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PubMed2003

Pilot study on the clinical effects of dietary supplementation with Enzogenol, a flavonoid extract of pine bark and vitamin C.

Flavonoids are naturally occurring plant compounds with established in vitro antioxidant properties and potential cardioprotective effects. We carried out a 12-week pilot study on the effects of dietary supplementation with an extract of bioflavonoids prepared from the bark of Pinus radiata trees [Enzogenol] containing added vitamin C. Data was collected from 24 healthy subjects aged between 55-75 years at baseline and at 6 and 12 weeks and included, routine biochemical and haematological indices, and anthropometric, blood pressure, forearm blood flow and haemorheological measurements. Enzogenol supplementation at a dosage of 480 mg/day of pine bark extract and 240 mg/day vitamin C did not result in changes in any biochemical or haematological indice and was associated with a significant reduction in the means of body weight, percentage body fat, systolic blood pressure and plasma viscosity. Basal and hyperaemic blood fl ow in forearm resistance vessels measured by plethysmography increased significantly during the study. The findings of this pilot study indicate that dietary supplementation with Enzogenol is safe and well tolerated and is associated with a number of beneficial effects on a range of established cardiovascular risk factors. These changes need to be validated by a placebo-controlled study but are consistent with other studies that have reported beneficial clinical effects following supplementation with bioflavonoids.

Aged↗
PubMed2003

Pycnogenol inhibits the release of histamine from mast cells.

Oxygen derived free radicals are now increasingly regarded as a primary force of tissue destruction and also have the ability to release histamine from mast cells. Pycnogenol is an extract of the bark of French maritime pine (Pinus pinaster) containing bioflavonoids with a potent ability to scavenge free radicals. Therefore Pycnogenol was investigated for inhibition of histamine release from rat peritoneal mast cells. In addition, its effects were compared with sodium cromoglycate, a known inhibitor of histamine release from the mast cell. Rat peritoneal mast cells were isolated and purified by differential centrifugation and cells pooled from 3-4 animals were suspended at approximately 10(6) cells/mL buffered salt solution. Histamine release was induced by compound 48/80 or the calcium ionophore A-23187 and estimated from supernatant following extraction and by fluorimetric methods. Pycnogenol produced a concentration dependent inhibition of histamine release induced by the two secretagogues. Its inhibitory effect on mast cell histamine release was favourably comparable to sodium cromoglycate.

Animals↗
PubMed2003

Pycnogenol prevents haemolytic injury in G6PD deficient human erythrocytes.

Glucose6 phosphate dehydrogenase (G6PD) deficiency is the most common X-linked disorder of human erythrocytes where cells have inadequate capacity to destroy peroxides and high susceptibility towards haemolytic changes. Pycnogenol is a proprietary dry extract of the French Maritime pine (Pinus pinaster) bark with high ability to scavenge free radicals. In the present study we have investigated if Pycnogenol can protect G6PD deficient erythrocytes against haemolytic cell damage. Venous blood samples were obtained from six subject of Mediterranean origin with known G6PD deficiency which was also confirmed with standard techniques. Erythrocyte haemolysis in the presence and absence of Pycnogenol was induced either with tert-butylhydroperoxide (t-BHP) or quinine and the haemoglobin release in the supernatant was determined by recording the optical density at 540 nm in a Shimadzu spectrophotometer. Our results have shown that Pycnogenol has protective action against a Xenobiotic chemical induced haemolysis in G6PD deficient human erythrocytes.

Erythrocytes↗
PubMed2004

The effect of Pycnogenol on fluoride induced rat kidney lysosomal damage in vitro.

Pycnogenol (PYC) is a procyanidin-rich extract of the bark of French maritime pine (Pinus pinaster) with a potent ability to scavenge free radicals. Lysosome-rich fractions from rat renal cortices were incubated with varying amounts of PYC and challenged with noxious doses of fl uoride. Controls were also included. The release of N-acetyl-beta-D-glucosaminidase (NAG) isozymes in the supernatant was estimated by spectrophotometric methods. The protein content of the renal cortex was also determined. Our results show that fluoride in unhealthy doses can cause a concentration dependent release of N-acetyl-beta-D-glucosaminidase (NAG) isozymes from the renal lysosomes. This may be related to its known ability to initiate free radical formation or direct damaging effects on the lysosomal membrane. As a blend of bio flavonoids pycnogenol has a potent ability to scavenge free radicals. In our study PYC was effective in preventing fluoride induced release of NAG isozymes from the renal lysosomes.

Animals↗
PubMed2004

An antidepressant effect of Sho-ju-sen, a Japanese herbal medicine, assessed by learned helplessness model in mice.

The antidepressant effect of Sho-ju-sen, a Japanese herbal medicine composed of extracts of three herbs; kumazasa leaf (Sasa Kurinensis Makino et Sibata), Japanese red pine leaf (Pinus densiflora Sieb. et Zucc) and ginseng radix (Panax ginseng C.A. Meyer), was assessed using a learned helplessness model in mice. The learned helplessness was produced by presenting 120 unavoidable/inescapable shocks for 3 days to the mouse in a shuttle box, and the avoidance training was carried out on day 4. Compared with the control group given tap water, free consumption of Sho-ju-sen (1%, 3% and 10%) for 21 days resulted in a significant amelioration of the response rate at 1% and 3%, and both the response rate and % avoidance at 10%. Although Sho-ju-sen (10%) caused no significant effect following the 7-day intake, it ameliorated the response rate following the 14-day intake. The extract of Japanese red pine leaf, but not kumazasa leaf or ginseng radix, mildly improved the response rate. Learned helplessness was significantly and dose-dependently reduced by imipramine (10 and 30 mg/kg i.p.), while only mildly by diazepam (1 mg/kg p.o.). These results suggest that a long-term consumption of Sho-ju-sen is effective for the amelioration of depression, and the effectiveness is derived mainly from the extract of Japanese red pine leaf.

Animals↗
PubMed2005

Antimicrobial activity of Pycnogenol.

Pycnogenol, a standardized extract of Pinus pinaster bark, was tested for its antimicrobial activity against 23 different pathogenic prokaryotic (gram-positive and gram-negative) and eukaryotic (yeast and fungi) microorganisms. Pycnogenol inhibited the growth of all the tested microorganisms in minimum concentrations ranging from 20 to 250 microg/mL. Thus, Pycnogenol in concentrations as low as 0.025% could counteract the growth of all the strains investigated in our study. These results conform with clinical oral health care studies describing the prevention of plaque formation and the clearance of candidiasis by Pycnogenol.

Anti-Bacterial Agents↗
PubMed2006

Antioxidant potential of six pine species.

The aim of the study was to evaluate the antioxidant efficacy of extracts obtained from six Pinus species (P. pinea, P. brutia, P. radiata, P. halepensis, P. attenuata, P. nigra) growing in natural forests in Southern Greece. Specimens of fresh, dry needles and pine bark were extracted and fractionated with a variety of organic solvents and the efficient concentration and their radical scavenging activity was evaluated by the Co(II)/EDTA induced luminol plateau chemiluminescence assay.

Antioxidants↗
PubMed1995

Xylem-specific gene expression in loblolly pine.

Two genes preferentially expressed in differentiating xylem of loblolly pine (Pinus taeda L.) were cloned from cDNA and genomic libraries and designated PtX3H6 and PtX14A9. Transcripts of PtX3H6 and PtX14A9 are very abundant in differentiating xylem, less abundant in needles, and very low or non-detectable in embryos and megagametophytes. PtX3H6 contains a putative signal peptide, a threonine-rich region, a proline-rich region, and a hydrophobic tail. Repeats of Pro-Pro-Pro-Val-X-X are similar to repeats found in proline-rich cell wall proteins. The amino acid compositions of PtX3H6 and PtX14A9 are similar to those of arabinogalactan proteins (AGPs). PtX14A9 contains an 8 amino acid sequence similar to amino terminal sequences of ryegrass, carrot and rose AGPs. Upstream sequences have been determined from genomic clones encoding PtX3H6 and PtX14A9. A 7 bp sequence found in the 5' flanking regions of both genes has previously been shown to be involved in the vascular-specific expression of GRP 1.8, a glycine-rich protein found in bean. The sequence is also present upstream of another glycine-rich protein from bean, GRP 1.0, and may be partially responsible for the xylem-specific expression of pTx3H6 and PtX14A9.

Amino Acid Sequence↗
PubMed1995

Genetic analysis of cinnamyl alcohol dehydrogenase in loblolly pine: single gene inheritance, molecular characterization and evolution.

The gene encoding the monolignol biosynthetic enzyme cinnamyl alcohol dehydrogenase (CAD, E.C. 1.1.1.195) can be expressed in response to different developmental and environmental cues. Control of Cad gene expression could involve either differential regulation of more than one Cad gene or, alternatively combinatorial regulation of a single Cad gene. In loblolly pine (Pinus taeda L.), we found several electrophoretic variants (allozymes) of CAD and a high level of heterozygosity (he = 0.46). Analysis of inheritance patterns of pine CAD allozymes gave segregation ratios that were consistent with Mendelian expectations for a single functional gene. The identity of the full-length Cad cDNA sequence was confirmed by alignment with peptide sequences obtained from purified active enzyme and by extensive similarity to Cad sequences from other species. Southern blot analysis of genomic DNA using the Cad cDNA as a hybridization probe gave simple patterns, consistent with our interpretation that pine Cad is a single-copy gene. Phylogenetic analysis and evolution rate estimates showed that Cad sequences are diverging less rapidly in the gymnosperms than in the angiosperms. The Cad mRNA was present in both lignifying tissues and a non lignifying tissue (the megagametophyte) of pine. The presence of a single gene suggests that different regulatory mechanisms for a single Cad gene, rather than differential regulation of several genes, can account for its expression in response to different cues.

Alcohol Oxidoreductases↗
PubMed2004

Plant resistance to cold stress: mechanisms and environmental signals triggering frost hardening and dehardening.

This introductory overview shows that cold, in particular frost, stresses a plant in manifold ways and that the plant's response, being injurious or adaptive, must be considered a syndrome rather than a single reaction. In the course of the year perennial plants of the temperate climate zones undergo frost hardening in autumn and dehardening in spring. Using Scots pine (Pinus sylvestris L.) as a model plant the environmental signals inducing frost hardening and dehardening, respectively, were investigated. Over 2 years the changes in frost resistance of Scots pine needles were recorded together with the annual courses of day-length and ambient temperature. Both act as environmental signals for frost hardening and dehardening. Climate chamber experiments showed that short day-length as a signal triggering frost hardening could be replaced by irradiation with far red light, while red light inhibited hardening. The involvement of phytochrome as a signal receptor could be corroborated by respective night-break experiments. More rapid frost hardening than by short day or far red treatment was achieved by applying a short period (6 h) of mild frost which did not exceed the plant's cold resistance. Both types of signals were independently effective but the rates of frost hardening were not additive. The maximal rate of hardening was - 0.93 degrees C per day and frost tolerance of less than < - 72 degrees C was achieved. For dehardening, temperature was an even more effective signal than day-length.

Freezing↗
PubMed2005

Enterobacter cloacae, an obligatory endophyte of pollen grains of Mediterranean pines.

Enterobacter cloacae was found to be associated with the pollen of several Mediterranean pines. The bacterium was detected only in mature pollen of Pinus halepensis, P. brutia, and P. pinea. E. cloacae is considered to be an obligatory endophyte based on its occurrence in disinfected male cones and the successful inoculation of seedlings of the above 3 species with E. cloacae AS1 isolated from pollen of P. halepensis used as a model strain. Strain AS1 was able to produce indolyl-3-acetic acid (IAA) from L-tryptophan in culture, and this was probably the source of the increased IAA content in the germination medium of pollen. In addition, strain AS1 promoted adventitious root formation in mung bean (Vigna radiata) cuttings. However, it was not possible to obtain bacterium-free pollen to elucidate its role in pollen germination.

Colony Count, Microbial↗
PubMed2004

Dendroremediation of trinitrotoluene (TNT). Part 1: Literature overview and research concept.

BACKGROUND, AIM AND SCOPE: For decades, very large areas of former military sites have been contaminated diffusely with the persistent nitroaromatic explosive 2,4,6-trinitrotoluene (TNT). The recalcitrance of the environmental hazard TNT is to a great extent due to its particulate soil existence, which leads to slow but continuous leaching processes. Although improper handling during the manufacture of TNT seems to be a problem of the past in developed countries, environmental deposition of TNT and other explosives is still going on unfortunately, resulting from thousands of unexploded ordnance or low order explosions at munitions test areas and at current battlefields. OBJECTIVE: Sustainable phytoremediation strategies for explosives in Germany, which intend to use trees to decontaminate soil and groundwater ('dendroremediation'), have to consider that most of the former German military sites are already covered with woodlands, mainly with conifer stands. Therefore, parallel investigation of the remediation potential is necessary for both of the selected hybrids of fast growing broadleaf trees, which are waiting for planting and forest conifers, which have already proven for decades that they are able to grow on explosive contaminated sites. MAIN FEATURES: A short literature review is given regarding phytoremediation of TNT with herbaceous plants and some general aspects of dendroremediation are discussed. Furthermore, an overview of our TNT-dendroremediation research network is introduced, which has the strategic goal to make dendroremediation more calculable for a series of potent trees for site-adapted in situ application and for the assessment of tree remediation potentials in natural attenuation processes. RESULTS AND DISCUSSION: Some of our methods, results and conclusions yet unpublished are presented. For a preliminary calculation of area-related annual TNT dendroremediation potential of five-year-old trees, the following values were assessed: Salix EW-13 6.0, Salix EW-20 8.5, Populus ZP-007 4.2, Betula pendula 5.2, Picea abies 1.9 and Pinus sylvestris 0.8 g m(-2) a(-1). For a 45-year-old spruce forest, an annual natural attenuation potential of 4.2 g TNT m(-2) a(-1) was found. CONCLUSION, RECOMMENDATIONS AND PERSPECTIVE: Our main results deliver quantitative proposals for dendroremediation strategies in situ and provide decision aids. Also aspects of growth of raw materials for energy production are considered. Our dendroremediation research concept for TNT and its congeners can be easily completed for other trees of interest and it can also be applied to herbaceous plants. Knowing the current bottlenecks of phytoremediation and considering the known environmental behaviour of other contaminants, elements of our methodological approach may be easily adapted to those pollutant groups, e.g. for pesticides, pharmaceuticals, PAHs, chlorinated recalcitrants and, with some restrictions, to inorganics and to multiple contaminations. Our dynamical dendrotolerance test systems will help to predict tree growth on polluted areas. To provide some light into the black box of TNT dendroremediation, experimental data regarding the uptake, distribution and degradation of [14C]-TNT in mature tree tissues will be reported in the second part of this publication.

Biodegradation, Environmental↗
PubMed2004

Avoidance of nonhost plants by a bark beetle, Pityogenes bidentatus, in a forest of odors.

The bark beetle, Pityogenes bidentatus (Coleoptera: Scolytidae), searches in mixed conifer and deciduous forests of northern Europe for suitable branches of its host, Scots pine ( Pinus sylvestris). We tested whether odors from several diverse nonhost trees and plants common in the habitat (e.g., mountain ash, Sorbus aucuparia; oak, Quercus robur; alder buckthorn, Frangula alnus; blueberry, Vaccinium myrtillus; raspberry, Rubus idaeus; and grass, Deschampsia flexuosa) would reduce the attraction of the bark beetle to traps releasing its aggregation pheromone components in the field. Volatiles from the leaves or bark of each of these plants significantly reduced the attraction of the beetles to their pheromone. Odors collected from these nonhosts and analyzed by GC/MS contained monoterpenes, sesquiterpenes, and "green-leaf" alcohols, several of which (e.g., 1-octene-3-ol and beta-caryophyllene) reduced the attraction to pheromone in the field and elicited electroantennographic responses. In the laboratory, reproduction by the beetle was marginal in nonhost Norway spruce, Picea abies, and was absent in the other nonhost trees. Olfactory avoidance of unsuitable nonhosts may have evolved due to advantages in avoiding mistakes during host selection.

Animals↗
PubMed2003

Testing for linkage disequilibrium in the New Zealand radiata pine breeding population.

Linkage analysis is commonly used to find marker-trait associations within the full-sib families of forest tree and other species. Study of marker-trait associations at the population level is termed linkage-disequilibrium (LD) mapping. A female-tester design comprising 200 full-sib families generated by crossing 40 pollen parents with five female parents was used to assess the relationship between the marker-allele frequency classes obtained from parental genotypes at SSR marker loci and the full-sib family performance (average predicted breeding value of two parents) in radiata pine ( Pinus radiata D. Don). For alleles (at a marker locus) that showed significant association, the copy number of that allele in the parents was significantly correlated, either positively or negatively, with the full-sib family performance for various economic traits. Regression of parental breeding value on its genotype at marker loci revealed that most of the markers that showed significant association with full-sib family performance were not significantly associated with the parental breeding values. This suggests that over-representation of the female parents in our sample of 200 full-sib families could have biased the process of detecting marker-trait associations. The evidence for the existence of marker-trait LD in the population studied is rather weak and would require further testing. The exact test for genotypic disequilibrium between pairs of linked or unlinked marker loci revealed non-significant LD. Observed genotypic frequencies at several marker loci were significantly different from the expected Hardy-Weinberg equilibrium. The possibilities of utilising marker-trait associations for early selection, among-family selection and selecting parents for the next generation of breeding are also discussed.

Breeding↗
PubMed2003

Identification of AFLP markers linked to a resistance gene against pine needle gall midge in Japanese black pine.

Bulked segregant and AFLP analyses of two mapping populations (R17 x S6 and R17 x S1) were used to identify markers linked to Rpgm, the only known gene responsible for resistance to pine needle gall midge in Pinus thunbergii Parl. Rpgm was found to be bracketed by ACCC/CCTTT(190) on one side at a distance of 6.6 cM and ACGT/CCCGC(250) at 15.3 cM on the other side. The segregation of these markers was analyzed in two other families in order to determine their phase and transferability. One of the two additional resistant parents carried ACCC/CCTTT(190) in the homozygous state while the marker was in coupling (plus marker allele linked with an R allele) in a resistant parent, R17. The marker ACGT/CCCGC(250) was in a repulsion phase in R17 and was not detected in the other two resistant pine trees. Out of four AFLP markers identified, only ACGT/CCAAT(290) was transferable in all resistant trees tested, although its phase was opposite for different trees. These results indicate that in applying those markers to select resistant trees, the phase state of the markers in each resistant tree with respect to Rpgm needs to be considered.

Animals↗
PubMed2004

Pine genes regulated by the necrotrophic pathogen Fusarium circinatum.

A targeted genomics approach was used to construct a cDNA array of potential pathogen-regulated genes for investigating host-pathogen interactions in pine trees (Pinus species). This array, containing a nonredundant set of 311 cDNAs, was assembled by combining smaller sets of cDNAs generated by differential display or suppression subtraction hybridization using a variety of pathogen treatments and elicitors. The array was probed to identify host genes regulated by Fusarium circinatum, a necrotrophic fungus that incited pitch canker disease on pine stems. A set of 29 cDNAs were induced during the disease state. Notably, cDNAs on the array that were derived from experiments with fusiform rust, incited by Cronartium quercuum f. sp. fusiforme (a biotrophic fungus) were unregulated by Fusarium. the results imply distinct genetic responses in pine to diseases incited by necrotrophs and biotrophs. This cDNA collection expands the genomics toolkit for understanding interactions between conifers and their microbial associates in forest ecosystems.

Base Sequence↗
PubMed2005

Genetic dissection of fusiform rust and pitch canker disease traits in loblolly pine.

Loblolly pine (Pinus taeda L.) exhibits genetic resistance to fusiform rust disease (incited by the biotrophic fungus, Cronartium quercuum f. sp. fusiforme) and pitch canker disease (incited by the necrotrophic fungus, Fusarium circinatum). In this study, a total of 14,015 loblolly pine cuttings from 1,065 clones were screened in controlled greenhouse conditions to identify phenotypes of clones, families, and parents that guide a genetic dissection of disease traits associated with pitch canker and fusiform rust. A total of 23,373 phenotypic data points were collected for lesion length (pitch canker) and gall score, gall length, and gall width (fusiform rust). We verified heritable fusiform rust and pitch canker traits and calculated parental, clonal, and full-sib family rankings for both diseases. Genetic correlations revealed that traits associated with fusiform rust are genetically distinct from one another, and that the genetic mechanisms underlying pitch canker and fusiform rust resistance are independent. The disease phenotyping described here is a critical step towards identifying specific loci and alleles associated with fusiform rust and pitch canker resistance.

Analysis of Variance↗