The effect of chronic heat stress on cortisol levels in the antarctic fishPagothenia borchgrevinki

Radioimmunoassay was used to determine levels of the stress-inducible glucocorticoid, cortisol, circulating in the plasma of the extremely stenothermal Antarctic fishPagothenia borchgrevinki at rest and after heat stress. Fish sampled immediately after capture (–1.9°C) had low cortisol levels (10.4±1.4 ng ml^–1, mean±SEM) as did fish which were laboratory rested for 3 days. Sudden exposure to 5°C (48h) resulted in a peak cortisol value after 3 h (69.9±6.8 ng ml^–1) whereas exposure to 8°C (6h) resulted in a peak value after 1 h (73.5±8.0 ng ml^–1). At both temperatures levels remained significantly elevated (p<0.05) for the entire period of exposure. Increased temperature also resulted in a significant change in haemoglobin, haematocrit and mean cell haemoglobin concentration (MCHC) (p<0.05). Plasma lactate was significantly elevated only after exposure to 8°C (p<0.05). Plasma cortisol levels fromP. borchgrevinki are reported here for the first time and show this cryopelagic Antarctic species to have an unusual hormonal stress profile.     

Antarctic fishes survive exposure to carbon monoxide

The extensive in vivo conversion of haemoglobin to the carbon monoxide derivative has no discernible effect on the survival of the red-blooded Antarctic fishPagothenia bernacchii. Analysis of caudally sampled blood of cannulated specimens revealed that reconversion of carbon-monoxy haemoglobin to oxyhaemoglobin was complete within 48 hours. Thus, under stress-free conditions, haemoglobin is not necessary for survival ofP. bernacchii. Red-blooded Antarctic fishes can carry oxygen necessary for routine delivery dissolved in plasma, in a similar way to the haemoglobinless Channichthyidae, although they lack the morphological and physiological adaptations which allow the latter to prosper without any haemoglobin.     

Limitations on visual food-location in the planktivorous antarctic fishPagothenia borchgrevinki

Summary The visual threshold for food-location in the antarctic fishPagothenia borchgrevinki, is compared with light conditions and food availability under the ice. Even under the most favourable conditions for vision,P. borchgrevinki is operating close to its visual threshold and must often depend on non-visual mechanisms of prey detection.We thank J. D. Cargill and H. S. B. Weix for technical assistance, and the staff of Scott Base for their support. Funding was provided by the New Zealand University Grants Committee, and logistic support by the Antarctic Division of the Department of Scientific and Industrial Research.     

Occurrence of adrenaline and noradrenaline cells in the adrenal gland of the gentoo penguin (Pygoscelis papua)

Resumen Se investigó con microscopía óptica y electrónica y aplicando la técnica de glutaraldehido-plata la distribución y características de las células que contienen adrenalina y noradrenalina en el pinguino Papúa (Pygosceles papua).

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I thank Dr. J. H.Tramezzani and Dr. G. F.Wassermann for their important advice. I thank also Mrs.Isabel López de Farías for her technical assistance. This work was supported in part by the Argentine Antarctic Institute.     

Temperature dependence of neurotransmitter release in the antarctic fishPagothenia borchgrevinki

Summary The quantal contents of endplate potentials from extraocular muscles of an antarctic fishPagothenia borchgrevinki were measured over a range of temperatures. Quantal release was maximal at about 5°C but showed little dependence on temperature between –2°C and 10°C. Above 10°C quantal content declined until release ceased about 18°C. In view of the fact that the ambient temperature at which these fish live is constant at –1.9°C, the results suggest thatPagothenia borchgrevinki is only partially adapted to its environment despite 25 million years acclimatization.The authors wish to thank the staff of New Zealand's Scott Base, Antarctica, without whose cooperation the work would have been impossible. Mr J. Quinn of the Auckland University Physiology Department gave invaluable assistance with computing and electronics and the Antarctic Division of the New Zealand Department of Scientific and Industrial Research provided transportation and logistic support. The project was financially supported by the New Zealand University Grants Committee and the Auckland University Research Committee. S.P. was a Senior Fellow of the N.Z. Medical Research Council.     

Extraordinary levels of cadmium and zinc in a marine sponge,Tedania charcoti Topsent: inorganic chemical defense agents

The Antarctic marine spongeTedania charcoti has been shown to contain extraordinarily high natural concentrations of cadmium and zinc, which have in turn been correlated to the ability of the crude ethanol extract to modulate protein phosphorylation in chicken forebrain and to inhibit the growth of several test bacteria.     

Tubulin chaperone E binds microtubules and proteasomes and protects against misfolded protein stress

Mutation of tubulin chaperone E (TBCE) underlies hypoparathyroidism, retardation, and dysmorphism (HRD) syndrome with defective microtubule (MT) cytoskeleton. TBCE/yeast Pac2 comprises CAP-Gly, LRR (leucine-rich region), and UbL (ubiquitin-like) domains. TBCE folds α-tubulin and promotes α/β dimerization. We show that Pac2 functions in MT dynamics: the CAP-Gly domain binds α-tubulin and MTs, and functions in suppression of benomyl sensitivity of pac2Δ mutants. Pac2 binds proteasomes: the LRR binds Rpn1, and the UbL binds Rpn10; the latter interaction mediates Pac2 turnover. The UbL also binds the Skp1-Cdc53-F-box (SCF) ubiquitin ligase complex; these competing interactions for the UbL may impact on MT dynamics. pac2Δ mutants are sensitive to misfolded protein stress. This is suppressed by ectopic PAC2 with both the CAP-Gly and UbL domains being essential. We propose a novel role for Pac2 in the misfolded protein stress response based on its ability to interact with both the MT cytoskeleton and the proteasomes.     

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     

Beta-carotene affects gene expression in lungs of male and female <Emphasis Type="Italic">Bcmo1</Emphasis><Superscript>−/−</Superscript> mice in opposite directions

Molecular mechanisms triggered by high dietary beta-carotene (BC) intake in lung are largely unknown. We performed microarray gene expression analysis on lung tissue of BC supplemented beta-carotene 15,15′-monooxygenase 1 knockout (Bcmo1 ^−/−) mice, which are—like humans—able to accumulate BC. Our main observation was that the genes were regulated in an opposite direction in male and female Bcmo1 ^−/− mice by BC. The steroid biosynthetic pathway was overrepresented in BC-supplemented male Bcmo1 ^−/− mice. Testosterone levels were higher after BC supplementation only in Bcmo1 ^−/− mice, which had, unlike wild-type (Bcmo1 ^+/+) mice, large variations. We hypothesize that BC possibly affects hormone synthesis or metabolism. Since sex hormones influence lung cancer risk, these data might contribute to an explanation for the previously found increased lung cancer risk after BC supplementation (ATBC and CARET studies). Moreover, effects of BC may depend on the presence of frequent human BCMO1 polymorphisms, since these effects were not found in wild-type mice.     

Estimating and forecasting the long‐memory parameter in the presence of periodicity

We consider one parametric and five semiparametric approaches to estimate D in SARFIMA (0, D, 0)_s processes, that is, when the process is a fractionally integrated ARMA model with seasonality s. We also consider h‐step‐ahead forecasting for these processes. We present the proof of some features of this model and also a study based on a Monte Carlo simulation for different sample sizes and different seasonal periods. We compare the different estimation procedures analyzing the bias, the mean squared error values, and the confidence intervals for the estimators. We also consider three different methods to choose the total number of regressors in the regression analysis for the semiparametric class of estimation procedures. We apply the methodology to the Nile River flow monthly data, and also to a simulated seasonal fractionally integrated time series. Copyright © 2007 John Wiley & Sons, Ltd.     

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.     

Effect of NPV of the armywormMythimna (Pseudaletia) separata on the silkwormBombyx mori

Summary Nuclear polyhedrosis virus (NPV) material of the armywormMythimna (Pseudaletia) separata was administered in the form of PIBs and free virus rods to the larvae of silkwormBombyx mori. Routes used for administration were topical, intrahaemocoelic and oral. The larvae were treated with following concentrations: 10×10⁵ PIBs/L, 10×10⁶ PIBs/L, 10×10⁷ PIBs/L, 10×10⁸ PIBs/L. In all the 3 experiments, the larvae showed neither any signs and symptoms, nor mortality due to polyhedrosis. Thus it appears that the NPV ofM. (P.) separata is safe for the silkwormB. mori.Acknowledgments. We are grateful to the Karnatak University, Dharwad, for awarding UGC Junior Fellowship to S.G.D. and for financial assistance made available to S.B.M. We thank Professor M. Appaswamy Rao, Department of Zoology, for providing the necessary facilities. Our thanks are also due to Mr B. Chavannawar for supplying mulberry leaves and to the Government Silk Farm, Hindalaga, Belgaum, for providing silkworm eggs.     

Insulin activates Gαil,2 protein in rat hepatoma (HTC) cell membranes

Insulin action is initiated by binding to its cognate receptor, which then triggers multiple cellular responses by activating different signaling pathways. There is evidence that insulin receptor signaling may involve G protein activation in different target cells. We have studied the activation of G proteins in rat hepatoma (HTC) cells. We found that insulin stimulated binding of guanosine 5′-O-(3-thiotriphosphate) (GTP-γ-³⁵S) to plasma membrane proteins of HTC cells, in a dose-dependent manner. This effect was completely blocked by pertussis toxin treatment of the membranes, suggesting the involvement of G proteins of the Gα _i/Gα _o family. The expression of these Gα proteins was checked by Western blotting. Next, we used blocking antibodies to sort out the specific Gα protein activated by insulin stimulation. Anti-Gα _il,2 antibodies completely prevented insulin-stimulated GTP binding, whereas anti-Gα _o,i3 did not modify this effect of insulin on GTP binding. Moreover, we found physical association of the insulin receptor with Gα _i1,2 by copurification studies. These results further support the involvement of a pertussis toxin-sensitive G protein in insulin receptor signaling and provides some evidence of specific association and activation of Gα _i1,2 protein by insulin. These findings suggest that Gα _i1,2 proteins might be involved in insulin action. Received 23 September 1998; received after revision 23 November 1998; accepted 25 November 1998     

Abnormal organization of ovarian follicles in the mutantl(1)su(f) mad-ts ofDrosophila melanogaster

Summary The morphology of ovarian follicles in the temperature-sensitive female-sterile mutantl(1)su(f) ^mad-ts has been studied by means of light and electron microscopy. As the follicle cells gradually degenerate at the restrictive temperature (29°C) the follicles become increasingly disorganized with respect to shape and polarity.Acknowledgments. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 46). We wish to thank Mrs R. Koppa for excellent technical assistance and Mrs G. Mahlke for photographic work. We are also indebted to Dr D. Zissler, Dr E. Gateff and Prof. K. Sander for many helpful suggestions.     

Mid-anaphase arrest in S. cerevisiae cells eliminated for the function of Cin8 and dynein

S. cerevisiae anaphase spindle elongation is accomplished by the overlapping function of dynein and the kinesin-5 motor proteins, Cin8 and Kip1. Cin8 and dynein are synthetically lethal, yet the arrest phenotypes of cells eliminated for their function had not been identified. We found that at a non-permissive temperature, dyn1Δ cells that carry a temperature-sensitive cin8 – 3 mutation arrest at mid-anaphase with a unique phenotype, which we named TAN (two microtubule asters in one nucleus). These cells enter anaphase, but fail to proceed through the slow phase of anaphase B. At a permissive temperature, dyn1Δ, cin8 – 3 or dyn1Δcin8 – 3 cells exhibit perturbed spindle midzone morphologies, with dyn1Δcin8 – 3 anaphase spindles also being profoundly bent and nonrigid. Sorbitol, which has been suggested to stabilize microtubules, corrects these defects and suppresses the TAN phenotype. We conclude that dynein and Cin8 cooperate in anaphase midzone organization and influence microtubule dynamics, thus enabling progression through the slow phase of anaphase B. Received 10 August 2008; received after revision 22 October 2008; accepted 27 October 2008     

Hemolymph proteins in an ascitic condition,induced by lethal1(3)gl tumorous tissue inDrosophila hydei

Summary Drosophila hydei larvae homozygous for the gene1(3)gl may survive to the age of 3 weeks, become bloated and be incapable of metamorphosis. Wild-type flies after the injection of a fragment of the1(3)gl disc, which has been previously culture in vivo for 40–500 days, also become bloated (ascitic reaction). In the hemolymph of both the advanced1(3)gl larvae and ascitic adult hosts the accumulation of a specific protein is observed.Acknowledgments. This work was supported by the Swiss National Science Foundation under Grant No.3.792-0.76 and the Georges und Antoine Claraz-Schenkung. We thank Prof. H. Gloor, University of Geneva, and Prof. P.S. Chen, University of Zürich, for advice and criticism.     

Influence of cyclic nucleotides on protein synthesis in vascular smooth muscle

Summary The incorporation of leucice-¹⁴C into protein in bovine mesenteric arteries was augmented by cyclic GMP (10^–3 M) and decreased by cyclic AMP (10^–3 M). There was no effect of 5 AMP (10^–3 M). The phosphodiesterase inhibiting drugs theophylline (10^–3 M) and papaverine (5×10^–5 g/ml) both decreased the leucine-¹⁴C incorporation.We are indebted to Mrs.Lena Burlin for hear assistance. Finacial support has been provided by the Swedish State Medical Research Council (No. 04X-101X-4498).     

A role for the hinge/ear domain of the β chains in the incorporation of AP complexes into clathrin-coated pits and coated vesicles

The clathrin-associated adaptor protein (AP) complexes drive the polymerization of clathrin in coated pits to form coated vesicles. It has previously been shown that the carboxyl-terminal hinge/ear domain of the β2 chain contains a binding site for clathrin and that removal of this domain from APs or from isolated β2 chains abrogates their ability to form clathrin coats in vitro. We show here that the hinge/ear domain is necessary for efficient incorporation of AP complexes into coated pits and coated vesicles in cells, a result that is consistent with the view that the β chains indeed provide an important interaction between the AP complexes and clathrin. Received 7 April 1997; received after revision 22 May 1997; accepted 28 May 1997     

Characterization of DNAs fromCoprinus lagopus andMucor azygospora

Resumé Les rapports G+C des ADN deCoprinus lagopus etMucor azygospora ont été étudiés. Le profil de fusion indique que l'ADN duC. lagopus est composé de deux fractions, une principale (90%) de rapport G+C 52 moles pourcent, une mineure (10%) de G+C 32 moles pourcent. Par contre l'ADN deM. azygospora contient une fraction unique de G+C 38 moles pourcent. L'étude de la cinétique de réassociation DNA:DNA montre que la dimension génomique («genome size») deC. lagopus est de 2×10¹² et qu'il y a moin de 10% de DNA à séquences répétées de nucléotides.

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This research was supported in part by the U.S. Atomic Energy Commission Contract No. AT (40-1) 4182 and the Research Corporation, New York, to S.K.D. We are grateful to ProfessorG. Turian, University of Geneva, for making possibleM. Ojha's participation in this research.