(1980). The primary difference to the Draize test is that lower volumes of test substances (0.01 ml/0.01 g) (Lambert et al., 1993) are applied to the right-eye of the animal (Maurer et al., 2002), with no forced eyelid closure employed (ICCVAM, 2010b). Test substances are also only applied to the corneal surface and not the conjunctival sac. The test is believed to be less stressful to the tested animal (Jester et al., 2001). Pathological changes are characterized in the cornea, conjunctiva and iris/cilliary body (Maurer et al., 2002). Most LVET data

is based upon surfactant-based mixtures or responses that are associated with mild irritation or non-irritants. This is due to the importance of surfactant use in cosmetic, pharmaceutical and household cleaning products (Davila et al., GABA pathway 1998). However, this website Gettings et al. (1996) investigated LVET in response to severe irritants and

reported an under-prediction of results when compared to Draize data. Since Draize testing is often criticized for its over-prediction of human responses, it is arguable that LVET testing is more accurate (Freeberg et al., 1984, Freeberg et al., 1986a, Ghassemi et al., 1993 and Roggeband et al., 2000). However, LVET is still criticized for its use of animals. In addition, should a negative irritancy result occur using a lower test volume, the standard procedure is to increase the concentration of the drug, effectively resorting back to Draize testing. The Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) recently evaluated the validity of LVET for the replacement of Draize testing. It was

not considered to be a valid replacement nor recommend for prospective ocular safety testing (ICCVAM, 2010b). As a result, LVET has yet to be adopted by any regulatory agency as an alternative test. The reluctance to adopt LVET may be due to the fact that it does not offer the element of “exaggeration” present in Draize testing, that helps to assure public safety (Freeberg et al., 1986b and Ubels and Clousing, 2005). However, retrospective LVET data is still useful to weight-of-evidence approaches. It has been suggested that the “gold standard” for eye irritation should be the human response (Bagley et al., 2006) and that ideally, a testing strategy to determine Resveratrol if a substance is harmful to humans would utilize an extremely high number of human subjects in order to faithfully represent human diversity. They would have to be unknowingly exposed to a substance under realistic conditions and the effects assessed (Hartung, 2009). However, such experimentation is both unrealistic and unethical. As a result, human study data and experiences of potential ocular hazards are only available from either accidental exposure or clinical studies. Unfortunately, accidental exposure data often does not realistically represent the most severe lesions since exposure is often brief due to immediate flushing of the eye.

, 2006). Research was focused on, but not limited to, the pro-apoptotic ( Shankar et al., 2007 and Shankar and Srivastava, 2007a), anti-proliferative ( Bachmeier et al., 2010), anticancer ( Bar-Sela et al., 2010), antiviral ( Rechtman et al., 2010), antiarthritic ( Funk et al., 2006), anti-amyloid ( Ringman et al., 2005), antioxidant ( Glauert et al., 2010), anti-obesity ( Alappat and Awad, 2010) and anti-inflammatory ( Jurenka, 2009) properties of curcumin. The underlying mechanisms of these diverse effects are only poorly understood, however, they seem to involve the regulation of various molecular targets, including

transcription factors (nuclear factor-kB), growth factors (vascular endothelial cell growth factor), inflammatory cytokines (tumor necrosis factor, interleukin 1 and interleukin 6), protein kinases (mammalian target of rapamycin, mitogen-activated protein this website kinases, and Akt) and other enzymes (cyclooxygenase Carfilzomib cost 2 and 5 lipoxygenase) ( Aggarwal and

Sung, 2009 and Zhou et al., 2011). It is important to note that there are substantial controversies regarding the action of curcumin on HIV as well as inflammatory conditions ( Liu et al., 2005 and White and Judkins, 2011). Increasing evidence indicates that cation channels also serve as targets for curcumin, i.e. micromolar concentrations of curcumin inhibit Ca2+-release-activated Ca2+ channel (ICRAC) and K+ channels (Kv and SK4) in human T cells ( Shin et al., 2011), block the Cav3.2 T-type

Ca2+ current in bovine adrenal zona fasciculata (AZF) cells ( Enyeart et al., 2009), bTREK-1 K+ channels ( Enyeart et al., 2008) and the Kv1.4 voltage-gated K+ channel ( Liu et al., 2006) in bovine adrenocortical cells. Curcumin also seems to ameliorate pain Vildagliptin hypersensitivity in rats through a selective blockade of transient receptor potential vanilloid 1 (TRPV1) channels ( Yeon et al., 2010). In contrast to cation channels, which seem to be inhibited by curcumin, chloride channels seem to be activated by the substance. The open probability of the cystic fibrosis transmembrane regulator (CFTR) chloride channel was reported to be increased by curcumin in excised, inside-out membrane patches ( Berger et al., 2005). Accordingly, Wang et al. ( Wang et al., 2005 and Wang et al., 2007) found that curcumin (0.5–10 μm) stimulated ion currents mediated by both wild-type and ΔF508-CFTR in excised membrane patches. These authors pointed out that the structure of curcumin (two aromatic rings separated by a hydrocarbon spacer) is similar to that of 5-nitro-2-(3 phenylpropylamino)benzoic acid-AM (NPPB-AM), which is an uncharged form of the well-known chloride channel blocker NPPB and acts as a CFTR agonist by increasing the channel opening rate. Interestingly, curcumin was also shown to increase the activity of a CFTR mutant (G551D) with an extremely low open probability despite its normal trafficking to the plasma membrane ( Yu et al., 2011).

4 It should be noted that anti-mitochondrial antibody-negative PBC and false-positive anti-transglutaminase antibodies have been reported in this context.19 and 20 As in the case of AIH, the impact of gluten avoidance is not well established, but it is determinant to improve symptomatic CD and to prevent complications.2 and 20 A relation between CD and PSC has been suggested in several case reports and in a population-based study. However the

strength of this association GDC-0449 purchase is not clearly determined and the benefit of gluten exclusion from the diet was not yet demonstrated.2 and 13 Nonalcoholic fatty liver disease (NAFLD) and steatohepatitis (NASH) are common disorders in the general population and in celiac patients. I-BET-762 research buy Studies found a prevalence of CD in about 3% of individuals with

NAFL and NASH.21 Obesity, a major risk factor for nonalcoholic liver disease, is common in patients with CD not only after but also before gluten withdrawal, which could explain the association between these disorders.22 Additional etiopathogenic mechanisms may be the increased intestinal permeability, resulting in bacterial translocation and production of proinflammatory factors, and malabsorption leading to chronic deficiency of lipotropic molecules.23 and 24 The correlations among CD, obesity and liver disease must be taken into account when establishing the diagnosis and treating celiac patients presenting with elevated liver enzymes. Acute liver failure and advanced liver disease deserve a special consideration. There are several cases reported in literature and CD was found to be from up to 10 times more frequent among patients with chronic liver disease than in the general population.25 The study by Kaukinen and colleagues12 found a high prevalence of CD (4.3%) in patients who underwent liver transplantation. Autoimmune disorders, such as PBC, PSC and AIH were the main etiologies of end-stage liver disease leading to transplantation. This study also describes 4 cases of patients with advanced liver disease

who were found to have CD, all of them improving significantly their liver function with gluten withdrawal. Some of the patients in both groups had no apparent symptoms or signs suggesting CD. The authors emphasize that the early detection and treatment of CD may prevent the progression to end-stage liver failure. Therefore, CD must be screened in patients with autoimmune liver disease or hepatitis/cirrhosis of unknown etiology and in those undergoing liver transplantation. Moreover, an essential component of the clinical surveillance after transplantation in CD patients is the assessment of compliance with a gluten-free diet. The present case illustrates the association between CD and liver disease. Our patient was a young woman presenting with asymptomatic hypertransaminasemia. The initial CD screening was based on autoantibodies, followed by duodenal biopsy.

2B, right panel). DOPE conjugation to LC3 is shown as comparison (Fig. 3B). The results demonstrate that oxidized PE is an effective substrate for LC3 lipidation, although in this case, it is equally effective as the unoxidized parent lipid. To examine for changes in cellular lipid profiles resulting from 12/15-LOX deficiency, UMI-77 molecular weight lipidomics profiling of all phospholipid classes and cholesteryl esters was undertaken on lipid extracts from macrophages

obtained from naïve wild type and 12/15-LOX−/− macrophages. There was a tendency overall for increased PE, PI and cholesteryl esters, but decreased PC in 12/15-LOX deficiency. On the other hand, PA, PS and PG were not different. This suggests that loss of the enzyme results Dabrafenib manufacturer in a selective defect in particular phospholipid classes at the expense of others (Fig. 3). Herein, we show that deficiency of the lipid-oxidizing enzyme, 12/15-LOX, is associated with altered cellular membrane structure. We also demonstrate that

a LOX-derived oxidized phospholipid is an effective substrate for lipidation of both LC3 and Atg8, being preferred over the unoxidized analog in the case of the yeast homolog. This is suggestive of this pathway being involved in regulation of membrane dynamics. Last, we show altered phospholipid content in murine macrophages deficient in 12/15-LOX. Our observations of double membrane structures suggestive of autophagosomes propose a role in autophagy. Normal LC3 expression and lipidation indicate that the defect in the 12/15-LOX−/− macrophages is likely to be upstream of LC3 activity

itself. 12/15-LOX was first described as the human homolog, 15-LOX1, as being highly induced in bleeding anemia in rabbits, inducing significant peroxidation of intracellular membranes that coincided with disappearance of organelles [19], [20], [21], [22] and [23]. Thus, it was proposed as being critically required for reticulocyte maturation into erythrocytes. However subsequent to this, mice deficient in the functional homolog, 12/15-LOX were shown to have normal red cell counts, and interest in this pathway waned [24]. This does not exclude see more that the knockout mice have developed a compensatory mechanism, and that the enzyme still plays a role in normal turnover of organelles during homeostasis. In support of a role for LOX in processes that involve membrane remodeling, previous studies have shown that 12/15-LOX−/− macrophages are unable to undergo a full phagocytosis response towards apoptotic thymocytes [25]. The multiple differences between wild type and 12/15-LOX−/− macrophages seen, including abnormal mitochondria, multiple lysosomal storage bodies and suspected autophagosomes are consistent with LSDs [26], [27], [28] and [29]. Lysosomes are small vesicular organelles, their primary function being to merge with late endosomes to digest their content [30], [31] and [32]. Endosomal degradation is carried out by numerous lipid and protein hydrolases.

2). The results of liver tests were: total bilirubin 195 μmol/L (NR: <22) with 124 μmol/L of conjugated, alanine aminotransferase (ALT) 1833 IU/L (NR: 10–66), aspartate aminotransferase 1467 IU/L (NR: 15–46), alkaline phosphatase (ALP) 86 IU/L (NR 38–136), gamma-glutamyl transferase 68 IU/L (NR: 12–58) and LDH 1531 IU/L (NR: 313–618). Electrolytes, serum albumin, iron and transferrin saturation were normal. IgM anti-HAV, HBsAg, IgM anti-HBc and anti-HCV antibodies were negative. Anti-HIV, anti-CMV, anti-EBV and anti-HSV were also negative. Auto-antibodies (ANA, ANCA, Anti-LKM, AMA and ASMA) were negative.

24 h-urinary copper, ceruloplasmin, α-fetoprotein and α-1 antitrypsin were within normal range. Liver ultrasonography showed no significant abnormality except for increased echogenicity. A liver biopsy by percutaneous

route was then performed without complications. The biopsy material was fragmented and had lesions located in the portal spaces see more and hepatic lobules. The histological examination showed expansion of the portal spaces with scant fibrosis and intense inflammatory infiltrate composed by lymphocytes, eosinophils and few neutrophils. There were also focal lesions of interface hepatitis (Fig. 1). The hepatic lobules showed moderate inflammatory infiltrate, similar to the one noticed in the portal spaces, ballooning degeneration of the hepatocytes (Fig. 2) and isolated apoptotic bodies throughout the entire studied material. It was observed focal hepatic necrosis with collapse TGF-beta inhibitor of the parenchyma, more severe in the perivenular zone, along with discrete fibrosis. There was also focal hepatic steatosis. No granulomas were detected. These findings were consistent with acute hepatitis and were highly compatible with toxic/pharmacological etiology. A gradual decrease in liver enzymes was seen; total bilirubin

continued to rise and reached a peak 40 days after the intake of fosfomycin, and then it also started to decline (Fig. 3). The patient improved symptomatically, in parallel with the decline in aminotransaminases. Three months after fosfomycin intake, all liver function tests had normalized (Fig. 3). Two years after, the patient remains asymptomatic and without alteration of the liver enzymes. Amisulpride Fosfomycin is a widely used, broad-spectrum antibiotic, which exhibits a rapid bactericidal activity against a large number of aerobic Gram positive and Gram-negative bacteria.1 and 2 It is approved as a single-dose therapy (3-g oral dose) for the treatment of uncomplicated urinary tract infections (acute cystitis) in women.7 Usually, it is a well-tolerated drug and does not appear to cause serious reactions. Reported adverse events are usually mild and last only 1–2 days, resolving without treatment. The overall incidence of side-effects is 6% with oral therapeutic dosing and 17% with parenteral administration. Gastrointestinal complaints, mostly diarrhea, are the most frequent. Dizziness, headache and vaginitis have also been reported.

Gallic acid, protocatechuic acid and ellagic acid had UV–vis spectra analogous to hydroxybenzoic acids, due to the presence of benzoyl groups that formed a chromophore with absorption spectra ranging from 255 to 280 nm (Abad-García, Berrueta, Garmón-Lobato, Gallo, & Vicente, 2009). The flavonols quercetin www.selleckchem.com/products/Gefitinib.html and kaempferol gave an intense band I at 347–370 nm and band II at 250–267 nm, due to the substitution of hydroxyl group at carbon 3 of the C ring (Abad-García et al., 2009). Rutin, which is a glycoside of quercetin, gave the same intense

bands I or II as its aglycone (quercetin) (Abad-García et al., 2009). The LOD and LOQ of each polyphenolic compounds were calculated and tabulated in Table 1. Quantification of the polyphenols in the leaves and stems of B. racemosa is presented in Table 2. Overall, the leaves have higher amounts of polyphenolic compounds than the stems. In addition, the amounts of bound phenolics were approximately 20% more than the free phenolics. The polyphenols in the leaves of B. racemosa in

descending order were gallic acid > ellagic acid > quercetin > protocatechuic acid > rutin > kaempferol. In contrast, only three polyphenols were detected in the stems, in the order of gallic acid > ellagic acid > protocatechuic acid. A previous study reported the leaves of B. racemosa, extracted with acidified methanol, to contain 172 μg/g dw of gallic acid, 59.1 μg/g dw of rutin and 5.75 μg/g dw of kaempferol which were lower than Galunisertib concentration our values ( Hussin et al., 2009). In addition to the extraction method, the differences in polyphenolic content may have also been due to variation in pedoclimatic and agronomic conditions ( Manach, Scalbert, Morand, Rémésy, & Jiménez, 2004). In plants, phenolic acids are usually coupled with the cell wall complexes or form ester and glycosidic linkages with organic compounds, such as glucose, quinic, maleic and tartaric acid and terpenes Metalloexopeptidase (Chew, Khoo, Amin, Azrina, & Lau, 2011). Flavonoids can occur in plants

as both aglycones and glycosides, with the latter in higher amounts (Sakakibara, Honda, Nakagawa, Ashida, & Kanazawa, 2003). Acid hydrolysis functions to degrade the ester and glycosidic bonds of polyphenolic compounds, providing a rapid estimation of the amounts of free and bound polyphenols in plant samples. Tannin is an important chemical constituent in B. racemosa ( Bandaranayake, 2002). The hydrolysable tannins are complexes of hydroxybenzoic acids, which can be classified into gallotannins and ellagitannins, derived from the glucose esters of gallic acid and ellagic acid, respectively ( Ignat et al., 2011). Our results showed that there was more bound gallic acid and ellagic acid, compared to the aglycone forms, indicating that most of these acids are in the form of hydrolysable tannins. Quercetin and kaempferol only existed in the plant in their conjugated forms and not as aglycone ( Table 2).

The Forskolin cost SCFA concentrations were determined using a GC 2010 gas chromatograph (Shimadzu Scientific Instruments Inc., Kyoto, Japan) equipped with a flame ionisation detector (FID). One gram of caecal content was thawed and suspended in 5 ml H2O and homogenised for about 3 min. After that, the pH was adjusted to 2–3 by adding 5 M HCl and the solution was kept at room temperature for 10 min with occasional shaking. The suspension was centrifuged (20 min; 3000 rpm) and 1 μl of the supernatant was injected onto a

Nukol-fused silica capillary column (30 m × 0.25-mm i.d., 0.25-μm film thickness; Supelco, Bellefonte, Palo Alto, CA, USA). The column temperature was held at 100 °C for 0.5 min, increased from 20 °C/min to 200 °C and finally held for 5 min. Hydrogen was used as the carrier gas at a flow rate of 1.8 ml/min, and the split ratio was 1:2. Injector and FID temperatures were 200 °C and 240 °C, respectively. Individual fatty acids were identified by comparison with the retention times of standards (Volatile Free Acid Mix, code. 46975; Sigma Chemical Co., St. Louis, MO, USA) and quantified using GC Real Time Analysis 1 software (GC

Solution version 2.30.00, LabSolutions, Shimadzu Scientific Instruments Inc., Kyoto, Japan). The data analysis was carried out with SPSS (SPSS Inc., Chicago, Illinois, USA) for Windows (version 11.5, 2002). All tests were learn more performed assuming bilateral hypotheses and a 5% significance level. Initially, descriptive statistics were used to evaluate the mean and standard deviation (SD) of the studied variable. Data are shown as mean ± SD. In the depletion period, comparison of mean values between CON and ID groups was performed by using an unpaired t-test. In the repletion period, the variable means of the groups were compared by using analysis of variance (ANOVA). A Tukey’s post hoc test was applied to identify where significant differences occurred. A non-parametric Kolmogorov–Smirnov test was applied to verify

the normality of the observations and, when the normality hypothesis Glutathione peroxidase was rejected, an unpaired t-test and ANOVA were substituted with non-parametric Mann–Whitney and Kruskal–Wallis tests, respectively. The observed power was 85–95% for most tests. A discrete to moderate microcytosis and marked hypochromia was observed in the ID rats when compared to those in the CON group (mean corpuscular volume of 64 ± 9.7 and 40.3 ± 6.3 fL; mean corpuscular Hb of 19.2 ± 3.2 and 11.8 ± 0.7 pg for CON and ID groups, respectively; P = 0.001) with significant reductions in Hb concentration and in the Hb Fe pool (P < 0.001; Table 2). These changes were the result of a marked reduction in the serum Fe levels and in transferrin saturation (P < 0.001) as well as in liver Fe stores (P < 0.001).

3 μg/m3 [18.6–22.0] to 33.7 μg/m3 [32.2–35.2] with pooled value of 27.0 μg/m3 [21.7–32.2] (I2 = 81%). The pooled mean estimates of the short-term limit values in the five sensitivity analyses showed little variation. Difference from main analysis ranged from − 1.2 to 1.3 μg/m3 for

PM10; − 2.2 to 1.7 μg/m3 for PM2.5; − 0.4 to 3.6 μg/m3 for NO2; 0 to 0.1 μg/m3 for SO2; − 3.2 to Lenvatinib purchase 3.9 μg/m3 for O3 (Table 2a and Table 2b). When individual cities that contributed significantly to the overall heterogeneity were excluded, the pooled values for PM10 (47.7 μg/m3) and PM2.5 (26.4 μg/m3) were even closer to the WHO-recommended STAQG of 50 and 20 μg/m3 respectively but such changes were negligible for NO2. Our results demonstrate that there is a robust deterministic relationship in the current WHO short-term AQG for PM10 (50 μg/m3) and PM2.5 (25 μg/m3) and their annual guideline targets of 20 μg/m3 and 10 μg/m3 respectively. However, on the basis of this analysis, the short-term AQG of 200 μg/m3 for NO2 cannot provide a regulatory guideline consistent with the annual AQG of 40 μg/m3. This is a pilot study which has formally examined the validity of the short-term limits as predictors of average annual ambient levels of pollutants. The quantified relationships derived from the assumption of a log probability

this website density function for PM10 and PM2.5 indicate good agreement with WHO expert judgment based on a systematic review of scientific

evidence. The physical explanation for lognormality as an appropriate distribution for air pollutant (Ott, 1990) supports our function of geometric mean and standard deviation. The apparent discordance between the WHO short-term and annual AQG for NO2 warrants further study to support revision of the guidelines. Based on evidence of adverse health effects of exposure to low levels of NO2 in adults and infants, WHO has been aware of the need to lower the current annual AQG below 40 μg/m3 for NO2 (WHO, 2006d). If the setting of the annual AQG was correctly specified in terms of reduction of avoidable morbidity, Farnesyltransferase then the required short-term AQG would predictably be even lower than our pooled estimate of 141 μg/m3. However, if the current WHO short-term AQG of 200 μg/m3 for NO2 is complied with in environments represented by the cities in our sample, then the annual mean would be predictably higher than the currently recommended limit of 40 μg/m3, which has already been considered to be insufficient for child health protection (WHO, 2006d). As epidemiological studies have identified different adverse health outcomes from both short- and long-term exposure to air pollution, it is important to maintain the two limits to support a public health evidence-based approach, while remaining open to new hypotheses and the need for revision.

, 2004). These different patterns of resource use efficiency (ReUE) might be explained by the ability of a tree to acquire 3-deazaneplanocin A chemical structure resources. As long as enough resources are available (i.e. canopy closure is not reached) all trees of a stand are equally efficient (Binkley et al., 2002, Binkley, 2004 and Fernández and Gyenge, 2009). When inter-tree competition starts,

larger trees are able to acquire enough resources, whereas smaller trees might already reach their resource compensation point (minimum resource quantity needed to produce a positive growth). That implies an increase in ReUE for larger trees but a decrease in ReUE for smaller trees-supporting the pattern in this study. For Ponderosa pine (Pinus ponderosa C. Lawson), Fernández and Gyenge (2009) observed differences in the water use efficiency before canopy closure, indicating that differentiation in efficiency is defined in earlier

stages (before canopy closure) to ABT-199 datasheet determine the dominant and suppressed trees. A comparison between the thinned and the unthinned treatments revealed that (i) on a tree-level basis, with a given tree size, trees from the unthinned plots were more efficient (except the mature stands) and (ii) on the stand-level, the mean tree of the thinned stand was either more efficient (mature and pole-stage1), as efficient (pole-stage2), or less efficient (immature) than the mean tree of the unthinned plots. Wang (1988) found that dry matter per APAR was not affected by thinning, but rather

from nitrogen fertilization for plots of Sitka spruce. Forrester et al. (in press) showed that for Eucalyptus nitens plantations, LUE in terms of annual above ground biomass increased with thinning, while LUE in terms of wood mass declined. They speculate that a decline of efficiency with thinning may occur on sites that are limited by resources other than light. When analyzing light regimes, we had science to assume that water and nutrient supply was ample, which may not have been the case for the immature stand (the only plot showing a decrease of efficiency with thinning). Assuming that the trees are a representative sample for one hectare, one could roughly scale up to a hectare-level by multiplying the mean efficiency with the stems per hectare. This gives a clear pattern, proving that due to the higher stem number per hectare, the unthinned treatment is always more efficient (with 12.2%, 80.3%, 152%, 185% for mature, immature, pole-stage1 and pole-stage2, respectively). That would mean that more wood per unit light is produced in an unthinned stand. However, forestry typically focuses on producing high quality saw-log timber that cannot be obtained without thinning. Hence a trade-off has to be found between growing the most efficient trees at a low risk of damage with the amount of trees per unit area.

, 2007). The seed production of many agroforestry trees is often informal and very few countries have included these species in their tree improvement programmes. Germplasm of exotic tree species, typically from introductions of unknown provenance and uncharacterised performance, is often collected by smallholders directly for their BMS-387032 mw own planting.

Lillesø et al. (2011), for example, identified five sources for farmers’ tree planting material (farmland, natural forest, plantations, seed orchards and vegetative propagules) and indicated heavy reliance on the first source, with natural forest sources being underutilised. Farmers and local seed dealers often prefer to collect seed from previously Cobimetinib mouse introduced exotic trees in farmland rather than source externally because the transaction costs are lower, even when better-performing seed sources of the same trees may be available elsewhere (Lengkeek et al., 2005 and Muriuki, 2005). In recent decades, there has been a greater focus on the cultivation of indigenous tree species in agroforestry systems, with the involvement of local people in carrying out genetic selection for tree characteristics of importance

to them. One such approach, known as participatory domestication, has been developed in Africa on indigenous fruit trees (see Dawson et al., 2014, this special issue). The advantage of this approach is that genetic quality as a concept is explicitly considered, and local wild stands provide significant genetic variation that is a pool for selection

(Tchoundjeu et al., 2006). The risk of spreading pests and diseases while transferring reproductive material is often considerable. Pests and diseases travel in different substrates and it is challenging Vorinostat concentration to monitor the way they spread; for example, to reconstruct the exact pathways of their past movements. In Europe, Santini et al. (2013) reconstructed the most probable pathways of alien invasive forest pathogen spread since 1800. They found that living plants (57% of all pathogen introductions) and wood (10%) were likely major vectors for introductions, while the share of any other pathway, such as bark, seed, soil and cuttings, was less than 10% over the last two centuries. According to the same authors, over the last few decades, the invasion rate of alien forest pathogens has increased exponentially in Europe, with soil recently becoming a major transfer substrate second to living plants. In the USA, a similar study attributed 69% of the introductions of non-native forest insects and pathogens since 1860 to the trade in living plants (Liebhold et al., 2012). These studies confirm the need for phytosanitary regulations and their careful implementation while transferring tree germplasm. However, they also show that the pathogen risk associated with transferring seed is considerably lower than the risk connected with transferring other materials such as living plants or wood.