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Selected AbstractsCassette mutagenesis of lysine 130 of human glutamate dehydrogenaseFEBS JOURNAL, Issue 11 2001An essential residue in catalysis It has been suggested that reactive lysine residue(s) may play an important role in the catalytic activities of glutamate dehydrogenase (GDH). There are, however, conflicting views as to whether the lysine residues are involved in Schiff's base formation with catalytic intermediates, stabilization of negatively charged groups or the carbonyl group of 2-oxoglutarate during catalysis, or some other function. We have expanded on these speculations by constructing a series of cassette mutations at Lys130, a residue that has been speculated to be responsible for the activity of GDH and the inactivation of GDH by pyridoxal 5,-phosphate (PLP). For these studies, a 1557-bp gene that encodes human GDH has been synthesized and inserted into Escherichia coli expression vectors. The mutant enzymes containing Glu, Gly, Met, Ser, or Tyr at position 130, as well as the wild-type human GDH encoded by the synthetic gene, were efficiently expressed as a soluble protein and are indistinguishable from that isolated from human and bovine tissues. Despite an approximately 400-fold decrease in the respective apparent Vmax of the Lys130 mutant enzymes, apparent Km values for NADH and 2-oxoglutarate were almost unchanged, suggesting the direct involvement of Lys130 in catalysis rather than in the binding of coenzyme or substrate. Unlike the wild-type GDH, the mutant enzymes were unable to interact with PLP, indicating that Lys130 plays an important role in PLP binding. The results with analogs of PLP suggest that the aldehyde moiety of PLP, but not the phosphate moiety, is required for efficient binding to GDH. [source] Changes in expression and activity levels of ecto-5,-nucleotidase/CD73 along the mouse female estrous cycleACTA PHYSIOLOGICA, Issue 2 2010E. Aliagas Abstract Aim:, Extracellular ATP and its hydrolysis product adenosine modulate various reproductive functions such as those requiring contraction, hormone synthesis and maintenance of fluid composition. Moreover, adenosine is a key molecule for sperm capacitation. Extracellular nucleotide and nucleoside levels are affected by cell surface ectonucleotidases, amongst which the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) family is the most abundant and effective to hydrolyse ATP and ADP to AMP. In the female reproductive tract three members of this family have been recently identified: NTPDase1, NTPDase2 and NTPDase3 (Histochem. Cell Biol.131, 2009, 615). The purpose of the present study was to characterize in this system the expression profile of ecto-5,-nucleotidase (CD73), the enzyme generating adenosine from AMP. Methods:, Immunological techniques and in situ enzymatic assays were used to characterize the ecto-5,-nucleotidase expression in the mouse female reproductive tract along the four stages of the estrous cycle, that were determined by vaginal smear examination. Results:, Ecto-5,-nucleotidase was abundantly detected in the corpora lutea of the ovaries, as well as in several epithelia, such as that of oviducts, uterus and endometrial glands. Marked changes in endometrial ecto-5,-nucleotidase expression and activity along the estrous cycle are described, these being maximum at estrus phase, coinciding with optimal female sexual receptivity. Conclusion:, The adenosine generated thereby, besides other functions, might contribute to sperm capacitation, thus significantly influencing fertility. [source] Pituitary and autonomic responses to cold exposures in manACTA PHYSIOLOGICA, Issue 4 2005J. Leppäluoto Abstract This review presents hormonal responses to various cold exposures and their calorigenic effects in man and some animals. Previous studies in rats have shown that cold exposures activate the hypothalamic-pituitary-thyroid axis. Increased thyroid hormone concentrations lead to heat production via general stimulation of metabolism (obligatory thermogenesis) and possibly via activation of thyroid hormone receptors and uncoupling protein 1 (UCP 1) and deiodinase enzyme genes in the brown adipose tissue (BAT). In human subjects long-term cold exposures do not seem to activate the pituitary-thyroid axis, but rather accelerate the elimination of triiodothyronine (T3), leading to low serum concentrations of free T3 hormone. In corollary to this a hypothyreotic condition with increased serum thyroid-stimulating hormone and impaired mood and cognitive performance can be observed after long-term cold exposures such as wintering. During cold exposures the sympathetic nerve system is activated and noradrenaline is released to blood circulation and to BAT, where it leads to production of cAMP, lipolysis and free fatty acids. Free fatty acids open the mitochondrial proton channel protein in BAT. Protons enter the mitochondria and inhibit ATP synthesis (uncoupling). By this way energy is transformed into heat (facultatory or adaptive thermogenesis). In adult human subjects the amount of BAT is small and adaptive thermogenesis (non-shivering thermogenesis) has a smaller role. UCP 1 with other uncoupling proteins may have other functions in the control of body weight, sugar balance and formation of reactive oxygen species. [source] Innovation,What's Design Got to Do with It?DESIGN MANAGEMENT REVIEW, Issue 1 2004Bettina von Stamm If the goal is innovation, then Bettina von Stamm is sure design is an essential component in realizing that goal. She celebrates the differences between designers and other functions and documents how their unique perspectives can improve business performance, help expand market share, and leverage bottom-line results. She believes design must be an in-house expertise and outlines strategies to integrate design and designers into the innovation effort. [source] Managing Design for Competitive Advantage A Process ApproachDESIGN MANAGEMENT REVIEW, Issue 4 2000Eric M. Olson FOR EXECUTIVES who want to strengthen the design/ business relationship, Eric Olson, Stanley Slater, and Rachel Cooper make some clear recommendations. Make sure designers and design managers understand the organization's competitive strategy; enumerate the design elements inherent in that strategy; nurture open communication between design and other functions; develop design briefs that stimulate creativity at the same time they reinforce business strategy; and measure performance. It's advice that's both sound and challenging. [source] Differential expression of PKC beta II in the rat organ of CortiEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 10 2007S. Ladrech Abstract To investigate a possible involvement of protein kinase C (PKC) in cochlear efferent neurotransmission, we studied the expression of the calcium-dependent PKC beta II isoform in the rat organ of Corti at different postnatal ages using immunofluorescence and immunoelectron microscopy. We found evidence of PKC beta II as early as postnatal day (PND) 5 in efferent axons running in the inner spiral bundle and in Hensen cells. At PND 8, we also found PKC beta II in efferents targeting outer hair cells (OHCs), and a slight detection at the synaptic pole in the first row of the basal and middle cochlear turns. At PND 12, PKC beta II expression declined in the efferent fibres contacting OHCs, whereas expression was concentrated at the postsynaptic membrane, from the basal and middle turns. The adult-like pattern of PKC beta II distribution was observed at PND 20. Throughout the cochlea, we found PKC beta II expression in the Hensen cells, non-sensory cells involved in potassium re-cycling, and lateral efferent terminals of the inner spiral bundle. In addition, we observed expression in OHCs at the postsynaptic membrane facing the endings of the medial efferent system, with the exception of some OHCs located in the most apical region of the cochlea. These data therefore suggest an involvement of PKC beta II in both cochlear efferent neurotransmission and ion homeostasis. Among other functions, PKC beta II could play a role in the efferent control of OHC activity. [source] Covariance of tricarboxylate carrier activity and lipogenesis in liver of polyunsaturated fatty acid (n-6) fed ratsFEBS JOURNAL, Issue 22 2001Vincenzo Zara The mitochondrial tricarboxylate (citrate) carrier plays an important role in hepatic intermediary metabolism because, among other functions, it supplies the cytosol with acetyl units for fatty-acid synthesis. In this study, the effect of polyunsaturated fatty acids (PUFA, n-6) on the function of this mitochondrial transporter and on lipogenic enzyme activities was investigated by feeding rats for 4 weeks with a 15%-fat diet composed of high linoleic safflower oil. Citrate transport was strongly reduced in liver mitochondria isolated from PUFA-treated rats. A reduced transport activity was also observed when solubilized mitochondrial citrate carrier from PUFA-treated rats was reconstituted into liposomes. In the same animals, a decrease of cytosolic lipogenic enzyme activities was observed. These results indicate a coordinated modulation of citrate carrier and of lipogenic enzyme activities by PUFA feeding. Kinetic analysis of the carrier activity showed that only Vmax decreased, whereas Km was almost virtually unaffected. The PUFA-mediated effect is most likely due to the reduced mRNA level and lower content of the citrate carrier protein observed in the safflower oil-fed rats. [source] Fat stores in birds: an overlooked sink for carotenoid pigments?FUNCTIONAL ECOLOGY, Issue 3 2001J. J. Negro Summary 1,Carotenoids are responsible for the most striking colours in birds, but also play an important role as enhancers of the immune system. Consequently, a trade-off between the ornamental and health functions of carotenoids in birds has been proposed. 2,Although it is well known that birds can store carotenoids in different organs and tissues, including the fat stores, until now all field studies of the regulation of carotenoid stores have focused on plasma carotenoids. 3,Carotenoids in the fat of 44 wild Greylag Geese (Anser anser L.) wintering in south-western Spain were identified and quantified. In addition, the relationships between carotenoids and the size of the fat stores, as well as the sex and age of the geese, were analysed. 4,The major carotenoid in goose fat was lutein. This and related carotenoids are also the most prevalent pigments in bird plasma and secondary sexual characters. We also detected ,-cryptoxanthin, ,-carotene, neochrome and neoxanthin. Total carotenoid concentration was negatively correlated with the size of the fat stores. Males had higher concentrations of carotenoids than females. 5A possible explanation for these patterns is that male birds tend to have higher plasma carotenoids than females, a difference that could be transposed to fat if carotenoids diffused passively from the blood into adipose tissues. Carotenoids, however, may tend to remain in the fat stores. If this were true, fat would not be a reservoir of carotenoids, but a sink where a significant amount of these pigments would be sequestered, being no longer available for other functions. [source] Mrp2 modulates the activity of chloride channels in isolated hepatocytesHEPATOLOGY, Issue 1 2002Xinhua Li Adenosine triphosphate binding cassette family transport proteins are important organic ion transporters in hepatocytes but these molecules may also exhibit other functions. In the present study we have measured the effects of substrates of the canalicular organic ion transporter multidrug resistance associated protein 2 (Mrp2) on chloride channel activation and cell volume regulation. We found that substrates such as leukotriene D4, 17-,-estradiol glucuronide, and the leukotriene inhibitor MK-571 accelerated the activation of chloride channels by cell swelling and activated chloride channels in cytokine-pretreated hepatocytes. Two conjugated estrogens that are not Mrp2 substrates did not produce this effect. Hepatocytes derived from a strain of transport-deficient rats (TR,), which lack Mrp2 expression, showed none of these substrate effects. Coincident with their ability to activate channels, the Mrp2 substrates increased the rate of volume regulatory decrease by approximately 50% (P < .01), confirming that enhanced channel activation under this condition stimulated volume regulation. In TR-hepatocytes the Mrp2 substrate had no effect on volume regulation. In conclusion, Mrp2 plays a role in regulation of chloride channel function by reducing the lag time necessary for channel activation and consequently accelerating the process of cell volume regulation. Substrates of Mrp2 affect the ability of the protein to interact with chloride channels. These findings represent an alternative function of Mrp2 in hepatocytes. [source] Use of TLC-FID and GC-MS/FID to examine the effects of migratory state, diet and captivity on preen wax composition in White-throated Sparrows Zonotrichia albicollisIBIS, Issue 4 2010RAYMOND H. THOMAS Preen wax is important for plumage maintenance and other functions. Its chemical composition is complex, and separating and quantifying its components, commonly by gas chromatography (GC), can be challenging. We present a simple analytical system consisting of thin-layer chromatography/flame ionization detection (TLC-FID) using a solvent system of 100% toluene to analyse the complex compound classes present in preen wax. We used GC and TLC-FID to investigate the effects of migratory status, diet and captivity on the preen wax composition of White-throated Sparrows Zonotrichia albicollis, and to measure the quantity of preen wax on the head, primary and tail feathers. White-throated Sparrows produced preen wax containing only monoesters regardless of migratory state. The monoesters contained several isomers consisting of homologous series of fatty alcohols (C10,C20) and fatty acids (C13,C19) esterified together in different combinations to form monoesters with total carbon numbers ranging from C23 to C38. Weighted average monoester carbon number was greater in captive birds than in wild birds and was greater in captives fed a formulated diet enriched with sesame oil than in birds fed the same diet enriched with fish oil. Captivity and migratory state also affected the complexity of the mixture of monoesters. There was significantly more preen wax on head feathers compared with primary and tail feathers. We suggest that among its many functions, preen wax may play a role in drag reduction by affecting the physical properties of feathers, and/or the fluid flow at their surfaces. [source] Spatial distribution and differential expression of the PBAN receptor in tissues of adult Helicoverpa spp. (Lepidoptera: Noctuidae)INSECT MOLECULAR BIOLOGY, Issue 3 2007A. Rafaeli Abstract Pheromone-biosynthesis-activating neuropeptide (PBAN) regulates sex pheromone production in many female moths. PBAN-like peptides, with common FXPRLamide C-terminals are found in other insect groups where they have other functions. The ubiquity and multifunctional nature of the pyrokinin/PBAN family of peptides suggests that the PBAN receptor proteins could also be present in a variety of insect tissues with alternative functions from that of sex pheromone biosynthesis. Previously we showed the presence of the PBAN-R in Helicoverpa armigera at the protein level. In the present study we confirm the similarities between the two Helicoverpa species: armigera and zea by (1) demonstrating the presence of the receptor protein in Sf9 cells, cloned to express the HezPBAN receptor, as compared with the endogenous receptor protein, previously shown in H. armigera pheromone glands, and (2) by identifying the nucleotide sequence of the PBAN-R from mRNA of H. armigera pheromone glands. Sequences of the two Helicoverpa spp. are 98% identical with most changes taking place in the 3,-end. We demonstrate the spatial distribution of the PBAN receptor protein in membranes of H. armigera brain (Br), thoracic ganglion (TG) and ventral nerve cord (VNC). We also demonstrate the presence and differential expression of the PBAN receptor gene (using reverse transcription,polymerase chain reaction and reverse transcription,quantitative real-time polymerase chain reaction, respectively) in the neural tissues (Br, TG and VNC) of adult H. armigera female moths as compared with its presence in pheromone glands. Surprisingly, the gene for the PBAN receptor is also detected in the male tissue homologous to the female pheromone gland, the aedeagus, although the protein is undetectable and PBAN does not induce physiological (pheromone production) or cellular (cyclic-adenosine monophosphate production) responses in this tissue. Our findings indicate that PBAN or PBAN-like receptors are present in the neural tissues and may represent a neurotransmitter-like function for PBAN-like peptides. In addition, the surprising discovery of the presence of the gene encoding the PBAN receptor in the male homologous tissue, but its absence at the protein level, launches opportunities for studying molecular regulation pathways and the evolution of these G protein coupled receptors (GPCRs). [source] Russia's non-strategic nuclear forcesINTERNATIONAL AFFAIRS, Issue 3 2001David S. Yost Several factors explain the high level of support for non-strategic nuclear forces(NSNF) in Russia and the correspondingly limited interest in NSNF arms control. These include Russia's conventional military weakness, NATO's conventional military superiority, political assessments that portray NATO as threatening to Russia, and the several important functions assigned to Russia's nuclear weapons and to NSNF in particular by Russian military doctrine and policy. The Russians have made it clear that they attach great importance to NSNF in a number of ways: in their preoccupations during the NATO-Russia Founding Act negotiations in 1996-7; in their recent military exercises; in their decisions regarding NSNF modernization; in their lack of transparency in implementing their 1991-2 commitments to reduce and eliminate certain types of NSNF; and in their discussions about possibly abandoning certain nuclear arms control commitments. Russian interests in using NSNF to deter powers other than NATO (such as China), to substitute for advanced non-nuclear precision-strike systems, and to ,de-escalate' regional conflicts (among other functions attributed to NSNF) would not be modified by the course of action some observers have advocated,a unilateral withdrawal of US NSNF from Europe. Such a withdrawal would, however, damage the Western alliance's security interests. NATO has adopted the most practical objective currently available: pursuing greater transparency regarding NSNF in the NATO-Russia Permanent Joint Council. [source] Allosteric properties of hemoglobin and the plasma membrane of the erythrocyte: New insights in gas transport and metabolic modulationIUBMB LIFE, Issue 2 2008Maria Cristina De Rosa Abstract Within the red blood cell the hemoglobin molecule is subjected to modulation mechanisms, namely homo- and heterotropic interactions, which optimize its functional behavior to the specific physiological requirements. At the cellular level, these modulation mechanisms are utilized to perform a number of other functions that are not minor with respect to the basic function of oxygen transport. Here we report some key examples concerning: (i) the interaction of hemoglobin with band 3 and its influence on glucose metabolism; (ii) the role of the ligand-linked quaternary transition of hemoglobin in the control of "NO bioactivity" and of gas diffusion; (iii) the interaction of plasma membrane with the various oxidative derivatives of the hemoglobin molecule. © 2008 IUBMB IUBMB Life, 60(2): 87,93, 2008 [source] A new fate for old cells: brush cells and related elementsJOURNAL OF ANATOMY, Issue 4 2005A. Sbarbati Abstract Over the past 50 years, hundreds of studies have described those cells that are characterized by a brush of rigid apical microvilli with long rootlets, and which are found in the digestive and respiratory apparatuses. These cells have been given names such as brush cells, tuft cells, fibrillovesicular cells, multivesicular cells and caveolated cells. More recently, it has been realized that all these elements may represent a single cell type, probably with a chemosensory role, even if other functions (e.g. secretory or absorptive) seem to be possible. Very recent developments have permitted a partial definition of the chemical code characterizing these elements, revealing the presence of molecules involved in chemoreceptorial cell signalling. A molecular cascade, similar to those characterizing the gustatory epithelium, seems to be present in these elements. These new data suggest that these elements can be considered solitary chemosensory cells with the presence of the apical ,brush' as an inconsistent feature. They seem to comprise a diffuse chemosensory system that covers large areas (probably the whole digestive and respiratory apparatuses) with analogies to chemosensory systems described in aquatic vertebrates. [source] Carotenoid coloration in great black-backed gull Larus marinus reflects individual qualityJOURNAL OF AVIAN BIOLOGY, Issue 1 2006Kai O. Kristiansen Carotenoids are a large group of biochemicals, with similar properties, synthesised by bacteria, fungi, algae and plants. Vertebrates obtain these biologically active pigments through the diet, and they are a disproportionately common component of animal colour signals and play important roles in immune functions and as antioxidants. Carotenoids are believed to be a limited resource and because of the trade-off between allocation of carotenoids to signals and to other functions, carotenoid based signals are often thought to be handicap signals. The purpose of this study was to investigate the signalling potential of carotenoid-based tissue coloration in the great black-backed gull Larus marinus. The intensity of carotenoid-based coloration in bill, gape and eye-ring coloration was investigated in relation to body condition, reproductive parameters, levels of immune activity, and sexual dimorphism. In males there was a positive relationship between colour intensity and body condition, but in females no such relationship was found. However, females with high colour intensity had larger eggs and clutches. Additionally, females with high red scores tended to have high density of circulating lymphocytes. There was no sexual dimorphism in coloration and there was a negative relationship between colour intensity and sampling time, which indicates that this coloration is most intensely expressed early in the breeding season. The results in this study suggest that carotenoid-based coloration in great black-backed gull are partly condition dependent and reveal information about individual quality in both males an females. Hence, it might have evolved as an important signal for assessing the quality of potential mates. [source] Proteomics analysis of liver samples from puffer fish Takifugu rubripes exposed to excessive fluoride: An insight into molecular response to fluorosisJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 1 2010Jian Lu Abstract Comparative proteomics was performed to identify proteins in the liver of Takifugu rubripes in response to excessive fluoride exposure. Sixteen fish were randomly divided into a control group and an experimental group. The control group was raised in soft water alone (F, = 0.4 mg/L), and the experimental group was raised in the same water with sodium fluoride at a high concentration of 35 mg/L. After 3 days, proteins were extracted from the fish livers and then subjected to two-dimensional polyacrylamide gel electrophoresis analysis. The matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was applied to identify the proteins that were differentially expressed from the two groups of fish. Among an average of 816 and 918 proteins detected in the control and treated groups, respectively, 16 proteins were upregulated and 35 were downregulated (P < 0.01) in the fluoride-treated group as compared with those in the control group. Twenty-four highly differentially expressed proteins were further analyzed by MALDI-TOF/TOF-MS, and eight were identified by Mascot. These eight proteins include disulfide isomerase ER-60, 4SNc-Tudor domain protein, SMC3 protein, Cyclin D1, and mitogen-activated protein kinase 10, as well as three unknown proteins. Consistent with their previously known functions, these identified proteins seem to be involved in apoptosis and other functions associated with fluorosis. These results will greatly contribute to our understanding of the effects of fluoride exposure on the physiological and biochemical functions of Takifugu and the toxicological mechanism of fluoride causing fluorosis in both fish and human. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:21,28, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20308 [source] Integrative nuclear FGFR1 signaling (INFS) as a part of a universal "feed-forward-and-gate" signaling module that controls cell growth and differentiationJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 4 2003Michal K. Stachowiak Abstract A novel signaling mechanism is described through which extracellular signals and intracellular signaling pathways regulate proliferation, growth, differentiation, and other functions of cells in the nervous system. Upon cell stimulation, fibroblast growth factor receptor-1 (FGFR1), a typically plasma membrane-associated protein, is released from ER membranes into the cytosol and translocates to the cell nucleus by an importin-,-mediated transport pathway along with its ligand, FGF-2. The nuclear accumulation of FGFR1 is activated by changes in cell contacts and by stimulation of cells with growth factors, neurotransmitters and hormones as well as by a variety of different second messengers and thus was named integrative nuclear FGFR1 signaling (INFS). In the nucleus, FGFR1 localizes specifically within nuclear matrix-attached speckle-domains, which are known to be sites for RNA Pol II-mediated transcription and co-transcriptional pre-mRNA processing. In these domains, nuclear FGFR1 colocalizes with RNA transcription sites, splicing factors, modified histones, phosphorylated RNA Pol II, and signaling kinases. Within the nucleus, FGFR1 serves as a general transcriptional regulator, as indicated by its association with the majority of active nuclear centers of RNA synthesis and processing, by the ability of nuclear FGFR1 to activate structurally distinct genes located on different chromosomes and by its stimulation of multi-gene programs for cell growth and differentiation. We propose that FGFR1 is part of a universal "feed-forward-and-gate" signaling module in which classical signaling cascades initiated by specific membrane receptors transmit signals to sequence specific transcription factors (ssTFs), while INFS elicited by the same stimuli feeds the signal forward to the common coactivator, CREB-binding protein (CBP). Activation of CBP by INFS, along with the activation of ssTFs by classical signaling cascades brings about coordinated responses from structurally different genes located at different genomic loci. © 2003 Wiley-Liss, Inc. [source] Differentiation dependent expression of TRPA1 and TRPM8 channels in IMR-32 human neuroblastoma cellsJOURNAL OF CELLULAR PHYSIOLOGY, Issue 1 2009Lauri M. Louhivuori TRPA1 and TRPM8 are transient receptor potential (TRP) channels involved in sensory perception. TRPA1 is a non-selective calcium permeable channel activated by irritants and proalgesic agents. TRPM8 reacts to chemical cooling agents such as menthol. The human neuroblastoma cell line IMR-32 undergoes a remarkable differentiation in response to treatment with 5-bromo-2-deoxyuridine. The cells acquire a neuronal morphology with increased expression of N-type voltage gated calcium channels and neurotransmitters. Here we show using RT-PCR, that mRNA for TRPA1 and TRPM8 are strongly upregulated in differentiating IMR-32 cells. Using whole cell patch clamp recordings, we demonstrate that activators of these channels, wasabi, allyl-isothiocyanate (AITC) and menthol activate membrane currents in differentiated cells. Calcium imaging experiments demonstrated that AITC mediated elevation of intracellular calcium levels were attenuated by ruthenium red, spermine, and HC-030031 as well as by siRNA directed against the channel. This indicates that the detected mRNA level correlate with the presence of functional channels of both types in the membrane of differentiated cells. Although the differentiated IMR-32 cells responded to cooling many of the cells showing this response did not respond to TRPA1/TRPM8 channel activators (60% and 90% for AITC and menthol respectively). Conversely many of the cells responding to these activators did not respond to cooling (30%). This suggests that these channels have also other functions than cold perception in these cells. Furthermore, our results suggest that IMR-32 cells have sensory characteristics and can be used to study native TRPA1 and TRPM8 channel function as well as developmental expression. J. Cell. Physiol. 221: 67,74, 2009. © 2009 Wiley-Liss, Inc [source] Condition-adapted stress and longevity gene regulation by Caenorhabditis elegans SKN-1/NrfAGING CELL, Issue 5 2009Riva P. Oliveira Summary Studies in model organisms have identified regulatory processes that profoundly influence aging, many of which modulate resistance against environmental or metabolic stresses. In Caenorhabditis elegans, the transcription regulator SKN-1 is important for oxidative stress resistance and acts in multiple longevity pathways. SKN-1 is the ortholog of mammalian Nrf proteins, which induce Phase 2 detoxification genes in response to stress. Phase 2 enzymes defend against oxygen radicals and conjugate electrophiles that are produced by Phase 1 detoxification enzymes, which metabolize lipophilic compounds. Here, we have used expression profiling to identify genes and processes that are regulated by SKN-1 under normal and stress,response conditions. Under nonstressed conditions SKN-1 upregulates numerous genes involved in detoxification, cellular repair, and other functions, and downregulates a set of genes that reduce stress resistance and lifespan. Many of these genes appear to be direct SKN-1 targets, based upon presence of predicted SKN-binding sites in their promoters. The metalloid sodium arsenite induces skn-1- dependent activation of certain detoxification gene groups, including some that were not SKN-1-upregulated under normal conditions. An organic peroxide also triggers induction of a discrete Phase 2 gene set, but additionally stimulates a broad SKN-1-independent response. We conclude that under normal conditions SKN-1 has a wide range of functions in detoxification and other processes, including modulating mechanisms that reduce lifespan. In response to stress, SKN-1 and other regulators tailor transcription programs to meet the challenge at hand. Our findings reveal striking complexity in SKN-1 functions and the regulation of systemic detoxification defenses. [source] The effects of carotenoid and food intake on caudal fin regeneration in male guppiesJOURNAL OF FISH BIOLOGY, Issue 4 2006G. R. Kolluru The trade-offs involved in allocating carotenoid pigments and food to healing and regrowing damaged caudal fin tissue v. other functions were examined in guppies Poecilia reticulata, a species in which females prefer males that display larger amounts of carotenoids in their skin. The guppies were derived from four natural populations in Trinidad that differed in resource availability but not predation intensity. Carotenoids, food and site of origin did not affect either absolute or relative fin regrowth, which suggested that fin regeneration in guppies was not constrained by carotenoid availability. It is possible that carotenoid intake influences fin regeneration in the presence of natural stressors such as predators. There was a significant negative interaction between food level in the laboratory and resource availability in the field: males from low-resource-availability sites regrew more fin tissue when raised on the high food level, and males from high-resource-availability sites regrew more fin tissue when raised on the low food level. The direction of this interaction runs counter to theoretical expectations. [source] Green Energy or Organic Food?: A Life-Cycle Assessment Comparing Two Uses of Set-Aside LandJOURNAL OF INDUSTRIAL ECOLOGY, Issue 3 2001Richard van den Broek Summary Bioenergy has a large worldwide potential in future climate change abatement, although its application may become limited by demands for land for other functions. The aim of this study was to make an environmental assessment of the use of energy crops in the Netherlands in a context that incorporates scarcity of land. A base case system was defined, consisting of conventional winter wheat production, set-aside land (1 hectare, together), and the production of coal-based electricity. Using life-cycle assessment, we compared this system with (1) a green energy system in which willow is cultivated on the set-aside land to replace the coal-based electricity and (2) an organic agriculture system in which the full hectare produces wheat under the Dutch EKO organic agriculture standard. In this way, the functional unit and the amount of land used is the same in each system. The final system comparison was based on normalized scores per environmental theme. The green energy system scored the best with respect to acidification, climate change, and energy carrier depletion. The organic food system scored best on terrestrial eco-toxicity and slightly better on the mutually related themes of seawater and seawater sediment eco-toxicity. The base case system performed slightly better with regard to eutrophication. Preferences, from an environmental point of view, for one of the systems should be determined by environmental policy priorities and the severity of local environmental problems. The case studied here shows that when climate change, energy carrier depletion, and acidification are the main drivers behind environmental policy, one should focus not on the extensification of agriculture, but rather dedicate more land to energy crops. Extensification of agriculture would be the preferred system when toxicity from pesticides is considered the main problem. [source] Heat Shock Transcription Factors and the hsp70 Induction Response in Brain and Kidney of the Hyperthermic Rat During Postnatal DevelopmentJOURNAL OF NEUROCHEMISTRY, Issue 1 2000Andrew J. Morrison Abstract : Heat shock transcription factor (HSF) 1 levels increase in brain regions and decline in kidney during postnatal rat development. In both neonatal and adult rats, levels of HSF1 protein in brain and kidney are proportional to the levels of HSF DNA-binding activity and the magnitude of heat shock protein hsp70 induction after thermal stress. There appears to be more HSF1 protein in adult brain than is needed for induction of hsp70 after thermal stress, suggesting that HSF1 may have other functions in addition to its role as a stress-inducible activator of heat shock genes. HSF2 protein levels decline during postnatal rat development in brain regions and kidney. Gel mobility shift analysis shows that HSF2 is not in a DNA-binding form in the neonatal brain and kidney, suggesting that HSF2 may not be involved in the constitutive expression of hsps in early postnatal development. There is no apparent relationship between levels of HSF2 protein and basal levels of hsp90, hsp70, heat shock cognate protein hsc70, and hsp60. [source] TorsinA in PC12 cells: Localization in the endoplasmic reticulum and response to stressJOURNAL OF NEUROSCIENCE RESEARCH, Issue 2 2003Jeffrey Hewett Abstract Most cases of early-onset torsion dystonia are caused by deletion of GAG in the coding region of the DYT1 gene encoding torsinA. This autosomal dominant neurologic disorder is characterized by abnormal movements, believed to originate from neuronal dysfunction in the basal ganglia of the human brain. The torsins (torsinA and torsinB) are members of the "ATPases associated with a variety of cellular activities" (AAA+) superfamily of proteins that mediate chaperone and other functions involved in conformational modeling of proteins, protection from stress, and targeting of proteins to cellular organelles. In this study, the intracellular localization and levels of endogenous torsin were evaluated in rat pheochromocytoma PC12 cells following differentiation and stress. TorsinA, apparent MW 37 kDa, cofractionates with markers for the microsomal/endoplasmic reticulum (ER) compartment and appears to reside primarily within the ER lumen based on protease resistance. TorsinA immunoreactivity colocalizes with the lumenal ER protein protein disulfide isomerase (PDI) and extends throughout neurites. Levels of torsinA did not increase notably in response to nerve growth factor-induced differentiation. None of the stress conditions tested, including heat shock and the unfolded protein response, affected torsinA, except for oxidative stress, which resulted in an increase in the apparent MW of torsinA and redistribution to protrusions from the cell surface. These findings are consistent with a relatively rapid covalent modification of torsinA in response to oxidative stress causing a change in state. Mutant torsinA may interfere with and/or compromise ER functions, especially in dopaminergic neurons, which have high levels of torsinA and are intrinsically vulnerable to oxidative stress. © 2003 Wiley-Liss, Inc. [source] PHOTOSYNTHETIC PERFORMANCE, LIGHT ABSORPTION, AND PIGMENT COMPOSITION OF MACROCYSTIS PYRIFERA (LAMINARIALES, PHAEOPHYCEAE) BLADES FROM DIFFERENT DEPTHS,JOURNAL OF PHYCOLOGY, Issue 6 2006María Florencia Colombo-Pallotta Macrocystis pyrifera (L.) C. Agardh is a canopy-forming species that occupies the entire water column. The photosynthetic tissue of this alga is exposed to a broad range of environmental factors, particularly related to light quantity and quality. In the present work, photosynthetic performance, light absorption, pigment composition, and thermal dissipation were measured in blades collected from different depths to characterize the photoacclimation and photoprotection responses of M. pyrifera according to the position of its photosynthetic tissue in the water column. The most important response of M. pyrifera was the enhancement of photoprotection in surface and near-surface blades. The size of the xanthophyll cycle pigment pool (XC) was correlated to the nonphotochemical quenching (NPQ) of chl a fluorescence capacity of the blades. In surface blades, we detected the highest accumulation of UV-absorbing compounds, photoprotective carotenoids, ,XC, and NPQ. These characteristics were important responses that allowed surface blades to present the highest maximum photosynthetic rate and the highest PSII electron transport rate. Therefore, surface blades made the highest contribution to algae production. In contrast, basal blades presented the opposite trend. These blades do not to contribute significantly to photosynthetate production of the whole organism, but they might be important for other functions, like nutrient uptake. [source] Comparison of two plant functional approaches to evaluate natural restoration along an old-field , deciduous forest chronosequenceJOURNAL OF VEGETATION SCIENCE, Issue 2 2009Isabelle Aubin Abstract Question: Are direct and indirect trait-based approaches similar in their usefulness to synthesize species responses to successional stages? Location: Northern hardwood forests, Québec, Canada (45°01,,45°08,N; 73°58,,74°21,W). Methods: Two different trait-based approaches were used to relate plant functional traits to succession on an old-field , deciduous forest chronosequence: (i) a frequently used approach based on co-occurrence of traits (emergent groups), and (ii) a new version of a direct functional approach at the trait level (the fourth-corner method). Additionally, we selected two different cut-off levels for the herb subset of the emergent group classification in order to test its robustness and ecological relevance. Results: Clear patterns of trait associations with stand developmental stages emerged from both the emergent group and the direct approach at the trait level. However, the emergent group classification was found to hide some trait-level differences such as a shift in seed size, light requirement and plant form along the chronosequence. Contrasting results were obtained for the seven or nine group classification of the herbaceous subset, illustrating how critical is the number of groups for emergent group classification. Conclusion: The simultaneous use of two different trait-based approaches provided a robust and comprehensive characterization of vegetation responses in the old-field , deciduous forest chronosequence. It also underlines the different goals as well as the limitations and benefits of these two approaches. Both approaches indicated that abandoned pastures of the northern hardwood biome have good potential for natural recovery. Conversion of these lands to other functions may lead to irremediable loss of biodiversity. [source] Use of the ,-function to estimate the skewness of species responsesJOURNAL OF VEGETATION SCIENCE, Issue 6 2003Branko Karad Abstract. Response of a species to an environmental variable may be modeled and predicted using a wide spectrum of different functions. Contrary to other functions (Gaussian, polynomial etc), all parameters of the ,-function are interpretable in ecological terms. However, computational difficulties in the determination of the ,-function parameters initiated controversial debates on the applicability and usefulness of this function in vegetation modelling and gradient analysis. We propose a simple algorithm for fitting the ,-function to observed data. Analytic properties of the algorithm (its ability to recover the known species responses along gradients) are tested using a series of simulated data. In most cases the algorithm correctly estimated parameters of the simulated responses. [source] A modified CRISIS-HSQC for band-selective IMPRESSMAGNETIC RESONANCE IN CHEMISTRY, Issue 2 2005Scott A. Bradley Abstract CRISIS (Compensation of Refocusing Inefficiency with Synchronized Inversion Sweep) is a powerful technique for obtaining multiplicity-edited HSQC spectra without compromising sensitivity. However, the stringent requirement for the duration of the CRISIS waveforms makes them unsuitable for other functions, such as band selection or IMPRESS (IMProved REsolution using Symmetrically Shifted pulses). We report here a modified CRISIS-gHSQC pulse sequence employing time-reversed 13C ,/2 EBURP-2 pulses. This IC-bs-gHSQC (IMPRESS-CRISIS-bs-gHSQC) sequence was found to be equally useful for acquiring multiplicity-edited, band-selective spectra individually or in tandem with IMPRESS. Remarkably, the latter provides multiple spectra in significantly less time and is the preferred approach when several crowded regions need to be assigned unambiguously. The use of adiabatic sweeps and the CRISIS pulses enable IC-bs-gHSQC to give better sensitivity than the original IMPRESS sequence for band-selective spectra. Copyright © 2005 John Wiley & Sons, Ltd. [source] In silico mining of EST databases for novel pre-implantation embryo-specific zinc finger protein genes,MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 3 2001Kong-Bung Choo Abstract Progress in the understanding of early mammalian embryo development has been severely hampered by scarcity of study materials. To circumvent such a constraint, we have developed a strategy that involves a combination of in silico mining of new genes from expressed sequence tags (EST) databases and rapid determination of expression profiles of the dbEST-derived genes using a PCR-based assay and a panel of cDNA libraries derived from different developmental stages and somatic tissues. We demonstrate that in a random sample of 49 independent dbEST-derived zinc finger protein genes mined from a mouse embryonic 2-cell cDNA library, more than three-quarters of these genes are novel. Examination of characteristics of the human orthologues derived from these mouse genes reveals that many of them are associated with human malignancies. Expression studies have further led to the identification of three novel genes that are exclusively expressed in mouse embryos before or up to the 8-cell stage. Two of the genes, designated 2czf45 and 2czf48 (2czf for 2 -cell zinc finger), are zinc finger protein genes coding for a RBCC protein with a RFP domain and a protein with three C2H2 fingers, respectively. The third gene, designated 2cpoz56, codes for a protein with a POZ domain that is often associated with zinc finger proteins. These three genes are candidate genes for regulatory or other functions in early embryogenesis. The strategy described in this report should generally be applicable to rapid and large-scale mining of other classes of rare genes involved in other biological and pathological processes. Mol. Reprod. Dev. 59:249,255, 2001. © 2001 Wiley-Liss, Inc. [source] Packaging reminiscences: some thoughts on controversial mattersPACKAGING TECHNOLOGY AND SCIENCE, Issue 4 2002Frank PaineArticle first published online: 24 JAN 200 Abstract Packaging is necessary in a world dominated by marketing-based economies, which rely on it for the safe delivery of profitable products. It is different from many other functions because packaging has two faces and each face demands a special marketing approach. Also necessary to appreciate is that in the very marketing of all products packaging plays a vital part, yet it is still often treated as a necessary evil and is often used on an expediency basis by product manufacturers. Even the packaging manufacturers themselves still fall into the trap of providing what is asked for instead of designing what is really needed. To describe packaging as in a world of its own is no pretension, for wherever natural or manufactured products are produced, packaging is needed to contain, preserve and protect them in the journey to the market place. Food, drink, clothing, light engineering goods, china and glass, medicines and household chemicals,in all those industries it is packaging made of paper and board, glass, plastic and metal that serves them. The importance of properly designed packaging lies in the fact that it must meet the need for protection of the product from the hazards of damage and deterioration. At the same time it must also provide identification and attractive presentation and meet the appropriate environmental criteria, Copyright © 2002 John Wiley & Sons, Ltd. [source] Emmprin (basigin/CD147): Matrix metalloproteinase modulator and multifunctional cell recognition molecule that plays a critical role in cancer progressionPATHOLOGY INTERNATIONAL, Issue 7 2006Kazuki Nabeshima Emmprin (basigin, CD147) is a cell surface glycoprotein that belongs to the immunoglobulin superfamily. It is highly expressed on the surface of tumor cells and stimulates adjacent fibroblasts or tumor cells to produce matrix metalloproteinases. Moreover, it has recently been shown that emmprin also stimulates expression of vascular endothelial growth factor and hyaluronan, which leads to angiogenesis and anchorage-independent growth/multidrug resistance, respectively. These findings have made emmprin an important molecule in tumor progression and, thus, more attractive as a target for antitumor treatment. However, other functions of emmprin, including as an activator of T cells, a chaperone for monocarboxylate transporters, a receptor for cyclophilin A and a neural recognition molecule, are also being identified in physiological and pathological conditions. Therefore, it is essential to develop specific means to control particular functions of emmprin, for which elucidation of each mechanism is crucial. This review will discuss the role of emmprin in tumor progression and recent advances in the molecular mechanisms of diverse phenomena regulated by emmprin. [source] | ||||||||||||||||||||||||||||||||||||||||