Procyanidins from grape pomace are suitable inhibitors of human endothelial NADPH oxidase

Procyanidins have been associated with a reduced risk of cardiovascular diseases such as atherosclerosis. However, the molecular mechanisms underlying this benefit are not fully understood. Increased reactive oxygen species (ROS) production generated by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is a common problem in different cardiovascular diseases. Our objective was to evaluate the effects of procyanidin-rich fractions from distilled grape pomace on NADPH oxidase activity in human umbilical vein endothelial cells (HUVEC). Three differently polymerized and galloylated procyanidin fractions were analyzed for their NADPH oxidase inhibitory activity in cell lysates and in HUVEC cultures. All of the three fractions, up to 1 μg/ml, equally inhibited isolated NADPH oxidase in HUVEC lysates in a concentration-dependent manner and independently of any superoxide anion scavenging activities. The procyanidin fractions even blocked NADPH oxidase activity in intact HUVEC, inhibiting ROS production at both extra- and intracellular levels. The fractions achieved the same effects that known NADPH oxidase inhibitors, such as diphenylene iodonium and apocynin, but they presented better hydrosolubility. Our results demonstrated that procyanidin from grape pomace inhibit human endothelial NADPH oxidase regardless of their polymerization degree and galloylation percentage. Therefore, procyanidins are suitable NADPH oxidase inhibitors which could serve as models for therapeutic alternatives for cardiovascular diseases.     

In vitro anti-bacterial and anti-adherence effects of natural polyphenolic compounds on oral bacteria

Aims: To investigate the action of different polyphenolic compounds, extracted from red wine, grape marc and pine bark, on oral bacteria. Methods and Results: The anti-microbial activity of extracts was examined by determining the Minimal Inhibitory Concentration and Minimal Bactericidal Concentration using the macro dilution broth technique. Their effect on the adherence was tested on growing cells of Streptococcus mutans on a glass surface and on a multi-species biofilm grown on saliva-coated hydroxyapatite discs. The effect on glucosyltransferase activity was analysed through the reductions in the overall reaction rate and the quantity of insoluble glucan (ISG) synthesized. Pine bark and grape marc extracts were the most effective inhibitors of the multi-species biofilm formation and of the ISG synthesis. Conclusion: The tested components showed an interesting anti-plaque activity in vitro. Significance and Impact of the Study: This is, to our knowledge, the first and the most complete report on the properties of wine and pine bark extracts that could be used for oral disease prevention purpose.     

樟子松乙醇提取物体外抗氧化活性研究

测定了樟子松球果、树皮、树叶、木质部4个部位提取物的总抗氧化能力、总还原力、二苯代苦味酰基（DPPH·）清除能力、羟基自由基（·OH）清除能力、超氧阴离子（O^·₂）清除能力5个指标,并以抗坏血酸（Vc）为阳性对照评价从樟子松4个部位提取的松多酚体外抗氧化活性。结果表明：（1）从樟子松树皮中提取的樟子松多酚表现出较强的抗氧化能力,在较高浓度下与抗氧化剂（Vc）相差较小;从樟子松球果中提取的樟子松多酚也表现出较强的还原力,甚至超过阳性对照抗氧化剂Vc;（2）从樟子松树皮中提取的樟子松多酚在二苯代苦味酰基（DPPH·）清除能力、羟基自由基（·OH）清除能力上均为三者之中最高的,但都不超过抗氧化剂（Vc）的抗氧化能;（3）超氧阴离子（O^·₂）清除能力。三者的清除率均为负值,说明该方法测定超氧阴离子（O^·₂）清除率存在干扰因素,无法比较三者在这一指标上的能力大小。上述结果表明樟子松乙醇提取物具有较强的抗氧化能力,可作为天然的抗氧化剂。     

Fractionation of grape tannins and analysis by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry

Polymeric tannins, extracted from grape berries (Gamay variety), were fractionated according to their mean degree of polymerisation (mDP) on a styrene-divinylbenzene phase eluted with a gradient of methanol:chloroform. Increasing the percentage of methanol led to the solubilisation of higher molecular weight tannins. The mean mDP of each collected fraction was determined by acid-catalysed degradation in the presence of a nucleophilic reagent. The fractionation method produced a linear gradient of mDP varying between 1.84 and 19.34. The fractions were partially characterised by matrix assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). The spectra showed a complex mixture of proanthocyanidins and galloylated proanthocyanidins up to 4000 amu.     

Survey and phylogenetic analysis of culturable microbes in the oral secretions of three bark beetle species

In a recent study, we reported a previously undescribed behavior in which a bark beetle exuded oral secretions containing bacteria that have antifungal properties, and hence defend their galleries against pervasive antagonistic Hyphomycete fungi. Actinobacteria, a group known for their antibiotic properties, were the most effective against fungi that invade the spruce beetle galleries. In the present study, we describe the isolation and identification of microorganisms from oral secretions of three bark beetles (Coleoptera: Curculionidae: Scolytinae): the spruce beetle, Dendroctonus rufipennis Kirby, the mountain pine beetle, Dendroctonus ponderosae Hopkins, and the pine engraver, Ips pini Say. Bacteria isolated from these three species span the major bacterial classes α-, β-, and γ-Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, except for D. ponderosae , which yielded no α-proteobacteria or Bacteroidetes isolates. Spruce beetles and pine engraver beetles had similar numbers of α-proteobacteria isolates, but pine engravers yielded twice as many Bacteroidetes isolates as spruce beetles. In contrast, mountain pine beetles yielded more isolates in the β- and γ-proteobacteria than spruce beetles and pine engravers. The highest percentage of Actinobacteria was obtained from spruce beetles, followed by pine engravers and mountain pine beetles. All of the fungal isolates obtained from the three beetle species were Ascomycetes. The greatest fungal diversity was obtained in spruce beetles, which had nine species, followed by pine engravers with five, and mountain pine beetles with one.     

Comparison between pine bark and coconut husk sorption capacity of metals and nitrate when mixed with sewage sludge

Waste products such as biosolids and wood wastes have been frequently used in container production of plants. The use of biosolids in agriculture is a beneficial mean of recycling the by-products of waste-water treatment. However, care must be taken to avoid environmental or human health problems via run-off and leaching. The objective of this work is to compare the retention capacity of cadmium, lead, zinc and nitrate between pine bark (PB) and coconut fibre (F) when mixed with increasing amounts of composted sewage sludge (CSS) (0%, 15% and 30% (v/v)). Substrates were packed into leaching columns and irrigated with deionised water every 2 days. Leachates were collected during 1 month, and nitrate, Zn, Cd, Pb, EC and pH were monitored along the experiment. PB columns leached lower amount of nitrate than the coconut fibre ones. The same trend was observed for Zn, Cd and Pb. It could be said, that in order to minimize the environmental risks of using sewage sludges our results indicate that it is preferred to mix the sludge with pine bark instead than with coconut husk.     

Reuse of waste materials as growing media for ornamental plants

The use of different waste materials: pine bark, coconut fibre and sewage sludge as substrates in the production of ornamental plants was studied, with an special interest on the suitability of coconut fibre as growing substrate for conifer plants. The plant species tested were Pinus pinea, Cupressus arizonica and C. sempervirens and the substrate mixtures were: (1) pine bark, (2) pine bark with 15% of sewage sludge compost, (3) pine bark with 30% of sewage sludge compost, (4) coconut fibre, (5) coconut fibre with 15% of sewage sludge compost and (6) coconut fibre with 30% of sewage sludge compost. Substrates were physically and chemically well characterized, and 75-cm plants were grown on them for one year. Plant and substrate status were periodically tested along the experiment. As biosolid recycling is the main objective of the present work, the mixtures with 30% of composted sewage sludge will be the most convenient substrate to use. For C. sempervirens and C. arizonica, a mixture between pine bark or coconut fibre and 30% of biosolid compost in volume gave the best results, but the lower cost of the pine bark than the coconut fibre substrate indicated the use of the PB+30% CSS. For P. pinea the research of new combinations between waste products is recommended to attain better results.     

Changes in populations of soil microorganisms,nematodes, and enzyme activity associated with application of powdered pine bark

Evaluation of enzyme activities in combination with taxonomic analyses may help define the mechanisms involved in microbial decomposition of orgaic amendments and biological control of soilborne pathogens. In this study, powdered pine bark was added to nematode-infested soil at rates of 0, 5, 10, 15, 20, 25, 30, 35, 40, 45, and 50 g kg^–1. Total fungal populations did not differ among treatments immediately after application of pine bark. After 7 days, fungal populations were positively correlated with increasing levels of pine bark. This increase was sustained through 14 and 21 days.Penicillium chrysogenum andPaecilomves variotii were the predominant fungal species isolated from soil amended with pine bark. Total bacterial populations did not change with addition of pine bark at 0, 7, and 14 days after treatment. At 21 and 63 days, total bacterial populations declined in soil receiving the highest rates of pine bark. Addition of pine bark powder to soil caused a shift in predominant bacterial genera fromBacillus spp. in nonamended soil, toPseudomonas spp. in amended soil. Soil enzyme activities were positively correlated with pine bark rate at all sampling times. Trehalase activity was positively correlated with total fungal populations and with predominant fungal species, but was not related to bacterial populations. The number of non-parasitic (non-stylet bearing) nematodes andMeloidogyne arenaria in soil and roots were not correlated with pine bark rate. However,Heterodera glycines juveniles in roots, and the number of cysts g^–1 root, declined with increasing levels of pine bark.Journal Series Series No. 18-933598 Alabama Agricultural Experiment Station     

Characterization of the photosynthetic apparatus in cortical bark chlorenchyma of Scots pine

Winter-induced inhibition of photosynthesis in Scots pine (Pinus sylvestris L.) needles is accompanied by a 65% reduction of the maximum photochemical efficiency of photosystem II (PSII), measured as F _v/F _m, but relatively stable photosystem I (PSI) activity. In contrast, the photochemical efficiency of PSII in bark chlorenchyma of Scots pine twigs was shown to be well preserved, while PSI capacity was severely decreased. Low-temperature (77 K) chlorophyll fluorescence measurements also revealed lower relative fluorescence intensity emitted from PSI in bark chlorenchyma compared to needles regardless of the growing season. Nondenaturating SDS-PAGE analysis of the chlorophyll–protein complexes also revealed much lower abundance of LHCI and the CPI band related to light harvesting and the core complex of PSI, respectively, in bark chlorenchyma. These changes were associated with a 38% reduction in the total amount of chlorophyll in the bark chlorenchyma relative to winter needles, but the Chl a/b ratio and carotenoid composition were similar in the two tissues. As distinct from winter pine needles exhibiting ATP/ADP ratio of 11.3, the total adenylate content in winter bark chlorenchyma was 2.5-fold higher and the estimated ATP/ADP ratio was 20.7. The photochemical efficiency of PSII in needles attached to the twig recovered significantly faster (28–30 h) then in detached needles. Fluorescence quenching analysis revealed a high reduction state of Q _A and the PQ-pool in the green bark tissue. The role of bark chlorenchyma and its photochemical performance during the recovery of photosynthesis from winter stress in Scots pine is discussed.     

Inhibition of advanced glycation by flavonoids. A nutritional implication for preventing diabetes complications?

Advanced glycation of collagens contributes to development of micro- and macrovascular complications in diabetes. Since flavonoids are potent natural antioxidants, it was interesting to examine their effect on the formation of a cross-linking advanced glycation endproduct, pentosidine, in collagen incubated with glucose. Monomeric flavonoids (25 and 250 microM) markedly reduced pentosidine/hydroxyproline values in a concentration- and structure-dependent manner. Procyanidin oligomers from grape seed were more active than pine bark procyanidin oligomers. Oligomers are known to be cleaved into monomers in the gastric milieu and monomeric flavonoids to be absorbed and recovered at micromolar concentrations (with a long plasmatic half-life) in extracellular fluids, in contact with collagens. In conclusion, flavonoids are very potent inhibitors of pentosidine formation in collagens, active at micromolar concentrations; these concentrations might be achieved in plasma of diabetic patients after oral intake of flavonoids.     

凉水自然保护区不同皮型红松径向生长对气候的响应

运用相关函数及单年分析等树木年轮气候学方法,研究了黑龙江凉水国家自然保护区不同皮型红松径向生长与气候因子的关系、主要影响因子及这种响应关系是否长期稳定.结果表明: 细皮红松更适合做树木年轮气候学分析.两种皮型红松的径向生长对环境变化都比较敏感,对气候因子的响应无显著差异.1902—2009年生长季,尤其当年6月的气候因子是影响研究区两种皮型红松生长的主要因子.其中,温度表现为显著负相关,降水表现为显著正相关.不同时间段内红松径向生长对气候因子的响应存在差异,随着1970年后气温的快速升高及干旱的加剧,两种皮型红松径向生长对气候因子的响应更敏感,尤其表现为对生长季温度和更多季节水热复合因子变化（帕尔默干旱指数）的响应更显著.
     

Evaluation of powdered pine bark for control ofMeloidogyne arenaria andHeterodera glycines on soybean

Commercially avaiable pine bark nuggets (composted) and fresh pine bark were ground into powders and added at rates of 0 to 50 g kg^–1 to field soil infested withMeloidogyne arenaria andHeterodera glycines. The treated soil was maintained moist in the greenhouse for 2 weeks, sampled, and planted with Davis soybean (Glycine max.). Eight weeks after planting, numbers ofM. arenaria andH. glycines in soil decreased with increasing amounts of composted or fresh pine bark. No juveniles were present in soil treated with 5% pine bark. The number of galls and cysts g^–1 root decreased in proportion to the amount of pine bark added to soil. Gall and cyst formation was completely eliminated at the 5% rate. Numbers of saprophagous nematodes were highest in soil with 4–5% pine bark. The activity of several soil enzymes was correlated with the addition of both composted and fresh pine bark. Fresh pine bark powder caused an increase in soil enzyme activity compared to composted pine bark, but did not provide consistent control of gall and cyst formation.Journal Series No. 18-933579 Alabama Agricultural Experiment Station     

Anthropogenic fire and bark thickness in coastal and island pine populations from Alta and Baja California

Aim  Native American fire use influenced bark thickness of coastal and island Monterey pine ( Pinus radiata D. Don) and bishop pine ( Pinus muricata D. Don) populations.
Methods  Basal bark thickness and d.b.h. were measured in two common-garden pine plantations that included all five native Monterey pine populations and nine of 10 native bishop pine populations. One-factor analysis of covariance was used to determine if significant differences in bark thickness existed between island and coastal populations.
Results  Bark thickness was measured on 228 Monterey and 42 bishop pines. Bark thickness in coastal and island Monterey pine populations was significantly different. Bark thickness in coastal and island bishop pine populations was not significantly different.
Main conclusions  Basal bark was thick in populations where there was a history of Native American burning. Basal bark was thin in two island populations where Native Americans have been absent or distant from the pine populations. While other influences no doubt affect the evolution of lower-bole bark thickness, it appears that frequent anthropogenic fires may be a powerful selection force.     

Nutritional disturbances of young Scots pines caused by pine bark bugs in a dry heath forest

This study deals with the effect of pine bark bugs (Aradus cinnamomeus Panzer) on the nutrition of young Scots pines (Pinus sylvestris L.). Soil and needle samples for analytical purposes were collected from a young pine stand growing on a dry barren mineral soil afflicted by pine bark bugs.The damage to vascular tissues caused by pine bark bugs disturbed the nutrition of the trees, especially in the top part of the crown. The foliar calcium, magnesium, manganese and sulphur concentrations were highly reduced. Scots pines suffered from a lack of calcium, magnesium, nitrogen and phosphorus. These deficiencies were secondary by nature, because no differences were observed between the nutrient concentrations of the underlying soil of the healthy and affected trees. The symptoms of trees damaged by pine bark bugs resembled most of all calcium deficiency symptoms.     

The microbiology of pine bark composting: An Electron-microscope and physiological study

An electron microscope study was conducted on samples of pine bark taken from stacks during consecutive stages of composting. It was found using scanning electron microscopy (SEM) that bacteria, actinomycetes and fungi were present in relatively low numbers on the bark surface before composting was initiated. After addition of urea and water to bark heaps, microbial numbers rose, particularly the bacterial fraction. A large number of actinomycetes were seen below the surface of the bark as composting progressed. Transmission electron microscopy (TEM) of bark in the late stages of composting demonstrated the presence of a variety of microbes within the bark cells. The microorganisms were seen, using SEM, to be degrading the surface of the bark chips, and, using TEM, to be attached to the lignified cell walls. Physiological studies on bacteria isolated at different stages of composting showed they had a number of enzymes such as carboxymethyl cellulase that could aid in the degradation of pine bark. The isolates consisted of Gram-negative and Gram-positive strains, some of which were spore formers. Most of the isolates, including some Gram-negative non-sporing bacteria, were able to grow over a wide range of temperatures from 30 to 60°C, and, in some cases, 70°C.     

Fractionation of Forest Residues of Douglas-fir for Fermentable Sugar Production by SPORL Pretreatment

Douglas-fir (Pseudotsuga menziesii) forest residues were physically fractionated through sieving. The bark and wood were separated for large-sized fractions (>12.7?mm), and their contents were determined. The chemical compositions of the large fractions were calculated based on the contents and chemical compositions of the bark and wood. The chemical compositions of the fine fractions were analyzed. The bark and wood content in the fine fractions was calculated based on the measured glucan and lignin contents in each fraction. It was found that fractionation by particle/chip size can effectively fractionate bark and wood and therefore lignin from carbohydrates. The large-sized fractions (>12.7?mm) represent approximately 60?% of the collected forest residues but only contain approximately 37?% of the total bark and 35?% of the total ash, or a selectivity over bark and ash of 1.6 and 1.7, respectively. Pretreatment of forest residues by sulfite pretreatment to overcome recalcitrance of lignocelluloses and subsequent enzymatic hydrolysis revealed the presence of 14.3?% bark can reduce substrate enzymatic digestibilities (SED) 16?% compared with that from a bark-free sample. The SED of a bark is 41?% compared with 73?% for wood when pretreated under the same conditions. Separating pretreatment of bark from wood is beneficial for producing a more enzymatically digestible substrate. The results from the present study could have significant implications for harvesting forest residues.     

Activity of grape extracts from Greek varieties of Vitis vinifera against mutagenicity induced by bleomycin and hydrogen peroxide in Salmonella typhimurium strain TA102

Several in vivo and in vitro studies have shown that grape extracts could prevent certain steps in carcinogenesis and a few mechanisms have been proposed for this activity. In this study, the potential antimutagenic activity of methanolic and aqueous extracts from two Greek grape varieties of Vitis vinifera against DNA damage induced by reactive oxygen species (ROS) was assessed as a potential novel chemopreventive mechanism, using Salmonella typhimurium strain TA102. The two grape varieties were Assyrtiko (white grapes) and Mandilaria (red grapes), while the oxidant mutagens used were bleomycin (BLM) and hydrogen peroxide (H(2)O(2)). Since it has been considered that polyphenols present in grapes are their most potent biologically active compounds, we also tested the effects of polyphenol-rich fractions as well as some of the more common grape polyphenols on the activity of the two test mutagens. These polyphenols were quercetin, (+)-catechin, (-)-epicatechin, trans-resveratrol, gallic acid and protocatechuic acid. Almost all extracts showed inhibitory activity against both mutagens. On the other hand, polyphenol-rich fractions as well as individual polyphenols at concentrations found in the extracts either did not diminish or did enhance the activity of the mutagens. These results suggest that the protection of DNA from mutations induced by ROS may be one of the mechanisms accounting for the chemopreventive activity of grape extracts. However, it seems that this protective activity may not be attributed to polyphenols but rather to a synergism of many compounds in the grapes.     

Seasonal fluctuation of dehydrins is related to osmotic status in Scots pine needles

Seasonal variation in dehydrins and other soluble proteins of Scots pine (Pinus sylvestris L.) needles, buds and bark were analyzed monthly for 1 year from 1998 to 1999. Dehydrin-related proteins of 60 and 56 kDa were identified immunologically in all tissues. The concentration of the 60-kDa dehydrin was highest during the winter (October-February) in buds and bark but increased in early spring (March-May) in needles. Accumulation of the 60-kDa dehydrin in the needles in springtime was related to the decreasing osmotic potentials of the needles. The 56-kDa dehydrin was present only during the growing season, as was a 50-kDa dehydrin, which only appeared in bud and bark tissues. The soluble protein concentration of needles did not differ significantly between seasons, but in bark and bud tissues the protein concentrations were at their lowest level in newly grown tissues (June-August). The level of several polypeptides was higher during the winter-spring period than in the growing season, especially in bark and bud tissues. These proteins may be related to cold hardiness or dormancy in overwintering Scots pine. Dehydrin-related proteins in needles are linked to springtime changes in the osmotic status of needles rather than to their cold acclimation.     

Study of biochemical and microbiological parameters during composting of pine and eucalyptus bark

To study the possibility of some residues from pulp and paper industry being used as substrates to produce seedlings in containers, three composting experiments were carried out using eucalyptus bark, pine bark and a mixture (60:40, v:v) of pine bark+eucalyptus bark. Biochemical parameters studied were: acid and alkaline phosphatases, lipase (C10), protease, urease, beta-glucosidase and total cellulases. The microbiological populations of total aerobic bacteria, total fungi, actinomycetes, nitrifying bacteria, cellulolytic bacteria and fungi were also evaluated. At the end of the process physicochemical characterization of composts was also performed. Results showed in general that the highest microbiological populations as well as for enzymatic activities occurred during the thermophilic phase (>40 degrees C) of the process. On the other hand and according to the physicochemical characteristics of composts pine bark is the most appropriate material to be used in the formulation of substrates to produce plants in containers.