosal VIP neurons of guinea-pig ileum

    Jaime Pei Pei Foong


    Full Text Available Vasoactive intestinal peptide (VIP immunoreactive secretomotor neurons in the submucous plexus are involved in mediating bacterial toxin-induced hypersecretion leading to diarrhoea. VIP neurons become hyperexcitable after the mucosa is exposed to cholera toxin, which suggests that the manipulation of the excitability of these neurons may be therapeutic. This study used standard intracellular recording methods to systematically characterize slow excitatory postsynaptic potentials (EPSPs evoked in submucosal VIP neurons by different stimulus regimes (1, 3 and 15 pulse 30 Hz stimulation, together with their associated input resistances and pharmacology. All slow EPSPs were associated with a significant increase in input resistance compared to baseline values. Slow EPSPs evoked by a single stimulus were confirmed to be purinergic, however, slow EPSPs evoked by 15 pulse trains were non-purinergic and those evoked by 3 pulse trains were mixed. NK1 or NK3 receptor antagonists did not affect slow EPSPs. The group I mGluR receptor antagonist, PHCCC reduced the amplitude of purinergic and non-purinergic slow EPSPs. Blocking mGluR1 receptors depressed the overall response to 3 and 15 pulse trains, but this effect was inconsistent, while blockade of mGluR5 receptors had no effect on the non-purinergic slow EPSPs. Thus, although other receptors are almost certainly involved, our data indicate that there are at least two pharmacologically distinct types of slow EPSPs in the VIP secretomotor neurons: one mediated by P2Y receptors and the other in part by mGluR1 receptors.

  4. Magnetic thermoablation stimuli alter BCL2 and FGF-R1 but not HSP70 expression profiles in BT474 breast tumors

    Stapf M


    Full Text Available Marcus Stapf, Nadine Pömpner, Melanie Kettering, Ingrid Hilger Institute of Diagnostic and Interventional Radiology, Jena University Hospital, Jena, Germany Abstract: Magnetically induced heating of magnetic nanoparticles (MNP in an alternating magnetic field (AMF is a promising minimal invasive tool for localized tumor treatment that eradicates tumor cells by applying thermal stress. While temperatures between 42°C and 45°C induce apoptosis and sensitize the cells for chemo- and radiation therapies when applied for at least 30 minutes, temperatures above 50°C, so-called thermoablative temperatures, rapidly induce irreversible cell damage resulting in necrosis. Since only little is known concerning the protein expression of anti-apoptotic B-cell lymphoma 2 (BCL2, fibroblast growth factor receptor 1 (FGF-R1, and heat shock protein (HSP70 after short-time magnetic thermoablative tumor treatment, these relevant tumor proteins were investigated by immunohistochemistry (IHC in a human BT474 breast cancer mouse xenograft model. In the investigated sample groups, the application of thermoablative temperatures (<2 minutes led to a downregulation of BCL2 and FGF-R1 on the protein level while the level of HSP70 remained unchanged. Coincidently, the tumor tissue was damaged by heat, resulting in large apoptotic and necrotic areas in regions with high MNP concentration. Taken together, thermoablative heating induced via magnetic methods can reduce the expression of tumor-related proteins and locally inactivate tumor tissue, leading to a prospectively reduced tumorigenicity of cancerous tissues. The presented data allow a deeper insight into the molecular mechanisms in relation to magnetic thermoablative tumor treatments with the aim of further improvements. Keywords: magnetic nanoparticles (MNP, thermoablation, in vivo, mouse model, breast cancer tumor

  5. r-1,t-2,3,c-4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene in humanurine: A potential biomarker for assessing polycyclic aromatic hydrocarbon metabolic activation

    Hecht, S.S.; Chen, M.L.; Yagi, H.; Jerina, D.M.; Carmella, S.G. [University of Minnesota, Minneapolis, MN (United States)


    Carcinogen metabolite phenotyping is proposed as a more reliable way to determinethe role of host metabolism in PAH-related cancer. Phenanthrene,is the simplest PAH with a bay region and is metabolized to diol epoxides by the same enzymes and with the same stereochemistry as the prototypic carcinogenic PAH, benzo(a)pyrene. The major end product of this metabolic activation pathway is r-1,t-2,3,c-4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene (trans, anti-PheT). We have developed a method for the analysis of trans, anti-PheT in human urine. r-1,t-2,4,c-3,-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene (trans,syn-PheT) was used as internal standard. After hydrolysis by beta-glucuronidase and sulfatase, solid phase extraction, and high-performance liquid chromatography collection, the sample was silylated and analyzed by gas chromatography-negative ion chemical ionization-mass spectrometry-selected ion monitoring. Thechromatograms were clean and trans, anti-PheT was detected in all human urine samples. Levels of trans, anti-PheT were 791 {+-} 363 pmol/mg creatinine (n = 20) in psoriasis patients treated with a PAH-containing ointment, 25.7 {+-} 16.8 pmol/mg creatinine (n = 32) in coke oven workers exposed to PAH, 4.58 {+-} 2.95 pmol/mg creatinine (n = 31) in smokers, and 1.51 {+-} 1.15 pmol/mg creatinine (n = 30) in nonsmokers. Levels of trans, anti-PheT correlated with levels of 1-hydroxypyrene in the urine of coke oven workers, smokers, and nonsmokers. Trans, anti-PheT appears to be an excellent biomarker of PAH uptake. Levels of trans, anti-PheT were 8,000-19,000 times higher than those can be detected in human urine.

  6. Determination of binding capacity and adsorption enthalpy between Human Glutamate Receptor (GluR1) peptide fragments and kynurenic acid by surface plasmon resonance experiments.

    Csapó, E; Majláth, Z; Juhász, Á; Roósz, B; Hetényi, A; Tóth, G K; Tajti, J; Vécsei, L; Dékány, I


    The interaction between kynurenic acid (KYNA) and two peptide fragments (ca. 30 residues) of Human Glutamate Receptor 201-300 (GluR1) using surface plasmon resonance (SPR) spectroscopy was investigated. Because of the medical interest in the neuroscience, GluR1 is one of the important subunits of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPAR). AMPARs are ionotoropic glutamate receptors, which are mediating fast synaptic transmission and are crucial for plasticity in the brain. On the other hand, KYNA has been suggested to have neuroprotective activity and it has been considered for apply in therapy in certain neurobiological disorders. In this article the adsorption of the GluR1201-230 and GluR1231-259 peptides were studied on gold biosensor chip. The peptides were chemically bonded onto the gold surface via thiol group of L-cysteine resulted in the formation of peptide monolayer on the SPR chip surface. Because the GluR1231-259 peptide does not contain L-cysteine the Val256 was replaced by Cys256. The cross sectional area and the surface orientation of the studied peptides were determined by SPR and theoretical calculations (LOMETS) as well. The binding capability of KYNA on the peptide monolayer was studied in the concentration range of 0.1-5.0 mM using 150 mM NaCl ionic strength at pH 7.4 (±0.02) in phosphate buffer solutions. In order to determine the binding enthalpy the experiments were carried out between +10°C and +40°C. The heat of adsorption was calculated by using adsorption isotherms at different surface loading of KYNA on the SPR chip.

  7. The neuropeptide TLQP-21 opposes obesity via C3aR1-mediated enhancement of adrenergic-induced lipolysis.

    Cero, Cheryl; Razzoli, Maria; Han, Ruijun; Sahu, Bhavani Shankar; Patricelli, Jessica; Guo, ZengKui; Zaidman, Nathan A; Miles, John M; O'Grady, Scott M; Bartolomucci, Alessandro


    Obesity is characterized by excessive fat mass and is associated with serious diseases such as type 2 diabetes. Targeting excess fat mass by sustained lipolysis has been a major challenge for anti-obesity therapies due to unwanted side effects. TLQP-21, a neuropeptide encoded by the pro-peptide VGF (non-acronymic), that binds the complement 3a receptor 1 (C3aR1) on the adipocyte membrane, is emerging as a novel modulator of adipocyte functions and a potential target for obesity-associated diseases. The molecular mechanism is still largely uncharacterized. We used a combination of pharmacological and genetic gain and loss of function approaches. 3T3-L1 and mature murine adipocytes were used for in vitro experiments. Chronic in vivo experiments were conducted on diet-induced obese wild type, β1, β2, β3-adrenergic receptor (AR) deficient and C3aR1 knockout mice. Acute in vivo lipolysis experiments were conducted on Sprague Dawley rats. We demonstrated that TLQP-21 does not possess lipolytic properties per se. Rather, it enhances β-AR activation-induced lipolysis by a mechanism requiring Ca(2+) mobilization and ERK activation of Hormone Sensitive Lipase (HSL). TLQP-21 acutely potentiated isoproterenol-induced lipolysis in vivo. Finally, chronic peripheral TLQP-21 treatment decreases body weight and fat mass in diet induced obese mice by a mechanism involving β-adrenergic and C3a receptor activation without associated adverse metabolic effects. In conclusion, our data identify an alternative pathway modulating lipolysis that could be targeted to diminish fat mass in obesity without the side effects typically observed when using potent pro-lipolytic molecules.

  8. DdrA, DdrD, and PprA: components of UV and mitomycin C resistance in Deinococcus radiodurans R1.

    Kathiresan Selvam

    Full Text Available Mutants created by deleting the ddrA, ddrB, ddrC, ddrD, and pprA loci of Deinococcus radiodurans R1alone and in all possible combinations of pairs revealed that the encoded gene products contribute to this species' resistance to UV light and/or mitomycin C. Deleting pprA from an otherwise wild type cell sensitizes the resulting strain to UV irradiation, reducing viability by as much as eight fold relative to R1. If this deletion is introduced into a ΔddrA or ΔddrD background, the resulting strains become profoundly sensitive to the lethal effects of UV light. At a fluence of 1000 Jm⁻², the ΔddrA ΔpprA and ΔddrD ΔpprA strains are 100- and 1000-fold more sensitive to UV relative to the strain that has only lost pprA. Deletion of ddrA results in a 100 fold increase in strain sensitivity to mitomycin C, but in backgrounds that combine a deletion of ddrA with deletions of either ddrC or ddrD, mitomycin resistance is restored to wild type levels. Inactivation of ddrB also increases D. radiodurans sensitivity to mitomycin, but unlike the ddrA mutant deleting ddrC or ddrD from a ΔddrB background further increases that sensitivity. Despite the effect that loss of these gene products has on DNA damage resistance, none appear to directly affect either excision repair or homologous recombination suggesting that they participate in novel processes that facilitate tolerance to UV light and interstrand crosslinks in this species.

  9. Slow feedback inhibition in the CA3 area of the rat hippocampus by synergistic synaptic activation of mGluR1 and mGluR5.

    Mori, Masahiro; Gerber, Urs


    Interneurons are critical in regulating the excitability of principal cells in neuronal circuits, thereby modulating the output of neuronal networks. We investigated synaptically evoked inhibitory responses in CA3 pyramidal cells mediated by metabotropic glutamate receptors (mGluRs) expressed somatodendritically by interneurons. Although pharmacological activation of mGluRs in interneurons has been shown to enhance their excitability, the inability to record mGluR-mediated synaptic responses has precluded detailed characterization of mGluR function in hippocampal interneurons. We found that a single extracellular pulse in CA3 stratum pyramidale was sufficient to induce disynaptic inhibitory responses mediated by postsynaptic mGluRs of the interneurons in CA3 pyramidal cells of hippocampal slice cultures. The disynaptic inhibitory response followed a short-latency monosynaptic inhibitory response, and was observed at stimulus intensities evoking half-maximal monosynaptic IPSCs. Synergistic activation of mGluR1 and mGluR5 was required to induce the full inhibitory response. When recordings were obtained from interneurons in CA3 stratum radiatum or stratum oriens, a single extracellular stimulus induced a slow inward cationic current with a time course corresponding to the slow inhibitory response measured in pyramidal cells. DCG IV, a group II mGluR agonist, which specifically blocks synaptic transmission through mossy fibres, had no effect on mGluR-mediated synaptic responses in interneurons, suggesting that feed-forward inhibition via mossy fibres is not involved. Thus, postsynaptic mGluR1 and mGluR5 in hippocampal interneurons cooperatively mediate slow feedback inhibition of CA3 pyramidal cells. This mechanism may allow interneurons to monitor activity levels from populations of neighbouring principal cells to adapt inhibitory tone to the state of the network.

  10. Validation of the betaeta-s.t.a.r. 1 + 1 for rapid screening of residues of beta-lactam antibiotics in milk.

    Reybroeck, W; Ooghe, S; De Brabander, H F; Daeseleire, E


    The 2-min protocol (1 + 1) for the betaeta-s.t.a.r. (manufactured by Neogen Corporation, Lansing, MI, USA) was validated at the Technology and Food Science Unit of the Institute for Agricultural and Fisheries Research according to Commission Decision 2002/657/EC. The test was very selective for the group of beta-lactam compounds: the only interference found was by clavulanic acid at 2500 microg kg(-1) and above. The modified protocol (betaeta-s.t.a.r. 1 + 1) detected all beta-lactams with a maximum residue limit (MRL) in milk, but not all these compounds were detected at their respective MRL. The detection of cefalexin (detection capability = 6000 microg kg(-1); MRL = 100 microg kg(-1)) and penethamate (detection capability = 80 microg kg(-1); MRL = 4 microg kg(-1)) was especially poor, and also ceftiofur was only detected from 500 microg kg(-1) (MRL = 100 microg kg(-1)). The repeatability of the reader and of the test was very good. The test was very robust: test results were not significantly influenced by small changes in the test protocol, by the milk composition or by the type of milk. The test was also suitable to test the milk of animal species other than cow. Favourable results were obtained in testing monitoring samples, in two national ring trials, and in an international proficiency test. The betaeta-s.t.a.r. 1 + 1 is a very fast, simple, and reliable test that could be used at the farm level to prevent tanker milk contamination by beta-lactams.

  11. Validation of the βeta-s.t.a.r. 1 + 1 for rapid screening of residues of β-lactam antibiotics in milk

    Reybroeck, W.; Ooghe, S.; De Brabander, H.F.; Daeseleire, E.


    The 2-min protocol (1 + 1) for the βeta-s.t.a.r. (manufactured by Neogen Corporation, Lansing, MI, USA) was validated at the Technology and Food Science Unit of the Institute for Agricultural and Fisheries Research according to Commission Decision 2002/657/EC. The test was very selective for the group of β-lactam compounds: the only interference found was by clavulanic acid at 2500 μg kg−1 and above. The modified protocol (βeta-s.t.a.r. 1 + 1) detected all β-lactams with a maximum residue limit (MRL) in milk, but not all these compounds were detected at their respective MRL. The detection of cefalexin (detection capability = 6000 μg kg−1; MRL = 100 μg kg−1) and penethamate (detection capability = 80 μg kg−1; MRL = 4 μg kg−1) was especially poor, and also ceftiofur was only detected from 500 μg kg−1 (MRL = 100 μg kg−1). The repeatability of the reader and of the test was very good. The test was very robust: test results were not significantly influenced by small changes in the test protocol, by the milk composition or by the type of milk. The test was also suitable to test the milk of animal species other than cow. Favourable results were obtained in testing monitoring samples, in two national ring trials, and in an international proficiency test. The βeta-s.t.a.r. 1 + 1 is a very fast, simple, and reliable test that could be used at the farm level to prevent tanker milk contamination by β-lactams. PMID:20512709

  12. Engineered Deinococcus radiodurans R1 with NiCoT genes for bioremoval of trace cobalt from spent decontamination solutions of nuclear power reactors.

    Gogada, Raghu; Singh, Surya Satyanarayana; Lunavat, Shanti Kumari; Pamarthi, Maruthi Mohan; Rodrigue, Agnes; Vadivelu, Balaji; Phanithi, Prakash-Babu; Gopala, Venkateswaran; Apte, Shree Kumar


    The aim of the present work was to engineer bacteria for the removal of Co in contaminated effluents. Radioactive cobalt ((60)Co) is known as a major contributor for person-sievert budgetary because of its long half-life and high γ-energy values. Some bacterial Ni/Co transporter (NiCoT) genes were described to have preferential uptake for cobalt. In this study, the NiCoT genes nxiA and nvoA from Rhodopseudomonas palustris CGA009 (RP) and Novosphingobium aromaticivorans F-199 (NA), respectively, were cloned under the control of the groESL promoter. These genes were expressed in Deinococcus radiodurans in reason of its high resistance to radiation as compared to other bacterial strains. Using qualitative real time-PCR, we showed that the expression of NiCoT-RP and NiCoT-NA is induced by cobalt and nickel. The functional expression of these genes in bioengineered D. radiodurans R1 strains resulted in >60 % removal of (60)Co (≥5.1 nM) within 90 min from simulated spent decontamination solution containing 8.5 nM of Co, even in the presence of >10 mM of Fe, Cr, and Ni. D. radiodurans R1 (DR-RP and DR-NA) showed superior survival to recombinant E. coli (ARY023) expressing NiCoT-RP and NA and efficiency in Co remediation up to 6.4 kGy. Thus, the present study reports a remarkable reduction in biomass requirements (2 kg) compared to previous studies using wild-type bacteria (50 kg) or ion-exchanger resins (8000 kg) for treatment of ~10(5)-l spent decontamination solutions (SDS).

  13. Globular adiponectin, acting via AdipoR1/APPL1, protects H9c2 cells from hypoxia/reoxygenation-induced apoptosis.

    Min Park

    Full Text Available Cardiomyocyte apoptosis is an important remodeling event contributing to heart failure and adiponectin may mediate cardioprotective effects at least in part via attenuating apoptosis. Here we used hypoxia-reoxygenation (H/R induced apoptosis in H9c2 cells to examine the effect of adiponectin and cellular mechanisms of action. We first used TUNEL labeling in combination with laser scanning cytometry to demonstrate that adiponectin prevented H/R-induced DNA fragmentation. The anti-apoptotic effect of adiponectin was also verified via attenuation of H/R-induced phosphatidylserine exposure using annexin V binding. H/R-induced apoptosis via the mitochondrial-mediated intrinsic pathway of apoptosis as assessed by cytochrome c release into cytosol and caspase-3 activation, both of which were attenuated by adiponectin. Mechanistically, we demonstrated that adiponectin enhanced anti-oxidative potential in these cells which led to attenuation of the increase in intracellular reactive oxygen species (ROS caused by H/R. To further address the mechanism of adiponctins anti-apoptotic effects we used siRNA to efficiently knockdown adiponectin receptor (AdipoR1 expression and found that this attenuated the protective effects of adiponectin on ROS production and caspase 3 activity. Knockdown of APPL1, an important intracellular binding partner for AdipoR, also significantly reduced the ability of adiponectin to prevent H/R-induced ROS generation and caspase 3 activity. In summary, H/R-induced ROS generation and activation of the intrinsic apoptotic pathway was prevented by adiponectin via AdipoR1/APPL1 signaling and increased anti-oxidant potential.

  14. A High-Resolution Foreground Model for the MWA EoR1 Field: Model and Implications for EoR Power Spectrum Analysis

    Procopio, P.; Wayth, R. B.; Line, J.; Trott, C. M.; Intema, H. T.; Mitchell, D. A.; Pindor, B.; Riding, J.; Tingay, S. J.; Bell, M. E.; Callingham, J. R.; Dwarakanath, K. S.; For, Bi-Qing; Gaensler, B. M.; Hancock, P. J.; Hindson, L.; Hurley-Walker, N.; Johnston-Hollitt, M.; Kapińska, A. D.; Lenc, E.; McKinley, B.; Morgan, J.; Offringa, A.; Staveley-Smith, L.; Wu, Chen; Zheng, Q.


    The current generation of experiments aiming to detect the neutral hydrogen signal from the Epoch of Reionisation (EoR) is likely to be limited by systematic effects associated with removing foreground sources from target fields. In this paper, we develop a model for the compact foreground sources in one of the target fields of the MWA's EoR key science experiment: the `EoR1' field. The model is based on both the MWA's GLEAM survey and GMRT 150 MHz data from the TGSS survey, the latter providing higher angular resolution and better astrometric accuracy for compact sources than is available from the MWA alone. The model contains 5 049 sources, some of which have complicated morphology in MWA data, Fornax A being the most complex. The higher resolution data show that 13% of sources that appear point-like to the MWA have complicated morphology such as double and quad structure, with a typical separation of 33 arcsec. We derive an analytic expression for the error introduced into the EoR two-dimensional power spectrum due to peeling close double sources as single point sources and show that for the measured source properties, the error in the power spectrum is confined to high k ⊥ modes that do not affect the overall result for the large-scale cosmological signal of interest. The brightest 10 mis-modelled sources in the field contribute 90% of the power bias in the data, suggesting that it is most critical to improve the models of the brightest sources. With this hybrid model, we reprocess data from the EoR1 field and show a maximum of 8% improved calibration accuracy and a factor of two reduction in residual power in k-space from peeling these sources. Implications for future EoR experiments including the SKA are discussed in relation to the improvements obtained.

  15. Determination of Notoginsenoside R1,Ginsenoside Rg1 in Xinkeshu Capsule by HPLC-ELSD%HPLC-ELSD测定心可舒胶囊中三七皂苷R1、人参皂苷Rg1的含量

    黄秋妹; 曹智启; 丁沐淦


    OBJECTIVE To determine the content of notoginsenoside R1, ginsenoside Rg1 in Xinkeshu capsule. METHODS The notoginsenoside R1, ginsenoside Rg1 in Xinkeshu capsule were separated on a COSMOSIL 5C18-PAQ Packed column(250 mm × 4.6 mm, 5μm). The mobile phase was acetonitrile-water(27 : 73) at the flow rate of 1 mL·min-1. The analytes were detected using ELSD. The drift tube temperature was 41 ℃, and gas press was 350 kPa. RESULTS The responses of notoginsenoside R1, ginsenoside Rgl were linear in the ranges of 0.123 2-6.16 ug and 0.327 2-16.36 ug, respectively. The average recoveries were 98.7% and 99.2% with RSD 2.72% and 1.54%, respectively. CONCLUSION The described method is simple and accurate. It can be used for the quality control of Xinkeshu capsule.%目的 建立心可舒胶囊中三七皂苷R1、人参皂苷Pg1的含量测定方法.方法 以COSMOSIL 5C18-PAQ Packed Column(250 mm×4.6 mm,5 μm)为色谱柱,以乙腈-水(27∶73)为流动相,流速为1 mL·min-1,ELSD为检测器,漂移管温度:41℃;雾化器(氮气)压力∶350 kPa.结果 三七皂苷R1、人参皂苷Rg1的线性范围分别为0.1232~6.16,0.3272~16.36μg,r值分别为0.9999,0.9997;回收率分别为98.7%,99.2%,RSD分别为2.72%,1.54%.结论 该方法简便,准确,灵敏度高,重复性好,可作为心可舒胶囊的含量测定方法.

  16. 十字花科黑腐病菌中转录调控因子HpaR1调控一个纤维素酶基因的表达%The Transcriptional Regulator HpaR1 of Xanthomonas campestris pv.campe-stris (Xcc) 8004 Regulates the Expression of a Extracellular Cellulase Gene

    薛番艳; 苏辉昭; 刘兴艳; 梁伟; 李磊; 李瑞芳; 陆光涛


    十字花科黑腐病菌(Xcc8004)中的一个转录调控因子XC_2736 (HpaR1)在致病过程中具有重要的作用.前期研究发现该转录调控因子可能调控胞外纤维素酶的合成.为了解HpaR1对纤维素酶的转录调控机理,本研究对HpaR1进行原核表达纯化,并与488 bp的包含XC_0639的启动子区DNA片段进行凝胶电泳迁移率试验,发现HpaR1与XC_0639启动子可以发生结合.将488 bp的XC_0639的启动子DNA片段与报告基因gus融合,构建XC_0639的报告质粒pGUS0639r,分别导入野生型8004菌株和缺失突变体DM2736中,分析发现在突变体背景下GUS的表达水平比野生型背景明显降低.表明HpaR1正调控XC_0639的表达.构建XC_0639的极性整合突变体PK0639,检测发现PK0639几乎丧失胞外纤维素酶的活力;通过功能反式互补构建的互补菌株CPK0639可以恢复纤维素酶活性.研究结果表明HpaR1通过调控纤维素酶基因XC_0639的表达来调控细胞的纤维素酶活性.本研究为更深入地了解HpaR1如何调控细菌生理生化功能奠定了基础.%The transcriptional regulator XC-2736 {HpaRl) of X. campestris pv. campestris (Xcc) 8004 plays important roles in regulating bacterial diverse biological processes. Previous work showed that this transcriptional regulator might play role on extracellular cellulase. To illustrate the regulatory mechanism, HpaRl was over expressed, and electrophoretic mobility shift assays (EMSA) were performed using HpaRl protein and the promoter fragment end XCA-p of XC_0639. Results showed that HpaRl could specifically bind to the promoter fragment of XC_0639. The reporter plasmid pGUS0639r was constructed by fusing the promoter of XC_0639 with gus gene, and them been introduced into wild type strain 8004 and the HpaRl mutant DM2736, respectively. The expression level of GUS decreased significantly in the mutant, compared to the wild type background, suggesting HpaRl positively regulates the expression of XC_0639

  17. Color regulation in the archaebacterial phototaxis receptor phoborhodopsin (sensory rhodopsin II)

    Takahashi, T.; Yan, B.; Mazur, P.; Derguini, F.; Nakanishi, K.; Spudich, J.L. (Albert Einstein College of Medicine, Bronx, NY (USA))


    Phoborhodopsin, a repellent phototaxis receptor in Halobacterium halobium, exhibits vibrational fine structure, a feature that has not been identified for any other rhodopsin pigment at physiological temperatures. This conclusion follows form analysis of the absorption properties of the pigment in H. halobium membranes containing native retinal and an array of retinal analogues. The absorption spectrum of the native pigment has a maximum at 487 nm with a pronounced shoulder at 460 nm; however, the bandwidth is that expected for a single retinylidene species. Gaussian band-shape simulation with a spacing corresponding to the vibrational frequencies of polyene stretching modes reproduces the structured absorption spectra of native pigment as well as of analogue phoborhodopsin. Absorption shifts produced by a series of dihydroretinal and other retinal analogues strongly indicate that the dominant factor regulating the color of the pigment is planarization of the retinal ring with respect to the polyene chain.

  18. Color regulation in the archaebacterial phototaxis receptor phoborhodopsin (sensory rhodopsin II).

    Takahashi, T; Yan, B; Mazur, P; Derguini, F; Nakanishi, K; Spudich, J L


    Phoborhodopsin, a repellent phototaxis receptor in Halobacterium halobium, exhibits vibrational fine structure, a feature that has not been identified for any other rhodopsin pigment at physiological temperatures. This conclusion follows form analysis of the absorption properties of the pigment in H. halobium membranes containing native retinal and an array of retinal analogues. The absorption spectrum of the native pigment has a maximum at 487 nm with a pronounced shoulder at 460 nm; however, the bandwidth is that expected for a single retinylidene species. Gaussian band-shape simulation with a spacing corresponding to the vibrational frequencies of polyene stretching modes reproduces the structured absorption spectra of native pigment as well as of analogue phoborhodopsin. Absorption shifts produced by a series of dihydroretinal and other retinal analogues strongly indicate that the dominant factor regulating the color of the pigment is planarization of the retinal ring with respect to the polyene chain.

  19. Differential Contributions of the Complement Anaphylotoxin Receptors C5aR1 and C5aR2 to the Early Innate Immune Response against Staphylococcus aureus Infection

    Sarah A. Horst


    Full Text Available The complement anaphylatoxin C5a contributes to host defense against Staphylococcus aureus. In this study, we investigated the functional role of the two known C5a receptors, C5aR1 and C5aR2, in the host response to S. aureus. We found that C5aR1−/− mice exhibited greater susceptibility to S. aureus bloodstream infection than wild type and C5aR2−/− mice, as demonstrated by the significantly higher bacterial loads in the kidneys and heart at 24 h of infection, and by the higher levels of inflammatory IL-6 in serum. Histological and immunohistochemistry investigation of infected kidneys at 24 h after bacterial inoculation revealed a discrete infiltration of neutrophils in wild type mice but already well-developed abscesses consisting of bacterial clusters surrounded by a large number of neutrophils in both C5aR1−/− and C5aR2−/− mice. Furthermore, blood neutrophils from C5aR1−/− mice were less efficient than those from wild type or C5aR2−/− mice at killing S. aureus. The requirement of C5aR1 for efficient killing of S. aureus was also demonstrated in human blood after disrupting C5a-C5aR1 signaling using specific inhibitors. These results demonstrated a role for C5aR1 in S. aureus clearance as well as a role for both C5aR1 and C5aR2 in the orchestration of the inflammatory response during infection.

  20. Mir-338-3p Mediates Tnf-A-Induced Hepatic Insulin Resistance by Targeting PP4r1 to Regulate PP4 Expression.

    Dou, Lin; Wang, Shuyue; Sun, Libo; Huang, Xiuqing; Zhang, Yang; Shen, Tao; Guo, Jun; Man, Yong; Tang, Weiqing; Li, Jian


    Insulin resistance is a critical factor contributing to the pathogenesis of type 2 diabetes and other metabolic diseases. Recent studies have indicated that miR-338-3p plays an important role in cancer. Here, we investigated whether miR-338-3p mediates tumour necrosis factor-α (TNF-α)-induced hepatic insulin resistance. The activation of the insulin signalling pathway and the level of glycogenesis were examined in the livers of the db/db and high fat diet (HFD)-fed mice and in HEP1-6 cells transfected with miR-338-3p mimic or inhibitor. Computational prediction of microRNA target, luciferase assay and Western blot were used to assess the miR-338-3p target. Chromatin immunoprecipitation (ChIP) assay was used to determine the transcriptional regulator of miR-338-3p. miR-338-3p was down-regulated in the livers of the db/db, HFD-fed and TNF-α-treated C57BL/6J mice, as well as in mouse HEP1-6 hepatocytes treated with TNF-α. Importantly the down-regulation of miR-338-3p induced insulin resistance, as indicated by impaired glucose tolerance and insulin tolerance. Further research showed that the down-regulated miR-338-3p resulted in the impaired AKT/ glycogen synthase kinase 3 beta (GSl·Gβ) signalling pathway and glycogen synthesis. In contrast, hepatic over-expression of miR-338-3p rescued the TNF-α-induced insulin resistance. Moreover, protein phosphatase 4 regulator subunit 1 (PP4R1) was identified as a direct target of miR-338-3p that mediated hepatic insulin signalling by regulating protein phosphatase 4 (PP4). Finally we identified hepatic nuclear factor 4 alpha (HNF-4α) as the transcriptional regulator of miRNA-338-3p. Our studies provide novel insight into the critical role and molecular mechanism by which miR-338-3p is involved in TNF-α-induced hepatic insulin resistance. miR-338-3p might mediate TNF-α-induced hepatic insulin resistance by targeting PP4R1 to regulate PP4 expression. © 2017 The Author(s). Published by S. Karger AG, Basel.

  1. Glucocorticoid-induced tethered transrepression requires SUMOylation of GR and formation of a SUMO-SMRT/NCoR1-HDAC3 repressing complex

    Hua, Guoqiang; Ganti, Krishna Priya; Chambon, Pierre


    Upon binding of a glucocorticoid (GC), the GC receptor (GR) can exert one of three transcriptional regulatory functions. We recently reported that SUMOylation of the GR at position K293 in humans (K310 in mice) within the N-terminal domain is indispensable for GC-induced evolutionary conserved inverted repeated negative GC response element (IR nGRE)-mediated direct transrepression. We now demonstrate that the integrity of this GR SUMOylation site is mandatory for the formation of a GR-small ubiquitin-related modifiers (SUMOs)-SMRT/NCoR1-HDAC3 repressing complex, which is indispensable for NF-κB/AP1-mediated GC-induced tethered indirect transrepression in vitro. Using GR K310R mutant mice or mice containing the N-terminal truncated GR isoform GRα-D3 lacking the K310 SUMOylation site, revealed a more severe skin inflammation than in WT mice. Importantly, cotreatment with dexamethasone (Dex) could not efficiently suppress a 12-O-tetradecanoylphorbol-13-acetate (TPA)–induced skin inflammation in these mutant mice, whereas it was clearly decreased in WT mice. In addition, in mice selectively ablated in skin keratinocytes for either nuclear receptor corepressor 1 (NCoR1)/silencing mediator for retinoid or thyroid-hormone receptors (SMRT) corepressors or histone deacetylase 3 (HDAC3), Dex-induced tethered transrepression and the formation of a repressing complex on DNA-bound NF-κB/AP1 were impaired. We previously suggested that GR ligands that would lack both (+)GRE-mediated transactivation and IR nGRE-mediated direct transrepression activities of GCs may preferentially exert the therapeutically beneficial GC antiinflammatory properties. Interestingly, we now identified a nonsteroidal antiinflammatory selective GR agonist (SEGRA) that selectively lacks both Dex-induced (+)GRE-mediated transactivation and IR nGRE-mediated direct transrepression functions, while still exerting a tethered indirect transrepression activity and could therefore be clinically lesser

  2. Antidepressant-like effects of mGluR1 and mGluR5 antagonists in the rat forced swim and the mouse tail suspension tests.

    Belozertseva, I V; Kos, T; Popik, P; Danysz, W; Bespalov, A Y


    Drugs that act to reduce glutamatergic neurotransmission such as NMDA receptor antagonists exert antidepressant-like effects in a variety of experimental paradigms, but their therapeutic application is limited by undesired side effects. In contrast, agents that reduce glutamatergic tone by blocking type I metabotropic glutamate receptors have been suggested to have more a favorable side-effect profile. The present study aimed to compare the effects of mGluR1 antagonist (EMQMCM; JNJ16567083, 3-ethyl-2-methyl-quinolin-6-yl)-(4-methoxy-cyclohexyl)-methanone methanesulfonate, 0.156-10 mg/kg) and mGluR5 antagonist (MTEP, [(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine, 1.25-10 mg/kg) in two behavioral screening assays commonly used to assess antidepressant-like activity. In the modified forced swim test in rats, imipramine (used as a positive control) decreased immobility (MED 40 mg/kg) and increased the duration of escape-oriented (climbing and diving; MED 20 mg/kg) behaviors. Both EMQMCM and MTEP decreased the floating duration (MED 1.25 and 2.5 mg/kg) and increased the duration of mobile behaviors (paddling and swimming; MED 2.5 and 5 mg/kg). EMQMCM but not MTEP increased the duration of escape behaviors (climbing and diving; MED 1.25 mg/kg). In the mouse tail suspension test, EMQMCM (5 but not 2.5, 10 and 25 mg/kg), 2-methyl-6-(phenylethynyl)-pyridine (MPEP, 10 but not 1 mg/kg) and MTEP (MED 25 mg/kg) decreased immobility scores. For EMQMCM, the dose-effect relationship was biphasic. With the exception of EMQMCM (10 mg/kg), locomotor activity in mice was not affected by treatments. The present study therefore suggests that acute blockade of mGluR5 and also of mGluR1 exerts antidepressant-like effects in behavioral despair tests in rats and mice.

  3. Impact of uranium concentration reduction in side plates of the fuel elements of IEA-R1 reactor on neutronic and thermal hydraulic analyses; Impacto da reducao na concentracao de uranio nas placas laterais dos elementos combustiveis do reator IEA-R1 nas analises neutronica e termo-hidraulica

    Rios, Ilka Antonia


    This master thesis presents a study to verify the impact of the uranium concentration reduction in the side plates of the reactor IEA-R1 fuel elements on the neutronic and thermal-hydraulic analyses. To develop such study, a previous IPEN-CNEN/SP research was reproduced by simulating the fuel elements burn-up, with side plate uranium density reduced to 50, 60 and 70% of the standard fuel element plates. This research begins with the neutronic analysis using the computer code HAMMER and the first step consists in the calculation of the cross section of all materials presented at the reactor core, with their initial concentration; the second step consists in the calculation of the fast and thermal neutron group fluxes and power densities for fuel elements using the computer code CITATION. HAMMER output data is used as input data. Once the neutronic analysis is finished and the most critical fuel elements with highest power density have been defined, the thermal-hydraulics analysis begins. This analysis uses MCTR-IEA-R1 thermal-hydraulics model, which equations are solved by commercial code EES. Thermalhydraulics analysis input is the power density data calculated by CITATION: it is considered the highest power density on each fuel element, where there is a higher energy release and, consequently, higher temperatures. This data is used on energy balance equations to calculate temperatures on critical fuel element regions. Reactor operation comparison for three different uranium densities on fuel side plates is presented. Uranium density reduction contributes to the cladding surface temperature to remain below the established limit, as reactor operation safety requirement and it does not affect significantly fuel element final burn-up nor reactor reactivity. The reduction of uranium in the side plates of the fuel elements of the IEA-R1