Blood was collected in tubes containing K3EDTA and refrigerated until the hematological analysis (up to 2 h). The blood analyses were performed at least in triplicate. Red blood cells and platelets were also measured. Samples for the biochemical assay were collected in tubes with a coagulation enhancer and splitting gel (Vacuette, Greiner Bio-One) and immediately centrifuged www.selleckchem.com/products/MK-1775.html (3,000 × g, 10 min). The blood serum was aliquoted and stored in liquid nitrogen for future analyses.

The sera were analyzed using clinical kits for the following muscle injury markers and biochemical variables: ammonia, creatine kinase (CK), creatine kinase-MB (CK-MB), aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyltransferase (γGT), lactate

dehydrogenase (LDH), alkaline phosphatase (ALP), glucose, urea, creatinine, urate, total protein, albumin, bilirubin, globulins, and serum hemoglobin. ACP-196 No changes in plasma volume were detected during the experiment. Calculations and statistics The area under the curve (AUC) for the blood ammonia data for each individual in each treatment was determined using the equation AUC = Ai(Ti + 1 – Ti) + (1/2)(Ai + 1 – Ai)(Ti + 1 – Ti), where A denotes ammonia concentration (μmol/L) and T denotes time (min). The blood ammonia accumulation rate during the match was calculated by the difference between the ammonia concentrations before and approximately 1 minute after exercise divided by 6 minutes. The data are shown as the mean

and standard error. The data were normalized to the pre-exercise values. The intergroup statistical significance was calculated by a one-way analysis of variance (ANOVA), and the intragroup 5-FU cost significance was established by Student’s t-test. The data selleck products correlations were calculated using Pearson’s test. Significant differences were assumed at P < 0.05. Results Proteins and injury markers To ensure that the athletes were at similar training levels and had similar liver integrities, we measured the classic muscle and liver injury markers. The athletes of both groups had similar anthropometric values (Table 1). Despite the high levels of classic muscle injury markers, such as CK (EC 2.7.3.2) and LDH (EC 1.1.1.27), the concentrations of these enzymes in the blood did not change after the match. The liver injury markers ALP (EC 3.1.3.1) and γGT (EC 2.3.2.2) also remained stable in both groups. The same stability was observed with the less specific markers AST (EC 2.6.1.1) and ALT (EC 2.6.1.2) (Table 2). The amount of globulins in the blood increased in both groups after exercise, with an 11% increase in the RG and a 15% increase in the PG (Table 2). Table 1 Age and anthropometric measurements in Brazilian Jiu-Jitsu fighters assigned to the PG and RG   PG Range RG Range Age (years) 25.2 ± 0.4 21-28 26.2 ± 0.6 23-29 Weight (kg) 82.2 ± 1.8 70-103 79.2 ± 3.2 65-120 Height (cm) 177 ± 1.0 170-188 175 ± 1.

Patients with proteinuria (urine dipstick ≥2+), impaired creatinine clearance (<30 ml/min), or abnormal serum calcium levels (>2.75 or <2.08 mmol/l) at screening were not eligible for study participation. Patients with an ongoing infection (including dental infection) or planned oral surgery within 3 months of randomization were also excluded. Study medications All patients received an infusion of ZOL 5 mg over at

least 15 min on Day 1. Patients were randomized to one of three treatment groups. All www.selleckchem.com/products/lazertinib-yh25448-gns-1480.html oral study drugs were double-blinded with matching placebo capsules. In Group 1, 45 min prior to ZOL infusion, two capsules of acetaminophen 325 mg were administered orally. Subjects in this group continued to take two capsules of acetaminophen four times daily for the

next 3 days at home. In Group 2, 45 min prior to ZOL infusion, two capsules of immediate-release fluvastatin 40 mg were administered orally. Fluvastatin was administered only once, on Day 1, prior to ZOL infusion. Subjects in the fluvastatin group were provided with placebo capsules (matching acetaminophen) and took two capsules four times daily for the next 3 days at home. In Group 3, subjects received placebo 45 min prior to ZOL infusion and continued to take two capsules of placebo (matching acetaminophen) four times daily for the next 3 days at home. Patients in this study were also provided with calcium 600 mg plus vitamin D3 400 IU (one tablet twice daily). Open-label rescue medication (ibuprofen 200 mg Cyclooxygenase (COX) tablets DNA Damage inhibitor every 4 to 6 h, not to exceed eight tablets in a 24-h period) could be taken if the patient had an oral body temperature ≥38.9°C (102°F) and/or severe symptoms of fever, headache, myalgia, or arthralgia. Study personnel observed each patient ingest the first dose of study medication and receive the ZOL

infusion. Patient diaries and unused medication were used to assess compliance during the remainder of the study. Patients recorded the dose, date, and time of study and rescue medication use in diaries and returned used bottles and unused study and rescue medication at the final visit. Efficacy and safety variables The primary efficacy variable in this study was the proportion of patients who had a clinically significant increase in oral body temperature (≥1°C from baseline and ≥38.5°C overall) or used rescue medication at least once during the 3-day period following ZOL infusion. Oral body temperature was measured at baseline prior to ZOL infusion using a digital thermometer (Welch Allyn Sure Temp), which was provided to each patient for the duration of the study. Patients were GDC 941 trained to take two temperature assessments within 10 min of each other four times per day for 3 days. Temperature assessments were conducted after completing VAS and symptom questionnaires and prior to taking any oral study medication.

It is useful to compare the spectra from

the unknown complex to some known model complexes (assuming that there is selleck inhibitor evidence that the structure resembles that of the model complex) and then use Debye–Waller parameters obtained from the model complexes in the fits. This method works reasonably well, when the structure of the system being studied is well-modeled by inorganic complexes.   X-ray absorption spectroscopy studies of photosystem II One of the advantages of XAS is that one can potentially study the chemical events from each element which is involved in the reaction. In the OEC, Mn, Ca, and possibly Cl are the key elements we can focus on, in order to obtain https://www.selleckchem.com/products/bgj398-nvp-bgj398.html the mechanistic information during the catalytic cycle.

The XAS results, with emphasis on results from our laboratory, will be used to highlight the utility of the technique for the study of the Mn4Ca cluster in PS II. Mn XAS The geometric and electronic structural changes of the OEC have been studied intensively using Mn XAS. Figure 3 shows the Mn K-edge spectrum of each S-state of spinach PS II after deconvolution of the spectra obtained from consecutive flash illumination into pure S-state spectra, and their second derivative spectra (Messinger et al. 2001). Traditionally, the inflection point Cisplatin in vivo of the rising Mn K main edge (electron 1s to 4p transition) has been used as an indicator of the oxidation states in the field of XAS. The edge positions for each of the S-states have been quantitated by measuring the inflection

point energy (IPE), given by the zero-crossing of the second derivative. Extensive model compound studies have shown that, when Mn is oxidized by one electron in a set of Mn model compounds with similar ligands, the IPE shifts 1–2 eV to higher energy (Visser et Sinomenine al. 2001). Clear differences in absorption edge energy attributed to Mn oxidation were seen in the S0 → S1 and S1 → S2 transitions in the OEC, but the absorption edges for S2 and S3 did not show a significant difference. These results were taken to indicate the absence of Mn oxidation during the S2 → S3 transition, although different interpretation exists. However, one has to be aware that the edge position cannot be simply an indicator of only the oxidation state and it is problematic to conclude oxidation state changes based only on the XANES inflection point. Due to the size of the metal 4p orbital, this orbital overlaps with p orbitals of the ligands, either through σ- or π-bonding. Consequently, XANES is sensitive not only to the oxidation state but also to the ligand environment of the metal. Additionally, no definite theory is available for calculating main K-edge spectra for transition-metal complexes, owing to several factors that affect the metal p-density.

H. pylori genomes were extracted using genomic DNA isolation kits (Omega Biotek Inc).

Culture and identification of H. pylori were done by appropriate biochemical tests and amplification of 16S rDNA using species-specific primers Selection and identification the VNTR loci of H. pylori VNTR loci were selected from the MLVA database http://​minisatellites.​u-psud.​fr/​ASPSamp/​base_​ms/​bact.​php by buy Daporinad estimating the size of PCR products on agarose gels. The repeat sequence of loci ≥ 10 bp, consistency of repeat unit ≥ 90% and a minimum of two alleles in three reference strains of H. pylori (26695, HPAG1, J99) were selected for this research. The locations, copy numbers, sizes of the loci and the gene(s) involved are also listed in Table 1. PCR amplification A PCR reaction mixture (30 ml) containing 10 ng of DNA template, 0.5 mM of each primer, 1 unit of Taq DNA polymerase, 200 mM of dNTPs and 10 × PCR buffer (500 mM KCl, 100 mM TrisHCl (pH 8.3) 25 mM MgCl2) was utilized. Amplification was carried out in a DNA thermocycler (MJ Research PTC-225) with denaturation at 94°C for 8 min, followed by 30 cycles of denaturation at 94°C for 45 s, annealing

at 52°C for 45 s and elongation at 72°C for 1 min [26]. A 10-min elongation at 72°C was performed after MK-1775 mw the last cycle to ensure complete extension of the amplicons. Five μl of the PCR products were run on standard 3% agarose gels in 0.56TBE buffer at 8-10 V/cm. Gel lengths of

10 to 40 cm were used according to PCR product size and repeat unit size. Strains in which alleles had been precisely measured by re-sequencing or by direct comparison with a sequenced reference strain were used (In this study DNA from 26695, HPAG1 and J99 were used for this purpose). Multiple interspersed negative controls containing no DNA were included each time PCR was performed. PCR products of 202 strains on VNTR-2576 and VNTR-614 sites were sequenced directly with a Taq Dye Sinomenine Deoxy Terminator Cycle Sequencing Kit on an ABI 377 sequencer (Applied SB203580 in vitro Biosystems). Data analysis The number of repeat units in 12 VNTR loci were analyzed and inputted into BioNumerics version 5.1 software (Applied-Maths, Sint-Martens-Latem, Belgium), and gel images were obtained using the BioNumerics software package version 6.0 (Applied-Maths, Sint-Martens-Latem, Belgium) or using UVB gel image analysis. The number of repeat units in each locus was deduced by the amplicon size, flanking sequence length and repeat unit size.

Our results also seemed to support this hypothesis since both high bacterial production and specific bands were only observed in treatments VF and VFA. Stimulation of viral production was much more variable between lakes than between seasons and it was clearly higher in Lake Bourget. This suggests that environmental conditions encountered in the mesotrophic system might promote higher viral activity compared to more oligotrophic conditions. This hypothesis agrees with Lymer et al. [34] or Pradeep and Sime-Ngando [26] who observed, during a microcosm experiment, an enhancement of both viral abundance and FIC (frequency of infected cells) in P-enriched

samples as a result of nutrient stimulation of bacterial growth, which in turn enhanced viral activity. However, it is noteworthy

here that although check details phosphorus concentration was 2-fold higher in Lake Bourget than in Lake Annecy (Table 1), no significant difference was recorded in bacterial production between the two lakes (t test, P > 0.005). Some studies have suggested that nutrient availability may have an important influence on viral life strategies (e.g. [35, 36]). As lysogenic infection is considered the most favourable method of bacterial infection in water characterized by low bacterial abundance and primary production, this may also explain the relatively weak stimulation of viral production observed in Lake Annecy compared to Lake Bourget [32]. In Lake Annecy, and in contrast GDC-0449 mouse to viral production, the effects of flagellate presence on viral abundance seemed to be highly variable between the two periods (LA1 vs. LA2). This

variation revealed viral abundance stimulation in early-spring (LA1) and repression Ribose-5-phosphate isomerase in summer (LA2), for both treatments (VFA and VF). This result could suggest a direct grazing of flagellates on viruses during summer. Virivory by flagellates has been previously reported [37, 38] and according to Domaizon et al. [39], all flagellates do not act similarly because of large differences between taxon-specifc ingestion rates. During our study, selleck chemical heterotrophic flagellates were mainly represented by Oikomonas (45 and 48% during LA1 and LA2, respectively). Also, the grazing impact of flagellates on viruses has always been reported to be relatively low, resulting in < 4% loss [37, 38]. Hence, direct grazing of flagellates on viruses was unlikely to explain the repression of viral abundance in LA2. Other factors should be invoked [36] and would need further investigation. Effect of both flagellates and viruses on bacterial activity Higher bacterial production in both VF and VFA treatments than V suggested that grazers and viruses acted additively to sustain (directly or indirectly) bacterial activity in Lake Annecy and Lake Bourget.

J. Immunol. 2004, 173:437–445.PubMed 15. Volasertib chemical structure Bazzocchi C, Comazzi S, Santoni R, Bandi C, Genchi C, Mortarino M: Wolbachia surface protein (WSP) inhibits apoptosis in human neutrophils. Parasite Immunol 2007, 29:73–79.PubMedCrossRef 16. Vizioli J, Richman AM, Uttenweiler-Joseph S, Blass C, Bulet P: The defensin peptide of the malaria vector mosquito Anopheles gambiae : antimicrobial activities and expression in adult mosquitoes. Insect Biochem Mol Biol 2001, 31:241–8.PubMedCrossRef 17. O’Neill SL, et al.: In vitro cultivation of Wolbachia pipientis in an Aedes albopictus this website cell line. Insect Mol. Biol. 1997, 6:33–39.PubMedCrossRef

18. Turner JD, et al.: Wolbachia endosymbiotic bacteria of Brugia malayi mediate macrophage tolerance to TLR- and CD40-specific stimuli in a MyD88/TLR2-dependent manner. J. Immunol. 2006, 177:1240–1249.PubMed 19. Bazzocchi C, Ceciliani F, McCall JW, Ricci I, Genchi C, Bandi C: Antigenic role of the endosymbionts of filarial nematodes: IgG response against the Wolbachia surface protein in cats infected with Dirofilaria immitis . Proc. Biol. PLX3397 supplier Sci. London Ser. B 2000, 267:2511–2516.CrossRef 20. Müller HM, Dimopoulos G, Blass C, Kafatos FC: A hemocyte-like cell line established from the malaria vector Anopheles gambiae expresses six prophenoloxidase

genes . J. Biol. Chem. 1999, 274:11727–11735.PubMedCrossRef Methocarbamol 21. Pinto SB, et al.: Discovery of Plasmodium modulators by genome-wide analysis of circulating hemocytes in Anopheles gambiae. Proc Natl. Acad. Sci. U. S. A. 2009, 106:21270–21275.PubMedCrossRef 22. Dong YI, Aguilar R, Xi Z, Warr E, Mongin E, Dimopoulos G: Anopheles gambiae immune responses to human and rodent Plasmodium parasite species. PLoS Path 2006, 2:e52.CrossRef 23. Blagrove MC, Arias-Goeta C, Failloux AB, Sinkins SP: The Wolbachia strain wMel induces cytoplasmic incompatibility and

blocks dengue transmission in Aedes albopictus. Proc Natl. Acad. Sci. U. S. A., in press. Authors’ contributions SBP participated in the design of the study, carried out experimental work, data analysis and drafted the manuscript. MM carried out experimental work and data analysis. CB provided reagents and experimental support. ClB participated in the design of the study and helped draft the manuscript. SPS participated in the design of the study, provided reagents and drafted the manuscript. All authors read and approved the final manuscript. Competing interests The authors declare that they have no competing interests.”
“Background Wolbachia are a highly diverse group of intracellular, maternally inherited endosymbionts belonging to the α-Proteobacteria [1]. The bacteria infect a wide range of arthropods, including at least 65% of insect species [2–4], as well as filarial nematodes [5].

The patterns are shifted vertically for clarity. The annealed samples show the presence of NiO peaks. The reflexes of Ni are still observed and arise from the incomplete oxidation of the Ni supporting layer. The stars and tick marks denote the Au-Ni alloy and Au, respectively. From the above, it can be seen that metallic Ni still dominate the XRD spectrum, and it appears necessary to estimate the magnitude of oxidation of the nanostructures. For doing this, we make use of the data published in [33] which shows that Ni oxidation follows

a parabolic law in a wide range of temperature. Through extrapolation and taking into account the surface area of the 1D morphology involved (see calculation details in Additional file 1: S1), it can be selleck chemicals llc shown that sample 2 consists of 60% NiO while sample 3 is completely oxidized. Gemcitabine in vivo Using the same procedure, only a small fraction of oxide (0.37%) is calculated for the underlying Ni layer, which explains the dominance of the Ni peaks in the XRD patterns. The morphology

of the nanostructures obtained is shown in Figure 2. The non-annealed sample 1 (Figure 2a, b) shows solely Ni NTs that form via nucleation and growth at the pore walls because of the presence of an extremely thin Au layer (see the experimental section and our previous paper [32]). The judicious deposition time for Ni to obtain NT is 50 s. Figure 2 SEM images of non-annealed (sample 1) and annealed samples (samples 2 and 3). (a) Cross-sectional and (b) top BCKDHB views of the as-prepared Ni NT (non-annealed sample 1 inside AAO template). (c) Wall thickening after 25-min annealing (sample 2). (d) The complete closure of walls yielding NR morphology after 300-min annealing (sample 3). During annealing, the oxide layer nucleates and grows from the exposed inside walls and thickens in the direction of the inner-tube diameter. This suggests an outward SCH727965 diffusion

of the Ni species toward oxygen ions. On the non-exposed outside walls that are confined by the AAO template, no oxide growth is expected. A short annealing time leads to incomplete oxidation of the Ni NTs, resulting in the formation of an oxide scale supported on a remaining Ni layer (see also the XRD results above and Additional file 1: S1). This is the case of sample 2 (Figure 2c; 25-min annealing). For longer annealing time, complete closure of the NT, to finally give the NR morphology as shown in Figure 2d, is achieved because of the volume increase associated with NiO oxide formation. This is the case of sample 3 (300-min annealing). Figure 3 shows the CV curves of the NiO NTs and NiO NRs recorded using a potential window of 0.5 V (between 0.35 and 0.85 V) at various scan rates (5, 10, 25, 50, and 100 mV/s).

The hyaluronidases Erastin can be subdivided into three types [15]: 1) hyaluronate-4-glycanohydrolases (EC 3.2.1.35), that are present in mammalian spermatozoa, lysosomes and the venoms of various insects and snakes; 2) hyaluronate-3-glycanohydrolases (EC 3.2.1.36), that are produced by leeches and some hookworms and 3) bacterial hyaluronidases or hyaluronate lyases (EC 4.2.2.1 or EC 4.2.99.1). Commonly used hyaluronidases are the partially purified bovine and ovine testicular ones. In spite of such a wide employment of both HA and Hy, only a few studies

have been conducted to assess their possible combined effects, if any, on protechnological or probiotic bacteria. Based on the survey of Ardizzoni et al. (2011) [8], focused on the inhibitory effect of HA on a group of pathogenic bacteria and fungal strains, the aim of the present study was to evaluate the effects of HA on potential probiotic Lactic Acid Bacteria (LAB). Results and discussion LAB engraftment within human gut has been the main challenge of last decade. However, well standardized MLN0128 purchase procedures to achieve a long lasting engraftment

still lack. This study, has been focused upon HA- Hy – LAB interaction to promote bacterial engraftment and feeding in order to enhance and prolong their beneficial effects. Firstly, the antimicrobial effect of HA was evaluated by MIC test in MRS agar. Among strains listed in Table 1, no one proved to be inhibited by HA even at a concentration of 4 mg ml-1. pH values of HA dilutions ranged from 6.5 to 7.6, corresponding to an HA concentration of 4 and 0.0625 mg ml-1, respectively. Moreover, when Lactobacillus (Lb.) rhamnosus LbGG cells Progesterone were exposed, for 30 min, to different levels of HA (4–0.0625 mg ml-1) a slight increase (about 0.5 log CFU ml-1) in microbial counts was recorded (data

not shown). In other words, high molecular weight HA did not exert any antimicrobial activity when tested on several LAB strains, but, on Adavosertib contrary, it seemed to enhance the bacterial viability. Table 1 Strains used in this study and source of isolation Taxon Strain Source Reference Lb. rhamnosus LbGG American Type Culture Collection ATCC53103 Lb. casei 491 Provolone del Monaco cheese [16] Lb. casei 496 Provolone del Monaco cheese [16] Lb. pentosus OM13 Table olives [17] Lb. rhamnosus VT1 Parmigiano Reggiano cheese [18] Lb. rhamnosus RBM526 Parmigiano Reggiano cheese [18] Lb. rhamnosus RBT739 Parmigiano Reggiano cheese [18] St. macedonicus 67 Provolone del Monaco cheese [19] St. thermophilus 309 Provolone del Monaco cheese [19] St. thermophilus 247 Provolone del Monaco cheese [19] St.

In addition, there are a number of studies in the literature on the brittle-ductile transition phenomenon of silicon material in nano-scale machining or indentation. For instance, Tanaka et al. observed amorphous phase transformation of silicon in nano-machining and that stable shearing of the amorphous region is necessary KPT-8602 for ductile-mode machining [15]. Also, a numerical

study of surface residual stress distribution of silicon during nano-machining process is presented by Wang et al. [16]. Their MD simulation results revealed that higher hydrostatic pressure beneath the tool rake face induces more drastic phase transformation and thus generates more compressive surface residual stress. MD simulation see more is also capable of modeling chip formation, separation, and evolution mechanism. For instance, Ji et al. [17] studied the tool-chip stress distribution in nano-machining of copper, and

the results were compared to the existing models of conventional machining. Lin and Huang [18] studied nano-cutting process by MD simulation and proposed the innovative ‘combined Morse potential function and rigid tool space restrictions criterion’ as the chip separation criterion. It was used to establish the shape function of the FEM-MD combined model. Existing studies on MD simulation of nano-scale machining usually adopt defect-free monocrystalline structures as the work material [19]. The most popular ones have been monocrystal copper, aluminum, and silicon. Nevertheless, the vast majority of engineering materials exist in polycrystalline (instead of monocrystalline) forms. It is not difficult to understand that machining polycrystalline structures may yield different results compared with machining monocrystalline structures. Moreover, the grain size in polycrystalline structures is often a controlling factor for material properties

and material responses to deformation. It is important to investigate how it impacts the machining performance check details at nano/atomistic scale. In a preliminary study, Shi and Verma [20] constructed one polycrystalline copper structure, simulated nano-scale machining of the structure, and made a comparison with monocrystalline machining. It was discovered that for all cutting conditions simulated, the polycrystalline structure requires smaller cutting forces compared with the monocrystalline structure. This result might be expected as the KPT-330 clinical trial existence of grain boundary is usually regarded as defects, and thus, it reduces material strength.

9% of the total variation of microbial community structure, 9.6%

of detected functional genes involved in C cycling, and 9.4% of detected functional genes in N cycling in this study. After accounting for the effects of the CO2 treatment, the selected variables from plant and soil could significantly explain more than 42% of the total variances of microbial community structure. Our previous studies have demonstrated that increased C inputs at eCO2 stimulate microbial activity and regulate their composition [13, 25]. Consistently, our statistical analysis suggests that the biomass of N2-fixing legume species (BLP) and the number of plant functional groups selleck screening library (PFG) have significantly positive correlations with the atmospheric CO2 level. These strong correlations could arise because increased plant-derived substrates at eCO2 could fuel heterotrophic metabolism in soil [44]. Such a strong correlation with the biomass of N2-fixing legume species (BLP) may result in an increased amount of N derived from the JQEZ5 in vitro atmosphere. Therefore, significant increases in plant biomass were associated

with the significant increase in the abundance of nifH genes, but little effect was seen in soil N dynamics. Soil microbial community structure may be shaped by soil properties, such as pH and moisture [45]. For example, soil pH and moisture changed at eCO2 in the BioCON study [6, 46], and a significant correlation between the soil microbial community compositions and soil pH was observed with a survey of 88 soils

across North and South America [47]. In this study, soil N% at the depth of 0-10 cm (SN0-10) and 10–20 cm (SN10-20), soil C and N ratio at the Selleck Tozasertib depth of 10–20 cm (SCNR10-20), and soil pH (pH) were identified as the most important soil factors shaping microbial community structures. In addition, significant correlations were also observed between the plant and soil factors, such as positive correlations between pH and BBG, pH and PFG, SCNR10-20 and BBG, and negative correlations between SCNR10-20 and BLP. These results suggested that, in addition to direct effects of atmospheric CO2 on soil microbial C and N cycling, such as CO2 fixation, eCO2-induced indirect effects on plant and soil properties significantly Florfenicol impact the soil microbial community structure and modify their ecosystem functioning. The simultaneous enhances in the processes involved in CO2 fixation, C degradation, N fixations and partial denitrification could be the reason that no significant difference was detected in total soil C and N. Conclusions GeoChip was successfully used to illuminate the response of soil microbial communities to eCO2. The results showed that microbial C and N cycling were altered dramatically at eCO2, and the eCO2-induced effects, such as increased plant biomass and altered soil pH, may largely shape the soil microbial community structure and regulate their ecosystem functioning.