The Limit of detection

(LOD) was defined as the lowest co

The Limit of detection

(LOD) was defined as the lowest concentration to be detected, taking into consideration a signal-to-baseline noise ratio larger than 3 (Marin et al., 2007, Shah et al., 2000 and USDHHS, 2001). According to the FDA Guidance, solvent evaporation stability during storage in the autosampler for a 24 h period was established at five concentrations of 37.50, 25.00, 17.50, 10.00 and 5.00 mg L−1 in triplicate and was tested only for tocopherols. Considering that solvent evaporation would affect the concentrations of tocopherols and carotenes in the same proportion, INCB018424 nmr no specific stability test was required for carotenes. Three Amazon oils were selected: Buriti (Mauritia Flexuosa), Patawa (Oenocarpus bataua) and Tucuma (Astrocaryum aculeatum). Samples were dissolved in hexane and aliquots of 20 μL were injected in the HPLC system. The following fruits pulps were purchased at local markets in the Amazon Region:

Buriti pulp was acquired in Abaetuba (Pará, Brazil), and Patawa and Tucuma pulps in Belém (Pará, Brazil), during harvest time. Thirty fruits of each specie were gathered in three different places which were separated by a distance of at least two kilometers from each other, adding up to 90 fruits from each specie. The Bligh and Dyer (1959) method was used to extract oils from the dried pulps. The total lipid fraction was learn more extracted by exhaustive maceration with

chloroform and methanol, followed by filtration of solids and separation of the solvent/fat layer. Dried samples (10% moisture) were used to facilitate extraction with organic solvents. All data are presented as mean values ±SD and the mean values were analysed by one-way ANOVA and Tukey-HSD triclocarban at p < 0.05 with SAS. Reproducible separation of β-carotene was obtained in the same silica normal-phase column used for tocopherol analysis. Retention time of β-carotene is 1.9 min, showing that this compound has lower affinity with the column. Peaks are sharp, symmetrical and all homologues were efficiently separated (Fig. 1). Tocopherols were analysed using both PDA and fluorescence detectors. Retention times for tocopherols using the fluorescence detector were, respectively, 7.6, 16.6, 19.9 and 29.1 min for the α-, β-, γ- and δ-tocopherol homologues. For the PDA detector, retention times were 7.2, 16.4, 19.3 and 28.5 min, respectively, for the α-, β-, γ- and δ-tocopherol homologues. Note that retention times for PDA were lower than for fluorescence. This difference is due the system configuration: the samples pass through the PDA detector and then the fluorescence detector. It is also important to highlight that retention times can vary slightly on different days and analysis.

moschata ‘Menina Brasileira’ and C maxima ‘Exposição’ pumpkin pu

moschata ‘Menina Brasileira’ and C. maxima ‘Exposição’ pumpkin purees, respectively. For the

C. moschata ‘Menina Brasileira’ samples, the major carotenoids were Stem Cell Compound Library ic50 all-trans-β-carotene and α-carotene, with lower amounts of ζ-carotene, violaxanthin and lutein. In the samples of C. maxima ‘Exposição’, the major carotenoid was all-trans-β-carotene, with good amounts of violaxanthin and lutein in raw pumpkins. Although they are still considered interesting when compared with other plant species, concentrations of carotenoids in raw pumpkins are lower than those reported in other studies regarding the same species and varieties of pumpkins. Azevedo-Meleiro and Rodriguez-Amaya (2007) also noted the all-trans-β-carotene and α-carotene as the major carotenoids in C. moschata ‘Menina Brasileira’ pumpkins, but with higher concentrations, 66.7 ± 9.1 μg/g to all-trans-β-carotene and 26.8 ± 5.1 μg/g to α-carotene. In the C. maxima ‘Exposição’ species, authors noted violaxanthin (20.6 ± 3.3 μg/g)

find more as its major carotenoid. The all-trans-β-carotene was the second in concentration, 15.4 ± 4.2 vs 13.38 ± 2.25 μg/g detected in this present study, where it was the major carotenoid. Indeed, the concentration ranges cited in literature are wide. Rodriguez-Amaya et al. (2008) detected concentrations of 14–79 μg/g of all-trans-β-carotene and 8.3–42 μg/g of α-carotene for C. moschata ‘Menina Brasileira’ pumpkins, and 3.1–28 μg/g of all-trans-β-carotene for C. maxima ‘Exposição’

pumpkins. Major qualitative and quantitative differences in carotenoids, even within the same species and variety, can be noted depending on the cultivar, Rebamipide differences in growing environment, such as temperature, nutrient availability, soil, intensity of sunlight, ripening stage, post harvesting, amongst other factors that can significantly affect the biosynthesis and metabolism of carotenoids in vegetables ( Cazzonelli and Pogson, 2010 and Rodriguez-Amaya, 1999). The studies mentioned above, for example, were conducted with pumpkins harvested in the northeast and southeast regions of Brazil, where the average temperatures are higher than those in the southern region of the country, where the pumpkins used in this study were cultivated. Regarding the effect of processing on the carotenoids, in almost all the cases where a decrease in the concentrations was noted during processing, they occurred mainly in the cooking stage. For instance, for the samples of the C. moschata ‘Menina Brasileira’ pumpkins, besides the disappearance of violaxanthin there was also a decrease of 23.7% in ζ-carotene after cooking. Even after cooking, there was a decrease of 17.9% and of 16.9% in α-carotene and all-trans-β-carotene, respectively, but the concentrations of these carotenes in cooked pumpkins were not considered significantly different (P ⩽ 0.05) from those obtained for raw pumpkins.

Cheeses were triturated and homogenised for the physical chemical

Cheeses were triturated and homogenised for the physical chemical assays. To

characterise cheese samples the following analysis were carried out, in triplicate: fat by the method of Gerber-Van Gulik (Instituto Adolfo Lutz., 1985); titratable acidity (Instituto Adolfo Lutz, 1985); moisture (Case, Bradley, & Williams, 1985); ash (Instituto Adolfo Lutz, 1985); salt (Serres, Amariglio, & Petransxiene, 1973) and to determine pH, 10 g of grated cheese were transferred to a 100 ml beaker, 10 ml of distilled water was added and after homogenising 30 ml of distilled water was added; the mixture was left to rest for 5 min and was then filtered through hydrophilic cotton into 250 ml Erlenmeyer flask, cotton was rinsed with 10 ml of distilled water, squeezed and the clear filtrate was used for pH determination.

As a common practise in our laboratory, analysis for fat, moisture, ash and salt were NVP-BGJ398 datasheet only determined on the first day of ripening. To evaluate proteolysis, total nitrogen (TN) was determined by the micro-Kjeldahl method according to AOAC (1997) using a factor of 6.38 to determine total protein. For soluble nitrogen evaluation, the procedure for preparing the cheese extract was adapted from Vakaleris and Price (1959). Fifty grammes of grated cheese was homogenised with 100 ml of distilled water learn more at 40–45 °C and 50 ml of 0.5 M sodium citrate in a mixer for 7 min. The homogenous milky solution was transferred, using distilled water, into a 250 ml volumetric flask, cooled to room temperature, brought up to volume and thoroughly mixed. This was referred to as the homogenate. To obtain the fraction of nitrogen soluble at pH 4.6, a 100 ml

aliquot of the homogenate was transferred to a 250 ml beaker, 20 ml of 1.41 M HCl was added and after 5 min 15 ml of distilled water was added. The solution was filtered through a Whatman No. 1 filter paper and the clear filtrate was used for subsequent measurement of total nitrogen content. To GABA Receptor obtain the fraction of nitrogen soluble in TCA 12%, a 50 ml aliquot of the homogenate was transferred to a 250 ml beaker, 50 ml of 24% TCA solution was added and after 15 min 15 ml of distilled water was added. The solution was filtered through a Whatman No. 1 filter paper and the clear filtrate was used for subsequent measurement of total nitrogen content. Ripening extension and depth indices were expressed as percentage of total nitrogen: NS-pH 4.6/TN*100 and NS-TCA 12%/TN*100, respectively. Proteolysis was monitored as described by Shalabi and Fox (1987). For this, 20 mg of cheese samples were incubated at 37 °C, in Eppendorf flasks, with 0.4 ml of 0.062 M tris–HCl buffer, pH 6.7, containing 42% (w/v) urea for 1 h. Afterwards, 10 μl of β-mercaptoethanol were added and the mixture again kept at 37 °C for 45 min. Finally, a drop of bromophenol blue was added.

These models synthesize the best understanding of physiological p

These models synthesize the best understanding of physiological processes and vegetation dynamics, to predict terrestrial carbon fluxes, in

response to future global change factors, including eCO2. Collectively, however, such models exhibit a wide range of sensitivities to future conditions (of CO2 and climate) and exhibit asynchronous behavior under different scenarios (Sitch et al., 2008; Galbraith et al., 2010). The outcomes suggest that our present empirical understanding is insufficient, particularly in terms of soil nutrient limitation Selleckchem Kinase Inhibitor Library and ecosystem responses to eCO2 (Fisher et al., 2013). So far, DGVM predictions for eCO2 induced changes in NPP have only been experimentally validated via comparisons with a limited subset of eCO2 experiments in temperate forests Epacadostat solubility dmso (n = 4) ( Sitch et al., 2008 and Norby et al., 2005). Such forests are widely considered to be constrained by soil nitrogen (N) ( Finzi et al., 2006). At a global scale such conditions are atypical, because many regions

are phosphorus-limited ( Lloyd et al., 2001) and also sequester carbon under very different conditions of temperature, precipitation and sunlight availability. The influence of global variations in environmental conditions appears largely untested by eCO2 research, yet historically DGVMs have only been validated on the basis of this limited number of temperate experiments. To improve our confidence in such models, a better understanding is needed to verify how component plant-soil processes respond to and interact with eCO2 at the global scale. Long-term eCO2 experiments in major global regions for C storage and sequestration

are potentially the most direct way of achieving this. We conducted an appraisal of all eCO2 experiments since 1987, using the following combined search terms in an ISI Web of Science search: “elevated CO2,” “FACE,” “CO2 enrichment” and “ecosystem.” Our specific aim was to consider typical experiments relevant to natural ecosystems, so sources were excluded to remove any investigations using controlled environment Levetiracetam chambers or enclosed greenhouses to simulate eCO2 conditions. Similarly, studies were also excluded if their primary focus was on crop species. Our final synthesis identified 675 papers from 151 unique studies (with a 10 m2–3000 m2 range in total experimental plot area) investigating ecosystem-level responses to eCO2 worldwide, since 1987, when the wider adoption of eCO2 methods first emerged for ecological studies. Of these experiments nearly 44% used FACE technology, whereas others utilized open-top chambers (48%), naturally-occurring CO2 springs (5%) or CO2 systems fitted to the branches of entire trees (3%). The FACE system has the least impact on other growing conditions including microclimate, but is inherently costly and may not be suitable in some locations.

However, we added as a second interruption an anagram task Parti

However, we added as a second interruption an anagram task. Participants were shown four or five-letter anagrams along with two letters selleck screening library just below to the left and the right of the anagram (Times font, size = 24). Subjects

had to press either the left or right key to indicate which of these was the first letter of a legal word that could be formed with the anagram letters. Anagrams were selected from a pool of 140 possible words. Words could be used more than once per experiment, but only after all other words had been used. Another difference from the preceding experiments was that interruption task stimuli both for the math and the anagram task were presented in random positions within an area that was at least 6°, but no more than 9° away from the center of the screen. Half of the subjects were randomly assigned to the 1:2 mapping group, which worked either only with the math task or only with the anagram task (i.e., 10 subjects each). The other half of the subjects was assigned to the 2:2 mapping group, for which both the math and the anagram task were presented randomly. Otherwise, the endogenous and exogenous tasks were identical

to the exo/endo conditions in Experiments 1 and 2 with 50% conflict in either the endogenous or the exogenous task. We used the same trial exclusion criteria as in the previous experiments. Again, in no condition of the primary task did error rates exceed 3.0% and in no instance did the pattern

of error effects counteract the pattern of RTs. Therefore, we focus our reporting of analyses on RTs only, but we do present Celecoxib errors in Fig. 5 along with RTs. The mean error rate for the math task was 11.1% (SD = 6.7%) in the group that only performed the math task and 15.5% (SD = 12.4%) in the group that performed both interruption tasks. The corresponding values for the anagram task were 9.8% (SD = 11.4) and 6.1% (SD = 3.4). Mean RTs for the math task were 4486 ms (SD = 1496) for the math-only group and 4979 ms (SD = 2118) for the mixed group. The corresponding RTs for the anagram task were 2759 ms (SD = 789) and 2478 ms (SD = 1028). The upper panel of Fig. 5 presents RT results for the primary tasks as a function of task, interruption, and conflict, separately for the condition in which interruption task and primary tasks were either inconsistently (1:2) or consistently (2:2) mapped. As apparent, across both conditions the qualitative data pattern was largely similar to the one obtained for the corresponding exo/endo conditions in Experiments 1 and 2. The switch-cost asymmetry, that is the Task × Interruption interaction was highly significant, F(1, 38) = 55.88, MSE = 5846.37, p < .001, and this effect was not modulated by the Mapping factor, F(1, 38) = .25. As in the preceding experiments, the cost-asymmetry was further modulated by conflict, F(1, 38) = 26.07, MSE = 5745.93, p < .001.

Forestry has made an important contribution to the Swedish econom

Forestry has made an important contribution to the Swedish economy for many years, which is why the Swedish NFI was started in 1923. The importance of forestry

differs among countries and if it is not a key-category, the IPCC (2003) accepts a higher uncertainty (Tier 1) for reported carbon stock changes. Our evaluation of the consequences of using BEFs relies on the assumption that biomass functions result in good (close to unbiased) results. This assumption rests on the ability of biomass functions to adapt to different conditions (through the measured independent variables) in a manner that BEFs cannot do. Although BEFs are assumed to be constants our results show that they vary substantially over time, and we think that this is an important message to Olaparib mouse people and countries involved in greenhouse gas reporting based on NFI-type data. Although not studied here, the default method might be an alternative approach to the stock change method (IPCC, 2003). When using the default method, changes in biomass for the living biomass pool may be estimated by applying BEFs to growth and drain. We argue that the risk of bias is probably higher when using the default method and will now try to discuss why: The Swedish NFI provides

estimates of stem volume and growth based on bore find more cores extracted from sample trees (on temporary sample plots). To obtain acceptable accuracy, the estimated growth is based on the last five fully developed annual year rings combined

with average data for 5 years. This means that the growth for recent years has to be extrapolated. The drain is probably underestimated as it is difficult in the field to judge whether the harvest occurred within the last year, and a proportion of stumps are usually unidentified; however, we have tried to eliminate Ribonucleotide reductase this underestimation by calibration from stock changes on permanent plots. Alternatively, harvests may be estimated indirectly from consumption or production statistics of harvested wood products. For both growth and drain we expect a large potential bias when converting volume to biomass. This bias may be reduced if separate BEFs are derived for growth and harvest. One advantage of using harvest statistics is the data is reasonably up to date but disadvantages include (i) both legal and illegal export/import need to be considered, (ii) the proportion of pulp that is biomass has to be known, (iii) the data does not account for natural mortality and (iv) harvest cannot be correlated with land use (harvest should be reported and recorded for several KP-activities). Thus, it is likely that the risk of systematic errors is higher using the default rather than the stock change method.

The development of new antiviral molecules derived from acyclovir

The development of new antiviral molecules derived from acyclovir increases the selection pressure risk of resistant strains (Danve-Szatanek et al., 2004) that have been observed in vivo since the first large therapeutic trials ( McLaren et al., 1985). Therefore, the search for new antiviral agents, especially those with different mechanisms of action, is a crucial goal ( Butler, 2008). Alisertib Cardiac glycosides belong to a group of naturally derived compounds that bind to and inhibit Na+K+ATPase (Lingrel et al., 1997). Members of this group have been traditionally used for the treatment of heart

failure and atrial arrhythmia, such as digoxin, digitoxin and ouabain (Rahimtoola and Tak, 1996). Recently, other important applications have been suggested for these compounds related to their potential anticancer (Prassas and Diamandis, 2008) and antiviral activity (Dodson et al., 2007,

Hartley et al., 2006, Hoffmann et al., 2008 and Su et al., 2008). In this report, we screened 65 cardenolide derivatives obtained from plants, by synthesis or by fungi biotransformation, for anti HSV-1 and HSV-2 activity. Among them, glucoevatromonoside (Fig. 1), isolated from a Brazilian cultivar of Digitalis lanata ( Braga et al., 1996) was chosen for its lower IC50 against Wnt tumor HSV to further elucidate its mechanism of action. The 65 tested cardenolide derivatives were obtained from plants (Braga et al., 1996 and Braga et al., 1997), by synthesis (Extrasynthèse, Genay, France; Merck, Darmstadt, Germany; Boehringer, Mannheim, Germany; Carl Roth, Karlsruhe, Germany), Dipeptidyl peptidase or by fungi biotransformation (Pádua

et al., 2005 and Pádua et al., 2007). Acyclovir, digoxin, dextran sulfate and furosemide were obtained from Sigma (St. Louis, MO, USA). All compounds were dissolved in dimethyl sulfoxide (DMSO) (Merck, Darmstadt, Germany), not exceeding the minimum non cytotoxic concentration of 1% DMSO and were further diluted in culture medium prior its use. Vero (ATCC: CCL 81) and GMK-AH1 (Department of Clinical Virology, University of Göteborg, Sweden) cells were grown in Eagle’s minimum essential medium (MEM; Cultilab, Campinas, Brazil) supplemented with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA), 100 U/mL penicillin G, 100 μg/mL streptomycin and 25 μg/mL amphotericin B (Cultilab) and maintained at 37 °C in a humidified 5% CO2. HSV-1 [KOS and 29R (acyclovir-resistant) strains] (Faculty of Pharmacy, University of Rennes, France), and HSV-2 [333 strain (Department of Clinical Virology, Göteborg University, Sweden)] were propagated in Vero and GMK AH1 cells, respectively. Viral stocks were stored at −80°C and titrated based on plaque forming units (PFU) count by plaque assay as previously described (Burleson et al., 1992). Firstly, cytotoxicity was determined by MTT assay (Mosmann, 1983).

As a consequence, E7 quickly leads to the stabilization of p53

As a consequence, E7 quickly leads to the stabilization of p53

and hence the need for E6:E6AP to neutralize p53 or lead to its ubiquitinylation and proteasome-mediated turnover. The selective mechanism of action of CDV as antiproliferative agent could be inferred by analysing the specific signatures identified in CDV-exposed PHKs that were not found in tumor cells, including cell cycle regulation and activation of DNA-double strand breaks (DSBs) repair mechanisms (i.e. ‘ATM Signalling’ and ‘Double-Strand Break Repair by Homologous Recombination’) (Fig. 12B). These findings suggest that CDV can generate double-strand DNA breaks that cannot be repaired Selleck SRT1720 by tumor cells

but well by normal cells (De Schutter et al., 2013c). Furthermore, when we compared the efficiency of CDV incorporation into genomic DNA in the different cell types, higher amounts of CDV were incorporated in the genomic DNA of transformed epithelial cells compared to PHKs, despite the fact that the levels of intracellular CDV metabolites were not significantly different Trichostatin A mw among the cell types investigated. Recently, these findings were confirmed by P. Hadaczek and co-workers who also found that CDV is incorporated into cellular DNA activating DNA damage response pathways due to increased DNA breaks that prompt elevated tumor cell apoptotic response in glioblastoma cells (Hadaczek et al., 2013). Besides differences in cell cycle regulation and DNA repair pathways, our gene expression profiling analysis also allowed the D-malate dehydrogenase identification of other pathways and functions that were induced or repressed following exposure to CDV differently in PHKs compared to HPV-positive and/or HPV-negative cells (De Schutter et al.,

2013c). For instance, Rho GTPase pathways and the acute phase response pathway were solely activated in immortalized cells while normal keratinocytes showed the activation of several metabolic pathways (Fig. 13). Therefore, besides induction of double-strand DNA breaks, CDV showed a differential effect on specific pathways in normal cells compared to transformed cells that may contribute to the activity and selectivity of the drug for tumor cells. Furthermore, in vitro acquisition of resistance to CDV in SiHa cells was found to implicate a variety of cellular functions and pathways linked to cell death, cell growth and differentiation, cellular movement, metabolism, tissue development as well as inflammatory response ( De Schutter et al., 2013b).

77 ± 21 68 (p = 0 01), and it differed significantly from the pla

77 ± 21.68 (p = 0.01), and it differed significantly from the placebo group (p = 0.04). In the KRG group, the OSDI-symptom subtotal improved the most, from 35.42 ± 16.42 to 23.40 ± 18.65 (p < 0.01), which was thought to affect the greater part of the total OSDI score improvement. Compared to the baseline, six of the 12 items were significantly improved in the KRG group after the 8-week supplementation:

three items (painful eye, blurred vision, selleckchem and poor vision) of the OSDI-symptom; two items of OSDI-function (driving at night and working with a computer); and one item (feeling uncomfortable in air-conditioned areas). In addition, five of these items, except blurred vision, displayed significant differences between the KRG and placebo groups. Patients with full-blown glaucoma suffer from the disease itself. However, most patients, particularly those in the early to moderate stages of glaucoma, complain more about their dry eye symptoms caused by topical glaucoma Selleck AZD2281 medication until the disease progressed. Many earlier studies reported that patients with glaucoma suffer a higher prevalence of ocular surface disease than the normal population [7], [8], [9] and [10]. Leung et al [10] found that 59% of patients with primary open-angle glaucoma (OAG) and ocular hypertension (OHT) reported dry eye symptoms, whereas severe symptoms were noted by 27% of these

patients. The authors concluded that a large proportion of the patients with OAG or OHT had signs and/or symptoms of dry eye, and that the presence of dry eye and the use of benzalkonium chloride (BAK)-containing medications may affect quality of life. Our study similarly demonstrated that dry eye is prevalent in patients treated for glaucoma by showing that almost all the participants had OSDI scores consistent with the presence of dry eye symptoms. The cause of DES in patients with glaucoma is thought to be multifactorial and may include an active ingredient and

a preservative, most commonly BAK [9] and [32]. Several previous studies Oxalosuccinic acid reported that BAK may cause inflammation and potentially other ocular diseases, including allergy, blepharitis, DES, and anatomical eyelid abnormalities [33] and [34]. The prolonged use of preserved topical drugs is an extrinsic cause of increased tear evaporation, which induces a toxic response from the ocular surface. BAK has a well-known dose-dependent toxicity and is most commonly used as a preservative in ophthalmic solutions, particularly in antiglaucoma eye drops [33] and [35]. Its cellular toxicity has been demonstrated experimentally in in vitro studies of conjunctiva-derived and corneal cells [36] and [37]. BAK induces the expression of inflammatory cell markers at the ocular surface [38] and causes epithelial cell damage, apoptotic cell death, and a decrease in goblet cell density, resulting in tear film instability and tear hyperosmolarity [39] and [40].

6%, BA) In the BZ the dominant species is P wallichiana (44%, B

6%, BA). In the BZ the dominant species is P. wallichiana (44%, BA), whereas A. spectabilis, Q. semecarpifolia, R. arboreum and Tsuga dumosa together reach 41% of the total basal

area ( Table 5). The Canonical Correspondence Analysis (CCA) for direct gradient analysis (Fig. 5) revealed interactions among tree species composition, human activities and topography. The first axis (eigenvalue = 0.789) expressed an elevation gradient where upper subalpine forest species were clearly separated from the lower subalpine ones. The second axis (eigenvalue = 0.147) expressed a gradient of slope steepness and distance from buildings and lodges (Table 6). Along this gradient, a group of Rhododendron species appeared clearly distinct from the other species. In particular, R. arboreum and Rhododendron campanulatum were present only in less accessible Roxadustat order sites with steep slopes and located far from human

infrastructures. ZD6474 cost The forests of SNP are denser and more diverse than those located in the BZ, where the prolonged and intensive thinning has altered the forest structure and composition. After the institution of the SNP (1979) the increasing demand for firewood was supplied by logging in external areas very close to the park borders (Stevens, 2003). The Pharak region included in the BZ was heavily logged due to a lack of harvesting regulations. The higher mean basal area and tree size in the BZ could be a consequence of felling practices applied by local populations. Carbohydrate Illegal logging, especially of small trees, could be one of the main causes of the lower diversity and density in the Pharak forests. With regard to the influence of environmental variables on forest structure, we found that less dense and poorer stands are located in close proximity to human constructions (mainly tourist lodges). Human impact in this area consists largely of severe forest degradation, due to the overexploitation of small trees from the most accessible

sites. Preferred logging sites, both for timber and fuelwood, are located uphill of the Sherpa villages since wood removal downhill is easier (Stevens, 2003). Similar processes were found in the Sikkim region of India (Chettri et al., 2002), where the best-conserved forests were confined to steeper slopes and far from tourist settlements. The negative relationship of average tree size and species diversity with elevation confirmed that in mountain regions anthropogenic pressure is generally more important at lower altitude and on more accessible sites (Garbarino et al., 2013 and Castagneri et al., 2010). The higher tree species richness found in BZ forests is probably due to their lower elevation, but the environmental trend revealed by the direct gradient analysis is common to both SNP and BZ. Rhododendron species (R. arboreum, R. barbatum, R. campylocarpum, R. campanulatum) are more abundant on less accessible sites with steeper slope and far from human infrastructures.