Differential permeabilization of membranes by saponin treatment of isolated rat hepatocytes. Release of secretory proteins.

Monolayer cultures of rat hepatocytes were treated with increasing concentrations of saponin (prepared from Gypsophila plants) for 30 min at 6 degrees C. Differential permeabilization of the intracellular membranes could be demonstrated: at 0.040 mg of saponin/ml the plasma membrane was permeabilized, as assessed by the release of 50% of the total cellular amount of lactate dehydrogenase, and at 0.20 mg/ml the endoplasmic reticulum was permeabilized, as measured by the release of 50% of pulse-35S-labelled albumin. The Golgi complex was permeabilized at an intermediate saponin concentration, as indicated by the release of homogeneously 35S-labelled albumin; about half the intracellular albumin is located in this organelle. At 1.0 up to 5.0 mg of saponin/ml 90-95% of the radioactively labelled albumin was released. Even at 5.0 mg/ml less than 10% of the membrane of the endoplasmic reticulum was solubilized, as judged by the degree of release of a membrane-bound enzyme specific for this organelle. These results demonstrate the usefulness of saponin as a tool for investigating the interior of different intracellular compartments.     

Fish cleaning behaviour of shrimp

Cleaning behaviour of five species of shrimp from three families was studied at three different geographic locations in an effort to gain a quantitative understanding of cleaning behaviour, and to compare a broad cross-section of cleaner shrimp species. Two shrimp from the genus Periclimenes , two from the genus Lysmata , and one from the genus Stenopus were used and 27 hours of recorded laboratory observations were made for each of the five shrimp species.
All shrimp species were inactive most of the observed time, and most spent less than 2% of the observed time cleaning fish hosts. Also, the shrimp spent more time cleaning the ventral rather than the dorsal surface of the fish because they were reluctant to board the fish. However, evenness in cleaning does not appear to be an indicator of overall excellence in cleaning because the two best cleaners (based on number and duration of cleaning bouts) were among the least even in their cleaning.
The fish cleaning behaviour of the shrimp appeared to be strictly stereotypic in form, but the stimulus-response and the total amount of cleaning differed greatly among the five species. A Cleaning Efficiency Index (CEI) was created in an attempt to incorporate significant aspects of the cleaning behaviour. According to this CEI, Lysmata grabhami was by far the most efficient (best) cleaner, CEI = 55–51, compared to the others; Stenopus hispidus , 33–78; Periclimenes pedersoni , 6–29; Periclimenes yucatunicus , 5–60; and Lysmata californien , 2–12.
The cleaners most widely distributed geographically have the highest CEI scores, while the most localized cleaners have the lowest CEIs. This relationship may allow the CEI score to be useful in determining a cleaner shrimp's potential geographical distribution, and may also serve as an indicator for the degree of phylogenetic relationship to other cleaner shrimps.     

Lysogenic Conversion in Bacillus amyloliquefaciens H Affecting Viral Adsorption

Lysogenization of Bacillus amyloliquefaciens H with bacteriophage PK renders the bacteria resistant to superinfection with bacteriophage of PK type.     

Linkage of a gene for macular corneal dystrophy to chromosome 16.

Autosomal recessive macular corneal dystrophy (MCD) is a heterogeneous disorder leading to visual impairment. Sixteen American and Icelandic families (11 type I and 5 type II) were analyzed for linkage, by use of 208 polymorphic microsatellite markers. A significant maximum LOD score Zmax of 7.82 at a maximum recombination fraction (thetamax) of .06 was found with the 16q22 locus D16S518 for MCD type I. In addition, a peak LOD score of 2.50 at a recombination fraction of .00 was obtained for the MCD type II families, by use of the identical marker. These findings raise the possibility that MCD type II may be due to the same genetic locus that is involved in MCD type I.     

Seed transmission of turnip yellow mosaic virus in winter turnip and winter oilseed rapes

This is the first record of seed transmission of turnip yellow mosaic virus (TYMV) in oilseed and turnip rapes. The seed transmission of TYMV in a naturally infected winter turnip rape (Brassica napus var. silvestris) cultivar Perko PVH was investigated. By ELISA 1.6%, 3.2% and 8.3% seed transmission of the virus was found in seed of plants from three localities. The proportion of infected seeds produced by artificially infected plants of winter oilseed rape (Brassica napus ssp. oleifera) and winter turnip rape cultivars was determined. The virus transmission rate, expressed as the proportion of virus-infected plants which germinated from the seed was for the oilseed rape cvs Jet Neuf 0.1%, Solida 0.4%, Silesia 0.8%, Darmor 1.2%, SL-507 0.2%, SL-509 0.0% and for the winter turnip rape cv. Perko 1.5%. ELISA cannot be used in direct tests on bulk seed lots to estimate proportion of infected seed, but must be used on germinated seedlings.     

Characterization of a folding intermediate of human carbonic anhydrase II: probing local mobility by electron paramagnetic resonance.

The spin-labeling method was used to investigate human carbonic anhydrase, HCA II, undergoing unfolding induced by guanidine-HCI (Gu-HCI). The spin-probe, N-(2,2,5,5-tetramethyl-1-yloxypyrrolidinyl-3-yl)iodoacetamide, was attached covalently to the single cysteine (position 206) in the enzyme. The electron paramagnetic resonance spectrum of the folded structure showed the characteristic slow motional spectra. When the concentration of the denaturing agent, Gu-HCI, was gradually increased, new spectral components with narrower lines evolved to give complex electron paramagnetic resonance spectra, apparently containing superimposed contributions from several components of different mobility. By a differentiation technique, it was possible to follow the relative increase of the narrow components as a function of Gu-HCI concentration. The amplitude of difference spectra versus Gu-HCI concentration showed two distinct maxima, indicating the existence of a folding intermediate state/structure. The results were found to agree with optical absorption data, which showed similar transitions at the same Gu-HCI concentrations. From line-shape simulations assuming a Brownian diffusion model, the rotational diffusion constants for the spin-label in the folded, folding intermediate, and unfolded structures were determined. The relative abundances of the three conformations in the region 0-4 M Gu-HCI were obtained by least squares fitting of the simulated spectra to the experimental ones. The folding intermediate was found to have a maximum population of 39 +/- 4% at approximately 0.7 M Gu-HCI.     

Aspects of hadrosaurian cranial anatomy

Supraorbital bones in Saurolophus angustirostris are described, and their presence in all hadrosaurs is suggested. Frontal-nasal and premaxillar-nasal fontanellae are distinguished in hadrosaurs; their presence is explained as connected with growth and considered to he responsible for the variability of crest structures. New data indicating the presence of a cartilaginous diverticulum nasi within the circumnarial depression in Saurobphus ongustirostris are presented. A physiological (respiratory and/or thermoregulatory) function of the nasal diverticulum is proposed.     

Nutrient content and dynamics in north Swedish shrub tundra areas

Forbs and leaves of deciduous shrubs had high concentrations of Ca, Mg, K, P, and N. Deciduous dwarf shrubs had intermediate concentrations but higher than evergreen dwarf shrubs. Monocots, cryptogams, woody and belowground tissues had low concentrations.
Plant nutrient concentrations and nutrient content in soil organic matter increased from dry towards moist tundra areas.
The residence time of nutrients was considerably less than ten years in surface litter, but several decades or centuries in total organic matter. The longest residence time was found in the moist part of the tundra.
N, K, and P in Betula nana leaves were translocated to a great extent prior to leaf fall, whereas Mg and particularly Ca were only slightly translocated. As on other tundra areas shortage of nutrients probably limits plant growth. In that case short supply of N and F seems most probable due to retention in litter and soil organic matter.     

Structural and functional differences between carbonic anhydrase isoenzymes I and II as studied by site-directed mutagenesis

Site-specific mutagenesis has been used to replace amino acid residues in the active site of human carbonic anhydrase II with residues characterizing carbonic anhydrases I. Previous studies of [Thr200----His]isoenzyme II [Behravan, G., Jonsson, B.-H. & Lindskog, S. (1990) Eur. J. Biochem. 190, 351-357] showed that His200 is important for the specific catalytic properties of isoenzymes I. In this paper some properties of two single mutants, Asn62----Val and Asn67----His, as well as a double mutant, Asn67----His/Thr200----His, are described. The results show that neither Val62 nor His67 give rise to isoenzyme-I-like properties, while the double mutant behaves like the single mutant with His200. At pH 8.9, the variant with Val62 has a higher value of kcat/Km for CO2 hydration than unmodified isoenzyme II, whereas the variant with His67 has an enhanced kcat value. The replacement of Asn62 with Val results in a 20% increase of the 4-nitrophenyl acetate hydrolase activity. For the double mutant, the esterase activity is quite close to that calculated on the assumption that the effects of the two single mutations on the free energy of activation are additive.