Different carbon sources affect lifespan and protein redox state during Saccharomyces cerevisiae chronological ageing

In this study, a proteomic approach that combines selective labelling of proteins containing reduced cysteine residues with two-dimensional electrophoresis/mass spectrometry was used to evaluate the redox state of protein cysteines during chronological ageing in Saccharomyces cerevisiae. The procedure was developed on the grounds that biotinconjugated iodoacetamide (BIAM) specifically reacts with reduced cysteine residues. BIAM-labelled proteins can then be selectively isolated by streptavidin affinity capture. We compared cells grown on 2% glucose in the exponential phase and during chronological ageing and we found that many proteins undergo cysteine oxidation. The target proteins include enzymes involved in glucose metabolism. Both caloric restriction and growth on glycerol resulted in a decrease in the oxidative modification. Furthermore, in these conditions a reduced production of ROS and a more negative glutathione half cell redox potential were observed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Received 15 September 2008; received after revision 17 December 2008; accepted 06 January 2009     

Enhanced susceptibility of T lymphocytes to oxidative stress in the absence of the cellular prion protein

The cellular prion glycoprotein (PrP^C) is ubiquitously expressed but its physiologic functions remain enigmatic, particularly in the immune system. Here, we demonstrate in vitro and in vivo that PrP^C is involved in T lymphocytes response to oxidative stress. By monitoring the intracellular level of reduced glutathione, we show that PrP^−/− thymocytes display a higher susceptibility to H₂O₂ exposure than PrP^+/+ cells. Furthermore, we find that in mice fed with a restricted diet, a regimen known to increase the intracellular level of ROS, PrP^−/− thymocytes are more sensitive to oxidative stress. PrP^C function appears to be specific for oxidative stress, since no significant differences are observed between PrP^−/− and PrP^+/+ mice exposed to other kinds of stress. We also show a marked evolution of the redox status of T cells throughout differentiation in the thymus. Taken together, our results clearly ascribe to PrP^C a protective function in thymocytes against oxidative stress.     

Über ein Chromoplastin aus reifenden Tomaten

Summary (1) By precipitation with ammonium sulfate, streptomycin, or calcium salts, we obtained from the pulp of tomatoes a substance containing carotinoids. The behaviour of this substance was analogous to that of chloroplst-substance and to that of animal cytoplasmatic nucleoproteins. Like these it contains proteins, lipids, and very probably nucleic acids. We regard this substance as achromoplastine.(2) Experiments with slices of green tomatoes show that the changing over of the chlorophyll content into the carotinoid content is inhibited by the presence of streptomycin.(3) Streptomycin inhibits the formation of chlorophyll in etiolated separated cabbage leafs, just as this drug inhibits the formation of chlorophyll in growing seeds.(4) The development of anthocyanides is not influenced by streptomycin.     

Peroxiredoxin 2 (<Emphasis Type="Italic">PRDX2</Emphasis>), an antioxidant enzyme,is underexpressed in Down syndrome fetal brains

Suppression subtractive hybridization performed on Down syndrome (DS) versus control fetal brains revealed differential expression of peroxiredoxin 2 (PRDX2), mapped at 13q12. Peroxiredoxins are antioxidant enzymes involved in protein and lipid protection against oxidative injury and in cellular signalling pathways regulating apoptosis. The under-expression of PRDX2 observed in DS samples was confirmed by realtime PCR (0.73-fold). To test whether decreased expression is associated with enhanced sensitivity of DS neurons to reactive oxygen species, we down-regulated PRDX2 through stable transfections of SH-SY5Y neuroblastoma cells with antisense contructs of the complete PRDX2 coding sequence. In addition, we over-expressed SOD1 and compared the effects of the two genes on cell viability. Cells transfected with either construct showed similar sensitivity to oxidative stress in addition to increased apoptosis under basal conditions and after treatment with oxidative cytotoxic agents. This suggests that the decreased expression of PRDX2 may contribute to the altered redox state in DS at levels comparable to that of the increased expression of SOD1.Received 4 February 2003; received after revision 31 March 2003; accepted 25 April 2003     

Neuroprotective effects of <Emphasis Type="Italic">Ginkgo biloba</Emphasis> extract

Ginkgo biloba extract has been therapeutically used for several decades to increase peripheral and cerebral blood flow as well as for the treatment of dementia. The extract contains multiple compounds such as flavonoids and terpenoids that are thought to contribute to its neuroprotective and vasotropic effects. In this review, we summarize the experimental results on the mechanism of neuroprotection induced by standardized extract of Ginkgo biloba leaves (EGb 761) and its constituents. The effects described mostly in animals include those on cerebral blood flow, neurotransmitter systems, cellular redox state and nitric oxide level. Furthermore, we discuss the current status of clinical trials as well as undesired side effects of EGb 761.Received 21 November 2002; received after revision 8 March 2003; accepted 17 March 2003     

Konstante Stromwirkung und « funktionelle Polarität »

Summary Scheminzky found that the effect produced by passing a constant current through a frog depended on the direction of the current. While a descending current (directed from heat to foot) produced paralysis, an ascending current produced convulsions.Scheminzky believed that it was impossible to explain this phenomenon in any other way than by a functional polarity, involving a special microstructure in the cells of the spinal cord.—We found that the observed effect is in no way connected with the direction of the current, and that, on the contrary, it depends exclusively upon the electrical charge applied to the upper centres. The cathode on the spinal cord produces convulsions while the anode does not. The effects remain unchanged if the second electrode is removed from the spinal cord and applied to some spot outside the central nervous system. If, following extirpation of a small segment of the spinal cord, one electrode is placed on the head and the second electrode is applied to the cut surface of the lower part of the spinal cord, a perfect reversal of theScheminzky phenomenon can be obtained.TheScheminzky phenomenon is thus reduced to the well-known stimulating effect of the cathode and the paralysing effect of the anode. The question why a current passing in one direction (descending) should produce the anodic effect, while a current passing in the opposite direction (ascending) should produce the cathodic effect, can easily be explained by another well-known fact, i.e. the dependance of the lower spinal centres upon the higher ones. If the brain and upper centres are nearer to the anode (descending current), their anelectrotonic elimination has a paralysing effect, while their catelectrotonic excitation with an ascending current produces convulsions.     

nDsbD: a redox interaction hub in the Escherichia coli periplasm

DsbD is a redox-active protein of the inner Escherichia coli membrane possessing an N-terminal (nDsbD) and a C-terminal (cDsbD) periplasmic domain. nDsbD interacts with four different redox proteins involved in the periplasmic disulfide isomerization and in the cytochrome c maturation systems. We review here the studies that led to the structural characterization of all soluble DsbD domains involved and, most importantly, of trapped disulfide intermediate complexes of nDsbD with three of its four redox partners. These results revealed the structural features enabling nDsbD, a ‘redox hub’ with an immunoglobulin-like fold, to interact efficiently with its different thioredoxin-like partners. Received 3 February 2006; received after revision 1 March 2006; accepted 5 April 2006     

Stem cells from human apical papilla decrease neuro-inflammation and stimulate oligodendrocyte progenitor differentiation via activin-A secretion

Secondary damage following spinal cord injury leads to non-reversible lesions and hampering of the reparative process. The local production of pro-inflammatory cytokines such as TNF-α can exacerbate these events. Oligodendrocyte death also occurs, followed by progressive demyelination leading to significant tissue degeneration. Dental stem cells from human apical papilla (SCAP) can be easily obtained at the removal of an adult immature tooth. This offers a minimally invasive approach to re-use this tissue as a source of stem cells, as compared to biopsying neural tissue from a patient with a spinal cord injury. We assessed the potential of SCAP to exert neuroprotective effects by investigating two possible modes of action: modulation of neuro-inflammation and oligodendrocyte progenitor cell (OPC) differentiation. SCAP were co-cultured with LPS-activated microglia, LPS-activated rat spinal cord organotypic sections (SCOS), and LPS-activated co-cultures of SCOS and spinal cord adult OPC. We showed for the first time that SCAP can induce a reduction of TNF-α expression and secretion in inflamed spinal cord tissues and can stimulate OPC differentiation via activin-A secretion. This work underlines the potential therapeutic benefits of SCAP for spinal cord injury repair.     

De l'emploi d'isotopes radioactifs artificiels,dans le but d'exercer un effet radio-biologique localisé

Summary One of the authors has previously reported on a method which consists in the utilization of an artifical radioactive isotope (Zn⁶³), suspended in a suitably prepared solution ofpectin, for the production oflocalized biological radiation effects.This « macromolecular occlusion » of the radioactive isotope enables one to perform intraperitoneal injections (in cases of cancer of the ovaries with severe metastatic peritoneal extension), evidently also instillations in cavernous organs, and furthermore direct intratumoral injections, without diffusion of the radioactivity outside the treated areas, as shown both by autoradiographs and controls of blood and urine specimens with a Geiger counter.The authors investigated further whether this procedure would also be suitable for obtaining, by means ofintravenous injections, alocalized radiation effect within thelungs, as presumably the radiozinc, held in the large molecules of pectin, could thus be retained in the pulmonary capillaries. Intravenous injections of such a pectin solution containing radiozinc were performed on rabbits, and autoradiographic controls gave evidence of this expected fixation within the lungs.For the purpose of preliminary clinical investigation 40 millicuries of Zn⁶³ suspended in 6 cm³ of a 3 p. c. isotonic pectin solution were injectedintravenously in a female patient with mainly pulmonary metastases of a previously operated hypernephroma. This patient had been also submitted to X-ray therapy. In spite of a poor general condition, the injection was well tolerated. Autoradiographic controls showed quite clearly that the radioactivity remains precisely localized within the pulmonary areas. No radioactivity whatsoever was demonstrated with the counter in the urine eliminated by this patient after the injection, a fact which points to a rather amazing accuracy of the fixation of the radiozinc in the lungs. This first clinical experience seems quite interesting in view of improving the therapeutic possibilities of pathological, especially neoplastic pulmonary conditions.     

The effect of shock on blood oxidation-reduction potential

Oxidation-reduction (redox) potential measurements were made in the blood of rabbits subjected to hemorrhagic shock followed by treatment with a mild oxidizing agent (albumin). Control redox potential reading corrected for pH was –8.8±1.3 millivolts (mV) in arterial blood (A) and –18.0±2.0 mV in venous blood (V). This A-V difference indicated that hydrogen equivalents coming from muscle and other tissues were partially consumed in the lungs. A 20-mV drop on the V and a 13 mV on the A side was seen after shock. This did not fully return to control 2 h after return of the shed blood. Infusion of 2 g of albumin/kg/h raised the V redox potential to control, but it returned to untreated levels when the albumin was discontinued. The reductive load imposed on the animal by shock appeared to be large and not readily reversed by reperfusion or by the quantity of albumin given. Thus, it may be concluded that cellular respiration had not been adequately restored. This reductive load may impede recovery by suppression of cellular respiration and other cell and organ functions.     

Tissue protection against oxidative stress

We used an enhanced luminescence technique to study the response of rat tissues, such as liver, heart, muscle and blood, to oxidative stress and to determine their antioxidant capacity. As previously found for liver homogenate, the intensity of light emission (E) of tissue homogenates and blood samples, stressed with sodium perborate, is dependent on concentration, and the dose-response curves can be described by the equation E=a·C/exp(b·C). Theb value depends on the antioxidant defence capability of the tissues. In fact, it increases when homogenates are supplemented with an antioxidant, and is correlated with tissue antioxidant capacity, evaluated by two previously set up methods both using the same luminescence technique. Our results indicate that the order of antioxidant capacity of the tissues is liver>blood>heart>muscle. Thea value depends on the systems catalysing the production of radical species. In fact, it is related to the tissue level of hemoproteins, which are known to act as catalysts in radical production from hydroperoxides. The equation proposed to describe the dose-response relation is simple to handle and permits an immediate connection with the two characteristics of the systems analysed which determine their response to the pro-oxidant treatment. However, the equation which best describes the above relation for all the tissues is the following: E=·C/exp(·C). The parameter assumes values smaller than 1, which seem to depend on relative amounts of tissue hemoproteins and antioxidants. The extension of the analysis to mitochondria shows that they respond to oxidative stress in a way analogous to the tissues, and that the adherence of the dose-response curve to the course predicted from the equation E=a·C/exp(b·C) is again dependent on hemoprotein content.     

Postnatal variations of extracellular free calcium levels in the rat. Influence of undernutrition

Summary Ontogenetic changes in calcium activity were directly measured using an ion-selective micropipette in rat blood plasma and olfactory bulb extracellular fluid. Significant differences were observed according to the age and the nutritional state of the animal.We are grateful to Prof. W. Simon, Swiss Federal Institue of Technology, for providing the neutral carrier Ca²⁺ ion exchanger.     

In vivo and in vitro differences between leukocytic uptake of oligodeoxynucleotides

Development and application of therapeutic oligonucleotides rely on proper analysis of binding and uptake. We have used several model oligodeoxynucleotides (ODNs) to analyze binding/uptake by rat and human leukocytes. Here we describe: (1) differences between in vivo and in vitro uptake of ODNs to rat leukocytes, (2) differences after injection of lipopolysaccharide (LPS), (3) large in vitro differences between primary mononuclear cells in PBS, plasma and blood, and (4) differences of ODN uptake between rat and human leukocytes. Our data show that ODN uptake by primary blood cells was different in PBS, plasma and blood. In addition, LPS treatment increased ODN uptake by leukocytes in blood, indicating that pathological conditions may influence ODN uptake. Furthermore, ODN uptake in rat and human blood is also different, suggesting that preclinical ODN uptake data from rat blood cannot easily be extrapolated to the human condition. Received 17 December 2007; received after revision 16 January 2008; accepted 5 February 2008     

Intertidal habitat: does the shore level affect the nutritional condition of the shanny (Lipophrys pholis,Teleostei, Blenniidae)?

The nutritional condition of the shanny (Lipophrys pholis, Teleostei, Blenniidae), an intertidal fish, is affected by the shore level on whihc it dwells. Depending on the altitude within the intertidal, zone access to food is restricted for a certain time. A progressive decrease in the feeding time for an individual remaining on an increasingly higher shore level leads to a poorer nutritional condition compared to an individual staying at a lower shore level. Trade-off mechanisms between the feeding time and competition for space and/or predation pressure seem to be responsible for the still high abundance ofL. pholis on the upper shore. Possible consequences for growth and reproduction as well as distribution patterns are discussed.     

Antioxidant survey to assess antagonism to redox stress using a prokaryotic and an eukaryotic system

Using a prokaryote (Escherichia coli) and a metazoa-resembling eukaryote (Ochromonas danica), we surveyed antioxidants which might overcome redox stress imposed by menadione sodium bisulphite (MD) and buthionine sulphoximine (BSO). BSO oxidant stress was evident only inO. danica; MD oxidant stress was evident in both organisms. Glutathione, its precursors, e.g. cysteine, homocysteine, and 2-oxo-4-thiazolidine carboxylic acid, and red blood cells, emerged as prime antioxidants for relieving BSO and MD oxidant stress. BSO and MD oxidant activity and antioxidant-annulling effect inO. danica were judged comparable to those found in animal cells whereas the resultsE. coli were not entirely equivalent. TheO. danica system emerged as a practical, rapid, and useful system for pinpointing oxidant stressors and antioxidants, and shows promise for studies with mammalian systems.     

Spontaneous morphine withdrawal from the rat spinal cord

Summary A characteristic and reproducible sign of narcotic withdrawal is the naloxone induced increase in arterial pressure. In morphine-dependent rats allowed to undergo spontaneous withdrawal (6–24 h) and then transected at the spinal C-1 level, arterial pressure was maintained at a significantly higher level than either spinal-transected nondependent controls or morphine-dependent, spinal-transected rats pithed from C-1 to L-4. These findings indicate that the morphine-dependent spinal cord, independent of supraspinal influences, is able to exhibit an autonomic component of spontaneous withdrawal.This study was supported by the Medical Research Service of the Veterans Administration. A preliminary report of aspects of this work appeared in Soc. Neurosci. Abs.10 (1984) 1113.     

The effect of shock on blood oxidation-reduction potential.

Oxidation-reduction (redox) potential measurements were made in the blood of rabbits subjected to hemorrhagic shock followed by treatment with a mild oxidizing agent (albumin). Control redox potential reading corrected for pH was -8.8 +/- 1.3 millivolts (mV) in arterial blood (A) and -18.0 +/- 2.0 mV in venous blood (V). This A-V difference indicated that hydrogen equivalents coming from muscle and other tissues were partially consumed in the lungs. A 20-mV drop on the V and a 13 mV on the A side was seen after shock. This did not fully return to control 2 h after return of the shed blood. Infusion of 2 g of albumin/kg/h raised the V redox potential to control, but it returned to untreated levels when the albumin was discontinued. The reductive load imposed on the animal by shock appeared to be large and not readily reversed by reperfusion or by the quantity of albumin given. Thus, it may be concluded that cellular respiration had not been adequately restored. This reductive load may impede recovery by suppression of cellular respiration and other cell and organ functions.     

Defensive use of an acquired substance (carminic acid) by predaceous insect larvae

Larvae of two insects, a coccinellid beetle (Hyperaspis trifurcata) and a chamaemyiid fly (Leucopis sp.), feed on cochineal insects and appropriate their prey's defensive chemical, carminic acid, for protective purposes of their own.H. trifurcata discharges the chemical with droplets of blood (hemolymph) that it emits when disturbed;Leucopis sp. ejects the compound with rectal fluid. Ants are thwarted by these defenses, which are compared with the previously-described defense of a pyralid caterpillar (Laetilia coccidivora) that disgorges carminic acid-laden crop fluid. The defensive fluid of all three larvae contains carminic acid at concentrations spanning a range (0.2–6.2%) proven deterrent to ants. Many insects are known to appropriate defensive substances from plants. Insects that acquire defensive chemicals from animal sources may be relatively rare.Paper No. 124 in the series Defense Mechanisms of Arthropods; No. 123 is Epstein et al., J. Lepid. Soc. (in press).