Concentrations In Compartments Research Articles

0334 Use of salivary biomarkers to evaluate response to a stress management intervention

ObjectivesTo discuss methodological issues related to using salivary biomarkers to evaluate response to a stress management intervention.MethodFindings from a study which utilised salivary biomarkers to evaluate group responses to a...

Factors determining the accumulation of pentachlorophenol — a precursor of dioxins in bottom sediments of the Gulf of Gdańsk (Baltic Sea)

Abstract Pentachlorophenol (PCP) and its derivatives are considered to be the precursors of dioxins, thus their concentrations in environmental compartments remain relatively correlated. Unlimited production and usage of PCP in recent decades may have posed a potential ecological threat to marine ecosystems due to uncontrolled discharge of this contaminant into the Vistula River and finally into the Gulf of Gdańsk. Since there are no data on PCP concentration in sediments of the southern part of the Baltic Sea, the level of contamination has been examined and possible influence of sediment properties in the Gulf of Gdańsk on the accumulation intensification has been investigated. The study has resulted in the evaluation of an efficient analytical procedure characterized by a low detection limit (LOD<1 ng g−1 d.w.). Instrumental analyses have been supplemented with Microtox® bioassay in order to assess the sediment toxicity. The obtained concentrations in collected samples varied from below the LOD in sandy sediments to 179.31 ng g−1 d.w. in silty sediments, exceeding the PNEC value of 25 ng g−1 d.w. (Predicted No Effect Concentration) estimated for the Baltic Sea (Muir & Eduljee 1999). It has been proven that properties of sediments from the Gulf of Gdańsk, including pH, Eh of bottom water, the content of water and organic matter, affect the rate of PCP accumulation. High toxicity has been recorded in the bottom sediments of the Gdańsk Deep but no statistically significant correlation between PCP concentration and the sediment toxicity has been observed. Analysis of PCP concentration distribution in sediment cores revealed that the surface layer is the most polluted one, which indicates a continuous inflow of PCP from the Vistula River. Horizontal PCP distribution in the sediment from the Gdańsk Deep reveals variability similar to that observed for highly chlorinated dioxins (Niemirycz & Jankowska 2011).

Simplification of complex physiologically based pharmacokinetic models of monoclonal antibodies.

Monoclonal antibodies (mAbs) exhibit biexponential profiles in plasma that are commonly described with a standard two-compartment model with elimination from the central compartment. These models adequately describe mAb plasma PK. However, these models ignore elimination from the peripheral compartment. This may lead to underestimation of the volume of distribution of the peripheral compartment and thus over-predicts concentration in the peripheral compartment. We developed a simple and physiologically relevant model that incorporates information on binding and dissociation rates between mAb and FcRn receptor, mAb uptake, reflection, and catabolic degradation. We employed a previously published PBPK model and, with assumptions regarding rates of processes controlling mAb disposition, reduced the complex PBPK model to a simpler circular model with central, peripheral, and lymph compartments specifying elimination from both central and peripheral. We successfully applied the model to describe the PK of an investigational mAb. Our model presents an improvement over standard two-compartmental models in predicting whole-body average tissue concentrations while adequately describing plasma PK with minimal complexity and physiologically more meaningful parameters.

Pharmacokinetics and pharmacodynamics of propofol in children undergoing different types of surgeries

BackgroundPropofol is a commonly used agent in total intravenous anesthesia (TIVA). However, the link between its pharmacokinetics and pharmacodynamics has not been fully characterized in children yet. Our aim was to determine the quantitative relationship between the venous plasma concentration and bispectral index (BIS) effect in a heterogeneous group of pediatric patients undergoing various surgical procedures (ASA status I–III). MethodsNine male and nine female patients were anesthetized with propofol–fentanyl TIVA. Sparse venous samples for propofol concentrations assay and dense BIS measurements were collected during and after the end of infusion. Nonlinear mixed-effect modeling in NONMEM was used for data analysis. ResultsA three-compartment model was linked with a classical Emax model through a biophase compartment to describe the available data. All clearance and volume terms were allometrically scaled to account for the body mass difference among the patients under study. A typical patient had their PK parameters observed within the range of literature values for children. The pharmacodynamic parameters were highly variable. The EC50 of 2.80mg/L and the biophase distribution rate constant of 3.33min−1 were found for a typical patient. ConclusionsThe BIS values in children are highly correlated with the propofol effect compartment concentrations according to the classical Emax concentration–response relationship. Children had slightly lower sensitivity to propofol and slightly higher clearance, as compared with the adult data available in literature. The intra-patient variations in the BIS require the anesthesiologist's attention in using BIS values alone to evaluate the depth of anesthesia in children.

Experimental investigation of processing parameters and effects on chloralkali products in an electrolysis based chloralkali reactor

In the present study, the effects of different processing parameters on rate of hydrogen, chlorine and sodium hydroxide production are experimentally studied in a newly designed chloralkali process reactor. It is a multi membrane reactor which can also be coupled with a photochemical hydrogen production process. The parameters, which are studied in the present study, include applied voltage varied from 3V to 20V, brine concentration varied from 150g/L to 225g/L, electrolyte concentration in catholyte compartment varied from 35.3g/L to 58.8g/L, electrode height, temperature and different anode materials. A factorial design of experiments is applied and an analysis of variance (ANOVA) is used to analyze the experimental results. Energy and exergy efficiencies are also determined and discussed. The results show that graphite appears to be the most stable anode material. Brine and electrolyte concentration do not affect the production rate of products. Increasing the temperature increases the production but also increases the energy input to the system. Varying electrode height changes the current density which has a significant effect on rate of hydrogen production.

Regulation of RPE barrier function by all-trans retinoic acid in myopia

To investigate the effect of all-trans retinoic acid (atRA) on the barrier function of myopia retinal pigment epithelium (RPE), primary RPE cells from guinea pigs were cultured on the filters as monolayer and treated with 10−9M atRA. Epithelial permeability was measured by transepithelial electrical resistance (TER) and HRP concentration, western blot was used to observe the expression of intercellular junction-associated proteins. Treatment of primary cultures of confluent myopia RPE cells with atRA resulted in a progressive increase in TER and a progressive decrease of HRP concentration in the basolateral compartment. The expression of E-cadherin, Occludin and Claudin-1 proteins were up-regulated after treatment with atRA for 6h to 48h. We concluded that atRA could promote the epithelial barrier function of myopia RPE monolayer possibly by regulating expression of intercellular junction-associated proteins.

In Situ Sequestration of Hydrophobic Organic Contaminants in Sediments under Stagnant Contact with Activated Carbon. 2. Mass Transfer Modeling

The validity of a hydrophobic organic contaminant mass transfer model to predict the effectiveness of in situ activated carbon (AC) treatment under stagnant sediment-AC contact is studied for different contaminants and sediments. The modeling results and data from a previous 24-month column experiment of uptake in polyethylene samplers are within a factor of 2 for parent- and alkylated-polycyclic aromatic hydrocarbons in petroleum-impacted sediment and factors of 3-10 for polychlorinated biphenyls. The model successfully reproduces the relative effects of AC-sediment contact time, contaminant properties, AC particle size, AC mixing regime, AC distribution, and hydraulic conditions observed in the sediment column experiments. The model tracks contaminant concentrations in different sediment compartments over time, which provides useful information on the contaminant sequestration by the added AC. Long-term projection of the effectiveness of AC amendment using the model shows that the effects of AC particle size and particle-scale heterogeneity in AC distribution are pronounced within a year or so. However, the effect of those factors becomes less significant after a much longer contact period (on the order of a decade or two), resulting in substantial reduction in pore-water concentrations, for example, greater than 99% for benz[a]anthracene, under various scenarios.

Optimization of thermophilic anaerobic-aerobic treatment system for Palm Oil Mill Effluent (POME)

Optimization of an integrated anaerobic-aerobic bioreactor (IAAB) treatment system for the reduction of organic matter (Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD) and Total Suspended Solids (TSS) concentrations) in Palm Oil Mill Effluent (POME) to legal standards with high methane yield was performed for the first time under thermophilic condition (50°C–55°C) by using response surface methodology (RSM). The experiments were conducted based on a central composite rotatable design (CCRD) with three independent operating variables, organic loading rates in anaerobic compartment (OLRan) and mixed liquor volatile suspended solids (MLVSS) concentration in anaerobic (MLVSSan) and aerobic compartments (MLVSSa). The optimum conditions for the POME treatment were determined as OLRan of 15.6 g COD·L−1·d−1, MLVSSan of 43100 mg·L−1, and MLVSSa of 18600 mg·L−1, where high aerobic COD, BOD and TSS removal efficiencies of 96.3%, 97.9%, and 98.5% were achieved with treated BOD of 56 mg·L−1 and TSS of 28 mg·L−1 meeting the discharge standard. This optimization study successfully achieved a reduction of 42% in the BOD concentrations of the final treated effluent at a 48% higher OLRan as compared to the previous works. Besides, thermophilic IAAB system scores better feasibility and higher effectiveness as compared to the optimized mesophilic system. This is due to its higher ability to handle high OLR with higher overall treatment efficiencies (more than 99.6%), methane yield (0.31 L CH4·g−1 CODremoved) and purity of methane (67.5%). Hence, these advantages ascertain the applicability of thermophilic IAAB in the POME treatment or even in other high-strength wastewaters treatment.

Polymer capture by α-hemolysin pore upon salt concentration gradient

We have measured the rate of capture of single molecules of sodium poly(styrene sulfonate) by α-hemolysin protein pore by varying applied voltage, pH, and the salt concentration asymmetry across the pore. We show that electrostatic interaction between the polyelectrolyte and the protein pore significantly affects the polymer capture rate in addition to the enhancement of drift arising from electrolyte concentration gradient. At higher pH values where the electrostatic interaction between the polymer and the α-hemolysin pore is repulsive, an antagonistic coupling with the drift induced by salt concentration gradient emerges. This antagonistic coupling results in a nonmonotonic dependence of the polymer capture rate on the salt concentration in the donor compartment. The coupling between the pore-polymer interaction and drift can be weakened by increasing the strength of the electric field that drives the polymer translocation. In contrast, at lower pH values where the polymer-pore interaction is attractive, a synergy with the additional drift from salt concentration asymmetry arises and the capture rate depends monotonically on the donor salt concentration. For higher pH, we identify two regimes for the enhancement of capture rate by salt concentration gradient: (a) drift-dominated regime, where the capture rate is roughly quadratic in the ratio of salt concentration in the receiver compartment to that in the donor compartment, and (b) antagonistic coupling regime at higher values of this ratio with a linear relation for the polymer capture rate.

Chapter Two - Methods to Measure Intracellular Ca2 + Fluxes with Organelle-Targeted Aequorin-Based Probes

The photoprotein aequorin generates blue light upon binding of Ca(2+) ions. Together with its very low Ca(2+)-buffering capacity and the possibility to add specific targeting sequences, this property has rendered aequorin particularly suitable to monitor Ca(2+) concentrations in specific subcellular compartments. Recently, a new generation of genetically encoded Ca(2+) probes has been developed by fusing Ca(2+)-responsive elements with the green fluorescent protein (GFP). Aequorin has also been employed to this aim, resulting in an aequorin-GFP chimera with the Ca(2+) sensitivity of aequorin and the fluorescent properties of GFP. This setup has actually solved the major limitation of aequorin, for example, its poor ability to emit light, which rendered it inappropriate for the monitoring of Ca(2+) waves at the single-cell level by imaging. In spite of the numerous genetically encoded Ca(2+) indicators that are currently available, aequorin-based probes remain the method of election when an accurate quantification of Ca(2+) levels is required. Here, we describe currently available aequorin variants and their use for monitoring Ca(2+) waves in specific subcellular compartments. Among various applications, this method is relevant for the study of the alterations of Ca(2+) homeostasis that accompany oncogenesis, tumor progression, and response to therapy.

Chapter Three - Measuring Baseline Ca2 + Levels in Subcellular Compartments Using Genetically Engineered Fluorescent Indicators

Intracellular Ca(2+) signaling is involved in a series of physiological and pathological processes. In particular, an intimate crosstalk between bioenergetic metabolism and Ca(2+) homeostasis has been shown to determine cell fate in resting conditions as well as in response to stress. The endoplasmic reticulum and mitochondria represent key hubs of cellular metabolism and Ca(2+) signaling. However, it has been challenging to specifically detect highly localized Ca(2+) fluxes such as those bridging these two organelles. To circumvent this issue, various recombinant Ca(2+) indicators that can be targeted to specific subcellular compartments have been developed over the past two decades. While the use of these probes for measuring agonist-induced Ca(2+) signals in various organelles has been extensively described, the assessment of basal Ca(2+) concentrations within specific organelles is often disregarded, in spite of the fact that this parameter is vital for several metabolic functions, including the enzymatic activity of mitochondrial dehydrogenases of the Krebs cycle and protein folding in the endoplasmic reticulum. Here, we provide an overview on genetically engineered, organelle-targeted fluorescent Ca(2+) probes and outline their evolution. Moreover, we describe recently developed protocols to quantify baseline Ca(2+) concentrations in specific subcellular compartments. Among several applications, this method is suitable for assessing how changes in basal Ca(2+) levels affect the metabolic profile of cancer cells.

A Subspace-based Wiener System Identification Method for the Individualized Anesthesia Care

Abstract This study focuses on the individual sedation model identification problem, and proposes a subspace-based Wiener system identification method. The traditional compartmental pharmacokinetics-pharmacodynamics hypnosis model is considered as a specific Wiener system with the Hill equation nonlinear term. To deal with the Hill nonlinear term, the proposed method employs a set of specific bases to turn the Wiener system into a linear one, and then the subspace orthogonal projection identification method has been implemented to identify the transformed linear model. Compared with the traditional anesthesia model identification methods, the proposed method can effectively overcome the shortage of measurement data, get rid of the estimation of the effect compartment concentration which is impossible to be measured, and improve the individualization performance of the identified model. A simulation study on 24 various virtual patients from the Wang's Simulator has been conducted and validates the efficiency and robustness of the proposed method, and a drug infusion instruction has been provided in order to get relatively accurate identification performance.

Paracetamol: a focus for the general pediatrician

Paracetamol (acetaminophen) is one of the most popular and widely used drugs for the treatment of pain and fever in children. This drug has multiple mechanisms of action, but its pharmacodynamic is still not well known. The central nervous system is the main site of action and it mirrors the paracetamol effect compartment. The recommended dosages and routes of administration should be different whether paracetamol is used for the treatment of pain or fever. For example, the rectal route, while being efficacious for the treatment of fever, should be avoided in pain management. Paracetamol is a safe drug, but some clinical conditions and concomitant drugs, which are frequent in clinical practice, may increase the risk of paracetamol toxicity. Therefore, it is important to optimize its administration to avoid overdoses and maximize its effect. The principal mediator of the paracetamol toxicity is the N-acetyl-p-benzo-quinone imine (NAPQI), a toxic product of the paracetamol metabolism, which could bind cysteine groups on proteins forming paracetamol-protein adduct in the liver. Although frequently prescribed, the concept of "effect compartment concentration" and the possible co-factors that could cause toxicity at recommended doses are not familiar to all pediatricians and general practitioners. We reviewed the literature concerning paracetamol mechanisms of action, we highlighted some relevant pharmacodynamic concepts for clinical practice, and we summarized the possible risk factors for toxicity at therapeutic dosages.

Pharmacokinetic–pharmacodynamic evaluation of the major component astragaloside IV on the immunomodulatory effects of Yu-ping-feng prescription

Yu-ping-feng decoction (YPF), a traditional Chinese medicine (TCM) prescription, is widely used to treat some respiratory tract diseases. This study was aimed to set up a pharmacokinetic-pharmacodynamic (PK-PD) model to assess dose-effect relationships for immunomodulatory effects of YPF in rats and to clarify compatibility mechanisms for TCM prescription system. Serum samples taken after YPF administration were tested on spleen cells of rats in vitro and proliferation ratio of spleen cells was used as end points to evaluate pharmacodynamic properties of immunoregulatory effects of YPF prescription. And with a HPLC-MS method, concentrations of Astragaloside IV (AS), a main component of YPF, were determined to achieve pharmacokinetic parameters after administration of a simplified prescription which is composed with AS, Atractylenolide I and Prim-O-glucosylcimifugin which are representative components of YPF. A plot of serum AS concentrations versus time and effects showed that there was a correlated relationship between AS concentrations and effects of YPF, and the concentration-response curve which was based on an E max model showed a counterclockwise hysteresis manner. A PK-PD model with Sheiner's method was used to describe time course of AS concentration in blood compartment and effect compartment, and main parameters with the PK-PD model were calculated. These results showed that there is a symmetry relationship between serum AS concentrations and responses of serum containing medicine of YPF prescription, which means that AS plays an important role in immunoregulatory effects of YPF. And the investigation on dose-effect relationships has displayed a feasible method to clarify mechanisms of combination for TCM prescriptions.

Chemerin concentrations in maternal and fetal compartments: implications for metabolic adaptations to normal human pregnancy

Chemerin, a novel adipocytokine, has been implicated in major metabolic and inflammatory processes. Study aims were to determine whether circulating maternal chemerin concentration (1) differs between pregnant and non-pregnant women, (2) changes as a function of gestational age, and (3) correlates with maternal insulin resistance. In addition, we investigated which compartment, maternal, fetal or placental, is the source of chemerin in maternal circulation. The study included three groups: Non-pregnant (n=18), pregnant women in the first trimester (n=19) and pregnant women in the third trimester (n=33). Chemerin was measured in cord blood and in maternal serum samples taken before and after delivery. Chemerin mRNA expression was evaluated in fetal and human adult tissues. Chemerin serum concentration was significantly higher in pregnant women in the third trimester than in non-pregnant and pregnant women in the first trimester. Chemerin concentration positively correlated with body mass index (BMI) and insulin resistance. Antenatal chemerin concentration was significantly lower than that during the postpartum period. Neonatal chemerin did not correlate with maternal one. Chemerin mRNA expression was abundant in fetal and adult liver and omental fat, but relatively low in placenta. Chemerin is increased during normal gestation and is associated with maternal BMI and insulin resistance. Maternal tissues, possibly liver and adipose tissue, contribute to the increased maternal chemerin concentration.

Utilization of an ex vivo human placental perfusion model to predict potential fetal exposure to carboplatin during pregnancy

The objective of this study was to determine the fetal drug compartment concentrations when various concentrations of carboplatin cross the placental-trophoblastic barrier and the effect on the fetal kidneys. An ex vivo human placenta perfusion model was utilized. Term human placentae (n = 9) were collected immediately after delivery and then reperfused with plasma concentrations achieved with carboplatin an area under the curve of 5 (1000 ng/mL), 7.5 (5000 ng/mL), or 11 (11,000 ng/mL). Antipyrine was used as a reference compound. Samples were collected over 2 hours. Placental transfer was evaluated by computation of transport fraction and clearance index. Primary cells isolated by explant culture of 16-18 week old fetal organ tissues were incubated with carboplatin for up to 48 hours with untreated cell as controls. Immunohistochemical, flow cytometry analysis, and immunoblotting were applied for the expression of apoptosis-related proteins. Mean transport fractions for carboplatin at low, middle, and high concentrations were 0.05 ± 0.02, 0.04 ± 0.01, and 0.10 ± 0.01, respectively, with clearance indexes of 0.22 ± 0.01, 0.14 ± 0.08, and 0.50 ± 0.07, respectively. The fetal peak concentrations of carboplatin achieved were 61 ± 39 ng/mL (low), 375 ± 248 ng/mL (middle), and 2081 ± 529 ng/mL (high). Fetal kidney cells exposed to carboplatin showed a concentration-dependent increased expression of apoptosis-inducing factor and p53 apoptosis proteins and a time-dependent increase in expression Bax apoptosis protein expression. Apoptosis was confirmed at the high concentration by flow cytometry. Doses of carboplatin up to an area under the curve of 7.5 were not associated with significant placental transfer, fetal exposure, or fetal toxic effects. This suggests it might not be necessary to empirically reduce carboplatin doses in pregnant women.

Secretory expression of biologically active human Herpes virus interleukin-10 analogues in Escherichia colivia a modified Sec-dependent transporter construct

BackgroundInterleukin-10 homologues encoded by Herpes viruses such as Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) hold interesting structural and biological characteristics compared to human interleukin-10 (hIL-10) that render these proteins promising candidates for therapeutic application in inflammatory bowel disease (IBD). Intestinal delivery of cytokines using bacterial carriers as chassis represents a novel approach for treatment of IBD patients. For proof of concept, a Sec-dependent transporter construct was designed for secretory expression of recombinant viral IL-10 proteins in the periplasm of Escherichia coli laboratory strain BL21 (DE3), which might serve as part of a prospective lysis based delivery and containment system.ResultsThe signal peptide of E. coli outer membrane protein F fused to the mature form of the viral IL-10 proteins enabled successful transport into the periplasm, a compartment which seems crucial for proper assembly of the dimeric configuration of the cytokines. Cytokine concentrations in different bacterial compartments were determined by ELISA and achieved yields of 67.8 ng/ml ± 24.9 ng/ml for HCMV IL-10 and 1.5 μg/ml ± 841.4 ng/ml for EBV IL-10 in the periplasm. Immunoblot analysis was used to confirm the correct size of the E. coli-derived recombinant cytokines. Phosphorylation of signal transducer and activator of transcription 3 (STAT3) as part of the signal transduction cascade after IL-10 receptor interaction, as well as suppression of tumor necrosis factor α (TNF-α) release of lipopolysaccharide-stimulated mouse macrophages were used as read-out assays for proving in vitro biological activity of the E. coli derived, recombinant viral IL-10 counterparts.ConclusionsIn this study, proof of principle is provided that E. coli cells are a suitable chassis for secretory expression of viral IL-10 cytokines encoded by codon-optimized synthetic genes fused to the E. coli ompF signal sequence. In vitro biological activity evidenced by activation of transcription factor STAT3 and suppression of TNF-α in mammalian cell lines was shown to be strictly dependent on export of viral IL-10 proteins into the periplasmic compartment. E. coli might serve as carrier system for in situ delivery of therapeutic molecules in the gut, thus representing a further step in the development of novel approaches for treatment of IBD.

Determination of gas diffusion coefficients in saturated porous media: He and CH4 diffusion in Boom Clay

Boom Clay is presently studied as the reference host formation for the disposal of high-level and long-lived radioactive waste in Belgium. In a geological repository, the production of gas is unavoidable. Gas is produced by different mechanisms: anaerobic corrosion of metals in waste and packaging, radiolysis of water and organic materials in the packages and microbial degradation of various organic wastes. Corrosion and radiolysis yield mainly hydrogen while microbial degradation leads to methane and carbon dioxide. The generated gas will dissolve in the ground water. As transport in Boom Clay is dominated by diffusion, the dissolved gas is transported away from the repository by diffusion as dissolved species. If the rate of gas generation is larger than the diffusive flux into Boom Clay, the pore water within the disposal gallery will become oversaturated and a free gas phase might form. If the gas pressure keeps increasing, free gas ingress into Boom Clay will occur, most likely through creation of new pathways. In order to make a good evaluation of the balance between gas generation and gas dissipation through engineered barriers and host formation, good estimates for the gas diffusion coefficients of the gases are needed. The currently available gas diffusion parameters for hydrogen in Boom Clay suffer from a large uncertainty, and by application of conservative values for both the source term and the gas migration term the formation of a free gas phase can presently not be excluded for some waste types.In this study a versatile method was developed to determine more precisely the gas diffusion coefficient for dissolved gases in Boom Clay. For the development of the technique, He and CH4 were used.The proposed method is based on a through diffusion methodology and allows for two dissolved gases to diffuse through a clay sample at the same time. From the evolution of the diffusant concentration in both compartments, the apparent diffusion coefficients of dissolved He and CH4 were obtained: 12.2×10−10 and 2.42×10−10m2/s, with uncertainties of 10%, respectively.

Hydrograph separation to improve understanding of Dissolved Organic Carbon Dynamics in Headwater catchments

AbstractDissolved organic carbon (DOC) is a key component of the global carbon cycle, but, to date, large uncertainties still exist on its source and fate in first‐order streams. In a 23 ha rangeland and steep‐slope headwater of South Africa, our aim was to quantify the contribution of overland flow (OF), soil water (SW) and ground water (GW) to DOC fluxes (DOCF), and to interpret the results in terms of DOC sources and fate. The average 2010–2011 DOC concentration (DOCC) at the catchment outlet was 4.7 mg C l−1 with a standard error of ±2.5 mg C l−1, which was significantly lower than in SW (15.2 ± 1.6 mg C l−1) and OF (11.9 ± 0.8 mg C l−1), but higher than in GW (2.3 ± 0.6 mg C l−1). Based on end‐member mixing using Si and Na concentration in the water compartments, the average SW contribution to DOCF was 66.4%, followed by OF (30.0%) and GW (3.6%). The resulting estimated DOCF at the catchment outlet was 8.05 g C m2 y−1. This was much higher than the observed value of 2.80 g C m2 y−1, meaning that 5.25 g C m2 y−1 or 65% of the DOC is lost during its downslope and/or downstream transport to the catchment outlet. Complementary investigations revealed that the DOCC in SW dropped from 15.2 ± 1.6 to 2.6 ± 0.3 mg C l−1 during its downslope transport to the river system, which corresponded to a net loss of 5.10 g C m2 y−1, or 97% of the catchment DOC losses. These results on DOC sources and potential fate in headwaters are expected to improve our understanding of the impact of hydrology on the global C‐cycle. Copyright © 2013 John Wiley & Sons, Ltd.

The self‐association of the cyclotide kalata B2 in solution is guided by hydrophobic interactions

The cyclotides are a family of small head-to-tail cyclic plant defense proteins. In addition to their cyclic backbone, cyclotides comprise three disulfide bonds in a knotted arrangement, resulting in a highly cross-braced structure that provides exceptional chemical and proteolytic stability. A number of bioactivities have been associated with cyclotides, including insecticidal, antimicrobial, anti-viral and cytotoxic, and these activities are related to an ability to target and disrupt biological membranes. Kalata B2 and to a lesser extent kalata B1, isolated from Oldenlandia affinis, self-associate to tetramers and octamers in aqueous buffers, and this oligomerization has been suggested to be relevant for their ability to form pores in membranes. Here we demonstrate by solution NMR spectroscopy analysis that the oligomerization of kalata B2 is concentration dependent and that it involves the packing of hydrophobic residues normally exposed on the surface of kalata B2 into a multimeric hydrophobic core. Interestingly, the hydrophobic surface that is "quenched" has previously been shown to be responsible for the ability of kalata B2 to insert into membranes. Thus, it seems unlikely that the oligomers observed in aqueous solution are related to any multimeric state present in a membrane environment, and responsible for the formation of pores. The ability to self-associate might alternatively provide a mechanism for preventing self-toxicity when stored at high concentrations in intracellular compartments.