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PubMed2006

Phylogenetic diversity of acidophilic sporoactinobacteria isolated from various soils.

Spore forming actinobacteria (sporoactinobacteria) isolated from soils with an acidic pH in Pinus thunbergii forests and coal mine waste were subjected to taxonomic characterization. For the isolation of acidophilic actinobacteria, acidified starch casein agar (pH adjusted to 4-5) was used. The numbers of actinobacteria growing in acidic media were between 3.2 x 10(4) and 8.0 x 10(6) CFU/g soil. Forty three acidophilic actinobacterial strains were isolated and their 16S rDNA sequences were determined. The isolates were divided into eight distinctive phylogenetic clusters within the variation encompassed by the family Streptomycetaceae. Four clusters among them were assigned to the genus Streptacidiphilus, whereas the remaining four were assigned to Streptomyces. The clusters belonging to either Streptomyces or Streptacidiphilus did not form monophyletic clade. The growth pH profiles indicated that the representative isolates grew best between pH 5 and 6. It is evident from this study that acidity has played a critical role in the differentiation of the family Streptomycetaceae, and also that different mechanisms might have resulted in the evolution of two groups, Streptacidiphilus (strict acidophiles) and neutrotolerant acidophilic Streptomyces. The effect of geographic separation was clearly seen among the Streptacidiphilus isolates, which may be a key factor in speciation of the genus.

Coal↗

PubMed1994

Effect of feeding ponderosa pine needle extracts and their residues to pregnant cattle.

Solvents including ethanol, methylene chloride, acetone, hexane, and hot and cold water were used in separate procedures to extract needles and bark from ponderosa pine (Pinus ponderosa L.) to determine which system was optimal for the extraction of abortifacient compound(s). The extracts and residues therefrom were fed by gavage to pregnant cows beginning at day 250 of gestation. After methylene chloride extraction, the pine needle residue fed by gavage failed to abort 4 cows. Subsequently, 4 cows fed the methylene chloride extractable material, adsorbed on ground hay, but free of methylene chloride, aborted (days 253-260 of gestation) and all 4 cows retained fetal membranes. Only 2 of 6 cows fed the pine needle residue after ethanol extraction aborted (days 260 and 261 of gestation) and both retained their fetal membranes, while the other 4 cows calved normally. Hot and cold water extracts did not cause abortion, whereas the extracted plant residue did. One cow fed the plant residue after acetone extraction calved normally as did 1 of 2 cows fed residue after hexane extraction. One cow fed hexane extract absorbed on ground hay also calved normally. These results demonstrate that abortifacient compound(s) present in pine needles are most efficiently extracted by methylene chloride.

Abortifacient Agents↗

PubMed2000

Evaluation of enzymes produced from yeast.

Pichia pinus was found to be capable of growing on mango wastes, producing pectinase (pectin lyase, EC-4.2.2.10) and lactase (beta-galactosidase, EC-3.2.1.23) enzymes. The two enzymes were successively purified by precipitation with ammonium sulfate followed by chromatography on Sephadex G-120. The purification procedure provided 1,846 and 929 fold purification with 20.6 and 24% yield recovery of pectinase and lactase, respectively. the km value of pectinase was 0.33% for pectin at pH 4.5 and that for lactase was 0.166% for lactose at pH 7.0. The purified enzymes, pectinase and lactase are stable up to 50 degrees C for 60 and 45 min, respectively, with 20 and 35% loss of their activity. Gel filtration on Sephadex G-200 indicated that the molecular weights of the purified pectinase was 90 x 10(3) Dalton and of lactase 115 x 10(3) Dalton. On the basis of the evaluation tests done, the enzymes were considered to have a potential technological interest as treating mango pastes (residues left after mango juice preparation) with the two prepared enzymes resulted in an increase of the colour intensity, total carbohydrate content and juice yield. Treating milk with the purified lactase also showed an increase in the total carbohydrate and reducing sugar produced.

Chromatography, Gel↗

PubMedDate not supplied

Separation and characterization of needle and xylem maritime pine proteins.

Two-dimensional gel electrophoresis (2-DE) and image analysis are currently used for proteome analysis in maritime pine (Pinus pinaster Ait.). This study presents a database of expressed proteins extracted from needles and xylem, two important tissues for growth and wood formation. Electrophoresis was carried out by isoelectric focusing (IEF) in the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second. Silver staining made it possible to detect an average of 900 and 600 spots on 2-DE gels from needles and xylem, respectively. A total of 28 xylem and 35 needle proteins were characterized by internal peptide microsequencing. Out of these 63 proteins, 57 (90%) could be identified based on amino acid similarity with known proteins, of which 24 (42%) have already been described in conifers. Overall comparison of both tissues indicated that 29% and 36% of the spots were specific to xylem and needles, respectively, while the other spots were of identical molecular weight and isoelectric point. The homology of spot location in 2-DE patterns was further validated by sequence analysis of proteins present in both tissues. A proteomic database of maritime pine is accessible on the internet (http://www.pierroton.inra.fr/genetics/2D/).

Acrylic Resins↗

PubMed2002

Fourier Transform-Raman spectroscopic study of natural resins of archaeological interest.

Resins from several different genera are studied using Fourier transform (FT)-Raman spectroscopy. Tree resins can be broadly divided into those that contain diterpenoid components and those that contain triterpenoid components. The diterpenoid resins analyzed are from the genera Pinus, Cedrus, and Agathis (kauri resin) and the triterpenoid resins examined are samples from Pistacia, Boswellia (frankincense), and Commiphora (myrrh) genera. A protocol is developed to nondestructively distinguish diterpenoid and triterpenoid resins and to differentiate the genera within the two types. The effects of oxidation on the discrimination of the FT-Raman spectra are considered.

Archaeology↗

PubMed2004

Airborne fungal spores, pollen grains, and vegetable cells in routine Papanicolaou smears.

The objective was to demonstrate the presence of airborne fungal spores, pollen grains, and vegetable cells in smears and establish their significance. Microscopic examination was of smears stained by the Papanicolaou technique. We found several types of airborne spores (Alternaria, Exserohilum, Aspergillus, Cladosporium, Epicoccum, Curvularia, and Ascosmycete), pollen grains (Lilium, bullrush, Pinus, Acacia, hazelnut, and oak), and several types of vegetable cells. The existence of these structures may be due to intrinsic or to extrinsic contamination of the sample. Anemophilous fungi spores and pollen grains have been shown to possess great allergenic capacity. Pollen grains and vegetable cells may be mistaken for certain microorganisms or malignant cells.

Cytodiagnosis↗

PubMed1989

A two-dimensional electrophoresis procedure for single meristems of different forest species.

High resolution two-dimensional electrophoresis, with isoelectric focusing in the first dimension and electrophoresis in sodium dodecyl sulfate in thin acrylamide gels in the second dimension, has been applied to separate the proteins of single meristems (200-300 microns) from Sequoiadendron giganteum, Sequoia sempervirens, Pseudotsuga menziesii, Picea abies, Pinus pinaster, Eucalyptus gunnii and Populus nigra. The technique may prove an efficient tool for studying inter- and intraclonal differences in meristem cultures of forest species, especially when referring to cloning from meristems of selected trees in relation to the rejuvenation process.

Electrophoresis, Gel, Two-Dimensional↗

PubMedDate not supplied

Electrophoretic analysis of seed storage proteins from gymnosperms.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), pore gradient gel electrophoresis (PGGE) followed by SDS-PAGE and Western blot analysis were used to characterize the seed storage proteins from seven gymnosperm species from the families Pinaceae (Abies alba, Cedrus atlantica and Picea abies), Cupressaceae (Biota orientalis, Chamaecyparis lawsoniana and Cupressus arizonica), and Taxaceae (Taxus baccata). SDS-PAGE and PGGE X SDS-PAGE indicate the presence of proteins with characteristics similar to the 7S globulins in all the species studied. Antibodies to a 7S globulin subunit from Pinus pinaster cross-reacted with homologous polypeptides from Pinaceae species, but not with corresponding subunits from species belonging to other families. Also detected in each of the studied species, with the exception of A. alba and T. baccata, were those of C. atlantica may be extracted by saline buffer, while the remainder require a dissociating agent. Antibodies raised against the small subunit from P. pinaster 11S protein recognized only the corresponding polypeptides from Pinaceae species. Overall, these results help clarify our knowledge of gymnosperm seed storage proteins.

Animals↗

PubMed1988

Genetic control of methanol utilization in yeasts.

Considered are our own data and those found in literature on the properties of yeast mutants impaired in their ability to utilize methanol as sole carbon and energy source; hypotheses about the role of alcohol oxidase and citrate synthase in biogenesis of peroxisomes are proposed. It has been proved that formaldehyde reductase participates in the control of the formaldehyde level in the cell. Properties of mutants defective in the catabolite repression and inactivation of enzymes of methanol metabolism are described. The existence of several autonomous mechanisms of the catabolite repression of alcohol oxidase has been shown. It has been found, that the induction of glyoxysomal enzymes of C2-metabolism is repressed by methanol in the ecr1 mutant of Pichia pinus with the affected repression of alcohol oxidase by ethanol. Data are presented on the regulatory properties of the recently discovered acidification system of the medium induced by methanol. Such acidification occurs due to symport extrusion of protons and formate anions from the cells.

Biotechnology↗

PubMed1991

Mycoflora associated with five species of freshwater leeches.

Twenty-two species belonging to 11 genera of zoosporic fungi were collected from 10 freshwater sites and from five species of leeches on sesame and hemp seeds, maize grains and Pinus pollen grains as baits at 22 degrees C. Dictyuchus, Achlya, Pythium, Allomyces and Saprolegnia were the most common genera. The highest population of zoosporic fungi was estimated from Batracobdelloides tricarinata, the lowest from Barbronia assiuti. In addition fifty-three species and 1 variety representing 23 terrestrial fungal genera were identified on glucose (17 genera and 41 species) and cellulose (20 genera and 34 species + 1 variety) Czapek's Dox agar at 28 degrees C. The most common species on the two media were Aspergillus niger, A. fumigatus, A. flavus, Trichoderma harzianum, Gibberella fujikuroi and Penicillium chrysogenum. Some fungi were common only on cellulose agar medium such as Botryotrichum atrogriseum, Chaetomium bostrychodes, Dactylella alaskana and Drechslera halodes. Batracobdelloides tricarinata was the richest with terrestrial fungi whereas Salifa delicata was the poorest. Five leech species, namely Alboglossiphonia polypompholyx, Batracobdelloides tricarinata, Helobdella conifera, Barbronia assiuti and Salifa delicata were used during this investigation.

Animals↗

PubMed2004

Pycnogenol and vitamin E inhibit ethanol-induced apoptosis in rat cerebellar granule cells.

Pycnogenol (PYC), a patented combination of bioflavonoids extracted from the bark of French maritime pine (Pinus maritima), scavenges free radicals and promotes cellular health. The protective capacity of PYC against ethanol toxicity of neurons has not previously been explored. The present study demonstrates that in postnatal day 9 (P9) rat cerebellar granule cells the antioxidants vitamin E (VE) and PYC (1) dose dependently block cell death following 400, 800, and 1600 mg/dL ethanol exposure (2) inhibit the ethanol-induced activation of caspase-3 in the same model system; and (3) reduce neuronal membrane disruption as assayed by phosphatidylserine translocation to the cell surface. These results suggest that both PYC and VE have the potential to act as therapeutic agents, antagonizing the induction of neuronal cell death by ethanol exposure.

Animals↗

PubMed2004

Protective mechanisms of pycnogenol in ethanol-insulted cerebellar granule cells.

Pycnogenol (PYC), a patented combination of bioflavonoids extracted from the bark of French maritime pine (Pinus maritima), inhibits apoptosis and necrosis of developing neurons exposed acutely to ethanol (EtOH). The present study shows that the protective mechanisms of PYC in EtOH-exposed postnatal day 9 cerebellar granule cells (P9 CGCs) include (1) reduction of reactive oxygen species (ROS) production; (2) counteraction of suppressed copper/zinc superoxide dismutase (Cu/Zn SOD) and glutathione peroxidase/reductase (GSH-Px/GSSG-R) system activities; (3) upregulation of Cu/Zn SOD protein expression; (4) mitigation of the EtOH-mediated exacerbation of catalase (CAT) activity; and, (5) specific binding and inhibition of active caspase-3. These results indicate that the mechanisms by which PYC antagonizes EtOH-induced oxidative stress include oxidant scavenging and modulation of endogenous, cellular proteins. Using findings from the present and previous studies, a model delineating the mechanisms of EtOH effects on the system of antioxidant enzymes in developing CGCs is presented.

Animals↗

PubMed2005

Repellency and toxicity of aromatic plant extracts against the mosquito Culex pipiens molestus (Diptera: Culicidae).

The insecticidal activities of essential oil extracts from leaves, flowers and roots of aromatic plants against fourth-instar larvae of the mosquito Culex pipiens molestus Forskal were determined. Extracts of Foeniculum vulgare Mill were the most toxic, followed by those of Ferula hermonis Boiss, Citrus sinensis Osbeck, Pinus pinea L, Laurus nobilis L and Eucalyptus spp with LC50 values of 24.5, 44.0, 60.0, 75.0, 117.0 and 120.0 mg litre(-1), respectively. Combination tests between the LC50 and the maximum sub-lethal concentration (MSLC) were determined. Over 20 major components were identified in extracts from each plant species tested. Five essential oils and nine pure components were studied for their repellency against mosquito bites. Terpineol and 1,8-cineole were the most effective against Culex pipiens molestus bites offering complete protection for 1.6 and 2 h, respectively.

Animals↗

PubMed2005

Enhancement of attraction and trap catches of the old-house borer, Hylotrupes bajulus (Coleoptera: Cerambycidae), by combination of male sex pheromone and monoterpenes.

Monoterpenoid compounds extracted from wood of Scots pine, Pinus sylvestris L, synergized the attraction of the old-house borer, Hylotrupes bajulus (L), to the male pheromone (3R)-3-hydroxy-2-hexanone ((3R)-ketol) + 1-butanol. Glasshouse experiments using ground traps baited with extracts derived from Scots pine wood or the monoterpenes (+)-alpha-pinene, (-)-verbenone, (-)-trans-pinocarveol and (+)-terpinen-4-ol attracted significantly more H bajulus females, but caught fewer of them, than the synthetic pheromone mixture alone. Traps baited with higher concentrations of the monoterpene mixture attracted fewer females than those baited with lower dosages, whilst very high concentrations of the mixture (6-10 vials) caught no insects. However, a combination of (3R)-ketol + 1-butanol or (+/-)-3-ketol + 1-butanol with monoterpenes resulted in the capture of significantly more females than either the sex pheromone or the monoterpene mixture alone. Traps baited with a blend of the male's sex pheromone or the monoterpenes attracted significantly more, but caught fewer, males than females. Here again, a combination of the above blends enhanced the attraction of males significantly when compared with the attractancy of either of the compounds/mixtures used alone.

Animals↗

PubMed2001

Development of a water-soluble preparation of emamectin benzoate and its preventative effect against the wilting of pot-grown pine trees inoculated with the pine wood nematode, Bursaphelenchus xylophilus.

Water-soluble preparations have been investigated to develop a trunk injection agent based on the poorly water-soluble anti-nematode emamectin benzoate. Following tests on the phytotoxicity of some solvents and solubilizers and demonstration of the ability of some solubilizers to dissolve emamectin benzoate in water, acetone + methanol was selected as the solvent and Polysorbate 80 as the solubilizer. This water-soluble preparation of emamectin benzoate prevented the wilting of pot-grown 4-year-old trees of the Japanese black pine, Pinus thunbergii, artificially inoculated with the pine wood nematode, Bursaphelenchus xylophilus, at a dose of 20 g emamectin benzoate per cubic metre of pine tree.

Animals↗

PubMed2001

The anxiolytic effect of Sho-ju-sen, a Japanese herbal medicine, assessed by an elevated plus-maze test in mice.

Sho-ju-sen (SK), a Japanese herbal medicine with a nourishing tonic action, is composed of a water extract of Kumazasa leaves (Sasa kurinensis Makino et Sibata) (SS), and ethanol extracts of Japanese red pine needles (Pinus densiflora Sieb. et Zucc) (PN) and Ginseng roots (Panax ginseng C. A. Meyer) (PX) in the ratio 8:1:1. In this study, an elevated plus-maze test in mice was carried out to assess whether SK had an anxiolytic effect. No significant change was observed in either the plus-maze or activity test following a single administration of SK (10 and 20 mL/kg p.o.). However, mice allowed a free intake of SK (10% solution) for 5 days and longer showed a significant prolongation of the time spent in the open arms (an anxiolytic effect), as long as that caused by the benzodiazepine anxiolytic diazepam (1 mg/kg p.o.). SK (1%, 3% and 30% solutions for 7 days) tended to develop the anxiolytic effect. Of the constituents of SK, SS (8% solution), but not PN (1% solution) or PX (1% solution), resulted in the anxiolytic effect. Except for a slight acceleration in the motor activity by PN (1% solution), no significant change in the motor activity was produced by any treatment with SK, SS or PX. The combined treatment of SK (10% solution) or SS (8% solution) with 1 mg/kg diazepam enhanced the anxiolytic effect. Flumazenil (0.1 mg/kg s.c.), a benzodiazepine receptor antagonist, alone did not change the time spent in the open arms. However, it completely reversed the anxiolytic effect of SK, SS and diazepam. The present results suggest that: (1) long-term treatment with SK develops an anxiolytic effect, (2) SS is the main constituent for the anxiolytic effect of SK, and (3) benzodiazepine receptors are involved in the anxiolytic effect of SK and SS.

Animals↗

PubMed2001

Antimicrobial activity of some Pacific Northwest woods against anaerobic bacteria and yeast.

Extracts of woods commonly used for animal bedding were tested for antimicrobial activity. Essential oils from Alaska cedar (Chamaecyparis nootkatensis), western juniper (Juniperus occidentalis) and old growth Douglas fir (Pseudotsuga menziesii) as well as methanol extracts of wood from these trees plus western red cedar (Thuja plicata) and ponderosa pine (Pinus ponderosa) were tested for antimicrobial activity against anaerobic bacteria and yeast. The test microbes included Fusobacterium necrophorum, Clostridium perfringens, Actinomyces bovis and Candida albicans which are common to foot diseases and other infections in animals. The essential oils and methanol extracts were tested using a standardized broth assay. Only extracts of Alaska cedar and western juniper showed significant antimicrobial activity against each of the microbes tested. The essential oil of Douglas fir did show antimicrobial activity against A. bovis at the concentrations tested. The methanol extracts of the heartwood of Douglas fir and the sapwood of ponderosa pine showed no antimicrobial activity. The major chemical components of western juniper (cedrol and alpha- and beta-cedrene) and Alaska cedar (nootkatin) were also tested. In western juniper, alpha- and beta-cedrene were found to be active components. Nootkatin showed activity only against C. albicans. The inhibitory activity in Alaska cedar oil was high enough to justify further efforts to define the other chemical components responsible for the antimicrobial activity.

Anti-Bacterial Agents↗

PubMed1993

Biosynthesis of monoterpenes: regio- and stereochemistry of (+)-3-carene biosynthesis.

Incubation of [1-3H1]geraniol with stem disks of Douglas fir (Pseudotsuga menziesii) and incubation of [1-3H1]geranyl pyrophosphate with both a soluble enzyme extract from Douglas fir and a partially purified preparation of (+)-3-carene synthase from lodgepole pine (Pinus contorta) resulted in the production of (+)-3-[3H] carene. Subsequent conversion of the product to car-3-en-5-one and to 4-isocaranone followed by base-catalyzed exchange of the alpha-hydrogens established that the 3H located at C1 in the geranyl substrate resided at C5 of (+)-3-carene. Incubation of the (+)-3-carene synthase preparation with (S)-[5-3H1, 4-14C]geranyl pyrophosphate resulted in the production of (+)-3-carene without loss of tritium, indicating that the 5-proR hydrogen is eliminated during cyclopropyl ring closure. Analysis of the conformational requirements for this 1,3 elimination involving the 5-proR hydrogen suggested that cyclopropyl ring formation occurs via a (4S)-alpha-terpinyl cation derived from the anti-endo cyclization of a (3S)-linalyl pyrophosphate intermediate. Kinetic analyses of the conversion of (1Z,3R)-[1-3H1]linalyl pyrophosphate, (1Z, 3S)-[1-3H1]linalyl pyrophosphate and [1-3H1]geranyl pyrophosphate by (+)-3-carene synthase revealed that the velocity of the reaction with the (3S)-linalyl enantiomer was 25-fold greater than the velocity with the (3R)-enantiomer and twice that of the natural substrate, geranyl pyrophosphate, thereby confirming this stereochemical prediction and also indicating that the cyclization of the linalyl intermediate is faster than the coupled isomerization and cyclization of the geranyl substrate. From these results, a model that details the regio- and stereochemistry of the enzymatic conversion of geranyl pyrophosphate to (+)-3-carene is proposed.

Bicyclic Monoterpenes↗