Cross sections surveyed by Mendocino County Water Agency between 1996 and 2005

further downstream at Mountain View Bridge indicate fluctuations typical of short-term geomorphic change, with ∼0.8 m of incision during the water year 1998 flood, followed by an increase in bed elevation back almost to the 1996 level in 2001. Between 2001 and 2013, incision lowered the bed by about 0.37 m. Bed elevation fluctuation of erosion or deposition during any one flood is common and longer-term monitoring data is warranted to assess trends. Measurements in a reach of Robinson Creek ∼2.4 km upstream of the mouth measured incision using exposed GSK1120212 order roots of riparian California Bay Laurel Trees as an indicator. In this location, the root systems of numerous trees are fully exposed along both banks of the incised channel. Measured bank heights between the channel bed and the surface of the lateral roots in 2008 reached 2.0 m on average (Fig. 6A). Because trees establish on level alluvial surfaces such as on a creek’s floodplain, vertical banks present below the tree’s root systems clearly indicate incision. In 2013, we assessed tree rings in a core from one of

the undercut trees (main stem diameter ∼198 cm) and assume it is representative of numerous nearby undercut trees of similar size. Portions of the core are indistinct, Alectinib concentration including the heart of the core (Fig. 6B); and because the tree rings are not cross correlated or dated, the core does not give an absolute age. However, about 83 rings are visible, suggesting that the tree established prior to 1930. Because these trees can reach 200 years when mature, we estimate these stream-side trees established sometime after about 1813 and before 1930—and that incision began after their establishment. We examined incision in the study reach through surveyed thalweg, bar crest, and top of bank/edge terrace elevation profiles (Fig. Tacrolimus (FK506) 7A). The thalweg profile has a reach average slope of ∼0.012. Contrasting the

three channel segments between bridges (Table 1) illustrates that the downstream portion of the reach is steeper than the upstream portion. Variation in bed topography is present despite incision; the thalweg profile exhibits irregularly spaced riffles and pools (Fig. 7A). However, pools present have relatively shallow residual depths (the maximum depth of the pool formed upstream of a riffle crest; sensu Lisle and Hilton, 1999); 60% have a residual depth less than 0.6 m. Several pools contained water during the summer of 2005 and 2008 when the majority of the channel was dry. Variation in bed topography is also exhibited in steeper than average apparent knickzones, with slopes of ∼0.018 ( Fig. 7A). Bars are present in the channel (Fig. 7A); the reach averaged bar crest slope is similar to the thalweg slope, 0.012. Average bar height is ∼0.6 m above the thalweg.

Our results confirm that, by

exporting contaminated particles originating from the main inland radioactive plume, coastal rivers are likely to have become a significant Dasatinib supplier and perennial source of radionuclide contaminants to the Pacific Ocean off Fukushima Prefecture. This could at least partly explain the still elevated radionuclide levels measured in fish off Fukushima Prefecture (Buesseler, 2012). Quantification of the hydro-sedimentary connectivity between hillslopes and the identified sinks in the three coastal catchments provided additional information on the timing of sediment transfer processes and their preferential pathways observed along the investigated rivers (Fig. 6). Paddy fields located in the upstream part of both Nitta

and Mano River catchments were well connected to the thalweg and they constituted therefore an important supply of contaminated material to the rivers or to small depressions located in the floodplain. In contrast, in the flat coastal plains of those catchments, large cultivated surfaces were poorly connected to the rivers. A distinct situation was observed in the Ota River catchment. In the upper part of this catchment, land use is dominated by forests that are much less erodible than cropland, but that could deliver contaminated material to the river during heavy rainfall (Fukuyama et al., 2010). Furthermore, the high slope gradients observed in this area may have led to the more frequent occurrence of mass movements in this area. This contaminated material was then stored in the large Yokokawa reservoir (Fig. 6a). In the downstream part of the Ota River catchment, paddy Raf inhibitor fields located in the vicinity of rivers were well buy Staurosporine connected to the watercourses which contrasts with the situation outlined in the coastal

plains of the Mano and Nitta River catchments (Fig. 6b). This transfer timing and preferential pathways are confirmed when we plot the contamination in total 134+137Cs measured in sediment collected during the three fieldwork campaigns along the longitudinal profiles of the investigated rivers (Fig. 7). Overall, we observed a general decrease in the contamination levels measured between the first and the last campaign, especially in the Nitta River catchment (Fig. 7, left panels) where the difference is particularly spectacular along the upstream sections of the Nitta (Fig. 7; profile c–d) and Iitoi Rivers (Fig. 7; profile g–e). Our successive measurements suggest that there has been a progressive flush of contaminated sediment towards the Pacific Ocean. However, the mountain range piedmont and the coastal plains that have remained continuously inhabited constitute a potentially large buffer area that may store temporarily large quantities of radioactive contaminants from upstream areas. However, our data and the drawing of the longitudinal profiles suggest that this storage was of short duration in the river channels.

9A). Consistent with this, Rb2 and Rd significantly reversed EtOH-mediated Sirt1 and PPARα suppression (Fig. 9B). The results suggest that RGE and its major ginsenosides inhibit alcohol-induced fatty liver and liver injury through the recovery of homeostatic lipid metabolism in the liver. ALD, which ranges from simple fatty liver to cirrhosis and hepatocellular carcinoma, remains a major cause of liver-associated mortality worldwide [29]. Early research on the pathogenesis of the

ALD primarily focused on alcohol metabolism-related oxidative stress, malnutrition, and activation of Kupffer cells by endotoxins [30] and [31]. Recently, the characterization of intra- and intercellular signaling pathways, innate and adaptive immune responses, epigenetic features, microRNAs, and stem cells has improved our knowledge of the pathobiology of ALD [31]. LY2109761 concentration Despite improved understanding of the pathophysiology of ALD, there is no Food and Drug Administration-approved drug for the specific treatment of ALD. Therefore, the development of effective therapeutic strategies for ALD is Selleckchem Regorafenib pivotal. KRG has been shown to exhibit several beneficial effects in the treatment of liver diseases through the regulation of immune function and antioxidant activity [16]. However, the effects of KRG on alcohol-induced hepatic steatosis and oxidative stress have not been fully established. Here, we established

the effects of RGE on alcohol-induced liver injury in vivo and in vitro and identified the major component of KRG with beneficial effects in ALD. Ginseng saponins, referred to as ginsenosides, play a major

role in most pharmacological actions of ginseng; however, until now, the role of ginsenosides on EtOH-induced fat accumulation has remained observed. Interestingly, the ginsenosides Rb2 and Rd, but not Rb1, significantly restored EtOH-induced Sirt1 and Erastin PPARα suppression ( Fig. 9B), consistent with RGE treatment to the mice. Moreover, the ginsenosides Rb2 and Rd inhibited EtOH-induced fat accumulation in AML12 cells ( Fig. 9A). The increased lipolytic gene expression and inhibition of fat accumulation resulting from treating by RGE and its major ginsenosides indicates that RGE may be a promising hepatoprotective candidate against liver injury. During the last 5 decades, several animal models of ALD have been studied, which has helped us understand the molecular basis of ALD. The most widely used model for ALD is the Lieber–DeCarli EtOH-containing diet, which is a liquid diet-based voluntary feeding model. Recently, we have developed and reported a more severe alcohol-induced liver injury model (a chronic–binge EtOH model in mice), which is similar to drinking patterns in ALD patients who have a background of long-term drinking (chronic) and a history of recent heavy alcohol use (binge) [25] and [26].

Moreover, many villagers are abandoning swidden rice cultivation Palbociclib price because of increasing land constraints, lower yields, loss of soil fertility and lack of labour availability (Sowerwine, 2004a). Since 1991, much of this land has been declared “watershed protection land”, and swidden rice varieties are rapidly abandoned as more time is devoted to wet rice production (Sowerwine, 2004a). Because of diversification in alternative economic activities, rural households are becoming less dependent on natural resources for their survival,

and deforestation was reduced. This decrease in land pressure after tourism development is not confirmed by previous studies in Southeast Asia, where the presence of alternative income sources has increased the ALK inhibition frequency of cultivation through hired rural labour and/or the expansion of the cultivated area through land purchase (e.g., Forsyth (1995) for northern Thailand). This suggests that local and national land use policy likely plays an important role in directing

tourism development towards sustainable natural resource management. In Sa Pa, conservation policy has had a positive effect on forest protection as most of the forests within the National park remained intact during last the 21 years. This makes the area attractive for tourists , and tourists are further supporting biodiversity conservation by providing extra revenue for conservation. Direct revenue is presently being raised by the Ham Rong project, and by the charging of fees for climbing Fansipan mountain or visiting exclusive sites within Sa Pa district (Frontier Vietnam, 1999). This paper aimed at better understanding of the human–environment interaction in the Sa Pa district after the advent and growth of the tourism industry. A land cover change analysis between 1993 and 2014 showed that the

Sa Pa district as a whole experienced a forest transition, with an observed turning point around mid 2000s. However, trends at district level mask substantial heterogeneity at village level. The results from this paper show that forest cover changes are different in rural villages that have access to alternative others income sources, either from cardamom cultivation under forest canopy or from tourism activities. These rural villages are typically characterized by higher rates of land abandonment and lower rates of deforestation. Because of diversification in alternative economic activities, rural households are becoming less dependent on natural resources and agricultural products for their survival. Our results suggest that the creation of off-farm jobs in the tourism sector, construction or manufacturing can be a driver of shifts in coupled human–environmental changes.

Moving to the south, we encounter the palaeochannels CL1 and CL2, described in the last section. Between the Vittorio Emanuele III Channel and the Contorta S. Angelo Channel there are a few palaeochannels meandering mainly in the west–east direction. These palaeochannels probably belong to another Holocene path of the Brenta river close to Fusina (depicted in Fig. 4. 68, p. 321, in Bondesan and Meneghel, 2004). In

the lower right hand side of the Selleckchem Palbociclib map, we can see the pattern of a large tidal meander that existed already in 2300 BC that is still present today under the name Fasiol Channel. Comparison with the 1691 map shows that the palaeochannels close to the S. Secondo Channel disappeared, and so did the palaeochannel CL1 (Fig. 4b). The palaeochannel CL2 is no longer present in our reconstruction, but it may still exist under the Tronchetto Island, as we observed in the last section. The acoustic areal reconstruction of CL3 overlaps well with the path of the “coa de Botenigo” from the 1691 map that was flowing into the Giudecca Channel. This channel is clearly visible also

in Fig. 4c and click here d. On the other hand, the palaeochannels close to the Fusina Channel of Fig. 4a have now disappeared. This may be related to the fact that in 1438 the Fusina mouth of the Brenta river was closed (p. 320 of Bondesan and Meneghel, 2004). To the lower right, the large meander of the Fasiol Channel is still present and one can see its ancient position and continuation. In 1811, the most relevant changes are the disappearance of the “Canal Novo de Botenigo” and of the “Canal de Burchi” (in Fig. 4c), that were immediately to the north and to the south of the Coa de Botenigo in Fig. 4b, respectively. The map in Fig. 4d has more details with small creeks developing perpendicular to the main channel. Moreover, the edification of the S. Marta area has started, so the last part of the “Coa de Botenigo”

was Y-27632 chemical structure rectified. Finally, the meander close to the Fasiol Channel is now directly connected to the Contorta S. Angelo Channel. In the current configuration of the channels, the morphological complexity is considerably reduced (Fig. 4e). The meanders of the palaochannel CL3 (“Coa de Botenigo”) and their ramification completely disappeared as a consequence of the dredging of the Vittorio Emanuele III Channel. The rectification of the palaochannel CL3 resulted in its rapid filling (Fig. 2d). This filling was a consequence of the higher energetic regime caused by the dredging of the new deep navigation channels in the area. The old Fusina Channel was partially filled and so it was the southern part of the Fasiol Channel meander. The creeks developing perpendicular to the main palaeochannels in 1901 (Fig. 4d) completely disappeared. A more detailed reconstruction of the different 20th century anthropogenic changes in the area can be found in Bondesan et al.

[12]. Peptide model of Sc-ALF created using SWISS-MODEL server consisted of two α-helices crowded against a four-strand β-sheet. Two of the β-strands are in turn linked by a disulfide bond to form an amphipathic loop rich in cationic amino acid side chains ( Fig. 1B). Multiple alignment performed for Sc-ALF with other ALFs selleck chemicals revealed the presence of conserved regions within the sequence ( Fig. 1C). The phylogenetic relationship between Sc-ALF and other ALFs of decapod crustaceans was analyzed using the Neighbor-Joining (NJ) method ( Fig. 1D). Molecular phylogenetic tree based on amino acid sequences suggests that all the ALF members possess the same ancestral origin, which has subsequently

diverged at different phases of evolution. The tree could be broadly divided into two major groups, Group I included ALFs from shrimps and lobsters and Group II consisted of ALFs from crabs and crayfishes. The bootstrap distance tree calculated for the Sc-ALF clearly indicated that the Sc-ALF possessed great similarity to ALFs of other crabs ( Fig. 1D). The full-length cDNA of Sc-crustin was 433 bp in length, encoding 144 amino acids (Fig. 2A). The Sc-crustin cDNA encoded a polypeptide of 111 amino acids in the ORF with a putative see more signal peptide of 21 amino acid residues and a mature

protein of 90 amino acids (Fig. 2A). The calculated molecular mass of the mature protein was 10.24 kDa. The isoelectric Dapagliflozin point (pI) was estimated to be 8.76 as predicted by the PROTPARAM software. The full-length sequence was deposited in the NCBI GenBank under accession number HQ638025. Sequence comparison using BLAST algorithm showed that the deduced amino acid sequence of Sc-crustin possessed

an overall similarity of 81%, 62%, 73%, 56% and 39% to the crustins of S. paramamosain, P. trituberculatus, H. araneus, C. maenas and F. chinensis, respectively ( Table 2). The deduced amino acid sequence of Sc-crustin was found to be rich in amino acid residues cysteine (13.3%) and proline (11.1%). A WAP domain could be detected in the C-terminus of Sc-crustin. As described by Imjongjirak et al. [13] a conserved eight-cysteine residue region responsible for the formation of 4 disulfide core (4-DSC) could also be detected in the C5–C12 position ( Fig. 2A). The 12 cysteines in Sc-crustin ( Fig. 2A) are considered to be important for maintaining the tertiary structure of the peptide just as that reported in the case of shrimp crustins [23] and [24]. Peptide model of Sc-crustin created using SWISS-MODEL server indicated a random coiled structure, that is, with two possible β-sheets but no helices ( Fig. 2B). Multiple alignment performed for Sc-crustin with other crustins of decapods revealed the presence of conserved regions within the sequence ( Fig. 2C). In the present study, the BLAST homology search for the AMPs (Sc-ALF and Sc-crustin) from the brown mud crab, S. serrata showed maximum similarity to those from the green mud crab, S.

Induction of Cec2 (group III gene representative) mRNA was attenuated to 40% in MyD88 knockdown animals and to about 55% in IMD knockdown animals at 24 h post Ec challenge. At 6 h the reduction was not obvious with even elevated mRNA levels for IMD knockdown. Taken together, induction of Att1, Col1, Def2 (group I) and Def3 (group II) mRNAs by Ec challenge were weakened and nearly eliminated by 24 h in IMD knockdown animals while MyD88 knockdown affected Def3 (group II) and Cec2 (group III) induction. Similarly, induction of the five representative AMP genes by Ml challenge was examined in the MyD88 and IMD knockdown animals at 6 and 24 h post bacterial

injection ( Fig. 3C and D). Basically, the overall induction profiles had a similar tendency to the case of Ec challenge. Group I genes exhibited the dependence on IMD, which was Dolutegravir datasheet more conspicuous at 24 h post Ml challenge. Induction of Def3 (group II)

at 6 h was attenuated in both MyD88 and IMD knockdown animals as in the case of Ec challenge whereas at 24 h it was elevated more than three times in animals treated with MyD88 dsRNA, and it remained at a similar level to the control in IMD knockdown animals. Cec2 (group III) induction showed the dependence on MyD88 at both 6 and 24 h post Ml challenge, which was more obvious at 24 h. Thus, induction of Att1, Col1 and Def2 (group I) by Ml was weakened by IMD knockdown, while that of Cec2 (group III) was weakened by MyD88 knockdown. As for Def3 (group II), its expression RVX-208 seemed to be mediated by both MyD88 and find more IMD although the data at 24 h were obscure. In addition, Col1 and Def3 induction by Ml was enhanced at 24 h by MyD88 knockdown, and Def2 and Cec2 induction at 6 h after Ml injection was slightly elevated by MyD88 and IMD knockdown, respectively. The effects of IMD knockdown on the induction of group I genes

by Ec and Ml appeared more drastic at 24 h post challenge than at 6 h as mentioned above ( Fig. 1A–D). Zou et al. examined the microbial induction of several immune-related genes in the adult beetle using qRT-PCR [39]. According to their results, IMD is also inducible by microbial challenges as well as other immune-related components. We infer that on IMD knockdown background remaining IMD proteins in pupae may be consumed with time and may eventually be depleted by 24 h post bacterial challenge because of a loss of its de novo synthesis, which could cause more apparent knockdown effects of IMD on group I genes at 24 h. Other components of the pathway may be involved as well. AMP gene induction in knockdown animals by Sc challenge is shown in Fig. 3(E and F). Induction of Att1 and Col1 mRNAs were attenuated in MyD88 and IMD knockdown animals at both 6 and 24 h after Sc injection. Def2 induction was not suppressed at 6 h post Ml challenge by either of knockdown whereas it was weakened at 24 h by both dsRNA treatments.

The basic difference with “true” resin-based self-etch adhesives

is that the latter possess functional monomers with usually only one or two functional chemical groups with affinity to HAp. They thus provide individual monomers that upon polymerization become a polymer linked to HAp, in contrast to glass-ionomers that make use of an already existing (polyalkenoic-acid) Neratinib solubility dmso polymer with multiple functional groups that are attached to the polymer backbone and can “grab” Ca at different and remote sites. The additional chemical bonding provided by glass-ionomers and self-etch adhesives is believed to be advantageous in terms of bond durability [53] and [67]. In contrast, molecules such as phosphoric and maleic acid, but also functional monomers of self-etch adhesives such as Phenyl-P, will initially

bond to Ca of HAp, but then will readily de-bond. The negatively loaded phosphate ions (or carboxyl groups for carboxyl-based monomers/acids) will remove the positively loaded (and thus electro-statically attracted) Ca ions Apoptosis inhibitor from the surface, up to a certain depth depending on the application time. This results in a severe decalcification or “etching” effect, as it is best known in the case of phosphoric acid, which is used as an “etchant” in the “etch-and-rinse” approach. Because the calcium-phosphate/carboxylate bond originally formed at the enamel/dentin Adenosine surface is not stable, the bond will dissociate, leading to a typical etch pattern at enamel and a relatively deep (3–5 μm) hybrid layer at dentin that no longer contains any apatite

crystals. The pKa value of an acid is generally considered the major parameter that determines how its molecules interact with mineralized tissues [49] and [68]. However, this does not fully explain the mechanisms by which certain molecules adhere to tooth tissue, while others do not, but rather severely decalcify it [63] and [64]. For instance, 1 M oxalic acid (pK1 = 1.27, pK2 = 4.28) with a pH of 0.6 is more acidic than 10% maleic acid (pK1 = 1.94, pK2 = 6.23), which has a pH of 0.9. Nevertheless, oxalic acid chemically bonds to HAp, while maleic acid decalcifies it. In other words, it is not necessarily true that the lower the pH (the more acidic), the more the solution will demineralize enamel and dentin [63]. Keeping apatite at the interface is important to protect collagen and generate chemical interaction receptiveness. Dentinal collagen exposed by an etch-and-rinse procedure has been documented to be highly vulnerable to hydrolytic and enzymatic degradation processes [69], [70] and [71]. Actually, the fact that an etch-and-rinse hybrid layer can be demineralized, confirms the relatively permeable nature of the resin-impregnated collagen layer and perhaps its consequent instability in the long term.

04 L (90.7% of predicted), 2.84 L (86.9%), 2.30 L (86.8%), and 81.0% respectively.

TLC, RV, and RV/TLC were 4.80 L (93.8%), 1.76 L (112 %), and 36.7% (100%) respectively. DLCO was 14.64 mL/min/mmHg (88.9%) and DLCO/VA was 3.73/min/mmHg (84%). Bronchoalveolar lavage (BAL) (recovery rate; 63/150) revealed high total cellularity (36 x 104 cells/ml) consisting of 22% lymphocytes, 2% neutrophils, and 2% eosinophils. BAL lymphocytosis was suggestive of BAY 73-4506 manufacturer NSIP and transbronchial biopsies of the right lung (rS9) showed lymphocyte infiltration with no evidence of infection. However, the specimens obtained were insufficient to allow for a definitive diagnosis. The patient then underwent thoracoscopic biopsy. The main finding of the surgical biopsy specimen (rS9) was irregular interstitial fibrosis with mild chronic inflammation, which ranged

from the peripheral to the central part of lobule (Fig. 1c). The patchy distribution of fibrotic changes seen in some subpleural regions was similar to UIP, while the fibrotic process was temporally homogeneous. The lung architecture was relatively preserved and honeycombing selleck chemicals was not observed. These pathological findings were consisted with that of fibrotic NSIP, and centrilobular emphysema in the non-fibrotic lesion and focal intraluminal accumulation of macrophages suggested superimposed smoking effects (Fig. 1d, e). Clinical, radiological, and pathological information established the diagnosis of idiopathic NSIP. Although we planned to treat the patient with prednisolone plus an immunosuppressive agent, he refused the medication due to an improvement in his cough following the complete cessation of smoking after the surgical biopsy. Moreover, ground-glass opacity and reticular patterns on HRCT were found to have gradually improved without medication during the next 4 months (Fig. 1f) and KL-6 was reduced to

392 U/ml. No evidence of exacerbation was detected during the 15-month follow-up. Possible pathogenic factors implicated in smoking with interstitial fibrosis may include oxidative stress [4], decreased HDAC2 activity [5], VEGF expression [6], and the up-regulation of TNF-α [7]. However, the impact of the cessation of smoking during the clinical course of NSIP remains to be established. PFKL To the best of our knowledge, this is the first reported case of fibrotic NSIP that markedly improved without medication after the complete cessation of smoking, which suggested that smoking may be an etiological factor in some patients with NSIP. The association of emphysema with NSIP, as shown in our case, and differences in the morphological features on HRCT between non-smokers and smokers may support this hypothesis [3]. Differential diagnosis of this case includes DIP, combined pulmonary fibrosis and emphysema (CPFE), and smoking-related interstitial fibrosis (SRIF). DIP usually responds to corticosteroid therapy. However, some cases progress to fibrosis, despite treatment [8].

Water was Milli-Q (Millipore, USA). General solvents were from Merck. Young (1 month) and mature (6 month) leaves from I. paraguariensis were collected randomly from two areas: from a disturbed forest enriched with Maté plants, and from a homogeneous group of cultivars, exposed to sunlight (monoculture), with geographical coordinates 27°37′15″ south, 52°22′47″ west at 765 m altitude (Barão de Cotegipe, State of Grande do Sul). Harvesting was in the winter month, July 2009. The leaves were grouped in four clusters:

mature sun-exposed and shade-submitted leaves, young sun-exposed and shade-submitted leaves. These were kept without processing (in natura), or subjected to blanching/drying (as with “chimarrão”) or oxidation (as with black tea), Cilengitide yielding 12 samples ( Supplementary Table 1). Freshly harvested leaves were dried in an oven with air circulation at 30 °C for 24 h. Thereafter, they were exposed to flame (“sapeco”) at 180 °C for 5 min (residual moisture ∼ 15%) and, then, dried at 65 °C for 90 min (moisture ∼ 5%). The leaves

were submitted to dehydration for 2 h using an oven with air circulation at 30 °C, and manually rolled at room temperature (25 °C) for 5 min. The leaves were then transferred to aluminium trays and submitted to experimental conditions (26 °C and 80% relative humidity) for 3 h. Thereafter, find more they were dried at 70 °C for 120 min. The leaves were ground and a portion of 100 g of each was submitted to aqueous extraction (100 °C, 500 ml, x3). The extracts were combined and evaporated to a small volume. High molecular weight components were precipitated by addition to cold EtOH (x3 v/v), and separated by centrifugation (8.000 rpm

at 4 °C, 20 min). Ethanol-soluble fractions were concentrated under reduced pressure, and cAMP were then freeze-dried and stored in freezer. Monosaccharides and oligosaccharides were analysed using HPTLC, performed with silica gel 60G plates (Merck, Darmstadt, Germany). The samples were prepared in water at 2 mg/ml, with 5 μl being applied to the plate, which was developed with EtOAc:H2O:HOAc:HCOOH (9:2.3:1:1). The carbohydrates were stained by orcinol–H2SO4 at 100 °C (Sassaki, Souza, Cipriani, & Iacomini, 2008). Samples (100 μg/ml) in MeOH–H2O (1:1, v/v) containing LiCl 5 mM, were submitted to positive and negative atmospheric pressure ionisation (API), recorded in a triple quadrupole, Quattro LC (Waters), with nitrogen as nebuliser and desolvation gas. Offline analyses were performed by direct injection of the samples into the ESI-MS source, aided by a syringe-infusion pump at a flow rate of 10 μl/min. Second stage tandem-MS profiles were obtained by collision induced dissociation-mass spectrometry (CID-MS), using argon as collision gas. UPLC was used for quantification of carbohydrates, xanthines and phenolics. Calibration curves (R2 > 0.