Thermische Denaturierung von Kaltblüter-Enzymproteinen

Summary The temperature, which leads to 50% reduction of catalytic activity by heat denaturation, has been determined for 8 different enzymes from cod muscle, as being in the range between 30 and 52°C. Therefore, there are no indications of a generally different heat resistance of enzymes from cold-blooded animals as compared with those from warm-blooded animals. The same conclusion is derived from calculations ofQ ₁₀-values, measured between +37 and – 37°C for cathepsin and glycylglycine dipeptidase.

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Die Untersuchungen wurden dankenswerterweise durch eine Beihilfe des Bundeswirtschaftsministeriums Bonn unterstützt. HerrnDegener, i. Fa. Hanseatische Hochseefischerei A.G., Bremerhaven-F., verdanken wir das Untersuchungsmaterial.     

Über Struktur und Aktivität der den H2O2-Zerfall katalysierenden Cu2+-Komplexe. — VI. Differenzierung von nativer RNS und nativer DNS bzw. denaturierter DNS auf Grund der katalytischen Eigenschaften

Summary The catalysis of H₂O₂ decomposition by Cu²⁺-complexes of RNA and DNA has been investigated. It is shown that both complexes decompose H₂O₂, but only the Cu²⁺-RNA-system shows peroxidative activity too, e.g. only in this case the nucleotide bases are degraded. Thermal denaturation of DNA also leads to a Cu²⁺-complex with peroxidative activity, the latter being dependent on the degree of denaturation.

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V. Mitteilung:S. Petri, H. Sigel undH. Erlenmeyer, Helv. chim. Acta49, 1778 (1966).

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12. Mitteilung überMetallionen und H ₂ O ₂; 11. Mitteilung:H. Ch. Curtius, P. Anders, R. Zell, H. Sigel undH. Erlenmeyer, Helv. chim. Acta49, 2256 (1966).     

Dissociation-constants of metal-ion-complexes with alkaline phosphatase from pig kidney

Summary Using metal-ion buffers it was possible to remove Zn²⁺, Mg²⁺ and Mn²⁺ ions of pig kidney alkaline phosphatase reversibly. The dissociation constants obtained are K_EMg:4·10^–7 M, K_EMn:4·10^–8 M and K_EZn:8·10^–13 M (22°C, pH:9.6, :0.07).Acknowledgement: The authors thank Dr.H. U. Wolf for helpful suggestions and valuable discussion and MissH. Köth for technical assistance.     

Effects of Mn2+ and Mg2+ on the pigeon liver pyruvate kinase

Riassunto Mn²⁺ and Mg²⁺ attivano la piruvato cinasi di fegato di piccione in maniera distinta. In presenza di basse concentrationi di fosfoenolpiruvato Mn⁺ é piú efficace di Mg²⁺ ed é attivatore dell'enzima saturato da Mg²⁺. Piruvato cinasi (EC 2.7.1.40).

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This work was supported by a grant from the Consiglio Nazionale delle Ricerche, Roma, Italia.Silvia Baldi is a fellow of the Italian C.N.R.     

In vitro fertilization of mouse and hamster eggs after freezing and thawing

Summary Mouse eggs (12–14%) and hamster eggs (87–94%) appear normal upon thawing after having been stored for 30 min to 25 h at –70°C and –196°C. While 89–98% of the normal appearing hamster eggs are capable of fertilization in vitro, only 4–13% of the mouse eggs can be fertilized. The ability of fertilized eggs to develop in culture and their viability upon transfer to pseudopregnant recipients is under investigation.This work was supported by grants from the U.S. Public Health Service and the Ford Foundation. Sincere thanks are due to Prof.Y. Nishikawa of Kyoto University, Japan, for advice and to Mrs.Rose Bartke and Ms.D. M. Hunt for assistance.     

Die Wirkung von 4,6-Dinitro-o-kresol auf die Atmung eines experimentellen Rattentumors bei verschiedenen Temperaturen

Summary (1) The oxygen uptake of an experimental sarcoma of the rat was measured under the influence of 4,6-Dinitro-o-cresol in a range of temperature from 22.5 to 47.5°C.(2) Dinitrocresol inhibited the respiration of the tumor at low temperatures and stimulated it with rising temperatures, similar to its action on tissues of poikilothermic animals.(3) An explanation is given in terms of the theory ofJohnson and our own preceding results.     

Die Wirkung der Temperatur auf das Verhalten der Gewebsatmung poikilothermer und homoiothermer Tiere unter dem Einfluss von 4,6-Dinitro-o-kresol

Summary (1) The liver respirationin vitro of mice and toads was studied under the influence of 4,6-Dinitro-o-cresol at temperatures varying from 17,5 to 42,5°C.(2) The percentage activity of the oxygen uptake of the mouse liver decreased progressively under dinitrocresol with rising temperatures, whereas the respiration of the toad liver showed inhibition at low temperatures and activation at higher temperature.(3) The results were interpreted following the theory of inhibitory types ofJohnson.     

Effect of heat treatment on the ATPase activity of various sarcoplasmic reticulum preparations.

Ca2+-stimulated ATPase activity of sarcoplasmic reticulum (SR) preparations is activated after a short period of preincubation at temperatures between 40 and 45 degrees C, but for temperatures higher than 48 degrees C pronounced denaturation is observed. Heat denaturation is decreased if Mg2+ or K+ are present during heat treatment.     

Zinc iodide-osmium tetroxide reactive substances in the matrix of granulated vesicles in the rat pineal nerves

Resumen Se demuestra que la reserpina depleciona los componentes de las vesículas sinápticas de los nervios pineales de la rata revelables por la mezcla de yoduro de zinc-tetróxido de osmio a 4°C durante dos horas y a 20°C durante 15 h mientras que lap-CPA sólo depleciona los componentes revelables a 4°C durante 2 h.

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This work has been supported by grants from the Universidad Nacional de Buenos Aires, the Consejo Nacional de Investigaciones Cientificas y Técnicas, Argentina and the National Institutes of Health (Nas. 2 ROI NS 06953-07 NEUA), USA.

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We are grateful to MissM. López for her skillful technical assistance.     

Aufhebung der Thermothyrin-A-Sekretion der Schilddrüse durch Fütterung mit Methylthiouracil

Summary G. Mansfeld demonstrated that in the serum of overheated animals a substance (thermothyrine A) is present which, injected into normal animals, decreases O₂-consumption. Serum of thyroidectomized animals has no effect.Dogs and rabbits were treated daily with 0.10 g per kg methylthiouracil during 4 weeks, and were than subjected for 5 hours to a temperature of 34–35° C which raised their body temperature by 0.5–1.5° C. 2.5 cm³ of serum obtained at the end of the 5 hours period failed to reduce O₂-consumption of normal rats, while sera of untreated dogs and rabbits produced after similar exposure to high temperature a fall of O₂-consumption by 14–48%. It is therefore evident that methylthiouracil not only inhibits the formation of thyroxine but of thermothyrine A as well.The fact that thermothyrine A contains no iodine proves conclusively that the action of thiouracil compounds cannot be exclusively an inhibition of iodinization.     

Kälte als Auslöser einer Glykogen-Speicherung während der Oogenese vonMusca domestica

Summary InMusca domestica reared under standard conditions (21 °C), glycogen is stored in the oocyte towards the end of egg development. By 1–3 days exposure to lower temperatures (4 °C), a glycogen deposition can be released already in young follicles. This premature glycogen synthesis is not restricted to the ooplasm. Carbohydrates are also found in the nurse cells and the follicle epithelium. Similar results were formerly obtained by inhibition of oogenetic protein synthesis.

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Herrn Prof. Dr. Dr. h.c.Bernhard Rensch zum 70. Geburtstag gewidmet.     

Establishment of a cell line (XTC-2) from the South African clawed toad,Xenopus laevis

Résumé On décrit une méthode pour l'établissement d'une culture de cellules du crapaud a griffes sudafricain,Xenopus laevis, afin de l'infecter avec des arbovirus. Les cellules (XTC-2) ont subi une culture continue pendant 3 ans. Elles ont un nombre diploïde de chromosomes. Elles furent cultivées aux températures de 22°C et de 28°C dans un milieu de culture dilué de NCTC 109 avec 10% de sérum de veau foetal.

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We thank ProfessorD. S. Bertram, Director of the Department of Entomology at the School, for his continued interest; Mr.A. O. Langi for technical assistance and Dr.L. Hamilton, Middlesex Hospital Medical School, London, for supplying theXenopus tadpoles. We are especially grateful to the Wellcome Research Laboratories, Beckenham, for supplying medium NCTC 109, and to Mr.D. J. Richmond of the Laboratories for responding to our request for medium promptly and willingly. The work is supported in part by a grant from the Medical Research Council of Great Britain.     

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.     

Temperature acclimation of Mg2+Ca2+-myofibrillar ATPase from a cold-selective teleost,Salvelinus fontinalis: a compromise solution

Summary Brook trout (Salvelinus fontinalis, Mitchill) were acclimated over 15 weeks to either +4°C or +24°C. The effects of temperature on myofibrillar Mg²⁺Ca²⁺-ATPase activities were investigated. In contrast to goldfish, temperature acclimation does not alter the kinetic properties of the brook trout myofibrillar ATPase. Activation energy (G^#) is lower and substrate turnover number is higher than values previously reported for cold-adapted stenotherms. Properties of brook trout ATPase appear to be a compromise enabling function across a broad temperature range. The different strategies of adapting to seasonal temperature variations are briefly discussed.The authors are grateful to the Wellcome Trust for financial support.-Correspondence should be addressed to I.A.J.     

Is glucagon involved in cold acclimatization?

Summary Cold acclimatization in rats at 5°C for 2 weeks caused a significant elevation of plasma glucagon concentration, accompanied by increased plasma FFA and glucose levels. Acute cold exposure at 5°C for 5 or 60 min did not affect these parameters in plasma.We are grateful to Drs.H. Ohhara andA. Kihara, Sapporo Medical College, for help in setting up glucagon radioimmunoassay.     

A method for distinguishing Tetrodotoxin from Saxitoxin,by comparing their relative stabilities when heated in acid solution

Resumen La tetrodotoxina pierde toxicidad rapidamente cuando se calienta en solución acida por debajo de pH 2, mientras que la saxitoxina es estable. Se puede distinguir facilmente entre estos venenos calentándolos a 100°C en solution pH 1.0 durante 20–30 min.

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This work was carried out whileC. J. Waterfield was on industrial training leave from Bath University of Technology. We wish to thank MissJ. Tulloch for her assistance.     

Affinity chromatographic preparation of arterial heavy meromyosin subfragment-1

Summary Heavy meromyosin subfragment-1 (HMM S-1) was prepared by papain digestion of arterial myosin or actomyosin and was purified by agarose-ATP affinity chromatography. Proteolysis of crude arterial myosin suspensions was preceded by solubilization. HMM-S-1 thus obtained consisted mainly of a 90,000 dalton polypeptide and fully retained the K⁺- and Ca²⁺-ATPase of the parent myosin. Its affinity to agarose-ATP was comparable to that of skeletal muscle HMM S-1.The support of this work by an EMBO short time fellowship (to R. L.) and by the Deutsche Forschungsgemeinschaft (No. SFB 90, B 7) and the excellent technical assistance byM. Troschka andC. Köhler are gratefully acknowledged.     

Diffusion loading conditions determine recovery of protein synthesis in electroporated P3X63 Ag8 cells

Summary Using the suspension cell line P3X63 Ag8 we have studied the impact of the composition of the diffusion medium on cellular protein synthesis under standard electroporation conditions in TBS-Na. This buffer contains the high saline concentration usually present in electroporation-mediated DNA transfection. Electroporation in the presence of TBS-Na resulted in an immediate shut-off of protein synthesis, even though both FITC-dextran (M_r 40 kD) and Semliki Forest virus core protein (M_r 33 kD) were incorporated efficiently into the cytoplasm across the electropores at 0°C. Subsequent resealing of the pores was completed after a 5-min incubation at 37°C. When compared with control cells, overall protein synthesis of electroporated cells recovered slowly to resume a 30% activity after 1 h of incubation at 37°C. We have determined optimal conditions for diffusion loading (which necessitates the presence of ATP, GTP, amino acids, K⁺, Mg²⁺, and Ca²⁺) and resealing (in the presence of K⁺, Mg²⁺, and Ca²⁺), leading to a full and lasting recovery of protein synthesis within 5 min after pore closure.     

Membrane constants and sodium conductance of a single muscle fibre

Riassunto Nella singola fibra muscolare di cavia a 38°C sono state determinate, applicando la «square pulse analysis», la resistenza, la costante di tempo, la costante di spazio e la capacità della membrana a riposo. È stato inoltre possibile calcolare la massima conduttanza per il sodio,gn a, che è risultata di 35,3 mmho/cm².     

Über die Funktion der Lorenzinischen Ampullen der Selachier

Summary The afferent impulses from the ampullae ofLorenzini ofScyllium show, both in the intact animal and in the isolated preparation, a steady discharge at constant temperature with a frequency reaching a maximum at an average of 20° and decreasing continuously at higher and lower temperatures. The discharge stops between 5° and 30° on the average. Rapid cooling causes a temporary rise in frequency, while rapid warming causes a temporary drop in frequency. While the ampullae are not sensitive to a mechanical stimulus, they react definitely to a change in temperature of 0.05°C. The ampullae thus behave like the cold receptors of the homoiotherms.