bluuebunny/crossref_metadata_2025 huggingface dataset:
Proactive use of eltrombopag before the onset of clinical bleeding in two children with immune thrombocytopenia and lifestyle restrictions
Authors: Bergmann Shayla
Published: September 2017

Key Clinical Message

Children with immune thrombocytopenia (ITP) are often managed using a watch‐and‐wait approach to avoid conventional treatment that may be poorly tolerated. However, in some patients, this approach may lead to lifestyle restrictions due to risk of injury‐related bleeding. Eltrombopag is a well‐tolerated new option that may help these children.
A rare case of CD1a‐negative Langerhans cell histiocytosis of the central nervous system in a child
Authors: Powell Priscilla , Vitug Gaile , Castro‐Silva Fernando , Ray Anish
Published: September 2017

Key Clinical Message

Langerhans cell histiocytosis is a dendritic cell disorder with a wide spectrum of severity and presentations. Histopathology typically demonstrates a proliferation of Langerhans cells and a lymphohistiocytic inflammatory infiltrate with eosinophils. The diagnosis is supported by immunohistochemistry with the cell markers S100, CD1a, CD68, and Langerin [Blood, 126, 2015, 26 and N Engl J Med, 331, 1994, 154].
On the Redox Reactivity of a Geometrically Constrained Phosphorus(III) Compound
Authors: Robinson Thomas P. , De Rosa Daniel , Aldridge Simon , Goicoechea Jose M.
Published: September 2017

The reactivity of a geometrically constrained phosphorus(III) complex bearing the N,N‐bis(3,5‐di‐tert‐butyl‐2‐phenolate)amide pincer ligand (P(ONO); 1) towards oxidants and reductants is explored. This compound can be readily oxidized to the phosphorus(V) dihalo‐derivatives P(ONO)X2 where X=Cl (2), Br (3) and I (4). Attempts at isolating the analogous difluoride through oxidation of 1 were unsuccessful yielding only the hydrofluoride P(ONO)(H)F (5), however P(ONO)F2 (6) can be accessed via a halide exchange reaction of 2 with KF. Compound 2 can be employed as a precursor to novel cationic species through chloride ion displacement using strong Lewis bases. Thus, reaction of 2 with two or three molar equivalents of dimethylaminopyridine (DMAP) affords [P(ONO)(Cl)(DMAP)2]+ (7) and [P(ONO)(DMAP)3]2+ (8). Reaction of 2 with the weaker bidentate base 2,2′‐bipyridine (bipy) affords [P(ONO)(Cl)(bipy)]+ (9), although this species was only accessible upon addition of a halide abstracting agent. The dicationic tris(pyridine) adduct [P(ONO)(py)3]2+ (10) is also accessible by reaction of 4 with pyridine. Oxidation of 1 using oxygen gas proceeds slowly and allows for the observation of two compounds, a mixed valence dimeric phosphorus(III)/phosphorus(V) compound [P(ONO)(μ2‐O)(μ2:κ1,κ2‐ONO)P] (11) and the fully oxidized species [P(ONO)(μ2‐O)(μ2:κ1,κ2‐ONO)P(O)] (12). Finally, reaction of 1 using KC8 results in the dimerization of the putative radical anion [P(ONO)].− through formation of a P−P bond to afford [P(ONO)]22− (13). Reactions with TEMPO result in the formation of the trigonal bipyramidal species P(ONO)(TEMPO)2 (14).
Seasonal variation in hospitalization outcomes in patients admitted for heart failure in the United States
Authors: Akintoye Emmanuel , Briasoulis Alexandros , Egbe Alexander , Adegbala Oluwole , Alliu Samson , Sheikh Muhammad , Singh Manmohan , Ahmed Abdelrahman , Mallikethi‐Reddy Sagar , Levine Diane
Published: September 2017

Background

There is lack of evidence of the impact of varying season on heart failure (HF) hospitalization outcomes in the U.S.

Hypothesis

HF hospitalization outcomes exhibit significant seasonal variation in the U.S.

Methods

Using data from the National Inpatient Sample (2011‐2013), seasonal variation was classified based on meteorological classification of Northern Hemisphere–Spring, Summer, Fall, & Winter–and analysis was conducted via multivariable‐adjusted mixed‐effect model.

Results

An estimated 2.8 million adults were hospitalized for HF in the U.S. from 2011 to 2013. Of all hospitalizations, admissions were highest in Winter (27%), followed by Spring (26%), Fall (24%), and Summer (23%). The overall mortality rate was 3.1%. Compared with Spring, there was significantly lower mortality in Summer (odds ratio [OR]: 0.95, 95% CI: 0.91‐0.99) and Fall (OR: 0.94, 95% CI: 0.89‐0.98), but the highest mortality was in Winter (OR: 1.06, 95% CI: 1.02‐1.11). In addition, mean length of stay and median cost of hospitalization were highest in Winter (5.3 days, USD7459, respectively) and lowest in Summer (5.1 days, USD7181, respectively). However, age and sex differences existed (e.g. seasonal variation in inpatient mortality was only significant for patients age ≥65 years, and, compared

with the Spring season, males had higher risk of inpatient mortality in Winter (OR: 1.10, 95% CI: 1.04‐1.17) and females had lower risk of inpatient mortality in Summer (OR: 0.94, 95% CI: 0.88‐1.00) and Fall (OR: 0.92, 95% CI: 0.87‐0.98).

Conclusions

Among HF patients in the U.S., hospitalization outcomes were worse in Winter but better in Summer.
Sulfocoumarin‐, Coumarin‐, 4‐Sulfamoylphenyl‐Bearing Indazole‐3‐carboxamide Hybrids: Synthesis and Selective Inhibition of Tumor‐Associated Carbonic Anhydrase Isozymes IX and XII
Authors: Angapelly Srinivas , Sri Ramya P. V. , Angeli Andrea , Supuran Claudiu T. , Arifuddin Mohammed
Published: September 2017

A series of sulfocoumarin‐, coumarin‐, and 4‐sulfamoylphenyl‐bearing indazole‐3‐carboxamide hybrids were synthesized and investigated as inhibitors of the human carbonic anhydrase (hCA, EC 4.2.1.1) isoforms I and II (cytosolic isozymes), as well as hCA IX and XII (transmembrane, tumor‐associated enzymes). Compounds 6 a–g (amide derivatives) and 7 a–h (triazoles) act as “prodrugs”, and their hydrolysis products are the de facto CA inhibitors. These compounds displayed sub‐micromolar to high‐nanomolar inhibitory activity against hCA isoforms IX and XII, which were recently validated as antitumor drug targets. Moreover, no inhibition of the off‐target hCA I and II isoforms was observed. Compounds 8 a–f (another set of triazoles) exhibited nanomolar inhibition against hCA isoforms I, II, IX and XII, among which compounds 8 c, 8 d, and 8 f were found to inhibit the tumor‐associated hypoxia‐induced hCA isoform IX with Ki values of 1.8, 2.3, and 2.0 nm respectively. Further exploration of these compounds could be useful for the development of novel antitumor agents with selective mechanisms of CA inhibitory action.
Lectin‐Array Blotting
Authors: Pazos Raquel , Echevarria Juan , Hernandez Alvaro , Reichardt Niels‐Christian
Published: September 2017

Aberrant protein glycosylation is a hallmark of cancer, infectious diseases, and autoimmune or neurodegenerative disorders. Unlocking the potential of glycans as disease markers will require rapid and unbiased glycoproteomics methods for glycan biomarker discovery. The present method is a facile and rapid protocol for qualitative analysis of protein glycosylation in complex biological mixtures. While traditional lectin arrays only provide an average signal for the glycans in the mixture, which is usually dominated by the most abundant proteins, our method provides individual lectin binding profiles for all proteins separated in the gel electrophoresis step. Proteins do not have to be excised from the gel for subsequent analysis via the lectin array but are transferred by contact diffusion from the gel to a glass slide presenting multiple copies of printed lectin arrays. Fluorescently marked glycoproteins are trapped by the printed lectins via specific carbohydrate‐lectin interactions and after a washing step their binding profile with up to 20 lectin probes is analyzed with a fluorescent scanner. The method produces the equivalent of 20 lectin blots in a single experiment, giving detailed insight into the binding epitopes present in the fractionated proteins. © 2017 by John Wiley & Sons, Inc.
Automated Tracking of Cell Migration with Rapid Data Analysis
Authors: DuChez Brian J.
Published: September 2017

Cell migration is essential for many biological processes including development, wound healing, and metastasis. However, studying cell migration often requires the time‐consuming and labor‐intensive task of manually tracking cells. To accelerate the task of obtaining coordinate positions of migrating cells, we have developed a graphical user interface (GUI) capable of automating the tracking of fluorescently labeled nuclei. This GUI provides an intuitive user interface that makes automated tracking accessible to researchers with no image‐processing experience or familiarity with particle‐tracking approaches. Using this GUI, users can interactively determine a minimum of four parameters to identify fluorescently labeled cells and automate acquisition of cell trajectories. Additional features allow for batch processing of numerous time‐lapse images, curation of unwanted tracks, and subsequent statistical analysis of tracked cells. Statistical outputs allow users to evaluate migratory phenotypes, including cell speed, distance, displacement, and persistence, as well as measures of directional movement, such as forward migration index (FMI) and angular displacement. © 2017 by John Wiley & Sons, Inc.
In Vitro Reconstitution of the Endoplasmic Reticulum
Authors: Ferencz Csilla‐Maria , Guigas Gernot , Veres Andreas , Neumann Brigitte , Stemmann Olaf , Weiss Matthias
Published: September 2017

Reconstitution of cellular organelles in vitro offers the possibility to perform quantitative and qualitative experiments in a controlled environment that cannot be done with the same accuracy in living cells. Following a previous report, the subsequent list of protocols describes how to reconstitute and quantify a tubular ER network in vitro based on purified microsomes from culture cells and cytosol from Xenopus laevis egg extracts. Biological material preparation and reconstitution assays require mostly basic laboratory instrumentation and chemicals, and can be executed without any specific training, making them appealing to a wide range of laboratories. Moreover, to promote conditions that are markedly more reflective of in vivo environments, this method describes for the first time in the literature, the purification of microsomes from HeLa cells in some detail. Basic Protocol 1 in this article describes the reconstitution process on different substrates including the associated fluorescence imaging process. Purification of ER microsomes and cytosol, both of which are needed for this approach, are described in detail in Support Protocols 1 and 2, respectively. Coating of surfaces with polyacrylamide gels is described in Support Protocol 3. Basic Protocol 2 outlines how to segment and skeletonize fluorescence images of ER networks, and how to quantify segment lengths between the network's branching points. The described quantitative evaluation provides a meaningful approach to analyze the topology and geometry of organelle structures. © 2017 by John Wiley & Sons, Inc.
Detecting the Interaction of Peptide Ligands with Plant Membrane Receptors
Authors: Hind Sarah Refi , Hoki Jason S. , Baccile Joshua A. , Boyle Patrick C. , Schroeder Frank C. , Martin Gregory B.
Published: September 2017

The field of plant receptor biology has rapidly expanded in recent years, however the demonstration of direct interaction between receptor‐ligand pairs remains a challenge. Click chemistry has revolutionized small molecule research but lacks popularity in plant research. Here we describe a method that tests for the direct physical interaction of a candidate receptor protein and a peptide ligand. This protocol describes the generation of the ligand probe, transient expression of a receptor protein, enrichment of membrane‐bound receptors, photo‐crosslinking and click chemistry‐mediated reporter addition, and detection of the receptor‐ligand complex. Copper‐based click chemistry confers several advantages, including the versatility to use almost any azide‐containing reporter molecule for detection or visualization of the complex and enables addition of the reporter molecule after receptor‐ligand binding which reduces the need for bulky ligand modifications that could interfere with the interaction. © 2017 by John Wiley & Sons, Inc.
Improved Genetic Transformation of Sugarcane (Saccharum spp.) Embryogenic Callus Mediated by Agrobacterium tumefaciens
Authors: Basso Marcos Fernando , da Cunha Bárbara Andrade Dias Brito , Ribeiro Ana Paula , Martins Polyana Kelly , de Souza Wagner Rodrigo , de Oliveira Nelson Geraldo , Nakayama Thiago Jonas , Augusto das Chagas Noqueli Casari Raphael , Santiago Thais Ribeiro , Vinecky Felipe , Cançado Letícia Jungmann , de Sousa Carlos Antônio Ferreira , de Oliveira Patricia Abrão , de Souza Silvana Aparecida Creste Dias , Cançado Geraldo Magela de Almeida , Kobayashi Adilson Kenji , Molinari Hugo Bruno Correa
Published: September 2017

Sugarcane (Saccharum spp.) is a monocotyledonous semi‐perennial C4 grass of the Poaceae family. Its capacity to accumulate high content of sucrose and biomass makes it one of the most important crops for sugar and biofuel production. Conventional methods of sugarcane breeding have shown several limitations due to its complex polyploid and aneuploid genome. However, improvement by biotechnological engineering is currently the most promising alternative to introduce economically important traits. In this work, we present an improved protocol for Agrobacterium tumefaciens‐mediated transformation of commercial sugarcane hybrids using immature top stalk‐derived embryogenic callus cultures. The callus cultures are transformed with preconditioned A. tumefaciens carrying a binary vector that encodes expression cassettes for a gene of interest and the bialaphos resistance gene (bar confers resistance to glufosinate‐ammonium herbicide). This protocol has been used to successfully transform a commercial sugarcane cultivar, SP80‐3280, highlighting: (i) reduced recalcitrance and oxidation; (ii) high yield of embryogenic callus; (iii) improved selection; and (iv) shoot regeneration and rooting of the transformed plants. Altogether, these improvements generated a transformation efficiency of 2.2%. This protocol provides a reliable tool for a routine procedure for sugarcane improvement by genetic engineering. © 2017 by John Wiley & Sons, Inc.
Isolation of Novel MAMP‐like Activities and Identification of Cognate Pattern Recognition Receptors in Arabidopsis thaliana Using Next‐Generation Sequencing (NGS)–Based Mapping
Authors: Fan Li , Chae Eunyoung , Gust Andrea A. , Nürnberger Thorsten
Published: September 2017

Activation of pattern‐triggered plant immunity requires recognition of microbe‐derived molecular patterns (MAMPs) by plant‐encoded pattern recognition receptors (PRRs). Many plant PRRs are found in selected plant genera only. Transfer of single PRRs or of cassettes expressing several PRRs (PRR stacking) across plant genus boundaries offers the potential to boost disease resistance by improving pathogen recognition features in economically important crop plants. The success of such an approach is most dependent on the availability of a large number of plant PRRs. Here, an efficient method for the identification of novel PRRs in the model plant Arabidopsis thaliana (hereafter, Arabidopsis for simplicity) is described. This method takes advantage of natural variation in microbial pattern sensitivity among hundreds of Arabidopsis accessions currently available. Identification of pattern‐sensitive as well as pattern‐insensitive accessions facilitates next‐generation sequencing (NGS)–assisted mapping of PRRs. This approach is potentially applicable to the identification of PRRs that recognize patterns of any chemical nature. © 2017 by John Wiley & Sons, Inc.
Identifying Patients With High Data Completeness to Improve Validity of Comparative Effectiveness Research in Electronic Health Records Data
Authors: Lin Kueiyu Joshua , Singer Daniel E. , Glynn Robert J. , Murphy Shawn N. , Lii Joyce , Schneeweiss Sebastian
Published: September 2017

Electronic health record (EHR)‐discontinuity, i.e., having medical information recorded outside of the study EHR system, is associated with substantial information bias in EHR‐based comparative effectiveness research (CER). We aimed to develop and validate a prediction model identifying patients with high EHR‐continuity to reduce this bias. Based on 183,739 patients aged ≥65 in EHRs from two US provider networks linked with Medicare claims data from 2007–2014, we quantified EHR‐continuity by mean proportion of encounters captured (MPEC) by the EHR system. We built a prediction model for MPEC using one EHR system as training and the other as the validation set. Patients with top 20% predicted EHR‐continuity had 3.5–5.8‐fold smaller misclassification of 40 CER‐relevant variables, compared to the remaining study population. The comorbidity profiles did not differ substantially by predicted EHR‐continuity. These findings suggest that restriction of CER to patients with high predicted EHR‐continuity may confer a favorable validity to generalizability trade‐off.
PhenoGraph and viSNE facilitate the identification of abnormal T‐cell populations in routine clinical flow cytometric data
Authors: DiGiuseppe Joseph A. , Cardinali Jolene L. , Rezuke William N. , Pe'er Dana
Published: September 2017

Background

Flow cytometric identification of neoplastic T‐cell populations is complicated by the wide range of phenotypic abnormalities in T‐cell neoplasia, and the diverse repertoire of reactive T‐cell phenotypes. We evaluated whether a recently described clustering algorithm, PhenoGraph, and dimensionality‐reduction algorithm, viSNE, might facilitate the identification of abnormal T‐cell populations in routine clinical flow cytometric data.

Methods

We applied PhenoGraph and viSNE to peripheral blood mononuclear cells labeled with a single 8‐color T/NK‐cell antibody combination. Individual peripheral blood samples containing either a T‐cell neoplasm or reactive lymphocytosis were analyzed together with a cohort of 10 normal samples, which established the location and identity of normal mononuclear‐cell subsets in viSNE displays.

Results

PhenoGraph‐derived subpopulations from the normal samples formed regions of phenotypic similarity in the viSNE display describing normal mononuclear‐cell subsets, which correlated with those obtained by manual gating (r2 = 0.99, P < 0.0001). In 24 of 24 cases of T‐cell neoplasia with an aberrant phenotype, compared with 4 of 17 cases of reactive lymphocytosis (P = 1.4 × 10−7, Fisher Exact test), PhenoGraph‐derived subpopulations originating exclusively from the abnormal sample formed one or more distinct phenotypic regions in the viSNE display, which represented the neoplastic T cells, and reactive T‐cell subpopulations not present in the normal cohort, respectively. The numbers of neoplastic T cells identified using PhenoGraph/viSNE correlated with those obtained by manual gating (r2 = 0.99; P < 0.0001).

Conclusions

PhenoGraph and viSNE may facilitate the identification of abnormal T‐cell populations in routine clinical flow cytometric data. © 2017 Clinical Cytometry Society
Association between insulin‐induced weight change and CVD mortality: Evidence from a historic cohort study of 18,814 patients in UK primary care
Authors: Anyanwagu Uchenna , Mamza Jil , Donnelly Richard , Idris Iskandar
Published: September 2017

Background

This study explores the association of insulin‐induced weight (wt) gain on cardiovascular outcomes and mortality among patients with type 2 diabetes (T2D) following insulin initiation using real‐world data.

Methods

A historical cohort study was performed in 18,814 adults with insulin‐treated T2D derived from the UK The Health Improvement Network database. Based on the average weight change of 5 kg, 1 year postinsulin initiation, patients were grouped into 5 categories (>5 kg wt loss; 1.0‐5.0 kg wt loss; no wt change; 1.0‐5.0 kg wt gain; >5.0 kg wt gain) and followed‐up for 5 years. Cox proportional hazard models and Kaplan‐Meier estimators were fitted to estimate the hazards of a 3‐point composite of nonfatal myocardial infarction, stroke, and all‐cause mortality between categories.

Results

The median age was 62.8 (IQR: 52.3–71.8) years, HbA1c: 8.6% (IQR: 7.4–9.8) and mean BMI: 31.8 (6.5) kg/m2. The 5 year probability of survival differed significantly within the wt‐change categories (log‐rank test P value = .0005). Only 1963 composite events occurred. Compared with the weight‐neutral group, the risk of composite events was 31% greater in the >5 kg wt‐loss group (aHR: 1.31; 95% CI: 1.02, 1.68), the same in the 1.0 to 5.0 kg wt‐gain category, but nonsignificantly increased in the 1.0 to 5.0 kg wt loss (15%) and >5.0 kg wt gain (13%) categories, respectively. In the obese subgroup, this risk was 50% (aHR: 1.50, 95% CI: 1.08‐2.08) more in the >5 kg weight‐loss group compared with the weight‐neutral group.

Conclusion

Insulin‐induced weight gain did not translate to adverse cardiovascular outcomes and mortality in patients with T2D. These data provide reassurance on the cardiovascular safety of insulin patients with T2D.
A budget of algal production and consumption by herbivorous fish in an herbivore fisheries management area, Maui, Hawaii
Authors: Kelly Emily L. A. , Eynaud Yoan , Williams Ivor D. , Sparks Russell T. , Dailer Meghan L. , Sandin Stuart A. , Smith Jennifer E.
Published: September 2017

The relative rates of carbon fixed by primary producers vs. consumption by primary consumers shape the community of organisms in ecosystems. On coral reefs, it is important to understand the demography of algae, given known competitive dynamics with reef‐building corals. Numerous studies have shown that fleshy algal abundance is enhanced in the absence of top‐down control by herbivores on coral reefs. However, fewer studies have quantified and compared rates of production by the benthic fleshy algal community and consumption by herbivores. Here, we estimate a budget for fleshy algal growth and herbivorous fish consumption on a Hawaiian coral reef based upon integration of field‐measured and taxonomically specific variables. Data were collected at Kahekili Herbivore Fisheries Management Area, Maui, established in 2009, in which taking of herbivorous fish and urchins is prohibited. Daily algal production was determined by quantifying benthic community composition, standing stock of algal biomass, and growth rates of common algal components. Consumption was determined using distributions of biomass and size classes of herbivorous fish species, consumption rates of herbivores on different algal species, and herbivore bite sizes. Our results show that throughout the first five years of herbivore protection, algal production consistently exceeded the grazing capacity of the herbivorous fish assemblage, but by a diminishing margin since 2010 (consumption 20.8% of production) to the end of the study in 2014 (consumption 67.0% of production). Further, larger size classes of herbivorous fishes in the scraper/excavator herbivore guild contributed more to consumption in later years, which could have additional feedbacks that promote reef‐building taxa. Projecting the budget using data from a neighboring decades‐long protected herbivorous fish assemblage indicated that the production and consumption budget for Kahekili could become balanced in future with continued management.
Is initial post‐disturbance regeneration indicative of longer‐term trajectories?
Authors: Gill Nathan S. , Jarvis Daniel , Veblen Thomas T. , Pickett Steward T. A. , Kulakowski Dominik
Published: September 2017

The ability to estimate and model future vegetation dynamics is a central focus of contemporary ecology and is essential for understanding future ecological trajectories. It is therefore critical to understand when the influence of initial post‐disturbance regeneration versus stochastic processes dominates long‐term post‐disturbance ecological processes. Often, conclusions about post‐disturbance dynamics are based upon initial regeneration in the years immediately after disturbances. However, the degree to which initial post‐disturbance regeneration indicates longer‐term trends is likely to be contingent on the types, intensities, and combinations of disturbances, as well as pre‐disturbance ecosystem structure and composition. Our relatively limited understanding of why initial post‐disturbance regeneration is sometimes a poor predictor of future ecosystem trajectories represents a critical gap in post‐disturbance ecological forecasting. We studied the composition and density of regeneration of tree species following wind blowdown in 1997, wildfire in 2002, and compounded disturbances by blowdown and wildfire in subalpine forests of Colorado. We examined regeneration of Picea engelmannii, Abies lasiocarpa, Pinus contorta, and Populus tremuloides in 180 permanent plots across 12 sites (classified by pre‐disturbance age and composition) in 2003, 2010, and 2015. At sites that were blown down but not burned, regeneration was dense and dominated by Picea and Abies. At these sites, regeneration observed from 2003 to 2005 (hereafter initial regeneration) was also highly predictive of regeneration 5–10 yr later. In contrast, at sites that were burned and sites that were blown down and burned, regeneration was less dense and dominated by a mix of species. At these sites, initial regeneration was a poor predictor of longer‐term trends as species dominance and overall density fluctuated over the 13‐yr period. These findings call into question our ability to confidently predict ecosystem trajectories based upon observations made in the years immediately after large, severe disturbances such as wildfires and compounded disturbances. As compounded disturbances become more common under climatically driven changes in disturbance regimes, post‐disturbance ecosystem trajectories may become increasingly stochastic and unpredictable.
Bayesian networks elucidate interactions between fire and other drivers of terrestrial fauna distributions
Authors: Hradsky Bronwyn A. , Penman Trent D. , Ababei Dan , Hanea Anca , Ritchie Euan G. , York Alan , Di Stefano Julian
Published: September 2017

Fire is a major driver of community composition and habitat structure and is extensively used as an ecological management tool in flammable landscapes. Interactions between fire and other processes that affect animal distributions, however, cause variation in faunal responses to fire and limit our ability to identify appropriate fire management regimes for biodiversity conservation. Bayesian networks (BNs) have not previously been used to examine terrestrial faunal distributions in relation to fire, but offer an alternative statistical framework for modeling complex environmental relationships as they explicitly capture interactions between predictor variables. We developed a conceptual model of the interactions between drivers of faunal distributions in fire‐affected landscapes, and then used a non‐parametric BN modeling approach to describe and quantify these relationships for a suite of terrestrial native mammal species. We also tested whether BNs could be used to predict these species’ distributions using only remote‐sensed or mapped variables. Data were collected at 113 sites across 47,000 ha of continuous eucalypt forest in the Otway Ranges, southeastern Australia; time‐since‐fire (TSF) ranged from six months to 74 yr. Habitat complexity increased with TSF and forest wetness. Critical‐weight‐range (35–5500 g) marsupials and rodents were generally more likely to occur at long unburnt sites with high habitat complexity, and in wetter forest types. In contrast, large grazers and browsers preferred less complex habitats and younger or drier forest. Species occurrences were more strongly affected by habitat complexity than TSF, coarse woody debris cover, or invasive predator (Vulpes vulpesorFelis catus) occurrence. Bayesian network models effectively discriminated between the presence and absence of most native mammal species, even when only provided with data on remote‐sensed or mapped variables (i.e., without field‐assessed data such as habitat complexity). Non‐parametric BNs are an effective technique for explicitly modeling the complex and context‐dependent influence of fire history on faunal distributions, and may reduce the need to collect extensive field data on habitat structure and other proximate drivers.
Evaluation of an acute oral gavage method for assessment of pesticide toxicity in terrestrial amphibians
Authors: Fort Douglas J. , Mathis Michael B. , Kee Faith , Whatling Paul , Clerkin David , Staveley Jane , Habig Clifford
Published: September 2017

Development of an acute oral toxicity test with a terrestrial-phase amphibian was considered necessary to remove the uncertainty within the field of agrochemical risk assessments. The bullfrog (Lithobates catesbeianus) was selected for use as it is a representative of the family Ranidae and historically this species has been used as an amphibian test model species. Prior to definitive study, oral gavage methods were developed with fenthion and tetraethyl pyrophosphate. Dimethoate and malathion were subsequently tested with both male and female juvenile bullfrogs in comprehensive acute oral median lethal dose (LD50) studies. Juvenile bullfrogs were administered a single dose of the test article via oral gavage of a single gelatin capsule of dimethoate technical (dimethoate) or neat liquid Fyfanon® Technical (synonym malathion), returned to their respective aquaria, and monitored for survival for 14 d. The primary endpoint was mortality, whereas behavioral responses, food consumption, body weight, and snout–vent length (SVL) were used to evaluate indications of sublethal toxicity (secondary endpoints). Acute oral LD50 values (95% fiducial interval) for dimethoate were 1459 (1176–1810, males) and 1528 (1275–1831, females), and for malathion they were 1829 (1480–2259, males) and 1672 (1280–2183, females) mg active substance/kg body weight, respectively. Based on the results of these studies, the methodology for the acute oral gavage administration of test items to terrestrial-phase amphibians was demonstrated as being a practical method of providing data for risk assessments. Environ Toxicol Chem 2018;37:436–450. © 2017 SETAC
Uptake of radiolabeled 3,3′,4,4′-tetrachlorobiphenyl into Japanese quail egg compartments and embryo following air cell and albumen injection
Authors: Dean Karen M. , Baltos Leah D. , Marcell Allegra M. , Bohannon Meredith E.B. , Iwaniuk Andrew N. , Ottinger Mary Ann
Published: September 2017

The avian embryo is an excellent model for testing adverse developmental effects of environmental chemicals as well as uptake and movement of xenobiotics within the egg compartments. Before incubation at embryonic day 0, 14C 3,3′,4,4′-tetrachlorobiphenyl (14C PCB 77) was injected into Japanese quail eggs either onto the air cell or into the albumen. All egg components were collected on embryonic day 1, 5, or 10, and concentrations of 14C PCB 77 were measured in various egg components (shell, membrane, yolk, albumen, and embryo). The results showed measurable 14C PCB 77 in all egg components, with changing concentrations in each egg component over the course of embryonic development. Specifically, concentrations in the shell content decreased between embryonic days 1 and 10, increased in albumen from embryonic days 1 to 5 and then decreased at embryonic day 10, and increased in both yolk and embryo from embryonic days 1 to 10. Vehicle and injection site both influenced 14C PCB 77 allantoic fluid concentrations, with little effect on other egg components except for the inner shell membrane. The fatty acid vehicle injected into the albumen yielded the highest 14C PCB 77 recovery. These findings demonstrate dynamic movement of toxicants throughout the egg components during avian embryonic development and a steady increase of relatively low levels of 14C PCB 77 in the embryo compared with the yolk, albumen, and shell, suggesting that embryonic uptake (i.e., exposure) mirrors utilization of egg components for nutrition and growth during development. Environ Toxicol Chem 2018;37:126–135. © 2017 SETAC
Gene‐disease associations identify a connectome with shared molecular pathways in human cholangiopathies
Authors: Luo Zhenhua , Jegga Anil G. , Bezerra Jorge A.
Published: September 2017

Cholangiopathies are a diverse group of progressive diseases whose primary cell targets are cholangiocytes. To identify shared pathogenesis and molecular connectivity among the three main human cholangiopathies (biliary atresia [BA], primary biliary cholangitis [PBC], and primary sclerosing cholangitis [PSC]), we built a comprehensive platform of published data on gene variants, gene expression, and functional studies and applied network‐based analytics in the search for shared molecular circuits. Mining the data platform with largest connected component and interactome analyses, we validated previously reported associations and identified essential and hub genes. In addition to disease‐specific modules, we found a substantial overlap of disease neighborhoods and uncovered a group of 34 core genes that are enriched for immune processes and abnormal intestine/hepatobiliary mouse phenotypes. Within this core, we identified a gene subcore containing signal transduction and activator of transcription 3, interleukin‐6, tumor necrosis factor, and forkhead box P3 prominently placed in a regulatory connectome of genes related to cellular immunity and fibrosis. We also found substantial gene enrichment in the advanced glycation endproduct/receptor for advanced glycation endproducts (RAGE) pathway and showed that RAGE activation induced cholangiocyte proliferation. Conclusion: Human cholangiopathies share pathways enriched by immunity genes and a molecular connectome that links different pathogenic features of BA, PBC, and PSC. (Hepatology 2018;67:676‐689).
Efficacy and safety of glecaprevir/pibrentasvir in Japanese patients with chronic genotype 2 hepatitis C virus infection
Authors: Toyoda Hidenori , Chayama Kazuaki , Suzuki Fumitaka , Sato Ken , Atarashi Tomofumi , Watanabe Tsunamasa , Atsukawa Masanori , Naganuma Atsushi , Notsumata Kazuo , Osaki Yukio , Nakamuta Makoto , Takaguchi Koichi , Saito Satoru , Kato Koji , Pugatch David , Burroughs Margaret , Redman Rebecca , Alves Katia , Pilot‐Matias Tami J. , Oberoi Rajneet K. , Fu Bo , Kumada Hiromitsu
Published: September 2017

Glecaprevir (nonstructural protein 3/4A protease inhibitor) and pibrentasvir (nonstructural protein 5A inhibitor) (G/P), a coformulated once‐daily, all oral, ribavirin (RBV)‐free, direct‐acting antiviral regimen, was evaluated for safety and efficacy in hepatitis C virus genotype 2 (GT2)–infected Japanese patients, including those with compensated cirrhosis. CERTAIN‐2 is a phase 3, open‐label, multicenter study assessing the safety and efficacy of G/P (300/120 mg) once daily in treatment‐naive and interferon ± RBV treatment–experienced Japanese patients without cirrhosis but with GT2 infection. Patients were randomized 2:1 to receive 8 weeks of G/P (arm A) or 12 weeks of sofosbuvir (400 mg once daily) + RBV (600‐1000 mg weight‐based, twice daily) (arm B). The primary endpoint was noninferiority of G/P compared to sofosbuvir + RBV by assessing sustained virologic response at posttreatment week 12 (SVR12) among patients in the intent‐to‐treat population. SVR12 was also assessed in treatment‐naive and interferon ± RBV treatment‐experienced patients with GT2 infection and compensated cirrhosis who received G/P for 12 weeks in the CERTAIN‐1 study. A total of 136 patients were enrolled in CERTAIN‐2. SVR12 was achieved by 88/90 (97.8%) patients in arm A and 43/46 (93.5%) patients in arm B. No patient in arm A experienced virologic failure, while 2 did in arm B. The primary endpoint was achieved. In CERTAIN‐1, 100% (18/18) of GT2‐infected patients with compensated cirrhosis achieved SVR12. Treatment‐emergent serious adverse events were experienced by 2 patients without cirrhosis in each arm and no patient with cirrhosis. Conclusion: The results demonstrate high efficacy and favorable tolerability of G/P in GT2‐infected Japanese patients. (Hepatology 2018;67:505‐513).
Evaluation of Several Molybdenum and Ruthenium Catalysts for the Metathesis Homocoupling of 3‐Methyl‐1‐Butene
Authors: Sues Peter E. , Bukhryakov Konstantin V. , Schrock Richard R.
Published: September 2017

We synthesized Mo(NC6F5)(CHCMe2Ph)(TPPO)(PPhMe2)Cl (TPPO = 2,3,5,6‐tetraphenylphenoxide), Mo(NC6F5)(CHCMe2Ph)(TTBTO)(PPhMe2)Cl (TTBTO = 2,6‐di(3′,5′‐di‐tert‐butylphenyl)phenoxide), and Mo(NC6F5)(CHCMe2Ph)(TPPO)(PPhMe2)(CF3Pyr) (CF3Pyr = 3,4‐bistrifluoromethylpyrrolide), in order to evaluate them as catalysts for the homocoupling of 3‐methyl‐1‐butene. They were compared with Mo(NC6F5)(CHCMe2Ph)(HMTO)(PPhMe2)Cl (HMTO = 2,6‐dimesitylphenoxide), Mo(NC6F5)(CHCMe2Ph)(HIPTO)(PPhMe2)Cl (HIPTO = 2,6‐di(2′,4′,6′‐triisopropylphenyl)phenoxide), and several other Mo and Ru catalysts. In the best cases turnover numbers (TONs) of 400 – 700 were observed for the homocoupling of 3‐methyl‐1‐butene in a closed vessel (ethylene not removed).
Maternal health outcomes following the implementation of Janani Shishu Suraksha Yojana (JSSY) at a tertiary healthcare center in Rajasthan
Authors: Mahala Urmila , Mehta Seema
Published: September 2017

Objectives

To assess the impact of the Janani Shishu Suraksha Yojana (JSSY) initiative on maternal health in terms of the number of prenatal checkup registrations, institutional deliveries, and maternal deaths. The aim of JSSY was to reduce maternal and infant mortality by promoting institutional delivery.

Methodology

The present retrospective descriptive study included data from all institutional deliveries that occurred at the Department of Obstetrics and Gynecology, Mahila Chikitsalaya, SMS Medical College Jaipur, India, both before (between January 1, 2008, and December 31, 2011) and after the implementation of JSSY (between January 1, 2012, and December 31, 2015). Numbers of prenatal outpatient department registrations, institutional deliveries, and maternal deaths per year were compared between the periods using the Mann‐Whitney U test.

Results

Increases in the mean number of prenatal outpatient department registrations (P=0.021) and institutional deliveries (P=0.021) were recorded following the implementation of JSSY; a non‐significant decrease in the maternal mortality ratio was also observed (P=0.248).

Conclusion

JSSY appeared to be an effective program in improving maternal health; use of prenatal care and institutional deliveries increased following its implementation.
Learning curve for the single‐incision suburethral sling procedure for female stress urinary incontinence
Authors: Spelzini Federico , Frigerio Matteo , Regini Cristina , Palmieri Stefania , Manodoro Stefano , Milani Rodolfo
Published: September 2017

Objective

To evaluate the learning curve for the single‐incision sling (SIS) procedure in a single‐surgeon case series.

Methods

Data were retrospectively analyzed from women with non‐recurrent symptomatic stress urinary incontinence confirmed by urodynamics and treated at San Gerardo Hospital, Monza, Italy, between October 2008 and November 2015. All women underwent the SIS procedure using a MiniArc sling fitted by the same urogynecologic surgeon. Blood loss, operative time, and complications were recorded. At follow‐up visits, objective cure was assessed with a stress test and subjective outcome was determined by International Consultation on Incontinence Questionnaire‐Short Form and Patient Global Impression of Improvement scores. Postoperative overactive bladder syndrome and voiding symptoms were recorded.

Results

In total, 192 patients underwent the SIS procedure. Estimated blood loss, operative time, and complication rate were not influenced by the number of procedures performed. Among 191 patients who completed 12 months of follow‐up, neither objective nor subjective outcomes at longest follow‐up available were influenced by surgeon's experience. Conversely, de novo overactive bladder syndrome (P<0.001) and voiding symptoms (P=0.029) decreased with increasing experience.

Conclusion

SIS showed encouraging objective and subjective outcomes with a minimal complication rate even at the beginning of the learning curve. However, increasing experience reduced postoperative voiding symptoms and overactive bladder syndrome.
Sustained release of multicomponent platelet‐rich plasma proteins from hydrolytically degradable PEG hydrogels
Authors: Jain Era , Sheth Saahil , Dunn Andrew , Zustiak Silviya P. , Sell Scott A.
Published: September 2017

Platelet‐rich plasma (PRP), an autologous blood derived product is a concentrated mix of multiple growth factors and cytokines. Direct injections of PRP are clinically used for treatment of various musculoskeletal disorders and in wound healing. However, PRP therapy has met with limited clinical success possibly due to unpredictable and premature bolus delivery of PRP growth factors. The objective of this study was to predictably control the bioavailability of PRP growth factors using a hydrolytically degradable polyethylene glycol (PEG) hydrogel. We used a step‐growth polymerization based on a Michael‐type addition reaction between a 6‐arm PEG‐acrylate and a dithiol crosslinker, which led to the formation of a homogenous hydrogel network under mild, physiologically relevant conditions. Specifically, to model the release of multicomponent PRP through PEG hydrogels, we examined bulk diffusion of PRP as well as model proteins in a size range corresponding to that of growth factors found in PRP. Our results indicated that protein size and hydrogel degradation controlled diffusion of all proteins and that secondary structure of proteins encapsulated during gelation remained unaffected post‐release. Analysis of specific PRP proteins released from the hydrogel showed sustained release until complete hydrogel degradation. PRP released from hydrogels promoted proliferation of human dermal fibroblast, indicating retained bioactivity upon encapsulation and release. The versatile hydrogel system holds clinical potential as a therapeutic drug delivery depot of multicomponent mixtures like PRP. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3304–3314, 2017.
Control of cell growth on 3D‐printed cell culture platforms for tissue engineering
Authors: Tan Zhikai , Liu Tong , Zhong Juchang , Yang Yikun , Tan Weihong
Published: September 2017

Biocompatible tissue growth has excellent prospects for tissue engineering. These tissues are built over scaffolds, which can influence aspects such as cell adhesion, proliferation rate, morphology, and differentiation. However, the ideal 3D biological structure has not been developed yet. Here, we applied the electro‐hydrodynamic jet (E‐jet) 3D printing technology using poly‐(lactic‐co‐glycolic acid, PLGA) solution to print varied culture platforms for engineered tissue structures. The effects of different parameters (electrical voltage, plotting speed, and needle sizes) on the outcome were investigated. We compared the biological compatibility of the 3D printed culture platforms with that of random fibers. Finally, we used the 3D‐printed PLGA platforms to culture fibroblasts, the main cellular components of loose connective tissue. The results show that the E‐jet printed platforms could guide and improve cell growth. These highly aligned fibers were able to support cellular alignment and proliferation. Cell angle was consistent with the direction of the fibers, and cells cultured on these fibers showed a much faster migration, potentially enhancing wound healing performance. Thus, the potential of this technology for 3D biological printing is large. This process can be used to grow biological scaffolds for the engineering of tissues. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3281–3292, 2017.
Surface topography of silicon nitride affects antimicrobial and osseointegrative properties of tibial implants in a murine model
Authors: Ishikawa Masahiro , de Mesy Bentley Karen L. , McEntire Bryan J. , Bal B. Sonny , Schwarz Edward M. , Xie Chao
Published: September 2017

While silicon nitride (Si3N4) is an antimicrobial and osseointegrative orthopaedic biomaterial, the contribution of surface topography to these properties is unknown. Using a methicillin‐resistant strain of Staphylococcus aureus (MRSA), this study evaluated Si3N4 implants in vitro utilizing scanning electron microscopy (SEM) with colony forming unit (CFU) assays, and later in an established in vivo murine tibia model of implant‐associated osteomyelitis. In vitro, the “as‐fired” Si3N4 implants displayed significant reductions in adherent bacteria versus machined Si3N4 (2.6 × 104 vs. 8.7 × 104 CFU, respectively; p < 0.0002). Moreover, SEM imaging demonstrated that MRSA cannot directly adhere to native as‐fired Si3N4. Subsequently, a cross‐sectional study was completed in which sterile or MRSA contaminated as‐fired and machined Si3N4 implants were inserted into the tibiae of 8‐week old female Balb/c mice, and harvested on day 1, 3, 5, 7, 10, or 14 post‐operatively for SEM. The findings demonstrated that the antimicrobial activity of the as‐fired implants resulted from macrophage clearance of the bacteria during biofilm formation on day 1, followed by osseointegration through the apparent recruitment of mesenchymal stem cells on days 3–5, which differentiated into osteoblasts on days 7–14. In contrast, the antimicrobial behavior of the machined Si3N4 was due to repulsion of the bacteria, a phenomenon that also limited osteogenesis, as host cells were also unable to adhere to the machined surface. Taken together, these results suggest that the in vivo biological behavior of Si3N4 orthopaedic implants is driven by critical features of their surface nanotopography. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3413–3421, 2017.
RETRACTED: Kaempferol increases apoptosis in human acute promyelocytic leukemia cells and inhibits multidrug resistance genes
Authors: Moradzadeh Maliheh , Tabarraei Alijan , Sadeghnia Hamid Reza , Ghorbani Ahmad , Mohamadkhani Ashraf , Erfanian Saiedeh , Sahebkar Amirhossein
Published: September 2017

Acute promyelocytic leukemia (APL) is one of the most life‐threatening hematological malignancies. Defects in the cell growth and apoptotic pathways are responsible for both disease pathogenesis and treatment resistance. Therefore, pro‐apoptotic agents are potential candidates for APL treatment. Kaempferol is a flavonoid with antioxidant and anti‐tumor properties. This study was designed to investigate the cytotoxic, pro‐apoptotic, and differentiation‐inducing effects of kaempferol on HL‐60 and NB4 leukemia cells. Resazurin assay was used to determine cell viability following treatment with kaempferol (12.5‐100 μM) and all‐trans retinoic acid (ATRA; 10 μM; used as a positive control). Apoptosis and differentiation were also detected using propidium iodide and NBT staining techniques, respectively. Furthermore, the expression levels of genes involved in apoptosis (PI3 K, AKT, BCL2, BAX, p53, p21, PTEN, CASP3, CASP8, and CASP9), differentiation (PML‐RAR and HDAC1), and multi‐drug resistance (ABCB1 and ABCC1) were determined using quantitative real‐time PCR. The protein expressions of Bax/Bcl2 and casp3 were confirmed using Western blot. The results showed that kaempferol decreased cell viability and increased subG1 population in the tested leukemic cells. This effect was associated with decreased expression of Akt, BCL2, ABCB1, and ABCC1 genes, while the expression of CASP3 and BAX/BCL‐2 ratio were significantly increased at both gene and protein levels. Kaempferol promoted apoptosis and inhibited multidrug resistance in a concentration‐dependent manner, without any differential effect on leukemic cells. In conclusion, this study suggested that kaempferol may be utilized as an appropriate alternative for ATRA in APL patients.