Application of intracellular ATP determination in lymphocytes for HLA-typing

The amount of adenosine triphosphate (ATP) in human lymphocytes was determined using a technique based on light emission from a bioluminescent reaction with luciferin-luciferase. The amount of ATP changed when cells were incubated in the presence of specific HLA antisera and complement. For determination of intracellular ATP a modified method was applied, which was based on reduction of extracellular ATP by the addition of ATPase. The results of titration of an anti-human lymphocyte serum using the bioluminescence assay were in agreement with the results of fluorescence vitality staining. Bioluminescent HLA-determination in 57 cell samples each tested with 5 different antisera also gave good agreement (95.8%) with the conventional method. From these experimental data the calculated ATP content per lymphocyte was 0.135 ± 0.058 pg ATP.     

Osmotic responses of preimplantation mouse and bovine embryos and their cryobiological implications

Cells subjected to the events occurring before, during, and after freezing and thawing are exposed to major changes in the osmotic pressure of the surrounding medium; i.e., the osmolalities can exceed 30. An important question in understanding the mechanisms of injury is whether cells respond as ideal osmometers to these strongly anisotonic solutions. Mouse and bovine embryos from eight-cell to blastocyst stage were used to investigate the question. They were found to behave as ideal osmometers at room temperature over a wide range of tonicities; i.e., from four times isotonic to almost 1/3 times isotonic, ideality being defined by a Boyle-van't Hoff equation. Embryo volumes increased from 40 to 200% of isotonic over this range and survivals of mouse embryos were unaffected. However, outside this range the membrane apparently becomes leaky and the survival of mouse embryos drops sharply. Osmolalities rise to high values during freezing and the paper develops the thermodynamic equations to show how computed cell volumes as a function of subzero temperature can be translated into the Boyle-van't Hoff format of cell volume as a function of the reciprocal of osmolality.     

Vitamin K-dependent carboxylase: the carboxylation of exogenous substrates in different systems

Two types of solid-phase carboxylase, SPC-II and SPC-X, have been prepared from the livers of warfarin-treated cows. Their enzymatic activities were compared with substrate-free carboxylase in microsomes from normal cows and substrate-bound carboxylase in microsomes from warfarin-treated cows. A number of exogenous substrates for carboxylase have been purified and tested. We found that large substrates, such as descarboxyprothrombin, are carboxylated only by substrate-free carboxylase and not by the substrate-bound enzyme. No differences in apparent Km values between solid-phase carboxylases II and X were observed.     

Hybrid dysgenesis-induced Y chromosomal sterility in Drosophila melanogaster

Summary Dysgenic hybrids of Drosophila melanogaster were screened for the induction of mutations in the Y chromosomal fertility genes. Out of 2,417 Y chromosomes analysed 13 male steriles (ms (Y)) were isolated. This high rate of mutation is most probably due to the unusually large physical size of the fertility genes.     

Zur anatomie der ammoniten: Tintenbeutel,kiemen, augen

The ink sacs within the living chambers of ammonites as hitherto published seem to be remains of the stomach-crop-complex. Ammonites had no ink sacs. Fossilised remains of gills were found within the living chambers of three ammonoid specimens (generaOphiceras, Hildoceras, Eleganticeras). Their state of preservation is not good enough to identify their numbers. Their shape seems to be not quite dissimilar to that of recentNautilus. Reflections concerning the eyes of ammonites suggest that they were of the pinhole type like those of recentNautilus.     

Effects of fertilization and liming on the chemical soil conditions and element distribution in forest soils

Summary The distribution and storage of major elements in acid soils from a spruce and a beech forest was investigated after fertilization of NH₄NO₃ and KCl followed by Ca and Mg fertilization by 2 liming applications. All fertilizers were applied on top of the soil without mixing. Most of the added Ca and Mg was detected in the humus layer, a significant part of it still in carbonatic form. The effect of liming on mineral soil pH is very low, and was only observed in the 0–10 cm layer. However, base saturation of the mineral soil increased. The storage of C and N of the humus layer was not affected. N fertilization increased the N storage of the soil only under beech, but was followed by heavy NO₃-losses with seepage water under spruce. High leaching rates for K were also found in the spruce stand. The amount of K that was not leached increased the pool of exchangeable K in the deeper soil layer.     

Fluorescence methods for localizing proteinases and proteinase inhibitors in skeletal muscle

Summary Proteinases and proteinase inhibitors have become suspect in a wide variety of muscle wasting conditions that might be treatable if knowledge of the cellular locale and function of these molecules were known. Fluorescent probes have been useful in the localization of proteinases in muscle samples from human and animal specimens. These include the histochemical localization of proteinases based on the specific fluorescence of hydrolysis product derivatives, but this approach has been limited to the lysosomal proteinases because of the acidic requirements of the trapping reaction of the primary reaction product. Immunohistochemical techniques do not have the same restrictions and a number of lysosomal and nonlysosomal proteinases have been identified in muscle by this means. Unfortunately, they do not yield any information as to the activity of the enzymes. This is an important consideration since the extracellular environment contains a number of proteinase inhibitors, some of which may be internalized by the cell.     

Determination of the hydraulic conductivity of Lamprothamnium by use of the pressure clamp

By use of the pressure-clamp technique, the hydraulic conductivity of the brackish-water alga Lamprothamnium was found to be 5·10^-6 cm s^-1 bar^-1. The dimensions of the internodes are so small that it is possible, for the first time, to measure a complete volume relaxation upon clamping the turgor pressure to a preset value by a feedback control of the pressure probe. As theoretically predicted, the values of the hydraulic conductivity obtained from the initial slope of the volume relaxation induced by the pressure clamp are in agreement (within experimental error) with those obtained from the half-time of the relaxation process. The cell volume also obtained from the analysis of the volume relaxation is the osmotically effective cell volume and is therefore slightly smaller than the value obtained by taking the dimensions of the cell including the cell wall.Abbreviations and symbols Lp hydraulic conductivity - P turgor pressure - S_v initial slope of volume relaxion - T_1/2 half-time of volume relaxation Dedicated to Professor Dr. H. Ziegler on the occasion of his 60th birthday     

Isolation and characterization of a new thermophilic and autotrophic methane producing bacterium:Methanobacterium thermoaggregans spec. nov.

Methanobacterium thermoaggregans is a new thermophilic autotrophic rod-shaped methane producing bacterium. The organism likes to form aggregates during growth and utilizes only H₂ and CO₂ as substrates. Growth optimum is at 65°C with a doubling time of 3.5 h. Optimal growth occurs at pH-values between 7 and 7.5. The addition of yeast extract to the mineral salt medium stimulates growth. The DNA base composition is 42 mol% G+C. The organism was isolated from mud taken from a cattle pasture. Because of its optimal growth temperature and its tendency to form aggregates the nameMethanobacterium thermoaggregans is suggested.Abbreviations G+C Guanine+cytosine