bluuebunny/crossref_metadata_2025 huggingface dataset:
Development, Screening, and Validation of Camelid‐Derived Nanobodies for Neuroscience Research
Authors: Gavira‐O'Neill Clara E. , Dong Jie‐Xian , Trimmer James S.
Published: November 2020

Nanobodies (nAbs) are recombinant antigen‐binding variable domain fragments obtained from heavy‐chain‐only immunoglobulins. Among mammals, these are unique to camelids (camels, llamas, alpacas, etc.). Nanobodies are of great use in biomedical research due to their efficient folding and stability under a variety of conditions, as well as their small size. The latter characteristic is particularly important for nAbs used as immunolabeling reagents, since this can improve penetration of cell and tissue samples compared to conventional antibodies, and also reduce the gap distance between signal and target, thereby improving imaging resolution. In addition, their recombinant nature allows for unambiguous definition and permanent archiving in the form of DNA sequence, enhanced distribution in the form of sequences or plasmids, and easy and inexpensive production using well‐established bacterial expression systems, such as the IPTG induction method described here. This article will review the basic workflow and process for developing, screening, and validating novel nAbs against neuronal target proteins. The protocols described make use of the most common nAb development method, wherein an immune repertoire from an immunized llama is screened via phage display technology. Selected nAbs can then be taken through validation assays for use as immunolabels or as intrabodies in neurons. © 2020 Wiley Periodicals LLC.

This article was corrected on 26 June 2021. See the end of the full text for details.

Basic Protocol 1: Total RNA isolation from camelid leukocytes

Basic Protocol 2: First‐strand cDNA synthesis; VHH and VH repertoire PCR

Basic Protocol 3: Preparation of the phage display library

Basic Protocol 4: Panning of the phage display library

Basic Protocol 5: Small‐scale nAb expression

Basic Protocol 6: Sequence analysis of selected nAb clones

Basic Protocol 7: Nanobody validation as immunolabels

Basic Protocol 8: Generation of nAb‐pEGFP mammalian expression constructs

Basic Protocol 9: Nanobody validation as intrabodies

Support Protocol 1: ELISA for llama serum testing, phage titer, and screening of selected clones

Support Protocol 2: Amplification of helper phage stock

Support Protocol 3: nAb expression in amber suppressor E. coli bacterial strains
Use of wild Pennisetum species for improving biotic and abiotic stress tolerance in pearl millet
Authors: Sharma Shivali , Sharma Rajan , Pujar Mahesh , Yadav Devvart , Yadav Yashpal , Rathore Abhishek , Mahala Rajendra Singh , Singh Indra , Verma Yogendra , Deora Virendra Singh , Vaid Bhupesh , Jayalekha Ayyathan Kakkadan , Gupta Shashi Kumar
Published: November 2020

Pearl millet [Pennisetum glaucum (L.) R. Br.] is one of the world's hardiest warm‐season cereal crop cultivated for food and animal feed in the semi‐arid tropics of Asia and Africa. This crop faces terminal drought during rainy and flowering‐stage heat stress during summer seasons. Blast is emerging as a serious threat affecting its production and productivity in India. Using wild P. violaceum (Lam) Rich. and pearl millet cultivars, prebreeding populations were developed following backcross method. These populations were evaluated in target ecologies in India at three locations during the 2018 summer season for flowering‐stage heat stress and at two locations during the 2018 rainy season for terminal drought stress. A total 18 introgression lines (ILs) from Population (Pop) 3 exhibited improved seed set under high heat stress vs. the cultivated parent, whereas no IL was better than the cultivated parent in Pop 4. Under rainfed conditions at Hisar and Bawal, India, 19 ILs from Pop 3 and 16 ILs from Pop 4 showed significantly higher dry fodder yield than the cultivated parents. Further, screening of ILs for five diverse pathotype isolates—Pg 45, Pg 138, Pg 186, Pg 204, and Pg 232—of blast resulted in the identification of resistant ILs. Use of these promising ILs in breeding programs will assist in developing new varieties and hybrids with improved tolerance to biotic and abiotic stresses. The study indicated the genetic differences between the parents involved in crossing and also highlighted the importance of precise phenotyping of wild species for target trait prior to use in prebreeding work.
Investigations into the stable isotope ratios of 1‐phenyl‐2‐propanone
Authors: Lansdown Kate , Jolliffe Katrina A. , Salouros Helen
Published: November 2020

Stable isotope ratio mass spectrometry (IRMS) can be used to determine the precursor and precursor origin of methylamphetamine drug samples. Previous work has shown that methylamphetamine samples can be distinguished as derived from different sources of (pseudo)ephedrine or phenyl‐2‐propanone (P2P) through the use of IRMS alongside conventional chemical profiling techniques. To date, limited research has been conducted to investigate whether methylamphetamine samples of differing P2P origins can be distinguished through drug profiling. This was investigated by synthesising methylamphetamine in‐house in a three‐step process. Two ‘preprecursors’ were used in this study, phenylacetic acid (PAA) and α‐phenylacetoacetonitrile (α‐PAAN). Using a combination of profiling techniques, it was found that methylamphetamine samples of PAA preprecursor origin and methylamphetamine samples of α‐PAAN preprecursor origin can be distinguished.
Annual banned‐substance review: Analytical approaches in human sports drug testing 2019/2020
Authors: Thevis Mario , Kuuranne Tiia , Geyer Hans
Published: November 2020

Analytical chemistry‐based research in sports drug testing has been a dynamic endeavor for several decades, with technology‐driven innovations continuously contributing to significant improvements in various regards including analytical sensitivity, comprehensiveness of target analytes, differentiation of natural/endogenous substances from structurally identical but synthetically derived compounds, assessment of alternative matrices for doping control purposes, and so forth. The resulting breadth of tools being investigated and developed by anti‐doping researchers has allowed to substantially improve anti‐doping programs and data interpretation in general. Additionally, these outcomes have been an extremely valuable pledge for routine doping controls during the unprecedented global health crisis that severely affected established sports drug testing strategies. In this edition of the annual banned‐substance review, literature on recent developments in anti‐doping published between October 2019 and September 2020 is summarized and discussed, particularly focusing on human doping controls and potential applications of new testing strategies to substances and methods of doping specified the World Anti‐Doping Agency's 2020 Prohibited List.
The association between leptin and weight maintenance outcome in anorexia nervosa
Authors: Kim Youngjung , Hersch Jonathan , Bodell Lindsay P. , Schebendach Janet , Hildebrandt Tom , Walsh B. Timothy , Mayer Laurel E. S.
Published: November 2020

Objective

Relapse after weight restoration in anorexia nervosa (AN) is a critical problem. Higher body fat percentage after weight gain has been shown to predict better weight maintenance outcome. Leptin, a fat‐derived hormone, has been associated with progress during weight gain, but its association with weight maintenance is unknown. This study aims to determine whether leptin levels after weight restoration in AN are associated with weight maintenance.

Method

Participants were 41 women with AN hospitalized for inpatient treatment. Participants were evaluated 2–4 weeks after weight restoration to body mass index (BMI) ≥ 19.5 kg/m2 for plasma leptin and body composition. Weight maintenance outcome was defined by whether a participant maintained a BMI of at least 18.5 kg/m2 at the end of 1 year following hospital discharge.

Results

Twenty (48.8%) out of 41 patients maintained their weight at 1 year. Percent body fat and leptin were significantly higher in the group who maintained weight (body fat, p = .004, Hedges' g = 0.944; log‐leptin, p = .010, Hedges' g = 0.821), but there were no differences in predischarge BMI, duration of illness, and duration of amenorrhea. Using regression modeling, only higher log‐leptin (pWald = .021) and percent body fat (pWald = .010), as well as fat‐adjusted leptin (pWald = .029), independently predicted weight maintenance at 1 year.

Discussions

Our findings suggest that for acutely‐weight restored women with AN, higher predischarge leptin measurements are associated with better outcome in the year following treatment. Prospective studies examining leptin as well as other parameters of metabolic health could offer insights into biomarkers that may improve clinical outcomes.
Investigating the role of matrix habitat use in determining avian area‐sensitivity
Authors: Roberts H. Patrick , King David I.
Published: November 2020

The absence of some species from small habitat patches has long posed a challenge for conservationists, yet the underlying mechanisms that cause this “area‐sensitivity” remain poorly understood. Capacity of a species to extend their activities into the surrounding matrix habitat represents one potential determinant of area‐sensitivity. Species may be able to occupy smaller patches if they can utilize matrix habitat beyond patch boundaries, whereas area‐sensitive species may be restricted to larger patches due to their inability to utilize the surrounding matrix. We investigated the potential role of matrix utilization in determining area‐sensitivity by mapping the movements of two shrubland‐obligate passerines with contrasting patch area requirements in shrub‐dominated forest openings ranging in area by nearly an order of magnitude. Our findings were consistent with our predictions; the less area‐sensitive chestnut‐sided warbler (Setophaga pensylvanica) exhibited greater use of matrix habitat than the highly area‐sensitive prairie warbler (S. discolor). Furthermore, chestnut‐sided warblers that occupied smaller openings used mature forest more than conspecifics in larger patches, yet forest use by prairie warblers was unrelated to opening size. Chestnut‐sided warblers foraged as frequently in mature forest as within shrubland, whereas prairie warblers foraged significantly more in openings compared to forest. The findings of this study suggest that the ability or inclination of a species to utilize surrounding matrix habitat explains at least some of the observed variation in area‐sensitivity in songbirds and potentially other taxa.
Do I really want to engage in contact? Volition as a new dimension of intergroup contact
Authors: Bagci Sabahat C. , Husnu Shenel , Turnuklu Abbas , Tercan Mustafa
Published: November 2020

Despite burgeoning evidence for the effectiveness of intergroup contact on prejudice reduction, relatively less is known about how much individuals' internal control and active choice is involved in the formation of intergroup contact. Through four correlational studies (Ntotal = 1043, Study 1 in Cyprus and Studies 2–4 in Turkey), we investigated the construct of ‘contact volition’—the extent to which one engages in intergroup contact deliberately. Studies 1–3 showed that intergroup contact perceived as primarily intentional and pursued by the active choice of the self (volitional contact) was associated with greater intimacy and positivity and thereby more positive attitudes, compared to contact that is primarily based on external/situational conditions (contingent contact). Study 4 showed that contact quantity was associated with more positive attitudes and behavioral tendencies through greater intimacy and positivity, only when contact was volitional. Findings contribute to the recently emerging research literature on motivated contact behavior that can help maximize the effectiveness of contact.
An integrated microfluidics for assessing the anti‐aging effect of caffeic acid phenethylester in Caenorhabditis elegans
Authors: Zhang Beibei , Zhuang Liping , Sun Duanping , Li Yinbao , Chen Zuanguang
Published: November 2020

Aging is a fundamental and fascinating process. Anti‐aging research tried to find the mysteries about the human lifespan. To investigate the longevity‐extending role of caffeic acid phenethylester (CAPE) and reveal the possible regulation mechanism, the long‐term cultivation under well‐defined environments, real‐time monitoring, and live imaging is highly desired. In this paper, a well‐designed microfluidic device was proposed to analyze the anti‐aging effect of CAPE in Caenorhabditis elegans. With the combined use of multiple functional units, including micro‐pillar, worm responder, a branching network of distribution channels, and microchambers, the longitudinal measurements of the exact number of worms throughout the whole lifespans is possible. Meanwhile, the brief cooling function of temperature‐controllable system can achieve temporary and repeated immobilization of nematodes for fluorescence imaging. Our research data showed that CAPE can increase the survival of worms under normal and stress condition, including heat stress and paraquat‐induced oxidative stress. The further studies revealed the anti‐aging mechanism of CAPE. This proposed strategy and device would be a useful platform to facilitate future anti‐aging studies and the finding of new lead compounds.
Temperature‐dependent inactivation and catalysis rates of plant‐based ureases for engineered biomineralization
Authors: Feder Marnie J , Akyel Arda , Morasko Vincent J , Gerlach Robin , Phillips Adrienne J
Published: November 2020

Engineered (bio)mineralization uses the enzyme urease to catalyze the hydrolysis of urea to promote carbonate mineral precipitation. The current study investigates the influence of temperature on ureolysis rate and degree of inactivation of plant‐sourced ureases over a range of environmentally relevant temperatures. Batch experiments at 30°C demonstrated that jack bean meal (JBM) has a 1.7 to 56 times higher activity (844 μmol urea hydrolyzed g−1 JBM min−1) than the other tested plant‐sourced ureases (soybean, pigeon pea and cottonseed). Hence, ureolysis and enzyme inactivation rates were evaluated for JBM at temperatures between 20°C and 80°C. A combined first‐order urea hydrolysis and first‐order enzyme inactivation model described the inactivation of urease over the investigated range of temperatures. The temperature‐dependent rate coefficients (kurea) increased with temperature and ranged from 0.0018 at 20°C to 0.0249 L g−1 JBM min−1 at 80°C; JBM urease became ≥50% inactivated in as little as 5.2 minutes at 80°C and in as long as 2238 minutes at 50°C. The combined urea hydrolysis kinetics and enzyme inactivation model provides a mathematical relationship useful for the design of biomineralization technologies and can be incorporated into reactive transport models.
Strategies for Alleviating Electrode Expansion of Graphite Electrodes in Sodium‐Ion Batteries Followed by In Situ Electrochemical Dilatometry
Authors: Escher Ines , Kravets Yuliia , Ferrero Guillermo A. , Goktas Mustafa , Adelhelm Philipp
Published: November 2020

The electrochemical intercalation/deintercalation of solvated sodium ions into graphite is a highly reversible process, but leads to large, undesired electrode expansion/shrinkage (“breathing”). Herein, two strategies to mitigate the electrode expansion are studied. Starting with the standard configuration (−) sodium | diglyme (2G) electrolyte | graphite (poly(vinylidene difluoride) (PVDF) binder) (+), the PVDF binder is first replaced with a binder made of the sodium salt of carboxymethyl cellulose (CMC). Second, ethylenediamine (EN) is added to the electrolyte solution as a co‐solvent. The electrode breathing is followed in situ (operando) through electrochemical dilatometry (ECD). It is found that replacing PVDF with CMC is only effective in reducing the electrode expansion during initial sodiation. During cycling, the electrode breathing for both binders is comparable. Much more effective is the addition of EN. The addition of 10 v/v EN to the diglyme electrolyte strongly reduces the electrode expansion during the initial sodiation (+100% with EN versus +175% without EN) as well as the breathing during cycling. A more detailed analysis of the ECD signals reveals that solvent co‐intercalation temporarily leads to pillaring of the graphite lattice and that the addition of EN to 2G leads to a change in the sodium storage mechanism.
Polyethylene film modification using polylactic acid ‐ Starch additives and study ionizing radiation effect onto aging properties
Authors: Maziad Nabila A. , Abdel Ghaffar Ashraf M. , Ali Hussein E.
Published: November 2020

Modification of low density polyethylene (LDPE) films by blending with Polylactic acid (PLA) and Starch (St) were carried out. Also, the study describes the comparison of two curing methods one by thermal curing at 70°C and the other is radiation curing at dose 20 kGy for the bioblend film composed of LDPE/PLA/St with composition (98%:1%:1% wt%). The efficiency of modification was determined by using different devices such as Fourier transform infrared (FTIR), mechanical, TGA, and XRD. In addition, the properties of the different bioblends were compared with other synthetic polymers such as commercial LDPE and blank LDPE. The results show that the improved properties of radiation cured method at dose 20 kGy than thermal treatment method at 70°C. The radiation cured bioblend film show good thermal stability either shown by the TGA results or against heating aging from ambient temperature up to 60°C than thermally treated blend film at 70°C. The radiation cured bioblend film found to be stable against UV‐ exposure with increasing time of exposure up to 30 min at higher wavelength (λ = 320 nm), which simulate ultraviolet radiation from sunshine. The modified properties of prepared radiation cured bioblend film are due to increase in interfacial adhesion bond between LDPE, PLA, and Starch by effect of irradiation dose at 20 kGy. The modified radiation cured bioblend films at 20 kGy can be applied in different fields such as packaging due to improvement in chemical, thermal, and mechanical properties that could partially replace the traditional petrochemical plastics.
Incorporation of heteroatoms into reticulated vitreous carbon foams derived from sucrose to improve its energy storage performance
Authors: Vazquez‐Samperio Juvencio , Acevedo‐Peña Próspero , Guzmán‐Vargas Ariel , Reguera Edilso , Cordoba‐Tuta Elcy
Published: November 2020

Herein, a binder‐free 3D reticulated carbon doped with N, F, and B is proposed as an excellent choice to be used as an electrode in volumetric supercapacitors for stationary devices. The carbonaceous electrode was obtained by pyrolysis (N2 atmosphere) of sucrose resin modified with precursors of heteroatoms doping, impregnated on the polyurethane foam used as a template. Different techniques such as SEM, Raman, XPS, and FTIR were employed to characterize the properties of RVC. Cyclic Voltammetry, Galvanostatic charge and discharge curves, and EIS were used to evaluate the energy storage performance of the foams. The electrochemical evaluation in acidic medium shows that the composition and chemical state of doping elements in the RVC determine their capacitance. RVC‐N electrode exhibits the best performance among the foams here synthetized registering volumetric capacitances of 3.57, 3.20, 2.25, 1.45, and 0.63 F cm−3 at 5, 10, 20, 40, and 80 mV s−1, respectively. Further, a symmetric supercapacitor was assembled by using RVC‐N as a positive and negative electrode. The device shows volumetric capacitances of 3.82, 1.4, 0.57, 0.47, 0.37, 0.24, and 0.15 F cm−3 at 0.5, 1, 2, 4, 6, 18 y 35 mA cm−3 inside a voltage window of 1.4 V. Moreover, the device retained the 80% of its initial capacitance after 10 000 cycles.
Another Chance for Classic AFCs? Experimental Investigation of a Cost‐Efficient Unitized Regenerative Alkaline Fuel Cell, Using Platinum‐Free Gas Diffusion Electrodes
Authors: Wagner E. , Kohnke H.‐J.
Published: November 2020

The big challenge of global phase‐down of carbon dioxide emissions leads us to alternatives in storing electric energy from renewable sources. For the worldwide use of batteries and fuel cells very high amounts of precious electrode materials are needed. Batteries require rare earth metals and environmentally harmful Lithium, PEM fuel cells require noble platinum and iridium. Thus, an economical and eco‐friendly alternative to lithium‐ion‐batteries should be found, especially for the use in domestic homes. In the on‐hand work, a well‐known type of fuel cell is revised and considered in the new context of global material usage. Therefore, an alkaline fuel cell with the classic design of an electrolyte gap in‐between the electrodes is observed. Gas‐diffusion‐electrodes are used, based on Raney‐nickel. Results are given for different catalysts: Raney‐nickel, Raney‐silver, manganic‐oxide, carbon and ruthenium. For the analysis current‐voltage‐characteristics, chronopotentiometry and SEM micrographs are used. The results are discussed in comparison to a unitized reversible PEM fuel cell. At low current density, the alkaline cell is obtaining round‐trip efficiencies close to 70%, while the PEM cell achieves only 60% efficiency. With Silver‐catalyst clear higher efficiencies up to 80% are presentable. The PEM fuel cell shows obvious better performance at high current densities.
The mediating role of sense of coherence on mental health outcomes in carers of older dependent relatives: A longitudinal study
Authors: López‐Martínez Catalina , Orgeta Vasiliki , Frías‐Osuna Antonio , del‐Pino‐Casado Rafael
Published: November 2020

Background

Sense of coherence (SOC) is an important protective factor for carer well‐being but research to date remains cross‐sectional, focusing primarily on the direct effects of SOC on carers' mental health. The study's aim was to investigate the mediating role of SOC in the longitudinal relationship between caregiver strain and carers' psychological health, and its stability over time.

Methods

Prospective longitudinal study conducted in Jaén (Spain) with a probabilistic sample of 132 carers of older people, with data collected at baseline and at 1‐year follow‐up. We measured SOC, caregiver strain, anxiety and depressive symptoms, and several care‐recipient characteristics and intensity of care provided. We used multiple linear regression modelling and the Sobel test to analyse mediation effects.

Results

SOC was significantly negatively longitudinally associated with both anxiety (β = −0.38, p = 0.001) and depressive symptoms (β = −0.28, p = 0.023), after controlling for several confounders. SOC mediated both the relationship between caregiver strain and anxiety, and caregiver strain and depressive symptoms (Sobel test: p < 0.001 for anxiety and p < 0.001 for depressive symptoms). Differences between baseline and 1‐year follow‐up SOC scores were not statistically significant (p = 0.617).

Conclusions

SOC appears to buffer the impact of caregiver strain on symptoms of depression and anxiety in informal carers of older people. Our data showed that SOC is an important psychological resource for carers that remained relatively stable under non‐experimental conditions over a period of 1 year in this sample. The findings suggest that interventions aimed at strengthening SOC may protect carer psychological well‐being.
Associations between cognitive function, actigraphy‐based and self‐reported sleep in older community‐dwelling adults: Findings from the Irish Longitudinal Study on Ageing
Authors: Scarlett Siobhan , Kenny Rose Anne , O'Connell Matthew DL , Nolan Hugh , de Looze Céline
Published: November 2020

Objective

Cognitive impairment is prevalent in older ages. Associations with sleep are well established; however, ambiguity remains in which sleep characteristics contribute to this impairment. We examined cross‐sectional associations between both self‐reported and actigraphy‐based sleep and cognitive performance across a number of domains in community‐dwelling older adults.

Methods

1520 participants aged 50 and older with self‐reported and actigraphy‐based total sleep time (TST) (≤5, 6, 7–8, 9 and ≥10 h) and self‐reported sleep problems were analysed. Cognitive function was assessed using the Mini‐Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), verbal fluency, immediate and delayed recall memory, colour trails tests, and choice reaction tests (CRT). Associations between sleep and cognition were modelled using linear and negative binomial regression.

Results

Negative associations were found between ≥10 h of self‐reported TST and MoCA error rate (incidence rate ratio [IRR] = 1.42; 95% confidence interval [CI] = 1.18, 1.71; p < 0.001); verbal fluency (beta [B] = −2.32 words; 95% CI = −4.00, −0.65; p < 0.01); and delayed recall (B = −0.91 words; 95% CI = −1.58, −0.25; p < 0.05) compared to 7–8 h. Significant associations with actigraphy‐based TST were limited to MoCA error rate in ≤5 h (IRR = 1.22; 95% CI = 1.02, 1.45; p < 0.05) compared to 7–8 h. Higher numbers of sleep problems were associated with slower performance in CRT cognitive response time (IRR = 1.02; 95% CI = 1.00, 104; p < 0.05) and total response time (IRR = 1.02; 95% CI = 1.00, 1.04; p < 0.05).

Conclusions

Self‐reported long sleep duration was consistently associated with worse cognitive performance across multiple domains. Marginal associations between cognition and both actigraphy‐based sleep and self‐reported sleep problems were also apparent. These results further affirm poor sleep as a risk factor for cognitive impairment.
Neutral Ceramidase Mediates Nonalcoholic Steatohepatitis by Regulating Monounsaturated Fatty Acids and Gut IgA+ B Cells
Authors: Gu Xuemei , Sun Rui , Chen Liang , Chu Shenghui , Doll Mark A. , Li Xiaohong , Feng Wenke , Siskind Leah , McClain Craig J. , Deng Zhongbin
Published: November 2020

Background and Aims

Nonalcoholic steatohepatitis (NASH) is associated with obesity and an increased risk for liver cirrhosis and cancer. Neutral ceramidase (NcDase), which is highly expressed in the intestinal brush border of the small intestine, plays a critical role in digesting dietary sphingolipids (ceramide) to regulate the balance of sphingosine and free fatty acids. It remains unresolved whether obesity‐associated alteration of NcDase contributes to the manifestation of NASH. Here, we revealed that NcDase deficiency in murine models of NASH prevents hepatic inflammation and fibrosis but not steatosis.

Approach and Results

NcDase−/− mice display reduced stearoyl‐CoA desaturase (SCD) 1 expression with a compositional decrease of monounsaturated fatty acids (MUFAs) under the different dietary conditions. We further found that NcDase is a functional regulator of intestinal B cells and influences the abundance and quality of the secretory IgA response toward commensal bacteria. Analysis of composition of the gut microbiota found that Clostridiales colonization was increased in NcDase−/− mice. The colonization of germ‐free mice with gut microbiota from NcDase−/− mice resulted in a greater decrease in the expression of SCD1 and the level of MUFAs in the liver relative to gut microbiota from wild‐type littermates, which are associated with the alternation of IgA‐bound bacteria, including increase of Ruminococcaceae and reduction of Desulfovibrio. Mechanistically, NcDase is a crucial link that controls the expression of SCD1 and MUFA‐mediated activation of the Wnt/β‐catenin. Very importantly, our experiments further demonstrated that Wnt3a stimulation can enhance the activity of NcDase in hepatocytes.

Conclusions

Thus, the NcDase‐SCD1‐Wnt feedback loop promotes the diet‐induced steatohepatitis and fibrosis through the regulation of intestinal IgA+ immune cells.
Persistent Control of Hepatitis B Virus and Hepatitis Delta Virus Infection Following REP 2139-Ca and Pegylated Interferon Therapy in Chronic Hepatitis B Virus/Hepatitis Delta Virus Coinfection
Authors: Bazinet Michel , Pântea Victor , Cebotarescu Valentin , Cojuhari Lilia , Jimbei Pavlina , Anderson Mark , Gersch Jeff , Holzmayer Vera , Elsner Carina , Krawczyk Adalbert , Kuhns Mary C. , Cloherty Gavin , Dittmer Ulf , Vaillant Andrew
Published: November 2020

Finite therapy of HBV/HDV co-infection with REP 2139-Ca and pegIFN has good long term safety and is accompanied by high rates of functional cure of HDV, normalization of liver function and additional functional cure of HBV. HBV functional cure is accompanied inactivation / clearance of cccDNA and clearance of integrated HBV DNA.

The nucleic acid polymer REP 2139 inhibits assembly/secretion of hepatitis B virus (HBV) subviral particles. Previously, REP 2139-Ca and pegylated interferon (pegIFN) in HBV/hepatitis delta virus (HDV) coinfection achieved high rates of HDV RNA and hepatitis B surface antigen (HBsAg) loss/seroconversion in the REP 301 study (NCT02233075). The REP 301-LTF study (NCT02876419) examined safety and efficacy during 3.5 years of follow-up. In the current study, participants completing therapy in the REP 301 study were followed for 3.5 years. Primary outcomes were safety and tolerability, and secondary outcomes were HDV functional cure (HDV RNA target not detected [TND], normal alanine aminotransferase [ALT]), HBV virologic control (HBV DNA ≤2,000 IU/mL, normal ALT), HBV functional cure (HBV DNA TND; HBsAg <0.05 IU/mL, normal ALT), and HBsAg seroconversion. Supplemental analysis included high-sensitivity HBsAg (Abbott ARCHITECT HBsAg NEXT), HBV pregenomic RNA (pgRNA), HBsAg/hepatitis B surface antibody (anti-HBs) immune complexes (HBsAg ICs), and hepatitis B core-related antigen (HBcrAg). Asymptomatic grade 1-2 ALT elevations occurred in 2 participants accompanying viral rebound; no other safety or tolerability issues were observed. During therapy and follow-up, HBsAg reductions to <0.05 IU/mL were also <0.005 IU/mL. HBsAg ICs declined in 7 of 11 participants during REP 2139-Ca monotherapy and in 10 of 11 participants during follow-up. HDV functional cure persisted in 7 of 11 participants; HBV virologic control persisted in 3 and functional cure (with HBsAg seroconversion) persisted in 4 of these participants. Functional cure of HBV was accompanied by HBV pgRNA TND and HBcrAg <lower limit of quantitation. Conclusion: REP 2139-Ca + pegIFN is not associated with long-term safety or tolerability issues. The establishment of HDV functional cure and HBV virologic control/functional cure and HBsAg seroconversion are durable over 3.5 years and may reflect removal of integrated HBV DNA from the liver. Further investigation is warranted in larger studies.
Effects of geological structures on rainfall‐runoff responses in headwater catchments in a sedimentary rock mountain
Authors: Inaoka Jun , Kosugi Ken'ichirou , Masaoka Naoya , Itokazu Tetsushi , Nakamura Kimihito
Published: November 2020

Clarifying rainfall‐runoff responses in mountainous areas is essential for disaster prediction as well as water resource management. Although runoff is considered to be significantly affected by topography, some previous studies have reported that geological structures also have significant effects on rainfall‐runoff characteristics. Particularly in headwater catchments located in sedimentary rock mountains, dips and strikes may significantly affect rainwater discharge. In this study, the effects of geological structures on rainfall‐runoff characteristics were investigated based on observed discharge hydrographs from 12 catchments, which lie radially from the summit of a sedimentary rock mountain. The results obtained were as follows: (1) Even though the topographic wetness index (TWI) distributions of the 12 catchments were similar, there were significant differences in their runoff characteristics; (2) Catchments with average flow direction oriented towards the strike direction (strike‐oriented catchments) are characterized by large baseflows; (3) Catchments with average flow direction oriented towards the opposite dip direction (opposite dip‐oriented catchments) are steep, and this results in quick storm runoff generation; (4) Catchments with average flow direction oriented toward the dip direction (dip‐oriented catchments) are gentle, and this results in delayed storm runoff generation. It was presumed that in strike‐oriented catchments, large quantities of groundwater flowing along the bedding planes owing to hydraulic anisotropy, exfiltrate and sustain the large amount of the observed baseflow, that is, in strike‐oriented catchments, runoff is directly controlled by geological structures. Conversely, in opposite dip‐oriented and dip‐oriented catchments, runoff is indirectly controlled by geological structures, that is, geological structures affect slope gradients, which result in differences in storm runoff generation. Thus, this study clearly illustrates that geological structures significantly affect rainfall‐runoff responses in headwater catchments located in sedimentary rock mountains.
An analytical solution for rapidly predicting post‐fire peak streamflow for small watersheds in southern California
Authors: Wilder Brenton A. , Lancaster Jeremy T. , Cafferata Peter H. , Coe Drew B. R. , Swanson Brian J. , Lindsay Donald N. , Short William R. , Kinoshita Alicia M.
Published: November 2020

Following wildfires, the probability of flooding and debris flows increase, posing risks to human lives, downstream communities, infrastructure, and ecosystems. In southern California (USA), the Rowe, Countryman, and Storey (RCS) 1949 methodology is an empirical method that is used to rapidly estimate post‐fire peak streamflow. We re‐evaluated the accuracy of RCS for 33 watersheds under current conditions. Pre‐fire peak streamflow prediction performance was low, where the average R2was 0.29 and average RMSE was 1.10 cms/km2for the 2‐ and 10‐year recurrence interval events, respectively. Post‐fire, RCS performance was also low, with an average R2of 0.26 and RMSE of 15.77 cms/km2for the 2‐ and 10‐year events. We demonstrated that RCS overgeneralizes watershed processes and does not adequately represent the spatial and temporal variability in systems affected by wildfire and extreme weather events and often underpredicted peak streamflow without sediment bulking factors. A novel application of machine learning was used to identify critical watershed characteristics including local physiography, land cover, geology, slope, aspect, rainfall intensity, and soil burn severity, resulting in two random forest models with 45 and five parameters (RF‐45 and RF‐5, respectively) to predict post‐fire peak streamflow. RF‐45 and RF‐5 performed better than the RCS method; however, they demonstrated the importance and reliance on data availability. The important parameters identified by the machine learning techniques were used to create a three‐dimensional polynomial function to calculate post‐fire peak streamflow in small catchments in southern California during the first year after fire (R2= 0.82; RMSE = 6.59 cms/km2) which can be used as an interim tool by post‐fire risk assessment teams. We conclude that a significant increase in data collection of high temporal and spatial resolution rainfall intensity, streamflow, and sediment loading in channels will help to guide future model development to quantify post‐fire flood risk.
Panel gene profiling of small bowel adenocarcinoma: Results from theNADEGEprospective cohort
Authors: Aparicio Thomas , Svrcek Magali , Henriques Julie , Afchain Pauline , Lièvre Astrid , Tougeron David , Gagniere Johan , Terrebonne Eric , Piessen Guillaume , Legoux Jean‐Louis , Lecaille Cédric , Pocard Marc , Gornet Jean‐Marc , Zaanan Aziz , Lavau‐Denes Sandrine , Lecomte Thierry , Deutsch David , Vernerey Dewi , Laurent Puig Pierre
Published: November 2020

Small bowel adenocarcinoma (SBA) is a rare tumour. Large genomic analyses with prognostic assessments are lacking. The NADEGE cohort has enrolled 347 patients with all stage SBA from 2009 to 2012. Next‐generation sequencing investigates the presence of 740 hotspot somatic mutations in a panel of 46 genes involved in carcinogenesis. The mismatch repair (MMR) status was assessed by immunochemistry. We have collected 196 tumour samples and 125 had conclusive results for mutation analysis. The number of mutations was 0 in 9.6% of tumours, only 1 in 32.0%, 2 in 26.4% and ≥3 in 32.0%. Overall, at least one genomic alteration was observed in 90.4% of tumour. The most frequent genomic alteration was inKRAS(44.0%),TP53(38.4%),PIK3CA(20.0%),APC(18.4%),SMAD4(14.4%) andERBB2(7.2%) genes.KRASmutations were more frequent in synchronous metastatic tumours than in localised tumours (72.7% vs 38.2%,P= .003). There was no significant difference in the mutation rates according to primary location for the most frequently altered gene.ATM,FGFR3andFGFR1gene alterations were associated with Lynch syndrome andIDH1mutations with Crohn disease. dMMR tumours were associated with younger age, localised tumours, lessKRASbut moreSMARCB1mutations. No genomic alteration was associated with overall survival. There is a trend for better survival in patient with dMMR tumours. In conclusion, there is a different genomic alteration profile in SBA according to predisposing diseases. No association between genomic alterations and prognoses was observed except for a trend of better prognoses associated with dMMR.
Risk of cardiovascular disease following gonadotropin‐releasing hormone agonists vs antagonists in prostate cancer: Real‐world evidence from five databases
Authors: George Gincy , Garmo Hans , Scailteux Lucie‐Marie , Balusson Frédéric , De Coster Greet , De Schutter Harlinde , Kuiper Josephina G. , Oger Emmanuel , Verbeeck Julie , Van Hemelrijck Mieke
Published: November 2020

Observational studies in prostate cancer (PCa) have shown an increased risk of cardiovascular disease (CVD) following gonadotropin‐releasing hormone (GnRH) agonists, whereas randomised‐controlled trials have shown no associations. Compared to GnRH agonists, GnRH antagonists have shown less atherosclerotic effects in preclinical models. We used real‐world data from five countries to investigate CVD risk following GnRH agonists and antagonists in PCa men. Data sources included cancer registries, primary and secondary healthcare databases. CVD event was defined as an incident or fatal CVD. Multivariable Cox proportional hazard models estimated hazard ratios (HRs) and 95% confidence intervals (CIs), which were pooled using random‐effects meta‐analysis. Stratified analyses were conducted by history of CVD and age (75 years). A total of 48 757 men were on GnRH agonists and 2144 on GnRH antagonists. There was no difference in risk of any CVD for men on GnRH antagonists and agonists (HR: 1.25; 95% CI: 0.96‐1.61; I2: 64%). Men on GnRH antagonists showed increased risk of acute myocardial infarction (HR: 1.62; 95% CI: 1.11‐2.35; I2: 0%) and arrhythmia (HR: 1.55; 95% CI: 1.11‐2.15, I2: 17%) compared to GnRH agonists. Having a history of CVD was found to be an effect modifier for the associations with some CVD subtypes. Overall, we did not observe a difference in risk of overall CVD when comparing GnRH antagonists with agonists—though for some subtypes of CVD we noted an increased risk with antagonists. Further studies are required to address potential confounding caused by unadjusted variables such as severity of CVD history and PCa stage.
Recovering the osteoblastic differentiation potential of mesenchymal stem cells derived from diabetic rats by photobiomodulation therapy
Authors: Bueno Natália Pieretti , Copete Isabella Nunes , Lopes Helena Bacha , Arany Praveen R. , Marques Márcia Martins , Ferraz Emanuela Prado
Published: November 2020

Autologous cell‐based therapy for bone regeneration might be impaired by diabetes mellitus (DM) due to the negative effects on mesenchymal stem cells (MSCs) differentiation. Strategies to recover their osteogenic potential could optimize the results. We aimed to evaluate the effect of photobiomodulation (PBM) therapy on osteoblast differentiation of rats with induced DM. Bone marrow MSCs of healthy and diabetic rats were isolated and differentiated into osteoblasts (OB and dOB, respectively). dOB were treated with PBM therapy every 72 hour (660 nm; 0.14 J; 20 mW; 0.714 W/cm2, and 5 J/cm2). Cell morphology, viability, gene and protein expression of osteoblastic markers, alkaline phosphatase (ALP) activity, and the mineralized matrix production of dOB‐PBM were compared to dOB. PBM therapy improved viability of dOB, increased the gene and protein expression of bone markers, the ALP activity and the mineralized matrix production. PBM therapy represents an innovative therapeutic approach to optimize the treatment of bone defects in diabetic patients.
Synthesis of coumarin derivatives in a microfluidic flow system employing the Pechmann condensation: A case study
Authors: Liu Ling‐Kang , Deng Jhao‐Hong , Guo Yang‐Ming
Published: November 2020

For the synthesis of coumarin derivatives using the Pechmann condensation scheme, an acidic ionic liquid catalyst, abbreviated as [EBsImH][HSO4], was prepared from the ring opening of 1,4‐butanesultone by 1‐ethylimidazole, followed by the addition of 1 equiv. H2SO4(c). The [EBsImH][HSO4]‐catalyzed Pechmann condensation reactions proceeded smoothly in a batch setup, with recyclable [EBsImH][HSO4] showing great catalytic activity. The acidic ionic liquid catalyst [EBsImH][HSO4] was recovered from EtOAc/H2O extraction of the product mixture, where the H2O layer was worked up and dried for reuse in consecutive runs of the Pechmann condensation reactions, maintaining >85% conversion for four times. The catalytic reactions were also carried out in a microfluidic flow setup. The flow parameters, the reactant molar amounts, and the additional H2SO4 as a modifying acid catalyst were optimized in the current case study. A minimum conversion rate of 2.8 g/hr of coumarin derivatives was demonstrated.
Electrochemical detection of fucosidase activity in human serum
Authors: Tsai Hweiyan , Yu Chen Yu , Chen Chien‐Hong , Fuh Chwan Bor
Published: November 2020

Liver cancer is one of the most common malignant tumors globally and is the second leading cause of cancer deaths in Taiwan. The activity of α‐fucosidase (AFU) in the human body has been confirmed by many clinical studies and experiments to be applicable in the early diagnosis of primary liver cancer. Herein, an electrochemical method for determining AFU activity was developed based on measuring p‐nitrophenol (pNP), which is a product of the reaction between AFU and p‐nitrophenyl‐α‐L‐fucopyranoside. The electrochemical detection of pNP in serum samples exhibited a wide linear range (2.5–250 μM), which corresponds to approximately 2.5–250 U/L AFU. The recovery of AFU measured upon enzyme addition to standard serum was 97–110%. Moreover, no potential interferants had a significant effect on pNP quantification. In contrast, an optical absorption method was subject to interference by the serum color. Further, the linear range for pNP was only 2.5–50 μM and the recovery of AFU was 165–219%. The performance of this absorption method could lead to a misjudgment of the disease course. Therefore, the electrochemical method for AFU activity determination has greater potential for use in clinical applications.
Clinical significance of the serum IgM and IgG to SARS‐CoV‐2 in coronavirus disease‐2019
Authors: Wu Li‐xiang , Wang Hui , Gou Dan , Fu Gang , Wang Jing , Guo Bian‐qin
Published: November 2020

Objective

To explore the clinical value of serum IgM and IgG to SARS‐CoV‐2 in COVID‐19.

Methods

105 COVID‐19 patients were enrolled as the disease group. 197 non‐COVID‐19 patients served as the control group. Magnetic chemiluminescent immunoassay (MCLIA) was used to detect the IgM and IgG.

Results

The peak of positive rates of SARS‐CoV‐2 IgM was about 1 week earlier than that of IgG. It reached to peak within 15–21 days and then began a slowly decline. The positive rates of IgG were increased with the disease course and reached the peak between 22 and 39 days. The differences in sensitivity of the three detection modes (IgM, IgG, and IgM + IgG) were statistically significant. The largest group of test cases (illness onset 15–21 days) showed that the positive rate of IgG was higher than IgM. Also, the sensitivity of IgM combined with IgG was higher than IgM or IgG. IgM and IgG were monitored dynamically for 16 patients with COVID‐19, the results showed that serological transformation of IgM was carried out simultaneously with IgG in seven patients, which was earlier than IgG in four patients and later than IgG in five patients.

Conclusion

The detection of SARS‐CoV‐2 IgM and IgG is very important to determine the course of COVID‐19. Nucleic acid detection combined with serum antibody of SARS‐CoV‐2 may be the best laboratory indicator for the diagnosis of SARS‐CoV‐2 infection and the phrase and predication for prognosis of COVID‐19.
Morphine promotes tumorigenesis and cetuximab resistance via EGFR signaling activation in human colorectal cancer
Authors: Lu Hong , Zhang Hao , Weng Mei‐lin , Zhang Jin , Jiang Nan , Cata Juan P. , Ma Duan , Chen Wan‐Kun , Miao Chang‐Hong
Published: November 2020

Morphine, a mu‐opioid receptor (MOR) agonist, has been extensively used to treat advanced cancer pain. In particular, in patients with cancer metastasis, both morphine and anticancer drugs are given simultaneously. However, evidence showed that morphine might be a risk factor in promoting the tumor's malignant potential. In this study, we report that treatment with morphine could activate MOR and lead to the promotion of proliferation, migration, and invasion in HCT116 and DLD1 colorectal cancer (CRC) cells with time‐concentration dependence. Moreover, morphine can also contribute to cetuximab's drug resistance, a targeted drug widely used to treat advanced CRC by inducing the activation of epidermal growth factor receptor (EGFR). The cell phenotype includes proliferation, migration, invasion, and drug resistance, which may be reversed by MOR knockdown or adding nalmefene, the MOR receptor antagonist. Receptor tyrosine kinase array analysis revealed that morphine selectively induced the transactivation of EGFR. EGFR transactivation resulted in the activation of ERK1/2 and AKT. In conclusion, morphine induces the transactivation of EGFR via MOR. It activates the downstream signal pathway AKT‐MTOR and RAS‐MAPK, increases proliferation, migration, and invasion, and promotes resistance to EGFR inhibitors in a CRC cell line. Furthermore, we verified that EGFR inhibition by cetuximab strongly reversed the protumoral effects of morphine in vitro and in vivo. Collectively, we provide evidence that morphine‐EGFR signaling might be a promising therapeutic target for CRC patients, especially for cetuximab‐resistant CRC patients.