35Cl nuclear magnetic resonance studies of anion binding to halophilic proteins: Ferredoxin from Halobacterium of the Dead Sea

The anion-binding characteristics of ferredoxin from Halobacterium of the Dead Sea have been studied by ³⁵Cl^? NMR. It is found that the binding constant of Cl^? to halophilic ferredoxin is ca. 0.09 at 28 °C and that the binding enthalpy is positive. It is also found that the correlation time for chloride ions bound to halophilic ferredoxin is about 10 ns. The effect on the ³⁵Cl^?1 NMR signal of adding competing anions is also studied. Halophilic proteins like ferredoxin which have a high negative charge bind anions with low affinity but the ³⁵Cl^? quadrupole relaxation technique can conveniently monitor such weak binding.     

Nearest-neighbor and next-nearest-neighbor effects in the proton NMR spectra of the oligoribonucleotides ApGpX and CpApX

The variable-temperature proton nmr spectra of the oligoribonucleotides in the series CpApX and the series ApGpX, X = A, G, C, U, together with the parent dimers CpA and ApG have been measured. A complete analysis of all the nonexchangeable base proton resonances and ribose H-1′ proton resonances was made. The presence of trends in the shielding abilities of the various bases at both the nearest-neighbor and next-nearest-neighbor positions were identified. The observed shieldings could be used to predict the chemical shifts of protons in related systems. Based on the empirical results from ribodinucleoside monophosphates, the temperature-dependent behavior of the J_1′2′ coupling constants of the triribonucleotides suggested that the compounds in the CpApX series stacked from the 5′-end to the 3′-end, while those in the ApGpX series stacked from the 3′-end to the 5′-end.     

Myosin light chain patterns of individual fast and slow-twitch fibres of rabbit muscles

Summary Single muscle fibres were isolated by microdissection from freeze-dried samples of rabbit psoas and soleus muscles. The individual fibres were typed according to qualitative histochemical reactions for succinate dehydrogenase or NADH-tetrazolium reductase and for alkaline Ca²⁺-activated myofibrillar myosin ATPase after acid or alkaline preincubation. Methods are described for electrophoretic analysis by means of polyacrylamide disc electrophoresis in the presence of SDS of total myofibrillar proteins in single fibres after pre-extraction of soluble proteins. Fast-twitch white fibres revealed a myosin light chain pattern characteristic of fast-type myosin with three light chains of apparent molecular weights of 22,300 (LC₁), 18,400 (LC₂) and 16,000 (LC₃). Fast-twitch red fibres were indistinguishable in this respect from fast-twitch white fibres and showed an identical pattern of myosin light chains. Slow-twitch fibres could be characterized by a myosin light chain pattern typical of myosin of slow-twitch muscles with peptides of the apparent molecular weights of 23,500 (LC_1Sa), 23,000 (LC_1Sb) and 18,500 (LS_2S). Slow-twitch fibres isolated from soleus as well as from psoas muscle were indistinguishable with regard to their myosin light chain patterns, thus suggesting that fibres of the same histochemical type correspond in their myosin light chain patterns irrespective of their origin from different muscles.Dedicated to the memory of Ernest Gutmann who has contributed so much to our knowledge on differentiation of muscle and who died on August 6, 1977     

Determination of the configurations of lactic and glyceric acids from human serum and urine by capillary gas—liquid chromatography

The separation of the enantiomers of lactic and glyceric acids can be achieved by capillary gas chromatography on SP-1000 using the corresponding O-acetylated menthyl esters. The structures of the derivatives were proved by proton magnetic resonance spectroscopy and mass spectrometry. The method has been used for the determination of the absolute configurations of lactic and glyceric acids isolated from serum and urine from different patients.     

Pulsation of nuclei in insect oocytes is reversibly inhibited by cytochalasin B

Oocyte nuclei of the dipteran insect Heteropeza pygmaea display swift pulsating movements during in vitro follicle formation in the ovaries. Low doses of cytochalasin B (CB) completely inhibit the nuclear movements within a few minutes and cause the nuclei to assume spherical shapes. If the drug is removed, nuclear pulsation is resumed within 5–10 min. Phalloidin and colchicine do not affect the nuclear movements. Actin is shown by indirect immunofluorescence microscopy to be present in considerable amounts all over the cytoplasm of the oocytes. It is concluded that microfilaments are responsible for pulsation of the oocyte nuclei, whereas microtubules are not involved.     

Protective Effects of Human Recombinant Mnsod in Adjuvant Arthritis and Bleomycin-Induced Lung Fibrosis

We have previously shown that human recombinant methionyl manganese superoxide dismutase (MnSOD) is more efficient than CuZnSOD as an anti-inflammatory agent in a model of acute inflammation (Carrageenan-induced pau edema). This effect was attributed to the prolonged half-life of MnSOD in blood (t_1/2 = 6 h vs. 10 min. respectively). In the present study, the two enzymes were compared in terms of their effectiveness in two systems: (I) Adjuvant-induced arthritis in rats, which is considered to be a model for the chronic situation of rheumatoid arthritis and (2) Bleomycin-induced lung fibrosis. which is a chronic situation believed to be mediated by oxygen free radicals.

Rats inflicted with adjuvant arthritis were treated during the period of maximal joint swelling (Days 15-21 after adjuvant injection) with MnSOD or CuZnSOD (12 to 50mg/kg, i.p. daily). MnSOD administration resulted in a 50-75% reduction of paw swelling, as well as inhibition of the elevation of serum globulins. A similar treatment with CuZnSOD gave merely marginal effects.

In the second system, lung fibrosis was induced in rats by intratracheal administration of bleomycin. MnSOD (50mg/kg, s.c.), administered 2 h before and then 2 and 4 days after bleomycin, markedly inhibited lung fibrosis, as evident from lung weight and collagen content measured by the 3rd week. By contrast, CuZnSOD administration did not give a significant effect. The results indicate that MnSOD is superior to CuZnSOD in the treatment of chronic inflammatory processes. In addition, they lend further support to the involvement of oxygen free radicals in bleomycin toxicity.     

Biotechnological production of unnatural L-amino acids from D,L-5-monosubstituted hydantions. II. L-α- and L-β-naphthylalanine

Summary Resting cells ofArthrobacter sp. (DSM 3745) with the ability to form L-tryptophan from D,L-5-(3-indolylmethy)hydantoin were used for the bioconversion of D,L-5-- and D,L-5--naphthylmethylhydantoin (D,L-5-- and D,L-5--NMH) to the corresponding L-amino acids. Under the optimal reaction conditions of pH 9.7 and 40°C specific productivities of 0.2 (-naphtylalanine) and 0.6 (-naphtylalanine) mM amino acid x g cell dry mass^–1 x h^–1 were obtained in a 0.1 M Na₂CO₃/NaHCO₃-buffer in a strirred bioreactor.     

Antibacterial and antimycotic effect of a newly discovered secretion from larvae of an endoparasitic insect,Pimpla turionellae L. (Hym.)

Three analogues of the peptidyl pheromone, pheromone of Saccharomyces kluyveri, synthesized based on the amino acid sequence proposed by Sato et al. (Agric Biol Chem 45:1531–1533, 1981) were tested for both shmoo-inducing and agglutinability-inducing actions. Purified natural pheromone of the yeast showed the highest activity among the peptides tested. When methionine in the peptides was oxidized, the activity decreased significatly. Pheromone of S. kluyveri induced sexual agglutinability in a cells of Saccharomyces cerevisiae, and shmoo in a cells of S. cerevisiae and S. kluyveri. a Pheromone of S. kluyveri had no agglutinability-inducing action on cells of S. cerevisiae. a Cells of S. kluyveri inactivated only pheromone of the same species, but a cells of S. cerevisiae inactivated pheromones of both S. cerevisiae and S. kluyveri.     

EGG ACTIVATION AND PARTHENOGENETIC REPRODUCTION IN INSECTS

1. Many insects reproduce by parthenogenesis. In one of the largest orders of the animal kingdom, the Hymenoptera, most of its members reproduce by arrhenotokous parthenogenesis. Egg activation in parthenogenetic animals obviously cannot be caused by fertilization of the egg. The question of what initiates egg development in parthenogenetically reproducing animals has been studied for a few insect species and is discussed in this article. 2. The grasshopper Melanoplus differentialis is one of several Orthoptera displaying accidental parthenogenesis. In this species, egg laying provides the stimulus to the completion of meiosis and start of embryonic development in unfertilized and probably also in fertilized eggs. The same holds true for the dipteran insect Drosophila melanogaster which exhibits rudimentary parthenogenesis, and for D. mercatorum showing accidental parthenogenesis. The precise way in which oviposition affects the egg is unknown. 3. The stick insect Carausius morosus reproduces by obligatory thelytoky. The triggering factor for removal of the meiotic block and initiation of embryonic development is oxygen from the air which penetrates to the egg through the micropyle immediately after oviposition. The oviposition act itself is not necessary for activation of the egg. 4. Comparative studies of the different types of oogenesis in the dipteran insect Heteropeza pygmaea show that in paedogenetically developing follicles meiotic arrest in prophase is of very short duration and a meiotic block at the end of oogenesis is absent. It is suggested that in this case triggering events for egg development are dispensable. On the other hand, under certain experimental conditions a meiotic block can be established in some of these follicles. 5. Investigations on the Ichneumonid wasp Pimpla turionellae have shown that unfertilized, male-determined eggs - and most likely also fertilized, femaledetermined eggs - are activated by mechanical stress exerted on the eggs during natural or imitated oviposition. This mechanical stress, in addition, activates a streaming system which is independent of meiotic completion and nuclear multiplication. Egg activation by egg distortion is also found in the Pteromalid species Nasonia vitripennis and occurs presumably in many other Hymenoptera. 6. Carausius morosus, Pimpla turionellae and Nasonia vitripennis are species with parthenogenetic reproduction for which the natural factors responsible for the initiation of egg development have been identified. The cases of Pimpla turionellae and Nasonia vitripennis are of particular interest because of the feasibility of artificially imitating the natural activating mechanism. 7. It is concluded that apart from fertilization various events at oviposition may trigger egg development. In addition, the occurrence of rudimentary parthenogenesis in many sexually reproducing animal species suggests that sperm entry and fertilization may frequently be necessary for the continuation of egg development rather than for its initiation.