Extracellular enzyme activity: Indications for high short-term variability in a coastal marine ecosystem

Extracellular -glucosidase, -glucosidase, and aminopeptidase activity variations (measured by use of fluorogenic substrate analogs) at a coastal station in the Mediterranean Sea were investigated over a 1-year period. A 27-h cycle and daily measurements were made in a summer situation. We observed strong relative diurnal variations, compared to seasonal variations, in - and -glucosidase. Within 24 h, 0–100% of both - and -glucosidase were found in the dissolved phase. The aminopeptidase activities did not show a strong diurnal variation, but day to day variations were similar in magnitude to seasonal changes. Consistently, high proportions of all three enzymes were found in the dissolved phase on a seasonal scale. Seasonal measurements at 50- and 100-m depths showed a weak negative dependency on depth for extracellular enzyme activity. The potential importance of both hourly and daily changes in extracellular enzyme activity and of free enzymes is considered. Correspondence to: M. Karner     

A fluorescent modification of the Sakaguchi reaction on arginine

Summary The reactionproduct of arginine with 2,4-dichloro--naphthol proved to be fluorescent. This fluorescence could be used for the histochemical localization of arginine. The modification described resulted in a long lasting yellow-orange fluorescence of various arginine-containing tissues.Thanks are due to Mr. H. van Kooten for making the photograph.     

Selective inhibition of prostaglandin biosynthesis by gold salts and phenylbutazone

Gold salts and phenylbutazone selectively inhibit the synthesis of PGF_2α and PGE₂ respectively. Lowered production of one prostaglandin species is accompanied by an increased production of the other. Selective inhibition by these drugs was observed in the presence of adrenaline, reduced glutathione and copper sulphate under conditions when most anti-inflammatory compounds inhibited PGE₂ and PGF_2α syntheses equally. It is postulated that selective inhibitors may have a different mode of action in vivo and beneficial effects may be related to the endogenous ratio of PGE to PGF required for normal function.     

Sphingosine-1-phosphate effects on PKC isoform expression in human osteoblastic cells.

Sphingosine-1-phosphate (S1P) has been shown to participate in the proliferative process in human osteoblasts.(1) The mitogenic effect of S1P has been postulated to involve two signaling pathways, the Gi linked protein receptor pathway and the PKC pathway. To define the possible role of PKC isoforms in osteoblastic cell proliferation, the effects of S1P on PKC isoform expression was determined. While PKC lambda was minimally detected, the isoforms alpha, delta and iota were all found to be highly expressed by the human osteoblast. In human osteoblastic cells, S1P induced a 25% increase in the expression of PKC alpha and approximately a 30% increase in the expression of PKC iota. S1P did not have an effect on PKC delta expression. Pretreatment with pertussis toxin (PT) led to an inhibition of the observed S1P effects on the expression of the alpha and iota isoforms.     

The relative importance of sediment and water column supplies of nutrients to the growth and tissue nutrient content of the green macroalga Enteromorpha intestinalis along an estuarine resource gradient

Large blooms of opportunistic green macroalgae such as Enteromorpha intestinalis are of ecological concern in estuaries worldwide. Macroalgae derive their nutrients from the water column but estuarine sediments may also be an important nutrient source. We hypothesized that the importance of these nutrient sources to E. intestinalis varies along a nutrient-resource gradient within an estuary. We tested this in experimental units constructed with water and sediments collected from 3 sites in Upper Newport Bay estuary, California, US, that varied greatly in water column nutrient concentrations. For each site there were three treatments: sediments + water; sediments + water + Enteromorpha intestinalis (algae); inert sand + water + algae. Water in units was exchanged weekly simulating low turnover characteristic of poorly flushed estuaries. The importance of the water column versus sediments as a source of nutrients to E. intestinalis varied with the magnitude of the different sources. When initial water column levels of dissolved inorganic nitrogen (DIN) and soluble reactive phosphorus (SRP) were low, estuarine sediments increased E. intestinalis growth and tissue nutrient content. In experimental units from sites where initial water column DIN was high, there was no effect of estuarine sediments on E. intestinalis growth or tissue N content. Salinity, however, was low in these units and may have inhibited growth. E. intestinalis growth and tissue P content were highest in units from the site with highest initial sediment nutrient content. Water column DIN was depleted each week of the experiment. Thus, the water column was a primary source of nutrients to the algae when water column nutrient supply was high, and the sediments supplemented nutrient supply to the algae when water column nutrient sources were low. Depletion of water column DIN in sediment + water units indicated that the sediments may have acted as a nutrient sink in the absence of macroalgae. Our data provide direct experimental evidence that macroalgae utilize and ecologically benefit from nutrients stored in estuarine sediments.     

A conserved interaction with the chromophore of fluorescent proteins

The chromophore of fluorescent proteins, including the green fluorescent protein (GFP), contains a highly conjugated imidazolidinone ring. In many fluorescent proteins, the carbonyl group of the imidazolidinone ring engages in a hydrogen bond with the side chain of an arginine residue. Prior studies have indicated that such an electrophilic carbonyl group in a protein often accepts electron density from a main-chain oxygen. A survey of high-resolution structures of fluorescent proteins indicates that electron lone pairs of a main-chain oxygen-Thr62 in GFP-donate electron density into an antibonding orbital of the imidazolidinone carbonyl group. This n→π* electron delocalization prevents structural distortion during chromophore excitation that could otherwise lead to fluorescence quenching. In addition, this interaction is present in on-pathway intermediates leading to the chromophore, and thus could direct its biogenesis. Accordingly, this n→π* interaction merits inclusion in computational and photophysical analyses of the chromophore, and in speculations about the molecular evolution of fluorescent proteins.     

Can We Disrupt the Sensing of Honey Bees by the Bee Parasite Varroa destructor?

BackgroundThe ectoparasitic mite, Varroa destructor, is considered to be one of the most significant threats to apiculture around the world. Chemical cues are known to play a significant role in the host-finding behavior of Varroa. The mites distinguish between bees from different task groups, and prefer nurses over foragers. We examined the possibility of disrupting the Varroa – honey bee interaction by targeting the mite''s olfactory system. In particular, we examined the effect of volatile compounds, ethers of cis 5-(2′-hydroxyethyl) cyclopent-2-en-1-ol or of dihydroquinone, resorcinol or catechol. We tested the effect of these compounds on the Varroa chemosensory organ by electrophysiology and on behavior in a choice bioassay. The electrophysiological studies were conducted on the isolated foreleg. In the behavioral bioassay, the mite''s preference between a nurse and a forager bee was evaluated.ConclusionsThese data indicate the potential of the selected compounds to disrupt the Varroa - honey bee associations, thus opening new avenues for Varroa control.     

A fluctuating salinity regime mitigates the negative effects of reduced salinity on the estuarine macroalga, Enteromorpha intestinalis (L.) link

We tested the response of Enteromorpha intestinalis to fluctuating reduced salinity regimes which may occur in coastal estuaries due to both natural and anthropogenic influences. In a fully crossed two factor experiment, we subjected E. intestinalis to 0, 5, 15 and 25 psu water enriched with nutrients for 1-, 5-, 11- and 23-day periods. Each period was followed by 24 h of exposure to 25 psu (ambient) water that was not nutrient enriched. Following 24 h in ambient salinity water, algae were returned to reduced salinity conditions for the appropriate period and the cycle continued over the 24 days for which all treatments were maintained. Exposure to 0 psu for 5 days or longer resulted in loss of pigmentation, decreased wet and dry biomass, increased wet wt:dry wt ratios, decreased removal of nitrogen (N) and phosphorus (P) from the water column and an accumulation of NH(4) in the water column. More frequent exposure to ambient salinity in the 1-day treatment mitigated these effects. Across all salinity levels tested, biomass increased as frequency of exposure to ambient salinity increased. At all durations of exposure to low salinity tested, biomass increased as salinity level increased. We conclude that growth of E. intestinalis is decreased by reduced salinity. E. intestinalis is able to withstand exposure to 0 psu but there is a temporal limit to this tolerance that is somewhere between 1 and 5 days. Populations of E. intestinalis in coastal estuaries may suffer from freshwater inputs if salinity conditions are persistently reduced.     

Sublethal damage during cryopreservation of rainbow trout sperm

Although cellular damage during cryopreservation of freshwater fish spermatozoa has been reported in several studies, there is a lack of correlation between this damage and the fertility rates of eggs using postthawed milt. The apparent lack of such correlation may be due to other undetected sublethal cryodamage, which could affect the cell functionality and viability. This may be extremely important for freshwater fish spermatozoa whose ability to fertilize the egg requires dilution in water or hypoosmotic solutions, an hazardous environment for the cells. This study tested the change in cell permeability during cryopreservation, using Hoechst 33258 to assess cell permeability. The permeability of spermatozoa at different times after dilution in several hypoosmotic media were investigated. In the first trial, fresh semen, sperm diluted in freezing media (CPT), and freeze/thawed semen were studied. Three CPT were tested (Me2SO, DMA, and methanol). In the second trial, the addition of egg yolk as a membrane stabilizer was investigated. Samples were frozen at -20 degreesC/min in a programmable cooler and thawed in a 25 degreesC water bath. Dilution in the CPTs slightly increased the susceptibility of cells to damage but freezing/thawing caused a dramatic increase in the fragility of cells, which were killed in a few seconds after their contact with the hypoosmotic solutions. Egg yolk provided a significant protection to the membrane, allowing the cells a greater and more prolonged survival in the fertilization media. Samples frozen with Me2SO displayed the best results. These results are consistent with the achieved fertility rates that demonstrated sublethal cryodamage in the function of the sperm membrane that was not detected by standard procedures. Copyright 1998 Academic Press.