bluuebunny/crossref_metadata_2025 huggingface dataset:
3D scanning and printing skeletal tissues for anatomy education
Authors: Thomas Daniel B. , Hiscox Jessica D. , Dixon Blair J. , Potgieter Johan
Published: May 2016

Detailed anatomical models can be produced with consumer‐level 3D scanning and printing systems. 3D replication techniques are significant advances for anatomical education as they allow practitioners to more easily introduce diverse or numerous specimens into classrooms. Here we present a methodology for producing anatomical models in‐house, with the chondrocranium cartilage from a spiny dogfish (Squalus acanthias) and the skeleton of a cane toad (Rhinella marina) as case studies. 3D digital replicas were produced using two consumer‐level scanners and specimens were 3D‐printed with selective laser sintering. The fidelity of the two case study models was determined with respect to key anatomical features. Larger‐scale features of the dogfish chondrocranium and frog skeleton were all well‐resolved and distinct in the 3D digital models, and many finer‐scale features were also well‐resolved, but some more subtle features were absent from the digital models (e.g. endolymphatic foramina in chondrocranium). All characters identified in the digital chondrocranium could be identified in the subsequent 3D print; however, three characters in the 3D‐printed frog skeleton could not be clearly delimited (palatines, parasphenoid and pubis). Characters that were absent in the digital models or 3D prints had low‐relief in the original scanned specimen and represent a minor loss of fidelity. Our method description and case studies show that minimal equipment and training is needed to produce durable skeletal specimens. These technologies support the tailored production of models for specific classes or research aims.
Therapeutic effects of Wharton jelly‐derived mesenchymal stem cells on rat abortion models
Authors: Chen Xiaojing , Yang Xiaoqing , Wu Rongrong , Chen Weiwei , Xie Huihui , Qian Xia , Zhang Yuquan
Published: May 2016

Aim

The aim of this study was to investigate the therapeutic effects and mechanism of Wharton jelly‐derived mesenchymal stem cells (WJ‐MSC) in a spontaneous‐abortion rat model.

Methods

An abortion model was established using intravenous injection of bromocriptine. WJ‐MSC were isolated and cultured from Wharton jelly of the human umbilical cord. The pathologic changes of placenta decidual tissues were observed after hematoxylin–eosin staining. The embryo absorption rate was counted. The protein and mRNA levels of interleukin (IL)‐10, interferon (IFN)‐γ and IL‐17 in each group were tested by enzyme‐linked immunosorbent assay, Western blot and quantitative real‐time polymerase chain reaction.

Results

After bromocriptine injection, decidual cells degenerated and the connections between them loosened. Furthermore, some of the decidual cells were necrotic and the nuclei had disappeared. However, the transplantation of a large population of WJ‐MSC prevented these damages from occurring. The changes of levels of IL‐10, IFN‐γ and IL‐17 in serum and placenta decidual tissues induced by bromocriptine were well restored by a high, but not a low, dose of WJ‐MSC engraft.

Conclusion

WJ‐MSC can ameliorate early pregnancy loss. The mechanism may be related to its upregulation of IL‐10 and downregulation of IFN‐γ and IL‐17.
Examining Discrimination, Ethnic‐Racial Identity Status, and Youth Public Regard Among Black, Latino, and White Adolescents
Authors: Douglass Sara , Umaña‐Taylor Adriana J.
Published: May 2016

How positively adolescents believe others feel about their ethnic‐racial group (i.e., public regard) is an important part of their ethnic‐racial identity (ERI), which is likely informed by contextual and individual factors. Using cluster analyses to generate ERI statuses among Black, Latino, and White adolescents (n = 1,378), we found that associations between peer versus adult discrimination and public regard varied across ERI status and ethnic‐racial group. However, among all adolescents, an achieved ERI (i.e., having explored ethnicity‐race and having a clear sense about its personal meaning) buffered the negative association between adult discrimination and public regard, but not between peer discrimination and public regard. Implications for understanding the interplay between contextual and individual factors for public regard are discussed.
Neonatal encephalopathy in New Zealand: Demographics and clinical outcome
Authors: Battin M , Sadler L , Masson V , Farquhar C ,   on behalf of the Neonatal Encephalopathy Working Group of the PMMRC
Published: May 2016

Aim

To establish the incidence of moderate to severe neonatal encephalopathy (NE) in term infants from New Zealand and to document demographic characteristics and neonatal outcomes.

Methods

Cases were reported monthly via the New Zealand Paediatric Surveillance Unit (NZPSU). Data were collected from paediatricians for neonatal items and lead maternity carers for pregnancy and birth details. Term neonatal deaths in the Perinatal and Maternal Mortality Review Committee dataset that were because of hypoxia and/or neonatal deaths from hypoxic ischaemic encephalopathy were added to the cases identified via the NZPSU, if they had not previously been ascertained.

Results

For the period January 2010 to December 2012, there were 227 cases, equivalent to a rate of 1.30/1000 term births (95% CI 1.14–1.48). Rates of NE were high in babies of Pacific and Indian mothers but only reached statistical significance for the comparison between Pacific and NZ European. There was also a significant increase in NE rates with increasing deprivation.

Resuscitation at birth was initiated for 209 (92.1%) infants with NE. Mechanical ventilation was required, following neonatal unit admission, in 171 (75.3%) infants. Anticonvulsants were used in 157 (69.2%) infants with phenobarbitone (65.6%), phenytoin (14.5%) and benzodiazapines (21.1%), the most common. Cooling was induced in 168 infants (74%) with 145 (86.3%) reported as commenced within a 6‐h window.

Conclusions

The rate of NE in New Zealand is consistent with reported international rates. Establishing antecedent factors for NE is an important part of improving care, which may inform strategic efforts to decrease rates of NE.
Outcome of outborn infants at the borderline of viability in Western Australia: A retrospective cohort study
Authors: Thompson Kirsten , Gardiner Jacqueline , Resnick Steven
Published: May 2016

Aim

Preterm infants have a high risk of morbidity and mortality, which increases with decreasing gestational age. Inborn infants (infants born in tertiary perinatal centres) have higher survival and lower morbidity than outborn infants. We aimed to compare short‐term and 1‐year developmental outcomes of outborn infants at the borderline of viability (≥23 to ≤25 + 6 weeks gestation) with a similar cohort of inborn infants in the sole tertiary perinatal centre in Western Australia from 2001 to 2011.

Methods

This was a retrospective cohort study. Outborn infants ≥23 to ≤25 + 6 weeks gestation who survived to be transported to the Neonatal Intensive Care Unit (NICU) in the perinatal centre were contemporaneously matched to the next inborn infant of comparable gestation and birth weight. We compared mortality, morbidity (including intraventricular haemorrhage, necrotising enterocolitis and chronic lung disease) and Griffiths General Quotient scores at 1‐year corrected age.

Results

There were 54 outborn and 519 inborn births in the gestational age range during the study period. Thirty‐five (65%) outborn infants were transported to the NICU. Of the outborn infants, 21/54 (39%) survived to discharge compared with 375/519 (72%) inborn infants. For the 35 outborn infants transported to NICU, 14 (40%) died, compared with 6/35 (17%) of inborn infants. There were no differences in short‐term and developmental outcomes in surviving infants.

Conclusions

Outborn extremely preterm infants <26 weeks gestation have higher mortality than inborn counterparts. However, those transported to a tertiary NICU have similar morbidity and developmental outcomes.
The C‐terminus of amelogenin enhances osteogenic differentiation of human cementoblast lineage cells
Authors: Kunimatsu R. , Yoshimi Y. , Hirose N. , Awada T. , Miyauchi M. , Takata T. , Li W. , Zhu L. , Denbesten P.K. , Tanimoto K.
Published: May 2016

Background and Objectives

Amelogenin proteins are the major constituent of developing extracellular enamel matrix and are believed to have an exclusively epithelial origin. Recent studies have suggested that amelogenins might induce the differentiation and maturation of various cells, including cementoblast lineage cells. However, the residues comprising the active site of amelogenin remain unclear. The purpose of this study was to identify the active site region of amelogenin by studying the effects of amelogenin fragments on the osteogenic differentiation of cementoblasts.

Material and Methods

Amelogenin fragments lacking the C‐terminus (rh163) and N‐terminus (rh128) and a fragment consisting of the C‐terminal region of rh174 (C11 peptide) were synthesized and purified. Human cementoblast lineage cells were cultured in osteogenic differentiation medium and treated with 0, 10, 100 or 1000 ng/mL of rh163, rh128 or C11 peptide. The mRNA levels of bone markers were examined by real‐time polymerase chain reaction analysis. Alkaline phosphatase activity and calcium deposition were also determined. Mineralization was evaluated by alizarin red staining.

Results

The osteogenic differentiation of human cementoblast lineage cells was significantly enhanced by treatment with rh128 or C11 peptide, whereas rh163 had no significant effect as compared with untreated controls.

Conclusions

The C‐terminus of amelogenin promotes the osteogenic differentiation of human cementoblast lineage cells, indicating the possible utility of C11 peptide in periodontal tissue regeneration.
Trichobaris weevils distinguish amongst toxic host plants by sensing volatiles that do not affect larval performance
Authors: Lee Gisuk , Joo Youngsung , Diezel Celia , Lee Eun Ju , Baldwin Ian T. , Kim Sang‐Gyu
Published: May 2016

Herbivorous insects use plant metabolites to inform their host plant selection for oviposition. These host‐selection behaviours are often consistent with the preference–performance hypothesis; females oviposit on hosts that maximize the performance of their offspring. However, the metabolites used for these oviposition choices and those responsible for differences in offspring performance remain unknown for ecologically relevant interactions. Here, we examined the host‐selection behaviours of two sympatric weevils, the Datura (Trichobaris compacta) and tobacco (T. mucorea) weevils in field and glasshouse experiments with transgenic host plants specifically altered in different components of their secondary metabolism. Adult females of both species strongly preferred to feed on D. wrightii rather than on N. attenuata leaves, but T. mucorea preferred to oviposit on N. attenuata, while T. compacta oviposited only on D. wrightii. These oviposition behaviours increased offspring performance: T. compacta larvae only survived in D. wrightii stems and T. mucorea larvae survived better in N. attenuata than in D. wrightii stems. Choice assays with nicotine‐free, JA‐impaired, and sesquiterpene‐over‐produced isogenic N. attenuata plants revealed that although half of the T. compacta larvae survived in nicotine‐free N. attenuata lines, nicotine did not influence the oviposition behaviours of both the nicotine‐adapted and nicotine‐sensitive species. JA‐induced sesquiterpene volatiles are key compounds influencing T. mucorea females’ oviposition choices, but these sesquiterpenes had no effect on larval performance. We conclude that adult females are able to choose the best host plant for their offspring and use chemicals different from those that influence larval performance to inform their oviposition decisions.
A synthesis of genetic connectivity in deep‐sea fauna and implications for marine reserve design
Authors: Baco Amy R. , Etter Ron J. , Ribeiro Pedro A. , von der Heyden Sophie , Beerli Peter , Kinlan Brian P.
Published: May 2016

With anthropogenic impacts rapidly advancing into deeper waters, there is growing interest in establishing deep‐sea marine protected areas (MPAs) or reserves. Reserve design depends on estimates of connectivity and scales of dispersal for the taxa of interest. Deep‐sea taxa are hypothesized to disperse greater distances than shallow‐water taxa, which implies that reserves would need to be larger in size and networks could be more widely spaced; however, this paradigm has not been tested. We compiled population genetic studies of deep‐sea fauna and estimated dispersal distances for 51 studies using a method based on isolation‐by‐distance slopes. Estimates of dispersal distance ranged from 0.24 km to 2028 km with a geometric mean of 33.2 km and differed in relation to taxonomic and life‐history factors as well as several study parameters. Dispersal distances were generally greater for fishes than invertebrates with the Mollusca being the least dispersive sampled phylum. Species that are pelagic as adults were more dispersive than those with sessile or sedentary lifestyles. Benthic species from soft‐substrate habitats were generally less dispersive than species from hard substrate, demersal or pelagic habitats. As expected, species with pelagic and/or feeding (planktotrophic) larvae were more dispersive than other larval types. Many of these comparisons were confounded by taxonomic or other life‐history differences (e.g. fishes being more dispersive than invertebrates) making any simple interpretation difficult. Our results provide the first rough estimate of the range of dispersal distances in the deep sea and allow comparisons to shallow‐water assemblages. Overall, dispersal distances were greater for deeper taxa, although the differences were not large (0.3–0.6 orders of magnitude between means), and imbalanced sampling of shallow and deep taxa complicates any simple interpretation. Our analyses suggest the scales of dispersal and connectivity for reserve design in the deep sea might be comparable to or slightly larger than those in shallow water. Deep‐sea reserve design will need to consider the enormous variety of taxa, life histories, hydrodynamics, spatial configuration of habitats and patterns of species distributions. The many caveats of our analyses provide a strong impetus for substantial future efforts to assess connectivity of deep‐sea species from a variety of habitats, taxonomic groups and depth zones.
Receptors rather than signals change in expression in four physiological regulatory networks during evolutionary divergence in threespine stickleback
Authors: Di Poi Carole , Bélanger Dominic , Amyot Marc , Rogers Sean , Aubin‐Horth Nadia
Published: May 2016

The molecular mechanisms underlying behavioural evolution following colonization of novel environments are largely unknown. Molecules that interact to control equilibrium within an organism form physiological regulatory networks. It is essential to determine whether particular components of physiological regulatory networks evolve or if the network as a whole is affected in populations diverging in behavioural responses, as this may affect the nature, amplitude and number of impacted traits. We studied the regulation of four physiological regulatory networks in freshwater and marine populations of threespine stickleback raised in a common environment, which were previously characterized as showing evolutionary divergence in behaviour and stress reactivity. We measured nineteen components of these networks (ligands and receptors) usingmRNAand monoamine levels in the brain, pituitary and interrenal gland, as well as hormone levels. Freshwater fish showed higher expression in the brain of adrenergic (adrb2a), serotonergic (htr2a) and dopaminergic (DRD2) receptors, but lower expression of the htr2b receptor. Freshwater fish also showed higher expression of the mc2r receptor of the glucocorticoid axis in the interrenals. Collectively, our results suggest that the inheritance of the regulation of these networks may be implicated in the evolution of behaviour and stress reactivity in association with population divergence. Our results also suggest that evolutionary change in freshwater threespine stickleback may be more associated with the expression of specific receptors rather than with global changes of all the measured constituents of the physiological regulatory networks.
Applying landscape genetics to the microbial world
Authors: Dudaniec Rachael Y. , Tesson Sylvie V. M.
Published: May 2016

Landscape genetics, which explicitly quantifies landscape effects on gene flow and adaptation, has largely focused on macroorganisms, with little attention given to microorganisms. This is despite overwhelming evidence that microorganisms exhibit spatial genetic structuring in relation to environmental variables. The increasing accessibility of genomic data has opened up the opportunity for landscape genetics to embrace the world of microorganisms, which may be thought of as ‘the invisible regulators’ of the macroecological world. Recent developments in bioinformatics and increased data accessibility have accelerated our ability to identify microbial taxa and characterize their genetic diversity. However, the influence of the landscape matrix and dynamic environmental factors on microorganism genetic dispersal and adaptation has been little explored. Also, because many microorganisms coinhabit or codisperse with macroorganisms, landscape genomic approaches may improve insights into how micro‐ and macroorganisms reciprocally interact to create spatial genetic structure. Conducting landscape genetic analyses on microorganisms requires that we accommodate shifts in spatial and temporal scales, presenting new conceptual and methodological challenges not yet explored in ‘macro’‐landscape genetics. We argue that there is much value to be gained for microbial ecologists from embracing landscape genetic approaches. We provide a case for integrating landscape genetic methods into microecological studies and discuss specific considerations associated with the novel challenges this brings. We anticipate that microorganism landscape genetic studies will provide new insights into both micro‐ and macroecological processes and expand our knowledge of species’ distributions, adaptive mechanisms and species’ interactions in changing environments.
Unusually limited pollen dispersal and connectivity of Pedunculate oak (Quercus robur) refugial populations at the species' southern range margin
Authors: Moracho E. , Moreno G. , Jordano P. , Hampe A.
Published: May 2016

Low‐latitudinal range margins of temperate and boreal plant species typically consist of scattered populations that persist locally in microrefugia. It remains poorly understood how their refugial habitats affect patterns of gene flow and connectivity, key components for their long‐term viability and evolution. We examine landscape‐scale patterns of historical and contemporary gene flow in refugial populations of the widespread European forest tree Pedunculate oak (Quercus robur) at the species' southwestern range margin. We sampled all adult trees (n = 135) growing in a 20 km long valley and genotyped 724 acorns from 72 mother trees at 17 microsatellite loci. The ten oak stands that we identified were highly differentiated and formed four distinct genetic clusters, despite sporadic historical dispersal being detectable. By far most contemporary pollination occurred within stands, either between local mates (85.6%) or through selfing (6.8%). Pollen exchange between stands (2.6%) was remarkably rare given their relative proximity and was complemented by long‐distance pollen immigration (4.4%) and hybridization with the locally abundant Quercus pyrenaica (0.6%). The frequency of between‐stand mating events decreased with increasing size and spatial isolation of stands. Overall, our results reveal outstandingly little long‐distance gene flow for a wind‐pollinated tree species. We argue that the distinct landscape characteristics of oaks' refugial habitats, with a combination of a rugged topography, dense vegetation and humid microclimate, are likely to increase plant survival but to hamper effective long‐distance pollen dispersal. Moreover, local mating might be favoured by high tree compatibility resulting from genetic purging in these long‐term relict populations.
The magic dance of the alarmones (p)ppGpp
Authors: Steinchen Wieland , Bange Gert
Published: May 2016

The alarmones (p)ppGpp are important second messengers that orchestrate pleiotropic adaptations of bacteria and plant chloroplasts in response to starvation and stress. Here, we review our structural and mechanistic knowledge on (p)ppGpp metabolism including their synthesis, degradation and interconversion by a highly diverse set of enzymes. Increasing structural information shows how (p)ppGpp interacts with an incredibly diverse set of different targets that are essential for replication, transcription, translation, ribosome assembly and metabolism. This raises the question how the chemically rather simple (p)ppGpp is able to interact with these different targets? Structural analysis shows that the diversity of (p)ppGpp interaction with cellular targets critically relies on the conformational flexibility of the 3′ and 5′ phosphate moieties allowing alarmones to efficiently modulate the activity of target structures in a broad concentration range. Current approaches in the design of (p)ppGpp‐analogs as future antibiotics might be aided by the comprehension of conformational flexibility exhibited by the magic dancers (p)ppGpp.
Cytoskeleton reorganization/disorganization is a key feature of induced inaccessibility for defence to successive pathogen attacks
Authors: Moral Juan , Montilla‐Bascón Gracia , Canales Francisco J. , Rubiales Diego , Prats Elena
Published: May 2016

In this work, we investigated the involvement of the long‐term dynamics of cytoskeletal reorganization on the induced inaccessibility phenomenon by which cells that successfully defend against a previous fungal attack become highly resistant to subsequent attacks. This was performed on pea through double inoculation experiments using inappropriate (Blumeria graminis f. sp. avenae, Bga) and appropriate (Erysiphe pisi, Ep) powdery mildew fungi. Pea leaves previously inoculated with Bga showed a significant reduction of later Ep infection relative to leaves inoculated only with Ep, indicating that cells had developed induced inaccessibility. This reduction in Ep infection was higher when the time interval between Bga and Ep inoculation ranged between 18 and 24 h, although increased penetration resistance in co‐infected cells was observed even with time intervals of 24 days between inoculations. Interestingly, this increase in resistance to Ep following successful defence to the inappropriate Bga was associated with an increase in actin microfilament density that reached a maximum at 18–24 h after Bga inoculation and very slowly decreased afterwards. The putative role of cytoskeleton reorganization/disorganization leading to inaccessibility is supported by the suppression of the induced resistance mediated by specific actin (cytochalasin D, latrunculin B) or general protein (cycloheximide) inhibitors.
Motor neuron disease with saccadic abnormalities similar to progressive supranuclear palsy
Authors: Tokushige Shin‐ichi , Terao Yasuo , Matsuda Shun‐ichi , Inomata‐Terada Satomi , Shimizu Takahiro , Tanaka Nobuyuki , Hamada Masashi , Yugeta Akihiro , Hanajima Ritsuko , Mori Harushi , Tsuji Shoji , Ugawa Yoshikazu
Published: May 2016

Background

In recent years, a variety of clinical types of amyotrophic lateral sclerosis have come to be recognized. As some patients present with oculomotor abnormalities both clinically and pathologically, the progressive supranuclear palsy variant of amyotrophic lateral sclerosis has been proposed.

Aim

To describe atypical cases of motor neuron disease with abnormal extraocular movements mimicking progressive supranuclear palsy.

Methods

We present three motor neuron disease patients with slow saccades, who were aged 57, 63 and 62 years. Neurological examinations found vertical gaze palsy in two patients. The two patients who presented extrapyramidal signs were regarded as motor neuron disease with parkinsonism, whereas the other was diagnosed with amyotrophic lateral sclerosis. Their saccades were investigated by visually‐guided saccade and memory‐guided saccade tasks, and were compared with those of 14 age‐matched normal participants (60.3 ± 1.9 years).

Results

In all these patients, the visually‐guided saccade latencies were significantly prolonged compared with normal participants, whereas the memory‐guided saccade latencies were not. The velocity and amplitude of saccades of the patients were significantly reduced in visually‐guided saccade and memory‐guided saccade in comparison with normal participants.

Conclusion

The patterns of saccadic abnormalities in the patients were similar to those of progressive supranuclear palsy patients, suggesting that some patients with motor neuron disease show saccade abnormalities similar to those of progressive supranuclear palsy patients from the clinical and physiological perspective. Motor neuron disease with slow saccades and parkinsonism, as reported here, suggest the existence of progressive supranuclear palsy‐variant amyotrophic lateral sclerosis.
Early onset of cardiomyopathy and intellectual disability in a girl with Danon disease associated with a de novo novel mutation of the LAMP2 gene
Authors: Sugie Kazuma , Yoshizawa Hiroyuki , Onoue Kenji , Nakanishi Yoko , Eura Nobuyuki , Ogawa Megumu , Nakano Tomoya , Sakaguchi Yasuhiro , Hayashi Yukiko K. , Kishimoto Toshifumi , Shima Midori , Saito Yoshihiko , Nishino Ichizo , Ueno Satoshi
Published: May 2016

Danon disease, primary lysosome‐associated membrane protein‐2 (LAMP‐2) deficiency, is characterized clinically by cardiomyopathy, myopathy and intellectual disability in boys. Because Danon disease is inherited in an X‐linked dominant fashion, males are more severely affected than females, who usually have only cardiomyopathy without myopathy or intellectual disability; moreover, the onset of symptoms in females is usually in adulthood. We describe a girl with Danon disease who presented with hypertrophic cardiomyopathy and Wolff‐Parkinson‐White (WPW) syndrome at 12 years of age. Subsequently, she showed signs of mild learning disability and intellectual disability on psychological examinations. She had a de novo novel mutation in the LAMP‐2 gene and harbored an identical c.749C > A (p.Ser250X) variant, resulting in a stop codon in exon 6. She showed decreased, but not completely absent LAMP‐2 expression on immunohistochemical and Western blot analyses of a skeletal muscle biopsy specimen, which has been suggested to be caused by a 50% reduction in LAMP‐2 expression (LAMP‐2 haploinsufficiency) in female patients with Danon disease caused by a heterozygous null mutation. To our knowledge, our patient is one of the youngest female patients to have been given a diagnosis of Danon disease. In addition, this is the first documented case in a girl that was clearly associated with intellectual disability, which is very rare in females with Danon disease. Our findings suggest that studies of female patients with Danon disease can extend our understanding of the clinical features of this rare disease.
Remifentanil and propofol undergo separation and layering when mixed in the same syringe for total intravenous anesthesia
Authors: O'Connor Sean , Zhang Yan Ling , Christians Uwe , Morrison John E. , Friesen Robert H.
Published: May 2016

Background

Propofol and remifentanil can be combined to deliver total intravenous anesthesia (TIVA). Propofol and remifentanil are sometimes mixed in the same syringe. Since remifentanil is a solution and propofol is an emulsion, we hypothesized that they would separate over time when mixed in the same syringe.

Methods

Nine 60‐ml polypropylene syringes were prepared as follows: Group A: 1.25 ml of remifentanil solution (1 mg·ml−1) was added to 48.75 ml of propofol (10 mg·ml−1) in three syringes. Group B: 2.5 ml of remifentanil (1 mg·ml−1) was added to 47.5 ml of propofol (10 mg·ml−1) in three syringes. Group C: 5 ml of remifentanil (1 mg·ml−1) was added to 45 ml of propofol (10 mg·ml−1) in three syringes. The remifentanil lyophilized powder was reconstituted with sterile water and added to the propofol by injection through the port on the bottom of the syringe. The syringe was then inverted five times in succession to mix the drugs. The syringes were mounted in an upright vertical position (plunger on top, port on bottom) with wire on a pegboard. Samples of the mixture were taken from the bottom port (via a 3‐way stopcock) and from the top of the syringe (via a stopcock on an 18‐gauge needle placed 5 mm through the plunger) at the following time intervals (min) from baseline: T0, T10, T30, T60, T120, T180, T240, T300. Remifentanil and propofol were quantified using specific and validated HPLC/MS/MS assays with automated online sample preparation.

Results

Concentrations of remifentanil were significantly greater at the top than the bottom of the syringes in groups A and B. Concentrations of propofol were significantly greater at the bottom than the top of the syringes in all groups.

Conclusion

Our data indicate that remifentanil solution and propofol emulsion are immiscible: remifentanil separates from propofol and rises to the top. Thus, concentrations of remifentanil and propofol delivered to patients from the same syringe during TIVA are not those expected and cannot be reliable. Remifentanil and propofol should be administered in separate syringes when used in combination for TIVA.
The antagonistic regulation of abscisic acid‐inhibited root growth by brassinosteroids is partially mediated via direct suppression of ABSCISIC ACID INSENSITIVE 5 expression by BRASSINAZOLE RESISTANT 1
Authors: Yang Xiaorui , Bai Yang , Shang Jianxiu , Xin Ruijiao , Tang Wenqiang
Published: May 2016

Brassinosteroids (BRs) and abscisic acid (ABA) are plant hormones that antagonistically regulate many aspects of plant growth and development; however, the mechanisms that regulate the crosstalk of these two hormones are still not well understood. BRs regulate plant growth and development by activating BRASSINAZOLE RESISTANT 1 (BZR1) family transcription factors. Here we show that the crosstalk between BRs and ABA signalling is partially mediated by BZR1 regulated gene expression. bzr1‐1D is a dominant mutant with enhanced BR signalling; our results showed that bzr1‐1D mutant is less sensitive to ABA‐inhibited primary root growth. By RNA sequencing, a subset of BZR1 regulated ABA‐responsive root genes were identified. Of these genes, the expression of a major ABA signalling component ABA INSENSITIVE 5 (ABI5) was found to be suppressed by BR and by BZR1. Additional evidences showed that BZR1 could bind strongly with several G‐box cis‐elements in the promoter of ABI5, suppress the expression of ABI5 and make plants less sensitive to ABA. Our study demonstrated that ABI5 is a direct target gene of BZR1, and modulating the expression of ABI5 by BZR1 plays important roles in regulating the crosstalk between the BR and ABA signalling pathways.
UpStart Parent Survey—Prenatal: A New Tool for Evaluating Prenatal Education Programs
Authors: Benzies Karen M. , Barker Leslie , Churchill Jocelyn , Smith Jennifer , Horn Sarah
Published: May 2016

Objectives

To evaluate a new prenatal education program evaluation tool, the UpStart Parent Survey – Prenatal, in terms of: (a) reliability and validity; (b) sensitivity to change over time; (c) whether results differed for mothers versus fathers; and (d) whether results differed when using an electronic tablet‐computer versus a paper survey.

Design and Sample

Psychometric study. Participants were 277 expectant mothers (n = 161) and fathers (n = 106) enrolled in Childbirth Essentials, a 6‐week prenatal education program.

Measures

The UpStart Parent Survey – Prenatal is a retrospective pretest/posttest survey with three scales: Parenting Knowledge, Parenting Experience, and Program Satisfaction, and three open‐ended questions.

Results

The UpStart Parent Survey – Prenatal is sensitive to change and demonstrated significant positive differences in parenting knowledge and parenting experience. There was no difference in results whether the survey was completed by mothers or fathers. Results were similar whether paper or electronic formats were used. The survey was easy to complete.

Conclusion

The UpStart Parent Survey – Prenatal holds promise as a reliable and valid evaluation tool to capture outcomes of brief prenatal education programs that target the general population of expectant parents.
Confidence Intervals and Regions for the Lasso by Using Stochastic Variational Inequality Techniques in Optimization
Authors: Lu Shu , Liu Yufeng , Yin Liang , Zhang Kai
Published: May 2016

Sparse regression techniques have been popular in recent years because of their ability in handling high dimensional data with built-in variable selection. The lasso is perhaps one of the most well-known examples. Despite intensive work in this direction, how to provide valid inference for sparse regularized methods remains a challenging statistical problem. We take a unique point of view of this problem and propose to make use of stochastic variational inequality techniques in optimization to derive confidence intervals and regions for the lasso. Some theoretical properties of the procedure are obtained. Both simulated and real data examples are used to demonstrate the performance of the method.
The Climate Policy Hold‐Up: Green Technologies, Intellectual Property Rights, and the Abatement Incentives of International Agreements
Authors: Goeschl Timo , Perino Grischa
Published: May 2016

The success of global climate policies over the coming decades depends on the diffusion of “green” technologies. Using a simple model, we highlight a conflict between international environmental agreements (IEAs) on emissions reductions and international systems of intellectual property rights (IPRs) on abatement technologies. When IPRs are strong and global, IEA signatories anticipate rent extraction by innovators. This hold‐up effect reduces abatement, potentially to levels below those of non‐signatories, and it reduces the number of signatories to self‐enforcing IEAs. We explore policy options that respect existing property rights, but avoid the strategic interaction between signatories to an IEA and innovators.
How Productive Is Workplace Health and Safety?
Authors: Buhai I. Sebastian , Cottini Elena , Westergaard‐Nielsen Niels
Published: May 2016

In this paper, we investigate the causal impact of workplace health and safety practices on firm performance, using Danish longitudinal matched employer–employee data merged with unique cross‐sectional representative firm survey data on work environment conditions. We estimate standard production functions, augmented with workplace environment indicators, addressing both time‐invariant and time‐varying potentially relevant unobservables in the production process. We find positive and large productivity effects of improved physical dimensions of the health and safety environment, specifically, “internal climate” and “monotonous repetitive work”.
Aminooxy‐naphthylpropionic acid and its derivatives are inhibitors of auxin biosynthesis targeting l‐tryptophan aminotransferase: structure–activity relationships
Authors: Narukawa‐Nara Megumi , Nakamura Ayako , Kikuzato Ko , Kakei Yusuke , Sato Akiko , Mitani Yuka , Yamasaki‐Kokudo Yumiko , Ishii Takahiro , Hayashi Ken‐ichiro , Asami Tadao , Ogura Takehiko , Yoshida Shigeo , Fujioka Shozo , Kamakura Takashi , Kawatsu Tsutomu , Tachikawa Masanori , Soeno Kazuo , Shimada Yukihisa
Published: May 2016

We previously reported l‐α‐aminooxy‐phenylpropionic acid (AOPP) to be an inhibitor of auxin biosynthesis, but its precise molecular target was not identified. In this study we found that AOPP targets TRYPTOPHAN AMINOTRANSFERASE of ARABIDOPSIS 1 (TAA1). We then synthesized 14 novel compounds derived from AOPP to study the structure–activity relationships of TAA1 inhibitors in vitro. The aminooxy and carboxy groups of the compounds were essential for inhibition of TAA1 in vitro. Docking simulation analysis revealed that the inhibitory activity of the compounds was correlated with their binding energy with TAA1. These active compounds reduced the endogenous indole‐3‐acetic acid (IAA) content upon application to Arabidopsis seedlings. Among the compounds, we selected 2‐(aminooxy)‐3‐(naphthalen‐2‐yl)propanoic acid (KOK1169/AONP) and analyzed its activities in vitro and in vivo. Arabidopsis seedlings treated with KOK1169 showed typical auxin‐deficient phenotypes, which were reversed by exogenous IAA. In vitro and in vivo experiments indicated that KOK1169 is more specific for TAA1 than other enzymes, such as phenylalanine ammonia‐lyase. We further tested 41 novel compounds with aminooxy and carboxy groups to which we added protection groups to increase their calculated hydrophobicity. Most of these compounds decreased the endogenous auxin level to a greater degree than the original compounds, and resulted in a maximum reduction of about 90% in the endogenous IAA level in Arabidopsis seedlings. We conclude that the newly developed compounds constitute a class of inhibitors of TAA1. We designated them ‘pyruvamine’.
Safety evaluation of combination doxorubicin and toceranib phosphate (Palladia®) in tumour bearing dogs: a phase I dose‐finding study
Authors: Pellin M. A. , Wouda R. M. , Robinson K. , Tsimbas K. , Kurzman I. D. , Biller B. J. , Vail D. M.
Published: May 2016

Combination chemotherapy holds promise for improving outcomes in malignancy when compared with single‐agent approaches. Care must be taken to avoid overlapping toxicity and to utilize agents with differing mechanisms of action. A phase I dose‐finding trial was performed to determine the maximally tolerated dose (MTD) of a concurrent toceranib and doxorubicin (DOX) combination protocol where toceranib dose was maintained at or near 2.75 mg kg−1 by mouth every other day (PO EOD) while escalating DOX dosage. The dose‐limiting toxicity was found to be neutropenia and the MTD of the combination was determined to be 25 mg m−2 of DOX q 21 days given concurrently with toceranib 2.75 mg kg−1 PO EOD. This combination was well tolerated with no excessive gastrointestinal toxicity nor novel adverse events (AEs) noted. Anti‐tumour activity was observed in the majority of cases. This combination warrants further investigation in the context of phase II/III clinical trials to characterize efficacy and long‐term AE profiles.
SWEDENBORG AND THE PLURALITY OF WORLDS: ASTROTHEOLOGY IN THE EIGHTEENTH CENTURY
Authors: Dunér David
Published: May 2016

The possible existence of extraterrestrial life led in the eighteenth century to a heated debate on the unique status of the human being and of Christianity. One of those who discussed the new scientific worldview and its implications for theology was the Swedish natural philosopher and theologian Emanuel Swedenborg. This article discusses Swedenborg's astrotheological transformation, his use of theological arguments in his early cosmology, and his cosmogony that later on ended up in his use of contemporary natural philosophy in his theology, especially concerning the question of the plurality of worlds. I will first sketch the astrotheology found in his natural philosophical works, and then turn to the astrotheology of his later spiritual teachings. In Swedenborg's works we find teleological arguments and a stress on the universality of the divine creation and Christianity, as well as anthropomorphic descriptions of extraterrestrial life. By reconciling contemporary astronomical ideas, among others the concept of the plurality of worlds, with Christian dogmas, Swedenborg refuted deistic conclusions that Jesus was merely a mortal, while at the same time keeping his belief in the modern astronomical worldview.
Antibody Persistence in Young Children 5 Years after Vaccination with a Combined Haemophilus influenzae Type b-Neisseria meningitidis Serogroup C Conjugate Vaccine Coadministered with Diphtheria-Tetanus-Acellular Pertussis-Based and Pneumococcal Conjugate Vaccines
Authors: Tejedor Juan Carlos , Brzostek Jerzy , Konior Ryszard , Grunert Detlef , Kolhe Devayani , Baine Yaela , Van Der Wielen Marie
Published: May 2016

We evaluated antibody persistence in children up to 5 years after administration of a combined
Haemophilus influenzae
type b (Hib)-
Neisseria meningitidis
serogroup C (MenC)-tetanus toxoid (TT) conjugate vaccine coadministered with a pneumococcal conjugate vaccine. This is the follow-up study of a randomized trial (ClinicalTrials.gov registration no. NCT00334334/00463437) in which healthy children were vaccinated (primary vaccinations at 2, 4, and 6 months of age and booster vaccination at 11 to 18 months of age) with Hib-MenC-TT or a control MenC conjugate vaccine, coadministered with diphtheria-tetanus-acellular pertussis (DTPa)-based combination vaccines (DTPa/Hib for control groups) and a pneumococcal conjugate vaccine (10-valent pneumococcal nontypeable
H. influenzae
protein D conjugate vaccine [PHiD-CV] or 7-valent cross-reacting material 197 [CRM
197
] conjugate vaccine [7vCRM]). MenC antibody titers were measured with a serum bactericidal antibody (SBA) assay using rabbit complement (i.e., rabbit SBA [rSBA]), and antibodies against Hib polyribosylribitol phosphate (PRP) were measured with an enzyme-linked immunosorbent assay. Antibody persistence up to 5 years after booster vaccination is reported for 530 children ∼6 years of age. The percentages of children with seroprotective rSBA-MenC titers were between 24.2% and 40.1% in all groups approximately 5 years after booster vaccination. More than 98.5% of children in each group retained seroprotective anti-PRP concentrations. No vaccine-related serious adverse events and no events related to a lack of vaccine efficacy were reported. Approximately 5 years after booster vaccination, the majority of children retained seroprotective anti-PRP antibody concentrations. The percentage of children retaining seroprotective rSBA-MenC titers was low (≤40%), suggesting that a significant proportion of children may be unprotected against MenC disease. (This study has been registered at ClinicalTrials.gov under registration no. NCT00891176.)
Sequencing Analysis of NS3/4A, NS5A, and NS5B Genes from Patients Infected with Hepatitis C Virus Genotypes 5 and 6
Authors: Ku Karin S. , Chodavarapu Ramakrishna K. , Martin Ross , Miller Michael D. , Mo Hongmei , Svarovskaia Evguenia S.
Published: May 2016

Direct-acting antivirals (DAAs) with activity against multiple genotypes of the hepatitis C virus (HCV) were recently developed and approved for standard-of-care treatment. However, sequencing assays to support HCV genotype 5 and 6 analysis are not widely available. Here, we describe the development of a sequencing assay for the NS3/4A, NS5A, and NS5B genes from HCV genotype 5 and 6 patient isolates. Genotype- and subtype-specific primers were designed to target NS3/4A, NS5A, and NS5B for cDNA synthesis and nested PCR amplification. Amplification was successfully performed for a panel of 32 plasma samples from HCV-infected genotype 5 and 6 patients with sequencing data obtained for all attempted samples. LiPA 2.0 (Versant HCV genotype 2.0) is a reverse hybridization line probe assay that is commonly used for genotyping HCV-infected patients enrolled in clinical studies. Using NS3/4A, NS5A, and NS5B consensus sequences, HCV subtypes were determined that were not available for the initial LiPA 2.0 result for genotype 6 samples. Samples amplified here included the following HCV subtypes: 5a, 6a, 6e, 6f, 6j, 6i, 6l, 6n, 6o, and 6p. The sequencing data generated allowed for the determination of the presence of variants at amino acid positions previously characterized as associated with resistance to DAAs. The simple and robust sequencing assay for genotypes 5 and 6 presented here may lead to a better understanding of HCV genetic diversity and prevalence of resistance-associated variants.
Lassa Virus Cell Entry via Dystroglycan Involves an Unusual Pathway of Macropinocytosis
Authors: Oppliger Joel , Torriani Giulia , Herrador Antonio , Kunz Stefan
Published: May 2016

The pathogenic Old World arenavirus Lassa virus (LASV) causes a severe hemorrhagic fever with a high rate of mortality in humans. Several LASV receptors, including dystroglycan (DG), TAM receptor tyrosine kinases, and C-type lectins, have been identified, suggesting complex receptor use. Upon receptor binding, LASV enters the host cell via an unknown clathrin- and dynamin-independent pathway that delivers the virus to late endosomes, where fusion occurs. Here we investigated the mechanisms underlying LASV endocytosis in human cells in the context of productive arenavirus infection, using recombinant lymphocytic choriomeningitis virus (rLCMV) expressing the LASV glycoprotein (rLCMV-LASVGP). We found that rLCMV-LASVGP entered human epithelial cells via DG using a macropinocytosis-related pathway independently of alternative receptors. Dystroglycan-mediated entry of rLCMV-LASVGP required sodium hydrogen exchangers, actin, and the GTPase Cdc42 and its downstream targets, p21-activating kinase-1 (PAK1) and Wiskott-Aldrich syndrome protein (N-Wasp). Unlike other viruses that enter cells via macropinocytosis, rLCMV-LASVGP entry did not induce overt changes in cellular morphology and hardly affected actin dynamics or fluid uptake. Screening of kinase inhibitors identified protein kinase C, phosphoinositide 3-kinase, and the receptor tyrosine kinase human hepatocyte growth factor receptor (HGFR) to be regulators of rLCMV-LASVGP entry. The HGFR inhibitor EMD 1214063, a candidate anticancer drug, showed antiviral activity against rLCMV-LASVGP at the level of entry. When combined with ribavirin, which is currently used to treat human arenavirus infection, EMD 1214063 showed additive antiviral effects. In sum, our study reveals that DG can link LASV to an unusual pathway of macropinocytosis that causes only minimal perturbation of the host cell and identifies cellular kinases to be possible novel targets for therapeutic intervention.

IMPORTANCE
Lassa virus (LASV) causes several hundred thousand infections per year in Western Africa, with the mortality rate among hospitalized patients being high. The current lack of a vaccine and the limited therapeutic options at hand make the development of new drugs against LASV a high priority. In the present study, we uncover that LASV entry into human cells via its major receptor, dystroglycan, involves an unusual pathway of macropinocytosis and define a set of cellular factors implicated in the regulation of LASV entry. A screen of kinase inhibitors revealed HGFR to be a possible candidate target for antiviral drugs against LASV. An HGFR candidate inhibitor currently being evaluated for cancer treatment showed potent antiviral activity and additive drug effects with ribavirin, which is used in the clinic to treat human LASV infection. In sum, our study reveals novel fundamental aspects of the LASV-host cell interaction and highlights a possible candidate drug target for therapeutic intervention.