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Study region: Bavaria, Germany. Study focus: The Altmühl River is prone to nutrient inputs from agricultural activities. Quantifying nitrate nitrogen (NO3−-N) and total phosphorus (TP) concentrations due to potential future changes in the watershed is necessary for managing water quality and adhering to water policy directives. The Soil and Water Assessment Tool (SWAT) was used to provide stakeholders with support in determining the impacts of climate change (CC) in combination with crop land use change (LUC) scenarios on streamflow, NO3−-N and TP to the 2050 time horizon. The CC simulations stemmed from RCMs and the LUC scenarios were developed with stakeholders. New hydrological insights for the region: When CC was combined with LUC, mean annual NO3−-N loads increased 3-fold, and TP loads 8-fold, compared to the CC simulations alone. Nutrient loads were higher in several months due to the future increased annual precipitation plus the additional fertilizer input in the land use scenarios. The maize areas above the Altmühl Lake contributed greatly to TP loads, while winter wheat areas mainly contributed to NO3−-N loads. When CC was combined with LUC, the in-stream nutrient concentrations exceeded ministerial guidelines of 11 mg TP/L and 0.05 mgNO3−-N/L every month at the outlet. CC simulations combined with LUC scenarios demonstrated non-linear dynamics whereby the direction and the magnitude of impacts were not predictable from the individual changes alone.

There are a wide range of benign and malignant pathologies that the radiologist may encounter in the adrenal glands and kidneys, often incidentally when imaging is performed for other indications. Many imaging modalities including CT, MR, and US are often used in an attempt to characterize these lesions. A definitive radiological diagnosis, however, is not always possible. This is at times due to atypical presentations of typical lesions which may be mistaken for more aggressive or concerning pathologic conditions. Adrenal lesions that do not demonstrate characteristic benign imaging features might require surgical excision. Similarly, cystic renal lesions that demonstrate nodular enhancement are concerning for Bosniak IV lesions and require surgical management. We report 3 cases in 3 different patients of incidentally discovered hematomas with peripheral enhancement, 2 involving the adrenal gland and 1 involving the kidney. All 3 of these histologically proven hematomas demonstrated similar radiological manifestations of peripheral nodular progressive enhancement, mimicking neoplastic conditions, and necessitating surgical removal.

Purpose: Yttrium-90 (90Y) radioembolization with an escalated dose has been shown to improve clinical outcomes compared with standard dose radioembolization, but there are few data on the local control of primary liver tumors. We reported the clinical outcomes of patients with unresectable primary liver tumors treated with 90Y radioembolization with an escalated dose. Methods and Materials: Clinical data of patients with unresectable hepatocellular carcinoma (HCC), cholangiocarcinoma (CC), and biphenotypic tumors (cHCC-CC) treated with radioembolization with an escalated dose (≥150 Gy) between 2013 and 2020 with >3 months follow-up were retrospectively reviewed. The primary endpoint was freedom from local progression. Clinical response was defined by Modified Response Evaluation Criteria in Solid Tumours and toxic effects were assessed using Common Terminology Criteria for Adverse Events version 5.0. Results: Fifty-three patients with HCC and 15 patients with CC/cHCC-CC were analyzed. The median dose delivered was 205 Gy (interquartile range, 183-253 Gy) and 198 Gy (interquartile range, 154-234 Gy) for patients with HCC and CC/cHCC-CC, respectively. The 1-year freedom from local progression rate was 54% (95% confidence interval [CI], 38%-78%) for patients with HCC and 66% (95% CI, 42%-100%) for patients with CC/cHCC-CC. For patients with HCC, United Network for Organ Sharing nodal stage 1 (P = .01), nonsolitary tumors (P = .02), pretreatment α-fetoprotein of >7.7 ng/mL (P = .006), and ≤268 Gy dose delivered (P = .003) were predictors for local progression on multivariate Cox analysis. No patients with HCC who received a dose >268 Gy had a local tumor progression. The 1-year overall survival for patients with HCC was 74% (95% CI, 61%-89%). After radioembolization, 5 (7%) patients had grade 3 ascites, and 4 (6%) patients had grade 3/4 hyperbilirubinemia. Conclusions: Treatment of unresectable primary liver tumors with 90Y radioembolization with an escalated dose was safe and well tolerated. Delivery of >268 Gy may improve local tumor control of HCC. Determination of the maximum tolerated dose needs to be performed in the context of future prospective dose-escalation trials to further evaluate the safety and efficacy of such an approach.

We investigated the bioelectrochemical properties of an FAD-dependent glucose dehydrogenase from Trichoderma virens (TvGDH) and its electrochemical behaviour when immobilized on a graphite electrode. TvGDH was recently shown to have an unusual substrate spectrum and to prefer maltose over glucose as substrate, and hence could be of interest as recognition element in a maltose sensor. In this study, we determined the redox potential of TvGDH, which is -0.268 ± 0.007 V vs. SHE, and advantageously low to be used with many redox mediators or redox polymers. The enzyme was entrapped in, and wired by an osmium redox polymer (poly(1-vinylimidazole-co-allylamine)-{[Os(2,2'-bipyridine) 2 Cl]Cl}) with formal redox potential of +0.275 V vs. Ag|AgCl via poly(ethylene glycol) diglycidyl ether crosslinking onto a graphite electrode. When the TvGDH-based biosensor was tested with maltose it showed a sensitivity of 1.7 μA mM -1 cm -2 , a linear range of 0.5-15 mM, and a detection limit of 0.45 mM. Furthermore, it gave the lowest apparent Michaelis-Menten constant (K M app ) of 19.2 ± 1.5 mM towards maltose when compared to other sugars. The biosensor is also able to detect other saccharides including glucose, maltotriose and galactose, these however also interfere with maltose sensing., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Managing blood glucose can affect important clinical outcomes during the intraoperative phase of surgery. However, currently available instruments for glucose monitoring during surgery are few and not optimized for the specific application. Here we report an attempt to exploit an enzymatic sensor in a vein replica that could continuously monitor glucose level in an authentic human bloodstream. First, detailed investigations of the superficial venous systems of volunteers were carried out using ocular and palpating examinations, as well as advanced ultrasound measurements. Second, a tubular glucose-sensitive biosensor mimicking a venous system was designed and tested. Almost ideal linear dependence of current output on glucose concentration in phosphate buffer saline was obtained in the range 2.2-22.0 mM, whereas the dependence in human plasma was less linear. Finally, the developed biosensor was investigated in whole blood under homeostatic conditions. A specific correlation was found between the current output and glucose concentration at the initial stage of the biodevice operation. However, with time, blood coagulation during measurements negatively affected the performance of the biodevice. When the experimental results were remodeled to predict the response without the influence of blood coagulation, the sensor output closely followed the blood glucose level., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Phenotypic plasticity drives cancer progression, impacts treatment response, and is a major driver of therapeutic resistance. In melanoma, a regulatory axis between the MITF and BRN2 transcription factors has been reported to promote tumor heterogeneity by mediating switching between proliferative and invasive phenotypes, respectively. Despite strong evidence that subpopulations of cells that exhibit a BRN2 high /MITF low expression profile switch to a predominantly invasive phenotype, the mechanisms by which this switch is propagated and promotes invasion remain poorly defined. We have found that a reciprocal relationship between BRN2 and NOTCH1/2 signaling exists in melanoma cells in vitro, within patient datasets, and in in vivo primary and metastatic human tumors that bolsters acquisition of invasiveness. Working through the epigenetic modulator EZH2, the BRN2‒NOTCH1/2 axis is potentially a key mechanism by which the invasive phenotype is maintained. Given the emergence of agents targeting both EZH2 and NOTCH, understanding the mechanism through which BRN2 promotes heterogeneity may provide crucial biomarkers to predict treatment response to prevent metastasis., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

To efficiently decompose biomass, fungi have developed various enzymatic and non-enzymatic strategies and are a source of versatile biocatalysts. The endoglucanases in glycosyl hydrolase CAZy family 45 (GH45) are known for their small size, a high thermostability and a broad substrate specificity that has been employed in textile and detergent industries. Here we report the heterologous expression and characterisation of an GH45 endoglucanase from the brown rot Fomitopsis pinicola and its direct comparison to an already characterised GH45 from the white rot Phanerochaete chrysosporium. Both enzymes were recombinantly expressed in Pichia pastoris and purified by two chromatographic steps. The biochemical characterisation highlighted the acidophilic character, with an optimal pH of 4, and a preference for amorphous substrates as carboxymethyl cellulose (CMC) and substrates containing β-1,4-glucans rather than the previously reported β-1,3/1,4-glucans lichenan and β-glucan. The dominating products from β-1,4-glucans were C3-C6 oligosaccharides, whereas from β-1,3/1,4-glucans glucose was the main reaction product. From the characterisation no differences between the brown rot and the white rot GH45 was evident., (Copyright © 2022. Published by Elsevier Inc.)

The cellular mechanisms required to ensure homeostasis of the hematopoietic niche and the ability of this niche to support hematopoiesis upon stress remain elusive. We here identify Wnt5a in Osterix+ mesenchymal progenitor and stem cells (MSPCs) as a critical factor for niche-dependent hematopoiesis. Mice lacking Wnt5a in MSPCs suffer from stress-related bone marrow (BM) failure and increased mortality. Niche cells devoid of Wnt5a show defective actin stress fiber orientation due to an elevated activity of the small GTPase CDC42. This results in incorrect positioning of autophagosomes and lysosomes, thus reducing autophagy and increasing oxidative stress. In MSPCs from patients from BM failure states which share features of peripheral cytopenia and hypocellular BM, we find similar defects in actin stress fiber orientation, reduced and incorrect colocalization of autophagosomes and lysosomes, and CDC42 activation. Strikingly, a short pharmacological intervention to attenuate elevated CDC42 activation in vivo in mice prevents defective actin-anchored autophagy in MSPCs, salvages hematopoiesis and protects against lethal cytopenia upon stress. In summary, our study identifies Wnt5a as a restriction factor for niche homeostasis by affecting CDC42-regulated actin stress-fiber orientation and autophagy upon stress. Our data further imply a critical role for autophagy in MSPCs for adequate support of hematopoiesis by the niche upon stress and in human diseases characterized by peripheral cytopenias and hypocellular BM., (© 2022 by The American Society of Hematology.)

Enzymatic biofuel cells utilize oxidoreductases as highly specific and highly active electrocatalysts to convert a fuel and an oxidant even in complex biological matrices like hydrolysates or physiological fluids into electric energy. The hemoflavoenzyme cellobiose dehydrogenase is investigated as a versatile bioelectrocatalyst for the anode reaction of biofuel cells, because it is robust, converts a range of different carbohydrates, and can transfer electrons to the anode by direct electron transfer or via redox mediators. The versatility of cellobiose dehydrogenase has led to the development of various electrode modifications to create biofuel cells and biosupercapacitors that are capable to power small electronic devices like biosensors and connect them wireless to a receiver., (Copyright © 2021. Published by Elsevier Ltd.)

Glyoxal oxidase (GLOX) is an extracellular source of H 2 O 2 in white-rot secretomes, where it acts in concert with peroxidases to degrade lignin. It has been reported that GLOX requires activation prior to catalytic turnover and that a peroxidase system can fulfill this task. In this study, we verify that an oxidation product of horseradish peroxidase, the radical cation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), is an activator for GLOX. A spectroelectrochemical cell was used to generate the activating radical species, to continuously measure its concentration, and to simultaneously measure the catalytic activity of GLOX based on its O 2 consumption. The results show that GLOX can undergo multiple catalytic turnovers upon activation and that activity increases with the activator concentration. However, we also found that the ABTS cation radical can serve as an electron acceptor which becomes visible in the absence of O 2 . Furthermore, GLOX activity is highly restrained by the naturally occurring, low O 2 concentration. We conclude that GLOX is indeed an auxiliary enzyme for H 2 O 2 production in white-rot secretomes. Its turnover rate is strongly regulated by the availability of O 2 and the radical generating activity of peroxidases present in the secretome, which acts as a feedback loop for GLOX activity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Increasing temperatures and snow scarcity pose a serious threat to ski tourism. While the impacts of climate change on ski tourism have been elaborated extensively, little is known so far on the vulnerability of winter tourism towards both internal climate variability and climate change. We use a 50-member single model large ensemble from a regional climate model to drive the physically-based snowpack model SNOWPACK for eight stations across the Swiss Alps to model daily snow depth, incorporating both natural snow conditions and including technical snow production. We make a probabilistic assessment of the vulnerability of ski tourism towards internal climate variability in a future climate by analyzing selected tourism-related snow indicators and find significant overall decrease in snow reliability in the future. Further, we show how the sensitivity towards internal climate variability differs among different tourism-related snow indicators and find that certain indicators are more vulnerable to internal climate variability than others. We show that technical snow production is an appropriate adaptation strategy to tackle risks from climate change and internal climate variability. While technical snow production can drastically reduce uncertainties related to internal climate variability, in low elevations, the technique reaches its limits to counteract global warming by the mid of the century., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Laccase is predominantly found in lignin degrading filamentous white rot fungi, where it is involved in the oxidative degradation of this recalcitrant heteropolymer. In brown rot fungi it is much less prevalent: laccases from only a few brown rots have been detected and only two have been characterized. This study tries to understand the role of this ligninolytic enzyme in brown rots by investigating the catalytic properties of laccases secreted by Fomitopsis pinicola FP58527 SS1. When grown on either poplar or spruce wood blocks, several laccases were detected in the secretome. Two of them (FpLcc1 and FpLcc2) were heterologously produced using Trichoderma reesei QM9414 Δxyr1 as expression host and purified to homogeneity by consecutive steps of hydrophobic interaction, anion exchange and size exclusion chromatography. With the substrates 2,2-azino-bis(3-ethylthiazoline-6-sulfonate) (ABTS), 2,6-dimethoxyphenol (2,6-DMP) and guaiacol both laccases showed similar, low pH-optima below 3 for ABTS and 2,6-DMP and at pH 3.5 for guaiacol which is at the acidic end of laccases isolated from white rot fungi. The determined K M values were low while k cat values measured at acidic conditions were comparable to those reported for other laccases from white rot fungi. While both enzymes showed a moderate decrease in activity in the presence of oxalic and citric acid FpLcc2 was activated by acetic acid up to 3.7 times. This activation effect is much more pronounced at pH 5.0 compared to pH 3.0 and could already be observed at a concentration of 1 mM acetic acid., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

In the secretome of Phanerochaete chrysosporium, a white-rot fungus serving as a model organism to elucidate lignocellulose deconstruction, the copper containing metalloprotein glyoxal oxidase (GLOX) is potentially involved in the crucial production of hydrogen peroxide to fuel and initiate oxidative biomass degradation by lignin-degrading peroxidases. Its ability to oxidize a variety of aldehydes and α-hydroxy carbonyls with the concomitant reduction of dioxygen to hydrogen peroxide has attracted attention for its application as green biocatalyst in different industrial fields. Here we report and compare two efficient processes for the heterologous production of GLOX from P. chrysosporium using the well-established methanolytic yeast Pichia pastoris and the filamentous fungus Trichoderma reesei as expression hosts with subsequent purification by anion exchange and hydrophobic interaction chromatography. Both processes were shown to be suitable for the production of the target protein at high levels. GLOX produced in T. reesei carries mainly Man5 glycosylation while the enzyme produced in P. pastoris exhibits the typical high-mannose type N-glycosylation. The enzyme expressed in P. pastoris showed slightly higher specific activities which correlates with the higher copper loading of 65.5 % compared to 51.9 % for the protein from T. reesei. The pH optimum for both recombinant proteins was 6.0, however, GLOX activity was found to be highly affected by different buffer species. Both enzymes showed very similar substrate affinities and turnover numbers with the highest catalytic efficiency observed for methylglyoxal. GLOX from both expression hosts is therefore a suitable enzyme for further mechanistic characterization and application studies., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Urbanization, agriculture, and the manipulation of the hydrological cycle are the main drivers of multiple stressors affecting river ecosystems across the world. Physical, chemical, and biological stressors follow characteristic patterns of occurrence, intensity, and frequency, linked to human pressure and socio-economic settings. The societal perception of stressor effects changes when moving from broad geographic regions to narrower basin or waterbody scales, as political and ecologically based perspectives change across scales. Current approaches relating the stressor effects on river networks and human societies fail to incorporate complexities associated to their co-occurrence, such as: i) the evidence that drivers can be associated to different stressors; ii) their intensity and frequency may differ across spatial and temporal scales; iii) their differential effects on biophysical receptors may be related to their order of occurrence; iv) current and legacy stressors may produce unexpected outcomes; v) the potentially different response of different biological variables to stressor combinations; vi) the conflicting effects of multiple stressors on ecosystem services; and, vii) management of stressor effects should consider multiple occurrence scales. We discuss how to incorporate these aspects to present frameworks considering biophysical and societal consequences of multiple stressors, to better understand and manage the effects being caused on river networks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: We investigated whether residing in places with higher greenness, more trees and more allergenic trees early in life increases the risk of allergic outcomes, and whether these associations differ depending on the concentration of air pollutants., Methods: The analytic sample included 631 children from the German birth cohort LISA Leipzig. Asthma and allergic rhinitis, sensitization to aeroallergens and food allergens, as well as confounders, were collected prospectively up to 15 years. Greenness was assessed by Normalized Difference Vegetation Index (NDVI). A tree registry was used to derive information on trees, which were classified into allergenic and non-allergenic. Annual average concentrations of nitrogen dioxide (NO 2 ) and ozone were also used. Geographic exposures were assigned to home addresses at birth. Longitudinal associations were analysed using generalized estimating equations., Results: Medium and high numbers (tertiles) of trees and allergenic trees in a 500 m buffer around birth addresses were associated with increased odds of allergic rhinitis up to 15 years regardless of NDVI. These exposures were also related to higher odds of sensitization to aeroallergens. Associations with asthma and sensitization to food allergens were less consistent. Effect estimates for allergic rhinitis were stronger in the high tertile of NO 2 compared to the low tertile, while an opposite tendency was observed for ozone., Conclusion: We observed that early life residence in places with many trees, and allergenic trees specifically, may increase the prevalence of allergic rhinitis later in life. This association and its modification by air pollution should be pursued in further studies., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Lifestyle interventions to increase exercise and improve diet have been the focus of recent clinical trials to potentially prevent Alzheimer's disease (AD). However, despite the strong links between sleep disruptions, cognitive decline, and AD, sleep enhancement has yet to be targeted as a lifestyle intervention to prevent AD. A recent meta-analysis suggests that approximately 15% of AD may be prevented by an efficacious intervention aimed to reduce sleep disturbances and sleep disorders. Chronic insomnia is the most frequent sleep disorder occurring in at least 40% of older adults. Individuals with insomnia are more likely to be diagnosed with Alzheimer's Disease (AD) and demonstrate decline in cognitive function at long-term follow-up. AD is characterized by the accumulation of amyloid-β (Aβ) plaques and tau tangles in the brain, and growing evidence shows impaired sleep contributes to the accumulation of Aβ. An intervention aimed at improving insomnia may be a critical opportunity for primary prevention to slow cognitive decline and potentially delay the onset of AD. Cognitive behavioral therapy for insomnia (CBT-I) is an efficacious treatment for insomnia, but the use of CBT-I to improve cognitive function and potentially reduce the rate of Aβ accumulation has never been examined. Therefore, the objective of the proposed study is to examine the efficacy of CBT-I on improving cognitive function in older adults with symptoms of insomnia. An exploratory aim is to assess the effect of CBT-I on rate of Aβ accumulation., (Copyright © 2020. Published by Elsevier Inc.)

Epidermolysis bullosa acquisita is an autoimmune skin disease characterized by subepidermal blisters. The pathogenesis is mediated by deposits of autoantibodies directed against type VII collagen in the skin, but the sequence of events regulating the localization of skin blisters is not fully understood. In this study, using the immunization-induced mouse model of epidermolysis bullosa acquisita, we demonstrate that epidermal disruption induces not only an infiltration of CD4 + T cells but also a T helper type 1 phenotype as it has been described for delayed-type hypersensitivity reactions. This T helper type 1 reaction was not found when different antigens were applied. Deep T-cell receptor β profiling revealed shifts in the V/J gene usage only in epidermolysis bullosa acquisita, suggesting an infiltration of autoantigen-specific T cells. To target these autoantigen-specific T cells, we established an approach with which skin inflammation could be prevented without impairing the functionality of autoantibodies. We conclude that T-cell involvement in skin blistering diseases such as epidermolysis bullosa acquisita relates not only to T-cell help for B cells that produce pathogenic autoantibodies but also to autoreactive T helper type 1 effector cells that migrate into injured skin sites, exacerbate inflammation through production of inflammatory cytokines such as IFNγ, and prevent wound healing., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Polymicrobial biofilm related infections have been a major threat in health care. In this study, the co-immobilization of cellobiose dehydrogenase (CDH) and deoxyribonuclease I (DNase) on positively charged chitosan nanoparticles (CSNPs) resulted in a bi-functional nanoparticle (CSNP-DNase-CDH) targeting both biofilm matrix and microorganisms. The in-vitro antibiofilm activities of CSNPs against monomicrobial and polymicrobial biofilms of Candida albicans and Staphylococcus aureus were evaluated. The results showed that CSNPs were able to penetrate across the matrix of biofilms and interfere with embedded microbial cells. CSNP-DNase-CDH exhibited a higher activity than CSNPs loaded with only DNase or CDH for inhibiting monomicrobial and polymicrobial biofilm formation as well as for disrupting pre-formed biofilms. Furthermore, CSNP-DNase-CDH could disrupt the biofilm formation through degradation of eDNA, reduce biofilm thickness, and kill microbial cells on silicone. The bi-functional CSNP is applicable for the protection of medical devices from polymicrobial biofilms or the treatment of device associated infections., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Pyranose oxidase (POx) catalyzes the oxidation of d-glucose to 2-ketoglucose with concurrent reduction of oxygen to H 2 O 2 . POx from Trametes ochracea (ToPOx) is known to react with alternative electron acceptors including 1,4-benzoquinone (1,4-BQ), 2,6-dichlorophenol indophenol (DCPIP), and the ferrocenium ion. In this study, enzyme variants with improved electron acceptor turnover and reduced oxygen turnover were characterized as potential anode biocatalysts. Pre-steady-state kinetics of the oxidative half-reaction of ToPOx variants T166R, Q448H, L545C, and L547R with these alternative electron acceptors were evaluated using stopped-flow spectrophotometry. Higher kinetic constants were observed as compared to the wild-type ToPOx for some of the variants. Subsequently, the variants were immobilized on glassy carbon electrodes. Cyclic voltammetry measurements were performed to measure the electrochemical responses of these variants with glucose as substrate in the presence of 1,4-BQ, DCPIP, or ferrocene methanol as redox mediators. High catalytic efficiencies (I max app /K M app ) compared to the wild-type POx proved the potential of these variants for future bioelectrocatalytic applications, in biosensors or biofuel cells. Among the variants, L545C showed the most desirable properties as determined kinetically and electrochemically., (Copyright © 2019 The Author(s). Published by Elsevier B.V. All rights reserved.)

Cellobiose dehydrogenase (CDH) is a flavocytochrome with a history of bioelectrochemical research dating back to 1992. During the years, it has been shown to be capable of mediated electron transfer (MET) and direct electron transfer (DET) to a variety of electrodes. This versatility of CDH originates from the separation of the catalytic flavodehydrogenase domain and the electron transferring cytochrome domain. This uncoupling of the catalytic reaction from the electron transfer process allows the application of CDH on many different electrode materials and surfaces, where it shows robust DET. Recent X-ray diffraction and small angle scattering studies provided insights into the structure of CDH and its domain mobility, which can change between a closed-state and an open-state conformation. This structural information verifies the electron transfer mechanism of CDH that was initially established by bioelectrochemical methods. A combination of DET and MET experiments has been used to investigate the catalytic mechanism and the electron transfer process of CDH and to deduce a protein structure comprising of mobile domains. Even more, electrochemical methods have been used to study the redox potentials of the FAD and the haem b cofactors of CDH or the electron transfer rates. These electrochemical experiments, their results and the application of the characterised CDHs in biosensors, biofuel cells and biosupercapacitors are combined with biochemical and structural data to provide a thorough overview on CDH as versatile bioelectrocatalyst., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Isomerization of surface-exposed aspartic acid (Asp) in the complementarity-determining regions of therapeutic proteins could potentially impact their target binding affinity because of the sensitive location, and often requires complex analytical tactics to understand its effect on structure-function and stability. Inaccurate quantitation of Asp-isomerized variants, especially the succinimide intermediate, presents major challenge in understanding Asp degradation kinetics, its stability, and consequently establishing a robust control strategy. As a practical solution to this problem, a comprehensive analytical tool kit has been developed, which provides a solution to fully characterize and accurately quantify the Asp-related product variants. The toolkit offers a combination of 2 steps, an ion-exchange chromatography method to separate and enrich the isomerized variants in the folded structure for structure-function evaluation and a novel focused peptide mapping method to quantify the individual complementarity-determining region isomerization components including the unmodified Asp, succinimide, and isoaspartate. This novel procedure allowed an accurate quantification of each Asp-related variant and a comprehensive assessment of the functional impact of Asp isomerization, which ultimately helped to establish an appropriate control strategy for this critical quality attribute., (Copyright © 2020 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

In this paper we show that conjugation of magnetic nanoparticles (MNPs) with Gemcitabine and/or NucAnt (N6L) fostered their internalization into pancreatic tumor cells and that the coupling procedure did not alter the cytotoxic potential of the drugs. By treating tumor cells (BxPC3 and PANC-1) with the conjugated MNPs and magnetic hyperthermia (43 °C, 60 min), cell death was observed. The two pancreatic tumor cell lines showed different reactions against the combined therapy according to their intrinsic sensitivity against Gemcitabine (cell death, ROS production, ability to activate ERK 1/2 and JNK). Finally, tumors (e.g. 3 mL) could be effectively treated by using almost 4.2 × 10 5 times lower Gemcitabine doses compared to conventional therapies. Our data show that this combinatorial therapy might well play an important role in certain cell phenotypes with low readiness of ROS production. This would be of great significance in distinctly optimizing local pancreatic tumor treatments., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

A new withanolide named as withacoagulin J (1) along with a known withanolide H (2) from Withania coagulans Dunal are reported in this paper. The isolated compounds were elucidated by using 1D-NMR ( 1 H NMR, 13 C NMR) and 2D-NMR including homonuclear (COSY, NOESY) and heteronuclear (HSQC, HMBC); along with Mass spectrometry, UV-Visible and IR spectroscopic techniques. The molecular formula based on Fast-Atom Bombardment Mass Spectrometry [FAB-MS (M + 1)] for 1 and 2 were deduced as C 28 H 37 O 5 and C 28 H 39 O 6 with m/z values 453.2624 and 471.6041, respectively. The quantum mechanical studies of both compounds are based on DFT calculations. The DFT studies show band gaps of 4.86 and 4.83 eV for 1 and 2, respectively. The band gaps of 1 and 2 reflect high stability and resistivity towards oxidation-reduction reactions. The energies of HOMO and LUMO for compound 1 are -6.11 and -1.25 eV and for compound 2: -6.47 and -1.64 eV respectively. Theoretical and experimental FTIR data closely match for both the compounds which support the high accuracy of the computational protocol selection. Other parameters such as bond lengths, bond angles and dihedral angles for both compounds are also studied., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The Mediterranean region is anticipated to be (or, already is) one of the hot spots for climate change, where freshwater ecosystems are under threat from the effects of multiple stressors. Climate change is impacting natural resources and on the functioning of Ecosystem Services. The challenges about modelling climate change impact on water cycle in general and specifically on socio-economic dynamics of the society leads to an exponential amount of results that restrain interpretation and added value of forecasting at local level. One of the main challenges when dealing with climate change projections is the quantification of uncertainties. Modellers might have limited information or understanding from local river catchment management practices and from other disciplines with relevant insights on socio-economic and environmental complex relationship between biosphere and human based activities. Current General Circulation Models cannot fulfil the requirements of high spatial detail required for water management policy. This article reports an innovative transdisciplinary methodology to down scale Climate Change scenarii to river basin level with a special focus on the development of climate change narrative under SSP5-RCP8.5 combination called Myopic scenario and SSP1-RCP4.5 combination called Sustainable scenario. Local Stakeholder participative workshop in the Evrotas river basin provide perception of expected changes on water demand under to two developed scenario narratives., (Copyright © 2018. Published by Elsevier B.V.)

Global change is severely impacting the biosphere that, through ecosystem services, sustains human well-being. Such impacts are expected to increase unless mitigation management actions are implemented. Despite the call from the scientific and political arenas for their implementation, few studies assess the effectiveness of actions on freshwater-related services. Here, by modeling water provisioning, water purification and erosion control under current and future conditions, we assess future trends of service provision with and without mitigation policies. In particular, two different storylines combine multiple climate, land use/land cover and agricultural management scenarios, and represent a pro-efficiency business as usual (myopic storyline) and a future that considers social and environmental sustainability (sustainable storyline). The mentioned services are modeled for the horizon 2050 and in three South European river basins: Ebro, Adige and Sava, which encompass the wide socio-environmental diversity of the region. Our results indicate that Mediterranean basins (Ebro) are extremely vulnerable to global change respect Alpine (Adige) or Continental (Sava) basins, as the Ebro might experience a decrease in water availability up to 40%, whereas the decrease is of only 2-4% in the Adige or negligible in the Sava. However, Mediterranean basins are also more sensitive to the implementation of mitigation actions, which would compensate the drop in water provisioning. Results also indicate that the regulating services of water purification and erosion control will gain more relevance in the future, as both services increased between 4 and 20% in both global change scenarios as a result of the expansion of agricultural and urban areas. Overall, the impact of global change is diverse among services and across river basins in Southern Europe, with the Mediterranean basins as the most vulnerable and the Continental as the least. The implementation of mitigation actions can compensate the impact and therefore deserves full political attention., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Lipidic cubic phase systems (LCPs) are excellent carriers for immobilized enzymes due to their biocompatibility and well-defined nanoporous structure. Lipidic cubic phases act as a convenient matrix to incorporate enzymes and hold them in the vicinity of electrode surfaces in their fully active forms. Corynascus thermophilus cellobiose dehydrogenase (CtCDH) was trapped in a monoolein cubic phase, which increased not only its stability, but also its catalytic performance with both enhanced mediated and direct electron transfer with electrodes. For studies of mediated electron transfer, three mediators with different formal potentials (E°') were employed: horse-heart cytochrome c (cyt c), electron acceptor active with the cytochrome domain of CtCDH, and 2,6-dichlorophenolindophenol (DCPIP) as well as hexaammineruthenium(II) chloride [Ru(NH 3 )Cl 2 ] both electron acceptors with the dehydrogenase domain. Ru(NH 3 )Cl 2 , having the most negative E°' of -0.138V vs. Ag|AgCl at pH7.5, gave a catalytic current for lactose oxidation of 32.10μAcm -2 in MOPS buffer at pH7.5. The process carried out in the same solution but under direct electron conditions transfer resulted in a catalytic current of 9.22μAcm -2 . Electrodes covered with CtCDH in a LCP film retained their catalytic activity after 28days showing a slightly increased current density after 6days., (Copyright © 2017 Elsevier B.V. All rights reserved.)

In this paper, we present an assessment of the sensitivity of groundwater-surface water interactions to climate change in an alluvial aquifer, located in the Ljubljansko polje, Slovenia. The investigation is motivated by a recent assessment of climate change pressures on the water balance in the Sava River Basin (Gampe et al., 2016). The assessment was performed using a comprehensive hydrological modelling approach, which is based on the direct/indirect communication between FEFLOW and WaSiM/MIKE 11. This modelling framework provides a precise simulation of the critical processes in the study domain, which are the main drivers influencing the interactions between precipitation, river water and groundwater under different future climate scenarios. Climate projections were based on the results of the three regional climate models SMHI-RCA4, KNMI-RACMO22E and CLMcom-CCLM4. The results show that there will be higher levels of local precipitation during 2036-2065, the projected river discharge will be larger in the future compared to 2000-2014, and it is unlikely that the Ljubljansko polje will suffer from water scarcity. In addition, amongst the various sections of the Sava River the section between Črnuče and Šentjakob is the one most sensitive to climate change. By running the models under different climate scenarios a deeper insight into aquifer system functioning was obtained. Investigating impacts of climate change on groundwater and interactions between surface water and groundwater on the local scale is a basis for applying such a study on the global scale, which was still not very well investigated., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Protein higher order structure (HOS) is an essential quality attribute to ensure protein stability and proper biological function. Protein HOS characterization is performed during comparability assessments for product consistency as well as during forced degradation studies for structural alteration upon stress. Circular dichroism (CD) spectroscopy is a widely used technique for measuring protein HOS, but it remains difficult to assess HOS with a high degree of accuracy and precision. Moreover, once spectral changes are detected, interpreting the differences in terms of specific structural attributes is challenging. Spectral normalization by the protein concentration remains one of the largest sources of error and reduces the ability to confidently detect differences in CD spectra. This work develops a simple method to enhance the precision of the CD spectral measurements through normalization of the CD spectra by the protein concentration determined directly from the CD measurement. This method is implemented to successfully detect small CD spectral changes in multiple forced degradation studies as well as comparability assessments during biologics drug development. Furthermore, the interpretation of CD spectral changes in terms of HOS differences are provided based on orthogonal data in conjunction with structural insights gained through in silico homology modeling of the protein structure., (Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

To develop meaningful land use scenarios, drivers that affect changes in the landscape are required. In this study, driving factors that influence farmers to change crops on their farm were determined. A questionnaire was administered to four independent groups of farmers who identified and ranked influencing factors pertaining to their choices of crops. The farmers were located in two mid-latitude agricultural watersheds (in Germany and Canada). The ranked influencing factors were used to develop a "farmer driven" scenario to 2040 in both watersheds. Results showed that the most important influencing factors for farmers to change crops were the "economic return of the crop" and "market factors". Yet, when the drivers of crop land use change were grouped into two categories of "financial" and "indirectly-related financial" factors, the "financial" factors made up approximately half of the influencing factors. For some responses, the "indirectly-related financial" factors (i.e. "access to farm equipment", the "farm experience", and "climate") ranked higher than or just as high as the financial factors. Overall, in the four farmer groups the differences between the rankings of the influencing factors were minor, indicating that drivers may be transferable between farms if the farmers are full-time and the farming regions have comparable growing seasons, access to markets, similar technology, and government programs for farm income. In addition to the "farmer driven" scenario, a "policy driven" scenario was derived for each watershed based only on available information on the financial incentives provided to farmers (i.e. agricultural subsidies, income support, crop insurance). The influencing factors ranked by the farmers provided in-depth information that was not captured by the "policy driven" scenario and contributed to improving predictions for crop land use development. This straight-forward method to rank qualitative data provided by farmers can easily be replicated in other watersheds to improve environmental impact modelling., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Multiple abiotic stressors affect the ecological status of water bodies. The status of waterbodies in the Ebro catchment (NE Spain) is evaluated using the biological quality elements (BQEs) of diatoms, invertebrates and macrophytes. The multi-stressor influence on the three BQEs was evaluated using the monitoring dataset available from the catchment water authority. Nutrient concentrations, especially total phosphorus (TP), affected most of the analyzed BQEs, while changes in mean discharge, water temperature, or river morphology did not show significant influences. Linear statistical models were used to evaluate the change of water bodies' ecological status under different combinations of future socioeconomic and climate scenarios. Changes in land use, rainfall, water temperature, mean discharge, TP and nitrate concentrations were modeled according to the future scenarios. These revealed an evolution of the abiotic stressors that could lead to a general decrease in the ecosystem quality of water bodies within the Ebro catchment. This deterioration was especially evidenced on the diatoms and invertebrate biological indices, mainly because of the foreseen increase in TP concentrations. Water bodies located in the headwaters were seen as the most sensitive to future changes., (Copyright © 2018 The Author(s). Published by Elsevier B.V. All rights reserved.)

The objective of this study is to provide spatially distributed water use maps at a high spatial and thematic resolution as detailed input data for further modelling purposes. The maps were derived on the basis of official water statistics and land use maps to represent the current conditions in four European river basins affected by water scarcity (Adige, Ebro, Evrotas and Sava) and allow setting up reference scenarios. The present land use distribution was modelled based on CORINE data with the land use change model iCLUE. Modelling the land use allows to create dynamic land and water use maps adapted to the needs of eventual scenario analyses compared to using only observed land use maps. The availability of several CORINE datasets allowed calibrating and validating the results of the iCLUE model carrying out a three map comparison. Sectoral water uses were attributed to different land use classes and by this means located in space. Both the location and the magnitude of urban and agricultural water uses can be derived from the final maps. The created maps together with the corresponding land use data provide a coherent set of information crucial to most environmental modelling activities and often missing at this spatial and thematic resolution. This work also aims at visualizing and validating the water use statistics provided by official institutions such as the River Basin Management Plans. The results show that in some cases they are not consistent and underline the importance of harmonised data collection regarding water statistics, as otherwise comparisons within one study area and with others are hampered. This study is embedded in the EU-FP7 GLOBAQUA project which analyses the effects of stressors, such as changes of land and water use, on aquatic ecosystems in areas suffering water scarcity., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Sustainable water basin management requires characterization of flow regime in river networks impacted by anthropogenic pressures. Flow regime in ungauged catchments under current, future, or natural conditions can be assessed with hydrological models. Developing hydrological models is, however, resource demanding such that decision makers might revert to models that have been developed for other purposes and are made available to them ('off-the-shelf' models). In this study, the impact of epistemic uncertainty of flow regime indicators on flow-ecological assessment was assessed at selected stations with drainage areas ranging from about 400 to almost 90,000km 2 in four South European basins (Adige, Ebro, Evrotas and Sava). For each basin, at least two models were employed. Models differed in structure, data input, spatio-temporal resolution, and calibration strategy, reflecting the variety of conditions and purposes for which they were initially developed. The uncertainty of modelled flow regime was assessed by comparing the modelled hydrologic indicators of magnitude, timing, duration, frequency and rate of change to those obtained from observed flow. The results showed that modelled flow magnitude indicators at medium and high flows were generally reliable, whereas indicators for flow timing, duration, and rate of change were affected by large uncertainties, with correlation coefficients mostly below 0.50. These findings mirror uncertainty in flow regime indicators assessed with other methods, including from measured streamflow. The large indicator uncertainty may significantly affect assessment of ecological status in freshwater systems, particularly in ungauged catchments. Finally, flow-ecological assessments proved very sensitive to reference flow regime (i.e., without anthropogenic pressures). Model simulations could not adequately capture flow regime in the reference sites comprised in this study. The lack of reliable reference conditions may seriously hamper flow-ecological assessments. This study shows the pressing need for improving assessment of natural flow regime at pan-European scale., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

Sequence variants of a monoclonal antibody resulting from incomplete processing of signal peptide were identified and characterized using multiple mass spectrometry platforms and reverse phase chromatography. Detection and quantification of these variants by three LC/MS platforms were assessed. Quantification was also performed by mass spectrometric analysis of the subunits of the antibody generated by reduction and IdeS proteolysis. Peptide mapping with LC/MS/MS detection was used to quantify and confirm the identities of signal peptide sequence variants. Although quantification of the signal peptide variants thru mass spectrometry approaches is system dependent, our data revealed the results are close to the values determined by chromatographic separation with UV detection. Each of the methods have proven effective in demonstrating the consistency of signal peptide variants levels across the manufacture history of the antibody., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Nowadays, nanomaterials with remarkable antibacterial activity and low cytotoxicity attract much interest in research. By considering the antibacterial activity of Ag and graphene oxide (GO), the Ag-RGO nanocomposite was prepared by a one-pot and facile technique and it was used to evaluate its antibacterial activity and cytotoxicity against Escherichia coli and glioblastoma cancer cells (U87MG), respectively. The antibacterial activity was studied by micro-dilution and colony counting methods to investigate cell viability. The viability of glioblastoma cells was determined using MTT assay. Since MIC and MBC values of the nanocomposite are 20 and 40μg/mL, respectively, it acts as a bactericidal agent. The antibacterial properties of nanocomposite are time and concentration dependent. The kinetics and mechanism of the antibacterial activity of the nanocomposite were investigated. The antibacterial activity for Ag-RGO nanocomposite is induced by capturing-killing process. From the results, we concluded that Ag-RGO nanocomposite can simultaneously induce apoptosis. Our results bring up a new plan for the use of silver nanoparticles in the form of nanocomposite with reduced graphene oxide in antibacterial applications. Also, Ag-RGO nanocomposite can reduce the viability of U87MG in a dose dependent manner which may show its anticancer potential., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far., Methods: To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used., Results: HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation., Conclusions: Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH., General Significance: The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The contribution illustrates an integrated assessment framework aimed at evaluating the relationships between multiple pressures and water body status for the purposes of river basin management. The framework includes the following steps. (1) Understanding how the different pressures affect the status of water bodies. This entails the characterization of biophysical state variables and the definition of a causal relationship between pressures and status. Therefore this step involves interaction between experts bearing ecological understanding and experts providing models to represent the effect of pressures. (2) Identifying the relevant pressures to be addressed through appropriate measures to improve the status of water bodies. (3) Evaluating reduction targets for the relevant pressures identified in a river basin, by weighting the effort associated to reducing individual pressures and the potential benefits in terms of water body status. (4) Designing management measures through a creative process and political discussion of alternative options, balancing costs, benefits and effectiveness based on engineering and economic analysis. (5) Simulating scenarios of implementation of a programme of measures in order to check their effectiveness and robustness against climate and land use change. We discuss the five steps of the assessment framework, and particularly the interaction between science and policy at the different stages. We review the assessment tools required at each step and, for setting optimal pressure reduction targets (step 3), we propose and illustrate a simplified multicriteria approach based on semi-quantitative assessment, which produces frontiers of optimal trade-offs between effort spent on measures, and achievements., (Copyright © 2016. Published by Elsevier B.V.)

Climate change will likely increase pressure on the water balances of Mediterranean basins due to decreasing precipitation and rising temperatures. To overcome the issue of data scarcity the hydrological relevant variables total runoff, surface evaporation, precipitation and air temperature are taken from climate model simulations. The ensemble applied in this study consists of 22 simulations, derived from different combinations of four General Circulation Models (GCMs) forcing different Regional Climate Models (RCMs) and two Representative Concentration Pathways (RCPs) at ~12km horizontal resolution provided through the EURO-CORDEX initiative. Four river basins (Adige, Ebro, Evrotas and Sava) are selected and climate change signals for the future period 2035-2065 as compared to the reference period 1981-2010 are investigated. Decreased runoff and evaporation indicate increased water scarcity over the Ebro and the Evrotas, as well as the southern parts of the Adige and the Sava, resulting from a temperature increase of 1-3° and precipitation decrease of up to 30%. Most severe changes are projected for the summer months indicating further pressure on the river basins already at least partly characterized by flow intermittency. The widely used Falkenmark indicator is presented and confirms this tendency and shows the necessity for spatially distributed analysis and high resolution projections. Related uncertainties are addressed by the means of a variance decomposition and model agreement to determine the robustness of the projections. The study highlights the importance of high resolution climate projections and represents a feasible approach to assess climate impacts on water scarcity also in regions that suffer from data scarcity., (Copyright © 2016. Published by Elsevier B.V.)

The reliable assessment of water quality through water quality monitoring programs (WQMPs) is crucial in order for decision-makers to understand, interpret and use this information in support of their management activities aiming at protecting the resource. The challenge of water quality monitoring has been widely addressed in the literature since the 1940s. However, there is still no generally accepted, holistic and practical strategy to support all phases of WQMPs. The purpose of this paper is to report on the use cases a watershed manager has to address to plan or optimize a WQMP from the challenge of identifying monitoring objectives; selecting sampling sites and water quality parameters; identifying sampling frequencies; considering logistics and resources to the implementation of actions based on information acquired through the WQMP. An inventory and critique of the information, approaches and tools placed at the disposal of watershed managers was proposed to evaluate how the existing information could be integrated in a holistic, user-friendly and evolvable solution. Given the differences in regulatory requirements, water quality standards, geographical and geological differences, land-use variations, and other site specificities, a one-in-all solution is not possible. However, we advance that an intelligent decision support system (IDSS) based on expert knowledge that integrates existing approaches and past research can guide a watershed manager through the process according to his/her site-specific requirements. It is also necessary to tap into local knowledge and to identify the knowledge needs of all the stakeholders through participative approaches based on geographical information systems and adaptive survey-based questionnaires. We believe that future research should focus on developing such participative approaches and further investigate the benefits of IDSS's that can be updated quickly and make it possible for a watershed manager to obtain a timely, holistic view and support for every aspect of planning and optimizing a WQMP., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Water scarcity and water security are linked, not only through the direct effects of water shortages on each water users' access to water, but also because of water conflicts generated. Climate change is predicted to raise temperatures in the Mediterranean region and reduce rainfall, leading to a reduction in water yield and possibly worsening the situation of water resource shortages that Mediterranean regions are already experiencing. In its dissemination strategy, the EU FP7 CLIMB project addressed water security threats through an analysis of water uses and water use rivalries within a few target catchments distributed over the Mediterranean region. The present work explores whether climate change is locally perceived by stakeholders (water users and managers) as a key issue for their water uses and water security. Individual interviews, meetings, and compilation of questionnaires were conducted at five sites located in the Mediterranean region. The methodology permitted an analysis of water use and its evolution in the water management context, an identification of the state of awareness of local stakeholders and of the pressures on water use and water use rivalries, and a prioritization of water uses. Currently, the main response to increasing water demand in the Mediterranean region, while not yet considering climate change as a driving force, is a progressive externalization of water resources, with limits represented by national borders and technological possibilities. Overall, 'climate change' was not mentioned by stakeholders during both interviews and in answers to the questionnaires. Even the prospect of decreasing precipitation was not considered a relevant or threatening issue in the coming 20years. This confirms the need to continue all efforts to disseminate the state of knowledge on climate change impacts in the Mediterranean region, such as water scarcity, especially to local water managers, as initiated by various research programs of the European Commission., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In the Mediterranean region, particularly in the Gaza strip, an increased risk of drought is among the major concerns related to climate change. The impacts of climate change on water availability, drought risk and food security can be assessed by means of hydro-climatological modeling. However, the region is prone to severe observation data scarcity, which limits the potential for robust model parameterization, calibration and validation. In this study, the physically based, spatially distributed hydrological model WaSiM is parameterized and evaluated using satellite imagery to assess hydrological quantities. The Triangle Method estimates actual evapotranspiration (ETR) through the Normalized Difference Vegetation Index (NDVI) and land surface temperature (LST) provided by Landsat TM imagery. So-derived spatially distributed evapotranspiration is then used in two ways: first a subset of the imagery is used to parameterize the irrigation module of WaSiM and second, withheld scenes are applied to evaluate the performance of the hydrological model in the data scarce study area. The results show acceptable overall correlation with the validation scenes (r=0.53) and an improvement over the usual irrigation parameterization scheme using land use information exclusively. This model setup is then applied for future drought risk assessment in the Gaza Strip using a small ensemble of four regional climate projections for the period 2041-2070. Hydrological modeling reveals an increased risk of drought, assessed with an evapotranspiration index, compared to the reference period 1971-2000. Current irrigation procedures cannot maintain the agricultural productivity under future conditions without adaptation., (Copyright © 2015 Elsevier B.V. All rights reserved.)

According to current climate projections, Mediterranean countries are at high risk for an even pronounced susceptibility to changes in the hydrological budget and extremes. These changes are expected to have severe direct impacts on the management of water resources, agricultural productivity and drinking water supply. Current projections of future hydrological change, based on regional climate model results and subsequent hydrological modeling schemes, are very uncertain and poorly validated. The Rio Mannu di San Sperate Basin, located in Sardinia, Italy, is one test site of the CLIMB project. The Water Simulation Model (WaSiM) was set up to model current and future hydrological conditions. The availability of measured meteorological and hydrological data is poor as it is common for many Mediterranean catchments. In this study we conducted a soil sampling campaign in the Rio Mannu catchment. We tested different deterministic and hybrid geostatistical interpolation methods on soil textures and tested the performance of the applied models. We calculated a new soil texture map based on the best prediction method. The soil model in WaSiM was set up with the improved new soil information. The simulation results were compared to standard soil parametrization. WaSiMs was validated with spatial evapotranspiration rates using the triangle method (Jiang and Islam, 1999). WaSiM was driven with the meteorological forcing taken from 4 different ENSEMBLES climate projections for a reference (1971-2000) and a future (2041-2070) times series. The climate change impact was assessed based on differences between reference and future time series. The simulated results show a reduction of all hydrological quantities in the future in the spring season. Furthermore simulation results reveal an earlier onset of dry conditions in the catchment. We show that a solid soil model setup based on short-term field measurements can improve long-term modeling results, which is especially important in ungauged catchments., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A new isoflavone 7-hydroxy 3'-methoxyisoflavone (1) is isolated from the seeds of Indigofera heterantha. The structure of this new compound was established using spectroscopic techniques such as ID, 2D NMR, and mass spectrometry. Density functional theory calculations are carried out for the first time for geometric, electronic and spectroscopic properties of 1 (isoflavone). DFT calculations have been performed at B3LYP/6-311G(d,p) level of theory for obtaining geometric and spectroscopic properties of compound 1. The simulated vibrational spectrum of compound 1 at B3LYP/6-311G(d,p) shows nice correlation with the experimental IR spectrum after a scaling factor of 0.973. (1)H and (13)C NMR chemical shifts were calculated using Cramer's re-parameterized function WP04 at 6-311G(d,p) basis set, and show nice correlation with the experimental data. Four conformers were considered for NMR chemical shift calculations. Electronic properties such as band gap, Ionization potential and electron affinities were also simulated for the first time; however, no comparison could be made with the experiment., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The phytochemical examination of chloroform soluble fraction (FX2) of methanolic extract of bark of Millettia ovalifolia yielded a new flavonoid; 7-(4-methoxyphenyl)-9H-furo [2,3-f]chromen-9-one (1). Compound 1 is characterized by spectroscopic analytical techniques such as UV, IR, 1D, 2D NMR spectroscopy, and mass spectrometry. A theoretical model is also developed for obtaining geometric, electronic and spectroscopic properties of 1. The geometry optimization and harmonic vibration simulations have been carried out at B3LYP/6-31G(d,p). The vibrational spectrum of compound 1 shows nice correlation with the experimental IR spectrum, through a scaling factor of 0.9613. (1)H and (13)C NMR chemical shifts are simulated using Cramer's re-parameterized function WP04 at 6-31G(d,p) basis set, and correlate nicely with the experimental chemical shifts., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Unspecific peroxygenase (UPO) is a highly efficient biocatalyst with a peroxide dependent monooxygenase activity and many biotechnological applications, but the absence of suitable heterologous expression systems has precluded its use in different industrial settings. Recently, the UPO from Agrocybe aegerita was evolved for secretion and activity in Saccharomyces cerevisiae [8]. In the current work, we describe a tandem-yeast expression system for UPO engineering and large scale production. By harnessing the directed evolution process in S. cerevisiae, the beneficial mutations for secretion enabled Pichia pastoris to express the evolved UPO under the control of the methanol inducible alcohol oxidase 1 promoter. Whilst secretion levels were found similar for both yeasts in flask fermentation (∼8mg/L), the recombinant UPO from P. pastoris showed a 27-fold enhanced production in fed-batch fermentation (217mg/L). The P. pastoris UPO variant maintained similar biochemical properties of the S. cerevisiae counterpart in terms of catalytic constants, pH activity profiles and thermostability. Thus, this tandem-yeast expression system ensures the engineering of UPOs to use them in future industrial applications as well as large scale production., (Copyright © 2015 Elsevier Inc. All rights reserved.)