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Characterizing the inorganic phase of biochar, beyond determining element concentration, is needed for appropriate application of these materials because mineral forms also influence element availability and behavior. Inorganics in 13 biochars (produced from Poultry litter, switchgrass, and different types of wood) were characterized by proximate analysis, chemical analysis, powder X-ray diffraction (XRD), and scanning electron microscopy with energy-dispersive X-ray (SEM-EDX) spectroscopy. Principal component analysis (PCA) was used to compare biochars and characterize associations between elements. The biochars were produced using commercial-scale reactors and represent materials with properties relevant to field application. Bulk inorganic concentration and composition were responsible for differentiating biochars after PCA of chemical data. In comparison, differentiation based on PCA of diffractogram fingerprints was more nuanced. Here, contributions from cellulose and turbostratic crystalline C influenced separation between samples. It was also sensitive to mineral forms of Ca (whewellite and calcite). Differences in crystalline C and Ca minerals separated two biochars generated from the same willow feedstock using the same pyrolysis conditions at different temperatures. PCA of 606 SEM-EDX point scans revealed that inorganics belong to four main clusters containing Ca, Fe, [Al, Si], and [Cl, K, Mg, Na, P, S] consistent with XRD identification of calcite, magnetic Fe-oxide, silicates, and sylvite. It further suggested that amorphous P-containing minerals associated with Ca (not identified through XRD) were constituents of willow and poultry litter-derived biochars. However, unlike PCA of XRD, it was not able to differentiate the two biochars derived from willow. The three analysis methods provided different perspectives on the properties of the biochar inorganic phase. Combining information from multiple methods is needed to better understand the inorganic composition of biochars., Competing Interests: The authors declare no competing financial interest.

Indoor exposures to metals arise from a wide variety of indoor and outdoor sources. This study investigates the impact of humid indoor conditions on the bioaccessibility of Zn in dust, and the transformation of Zn species during weathering. House dust samples were subjected to an oxygenated, highly humid atmosphere in a closed chamber for 4 to 5 months. Zinc bioaccessibility before and after the experiment was determined using a simulated gastric acid extraction. Bulk and micro X-ray absorption structure (XAS) spectroscopy was used to speciate Zn in dust. Exposure to humid conditions led to a significant increase in Zn bioaccessibility in all samples, which was due to a redistribution of Zn from inorganic forms toward the organic pools such as Zn adsorbed on humates. ZnO readily dissolved under humid conditions, whereas ZnS persisted in the dust. Elevated humidity in indoor microenvironments may sustain higher Zn bioaccessibility in settled dust compared to drier conditions, and part of this change may be related to fungal growth in humid dust. These results help to explain the greater bioaccessibility of certain metals in house dust compared to soils.

PACE4 plays an important role in the progression of prostate cancer and is an attractive target for the development of novel inhibitor-based tumor therapies. We previously reported the design and synthesis of a novel, potent, and relatively selective PACE4 inhibitor known as a Multi-Leu (ML) peptide. In the present work, we examined the ML peptide through detailed structure-activity relationship studies. A variety of ML-peptide analogues modified at the P8-P5 positions with leucine isomers (Nle, DLeu, and DNle) or substituted at the P1 position with arginine mimetics were tested for their inhibitory activity, specificity, stability, and antiproliferative effect. By incorporating d isomers at the P8 position or a decarboxylated arginine mimetic, we obtained analogues with an improved stability profile and excellent antiproliferative properties. The DLeu or DNle residue also has improved specificity toward PACE4, whereas specificity was reduced for a peptide modified with the arginine mimetic, such as 4-amidinobenzylamide.

Proprotein convertases (PCs) are crucial in the processing and entry of viral or bacterial protein precursors and confer increased infectivity of pathogens bearing a PC activation site, which results in increased symptom severity and lethality. Previously, we developed a nanomolar peptide inhibitor of PCs to prevent PC activation of infectious agents. Herein, we describe a peptidomimetic approach that increases the stability of this inhibitor for use in vivo to prevent systemic infections and cellular damage, such as that caused by influenza H5N1 and Shiga toxin. The addition of azaβ(3)-amino acids to both termini of the peptide successfully prevented influenza hemagglutinin 5 fusogenicity and Shiga toxin Vero toxicity in cell-based assays. The results from a cell-based model using stable shRNA-induced proprotein convertase knockdown indicate that only furin is the major proprotein convertase required for HA5 cleavage.

High-throughput screening of Tranzyme Pharma's proprietary macrocycle library using the aequorin Ca2+-bioluminescence assay against the human ghrelin receptor (GRLN) led to the discovery of novel agonists against this G-protein coupled receptor. Early hits such as 1 (Ki=86 nM, EC50=134 nM) though potent in vitro displayed poor pharmacokinetic properties that required optimization. While such macrocycles are not fully rule-of-five compliant, principally due to their molecular weight and clogP, optimization of their pharmacokinetic properties proved feasible largely through conformational rigidification. Extensive SAR led to the identification of 2 (Ki=16 nM, EC50=29 nM), also known as ulimorelin or TZP-101, which has progressed to phase III human clinical trials for the treatment of postoperative ileus. X-ray structure and detailed NMR studies indicated a rigid peptidomimetic portion in 2 that is best defined as a nonideal type-I' β-turn. Compound 2 is 24% orally bioavailable in both rats and monkeys. Despite its potency, in vitro and in gastric emptying studies, 2 did not induce growth hormone (GH) release in rats, thus demarcating the GH versus GI pharmacology of GRLN.

X-ray absorption fine-structure (XAFS) spectroscopy, micro-X-ray fluorescence (μXRF), and micro-X-ray diffraction (μXRD) were used to determine the speciation of Pb in house dust samples from four Canadian urban homes having elevated Pb concentrations (>1000 mg Pb kg(-1)). Linear combination fitting of the XAFS data, supported by μXRF and μXRD, shows that Pb is complexed in a variety of molecular environments, associated with both the inorganic and organic fractions of the dust samples. The inorganic species of lead identified were as follows: Pb metal, Pb carbonate, Pb hydroxyl carbonate, Pb oxide, and Pb adsorbed to iron oxyhydroxides. Pb carbonate and/or Pb hydroxyl carbonate occurred in all four dust samples and accounted for 28 to 75% of total Pb. Pb citrate and Pb bound to humate were the organic species identified. The results of this study demonstrate the ability of XAFS to identify Pb speciation in house dust and show the potential to identify Pb sources from new homes versus older homes. Understanding Pb speciation and how it influences bioaccessibility is important for human health risk assessment and risk management decisions which aim to improve indoor environmental health.

Vacuum samples were collected from 1025 randomly selected urban Canadian homes to investigate bioaccessible Pb (Pb(S)) concentrations in settled house dust. Results indicate a polymodal frequency distribution, consisting of three lognormally distributed subpopulations defined as "urban background" (geomean 58 μg g(-1)), "elevated" (geomean 447 μg g(-1)), and "anomalous" (geomean 1730 μg g(-1)). Dust Pb(S) concentrations in 924 homes (90%) fall into the "urban background" category. The elevated and anomalous subpopulations predominantly consist of older homes located in central core areas of cities. The influence of house age is evidenced by a moderate correlation between house age and dust Pb(S) content (R(2) = 0.34; n = 1025; p < 0.01), but it is notable that more than 10% of homes in the elevated/anomalous category were built after 1980. Conversely, the benefit of home remediation is evidenced by the large number of homes (33%) in the background category that were built before 1960. The dominant dust Pb species determined using X-ray Absorption Spectroscopy were as follows: Pb carbonate, Pb hydroxyl carbonate, Pb sulfate, Pb chromate, Pb oxide, Pb citrate, Pb metal, Pb adsorbed to Fe- and Al-oxyhydroxides, and Pb adsorbed to humate. Pb bioaccessibility estimated from solid phase speciation predicts Pb bioaccessibility measured using a simulated gastric extraction (R(2) = 0.85; n = 12; p < 0.0001). The trend toward increased Pb bioaccessibility in the elevated and anomalous subpopulations (75% ± 18% and 81% ± 8%, respectively) compared to background (63% ± 18%) is explained by the higher proportion of bioaccessible compounds used as pigments in older paints (Pb carbonate and Pb hydroxyl carbonate). This population-based study provides a nationally representative urban baseline for applications in human health risk assessment and risk management.

A novel class of macrocyclic peptidomimetics was identified and optimized as potent antagonists to the human motilin receptor (hMOT-R). Well-defined structure-activity relationships allowed for rapid optimization of potency that eventually led to high affinity antagonists to hMOT-R. Potency and antagonist functional activity were confirmed both in functional and cell-based assays, as well as on isolated rabbit intestinal smooth muscle strips. Rapid access to this novel class of macrocyclic target structures was made possible through two efficient and complementary solid-phase parallel synthetic approaches, both of which are reported herein.

More than 80 years of silver mining in the Cobalt area (Ontario, Canada) has led to widespread contamination of water with arsenic. The objective of this study was to determine the impact of changes in redox conditions on the stability of As in samples collected from a tailings wetland in the historic mining camp. Dissolved metal concentrations were monitored while tailings samples (approximately 1300 mg of As kg(-1), pH 7.4) were subjected to 30 days of reduction. Reoxidation of the samples was accomplished by air drying. The As oxidation states in the original, reduced, and reoxidized samples were determined using X-ray absorption spectroscopy (XAS). Arsenic speciation was affected by changes in redox conditions, resulting in rapid mobilization of As during reduction. Glucose input had a significant impact on the dissolution and speciation of As, suggesting that the As transformation was microbially mediated. When carbon was not limiting, the combination of reducing conditions and lower pH favored the formation of As(-I) species.

The long-term impacts of chronic metal exposure for aquatic biota are not well understood, partly due to a lack of understanding of metal speciation within tissues. The objective of this study was to determine hepatic Zn speciation of rainbow trout (Oncorhnychus mykiss) exposed to Zn-enriched water in relation to unexposed (control) fish,through direct analysis of freeze-dried liver samples using synchrotron X-ray absorption spectroscopy (XAS). Juvenile rainbow trout (n=30) were exposed to Zn in a two-step process, 200 microg L(-1) for 14 days, followed by 370 microg L(-1) for 23 days. Thirty other trout were grown in a control treatment (10 microg Zn L(-1)). At the end of the experiment, three liver samples per treatment were collected, freeze-dried, ground, and mixed homogeneously. Although Zn concentration was higher in the Zn-exposed livers than in the control livers (22.32 vs 13.73 mg kg(-1), respectively; p < 0.05), Zn speciation was similar for both groups. Extended X-ray absorption fine structure (EXAFS) spectroscopy indicated that Zn was coordinated to 4 sulfur atoms with an average Zn-S bond distance of 2.31 +/- 0.02 A. Sulfur K-XANES analysis confirmed that S was predominantly in reduced organic form analogous to cysteine. Our results are consistent with previous evidence for Zn(II) bonding to S in metallothionein proteins. These results suggest that the mechanisms for dealing with the extra load of bioaccumulated Zn in high exposure conditions were the same as in the control group.