Comparative electrophoretic studies of crustacean neurosecretory hyperglycemic and melanophore-stimulating hormones from isolated sinus glands

Summary By disc electrophoresis of sinus gland extracts fromOrconectes limosus, Pacifastacus leniusculus andCarcinus maenas, marked differences in the electrophoretic mobility were demonstrated for the hyperglycemic neurohormone (HGH) from the 3 species. In the astacuran materials, 2 peaks of activity were observed whereas inCarcinus material only 1 peak could be demonstrated with certainty. Melanophore-stimulating activity (MDH) was resolved into several peaks, but 2 predominant peaks with almost identical mobility were observed in all 3 species. Electrophoresis of sinus gland material from the above species and fromHomarus gammarus, Procambarus clarkii, Astacus leptodactylus, A. astacus, Uca pugilator, Cancer magister, Eriocheir sinensis andHemigrapsus oregonensis revealed that all astacuran HGHs fall in one group of substances with low mobility whereas some of the brachyuran hormones (fromCarcinus, Uca andCancer) moved faster. The hormones ofEriocheir andHemigrapsus have mobilities similar to those of the astacuran hormones, but, like the other brachyuran hormones, they are not hyperglycemic in Astacura. HGH was found to be associated with the strongest staining band in the pherograms from all species investigated. It amounts to at least 10% of total sinus gland protein, that is about 0.5 g/gland inOrconectes and 0.6 g/gland inCarcinus. Accumulation of neurosecretory material in the neurohaemal organ is demonstrated by comparison of sinus gland and medulla terminalis extracts. In SDS gels, a large fraction of material (containing the HGH) in the mass range of 6000–8000 daltons is found in the the sinus gland whereas this fraction is barely demonstrable in the medulla terminalis. It was investigated whether cysteine-rich carrier proteins comparable to the vertebrate neurophysins exist in the sinus gland. No positive results were obtained either by aldehyde fuchsin staining of gels or by analysis of³⁵S-labeled sinus gland material. noreactivity in gels after electrophoresis of sinus gland extracts. No positive reaction was obtained with eluates from HGH containing slices. Recent amino acid analyses of HGH (Kleinholz, 1975; Keller and Wunderer, in preparation) do not suggest any similarities between crustacean HGH and vertebrate glucagon.     

Energieabhängige 63Ni-Aufnahme bei Alcaligenes eutrophus Stamm H 1 und H 16

Zusammenfassung Die Kinetik der Aufnahme von ⁶³Ni wurde an zwei Stämmen von Alcaligenes eutrophus untersucht, die Nickelionen für das chemolithotrophe Wachstum benötigen. Mit Kohlendioxid als einziger Kohlenstoffquelle wird das Wachstum durch niedrige Konzentrationen von Nickel gefördert, wobei das Optimum der Wachstumsförderung bei 0,3 M Nickel lag. Höhere Nickelkonzentrationen wirkten hemmend. Das heterotrophe Wachstum mit Fructose wurde durch Nickelionen nicht gefördert. — Übertragen in Phosphatpuffer, der von Schwermetallionen befreit worden war, zeigten autotroph gewachsene Zellen eine rasche Aufnahme von ⁶³Ni, sofern Wasserstoff, Sauerstoff und Kohlendioxid zugegen waren. Dabei wurde Nickel innerhalb von 60 min aus dem umgebenden Medium bis zur 280 fachen Konzentration in den Zellen angehäuft. Die beobachtete Ni-Aufnahme zeigte ein Temperaturoptimum bei etwa 29° C und wurde durch Hemmstoffe wie Arsenit, Jodacetat, Methylenblau, Natriumazid und Natriumcyanid stark beeinträchtigt. Andere Schwermetallionen (Zn, Co, Mn und Cu) verminderten die Nickelaufnahme nur geringfügig. Durch ⁵⁸NiCl₂ und Toluol wurde der Efflux von ⁶³Ni aus den Zellen gefördert. Die Beobachtungen lassen darauf schließen, daß Nickelionen durch einen energieabhängigen Prozeß in chemolithotroph gewachsenen Zellen dieser Stämme angehäuft werden.

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Energy-dependent ⁶³Ni-uptake by Alcaligenes eutrophus strains H 1 and H 16

Kinetic studies of the uptake of ⁶³Ni were undertaken with two strains of Alcaligenes eutrophus, known to require nickel ions for chemolithotrophic growth. Using carbon dioxide as sole carbon source, growth is stimulated by low concentrations of nickel with optimum concentration for growth stimulation at about 0.3 M nickel. Higher nickel concentrations were inhibitory. Heterotrophic growth on fructose was not stimulated by nickel ions.-Upon transfer into phosphate buffer freed of heavy metal ions, autotrophically grown cells exhibited rapid uptake of ⁶³Ni which was dependent upon th presence of hydrogen, oxygen and carbon dioxide. Within 60 min nickel was accumulated from the medium, reaching 280-fold concentration in the cells. The observed uptake exhibited a temperature optimum at about 29° C and was markedly inhibited by metabolic inhibitors such as arsenite, iodoacetate, methylene-blue, sodium azide and sodium cyanide. Other heavy metal ions (Zn, Co, Mn and Cu) only slightly inhibited ⁶³Ni-uptake. The efflux of ⁶³Ni from the cells was stimulated by ⁵⁸NiCl₂ and by toluene. These data indicate that nickel ions are accumulated by an energy dependent mechanism in chemolithotrophically grown cells of these strains.

     

Measurement of fast light-induced disc shrinkage within bovine rod outer segments by means of a light-scattering transient

A fast light-induced light-scattering transient, previously found in rod outer segment suspension, the so-called P-signal (Hofmann, K.P., Uhl, R., Hoffmann, W. and Kreutz, W. (1976) Biophys. Struct. Mechanism 2, 61–77), is described in more detail.The effect has the same action spectrum as rhodopsin bleaching. It is not regenerated with 11-cis retinal.The response is not linear with light-intensity for flashes which bleach more than 2.0% of rhodopsin; it saturates at an intensity corresponding to 15% rhodopsin bleaching.The wavelength- and scattering angle dependence lead to the conclusion that the change in light-scattering reflects a shrinkage of an osmotic compartment of the rod outer segment.The only compartment which we found to be intact in our rod outer segment preparations was the disc or rod sac; therefore, the effect must be attributed to a light-induced shrinkage of the rhodopsin-containing disc organelles.The overall effect (15% of rhodopsin is bleached) is in the range of 0.5–1.5% of the original volume.A light-induced passive cation-efflux from the disc, e.g. of Ca²⁺, can be ruled out as a possible molecular origin of the disc-shrinkage in our preparations.     

Manipulation of Whole-Lake Ecosystems and Long-Term Limnological Observations in the Brandenburg — Mecklenburg Lake District,Germany

An accompanying technical use of ecological effects from pelagic food-web interactions (biofiltration) or from sediment (nutrient remobilization) can speed up recovery of lake ecosystems and can make restoration programs more effective after external load reductions. At present the comprehensive use of multiple ecotechnological controls for water quality is limited because our knowledge of the many complex interactions, indirect effects and above all the long-term effects of such operations is insufficient. Beginning in the 1960s we started long-term limnological analyses of the trophic interactions between community structure and the flow of matter under “manipulated” conditions in whole-lake experiments at Lake Stechlin (manipulated water circulation and waste heat), Lake Haussee (biomanipulation) and Lake Fuchskuhle (lake-dividing). This paper summarises changes in abiotic and biotic structures and functions of the manipulated lake ecosystems and discusses the possibilities and limitations of “whole-lake” experiments.     

Low environmental selenium availability as an additional determinant for goiter in East Java, Indonesia?

Iodine deficiency, which is most visibly indicated by goiter, is highly prevalent in Indonesia. Since 1994, Indonesia has a decree that all salt used for human, livestock, and industry must be iodized. However, despite the increased distribution of iodized salt, pockets with significantly higher prevalence of goiter still remain. This situation may be consequence of selenium (Se) deficiency. This study aimed to assess the Se level in the environment of goiter prevalent areas. Five hundred eleven school children participated in this study. Goiter was measured using both ultrasound and palpation. Ninety-nine eggs were collected from free-living chicken in 11 villages, and the Se contents of egg yolk and egg white were determined by neutron activation analysis. In the villages studied, Se concentration in egg yolk ranged from 0.15 to 1.52 Μg/g and in egg white from 0.18 to 2.97 Μg/g. The prevalence of goiter measured by palpation ranged from 18.4% to 70% and by ultrasound from 0% to 100%. Because of the inconsistency of goiter rate measured by palpation and ultrasonography, the question remains whether low availability of Se in the environment might be an additional contributing factor for goiter.     

A new method for induction of specific polyclonal antibodies using immunostained proteins on nitrocellulose membrane, and HIV 1 core antigen and Escherichia coli verotoxin as examples

Abstract Two polyclonal antibodies against verotoxin 1 and the core protein p24 of HIV 1 were raised in mice by a new immunization procedure. Both proteins were transferred to nitrocellulose, reacted with polyspecific antisera and the antigen-antibody complexes were then visualized by immunostaining. For preparation of antisera the stained protein bands were cut from the nitrocellulose sheets and implanted subcutaneously into the backs of BALB/c mice, without any adjuvant. A single booster was given 4 weeks later by implanting a second strip. All mice produced high titers of antibody directed against the antigen used for immunization. Thus, antibodies of high specificity can be elicited against protein bands in stained immunoblots.     

Phase-plane analyses of the Michaelis-Menten reaction equations

The irreversible Michaelis-Menten scheme may be reduced to a pair of autonomous first-order differential equations. The phase-plane behaviour of these is investigated.     

Factors affecting the level and localization of the transferrin receptor in Trypanosoma brucei

Transfer of bloodstream-form Trypanosoma brucei variant 221a from calf serum to dog serum-based medium induces acute iron starvation, as the transferrin receptor (Tf-R) of variant 221a binds dog Tf poorly. We show here that transfer to dog serum induces a 3-5-fold increase in Tf-R mRNA and protein within one doubling time (8 h). Because iron stores are still high 8 h after transfer, we infer that the signal for Tf-R overproduction is the decreased availability of cytosolic iron when cellular iron import drops. Up to 30% of the extra Tf-R spills out of the flagellar pocket onto the pellicular surface. Because the 5-fold increase in Tf-R is accompanied by a 5-fold increase in bovine Tf uptake, the up-regulation of Tf-R levels in response to Tf starvation helps the trypanosome to compete for limiting amounts of Tf. We noted that Tf-R levels also vary in calf serum medium. Cells in dense cultures contain up to 5-fold more Tf-R mRNA and protein than in dilute cultures. Only one-tenth of the extra Tf-R reaches the pellicular surface. The increase cannot be explained by a lack of Tf or to cell density sensing but is due to pericellular hypoxia. Our results show that bloodstream-form trypanosomes can regulate the expression of the two Tf-R subunit genes and the localization of their gene products in a flexible manner. This flexibility is made possible by the promoter-proximal position of the two genes in the variant surface glycoprotein expression site.