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1.  High genetic diversity with moderate differentiation in Juniperus excelsa from Lebanon and the eastern Mediterranean region 
AoB Plants  2011;2011:plr003.
Juniperus excelsa constitutes a precious woody species of high ecological value able to grow up to Mountain treeline around the Mediterranean. Nuclear microsatellites were used to shed light on genetic diversity and differentiation of different Mediterranean populations. This information is essential in planning conservation strategies and reforestation programs.
Background and aims
Juniperus excelsa is an important woody species in the high mountain ecosystems of the eastern Mediterranean Basin where it constitutes the only coniferous species found at the tree line. The genetic diversity within and among J. excelsa populations of the eastern Mediterranean Basin is studied in the light of their historical fragmentation.
Methodology
Nuclear microsatellites originally developed for Juniperus communis and J. przewalskii were tested on 320 individuals from 12 different populations originating from Lebanon, Turkey, Cyprus, Greece and the Ukraine.
Principal results
Among the 31 nuclear microsatellite primers tested, only three produced specific amplification products, with orthology confirmed by sequence analysis. They were then used for genetic diversity studies. The mean number of alleles and the expected heterozygosity means were Na=8.78 and He=0.76, respectively. The fixation index showed a significant deviation from Hardy–Weinberg equilibrium and an excess of homozygotes (FIS=0.27–0.56). A moderate level of genetic differentiation was observed among the populations (FST=0.075, P<0.001). The most differentiated populations corresponded to old vestigial stands found at the tree line (>2000 m) in Lebanon. These populations were differentiated from the other populations that are grouped into three sub-clusters.
Conclusions
High levels of genetic diversity were observed at species and population levels. The high level of differentiation in the high-mountain Lebanese populations reflects a long period of isolation or possibly a different origin. The admixture observed in other populations from Lebanon suggests a more recent separation from the Turkish–southeastern European populations.
doi:10.1093/aobpla/plr003
PMCID: PMC3064508  PMID: 22476474
2.  Antioxidant Activity of the Essential Oils of Different Parts of Juniperus excelsa M. Bieb. subsp. excelsa and J. excelsa M. Bieb. subsp. polycarpos (K. Koch) Takhtajan (Cupressaceae) 
The essential oils of branchlets and fruits of Juniperus excelsa subsp. excelsa and Juniperus excelsa subsp. polycarpos were examined for their antioxidant activity. The compositions of the essential oils were studied by GC and GC-MS. To evaluation the antioxidants activity of the volatile oils, pure components and positive controls at different concentrations, thin-layer chromatography (TLC) screening methods, diphenylpicrylhydrazyl (DPPH) assay, deoxyribose degradation test and modified deoxyribose degradation test were employed. The results of the present study demonstrate some antioxidant activity for the tested essential oils obtained from various parts of both plants. It indicates that the use of these essential oils, in very low concentrations, may be useful as a natural preservative. However before any final conclusion, it is suggested that the antioxidant activity of these oils should also be evaluated by using lipid solvent system methods.
PMCID: PMC3813057  PMID: 24250416
Juniperus excelsa subsp. Excels; Juniperus excelsa subsp. Polycarpos; Cupressaceae; Essential oils; Antioxidant activity
3.  In-vitro Antimicrobial Activities of Some Iranian Conifers 
Male and female leaves and fruits of eleven different taxons of Iranian conifers (Cupressus sempervirens var. horizontalis, C. sempervirens var. sempervirens, C. sempervirens cv. Cereifeormis, Juniperus communis subsp. hemisphaerica, J. excelsa subsp. excelsa, J. excelsa subsp. polycarpos, J. foetidissima, J. oblonga, J. sabina, Platycladus orientalis and Taxus baccata) were collected from different localities of Iran, dried and extracted with methanol. The extracts were tested for their antimicrobial activity against Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and Candida albicans. The extracts were screened qualitatively using four different methods, the disc diffusion, hole plate, cylinder agar diffusion and agar dilution methods, whereas the minimum inhibitory concentrations (MIC) of each extract were determined by the agar dilution method. The best result was obtained by means of hole plate method in qualitative determination of antimicrobial activities of extracts and the greatest activity was found against S. aureus in all tested methods.
PMCID: PMC3813213  PMID: 24250573
Antimicrobial activity; Cupressus; Cupressaceae; Juniperus; Platycladus; Taxaceae; Taxus
4.  Antioxidant Activity of Leaves and Fruits of Iranian Conifers 
Cupressus semipervirens var. horizontalis, Cupressus semipervirens var. semipervirens, Cupressus semipervirens cv. Cereifeormis, Juniperus communis subsp. hemisphaerica, Juniperus excelsa subsp. excelsa, Juniperus excelsa subsp. polycarpos, Juniperus foetidissima, Juniperus oblonga, Juniperus sabina, Platycladus orientalis and Taxus baccata are Iranian conifers. The antioxidant activity of leaves and fruits of these 11 different taxons were evaluated. The leaves of both male and female, and fruits of these plants were collected from different areas of the country. Methanol extract of leaves and fruits of these taxons were prepared. Antioxidant activity of each extracts was measured using two different tests of the ferric thiocyanate method and thiobarbituric acid. Results indicated that the methanol extracts of leaves, of male and female, and fruits of all these species (27 samples) possessed antioxidant activity when tested with both methods. The antioxidant activity was then compared with those of α-tocopherol (a natural antioxidant) and butylated hydroxytoluene (a synthetic antioxidant). Methanol extract of fruits of C. semipervirens cv. Cereifeormis showed the highest antioxidant activity while the methanol extract of leaves of C. semipervirens var. semipervirens possessed the lowest antioxidant activity. However, our finding showed that most of the tested extracts were showing strong antioxidant activity even higher than α-tocopherol.
doi:10.1093/ecam/nem011
PMCID: PMC1978238  PMID: 17965761
antioxidative activity; conifers, Cupressus; ferric thiocyanate test; Juniperus; medicinal plant; Platycladus orientalis; radical scavenging; thiobarbituric acid test
5.  Analysis of antimicrobial, antifungal and antioxidant activities of Juniperus excelsa M. B subsp. Polycarpos (K. Koch) Takhtajan essential oil 
Pharmacognosy Research  2010;2(3):128-131.
Juniperus excelsa M.B subsp. Polycarpos (K.Koch), collected from south of Iran, was subjected to hydrodistillation using clevenger apparatus to obtain essential oil. The essential was analyzed by gas chromatography/mass spectrometry (GC/MS) and studied for antimicrobial, antifungal and antioxidant activities. The results indicated α-pinene (67.71%) as the major compound and α-cedral (11.5%), δ3-carene (5.19%) and limonene (4.41%) in moderate amounts. Antimicrobial tests were carried out using disk diffusion method, followed by the measurement of minimum inhibitory concentration (MIC). All the Gram positive and Gram negative bacteria were susceptible to essential oil. The oil showed radical scavenging and antioxidant effects.
doi:10.4103/0974-8490.65505
PMCID: PMC3141302  PMID: 21808554
Antimicrobial activity; antioxidant activity; essential oil; Juniperus excelsa; thin layer chromatography autographic assay
6.  Purification, crystallization and preliminary crystallographic analysis of peroxidase from the palm tree Chamaerops excelsa  
Diffraction-quality crystals of the peroxidase from the palm tree C. excelsa were obtained and a native X-ray diffraction data set was collected at a synchrotron source.
Plant peroxidases are presently used extensively in a wide range of bio­technological applications owing to their high environmental and thermal stability. As part of efforts towards the discovery of appealing new biotechnological enzymes, the peroxidase from leaves of the palm tree Chamaerops excelsa (CEP) was extracted, purified and crystallized in its native form. An X-­ray diffraction data set was collected at a synchrotron source and data analysis showed that the CEP crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 70.2, b = 100.7, c = 132.3 Å.
doi:10.1107/S1744309111039030
PMCID: PMC3232160  PMID: 22139187
palm trees; peroxidases; Chamaerops excelsa; extremoenzymes
7.  Taxonomic Study of Amanita Subgenus Lepidella and Three Unrecorded Amanita Species in Korea 
Mycobiology  2013;41(4):183-190.
Amanita Pers. is a well-known monophyletic mushroom genus with a broad distribution. However, the diversity of Korean Amanita species has been underestimated, and most taxonomic studies conducted in Korea have only investigated their morphological characteristics. This approach is frequently insufficient for correct identification in fungal classification; therefore, we constructed a phylogeny of Amanita subgen. Lepidella in order to understand the phylogenetic placements of 16 Amanita specimens collected in Korea in 2012. The phylogeny constructed using the sequence data of the internal transcribed spacers and the partial large subunit of ribosomal RNA identified nine Amanita species (A. citrina, A. excelsa var. spissa, A. flavipes, A. fritillaria, A. oberwinklerana, A. pallidorosea, A. rubescens, A. subjunquillea, and A. volvata); of these, A. fritillaria, A. oberwinklerana, and A. pallidorosea are new to Korea.
doi:10.5941/MYCO.2013.41.4.183
PMCID: PMC3905121  PMID: 24493938
Amanita subgen. Lepidella; A. fritillaria; A. oberwinklerana; A. pallidorosea; Morphology; Phylogeny
8.  Purification, crystallization and initial crystallographic characterization of brazil-nut allergen Ber e 2 
The crystallization of the brazil nut allergen Ber e 2 is reported.
Peanut and tree-nut allergies have attracted considerable attention because of their frequency and their lifelong persistence. Brazil-nut (Bertholletia excelsa) allergies have been well documented and the 11S legumin-like seed storage protein Ber e 2 (excelsin) is one of the two known brazil-nut allergens. In this study, Ber e 2 was extracted from brazil-nut kernels and purified to high purity by crystalline precipitation and gel-filtration chromatography. Well diffracting single crystals were obtained using the hanging-drop vapour-diffusion method. A molecular-replacement structural solution has been obtained. Refinement of the structure is currently under way.
doi:10.1107/S1744309107051445
PMCID: PMC2339761  PMID: 18007055
brazil nut allergy; Ber e 2; cupin superfamily; allergenicity
9.  Effect of Oxygen-Reducing Atmospheres on the Safety of Packaged Shelled Brazil Nuts during Storage 
This work reports the application of oxygen-(O2-) reducing atmosphere methods on stored shelled Brazil nut (Bertholletia excelsa H.B.K.) packs aiming to evaluate the degree of aflatoxin degradation, nuts lipid oxidative stability, fungi control, and hygienic conditions improvement. The methods applied were (a) ozone: O3, (b) carbon dioxide: CO2, and (c) O2 absorber pads with and without vacuum. From all modified atmospheres evaluated, the best performance was obtained with O3, either with or without vacuum. It was the only nut treatment that was able to degrade aflatoxins. None of the spiked (AFLs: 15 μg·kg−1) nut samples O3- treated had aflatoxins detected up to the LC-MS/MS method LOQ (0.36 μg·kg−1 for total AFLs), thus producing safer nuts. Also it kept the fatty acid oxidation indicator—malondialdehyde stable and improved the sensory attributes for consumer acceptance. In addition, the destruction of fungi and yeast was observed since the O3 application (from 1.8 × 104 cfu/g to NG = no growth). All other treatments stabilized and/or inhibited microorganisms' growth only. By adding CO2 gas also played an important role in the nut quality. Regarding cost, gaseous O3 showed to be of low cost for application in the nut packs.
doi:10.1155/2011/813591
PMCID: PMC3132530  PMID: 21760791
10.  Aspergillus bertholletius sp. nov. from Brazil Nuts 
PLoS ONE  2012;7(8):e42480.
During a study on the mycobiota of brazil nuts (Bertholletia excelsa) in Brazil, a new Aspergillus species, A. bertholletius, was found, and is described here. A polyphasic approach was applied using morphological characters, extrolite data as well as partial β-tubulin, calmodulin and ITS sequences to characterize this taxon. A. bertholletius is represented by nineteen isolates from samples of brazil nuts at various stages of production and soil close to Bertholletia excelsa trees. The following extrolites were produced by this species: aflavinin, cyclopiazonic acid, kojic acid, tenuazonic acid and ustilaginoidin C. Phylogenetic analysis using partial β-tubulin and camodulin gene sequences showed that A. bertholletius represents a new phylogenetic clade in Aspergillus section Flavi. The type strain of A. bertholletius is CCT 7615 ( = ITAL 270/06 = IBT 29228).
doi:10.1371/journal.pone.0042480
PMCID: PMC3428331  PMID: 22952594
11.  Screening of Antibacterial Potentials of Some Medicinal Plants from Melghat Forest in India 
Cyperus rotundus, Caesalpinia bonducella, Tinospora cordifolia, Gardenia gummifera, Ailanthus excelsa, Acacia arabica, Embelia ribes and Ventilago maderspatana from Melghat forest were screened for their antibacterial potential against Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Proteus vulgaris, Salmonella typhi, Shigella flexneri, Salmonella paratyphi, Salmonella typhimurium, Pseudomonas aeruginosa, Enterobacter aerogenes by disc diffusion method. Out of these medicinal plants Caesalpinia bonducella, Gardenia gummifera and Acacia arabica showed remarkable antibacterial potential. The phytochemical analysis had showed the presence of Cardiac glycosides in all extracts (aqueous, acetone, ethanol and methanol) of Acacia arabica, Gardenia gummifera and ethanol, methanol extracts of Caesalpinia bonducella. Flavonoids were present in Gardenia gummifera, Ailanthus excelsa and acetone, methanol extracts of Acacia Arabica. Tannins and phenolic were present in Cyperus rotundus, Embelia ribes, and organic extracts of Ventilago maderspatana.
PMCID: PMC2816464  PMID: 20448847
Antibacterial activity; Melghat; Medicinal Plants; Phytochemical
12.  A Novel Triterpenoid Isolated from the Root Bark of Ailanthus excelsa Roxb (Tree of Heaven), AECHL-1 as a Potential Anti-Cancer Agent 
PLoS ONE  2009;4(4):e5365.
Background
We report here the isolation and characterization of a new compound Ailanthus excelsa chloroform extract-1 (AECHL-1) (C29H36O10; molecular weight 543.8) from the root bark of Ailanthus excelsa Roxb. The compound possesses anti-cancer activity against a variety of cancer cell lines of different origin.
Principal Findings
AECHL-1 treatment for 12 to 48 hr inhibited cell proliferation and induced death in B16F10, MDA-MB-231, MCF-7, and PC3 cells with minimum growth inhibition in normal HEK 293. The antitumor effect of AECHL-1 was comparable with that of the conventional antitumor drugs paclitaxel and cisplatin. AECHL-1-induced growth inhibition was associated with S/G2-M arrests in MDA-MB-231, MCF-7, and PC3 cells and a G1 arrest in B16F10 cells. We observed microtubule disruption in MCF-7 cells treated with AECHL-1 in vitro. Compared with control, subcutaneous injection of AECHL-1 to the sites of tumor of mouse melanoma B16F10 implanted in C57BL/6 mice and human breast cancer MCF-7 cells in athymic nude mice resulted in significant decrease in tumor volume. In B16F10 tumors, AECHL-1 at 50 µg/mouse/day dose for 15 days resulted in increased expression of tumor suppressor proteins P53/p21, reduction in the expression of the oncogene c-Myc, and downregulation of cyclin D1 and cdk4. Additionally, AECHL-1 treatment resulted in the phosphorylation of p53 at serine 15 in B16F10 tumors, which seems to exhibit p53-dependent growth inhibitory responses.
Conclusions
The present data demonstrate the activity of a triterpenoid AECHL-1 which possess a broad spectrum of activity against cancer cells. We propose here that AECHL-1 is a futuristic anti-cancer drug whose therapeutic potential needs to be widely explored for chemotherapy against cancer.
doi:10.1371/journal.pone.0005365
PMCID: PMC2671403  PMID: 19399188
13.  Micropropagation of Araucaria excelsa R. Br. var. glauca Carrière from orthotropic stem explants 
The objectives of the present work were in vitro propagation of Araucaria excelsa R. Br. var. glauca Carrière (Norfolk Island pine) with focus on the evaluation of the mean number of shoots per explant (MNS/E) and mean length of shoots per explants (MLS/E) produced by different parts of the orthotropic stem of A. excelsa R. Br. var. glauca in response to plant growth regulators. Norfolk Island pine axillary meristems responded very well to the 2-iso-pentenyl adenine (2iP) and thidiazuron (TDZ) levels. Explants taken from stem upper segments in the media containing 2iP had a higher MNS/E (3.47) and MLS/E (6.27 mm) in comparison to those taken from stem lower segments, which were 0.71 and 0.51 mm, respectively. Using 0.045 μM TDZ in the MS medium not only resulted in 4.60 MNS/E with 7.08 mm MLS/E but proliferated shoots showed a good performance as well. Investigating the best position of stem explant on mother plant as well as the best concentrations of growth regulators were performed which were useful for efficient micropropagation of this plant. Thirty three percent of explants were rooted in the MS medium containing 3 % sucrose, supplemented with 7.5 μM of both NAA and IBA for 2 weeks before transferring to a half strength MS medium without any growth regulator. Plantlets obtained were acclimatized and transferred to the greenhouse with less than 20 % mortality. This procedure considered the first successful report for regeneration and acclimatization of A. excelsa R. Br. var. glauca plantlet through main stem explants.
doi:10.1007/s12298-012-0115-9
PMCID: PMC3550516  PMID: 23814441
Abnormality; Araucaria excelsa R. Br. var. glauca; Axillary meristems; Orthotropic stems; Proliferation; Topophysis
14.  Ecosystem Consequences of Tree Monodominance for Nitrogen Cycling in Lowland Tropical Forest 
PLoS ONE  2013;8(7):e70491.
Understanding how plant functional traits shape nutrient limitation and cycling on land is a major challenge in ecology. This is especially true for lowland forest ecosystems of the tropics which can be taxonomically and functionally diverse and rich in bioavailable nitrogen (N). In many tropical regions, however, diverse forests occur side-by-side with monodominant forest (one species >60% of canopy); the long-term biogeochemical consequences of tree monodominance are unclear. Particularly uncertain is whether the monodominant plant-soil system modifies nutrient balance at the ecosystem level. Here, we use chemical and stable isotope techniques to examine N cycling in old-growth Mora excelsa and diverse watershed rainforests on the island of Trinidad. Across 26 small watershed forests and 4 years, we show that Mora monodominance reduces bioavailable nitrate in the plant-soil system to exceedingly low levels which, in turn, results in small hydrologic and gaseous N losses at the watershed-level relative to adjacent N-rich diverse forests. Bioavailable N in soils and streams remained low and remarkably stable through time in Mora forests; N levels in diverse forests, on the other hand, showed high sensitivity to seasonal and inter-annual rainfall variation. Total mineral N losses from diverse forests exceeded inputs from atmospheric deposition, consistent with N saturation, while losses from Mora forests did not, suggesting N limitation. Our measures suggest that this difference cannot be explained by environmental factors but instead by low internal production and efficient retention of bioavailable N in the Mora plant-soil system. These results demonstrate ecosystem-level consequences of a tree species on the N cycle opposite to cases where trees enhance ecosystem N supply via N2 fixation and suggest that, over time, Mora monodominance may generate progressive N draw-down in the plant-soil system.
doi:10.1371/journal.pone.0070491
PMCID: PMC3723728  PMID: 23936215
15.  Gibberella xylarioides Sensu Lato from Coffea canephora: a New Mating Population in the Gibberella fujikuroi Species Complex 
Applied and Environmental Microbiology  2005;71(12):8466-8471.
Gibberella xylarioides Heim & Saccas (presumed anamorph, Fusarium xylarioides Steyaert) is the causal agent of coffee wilt disease, an economically important tracheomycosis in Africa. In vitro crosses carried out with Congolese, Ugandan, and Tanzanian single-ascospore/conidial isolates originating from diseased Coffea canephora/excelsa demonstrated a heterothallic mating system, controlled by a single locus with two alleles, MAT-1 and MAT-2. Compatible isolates produced fertile perithecia within 2 to 8 weeks after mating. Mating type (MAT) was characterized by PCR with primer pairs previously developed for the Gibberella fujikuroi species complex (GFC) and for Fusarium oxysporum. All strains analyzed were morphologically identical and corresponded to Booth's description of the “female” F. xylarioides strain. Based on crossing results and MAT-2/translation elongation 1-α (tef) sequence data, G. xylarioides, as currently understood, is demonstrated to encompass at least three “groups”: G. xylarioides sensu strictu Ia, defined hitherto by two “historical” West African strains originating from the severe 1930s to 1950s epidemic (CBS 25852 and CBS 74979); G. xylarioides sensu strictu Ib, defined by two “historical” Central African lowland strains (DSMZ 62457 and ATCC 15664); and G. xylarioides sensu lato II, containing Congolese, Ugandan, and Tanzanian C. canephora/excelsa isolates. Infertility of crosses between the coffee wilt pathogen and known GFC mating populations demonstrates that G. xylarioides sensu lato constitutes a new biological species within the G. fujikuroi complex. MUCL 44532/MUCL 43887 and MUCL 35223/MUCL 44549 are proposed as G. xylarioides sensu lato II MAT-1/MAT-2 reference mating type tester strains.
doi:10.1128/AEM.71.12.8466-8471.2005
PMCID: PMC1317314  PMID: 16332836
16.  Genetic Variation in Five Mediterranean Populations of Juniperus phoenicea as Revealed by Inter-Simple Sequence Repeat (ISSR) Markers 
Annals of Botany  2006;97(2):299-304.
• Background and Aims The assessment of the genetic variability and the identification of isolated populations within a given species represent important information to plan conservation strategies on a genetic basis. In this work, the genetic variability in five natural populations of Juniperus phoenicea, three from Sardinia, one from Cyprus and the last one in the Maritime Alps was analysed by means of ISSRs, on the hypothesis that the latter could have been a refugial one during the last glaciation.
• Methods ISSRs were chosen because of their ability to detect variation without any prior sequence information. The use of three primers yielded 45 reproducible, polymorphic bands, which were utilized to estimate the basic parameters of genetic variability and diversity.
• Key Results All of the populations analysed harboured an adequate amount of genetic variability, with HS = 0·1299. The proportion of genetic diversity between populations has been estimated by GST = 0·12. The three Sardinian populations are separated, as tested by AMOVA, from the Cyprus and the continental ones.
• Conclusions The results indicate that geographical isolation has represented a major barrier to gene flow in Juniperus phoenicea. This work represents a first step towards a full genetic characterization of a conifer from the Mediterranean, a world biodiversity hotspot confronted with climate change, and thus contributes towards the planning of genetics-informed conservation strategies.
doi:10.1093/aob/mcj024
PMCID: PMC2803359  PMID: 16311272
Juniperus phoenicea L; genetic variation; ISSR; conservation; Cupressaceae
17.  Genetic rescue of remnant tropical trees by an alien pollinator. 
Habitat fragmentation is thought to lower the viability of tropical trees by disrupting their mutualisms with native pollinators. However, in this study, Dinizia excelsa (Fabaceae), a canopy-emergent tree, was found to thrive in Amazonian pastures and forest fragments even in the absence of native pollinators. Canopy observations indicated that African honeybees (Apis mellifera scutellata) were the predominant floral visitors in fragmented habitats and replaced native insects in isolated pasture trees. Trees in habitat fragments produced, on average, over three times as many seeds as trees in continuous forest, and microsatellite assays of seed arrays showed that genetic diversity was maintained across habitats. A paternity analysis further revealed gene flow over as much as 3.2 km of pasture, the most distant pollination precisely recorded for any plant species. Usually considered only as dangerous exotics, African honeybees have become important pollinators in degraded tropical forests, and may alter the genetic structure of remnant populations through frequent long-distance gene flow.
doi:10.1098/rspb.2001.1781
PMCID: PMC1088891  PMID: 11703880
18.  Brazil nuts intake improves lipid profile, oxidative stress and microvascular function in obese adolescents: a randomized controlled trial 
Background
Obesity is a chronic disease associated to an inflammatory process resulting in oxidative stress that leads to morpho-functional microvascular damage that could be improved by some dietary interventions. In this study, the intake of Brazil nuts (Bertholletia excelsa), composed of bioactive substances like selenium, α- e γ- tocopherol, folate and polyunsaturated fatty acids, have been investigated on antioxidant capacity, lipid and metabolic profiles and nutritive skin microcirculation in obese adolescents.
Methods
Obese female adolescents (n = 17), 15.4 ± 2.0 years and BMI of 35.6 ± 3.3 kg/m2, were randomized 1:1 in two groups with the diet supplemented either with Brazil nuts [BNG, n = 08, 15-25 g/day (equivalent to 3 to 5 units/day)] or placebo [PG (lactose), n = 09, one capsule/day] and followed for 16 weeks. Anthropometry, metabolic-lipid profiles, oxidative stress and morphological (capillary diameters) and functional [functional capillary density, red blood cell velocity (RBCV) at baseline and peak (RBCVmax) and time (TRBCVmax) to reach it during post-occlusive reactive hyperemia, after 1 min arterial occlusion] microvascular variables were assessed by nailfold videocapillaroscopy at baseline (T0) and after intervention (T1).
Results
T0 characteristics were similar between groups. At T1, BNG (intra-group variation) had increased selenium levels (p = 0.02), RBCV (p = 0.03) and RBCVmax (p = 0.03) and reduced total (TC) (p = 0.02) and LDL-cholesterol (p = 0.02). Compared to PG, Brazil nuts intake reduced TC (p = 0.003), triglycerides (p = 0.05) and LDL-ox (p = 0.02) and increased RBCV (p = 0.03).
Conclusion
Brazil nuts intake improved the lipid profile and microvascular function in obese adolescents, possibly due to its high level of unsaturated fatty acids and bioactive substances.
Trial Registration
Clinical Trials.gov NCT00937599
doi:10.1186/1743-7075-8-32
PMCID: PMC3123174  PMID: 21619692
microcirculation; obesity; Brazil nuts; adolescents; oxidative stress; lipid profile
19.  Central Nervous System Effects of Iso-6-spectaline Isolated from Senna Spectabilis var. Excelsa (Schrad) in Mice 
The central nervous system (CNS) depressant and anticonvulsant activities of iso-6-spectaline (SPEC) were investigated in animal models. The SPEC from Senna spectabilis var. excelsa (Schrad) (0.1, 0.5 and 1.0 mg/ kg) injected by oral route (p.o.) in mice caused a significant decrease in the motor activity up to 30 days after the administration and in the dose of 1.0 mg/kg significantly reduced the remaining time on the Rota-rod apparatus. Additionally, SPEC (0.1, 0.5 and 1.0 mg/kg, p.o.) was also capable of promoting increase of latency for development of convulsions induced by pentylenetetrazole. This SPEC was also capable of promoting an increase of latency for development of convulsions induced by picrotoxin at highest dose. In the same way, the anticonvulsant effect of SPEC was affected by pretreatment with flumazenil, a selective antagonist of the benzodiazepine site of the GABAA receptor. These results suggest possible CNS depressant and anticonvulsant activities in mice that needs further investigation.
doi:10.4103/0975-1483.83772
PMCID: PMC3159278  PMID: 21897664
Fabaceae; open field; pentylenetetrazole; picrotoxin; senna spectabilis
20.  Antihistaminic activity of aqueous extract of stem bark of Ailanthus excelsa Roxb. 
Pharmacognosy Research  2011;3(3):220-224.
Background:
Biologically active compounds from natural sources are of interest as possible new drugs for different diseases. Over many centuries humans have been mining the bounties of nature for discovering natural products that have been used for the treatment of all human diseases. Ailanthus excelsa Roxb. (Simaroubaceae) is widely used in the Indian system of medicine as an antiasthmatic, antispasmodic, bronchodilator, anticolic pain, anticancer, antidiabetic etc. The plant was also reported for its antiasthmatic, bronchodilatory, antiallegic and many more such activities.
Objective:
To evaluate the antihistaminic activity of aqueous extract of stem bark of Ailanthus excelsa Roxb.
Materials and Methods:
We have studied the effect of aqueous extract of stem barks of A. excelsa Roxb. at a doses 100 μg/mL in the isolated goat tracheal chain preparation in vitro and 100, 200, 400 mg/kg doses orally in passive paw anaphylaxis in rat, clonidine-induced catalepsy in mice models in vivo for its antihistaminic activity.
Results:
Aqueous extract of stem barks of A. excelsa Roxb. significantly (***P<0.001) inhibits the percentage contraction at concentration of 100 μg/mL in goat tracheal chain preparation. A. excelsa Roxb. extract (100, 200, and 400 mg/kg oral) and dexamethasone (0.5 mg/kg, i.p.) also significantly reduced (**P<0.01) the paw volume at fourth hour and the percentage inhibition was found to be 13.98%, 28.49%, 42.47% and 46.77% respectively. The aqueous extract of stem barks of A. excelsa Roxb. (100, 200, 400 mg/kg, p.o.) and chlorpheniramine maleate (10 mg/kg, i.p.) significantly inhibited (*P<0.05, **P< 0.01) clonidine-induced catalepsy in mice at 150 min after the administration of clonidine.
Conclusion:
The aqueous extract of stem bark of A. excelsa Roxb. possess significant antihistaminic activity (H1-antagonist) and can be attributed to bronchodilating, anti-inflammatory, adaptogenic activity etc. Hence detailed study needs to be conducted to evaluate the phytoconstituent responsible for the above mentioned results and their clinical efficacy in the treatment of related diseases.
doi:10.4103/0974-8490.85014
PMCID: PMC3193625  PMID: 22022173
Ailanthus excelsa Roxb; antihistaminic activity; passive paw anaphylaxis
21.  The Genetic Polymorphisms and Colonization Process of Olive Fly Populations in Turkey 
PLoS ONE  2013;8(2):e56067.
The olive fruit fly, Bactrocera oleae, is the most important pest of olives in olive growing regions worldwide, especially in the Mediterranean basin and North America. Despite the economic importance of the olive fly, the colonization route of this species is unclear. We used nuclear microsatellite markers and mitochondrial DNA to provide information about the population structure and invasion route of olive fly populations in Turkey, as representative of the Eastern Mediterranean region. Adult fly samples were collected from 38 sublocations covering all olive growing regions in Turkey. The simple sequence variability data revealed a significant genetic variability in olive fly populations and a certain degree of differentiation between Mediterranean and Aegean populations. Mediterranean populations harbor higher levels of microsatellite variation than Aegean populations, which points to the eastern part of the Mediterranean as the putative source of invasion. mtDNA results suggest olive flies from the western part of Turkey are closely related to Italo-Aegean flies of the Mediterranean basin and the olive fly populations have invaded the northern part of the Mediterranean basin through western Turkey. In addition, finding specific American haplotypes in high frequencies might indicate that Turkey is the possible source of American olive fly populations. In order to more precisely characterize the population structure and invasion routes of this organism, more DNA-based sequence analysis should be carried out worldwide.
doi:10.1371/journal.pone.0056067
PMCID: PMC3573072  PMID: 23457499
22.  Genetic Diversity and Differentiation of Juniperus thurifera in Spain and Morocco as Determined by SSR 
PLoS ONE  2014;9(2):e88996.
Juniperus thurifera L. is an important tree endemic to the western Mediterranean basin that it is able to grow in semi-arid climates. It nowadays exhibits a disjunct distribution pattern, occurring in North Africa, Spain, France and the Italian Alps. The Strait of Gibraltar has acted as an efficient barrier against gene flow between African and European populations, which are considered different subspecies by some authors. We aimed at describing the intraspecific genetic diversity of J. thurifera in populations from the Iberian Peninsula and Morocco and the phylogeographical relationships among these populations. The ploidy level of J. thurifera was examined and eleven nuclear microsatellites (nSSRs) developed for J. thurifera were assessed for genotyping this species. Six nSSRs were polymorphic and subsequently used to assess the genetic diversity and structure of the studied populations. Genotyping of the tetraploid J. thurifera using nuclear microsatellites supports the separation of Moroccan and Spanish populations into two genetically differentiated groups that correspond to the proposed subspecies africana and thurifera. High values of within population genetic diversity were found, that accounted for 90% of the total genetic variance, while population structure was weak. The estimators of genetic diversity were higher in populations of Spain than in populations of Morocco pointing for a possible loss of genetic diversity during the spread of this species to Africa from Europe.
doi:10.1371/journal.pone.0088996
PMCID: PMC3923062  PMID: 24533164
23.  X-chromosome SNP analyses in 11 human Mediterranean populations show a high overall genetic homogeneity except in North-west Africans (Moroccans) 
Background
Due to its history, with a high number of migration events, the Mediterranean basin represents a challenging area for population genetic studies. A large number of genetic studies have been carried out in the Mediterranean area using different markers but no consensus has been reached on the genetic landscape of the Mediterranean populations. In order to further investigate the genetics of the human Mediterranean populations, we typed 894 individuals from 11 Mediterranean populations with 25 single-nucleotide polymorphisms (SNPs) located on the X-chromosome.
Results
A high overall homogeneity was found among the Mediterranean populations except for the population from Morocco, which seemed to differ genetically from the rest of the populations in the Mediterranean area. A very low genetic distance was found between populations in the Middle East and most of the western part of the Mediterranean Sea.
A higher migration rate in females versus males was observed by comparing data from X-chromosome, mt-DNA and Y-chromosome SNPs both in the Mediterranean and a wider geographic area.
Multilocus association was observed among the 25 SNPs on the X-chromosome in the populations from Ibiza and Cosenza.
Conclusion
Our results support both the hypothesis of (1) a reduced impact of the Neolithic Wave and more recent migration movements in NW-Africa, and (2) the importance of the Strait of Gibraltar as a geographic barrier. In contrast, the high genetic homogeneity observed in the Mediterranean area could be interpreted as the result of the Neolithic wave caused by a large demic diffusion and/or more recent migration events. A differentiated contribution of males and females to the genetic landscape of the Mediterranean area was observed with a higher migration rate in females than in males. A certain level of background linkage disequilibrium in populations in Ibiza and Cosenza could be attributed to their demographic background.
doi:10.1186/1471-2148-8-75
PMCID: PMC2315647  PMID: 18312628
24.  Bronchodilator activity of aqueous extract of stem bark of Ailanthus excelsa Roxb 
Pharmacognosy Research  2010;2(2):102-106.
Biologically active compounds from natural sources are of interest as possible new drugs for infectious diseases. Ailanthus excelsa Roxb. has been used in Indian system of medicine in the treatment of asthma, bronchitis, cold, colic pain, etc. Stem bark of A. excelsa Roxb. has been used as a decoction in traditional claims. So, our traditional claims enforced us to evaluate its bronchodilator activity. We have evaluated its bronchodilator activity in milk-induced leukocytosis and eosinophilia, clonidine-induced mast cell degranulation, bronchoalveolar lavage fluid (BALF), and lung histopathology models. The aqueous extract of stem bark in doses of 100, 200, 400 mg/kg showed significant activity.
doi:10.4103/0974-8490.62955
PMCID: PMC3140105  PMID: 21808549
Bronchodilator; Ailanthus excelsa Roxb.; bronchoalveolar lavage fluid
25.  Genetic Diversity and Population Structure of Wild Olives from the North-western Mediterranean Assessed by SSR Markers 
Annals of Botany  2007;100(3):449-458.
Background and Aims
This study examines the pattern of genetic variability and genetic relationships of wild olive (Olea europaea subsp. europaea var. sylvestris) populations in the north-western Mediterranean. Recent bottleneck events are also assessed and an investigation is made of the underlying population structure of the wild olive populations.
Methods
The genetic variation within and between 11 wild olive populations (171 individuals) was analysed with eight microsatellite markers. Conventional and Bayesian-based analyses were applied to infer genetic structure and define the number of gene pools in wild olive populations.
Key Results
Bayesian model-based clustering identified four gene pools, which was in overall concordance with the Factorial Correspondence Analysis and Fitch–Margoliash tree. Two gene pools were predominantly found in southern Spain and Italian islands, respectively, in samples gathered from undisturbed forests of the typical Mediterranean climate. The other two gene pools were mostly detected in the north-eastern regions of Spain and in continental Italy and belong to the transition region between the temperate and Mediterranean climate zones.
Conclusions
On the basis of these results, it can be assumed that the population structure of wild olives from the north-western Mediterranean partially reflects the evolutionary history of these populations, although hybridization between true oleasters and cultivated varieties in areas of close contact between the two forms must be assumed as well. The study indicates a degree of admixture in all the populations, and suggests some caution regarding genetic differentiation at the population level, making it difficult to identify clear-cut genetic boundaries between candidate areas containing either genuinely wild or feral germplasm.
doi:10.1093/aob/mcm132
PMCID: PMC2533604  PMID: 17613587
Olea europaea; genetic variability; gene pools; microsatellites; oleasters; population structure

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