bluuebunny/crossref_metadata_2025_split huggingface dataset:
Improved fluorescence assays to measure the defects associated with F508del‐CFTR allow identification of new active compounds
Authors: Langron Emily , Simone Michela I , Delalande Clémence M S , Reymond Jean‐Louis , Selwood David L , Vergani Paola 
Published: January 2017

Background and PurposeCystic fibrosis (CF) is a debilitating disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which codes for a Clˉ/HCO3ˉ channel. F508del, the most common CF‐associated mutation, causes both gating and biogenesis defects in the CFTR protein. This paper describes the optimization of two fluorescence assays, capable of measuring CFTR function and cellular localization, and their use in a pilot drug screen.Experimental ApproachHEK293 cells expressing YFP‐F508del‐CFTR, in which halide sensitive YFP is tagged to the N‐terminal of CFTR, were used to screen a small library of compounds based on the VX‐770 scaffold. Cells expressing F508del‐CFTR‐pHTomato, in which a pH sensor is tagged to the fourth extracellular loop of CFTR, were used to measure CFTR plasma membrane exposure following chronic treatment with the novel potentiators.Key ResultsActive compounds with efficacy ~50% of VX‐770, micromolar potency, and structurally distinct from VX‐770 were identified in the screen. The F508del‐CFTR‐pHTomato assay suggests that the hit compound MS131A, unlike VX‐770, does not decrease membrane exposure of F508del‐CFTR.Conclusions and ImplicationsMost known potentiators have a negative influence on F508del‐CFTR biogenesis/stability, which means membrane exposure needs to be monitored early during the development of drugs targeting CFTR. The combined use of the two fluorescence assays described here provides a useful tool for the identification of improved potentiators and correctors. The assays could also prove useful for basic scientific investigations on F508del‐CFTR, and other CF‐causing mutations.
Ribosomal protein S3 gene silencing protects against experimental allergic asthma
Authors: Dong Jinrui , Liao Wupeng , Peh Hong Yong , Chan Tze Khee , Tan W S Daniel , Li Li , Yong Amy , Wong W S Fred 
Published: January 2017

Background and PurposeRibosomal protein S3 (RPS3) is a 40S ribosomal protein of the S3P family essential for implementing protein translation. RPS3 has recently been found to interact with the p65 subunit of the NF‐κB complex and promote p65 DNA‐binding activity. Persistent activation of the NF‐κB pathway is evident in allergic asthma. We hypothesized that gene silencing of lung RPS3 can ameliorate allergic airway inflammation.Experimental ApproachThe gene silencing efficacy of RPS3 siRNA was screened in three different mouse cell lines by real‐time PCR and immunoblotting. Protective effects of intratracheal RPS3 siRNA in a house dust mite (HDM) mouse asthma model were determined by measuring cell counts in lung lavage fluid and lung sections, lung cytokine profiles and airway hyperresponsiveness (AHR).Key ResultsRPS3 siRNA markedly knocked down RPS3 levels in all mouse cell lines tested, and in mouse lung tissues, blocked TNF‐α‐ or HDM‐induced release of mediators by the cultured cells and reduced eosinophil counts in lung lavage fluid from the HDM mouse asthma model. RPS3 siRNA lessened HDM‐induced airway mucus hypersecretion, cytokine production and serum IgE elevation. Moreover, RPS3 knockdown significantly suppressed methacholine‐induced AHR in experimental asthma. RPS3 siRNA disrupted TNF‐α‐induced NF‐κB activation in a NF‐κB reporter gene assay in vitro and prevented the nuclear accumulation of p65 subunit and p65 transcriptional activation in HDM‐challenged lungs and cells.Conclusions and ImplicationsRPS3 gene silencing ameliorates experimental asthma, probably by disrupting NF‐κB activity. RPS3 could be a novel therapeutic target for allergic airway inflammation.
Endothelin‐1 and its receptors on haemorrhoidal tissue: a potential site for therapeutic intervention
Authors: Lohsiriwat Varut , Scholefield John H , Wilson Vincent G , Dashwood Michael R 
Published: January 2017

Background and PurposeHaemorrhoids is a common anorectal condition affecting millions worldwide. We have studied the effect of endothelin‐1 (ET‐1) and the role of endothelin ETA and ETB receptors in haemorrhoid tissue.Experimental ApproachProtein expression of ET‐1, ETA and ETB receptors were compared between haemorrhoids and normal rectal submucosa using Western blot analysis, with the localization of proteins determined by autoradiography and immunohistochemistry. Effects of ET‐1 and sarafotoxin 6a on human colonic and rectal arteries and veins was assessed by wire myography and the involvement of receptor subtypes established by selective antagonists.Key ResultsDense binding of [125I]‐ET‐1 to haemorrhoidal sections was reduced by selective receptor antagonists. A higher density of ETB than ETA receptors was found in haemorrhoidal, than in control rectal tissue and confirmed by Western blot analysis. ETA and ETB receptors were localized to smooth muscle of haemorrhoidal arteries and veins, with ETB receptors on the endothelium. Human colonic and rectal arteries and veins were similarly sensitive to ET‐1 and affected by the ETA selective antagonist, but sarafotoxin S6a‐induced contractions were more pronounced in veins and antagonized by a selective ETB receptor antagonist.Conclusions and ImplicationsETA and ETB receptors are present in human haemorrhoids with ETB receptors predominating. ETA receptors are activated by ET‐1 to mediate a contraction in arteries and veins, but the latter are selectively activated by sarafotoxin S6a – a response that involves ETB receptors at low concentrations. Selective ETB agonists may have therapeutic potential to reduce congestion of the haemorrhoidal venous sinusoids.
Monolithic implant‐supported lithium disilicate (LS2) crowns in a complete digital workflow: A prospective clinical trial with a 2‐year follow‐up
Authors: Joda Tim , Ferrari Marco , Brägger Urs 
Published: January 2017

AbstractBackgroundThe technical development of digital processing allows the production of anatomically full‐contoured implant‐supported restorations.PurposeThe aim of this prospective clinical trial was to analyze the treatment concept of monolithic lithium disilicate (LS2) single‐unit restorations in a complete digital workflow.Material and MethodsForty‐four patients were restored with 50 screw‐retained monolithic implant LS2 crowns bonded to pre‐fabricated titanium abutments on soft tissue level implants (Institut Straumann AG, Basel, Switzerland) in premolar and molar sites. All implant restorations were digitally designed after intraoral optical scanning (IOS) and CAD/CAM‐processing without physical model situations. Study participants were clinically and radiographically examined based on an annually performed follow‐up. The “Functional Implant Prosthodontic Score” (FIPS) was applied for objective outcome assessment after 2 years of loading. Five variables were defined for FIPS evaluation, resulting in a maximum score of 10 per implant restoration. Descriptive statistics were calculated for mean scores standard deviations, medians, and Q25–Q75.ResultsAll patients could be successfully treated within two clinical appointments. No clinical modifications were necessary for the seating of the monolithic crowns, neither for interproximal nor occlusal sites. The implant LS2 restorations demonstrated survival rates of 100% without any technical or biological complications after 2 years. The mean total FIPS score was 7.7 ± 1.0, ranging from 6 to 10.ConclusionsCAD/CAM‐produced monolithic implant crowns out of LS2 in a complete digital workflow seem to be a feasible treatment concept for the rehabilitation of single‐tooth gaps in posterior sites under mid‐term observation.
Quantifying mammal biodiversity co‐benefits in certified tropical forests
Authors: Sollmann Rahel , Mohamed Azlan , Niedballa Jürgen , Bender Johannes , Ambu Laurentius , Lagan Peter , Mannan Sam , Ong Robert C. , Langner Andreas , Gardner Beth , Wilting Andreas 
Published: January 2017

AbstractAimFinancial incentives to manage forests sustainably, such as certification or carbon storage payments, are assumed to have co‐benefits for biodiversity conservation. This claim remains little studied for rain forest mammals, which are particularly threatened, but challenging to survey.LocationSabah, Malaysia, Borneo.MethodsWe used photographic data from three commercial forest reserves to show how community occupancy modelling can be used to quantify mammalian diversity conservation co‐benefits of forest certification. These reserves had different management histories, and one was certified by the Forest Stewardship Council.ResultsMany threatened species occupied larger areas in the certified reserve. Species richness, estimated per 200 × 200‐m grid cell throughout all reserves, was higher in the certified site, particularly for threatened species. The certified reserve held the highest aboveground biomass. Within reserves, aboveground biomass was not strongly correlated with patterns of mammal richness (Spearman's rho from 0.03 to 0.32); discrepancies were strongest along reserve borders.Main conclusionsOur approach provides a flexible and standardized tool to assess biodiversity and identify winners of sustainable forestry. Inferring patterns of species richness from camera‐trapping carries potential for the objective designation of high conservation value forest. Correlating species richness with aboveground biomass further allows evaluating the biodiversity co‐benefits of carbon protection. These advantages make the present approach an ideal tool to overcome the difficulties to rigorously quantify biodiversity co‐benefits of forest certification and carbon storage payments.
Effect of valsartan on kidney outcomes in people with impaired glucose tolerance
Authors: Currie Gemma , Bethel M. Angelyn , Holzhauer Björn , Haffner Steven M. , Holman Rury R. , McMurray John J. V. 
Published: January 2017

AimsTo examine the effect of valsartan on kidney outcomes in patients with impaired glucose tolerance (IGT).MethodsIn a double‐blind randomized trial, 9306 patients with IGT were assigned to valsartan (160 mg daily) or placebo. The co‐primary endpoints were the development of diabetes and two composite cardiovascular outcomes. Prespecified renal endpoints included: the composite of renal death, end‐stage renal disease (ESRD) or doubling of serum creatinine; estimated glomerular filtration rate (eGFR) ≤30 mL/min/1.73 m2; hospitalization for renal failure; and progression from normoalbuminuria to microalbuminuria, microalbuminuria to macroalbuminuria, and normoalbuminuria to macroalbuminuria. The median follow‐up was 6.2 years.ResultsValsartan reduced the incidence of diabetes but not cardiovascular events. In the valsartan group, 25/4631 patients (0.5%), vs 26/4675 (0.6%) patients in the placebo group, developed ESRD or experienced doubling of serum creatinine (hazard ratio [HR] 0.96, 95% confidence interval [CI] 0.55‐1.66;
P
= .87). Few patients in either group developed an eGFR of ≤30 mL/min/1.73 m2 or had a renal hospitalization. Fewer patients on valsartan (237/4084 [5.8%]) than on placebo (342/4092 [8.4%]) developed microalbuminuria (HR 0.68, 95% CI 0.57‐0.80;
P
< .0001), and fewer valsartan‐treated patients developed macroalbuminuria. Overall, urinary albumin‐to‐creatinine ratio (UACR) was 11% lower with valsartan (95% CI 8‐13;
P
< .0001) and 9% lower (95% CI 6‐11;
P
< .0001) after adjusting for both glucose and blood pressure.ConclusionsThe effect of valsartan on UACR was not wholly explained by change in blood pressure or glucose. Valsartan reduced the incidence of microalbuminuria in IGT without increasing the incidence of hyperkalaemia or renal dysfunction compared with placebo.
Immunohistochemical assessment of glucagon‐like peptide 1 receptor (GLP‐1R) expression in the pancreas of patients with type 2 diabetes
Authors: Kirk Rikke K. , Pyke Charles , von Herrath Matthias G. , Hasselby Jane P. , Pedersen Lars , Mortensen Pia G. , Knudsen Lotte B. , Coppieters Ken 
Published: January 2017

AimsGlucagon‐like peptide‐1 (GLP‐1) is an incretin hormone which stimulates insulin release and inhibits glucagon secretion from the pancreas in a glucose‐dependent manner. Incretin‐based therapies, consisting of GLP‐1 receptor (GLP‐1R) agonists and dipeptidyl peptidase‐4 (DPP‐4) inhibitors, are used for the treatment of type 2 diabetes (T2D). Immunohistochemical studies for GLP‐1R expression have been hampered previously by the use of unspecific polyclonal antibodies. This study aimed to assess the expression levels of GLP‐1R in a set of T2D donor samples obtained via nPOD.MethodsThis study used a new monoclonal antibody to assess GLP‐1R expression in pancreatic tissue from 23 patients with T2D, including 7 with a DPP‐4 inhibitor and 1 with a history of GLP‐1R agonist treatment. A software‐based automated image analysis algorithm was used for quantitating intensities and area fractions of GLP‐1R positive compartments.ResultsThe highest intensity GLP‐1R immunostaining was seen in beta‐cells in islets (average signal intensity, 76.1 [±8.1]). GLP‐1R/insulin double‐labelled single cells or small clusters of cells were also frequently located within or in close vicinity of ductal epithelium in all samples and with the same GLP‐1R immunostaining intensity as found in beta‐cells in islets. In the exocrine pancreas a large proportion of acinar cells expressed GLP‐1R with a 3‐fold lower intensity of immunoreactivity as compared to beta‐cells (average signal intensity 25.5 [±3,3]). Our studies did not unequivocally demonstrate GLP‐1R immunoreactivity on normal‐appearing ductal epithelium. Pancreatic intraepithelial neoplasia (PanINs; a form of non‐invasive pancreatic ductular neoplasia) was seen in most samples, and a minority of these expressed low levels of GLP‐1R.ConclusionThese data confirm the ubiquity of early stage PanIN lesions in patients with T2D and do not support the hypothesis that incretin‐based therapies are associated with progression towards the more advanced stage PanIN lesions.
A Best Practice Modular Design of a Hybrid Course Delivery Structure for an Executive Education Program
Authors: Klotz Dorothy E. , Wright Thomas A. 
Published: January 2017

ABSTRACTThis article highlights a best practice approach that showcases the highly successful deployment of a hybrid course delivery structure for an Operations core course in an Executive MBA Program. A key design element of the approach was the modular design of both the course itself and the learning materials. While other hybrid deployments may stress the importance of modular design, our deployment demonstrates how content can be easily mixed and matched into shorter courses on select topics. In particular, through the migration of select content online and the use of face‐to‐face class time for discussion and active‐learning experiences, we were able to dramatically reduce face‐to‐face student seat time, positively increase the student experience, and facilitate dramatic improvement in student academic achievement.
Incipient speciation in theAnastrepha fraterculuscryptic species complex: reproductive compatibility betweenA. sp.1 aff.fraterculusandA. sp.3 aff.fraterculus
Authors: Roriz Alzira Kelly Passos , Japyassú Hilton F. , Joachim‐Bravo Iara Sordi 
Published: January 2017

AbstractReproductive isolation among biological strains can be detected by analyzing reproductive barriers between populations. TheAnastrepha fraterculusWiedemann (Diptera:Tephritidae) cryptic species complex comprises eight morphotypes defined by morphometric characteristics. The present study analyzed the existence of pre‐ and post‐zygotic isolation among populations ofA. sp.1 aff.fraterculus(A. sp.1) andA. sp.3 aff.fraterculus(A. sp.3) determined by tests of sexual compatibility, analyses of the temporal pattern of calling behavior, and the reproductive performance of the offspring of homotypic and heterotypic crosses. Pre‐zygotic tests indicated asymmetric matings with preferences for homotypic crosses. TheA. sp.3 population demonstrated distinct characteristics, including low copulation percentages and a high proportion of homotypic copulations under crowded conditions. The females of theA. sp.1 population demonstrated lower copulation latency. The analyzed morphotypes demonstrated two peaks in pheromone emission activity, at the beginning and at the end of the morning and were differentiated in the abundance of males. The calling behavior of the hybrids betweenA. sp.1 male andA. sp.3 female crosses had an intermediate pattern compared to either parental type. Post‐zygotic compatibility tests indicated low viability of the cross betweenA. sp.3 males andA. sp.1 females. Pronounced asymmetries were found in the sex ratios of the offspring of heterotypic crosses, and only hybrids fromA. sp.1 male vs.A. sp.3 female crosses produced descendants. The observed pre‐ and post‐zygotic incompatibilities reflect the probable occurrence of incipient speciation betweenA. sp.1 andA. sp.3. A more adequate taxonomic classification of this species complex that considers the distinct characteristics of each morphotype will be needed to improve environmentally wise control methods against this insect pest.
Rapid recovery of a population of the cryptic and evolutionarily distinct Hochstetter's Frog, Leiopelma hochstetteri, in a pest‐free environment
Authors: Longson Christopher G. , Brejaart Ria , Baber Matthew J. , Babbitt Kimberly J. 
Published: January 2017

SummaryNew Zealand's native frogs (genus Leiopelma) display a very high degree of endemism and belong to a distinctive and ancient evolutionary lineage. All four extant species are considered to be threatened or endangered, but the mechanisms behind their decline are poorly understood. Many of the potential factors causing population declines are confounded for the two species living on the mainland; in particular, habitat degradation and the presence of introduced mammalian pests have been difficult to disentangle in the past. Here we present data on a population of Hochstetter's Frog (Leiopelma hochstetteri) living in a pest‐free mainland sanctuary, the 3363‐ha Maungatautari Ecological Island. Survey data from this population shows a fourfold increase in numbers between 2009 and 2012 from a very low initial density. Our results suggest that for Hochstetter's Frog at least, and possibly the other mainland species of Leiopelma, introduced mammals are a major driving factor in population declines. Given that the frog population at Maungatautari was discovered by serendipity when planning for the pest‐free sanctuary was already well underway, this also highlights the conservation value of large pest‐free areas and ecosystem‐level management.
Conserving and restoring endangered southern populations of the Squirrel Glider (Petaurus norfolcensis) in agricultural landscapes
Authors: Crane Mason , Lindenmayer David B. , Banks Sam C. 
Published: January 2017

SummarySouthern Squirrel Glider (Petaurus norfolcensis) populations are genetically distinct and generally found in the agricultural landscapes inland of Australia's Great Dividing Range. These populations are considered to be under greater threat of extinction than northern, coastal populations and face a unique set of environmental conditions and conservation challenges. For these reasons, we suggest that southern populations qualify as a separate evolutionarily significant unit to those from the northern, coastal segment of the range and should be managed separately. We summarize the species’ ecology specific to southern populations and relevant to management. We conduct a basic SWOT (strengths, weaknesses, opportunities, threats) analysis to highlight potential future management directions. From our review of new and existing ecological data and SWOT analysis, we outline ten points of action important for securing the future of southern Squirrel Glider populations.
Granzyme A potentiates chemokine production in IL‐17‐stimulated keratinocytes
Authors: Cheuk Stanley , Martini Elisa , Bergh Kerstin , Chang David , Rethi Bence , Ståhle Mona , Eidsmo Liv 
Published: January 2017

AbstractPlaque psoriasis presents with focal skin inflammation, partially maintained by IL‐17‐mediated interactions between infiltrating epidermal T cells and activated keratinocytes. Here we show that the majority of lesional epidermal CD8 T cells express granzyme A, alone or in combination with IL‐17. To assess proinflammatory properties of granzyme A in psoriasis, primary human keratinocytes were stimulated with granzyme A in the presence or absence of IL‐17. Out of 33 analysed keratinocyte‐derived inflammatory mediators, granzyme A potentiated IL‐17‐induced secretion of CXCL 1, CXCL 12 and CCL 4. Intriguingly, all three chemokines are implicated in psoriasis pathogenesis and are involved in recruitment of T cells, neutrophils and pDCs into inflamed tissues. Our results indicate that granzyme A produced by lesional CD8 T cells specifically increase the chemokine production from inflamed keratinocytes, thereby amplifying a chemotactic inflammatory loop that sustains psoriasis lesions.
Divergence of cAMP signalling pathways mediating augmented nucleotide excision repair and pigment induction in melanocytes
Authors: Wolf Horrell Erin M. , Jarrett Stuart G. , Carter Katharine M. , D'Orazio John A. 
Published: January 2017

AbstractLoss‐of‐function melanocortin 1 receptor (MC1R) polymorphisms are common in UV‐sensitive fair‐skinned individuals and are associated with blunted cAMP second messenger signalling and higher lifetime risk of melanoma because of diminished ability of melanocytes to cope with UV damage. cAMP signalling positions melanocytes to resist UV injury by upregulating synthesis of UV‐blocking eumelanin pigment and by enhancing the repair of UV‐induced DNA damage. cAMP enhances melanocyte nucleotide excision repair (NER), the genome maintenance pathway responsible for the removal of mutagenic UV photolesions, through cAMP‐activated protein kinase (protein kinase A)‐mediated phosphorylation of the ataxia telangiectasia‐mutated and Rad3‐related (ATR) protein on the S435 residue. We investigated the interdependence of cAMP‐mediated melanin upregulation and cAMP‐enhanced DNA repair in primary human melanocytes and a melanoma cell line. We observed that the ATR‐dependent molecular pathway linking cAMP signalling to the NER pathway is independent of MITF activation. Similarly, cAMP‐mediated upregulation of pigment synthesis is independent of ATR, suggesting that the key molecular events driving MC1R‐mediated enhancement of genome maintenance (eg PKA‐mediated phosphorylation of ATR) and MC1R‐induced pigment induction (eg MITF activation) are distinct.