Activation of lignin and manganese peroxidase excretion in Phanerochaete chrysosporium by fungal extracts

Summary Lignin (LiP) and manganese peroxidase (MnP) excretion by Phanerochaete chrysosporium INA-12 was significantly increased in response to fungal extract supplementation. LiP and MnP production was increased 1.7- and 1.8-fold, respectively, with fungal extracts from agitated pellet cultures of strain INA-12, namely fungal extracts P₆ and P₄. In cultures supplemented with a fungal extract harvested from static cultures of strain INA-12 (fungal extract S₄), LiP and MnP production was increased 1.8- and 1.6-fold, respectively. Succinate dehydrogenase activity, a mitochondrial marker, was significantly enhanced (2.7-fold) in cultures with the addition of fungal extracts. Correspondence to: M. Asther     

Characterization of the multiple forms of β-xylanase produced by a Streptomyces sp. growing on lignocellulose

Summary Streptomyces sp. strain EC10 degraded efficiently the hemicellulose fraction of wheat straw. Three forms of -xylanases detected in the culture filtrate were purified by precipipation with ammonium sulphate, chromatography on DEAE-Sephadex A-50 and gel filtration on Sephadex G-100. The three purified enzymes (X _ia , X _ib and X _ii ) were homogeneous by polyacrylamide gel electrophoresis. The enzymes were typical non-debranching endo--xylanases (1,4--d-xyla xylanohydrolases; E.C.3.2.1.8) with respective relative molecular weights of 32,000, 22,000 and 21,000 and isoelectric points of 6.8, 8.9 and 5.2. The enzymes were highly specific for xylans and showed optimal activity at pH 7.0–8.0 and 60°C. The preparations were completely free from cellulolytic activity (endoglucanase) and showed high thermal stability. No synergy between the three enzymes was detected for complete xylan hydrolysis of deacetylated arabino- and glucuronoxylans.Offprint requests:to: M. J. Penninckx     

Production of Cell Wall-Degrading Enzymes by the Phytopathogenic Fungus Sclerotinia sclerotiorum

The range of polysaccharide-degrading enzymes and glycosidases formed by the phytopathogenic fungus Sclerotinia sclerotiorum was monitored following growth on 16 carbohydrate substrates. Endo- and exoenzymes capable of degrading cellulosic, hemicellulosic, and pectinolytic polysaccharides were secreted. Pectinolytic activities were produced constitutively on all of the substrates tested. Cellulolytic enzymes were not induced in simple sugar (i.e., glucose or xylose) media. Polysaccharide growth substrates and cellulase inducers increased all of the enzyme activities tested. Gel filtration analysis revealed the appearance of new molecular forms of pectinase, β-xylosidase, and cellobiosidase during induction on pectin and carboxymethyl cellulose media.     

In situ spectroradiometric estimation of microalgal biomass in first-year sea ice

Summary First- and multi-year sea ice are colonized by microalgae, whose biomass modifies the spectral distribution of underice downwelling irradiance. It is proposed that an index of algal biomass in the first-year ice may be derived from the ratio of underice irradiance at a wavelength where absorption by chlorophyll a is high to a wavelength where absorption by the photosynthetic pigments is low and transmission through the ice is high. In southeastern Hudson Bay (Canadian Arctic), the irradiance ratio (671540 nm) accounts for 55% of the variance in chlorophyll a, indicating that the in situ biomass of algae in first-year ice can be estimated from spectral measurements of underice downwelling irradiance.Contribution to the programme of GIROQ (Groupe interuniversitaire de recherches océanographiques du Québec)     

Responses of antioxidants and lipid peroxidation in mussels to oxidative damage exposure.

1. The aim of this work was to evaluate the relationships between free radical scavengers and lipid peroxidation in the common mussel Mytilus edulis. 2. Mussels were exposed to compounds known for their ability to produce free radicals (carbon tetrachloride, CCl4) and reactive oxygen species via redox cycling (menadione) and the effects on digestive gland, gills and remaining tissues were studied. 3. Lipid peroxidation parameters and the status of free radical scavengers (glutathione, vitamins A, E and C) were affected more by exposure to menadione than to CCl4. 4. The observed changes in the free radical scavengers content are indicative of a role in detoxication of damaging reactive species.     

Oxysterol activation of arachidonic acid release and protaglindin E2 biosynthesis in NRK 49F cells is partially dependent on protein kinase activity

We previously demonstrated that the oxysterol potentiation of arachidonic acid release and prostaglandin biosynthesis induced by foetal calf serum activation of normal rat kidney (NRK) cells (fibroblastic clone 49F) was not related to a direct effect of oxysterols on cell free Ca²⁺ level. Since both Ca²⁺ variations and protein C are involved in arachidonic acid release in some models, we looked for a possible modulation by protein C in the oxysterol effect on arachidonic acid release. We show that when the phorbol ester 12-O-tetradecanoyl-phorbol-13acetate (TPA), a protein kinase C activator, was added to the culture medium, the oxyterol effect on arachidonic acid release and prostaglandin synthesis clearly increased. Moreover, the effect of TPA was dose-dependent and TPA EC₅₀ (4 × 10⁻⁹ M) was unchanged in the presence of the oxysterol. Preincubation of cells with TPA for 24 h prevented the arachidonic acid release induced by TPA alone, whereas the oxysterol effect was decreased but not abolished. In the absence of serum, TPA and ionomycin added together induced the same noticeable (arachidonic acid) release and PGE₂ synthesis as serum alone. Nevertheless, the potentiating effect of cholest-5-ene-3β,25-diol was much higher when serum itself was used to activate NRK cells than it was in the present serum-mimicking experimental conditions. Thus, the presence of growth factors is probably required to obtain a full oxysterol effect. We conclude that the oxysterol effect was synergistic with, but not fully dependent on, protein kinase C and Ca²⁺ ion fluxes, therefore oxysterols could affed earlier events triggered by serum growth factor binding to their cell membrane receptors.     

Influence of inoculum age on hybridoma culture kinetics

To determine the influence of the inoculum age on the kinetics of hybridoma growth and metabolism, spinner flasks have been inoculated with cells previously propagated in T flasks for 43, 52, 62 and 71 hr respectively. Increasing the age of the inoculum is found to result in a longer lag phase, in a lower maximum specific growth rate and in a reduced maximal cell density. During the growth phase specific rates of glucose and glutamine uptake and of ammonia and lactate production are similar. However, with the older inoculum, much higher metabolic activities are observed during the lag phase. The production of antibodies is delayed with increasing inoculum age, but the final antibody concentrations are similar, which indicates a higher specific antibody production rate when inoculating with older cells.     

Purine nucleosides and nucleotides stimulate proliferation of a wide range of cell types

Summary Presumptive astrocytes isolated from 10-day white Leghorn chick embryos, Factor VIII-positive human brain capillary endothelial cells, meningeal fibroblasts from 10-day chick embryos, Swiss mouse 3T3 cells, and human astrocytoma cell lines, SKMG-1 and U373, were rendered quiescent when placed in culture medium that contained 0 or 0.2% serum for 48 h; their proliferation was markedly reduced and they incorporated [³H]thymidine at a low rate. [³H]Thymidine incorporation and cell proliferation were induced in all types of cells by addition of guanosine, GMP, GDP, GTP, and to a lesser extent, adenosine, AMP, ADP or ATP to the culture medium. The stimulation of proliferation by adenosine and guanosine was abolished by 1,3-dipropyl-7-methylxanthine (DPMX), an adenosine A₂ receptor antagonist, but not by 1,3,-dipropyl-8-(2-amino-4-chorophenyl)xanthine (PACPX), an A₁ antagonist. Stimulation of proliferation by the nucleotides was not abolished by either DPMX or PACPX. The P₂ receptor agonists,α,β-methyleneATP and 2-methylthioATP, also stimulated [³H]thymidine incorporation into the cells with peak activity at approximately 3.5 and 0.03 nM, respectively. These data imply that adenosine and guanosine stimulate proliferation of these cell types through activation of an adenosine A₂ receptor, and the stimulation of cell proliferation by the nucleotides may be due to the activation of purinergic P_2y receptors. As the primary cultures grew older their growth rate slowed. The capacity of the purine nucleosides and nucleotides to stimulate their growth diminished concomitantly. The 3T3 cells showed neither decreased growth with increased passages nor reduced response to the purines. In contrast, although the doubling time of the immortalized human astrocytoma cell lines SKMG-1 and U373 remained constant, the responsiveness to purinergic stimulation of the U373 cells decreased but that of the SKMG-1 did not. These data are compatible with a decrease in the number, or the ligand-binding affinity of the purinergic receptors, or a decreased coupling of purinergic receptors to intracellular mediators in primary cells aged in tissue culture.     

Analysis of Gene Evolution: the software AGE

The software AGE (Analysis of Gene Evolution) has been developedboth to study a genetic reality, i. e. the identification ofstatistical properties in genes (e.g. periodicities), and tosimulate this observed genetic reality, by models of molecularevolution. AGE has two types of models: (i) models of sequencecreation from oligonucleotides: concatenation model in seriesof an oligonucleotide, independent (or Markov) mixing modelof oligonucleotides according to given probabilities (or a Markovmatrix); (ii) models of sequence evolution from created sequences:insertion/deletion process of (mono, di, tri)nucleot-ides, basemutation process. The study of a reality and the developmentof simulation models are based on several new algorithms: approximatedsimulation and exact calculus to compute various autocorrelationfunctions, Fourier transformation of autocorrelation curves,recognition of a curve form, etc. AGE is implemented on IBMor compatible microcomputers and can be used by biologists withoutany computer knowledge to identify statistical properties intheir newly determined DNA sequence and to explain them by modelsof molecular evolution.