Partial Hydrolysis of Diphosphonate Ester During the Formation of Hybrid TiO2 Nanoparticles: Role of Acid Concentration.

The hydrolysis of the phosphonate ester linker during the synthesis of hybrid (organic-inorganic) TiO2 nanoparticles is important when forming porous hybrid organic-inorganic metal phosphonates. In the present work, a method was utilized to control the in-situ partial hydrolysis of diphosphonate ester in the presence of a titania precursor as a function of acid content, and its impact on the hybrid nanoparticles was assessed. Organodiphosphonate esters, and more specific, their hydrolysis degree during the formation of hybrid organic-inorganic metal oxide nanoparticles, are relatively under explored as linkers. Here, a detailed analysis on the hydrolysis of tetraethyl propylene diphosphonate ester (TEPD) as diphosphonate linker to produce hybrid TiO2 nanoparticles is discussed as a function of acid content. Quantitative solution NMR spectroscopy revealed that during the synthesis of TiO2 nanoparticles, an increase in acid concentration introduces a higher degree of partial hydrolysis of the TEPD linker into diverse acid/ester derivatives of TEPD. Increasing the HCl/Ti ratio from 1 to 3, resulted in an increase in degree of partial hydrolysis of the TEPD linker in solution from 4 % to 18.8 % under the applied conditions. As a result of the difference in partial hydrolysis, the linker-TiO2 bonding was altered. Upon subsequent drying of the colloidal TiO2 solution, different textures, at nanoscale and macroscopic scale, were obtained dependent on the HCl/Ti ratio and thus the degree of hydrolysis of TEPD. Understanding such linker-TiO2 nanoparticle surface dynamics is crucial for making hybrid organic-inorganic materials (i. e. (porous) metal phosphonates) employed in applications such as electronic/photonic devices, separation technology and heterogeneous catalysis.

Pterosaur melanosomes support signalling functions for early feathers.

Remarkably well-preserved soft tissues in Mesozoic fossils have yielded substantial insights into the evolution of feathers1. New evidence of branched feathers in pterosaurs suggests that feathers originated in the avemetatarsalian ancestor of pterosaurs and dinosaurs in the Early Triassic2, but the homology of these pterosaur structures with feathers is controversial3,4. Reports of pterosaur feathers with homogeneous ovoid melanosome geometries2,5 suggest that they exhibited limited variation in colour, supporting hypotheses that early feathers functioned primarily in thermoregulation6. Here we report the presence of diverse melanosome geometries in the skin and simple and branched feathers of a tapejarid pterosaur from the Early Cretaceous found in Brazil. The melanosomes form distinct populations in different feather types and the skin, a feature previously known only in theropod dinosaurs, including birds. These tissue-specific melanosome geometries in pterosaurs indicate that manipulation of feather colour-and thus functions of feathers in visual communication-has deep evolutionary origins. These features show that genetic regulation of melanosome chemistry and shape7-9 was active early in feather evolution.

X-ray Absorption (XRA): A New Technique for the Characterization of Granular Activated Carbons.

The X-ray absorption (XRA) method using digital image processing techniques is a reliable technique to determine the exhaustion degree of granular activated carbons (GACs). Using an innovative digital image processing technique, the identification of individual adsorbed molecules or ions in a GAC was possible. Adsorption isotherm models (Langmuir and Freundlich) were used to simulate the adsorption equilibrium data of Methylene Blue (MB), nickel, cobalt and iodine. Freundlich equation was found to have the highest value of R2 compared with Langmuir. The identification of distinctive patterns applying XRA for different adsorbed ions and molecules onto GAC was explored. It is demonstrated that unique XRA configurations for each adsorbed ion or molecule are found, as well as a proportional relationship between its incident energy (needed to achieve maximum photon attenuation) and the (effective) atomic number, the adsorbate mass and the molar or atomic mass of adsorbed molecule or ion. XRA method in combination with image histogram modifications was used to obtain a digital signature of adsorbed ions/molecules, giving distinct GSI values for each one in the used energy range. Probabilistic models prove that XRA results are within relationships between effective atomic number and photonic interaction probability, reinforcing the potentialities of XRA for monitoring (multi-)ion and/or molecule combinations on GAC using advanced digital image processing techniques. It was proved that the proposed approach could assess different adsorbed ions/molecules onto GACs in water purification systems.

Fenton-Mediated Biodegradation of Chlorendic Acid - A Highly Chlorinated Organic Pollutant - By Fungi Isolated From a Polluted Site.

Chlorendic acid is a recalcitrant, highly chlorinated organic pollutant for which no microbial degrader has yet been identified. To address this knowledge gap, fungi were isolated from bulk soil, rhizosphere, and roots of the common bent (Agrostis capillaris) and the hybrid poplar [Populus deltoides × (Populus trichocarpa × P. deltoides) cv. Grimminge], both of which grow on a chlorendic acid polluted site in Belgium. Isolates were taxonomically identified and phenotypically screened for chlorendic acid degradation. Several fungal isolates could degrade chlorendic acid in liquid media up to 45%. The chlorendic acid degrading fungal isolates produced higher levels of hydroxyl radicals when exposed to the pollutant when compared to non-exposed controls, suggesting that the oxidative degradation of chlorendic acid occurs through production of Fenton-mediated hydroxyl radicals. In addition, the isolated Ascomycete Penicillium sp. 1D-2a degraded 58% of the original chlorendic acid concentration in the soil after 28 days. This study demonstrates that the presence of fungi in a chlorendic acid polluted soil can degrade this highly chlorinated organic pollutant. These results indicate that recalcitrant, seemingly non-biologically degradable organic pollutants, such as chlorendic acid, can be remediated by using bioremediation, which opens new perspectives for in situ bioremediation.

X-ray absorption as an alternative method to determine the exhausting degree of activated carbon layers in water treatment system for medical services.

Analytical methods based on X-Ray radiation proved to be reliable and sensitive techniques to study activated carbons (ACs). An X-Ray absorption analysis based on digital radiographic images (XRA) is applied for the determination of the exhaustion degree of granular activated carbon (GAC) used in a water purification system for hemodialysis. XRA-method demonstrated the possibility to determine the exhaustion degree at different layers of the GAC filter. The results of the XRA-method were successfully correlated with X-Ray Fluorescence (XRF), TGA, Elemental analysis (EA), SEM, TD-GC/MS, ATR-FTIR, X Ray Diffraction (XRD) and Nuclear Magnetic Resonance relaxometry (NMR) analyses. It was demonstrated that the XRA-method is a fast and reliable analytical tool to give indirect information on the exhaustion degree of GAC at different layers. It is also demonstrated that XRA results can be correlated with the results of the other analytical techniques, rather dealing with the composition and morphology of GAC at different layers.

Biodegradation of polycyclic aromatic hydrocarbons by native Ganoderma sp. strains: identification of metabolites and proposed degradation pathways.

Since polycyclic aromatic hydrocarbons (PAHs) are mutagenic, teratogenic, and carcinogenic, they are of considerable environmental concern. A biotechnological approach to remove such compounds from polluted ecosystems could be based on the use of white-rot fungi (WRF). The potential of well-adapted indigenous Ganoderma strains to degrade PAHs remains underexplored. Seven native Ganoderma sp. strains with capacity to produce high levels of laccase enzymes and to degrade synthetic dyes were investigated for their degradation potential of PAHs. The crude enzymatic extracts produced by Ganoderma strains differentially degraded the PAHs assayed (naphthalene 34-73%, phenanthrene 9-67%, fluorene 11-64%). Ganoderma sp. UH-M was the most promising strain for the degradation of PAHs without the addition of redox mediators. The PAH oxidation performed by the extracellular enzymes produced more polar and soluble metabolites such as benzoic acid, catechol, phthalic and protocatechuic acids, allowing us to propose degradation pathways of these PAHs. This is the first study in which breakdown intermediates and degradation pathways of PAHs by a native strain of Ganoderma genus were determined. The treatment of PAHs with the biomass of this fungal strain enhanced the degradation of the three PAHs. The laccase enzymes played an important role in the degradation of these compounds; however, the role of peroxidases cannot be excluded. Ganoderma sp. UH-M is a promising candidate for the bioremediation of ecosystems polluted with PAHs.

Body surface area-based versus concentration-based intraperitoneal perioperative chemotherapy in a rat model of colorectal peritoneal surface malignancy: pharmacologic guidance towards standardization.

Worldwide, cytoreductive surgery (CRS) and hyperthermic intraperitoneal perioperative chemotherapy (HIPEC) are used in current clinical practice for colorectal peritoneal surface malignancy (PSM) treatment. Although, there is an acknowledged standardization regarding the CRS, we are still lacking a much-needed standardization amongst the various intraperitoneal (IP) chemotherapy protocols, including the HIPEC dosing regimen. We should rely on pharmacologic evidence building towards such a standardization. The current IP chemotherapy dosing regimens can be divided into body surface area (BSA)-based and concentration-based protocols. A preclinical animal study was designed to evaluate pharmacologic advantage (PA), efficacy and survival. WAG/Rij rats were IP injected with the rat colonic carcinoma cell line CC-531. Animals were randomized into three groups: CRS alone or CRS combined with oxaliplatin-based HIPEC (either BSA- or concentration-based). There was no difference in PA between the two groups (p=0.283). Platinum concentration in the tumor nodule was significantly higher in the concentration-based group (p<0.001). Median survival did not differ between the treatment groups (p<0.250). This preclinical study, in contrast to previous thinking, clearly demonstrates that the PA does not provide any information about the true efficacy of the drug and emphasizes the importance of the tumor nodule as pharmacologic endpoint.

Body surface area-based vs concentration-based perioperative intraperitoneal chemotherapy after optimal cytoreductive surgery in colorectal peritoneal surface malignancy treatment: COBOX trial.

BACKGROUND AND OBJECTIVES: Cytoreductive surgery (CRS) and hyperthermic intraperitoneal perioperative chemotherapy (HIPEC) are the standard of care for patients diagnosed with colorectal peritoneal surface malignancy (PSM). Despite a clearly defined standardization of CRS, a large variety of HIPEC modalities are still used in clinical practice. METHODS: Body surface area (BSA)- and concentration-based HIPEC protocols were clinically and pharmacologically evaluated in a randomized phase III clinical pilot trial. Oxaliplatin dose was 460 mg/m 2 (BSA-based) in 2 L/m 2 carrier solution (concentration-based). Platinum quantification was performed using a validated inductively coupled plasma mass spectrometry method. Three-month morbidity, mortality, and health-related quality of life (HRQOL) were assessed. RESULTS: Thirty-one patients were randomized to either BSA- or concentration-based HIPEC. Toxicity and efficacy were higher (P < 0.001) in patients receiving concentration-based HIPEC. There was no difference in pharmacologic advantage between the two groups. A higher drug concentration in the tumor nodule at the end of HIPEC was found in the HIPEC-concentration group. There was no difference in major morbidity and mortality between the treatment groups. HRQOL was decreased 3 months postoperatively in the HIPEC-concentration group. CONCLUSION: Concentration-based chemotherapy delivers the drug in the most standardized way to the tumor nodule, resulting in increasing drug concentrations in the tumor nodule without increasing major morbidity.

In vivo Toxicity Assessment of Silver Nanoparticles in Homeostatic versus Regenerating Planarians.

Silver nanoparticles (AgNPs) belong to the most commercialized nanomaterials, used in both consumer products and medical applications. Despite its omnipresence, in-depth knowledge on the potential toxicity of nanosilver is still lacking, especially for developing organisms. Research on vertebrates is limited due to ethical concerns, and planarians are an ideal invertebrate model to study the effects of AgNPs on stem cells and developing tissues in vivo, as regeneration mimics development by triggering massive stem cell proliferation. Our results revealed a strong interference of AgNPs with tissue- and neuroregeneration which was related to an altered stem cell cycle. The presence of a PVP-coating significantly influenced toxicity outcomes, leading to elevated DNA-damage and decreased stem cell proliferation. Non-coated AgNPs had an inhibiting effect on stem cell and early progeny numbers. Overall, regenerating tissues were more sensitive to AgNP toxicity, and careful handling and appropriate decision making is needed in AgNP applications for healing and developing tissues. We emphasize on the importance of AgNP characterization, as we showed that changes in physicochemical properties influence toxicity.

Surface Chemistry of Oil-Filled Organic Nanoparticle Coated Papers Analyzed Using Micro-Raman Mapping.

Raman spectroscopy and micro-Raman mapping have been used to study the distribution of different chemical components at the surface of coated papers. The paper coatings contain organic nanoparticles with a structure of poly(styrene- co-maleimide) and encapsulated vegetable oils. Raman spectroscopy is able to differentiate between various types of oil, i.e., polyunsaturated, monounsaturated, or saturated, and indicates that the degree of imidization and reactivity of the oil (amount of free oil) complement each other. The surface mapping over large areas (5 × 5 mm2) illustrates good homogeneity of the coating layer and even surface coverage. The imide and oil are homogeneously distributed within the coating itself without a tendency for agglomeration. The covered areas of imide and oil mostly overlap for polyunsaturated oils, while larger amounts of oil occur outside the imide zones for monounsaturated and saturated oils. The latter indicates that the oil is partly "free" within the coating and acts as a continuous binder phase. The surface mapping over smaller areas (1 × 1 mm2) shows the coating and cellulose covered areas are complementary. The surface maps confirm that interaction between the coating and paper substrate happens through hydrogen bonding. Heterogeneities in the coating are due to the presence of remaining ammonolyzed maleic anhydride precursors forming amic acid moieties. The organic phase, oil phase, and cellulose substrate can also be differentiated by principal component analysis of the surface maps.

Adsorption and photocatalytic removal of Ibuprofen by activated carbon impregnated with TiO2 by UV-Vis monitoring.

The removal of Ibuprofen was investigated by activated carbon impregnated with TiO2. Emphasis was given on the effect of different parameters, such as composite type, initial Ibuprofen concentration (5-25 mg/L), temperature (22-28 °C) and pH (acidic and alkaline solution). The experiment was carried out in a self-made tubular flow reactor, with one 15 W monochromatic UV lamp (254 nm). The composite AC90T10 gives the highest removal degree of 92% of Ibuprofen solution under UV light within 4 h, due to synergy of adsorption and photodegradation. It was found that weight ratio of composite/Ibuprofen has limited effect on the removal degree within the concentration range (5-25 mg/L), but reaction time under UV light (4 h) and pH (acidic solution) are very important. The kinetic experimental data obtained at pH 4.3 at 25 °C on different composites were fitted to pseudo-first, pseudo-second and Elovich models, obtaining a high accuracy based on R2 values. From the results, composites of granular activated carbon and TiO2 can enhance removal of Ibuprofen effectively, making recycle process much easier and less costly, which can be a promising method in future water treatment.

The Sycamore Maple Bacterial Culture Collection From a TNT Polluted Site Shows Novel Plant-Growth Promoting and Explosives Degrading Bacteria.

Military activities have worldwide introduced toxic explosives into the environment with considerable effects on soil and plant-associated microbiota. Fortunately, these microorganisms, and their collective metabolic activities, can be harnessed for site restoration via in situ phytoremediation. We characterized the bacterial communities inhabiting the bulk soil and rhizosphere of sycamore maple (Acer pseudoplatanus) in two chronically 2,4,6-trinitrotoluene (TNT) polluted soils. Three hundred strains were isolated, purified and characterized, a majority of which showed multiple plant growth promoting (PGP) traits. Several isolates showed high nitroreductase enzyme activity and concurrent TNT-transformation. A 12-member bacterial consortium, comprising selected TNT-detoxifying and rhizobacterial strains, significantly enhanced TNT removal from soil compared to non-inoculated plants, increased root and shoot weight, and the plants were less stressed than the un-inoculated plants as estimated by the responses of antioxidative enzymes. The sycamore maple tree (SYCAM) culture collection is a significant resource of plant-associated strains with multiple PGP and catalytic properties, available for further genetic and phenotypic discovery and use in field applications.

Links Between Heathland Fungal Biomass Mineralization, Melanization, and Hydrophobicity.

Comprehending the decomposition process is crucial for our understanding of the mechanisms of carbon (C) sequestration in soils. The decomposition of plant biomass has been extensively studied. It revealed that extrinsic biomass properties that restrict its access to decomposers influence decomposition more than intrinsic ones that are only related to its chemical structure. Fungal biomass has been much less investigated, even though it contributes to a large extent to soil organic matter, and is characterized by specific biochemical properties. In this study, we investigated the extent to which decomposition of heathland fungal biomass was affected by its hydrophobicity (extrinsic property) and melanin content (intrinsic property). We hypothesized that, as for plant biomass, hydrophobicity would have a greater impact on decomposition than melanin content. Mineralization was determined as the mineralization of soil organic carbon (SOC) into CO2 by headspace GC/MS after inoculation by a heathland soil microbial community. Results show that decomposition was not affected by hydrophobicity, but was negatively correlated with melanin content. We argue that it may indicate that either melanin content is both an intrinsic and extrinsic property, or that some soil decomposers evolved the ability to use surfactants to access to hydrophobic biomass. In the latter case, biomass hydrophobicity should not be considered as a crucial extrinsic factor. We also explored the ecology of decomposition, melanin content, and hydrophobicity, among heathland soil fungal guilds. Ascomycete black yeasts had the highest melanin content, and hyaline Basidiomycete yeasts the lowest. Hydrophobicity was an all-or-nothing trait, with most isolates being hydrophobic.

A validated inductively coupled plasma mass spectrometry (ICP-MS) method for the quantification of total platinum content in plasma, plasma ultrafiltrate, urine and peritoneal fluid.

Oxaliplatin is a platinum (Pt)1 containing antineoplastic agent that is applied in current clinical practice for the treatment of colon and appendiceal neoplasms. A fully validated, highly sensitive, high throughput inductively coupled plasma mass spectrometry (ICP-MS) method is provided to quantify the total Pt content in plasma, plasma ultrafiltrate, urine and peritoneal fluid. In this ICP-MS approach, the only step of sample preparation is a 1000-fold dilution in 0.5% nitric acid, allowing the analysis of 17 samples per hour. Detection of Pt was achieved over a linear range of 0.01-100 ng/mL. The limit of quantification was 18.0 ng/mL Pt in plasma, 8.0 ng/mL in ultrafiltrate and 6.1 ng/mL in urine and peritoneal fluid. The ICP-MS method was further validated for inter-and intraday precision and accuracy (≤15%), recovery, robustness and stability. Short-term storage of the biofluids, for 14 days, can be performed at -4 °C, -24 °C and -80 °C. As to long-term stability, up to 5 months, storage at -80 °C is encouraged. Furthermore, a timeline assessing the total and unbound Pt fraction in plasma and ultrafiltrate over a period of 45 h is provided. Following an incubation period of 5 h at 37 °C, 19-21% of Pt was recovered in the ultrafiltrate, emphasizing the extensive and rapid binding of oxaliplatin-derived Pt to plasma proteins. The described method can easily be implemented in a routine setting for pharmacokinetic studies in patients treated with oxaliplatin-based hyperthermic intraperitoneal perioperative chemotherapy.

Physicochemical characterizations of functional hybrid liposomal nanocarriers formed using photo-sensitive lipids.

With recent advances in the field of diagnostics and theranostics, liposomal technology has secured a fortified position as a potential nanocarrier. Specifically, radiation/photo-sensitive liposomes containing photo-polymerizable cross-linking lipids are intriguing as they can impart the vesicles with highly interesting properties such as response to stimulus and improved shell stability. In this work, 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphoethanolamine (DTPE) is used as a photo-polymerizable lipid to form functional hybrid-liposomes as it can form intermolecular cross-linking through the diacetylenic groups. Hybrid-liposomes were formulated using mixtures of DTPE and saturated lipids of different chain lengths (dipalmitoylphosphatidylcholine (DPPC) and dimirystoilphosphatidylcholine (DMPC)) at different molar ratios. The physico-chemical characteristics of the liposomes has been studied before and after UV irradiation using a combination of techniques: DSC, QCM-D and solid-state NMR. The results signify the importance of a subtle modification in alkyl chain length on the phase behavior of the hybrid-liposomes and on the degree of crosslinking in the shell.

Fully quantitative description of hybrid TiO2 nanoparticles by means of solid state (31)P NMR.

For the first time, an absolute quantification of hybrid materials obtained from the reaction of phenylphosphonic acid (PPA) with TiO2 nanoparticles under different reaction conditions is reported. Next to the amount of PPA involved in grafting to the TiO2 nanoparticles, also the PPA included in titaniumphenylphosphonate crystallites is described quantitatively. The quantitative analysis is based on solid state (31)P MAS NMR and is further applied to evaluate the stability of the resulting hybrid materials towards hydrolysis and organic solvent exposure.

A novel acoustic approach for the characterization of granular activated carbons used in the rum production.

Acoustic analysis and sound patterns recognition techniques have been widely used in many branches of science, however; almost none focused on the characterization of granular activated carbon. A new methodology has been developed in order to characterize activated carbon based on the dynamic analysis in audible spectra of the sound's relative amplitude power produced by water flooded on granular activated carbon. A home-build recording set-up and management of acoustic measurements have been presented and correlated with the results of porous structure of carbons characterized by N2 adsorption. Five samples of granular activated carbons used in the rum production of different exhausted level have been evaluated by both methods. Parameters as the BET surface area and total pore volume showed a satisfactory correlation with acoustic measurement data when the signal is processed at 1000Hz. Three frequencies components of the produced sound were analyzed and related with the porous characteristics. The found relationship gives the possibility to predict and calculate textural parameters of granular activated carbons applying the acoustic technique. This methodology approach opens possibilities in using acoustic experiments for the characterization of high-porosity materials and to determine their exhausted level.

Ectomycorrhizal fungi decompose soil organic matter using oxidative mechanisms adapted from saprotrophic ancestors.

Ectomycorrhizal fungi are thought to have a key role in mobilizing organic nitrogen that is trapped in soil organic matter (SOM). However, the extent to which ectomycorrhizal fungi decompose SOM and the mechanism by which they do so remain unclear, considering that they have lost many genes encoding lignocellulose-degrading enzymes that are present in their saprotrophic ancestors. Spectroscopic analyses and transcriptome profiling were used to examine the mechanisms by which five species of ectomycorrhizal fungi, representing at least four origins of symbiosis, decompose SOM extracted from forest soils. In the presence of glucose and when acquiring nitrogen, all species converted the organic matter in the SOM extract using oxidative mechanisms. The transcriptome expressed during oxidative decomposition has diverged over evolutionary time. Each species expressed a different set of transcripts encoding proteins associated with oxidation of lignocellulose by saprotrophic fungi. The decomposition 'toolbox' has diverged through differences in the regulation of orthologous genes, the formation of new genes by gene duplications, and the recruitment of genes from diverse but functionally similar enzyme families. The capacity to oxidize SOM appears to be common among ectomycorrhizal fungi. We propose that the ancestral decay mechanisms used primarily to obtain carbon have been adapted in symbiosis to scavenge nutrients instead.

Rapeseed and Raspberry Seed Cakes as Inexpensive Raw Materials in the Production of Activated Carbon by Physical Activation: Effect of Activation Conditions on Textural and Phenol Adsorption Characteristics.

The production of activated carbons (ACs) from rapeseed cake and raspberry seed cake using slow pyrolysis followed by physical activation of the obtained solid residues is the topic of this study. The effect of activation temperature (850, 900 and 950 °C), activation time (30, 60, 90 and 120 min) and agent (steam and CO2) on the textural characteristics of the ACs is investigated by N2 adsorption. In general, higher activation temperatures and longer activation times increase the BET specific surface area and the porosity of the ACs, regardless of the activation agent or raw material. Steam is more reactive than CO2 in terms of pore development, especially in the case of raspberry seed cake. The performance of the ACs in liquid adsorption is evaluated by batch phenol adsorption tests. Experimental data are best fitted by the Freundlich isotherm model. Based on total yield, textural characteristics and phenol adsorption, steam activation at 900 °C for 90 min and CO2 activation at 900 °C for 120 min are found as the best activation conditions. Raspberry seed cake turns out to be a better raw material than rapeseed cake. Moreover, AC from raspberry seed cake produced by steam activation at 900 °C for 90 min performs as well as commercial AC (Norit GAC 1240) in phenol adsorption. The adsorption kinetics of the selected ACs are best fitted by the pseudo-second-order model.

Ectomycorrhizal Fungal Protein Degradation Ability Predicted by Soil Organic Nitrogen Availability.

In temperate and boreal forest ecosystems, nitrogen (N) limitation of tree metabolism is alleviated by ectomycorrhizal (ECM) fungi. As forest soils age, the primary source of N in soil switches from inorganic (NH4 (+) and NO3 (-)) to organic (mostly proteins). It has been hypothesized that ECM fungi adapt to the most common N source in their environment, which implies that fungi growing in older forests would have greater protein degradation abilities. Moreover, recent results for a model ECM fungal species suggest that organic N uptake requires a glucose supply. To test the generality of these hypotheses, we screened 55 strains of 13 Suillus species with different ecological preferences for their in vitro protein degradation abilities. Suillus species preferentially occurring in mature forests, where soil contains more organic matter, had significantly higher protease activity than those from young forests with low-organic-matter soils or species indifferent to forest age. Within species, the protease activities of ecotypes from soils with high or low soil organic N content did not differ significantly, suggesting resource partitioning between mineral and organic soil layers. The secreted protease mixtures were strongly dominated by aspartic peptidases. Glucose addition had variable effects on secreted protease activity; in some species, it triggered activity, but in others, activity was repressed at high concentrations. Collectively, our results indicate that protease activity, a key ectomycorrhizal functional trait, is positively related to environmental N source availability but is also influenced by additional factors, such as carbon availability.