Two axes in platelet-derived growth factor signaling: tyrosine phosphorylation and reactive oxygen species

The tyrosine phosphorylation cascade is a hallmark of platelet-derived growth factor (PDGF)- induced signal transduction. The amplitude and propagation of the tyrosine phosphorylation signal relies on the balance between tyrosine kinase and tyrosine phosphatase. The tyrosine kinase is latent in the absence of stimulation, whereas the tyrosine phosphatase is highly and constitutively active. Therefore, the kinase activation should be accompanied by temporal and spatial inactivation of tyrosine phosphatase to achieve the robust amplification of tyrosine phosphorylation. For the past decade, reactive oxygen species have been receiving a great deal of attention with regard to their ability to shut down tyrosine phosphatase activities in a reversible manner. In this article, the crosstalk between tyrosine phosphorylation and reactive oxygen species in PDGF signaling is discussed. Received 2 October 2006; received after revision 13 November 2006; accepted 27 November 2006     

Structure and function of coagulogen,a clottable protein in horseshoe crabs

Mammalian blood coagulation is based on the proteolytically induced polymerization of fibrinogens. Initially, fibrin monomers noncovalently interact with each other. The resulting homopolymers are further stabilized when the plasma transglutaminase (TGase) intermolecularly cross-links -(-glutamyl)lysine bonds. In crustaceans, hemolymph coagulation depends on the TGase-mediated cross-linking of specific plasma-clotting proteins, but without the proteolytic cascade. In horseshoe crabs, the proteolytic coagulation cascade triggered by lipopolysaccharides and b-1,3-glucans leads to the conversion of coagulogen into coagulin, resulting in noncovalent coagulin homopolymers through head-to-tail interaction. Horseshoe crab TGase, however, does not cross-link coagulins intermolecularly. Recently, we found that coagulins are cross-linked on hemocyte cell surface proteins called proxins. This indicates that a cross-linking reaction at the final stage of hemolymph coagulation is an important innate immune system of horseshoe crabs.Received 27 October 2003; received after revision 25 November 2003; accepted 1 December 2003     

Decoding the Hedgehog signal in animal development

The Hedgehog (Hh) family of secreted proteins plays essential roles in a myriad of developmental processes via a complex signaling cascade conserved in species ranging from insects to mammals. In many developmental contexts, Hh acts as long-range morphogen to control distinct cellular outcomes as a function of its concentration. Here we review the current understanding of the Hh signaling mechanisms that govern the establishment of the Hh gradient and the transduction of the Hh signal with an emphasis on the intracellular signaling cascade from the receptor to the nuclear effector. We discuss how graded Hh signals are transduced to govern distinct developmental outcomes. Received 28 October 2005; received after revision 6 February 2006; accepted 15 February 2006     

Oxygen consumption of hibernating worker ants Dolichoderus quadripunctatus as a function of temperature and level of population]

The oxygen consumption of "hibernating" ants shows a grouping effect translated by a significant decrease in the level of oxygen consumption as a function of grouping effect. The general relation between oxygen consumption and the living body mass is the same, if the living material belongs to a single animal, or a social community of ants.     

β-phenylethyl isothiocyanate-mediated apoptosis in hepatoma HepG2 cells

-Phenylethyl isothiocyanate (PEITC) is a promising chemoprotective compound that is routinely consumed in the diet as its glucosinolate precursor. Previous studies have shown that PEITC can inhibit phase I enzymes and induce phase II detoxification enzymes along with apoptosis in vitro. The detailed mechanisms involved in the apoptotic cascade, however, have not been elucidated. In the present study, we demonstrate that PEITC can induce apoptosis in hepatoma HepG2 cells in a concentration- and time-dependant manner as determined by TUNEL positive and SubG1 population analysis. Caspase-3-like activity and poly(ADP-ribosyl)polymerase cleavage increased during treatment with 20 µM PEITC; high concentrations, however, induced necrosis. Pre-treatment with Z-VAD-FMK and the caspase-3-specific inhibitor Ac-DEVD-CHO prevented PEITC-induced apoptosis, as determined by caspase-3-like activity and DNA fragmentation. Additional investigations also showed that at concentrations of 5-C10 µM PEITC, DNA synthesis was inhibited and G2/M phase cell cycle arrest occurred, correlating with an alteration in cyclin B1 and p34^cdc2 protein levels. Furthermore, we also demonstrate a concentration- and time-dependant burst of superoxide (O₂^-) in PEITC-treated cells. However, pre- and co-treatment with the free radical scavengers Trolox, ascorbate, mannitol, uric acid and the superoxide mimetic manganese (III) tetrakis (N-methyl-2-pyridyl) porphyrin failed to prevent PEITC-mediated apoptosis. Taken together, these results suggest that PEITC potently induces apoptosis and cell cycle arrest in HepG2 cells and that the generation of reactive oxygen species appears to be a secondary effect.Received 23 December 2002; accepted 22 April 2003     

MAP kinase signalling: interplays between plant PAMP- and effector-triggered immunity

In plants, mitogen-activated protein kinase (MAPK) cascades are involved in regulating many biological processes including immunity. They relay signals from membrane-residing immune receptors to downstream components for defense activation. Arabidopsis MPK3/6 and MPK4 are activated in two parallel MAPK cascades during PAMP-triggered immunity. MPK3/6 have been implicated in the activation of various immune responses and their inactivation leads to compromised defense against pathogens. On the other hand, the MEKK1-MKK1/2-MPK4 cascade plays critical roles in basal resistance. Disruption of this MAPK cascade results in constitutive defense responses mediated by the NB-LRR protein SUMM2. Interestingly, SUMM2 guards the MEKK1-MKK1/2-MPK4 cascade activity indirectly through monitoring the phosphorylation status of CRCK3, which is a substrate of MPK4. From the pathogens’ side, a number of effectors are shown to target various components of MAPK cascades in plants. Inactivation of MPK4 by the Pseudomonas effector HopAI1 triggers SUMM2-mediated immunity. Together, these findings suggest intricate interplays between PAMP-triggered immunity and effector-triggered immunity via MAPK signaling.     

A nonenzymic oxygen uptake and its implications in the assay of 4-hydroxyphenylpyruvate dioxygenase by an oxygen electrode

Summary The oxygen electrode provides a rapid, convenient assay for 4-hydroxyphenylpyruvate dioxygenase. However, due to the occurrence of a nonenzymic oxygen consumption when reducing agents were mixed witha,a-dipyridyl, its use is restricted to species which do not require reducing agents or necessitates the addition of catalase to the reaction mixture.     

Neuroglobin,seven years after

Neuroglobin is expressed in vertebrates brain and belongs to a branch of the globin family that diverged early in evolution. Sequence conservation suggests a relevant role in the nervous system, with tight structural restraints. Experiments in vivo and in vitro showed increased hypoxic stress damage upon repressing neuroglobin biosynthesis and improved recovery following overexpression. Neuroglobin shows internal heme hexacoordination, which controls oxygen affinity and kinetics. Neuroglobin concentration, oxygen affinity and enhanced autooxidation question a role in oxygen delivery; thus it was proposed that the neuroprotective effect might be due to radical scavenging or activation of protection mechanisms. Neuroglobin's structure shows a peculiar internal cavity of very large size. Binding of heme ligands is associated to a conformational change involving the heme that "slides" into the pre-existing cavity and makes the sixth coordination position available. These features may pave the way to an understanding of neuroprotection by neuroglobin.     

Non-canonical activation of Notch signaling/target genes in vertebrates

Evolutionarily conserved Notch signaling orchestrates diverse physiological mechanisms during metazoan development and homeostasis. Classically, ligand-activated Notch receptors transduce the signaling cascade through the interaction of DNA-bound CBF1-co-repressor complex. However, recent reports have demonstrated execution of a CBF1-independent Notch pathway through signaling cross-talks in various cells/tissues. Here, we have tried to congregate the reports that describe the non-canonical/CBF1-independent Notch signaling and target gene activation in vertebrates with specific emphasis on their functional relevance.     

Die physikalische Kieme der Wasserinsekten

Summary The air stores carried by a number of aquatic insects have: a) a hydrostatic function (Brocher, Oortwijn-Botjes, Thorpe, andCrisp); b) the function of an oxygen store (Ege, de Ruiter et al.) and c) the function of a physical gill (Strauss-Durckheim, Ege, etc.). The fact that oxygen is taken up from water with the aid of an air bubble was demonstrated forNotonecta by comparing the life time of insects with and without physical gill (while replenishing the oxygen store from the air was prevented) byEge, and forCorixa byPopham, whileVlasblom determined the oxygen uptake from water with and without air bubble forNotonecta, Naucoris, Corixa, Sigara andNepa. Nepa andSigara can take up considerable quantities of oxygen by cutaneous respiration.During the summer, the gill function of the air store ofNotonecta andNaucoris is of importance only when a water current passes along the animal, caused by ventilation movements of the legs (de Ruiter et al.). At low temperatures, however, the metabolic rate is so low that in many instances the physical gill provides the oxygen required without ventilation movements.An apparatus for the simultaneous determination of oxygen uptake from air and water (Wolvekamp andVlasblom) gave results that provided a means of evaluating the importance of the physical gill function.In some cases, the air store, although in direct contact with the water, does not need to be replenished. InAphelocheirus andElmis, the negative pressure in the bubble, caused by oxygen consumption and the diffusing out of part of the nitrogen, is compensated for by the mechanical resistance of a feltwork of thin hairs and the surface tension of the boundary layer of the water (Thorpe andCrisp). In the African beetle,Potamodytes, the unprotected air bubble is permanent because the strong river currents produce a lowered pressure around the animal according toBernoulli's principle (Stride).     

Melatonin biosynthesis in the mammalian pineal gland

Summary Rhythmic production of melatonin by the mammalian pineal occurs in response to noradrenergic stimulation which produces a cascade of biochemical events within the pinealocyte. In the rat, massive changes in NAT activity result from an increase in intracellular c-AMP levels produced by a synergistic interaction whereby an ₁ activation amplifies -adrenergic stimulation. The intracellular events mediating this effect are described. A major aspect of the temporal control of melatonin production is the programmed down-regulation of responses to noradrenergic stimulation once the initial surge of c-AMP is produced. Noradrenergic activation of the gland also influences other enzymic functions, including tryptophan hydroxylase and HIOMT activities, and produces a dramatic increase in intracellular c-GMP levels. Other neurotransmitters and neuropeptides, e.g. VIP, may also influence pineal function and comparisons are, made between the rat, the subject of the bulk of experimental studies, and other species.     

Skeletal muscle secretome in Duchenne muscular dystrophy: a pivotal anti-inflammatory role of adiponectin

Background

Persistent inflammation exacerbates the progression of Duchenne muscular dystrophy (DMD). The hormone, adiponectin (ApN), which is decreased in the metabolic syndrome, exhibits anti-inflammatory properties on skeletal muscle and alleviates the dystrophic phenotype of mdx mice. Here, we investigate whether ApN retains its anti-inflammatory action in myotubes obtained from DMD patients. We unravel the underlying mechanisms by studying the secretome and the early events of ApN.

Methods

Primary cultures of myotubes from DMD and control patients were treated or not by ApN after an inflammatory challenge. Myokines secreted in medium were identified by cytokine antibody-arrays and ELISAs. The early events of ApN signaling were assessed by abrogating selected genes.

Results

ApN retained its anti-inflammatory properties in both dystrophic and control myotubes. Profiling of secretory products revealed that ApN downregulated the secretion of two pro-inflammatory factors (TNFα and IL-17A), one soluble receptor (sTNFRII), and one chemokine (CCL28) in DMD myotubes, while upregulating IL-6 that exerts some anti-inflammatory effects. These changes were explained by pretranslational mechanisms. Earlier events of the ApN cascade involved AdipoR1, the main receptor for muscle, and the AMPK-SIRT1-PGC-1α axis leading, besides alteration of the myokine profile, to the upregulation of utrophin A (a dystrophin analog).

Conclusion

ApN retains its beneficial properties in dystrophic muscles by activating the AdipoR1-AMPK-SIRT1-PGC-1α pathway, thereby inducing a shift in the secretion of downstream myokines toward a less inflammatory profile while upregulating utrophin. ApN, the early events of the cascade and downstream myokines may be therapeutic targets for the management of DMD.

     

Scaling of respiratory areas in relation to oxygen consumption of vertebrates

Summary Resting oxygen consumption is generally related to the 0.75 power of the body mass. More recent studies on the morphometry of the lungs and gills of vertebrates indicate that the respiratory surfaces increase in area more closely to M^1.0. A possible explanation is that the gas exchange surface is more to be related to themaximum oxygen uptake capacity, rather than to the resting condition. However, recent studies, especially in African mammals, suggest that scales to a power similar to that for resting oxygen consumption and hence cannot be directly related to the respiratory surfaces.In fishes the situation seems to be different as appears to increase more rapidly than standard . Consequently, the ratio between these two is greater in larger specimens and hence they have a greater scope for activity. A possible interpretation of this apparent difference may be related to differences in ventilation, as the tidal ventilation of lungs can lead to a reduction in the difference in oxygen pressure across the exchange surface, whereas in fish gills with increasing size and increasing water velocity there would be a reduction in the resistance to gas transfer. However, it is important to appreciate that such generalizations probably do not hold in all cases and the position of intermediate groups of vertebrates needs to be more closely assessed.This paper is based upon part of a lecture given at the Conference of Comparative Respiratory Physiology, Göttingen, BRD, August 1982.     

Oxygen consumption in rat skeletal muscle at various rates of oxygen delivery

Summary Resting oxygen consumption of the vascularly isolated gracilis anticus muscle of the rat perfused by natural circulation via the femoral artery with diluted or undiluted blood depends on oxygen delivery (the product of flow rate and arterial oxygen concentration-DO₂) only when DO₂ falls below 16 l/min/g.     

The effects of physostigmine on the oxygen uptake in rat brain tissue

Summary Physostigmine in a dose of 0.1 mg/kg i.v. expressly stimulated the oxygen uptake in the rat cerebral cortex. This effect was blocked by propranolol and seems be mediated by catecholamines. Since atropine also antagonized the stimulant effect of physostigmine, it appears that the action of physostigmine is primarily cholinergic and that the adrenergic effect is a secondary phenomenon. The higher dose of physostigmine (0.4 mg/kg i.v.) caused a depression of rat brain oxygen uptake.This work was supported by a grant from Serbian Republic Scientific Fund (ZMNU SR Srbija), Belgrade (Yugoslavia).The authors wish to acknowledge the skilful technical assistance of Lj. Krsti.     

Restoration of the poor oxygen transport function of ACD-stored blood by pyridoxal 5′-phosphate

Summary Pyridoxal 5-phosphate is easily incorporated into ACD-stored erythrocytes without decrease of ATP, and restores the poor oxygen transport function with a similar effect to 2,3-diphosphoglycerate.     

The effect of camphor on mitochondrial respiration

Summary Camphor at <8 moles/mg protein reduced the rate of oxygen consumption by rat liver mitochondria. The effect occurs only with NAD⁺-linked substrates. Succinate linked respiration was inhibited but this appears to be caused by some conversion of succinate to malate. At higher levels, camphor increases oxygen consumption with succinate substrate, by uncoupling at site II.Acknowledgment. D.F.G.-C. is grateful to the New Zealand Cancer Society (Wellington Branch), for financial support. We thank Mrs E. Dye for technical assistance and Dr W. Jordan for valuable discussions.     

The family of iron responsive RNA structures regulated by changes in cellular iron and oxygen

The life of aerobes is dependent on iron and oxygen for efficient bioenergetics. Due to potential risks associated with iron/oxygen chemistry, iron acquisition, concentration, storage, utilization, and efflux are tightly regulated in the cell. A central role in regulating iron/oxygen chemistry in animals is played by mRNA translation or turnover via the iron responsive element (IRE)/iron regulatory protein (IRP) system. The IRE family is composed of three-dimensional RNA structures located in 3′ or 5′ untranslated regions of mRNA. To date, there are 11 different IRE mRNAs in the family, regulated through translation initiation or mRNA stability. Iron or oxidant stimuli induce a set of graded responses related to mRNA-specific IRE substructures, indicated by differential responses to iron in vivo and binding IRPs in vitro. Molecular effects of phosphorylation, iron and oxygen remain to be added to the structural information of the IRE-RNA and IRP repressor in the regulatory complex. Received 21 April 2007; received after revision 13 July 2007; accepted 2 August 2007     

Biochemistry,evolution and physiological function of the Rnf complex,a novel ion-motive electron transport complex in prokaryotes

Microbes have a fascinating repertoire of bioenergetic enzymes and a huge variety of electron transport chains to cope with very different environmental conditions, such as different oxygen concentrations, different electron acceptors, pH and salinity. However, all these electron transport chains cover the redox span from NADH + H⁺ as the most negative donor to oxygen/H₂O as the most positive acceptor or increments thereof. The redox range more negative than −320 mV has been largely ignored. Here, we have summarized the recent data that unraveled a novel ion-motive electron transport chain, the Rnf complex, that energetically couples the cellular ferredoxin to the pyridine nucleotide pool. The energetics of the complex and its biochemistry, as well as its evolution and cellular function in different microbes, is discussed.