The superoxide-generating NADPH oxidase: structural aspects and activation mechanism

Flavocytochrome b ₅₅₈ is the catalytic core of the respiratory-burst oxidase, an enzyme complex that catalyzes the NADPH-dependent reduction of O₂ into the superoxide anion O₂ ^- in phagocytic cells. Flavocytochrome b ₅₅₈ is anchored in the plasma membrane. It is a heterodimer that consists of a large glycoprotein gp91phox (phox for phagocyte oxidase) (β subunit) and a small protein p22phox (α subunit). The other components of the respiratory-burst oxidase are water-soluble proteins of cytosolic origin, namely p67phox, p47phox, p40phox and Rac. Upon cell stimulation, they assemble with the membrane-bound flavocytochrome b ₅₅₈ which becomes activated and generates O₂ ^-. A defect in any of the genes encoding gp91phox, p22phox, p67phox or p47phox results in chronic granulomatous disease, a genetic disorder characterized by severe and recurrent infections, illustrating the role of O₂ ^- and the derived metabolites H₂O₂ and HOCl in host defense against invading microorganisms. The electron carriers, FAD and hemes b, and the binding site for NADPH are confined to the gp91phox subunit of flavocytochrome b ₅₅₈ . The p22phox subunit serves as a docking site for the cytosolic phox proteins. This review provides an overview of current knowledge on the structural organization of the O₂ ^--generating flavocytochrome b ₅₅₈ , its kinetics, its mechanism of activation and the regulation of its biosynthesis. Homologues of gp91phox, called Nox and Duox, are present in a large variety of non-phagocytic cells. They exhibit modest O₂ ^--generating oxidase activity, and some act as proton channels. Their role in various aspects of signal transduction is currently under investigation and is briefly discussed. Received 28 May 2002; received after revision 20 June 2002; accepted 24 June 2002     

4-Hydroxynonenal-modified amyloid-beta peptide inhibits the proteasome: possible importance in Alzheimer's disease

The amyloid β-peptide (Aβ) is a 4-kDa species derived from the amyloid precursor protein, which accumulates in the brains of patients with Alzheimer’s disease. Although we lack full understanding of the etiology and pathogenesis of selective neuron death, considerable data do imply roles for both the toxic Aβ and increased oxidative stress. Another significant observation is the accumulation of abnormal, ubiquitin-conjugated proteins in affected neurons, suggesting dysfunction of the proteasome proteolytic system in these cells. Recent reports have indicated that Aβ can bind and inhibit the proteasome, the major cytoslic protease for degrading damaged and ubiquitin-conjugated proteins. Earlier results from our laboratory showed that moderately oxidized proteins are preferentially recognized and degraded by the proteasome; however, severely oxidized proteins cannot be easily degraded and, instead, inhibit the proteasome. We hypothesized that oxidatively modified Aβ might have a stronger (or weaker) inhibitory effect on the proteasome than does native Aβ. We therefore also investigated the proteasome inhibitory action of Aβ _1–40 (a peptide comprising the first 40 residues of Aβ) modified by the intracellular oxidant hydrogen peroxide, and by the lipid peroxidation product 4-hydroxynonenal (HNE). H₂O₂ modification of Aβ _1–40 generates a progressively poorer inhibitor of the purified human 20S proteasome. In contrast, HNE modification of Aβ _1–40 generates a progressively more selective and efficient inhibitor of the degradation of fluorogenic peptides and oxidized protein substrates by human 20S proteasome. This interaction may contribute to certain pathological manifestations of Alzheimer’s disease Received 26 September 2000; accepted 26 September 2000     

Protease-activated-receptor-2 affects protease-activated-receptor-1-driven breast cancer

Mammalian protease-activated-receptor-1 and -2 (PAR₁ and PAR₂) are activated by proteases found in the flexible microenvironment of a tumor and play a central role in breast cancer. We propose in the present study that PAR₁ and PAR₂ act together as a functional unit during malignant and physiological invasion processes. This notion is supported by assessing pro-tumor functions in the presence of short hairpin; shRNA knocked-down hPar2 or by the use of a truncated PAR₂ devoid of the entire cytoplasmic tail. Silencing of hPar2 by shRNA-attenuated thrombin induced PAR₁ signaling as recapitulated by inhibiting the assembly of Etk/Bmx or Akt onto PAR₁-C-tail, by thrombin-instigated colony formation and invasion. Strikingly, shRNA-hPar2 also inhibited the TFLLRN selective PAR₁ pro-tumor functions. In addition, while evaluating the physiological invasion process of placenta extravillous trophoblast (EVT) organ culture, we observed inhibition of both thrombin or the selective PAR₁ ligand; TFLLRNPNDK induced EVT invasion by shRNA-hPar2 but not by scrambled shRNA-hPar2. In parallel, when a truncated PAR₂ was utilized in a xenograft mouse model, it inhibited PAR₁–PAR₂-driven tumor growth in vivo. Similarly, it also attenuated the interaction of Etk/Bmx with the PAR₁-C-tail in vitro and decreased markedly selective PAR₁-induced Matrigel invasion. Confocal images demonstrated co-localization of PAR₁ and PAR₂ in HEK293T cells over-expressing YFP-hPar2 and HA-hPar1. Co-immuno-precipitation analyses revealed PAR₁-PAR₂ complex formation but no PAR₁-CXCR4 complex was formed. Taken together, our observations show that PAR₁ and PAR₂ act as a functional unit in tumor development and placenta-uterus interactions. This conclusion may have significant consequences on future breast cancer therapeutic modalities and improved late pregnancy outcome.     

The birth of modern science: culture, mentalities and scientific innovation

In a recent paper, Luc Faucher and others have argued for the existence of deep cultural differences between ‘Chinese’ and ‘East Asian’ ways of understanding the world and those of ‘ancient Greeks’ and ‘Americans’. Rejecting Alison Gopnik’s speculation that the development of modern science was driven by the increasing availability of leisure and information in the late Renaissance, they claim instead—following Richard Nisbett—that the birth of mathematical science was aided by ‘Greek’, or ‘Western’, cultural norms that encouraged analytic, abstract and rational theorizing. They argue that ‘Chinese’ and ‘East Asian’ cultural norms favoured, by contrast, holistic, concrete and dialectical modes of thinking. After clarifying some of the things that can be meant by ‘culture’ and ‘mentality’, the present paper shows that Faucher and his colleagues make a number of appeals—to the authority of comparative studies and history of science, to the psychological studies of Nisbett and his colleagues, and to a hidden assumption of strong cultural continuity in the West. It is argued that every one of these appeals is misguided, and, further, that the psychological findings of Nisbett and others have little bearing on questions concerning the origins of modern science. Finally, it is suggested that the ‘Needham question’ about why the birth of modern science occurred in Europe rather than anywhere else is itself multiply confused to the extent that it may express no significant query.     

Zur Stratigraphie der säugetierführenden Schichten von Java

Summary According to recent geological and paleontological investigations thePithecanthropus-beds of Java are pleistocene in age. The examinations of the fossil mammalian faunæ of the island are of utmost importance for this age-determination and the following faunæ can be recognized:Tji Djolang-fauna: Middle Pliocene.Kali Glagah-fauna: Upper Pliocene.Djetis-fauna withPithecanthropus: Lower Pleistocene.Trinil-fauna withPithecanthropus: Middle Pleistocene.Ngandong-fauna withHomo neanderthalensis soloensis: Upper Pleistocene.Sampoeng-fauna with a primitive type of the Wedda-Dravida-Australoid group: Subrecent. — Micropaleontological examinations of smallerForaminifera from marine equivalents of thePithecanthropus-beds confirm the age-determinations based on mammalia. — Recent field investigations prove that the lower and middle pleistocenePithecanthropus-beds are distinctlyfolded by a post-middle-pleistocene movement; the upper pleistocene Ngandong-terraces are not folded at all.     

The effect of a single dose of N-ethyl-N-nitrosourea on the fine structure of the brain of the rat

Résumé Une forte dose de composé carcinogène N-éthyle-N-nitrosourée (ENU) a été injectée à des rats Wistar mâles adultes: des changements ultrastructuraux des cerveaux furent observés. La dégénérescence et la nécrose des oligodendrocytes, l'oedème des astrocytes et l'accroissement du nombre des corps denses dans les cellules gliales ont été les réactions du cerveau à ce stimulus chimique.

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I wish to thank Dr.M. Kremer, Director of the Department of Neurological Studies; and Drs.A. R. Lieberman andP. Kleihues and also Mr.A. L. E. Barron for the photographic work. I am particularly indebted to Dr.Helen Grant and the staff of the Department of Neuropathology, Bland-Sutton Institute, for their continual help and advice.     

Distribution du système glio-interstitiel chez les Mollusques: Les parties tonique et phasique du muscle adducteur postérieur chezAnodonta

Summary The structure and nervous supply of the tonic and the phasic parts of the posterior adductor muscle ofAnodonta were studied by electron microscopy. A teloglia accompanies the nerves of the tonic part, while this tissue is absent from the phasic part of the muscle. This distribution is compared to the parallel situation in marine gastropods; and it is tentatively proposed that the development of the gliointerstitial tissue in the muscle of a mollusc depends on the type of contraction of this muscles and/or the development of the extracellular spaces.

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Ce travail a été réalisé au sein de l'Equipe de Recherche associée au C.N.R.S. No. 183 et au Centre de Microscopie Electronique Appliquée à la Biologie de l'Université Claude Bernard.     

Inhibition sélective des «vraies» et «pseudo»-cholinestérases du rectus de grenouille

Summary The hydrolysis of acetylcholine chloride (0.01M) by frog's rectus extracts, is inhibited by low concentrations of 3318 CT (CI-50 3.2×10^–7) and high concentrations of D.F.P. (CI-50 1.3×10^–5). Inversely, the hydrolysis of butyrylcholine perchlorate is inhibited by low concentrations of D.F.P. (CI-50 3×10^–9) and high concentrations of 3318 CT (CI-50 3×10^–4). Both are inhibited by similar concentrations of neostigmine (CI-50 1.1×10^–7 and 1.5×10^–7). Frog's rectus thus contains true and pseudo-cholinesterases. The inhibitions produced by D.F.P. added to the muscle itself (and not the extract) correlates well with the potentiation of the corresponding ester. Sensitization to AcCh and to BuCh appears to be specifically related to the inhibition of Ac ChE for the former ester, of XChE for the second one.     

Announcement of the Editor-in-Chief

Announcement.  As decided in July 2005 we continue to publish once a year (in July) the names of the authors and the titles of the two most read (by Internet) Research Papers and Reviews published in Cell. Mol. Life Sci. the previous year. Thus we have the pleasure to provide you with the results of 2005. Research Articles  

Einstein,Nordström and the early demise of scalar,Lorentz-covariant theories of gravitation

Conclusion The advent of the general theory of relativity was so entirely the work of just one person — Albert Einstein — that we cannot but wonder how long it would have taken without him for the connection between gravitation and spacetime curvature to be discovered. What would have happened if there were no Einstein? Few doubt that a theory much like special relativity would have emerged one way or another from the researchers of Lorentz, Poincaré and others. But where would the problem of relativizing gravitation have led? The saga told here shows how even the most conservative approach to relativizing gravitation theory still did lead out of Minkowski spacetime to connect gravitation to a curved spacetime. Unfortunately we still cannot know if this conclusion would have been drawn rapidly without Einstein's contribution. For what led Nordström to the gravitational field dependence of lengths and times was a very Einsteinian insistence on just the right version of the equality of inertial and gravitational mass. Unceasingly in Nordström's ear was the persistent and uncompromising voice of Einstein himself demanding that Nordström see the most distant consequences of his own theory.     

Explaining Leibniz equivalence as difference of non-inertial appearances: Dis-solution of the Hole Argument and physical individuation of point-events

“The last remnant of physical objectivity of space–time” is disclosed in the case of a continuous family of spatially non-compact models of general relativity (GR). The physical individuation of point-events is furnished by the autonomous degrees of freedom of the gravitational field (viz., the Dirac observables) which represent—as it were—the ontic part of the metric field. The physical role of the epistemic part (viz. the gauge variables) is likewise clarified as embodying the unavoidable non-inertial aspects of GR. At the end the philosophical import of the Hole Argument is substantially weakened and in fact the Argument itself dissolved, while a specific four-dimensional holistic and structuralist view of space–time (called point-structuralism) emerges, including elements common to the tradition of both substantivalism and relationism. The observables of our models undergo real temporal change: this gives new evidence to the fact that statements like the frozen-time character of evolution, as other ontological claims about GR, are model dependent.     

Sonnenflecken und ihre terrestrischen Wirkungen

Summary After a discussion of the 11-year solar-cycle as regarded from the standpoint of the eruption-hypothesis, which offers a possibility to predict the solar activity for several years and after a review of the magnetic properties of the sun and the sunspots, the paper deals with the new theories of the spots and the solar-cycle as suggested byAlfvén andWalén.The terrestrial effects of the phenomena associated with the solar cycle are classified into 4 groups: effects produced by a) a wave radiationW _k emitted continuously by the sun, b) a wave radiationW _e emitted from the chromospheric eruptions, c) a particle radiationP _k emitted by the so-called M-regions, and d) a particle radiationP _e ejected from the eruptions. The connection between the solar eruptions and the radiationsP _e ,W _e is a well established fact; on the other hand the radiationsW _k ,P _k could be connected by the author with the solar corona and the stationary solar prominences respectively.To account for the intensity of theW _k -radiation a temperature of the solar corona of one million degree is required in agreement with the observed temperature. The heating of the corona occurs in the electric field around an increasing sunspot. As in the corona the mean free path amounts to several kilometers, particles may be accelerated up to 1000 eV, so far the conditionh=0 is fulfilled. Generally speaking acceleration is possible only in such regions whereh andh are not perpendicular to each other.     

Obtention d'un dérivé du sclaréol à partir de l'ambréine

Summary One of the products of the chromic acid oxidation of the animal triterpene ambreine (V) has been identified with the lactone C₁₆H₂₆O₂ (III) whichRuzicka and coll. had isolated by oxidation of the plant diterpene sclareol (II). This identification confirms recent findings of the authors and ofRuzicka, Dürst andJeger and shows that the hydroxyle of ambreine (V) is at the same place as the hydroxyle of sclareol which is attached to the perhydronaphtalene ring.

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4^me communication sur les constituants de l'ambre gris     

Variance to mean ratio and the spatial distribution of animals

Summary Biological and statistical aspects of the application of variance to mean ratio to describe spatial distribution of animals are discussed. It is shown that the parameterb inTaylor's power laws ²=a m ^b shows intra-specific variation depending on the distribution of the constituent units of the population.a andb are only parameters of a very empirical way of describing the relation between variance and mean, which itself is an indicator for spatial distribution. Hence,a andb depend on the distribution behaviour of the animals, and not vice versa.Acknowledgments: The work was developed whilst I was at the Department of Entomology, University of Nairobi. I am grateful to ProfessorT. R. Odhiambo for his interest. I thank the referee for comments.     

Effect of various solubilizers on angiotensinII receptors in bovine adrenocortical plasma membranes

Summary 3 monionic detergents, Triton X-100, Lubrol WX and NP-40, inhibited binding of [³H]-AT_II to bovine adrenocortical plasma membranes. This effect appeared to be direct and not due to solubilization of the AT_II receptor by these agents. Sodium deoxycholate and the chaotropic ions, ClO ₄ ^– and Br^–, produced effects similar to the nonionic detergents.Supported by grants from the Quebec Heart Foundation and the Medical Research Council of Canada. The authors acknowledge the technical assistance of Mrs D. Michaud and the secretarial aide of Mrs D. Huot-Blais, and Miss L. Leblanc.     

Characterization of the estrogen receptors in the uterine and blood eosinophil leukocytes

Summary Estrogen receptors are found in the rat uterine and in the eosinophil-rich human blood leukocyte 24,000g fractions, but not in the low-eosinophil count human blood leukocyte 24,000g fraction. The total number of binding sites per blood eosinophil leukocyte is 7,400 sites per cell, and theK _d =5.6×10^–10 M.Acknowledgments. This work was supported in part by funds from the Unité de Recherches sur le Métabolisme Moléculaire et la Physio-Pathologie des Stéroïdes de l'INSERM, Hôpital de Bicetre, France, and by grant No. 2015 from the Oficina Técnica de Desarrollo Científico y Creación Artística of the University of Chile.     

Redundant and unique roles of coronin proteins in <Emphasis Type="Italic">Dictyostelium</Emphasis>

Dictyostelium discoideum harbors a short (CRN12) and a long coronin (CRN7) composed of one and two beta-propellers, respectively. They are primarily present in the cell cortex and cells lacking CRN12 (corA ⁻) or CRN7 (corB ⁻) have defects in actin driven processes. We compared the characteristics of a mutant cell line (corA ⁻ /corB ⁻) lacking CRN12 and CRN7 with the single mutants focusing on cytokinesis, phagocytosis, chemotaxis and development. Cytokinesis, uptake of small particles, and developmental defects were not enhanced in the corA ⁻ /corB ⁻ strain as compared to the single mutants, whereas motility and phagocytosis of yeast particles were more severely impaired. It appears that although both proteins affect the same processes they do not act in a redundant manner. Rather, they often act antagonistically, which is in accordance with their proposed roles in the actin cytoskeleton where CRN12 acts in actin disassembly whereas CRN7 stabilizes actin filaments and protects them from disassembly.     

Poincaréan intuition revisited: what can we learn from Kant and Parsons?

This paper provides a comprehensive critique of Poincaré’s usage of the term intuition in his defence of the foundations of pure mathematics and science. Kant’s notions of sensibility and a priori form and Parsons’s theory of quasi-concrete objects are used to impute rigour into Poincaré’s interpretation of intuition. In turn, Poincaré’s portrayal of sensible intuition as a special kind of intuition that tolerates the senses and imagination is rejected. In its place, a more harmonized account of how we perceive concrete objects is offered whereby intuitive knowledge is consistently a priori whatever the domain of application.     

Altered expression of securin (Pttg1) and serpina3n in the auditory system of hearing-impaired Tff3-deficient mice

Introduction

Tff3 peptide exerts important functions in cytoprotection and restitution of the gastrointestinal (GI) tract epithelia. Moreover, its presence in the rodent inner ear and involvement in the hearing process was demonstrated recently. However, its role in the auditory system still remains elusive. Our previous results showed a deterioration of hearing with age in Tff3-deficient animals.

Results

Present detailed analysis of auditory brain stem response (ABR) measurements and immunohistochemical study of selected functional proteins indicated a normal function and phenotype of the cochlea in Tff3 mutants. However, a microarray-based screening of tissue derived from the auditory central nervous system revealed an alteration of securin (Pttg1) and serpina3n expression between wild-type and Tff3 knock-out animals. This was confirmed by qRT-PCR, immunostaining and western blots.

Conclusions

We found highly down-regulated Pttg1 and up-regulated serpina3n expression as a consequence of genetically deleting Tff3 in mice, indicating a potential role of these factors during the development of presbyacusis.