Interrelationships between Rates of Microbial Production,Exopolymer Production,Microbial Biomass,and Sediment Stability in Biofilms of Intertidal Sediments

Abstract The upper few millimeters of intertidal sediment supports a varied biomass of microbial consortia and microphytobenthos. Many of these organisms release extracellular polymers into the surrounding sediment matrix that can result in sediment cohesion and the increased stability of the sediment. The relationship between the heterotrophic and autotrophic components of these biofilms is not well understood. A combination of mesocosm and field investigations were used to investigate the relationship between microbial production rate (algae and bacteria), the extracellular carbohydrates, biomass, and stability in conjunction with a variety of environmental factors. An inverse relationship was found between rates of algal production and sediment stability both in the field and in laboratory mesocosms, though the relationship was significant only in the field (P < 0.001). Stability of sediments increased with increasing bacterial production rate (P < 0.001). Positive correlations were found between sediment stability and a range of other variables, including algal biomass (P < 0.001), colloidal-S EPS (P < 0.001), colloidal-S carbohydrate (P < 0.01), colloidal-S EDTA (P < 0.01), and sediment water content (P < 0.001). Using the data acquired, a preliminary model was developed to predict changes in sediment stability. Chlorophyll a, water content, and colloidal-S EPS were found to be the most important predictors of stability in intact cores incubated under laboratory conditions. Differences observed in patterns of the surface (0–2 mm) distribution of colloidal-S carbohydrate and chlorophyll a when expressed on a dry weight or areal basis were attributed to effects of dewatering and concomitant changes in wet bulk density. The polymeric carbohydrate (colloidal-S EPS) component of the biofilms was not found to be a constant fraction of the colloidal-S carbohydrate extract, varying from 16 to 58%, and the percentage of polymer decreased logarithmically as chlorophyll a concentrations increased and the biofilms matured (P < 0.001). Changes in the relationships between these variables over the period of biofilm development and maturation highlight the difficulties in their use to predict sediment stability. Exopolymer concentrations were more closely correlated with algal biomass than with bacterial numbers. Rates of algal carbon fixation were considerably greater than those for bacteria, suggesting that the algae have a much greater potential for exopolymer production. It is suggested that the microphytobenthos secretions make a more important contribution to sediment stability. Received: 12 May 1999; Accepted: 13 October 1999; Online Publication: 24 March 2000     

Multicenter evaluation of the phosphorylcholine-coated biodivYsio stent in short de novo coronary lesions: The SOPHOS study

AIMS: The BiodivYsio trade mark stent (Biocompatibles Ltd, Farnham, UK) is coated with a phosphorylcholine (PC)-containing copolymer to confer biocompatibility. The SOPHOS (Study Of PHosphorylcholine coating On Stents) study was designed to assess the safety and efficacy of this novel coronary stent and by indirect comparison to indicate equivalence with other formal stent studies. METHODS AND RESULTS: Patients with angina and a single short (#x2A7F;12 mm) de novo lesion in a native coronary artery of >/=2.75 mm diameter were included. A total of 425 patients were allocated in 24 centers. Clinical data were collected at one-, six- and nine-month follow-up. Angiography was performed before and after the stent implantation. In addition, in the first 200 patients (SOPHOS A) angiography was routinely performed at six months. The following 225 patients (SOPHOS B) were merely followed up clinically. The primary end-point of the study, the six-month MACE-rate (MACE = Major Adverse Cardiac Events) was 13.4% (two cardiac death; five Q-wave/nine non-Q-wave myocardial infarctions (MI); nine CABG and 32 target lesion revascularization (TLR), which is similar to the calculated 15% MACE-rate in comparable reference studies. Secondary end-points included among others restenosis at six months in the SOPHOS A population. The target vessel diameter was 2.98 +/- 0.48 mm. Minimal lumen diameter pre/post procedure and at follow-up was 1.00 +/- 0.32, 2.69 +/- 0.37, 1.91 +/- 0.71 mm, respectively. The binary restenosis rate (>/=50% diameter stenosis at follow-up) was 17.7%. CONCLUSION: The coronary BiodivYsio stent is safe and effective as a primary device for the treatment of native coronary artery lesions in patients with stable or unstable angina pectoris. Clinical and angiographic results are in the statistical range of equivalence with comparable studies with other current stents.     

Serine 727 phosphorylation and activation of cytosolic phospholipase A2 by MNK1-related protein kinases

We have previously reported that in thrombin-stimulated human platelets, cytosolic phospholipase A(2) (cPLA2) is phosphorylated on Ser-505 by p38 protein kinase and on Ser-727 by an unknown kinase. Pharmacological inhibition of p38 leads to inhibition of cPLA2 phosphorylation at both Ser-505 and Ser-727 suggesting that the kinase responsible for phosphorylation on Ser-727 is activated in a p38-dependent pathway. By using Chinese hamster ovary, HeLa, and HEK293 cells stably transfected with wild type and phosphorylation site mutant forms of cPLA2, we show that phosphorylation of cPLA2 at both Ser-505 and Ser-727 and elevation of Ca(2+) leads to its activation in agonist-stimulated cells. The p38-activated protein kinases MNK1, MSK1, and PRAK1 phosphorylate cPLA2 in vitro uniquely on Ser-727 as shown by mass spectrometry. Furthermore, MNK1 and PRAK1, but not MSK1, is present in platelets and undergo modest activation in response to thrombin. Expression of a dominant negative form of MNK1 in HEK293 cells leads to significant inhibition of cPLA2-mediated arachidonate release. The results suggest that MNK1 or a closely related kinase is responsible for in vivo phosphorylation of cPLA2 on Ser-727.     

Comparative mapping of quantitative trait loci sculpting the curd of Brassica oleracea

The enlarged inflorescence (curd) of cauliflower and broccoli provide not only a popular vegetable for human consumption, but also a unique opportunity for scientists who seek to understand the genetic basis of plant growth and development. By the comparison of quantitative trait loci (QTL) maps constructed from three different F(2) populations, we identified a total of 86 QTL that control eight curd-related traits in Brassica oleracea. The 86 QTL may reflect allelic variation in as few as 67 different genetic loci and 54 ancestral genes. Although the locations of QTL affecting a trait occasionally corresponded between different populations or between different homeologous Brassica chromosomes, our data supported other molecular and morphological data in suggesting that the Brassica genus is rapidly evolving. Comparative data enabled us to identify a number of candidate genes from Arabidopsis that warrant further investigation to determine if some of them might account for Brassica QTL. The Arabidopsis/Brassica system is an important example of both the challenges and opportunities associated with extrapolation of genomic information from facile models to large-genome taxa including major crops.     

A laser-based method for measuring thermal nociception of cattle

We describe a method for measuring nociception in cattle using a CO(2) laser aimed at the caudal aspect of the metatarsi. In Experiment 1, infrared thermography showed that calves responded by lifting their legs when skin temperatures reached 45-55 degrees C. In Experiment 2a, the validity of the method was tested by comparing the response latencies of 14 calves to two power settings (2.25 W vs. 4.5 W) with each setting being applied six times. We found that both leg-lift latencies and tail-flick latencies were lower at the higher power setting, and the calves were more likely to respond by kicking than by simply moving the leg. The standard deviations between and within calves were smaller at the higher power setting, and the large within-calf variation means that at least three tests were required to obtain reliable measures that could differentiate between calves. In Experiment 2b, application of the laser at a range of power settings (2.0, 3.0, 4.0, 4.5, 5.0 and 5.5 W) on 16 calves showed that response latencies decreased as power increased up to 4.5 W, after which no further change occurred. In Experiment 3, the repeatability of the method was evaluated on nine measures with the high power setting (4.5 W). The coefficient of variation associated with repetition of the measures was 36%. In general, we found little change in response latencies with repeated use of the laser, except that responses on the second test tended to be shorter. Experiment 4 showed that ambient temperatures between 16 degrees C and 27 degrees C did not affect response latencies, but these were longer at temperatures of 7 degrees C. We suggest that the method is a useful way of measuring cattle's sensitivity to nociception as the animals need not be restrained and the distance to the animal need not be closely controlled. However, to obtain accurate, valid and reliable measures it is necessary to use a high power setting (4.5 W) and take at least three consecutive measures of the response latency.     

Enzyme histochemistry of rat mast cell tryptase

Fixation and staining conditions for rat mast cell tryptase and its histochemical distribution in different rat tissues were investigated. Prostate, skin, lung, gut, stomach and salivary glands were fixed in either aldehyde or Carnoy fixatives and then frozen or embedded in paraffin wax. Preservation of tryptase enzymic activity against peptide substrates required aldehyde fixation and frozen sectioning. Of the peptide substrates examined, z-Ala-Ala-Lys-4-methoxy-2-naphthylamide and z-Gly-Pro-Arg-4-methoxy-2-naphthylamide proved the most effective for the demonstration of tryptase. Double staining by enzyme cytochemistry followed by immunological detection of tryptase showed that, in all tryptase-containing mast cells, the enzyme is at least in part active. Conventional dye-binding histochemistry was used to confirm the identity of mast cells. Aldehyde-fixed mucosal mast cells required a much shorter staining time with Toluidine Blue if tissue sections were washed directly in t-butyl alcohol. Double staining by enzyme cytochemistry and dye binding showed that tryptase is absent from mucosal and subepidermal mast cells, which are also smaller in size and appear to contain fewer granules than connective tissue mast cells. This study demonstrates that rat mast cell tryptase, unlike tryptases in other species, is a soluble enzyme. It is stored in an active form and is absent from some mast cell subpopulations in mucosa, skin and lung. © 1998 Chapman & Hall     

Characterization of tissue tolerance to iron by molecular markers in different lines of rice

Ferrous iron (Fe2+) toxicity is a major disorder in rice prod uction on acid, flooded soils. Rice ( Oryza sativa L.) genotypes differ widely in tolerance to Fe2+ toxicity, which makes it possible to bre ed more tolerant rice varieties. Tissue tolerance to higher iron concentrations in plants has been considered to be important to Fe2+ tolerance in ri ce. Segregation for leaf bronzing and growth reduction due to Fe2+ to xicity was observed in a doubled haploid (DH) population with 135 lines derived from a Fe2+ tolerant japonica variety, Azucena, and a sensitive indic a variety, IR64 in a solution culture with Fe2+ stress condition at a Fe2+concentration of 250 mg L-1 at pH 4.5. To better understand the mechanism of tissue tolerance, Leaf Bronzing Index (LBI), total iron concentration in shoot tissue and the enzymes of ascorbate peroxidase (AP), dehydroascorbate reductase (DR) and glutathione reductase (GR), and concentrations of ascorbate (AS) and dehydroascorbate (DHA), which are involved in the ascorbate-specific H2O2-scavenging system, were determined for the population under Fe2+ stress. A non-normal distribution of LBI was found. About 38 lines showed no bronzing, while the lines with non-zero LBI values ranged from 0.05 to 0.85 and showed a normal distribution. The other parameters measured showed normal distribution. The total iron concentrations in the 38 tolerant lines ranged from 1.76 mg Fe g-1 to 4.12 mg Fe g-1 and was in a similar range as in the non-tolerant genotype (2.04 – 4.55 mg Fe g-1). No significant differences in the activities of the enzymes were found between the parents under normal culture, but remarkably higher Fe2+ induced enzyme activities were observed in the tolerant parent. AS was similar between the parents under both normal and Fe2+ stress, but its concentration was sharply decreased under Fe2+ stress. DHA was much lower in the tolerant parent than in the sensitive parent under Fe2+ stress. Single locus analysis and interval mapping analysis based on 175 molecular markers revealed that the interval flanked by RG345 and RZ19 on chromosome one was an important location of gene(s) for Fe2+ tolerance. The ascorbate-specific system for scavenging Fe2+-mediated oxygen free radicals may be an important mechanism for tissue Fe2+ tolerance. A gene locus with relative small effect on root ability to exclude Fe2+ was also detected.