bluuebunny/crossref_metadata_2025 huggingface dataset:
Knowledge sharing between general and pediatric emergency departments: connections, barriers, and opportunities
Authors: Crockett Leah K. , Leggett Carly , Curran Janet A. , Knisley Lisa , Brockman Gwenyth , Scott Shannon D. , Hartling Lisa , Jabbour Mona , Klassen Terry P.
Published: February 2018

Objective

Translating Emergency Knowledge for Kids (TREKK) is a national network aimed at improving emergency care for children by increasing collaborations and knowledge sharing between general and pediatric emergency departments (EDs). This study aimed to determine patterns of knowledge sharing within the network and to identify connections, barriers, and opportunities to obtaining pediatric information and training.

Methods

We conducted 22 semi-structured interviews with health care professionals working in general EDs, purposefully sampled to represent connected and disconnected sites, based on two previous internal quantitative social network analyses (SNA). Data were analyzed by two independent reviewers.

Results

Participants included physicians (59%) and nurses (41%) from 18 general EDs in urban (68%) and rural/remote (32%) Canada. Health care professionals sought information both formally and informally, by using guidelines, talking to colleagues, and attending pediatric related training sessions. Network structure and processes were found to increase connections, support practice change, and promote standards of care. Participants identified personal, organizational and system level barriers to information and skill acquisition, including resources and personal costs, geography, dissemination, and time. Providing easy access to information at the point of care was promoted through enhancing content visibility and by embedding resources into local systems. There remains a need to share successful methods of local dissemination and implementation across the network, and to leverage local professional champions such as clinical nurse liaisons.

Conclusions

These findings reinforce the critical role of ongoing network evaluation to improve the design and delivery of knowledge mobilization initiatives.
Primary Care Medical Practices: Are Community Health Care Providers Ready for Disasters?
Authors: Peters Rachel M. , Hipper Thomas J. , Chernak Esther D.
Published: February 2018

Objective

This study seeks to determine the capacity of community primary care practices to meet the needs of patients during public health emergencies and to identify the barriers and resources necessary to participate in a coordinated response with public safety agencies.

Methods

The self-administered web-based survey was distributed in January 2014 via e-mail to primary care providers in Pennsylvania using the listservs of several professional societies.

Results

A total of 179 primary care providers participated in the survey. In total, 38% had practice continuity of operations plan in place and 26% reported that they had a plan for patient surge in the outpatient setting. Thirty percent reported that they were registered on the state Health Alert Network and 41% said they were able to communicate with patients during disasters. Only 8% of providers reported that they believed that their patients with special health care needs were prepared for a disaster, although over two-thirds of responding practices felt they could assist these patients with disaster preparedness. Providers indicated that more information regarding government agency plans and community resources, patient education materials, and more time to devote to counseling during patient encounters would improve their ability to prepare their patients with special health care needs for disasters. Providers also reported that they would benefit from partnerships to help the practice during emergencies and communications technology to reach large numbers of patients quickly.

Conclusions

Community-based primary care practices can be useful partners during public health emergencies. Efforts to promote continuity of operations planning, improved coordination with government and community partners, as well as preparedness for patients with special health care needs, would augment their capabilities and contribute to community resilience. (Disaster Med Public Health Preparedness. 2019;13:128–132).
Creation of a Collaborative Disaster Preparedness Video for Daycare Providers: Use of the Delphi Model for the Creation of a Comprehensive Disaster Preparedness Video for Daycare Providers
Authors: Mar Pamela , Spears Robert , Reeb Jeffrey , Thompson Sarah B. , Myers Paul , Burke Rita V.
Published: February 2018

Objective

Eight million American children under the age of 5 attend daycare and more than another 50 million American children are in school or daycare settings. Emergency planning requirements for daycare licensing vary by state. Expert opinions were used to create a disaster preparedness video designed for daycare providers to cover a broad spectrum of scenarios.

Methods

Various stakeholders (17) devised the outline for an educational pre-disaster video for child daycare providers using the Delphi technique. Fleiss κ values were obtained for consensus data. A 20-minute video was created, addressing the physical, psychological, and legal needs of children during and after a disaster. Viewers completed an anonymous survey to evaluate topic comprehension.

Results

A consensus was attempted on all topics, ranging from elements for inclusion to presentation format. The Fleiss κ value of 0.07 was obtained. Fifty-seven of the total 168 video viewers completed the 10-question survey, with comprehension scores ranging from 72% to 100%.

Conclusion

Evaluation of caregivers that viewed our video supports understanding of video contents. Ultimately, the technique used to create and disseminate the resources may serve as a template for others providing pre-disaster planning education. (Disaster Med Public Health Preparedness. 2019;13:123–127)
Multiple glacial advances in the Rangitata Valley, South Island, New Zealand, imply roles for Southern Hemisphere westerlies and summer insolation in MIS 3 glacial advances
Authors: Shulmeister James , Thackray Glenn D. , Rittenour Tammy M. , Hyatt Olivia M.
Published: February 2018

Stratigraphic evidence and extensive optically stimulated luminescence (OSL) geochronology from an 18-km-long reach of the middle Rangitata Valley, South Island, New Zealand, provide evidence for at least six distinct glacial advances during the last glacial cycle. These include four well-constrained Marine Oxygen Isotope Stage (MIS) 3 and 2 advances at ca. 38 ka, ca. 27 ka, ca. 21 ka and at 18 ka, as well as less well-constrained advances in MIS 4 and/or early MIS 3. Ice occupied a farther downvalley reach of the Rangitata from 38 ka to after 18 ka, indicating that near-full glacial conditions persisted for most of the last 20 ka of the last glaciation, though the glacier still fluctuated significantly, as reflected by the numerous distinguishable advances. Global or regional cooling alone cannot explain the persistence of near-maximum glacial conditions for this extended period, nor can it explain the occurrence of the largest advances ca. 32 ka. Instead, we invoke the northward expansion of the westerlies during MIS 3 as the cause for the early widespread glaciation, wherein enhanced westerly flow under moderate cooling maximised glacial extents. Local insolation favoured extended MIS 3 glaciation until ca. 32 ka. Increasing summer insolation gradually reduced glacial extents after ca. 28 ka.
Allowing for the structure of a designed experiment when estimating and testing trait correlations
Authors: Piepho Hans-Peter
Published: February 2018

Crop scientists occasionally compute sample correlations between traits based on observed data from designed experiments, and this is often accompanied by significance tests of the null hypothesis that traits are uncorrelated. This simple approach does not account for effects due to the randomization layout and treatment structure of the experiments and hence statistical inference based on standard procedures is not appropriate. The present paper describes how valid inferences accounting for all relevant effects can be obtained using bivariate mixed linear model procedures. A salient feature of the approach is that the bivariate model is commensurate with the model used for univariate analysis of individual traits and allows bivariate correlations to be computed at the level of effects. Heterogeneity of correlations between effects can be assessed by likelihood ratio tests or by graphical inspection of bivariate scatter plots of effect estimates. if heterogeneity is found to be substantial, it is crucial to focus on the correlation of effects, and usually, the main interest will be in the treatment effects. If heterogeneity is judged to be negligible, the marginal correlation can be estimated from the bivariate model for an overall assessment of association. The proposed methods are illustrated using four examples. Hints are given to alternative routes of analysis accounting for all treatment and design effects such as regression with groups and analysis of covariance.
Prediction of sheep milk chemical composition using milk yield, pH, electrical conductivity and refractive index
Authors: Gelasakis Athanasios I , Giannakou Rebecca , Valergakis Georgios E , Fortomaris Paschalis , Kominakis Antonios , Arsenos Georgios
Published: February 2018

This Research Communication addresses the hypothesis that fat, protein, lactose and total solids content can be predicted using daily milk yield (DMY), pH, electrical conductivity (MEC) and refractive index (RI) of milk as predictors. It also addresses the possibility of these measurements being used for on-farm benchmarking activities towards selecting the highest yielding animals and flocks regarding milk quality traits (MQT). A total of 308 purebred Frizarta ewes were used for the study. From each individual ewe, a composite milk sample was collected. pH, MEC and RI of milk were measured and the samples were assayed for fat, protein, lactose and total solids content, using an automatic infrared milk analyser. The predictive value of DMY, pH, MEC and RI of milk on its MQT was assessed using multiple linear regression analysis. Significant regression equations were produced for all of the studied traits. RI and MEC were significant and reliable predictors for all studied MQT, whereas DMY was a significant predictor for most MQT with the exception of protein content. pH was a marginally significant predictor for some of the MQTs at the initial development of the equations but proved unreliable after bootstraping. Using these equations a number of ewes varying from 75 (for fat) to 97 (for protein) out of the 100 highest MQT yielders were correctly predicted, whereas, none of the ewes out of the 100 lowest MQT yielders was mispredicted as a high yielder for protein-, lactose- and total solids- content. Three out of 100 lowest fat-yielders were mispredicted as high fat-yielders. Similar equations can be used for benchmarking activities towards selecting the highest protein-, fat-, lactose- and total solids- yielding animals and flocks in cases where laboratories for MQT analyses are not readily available or the cost of chemical analyses is high. The method can be regarded as a handy tool for the dairy industry to readily assess milk quality at the farm level.
Effect of heat stress on the interaction of Streptococcus uberis with bovine mammary epithelial cells
Authors: Almeida Raúl A. , Kerro-Dego Oudessa , Rius Agustín G.
Published: February 2018

Heat stress (HS) negatively affects milk production and has been associated with decreased immune function, and increased rate of intramammary infections (IMI). Research has shown that HS affects gene expression, cell cycle, and cell metabolism in bovine mammary epithelial cells (BMEC). Since BMEC are an initial target of mastitis pathogens, we studied adherence to and internalisation of S. uberis into HS-BMEC, as well as the effect that this interaction has on host cells by measuring HS-BMEC viability and membrane integrity. Results reported in this Research Communication showed that HS reduced cell viability and induced membrane damage. However, these pathological changes, as well as the rate of adherence and internalisation of S. uberis into BMEC, were augmented when S. uberis was cocultured with HS-BMEC. These results may help to understand the pathogenesis of S. uberis IMI as well as the increased susceptibility of mammary glands to IMI in cows subjected to HS.
Reticulorumen temperature and pH as indicators of the likelihood of reproductive success
Authors: Antanaitis Ramūnas , Juozaitienė Vida , Rutkauskas Arūnas , Televičius Mindaugas , Stasiulevičiūtė Indrė
Published: February 2018

The temperature and pH of the contents of the reticulorumen of cattle can be monitored using sensors placed in a rumen bolus. In this research communication we test the hypothesis that these measurements can be used as a predictor of the likelihood of reproductive success. The study was performed on 20 Lithuanian Black and White dairy cows. The pH and temperature of the contents of their reticulorumens were measured using boluses manufactured for animal care (smaXtec animal care technology®). The data from reticulorumen were recording at 1 to 7 and 7 to 56 d post calving and then post insemination (on average 60 ± 12) until 170 d after calving. The pregnancies were tested with ‘Easy scan’ ultrasound 30–35 d after insemination. Those cows which were pregnant were assigned to the PD+ group (n = 12), and those which were not pregnant – assigned to the PD− group (n = 8). Rumen pH was initially lower in the PD+ group but then higher than in PD−, but temperature did not differ between the groups until after conception. We can conclude that cows with disturbed rumen metabolism (with low rumen pH) are less likely to conceive. The measurements of reticuloruminal pH may be useful as a predictor of the likelihood of reproductive success, but more research is needed before practical application.
Use of contrast-enhanced ultrasonographic examination to evaluate health status of mammary glands of ewes at the end of a lactation period
Authors: Mantziaras George , Vasileiou Natalia GC , Ioannidi Katerina S , Mavrogianni Vasiliki S , Gougoulis Dimitrios A , Fthenakis George C , Petridis Ioannis G , Barbagianni Mariana S
Published: February 2018

This research communication describes the use of contrast-enhanced ultrasonographic examination (CEUS) in mammary glands of ewes for diagnosis of chronic mastitis; this is the first report of the use of this modality in diagnostic imaging of mammary glands of ruminants. For this purpose, a convex transducer was used, with the following settings: frequency: 2·0/4·0 MHz, mechanical index: 0·09, power: 22 dB, scanning depth: 70 mm, and sulphur hexafluoride in microbubbles at a dose of 20 µl as the contrast agent. In four healthy mammary glands (2 ewes), CEUS examination revealed a steady biphasic pattern of contrast agent kinetics characterised by initial uptake within 15–40 s post-injection, at which time intensity peaked with strong enhancement (130–200 AEU) followed by a gradual wash-out phase. In three mammary glands with history of clinical mastitis (2 ewes), the pattern was particularly inconsistent and unclear, with weak enhancement (<100 AEU) (P < 0·01) lasting for a short period. Notwithstanding issues regarding cost and withdrawal period of contrast-agent, this imaging modality may contribute to improved diagnosis of mastitis cases, especially on occasions when abnormalities cannot be easily confirmed by more conventional methods.
The comforting power of Christianity: Genevieve Aupidlak's correspondence with Father A. Thibert (OMI) in the Brandon Sanatorium (1954–1957)
Authors: Laugrand Frédéric B.
Published: February 2018

In 2010, after many interviews by the Qikiqtani Truth Commission (QTC), the Qikiqtani Inuit Association released a report to help the public understand the impact of tuberculosis on the Inuit and thereby promote reconciliation, a goal shared by this study. In this paper, I discuss a corpus of about 28 letters by an Inuit patient named Genevieve Aupidlak who was treated at the Brandon Sanatorium from 1954 to 1957. The letters have been transcribed from old syllabics and translated. Aupidlak's story shows not only the suffering of Inuit in southern sanatoria but also the role of Christianity and that of some missionaries—such as Father Thibert—in supporting Inuit resilience. Aupidlak's letters illustrate how Inuit used their Christian faith to survive and how they ascribed healing power to Christian prayer and confession, in keeping with a much older shamanic pattern.
Developmental competence of cat (Felis domesticus) oocytes and embryos after parthenogenetic stimulation using different methods
Authors: Kochan Joanna , Nowak Agnieszka , Niżański Wojciech , Prochowska Sylwia , Migdał Anna , Młodawska Wiesława , Partyka Agnieszka , Witkowski Maciej
Published: February 2018

The aim of this study was to compare the effects of various activating factors on feline oocytes. The study included activation within the ovary (natural), activation during in vitro maturation (spontaneous activation), chemical activation (ionomycin + 6-DMAP), activation by spermatozoa and injection (ICSI) and mechanical activation (sham ICSI). According to our results, parthenogenetic embryos could emerge at every step of in vitro embryo production (IVP) procedures. After oocyte collection, 6% of parthenogenetic embryos were observed, mainly at the 2–4-blastomere stages. After 24 h of in vitro maturation, parthenogenetic activation was observed in 7% of oocytes. Using ionomycin and 6-DMAP to artificially activate oocytes, 53% of cleaved embryos were obtained. The results after ICSI (54% cleaved embryos) were not significantly different from the results in Group III using chemical activation (53% cleaved embryos). But only after ICSI were blastocysts obtained (5/73.7%) as a result of in vitro culture. Moreover, embryos after ICSI were of the best morphological quality with minor levels of fragmentation evident in the embryos. After sham mechanical activation, ‘sham ICSI’, 8% of cleaved embryos were noted. Therefore, it is advised to maintain a negative control in parallel with each step of IVP techniques, to avoid misleading results. Chemical methods for artificial activation of feline oocytes are the most promising for application to the cloning and production of parthenogenetic embryos for experimental studies.
Myocardial stress perfusion magnetic resonance in children with hypertrophic cardiomyopathy
Authors: Hernandez Lazaro E.
Published: February 2018

Background

Microvascular dysfunction in hypertrophic cardiomyopathy has been associated with poor clinical outcome. Several studies have demonstrated a reduced perfusion reserve proportional to the magnitude of the hypertrophy. We investigated the utility of stress perfusion cardiac MRI to detect microvascular dysfunction in children with hypertrophic cardiomyopathy.

Methods

From January 2016 to January 2017, 13 patients, with a mean age of 15.3 years, with hypertrophic cardiomyopathy underwent regadenoson stress perfusion cardiac MRI (1.5-T Siemens Aera). A single-shot, T1-weighted saturation recovery gradient echo was used for first-pass perfusion in a multiple-slices group, including three short-axis slices and one four-chamber slice. Coronary vasodilatory stress was achieved using bolus injection of regadenoson (lexiscan 0.4 mg/5 ml) and dynamic perfusion during rest and stress performed by administering 0.05 mmol/kg of gadolinium contrast agent (gadoteridol) injected at a rate of 3.5 ml/s, followed by assessment of viability using two-dimensional phase-sensitive inversion recovery of the entire myocardium.

Results

All patients completed protocols with no interruptions. In all, seven patients developed perfusion defects after the administration of regadenoson. Asymmetric septal hypertrophy was the most common pattern of hypertrophic cardiomyopathy (n=4) in those with abnormal perfusion. A total of four patients with perfusion defects had a maximum wall thickness <30 mm. The finding of perfusion defects in areas without late gadolinium enhancement in some of our patients indicates that gadolinium enhancement by itself could underestimate the true extension of microvascular disease. Out of seven patients, five patients with positive stress cardiac MRI have undergone implantable cardioverter defibrillator placement based on current guidelines.

Conclusions

Regadenoson stress cardiac MRI is feasible and clinically valuable in paediatric patients. It is particularly effective in unmasking abnormal myocardial perfusion in the presence of microvascular dysfunction in children with hypertrophic cardiomyopathy.
Aurangzeb and the Nāth Yogīs
Authors: BOUILLIER VÉRONIQUE
Published: February 2018

This article focuses on some representations of Aurangzeb in the Nath Yogi lore; their ambivalence appears as a signifier of the complex relationships the Nath Yogis had with Islam. Aurangzeb figures both as a powerful enemy and as a clumsy devotee, he is depicted more as a symbol than as an historical individual character.
Trainees’ experience of cognitive behavioural therapy training: a mixed methods systematic review
Authors: Jenkins Hannah , Waddington Louise , Thomas Nicola , Hare Dougal Julian
Published: February 2018

Research in the field of cognitive behavioural therapy (CBT) has primarily focused on the acquisition and development of skills and competence. Little is known regarding the experience of training from trainees’ perspectives. This systematic review aimed to review and critique the research conducted on the experience of CBT training. Four electronic databases were searched for published studies reporting on the experience of CBT training. Thirteen articles were selected based on pre-determined inclusion and exclusion criteria and were assessed for quality using the Quality Assessment Tool for Studies with Diverse Designs (QATSDD; Sirriyeh et al., 2012). Due to the lack of consistency in the study designs and outcome measures used, a narrative synthesis of the findings was conducted. Findings were categorized within three themes for synthesis: ‘experience of benefit’, ‘internal processes of engagement’ and ‘external influences on engagement’. Overall, this review was able to draw conclusions regarding the experiences of aspects of CBT training from relatively good quality research. However, the review also highlights the lack of studies exploring specific hypotheses regarding the experience of training.
A Novel Way to Track Magma Flow
Authors: Underwood Emily
Published: February 2018

Sparse seismic data can accurately predict volcanic eruptions.
RETRACTED ARTICLE: A novel microRNA identified in hepatocellular carcinomas is responsive to LEF1 and facilitates proliferation and epithelial-mesenchymal transition via targeting of NFIX
Authors: Hu Yaqi , Guo Xu , Wang Jinxia , Liu Yankun , Gao Huijie , Fan Hongxia , Nong Xiangyang , Yang Xi , Liu Min , Li Shengping , Tang Hua
Published: February 2018

Hepatocellular carcinoma (HCC) is one of the most prevalent cancers. It has been demonstrated that various cellular microRNAs (miRNAs) play an important role in HCC development. Here, we analyzed the miRNA profile in HCC tissues by Solexa sequencing, and we identified a novel microRNA, miR-HCC1, which is upregulated in HCC tissues. Further experiments showed that miR-HCC1 promoted HCC cell proliferation in vivo and in vitro, and migration and invasion resulting from the epithelial-mesenchymal transition (EMT) process. Nuclear factor I/X (NFIX), which inhibited cell proliferation, migration and invasion in HCC cells, was identified as a direct and functional target of miR-HCC1. Furthermore, lymphoid enhancer binding factor 1 (LEF1), a transcription factor, was shown to bind the promoter of miR-HCC1 and activate its expression. Collectively, these results indicate that LEF1-upregulated miR-HCC1 functions as an oncogene through the negative regulation of NFIX expression, which links the LEF1/miR-HCC1/NFIX axis to contribute to cell proliferation, migration and invasion of HCC cells and could provide novel insights into miRNA function and hepatocarcinogenesis and potential biomarkers for HCC.
Efficacy of the highly selective focal adhesion kinase inhibitor BI 853520 in adenocarcinoma xenograft models is linked to a mesenchymal tumor phenotype
Authors: Hirt Ulrich A. , Waizenegger Irene C. , Schweifer Norbert , Haslinger Christian , Gerlach Daniel , Braunger Jürgen , Weyer-Czernilofsky Ulrike , Stadtmüller Heinz , Sapountzis Ioannis , Bader Gerd , Zoephel Andreas , Bister Bojan , Baum Anke , Quant Jens , Kraut Norbert , Garin-Chesa Pilar , Adolf Günther R.
Published: February 2018

Focal adhesion kinase (FAK), a non-receptor tyrosine kinase, has attracted interest as a target for pharmacological intervention in malignant diseases. Here, we describe BI 853520, a novel ATP-competitive inhibitor distinguished by high potency and selectivity. In vitro, the compound inhibits FAK autophosphorylation in PC-3 prostate carcinoma cells with an IC50 of 1 nmol/L and blocks anchorage-independent proliferation of PC-3 cells with an EC50 of 3 nmol/L, whereas cells grown in conventional surface culture are 1000-fold less sensitive. In mice, the compound shows long half-life, high volume of distribution and high oral bioavailability; oral dosing of immunodeficient mice bearing subcutaneous PC-3 prostate adenocarcinoma xenografts resulted in rapid, long-lasting repression of FAK autophosphorylation in tumor tissue. Daily oral administration of BI 853520 to nude mice at doses of 50 mg/kg was well tolerated for prolonged periods of time. In a diverse panel of 16 subcutaneous adenocarcinoma xenograft models in nude mice, drug treatment resulted in a broad spectrum of outcomes, ranging from group median tumor growth inhibition values >100% and tumor regression in subsets of animals to complete lack of sensitivity. Biomarker analysis indicated that high sensitivity is linked to a mesenchymal tumor phenotype, initially defined by loss of E-cadherin expression and subsequently substantiated by gene set enrichment analysis. Further, we obtained microRNA expression profiles for 13 models and observed that hsa-miR-200c-3p expression is strongly correlated with efficacy (R2 = 0.889). BI 853520 is undergoing evaluation in early clinical trials.
PIM1 mediates epithelial-mesenchymal transition by targeting Smads and c-Myc in the nucleus and potentiates clear-cell renal-cell carcinoma oncogenesis
Authors: Zhao Bin , Liu Lei , Mao Jun , Zhang Zhiwei , Wang Qifei , Li Quanlin
Published: February 2018

Emerging evidence has shown that the PIM serine/threonine kinase family, including PIM1, PIM2 and PIM3, is associated with tumour progression towards metastasis. PIM1, an attractive molecular target, has been identified as a potential prognostic biomarker for haematological and epithelial malignancies. However, to date, the potential regulatory roles and molecular mechanisms by which PIM1 affects the development and progression of cancers, including clear-cell renal-cell carcinoma (ccRCC), remain largely unknown. Herein, we present the first evidence that PIM1 is aberrantly overexpressed in human ccRCC tissues and cell lines and positively correlated with human ccRCC progression. In our study, depletion of PIM1 attenuated ccRCC cell proliferation, colony formation, migration, invasion and angiogenesis, suggesting that PIM1 expression may be a cancer-promoting event in ccRCC. Mechanistically, we observed that PIM1 could interact with Smad2 or Smad3 in the nucleus and subsequently phosphorylate Smad2 and Smad3 to induce the expression of transcription factors, including ZEB1, ZEB2, Snail1, Snail2 and Twist, to promote epithelial-mesenchymal transition (EMT). In addition, PIM1-mediated phosphorylation of c-Myc activates the expression of the above transcription factors to synergistically promote EMT but does not activate Smads. Collectively, our results demonstrate that aberrant expression of PIM1 contributes to ccRCC development and progression. Moreover, our data reveal a potential molecular mechanism in which PIM1 mediates crosstalk between signalling pathways, including different Smad proteins and c-Myc, which target downstream transcription factors (ZEB1, ZEB2, Snail1, Snail2 and Twist) to trigger EMT. Together, our data suggest that PIM1 may be a potential therapeutic target for ccRCC patients.
Atm reactivation reverses ataxia telangiectasia phenotypes in vivo
Authors: Di Siena Sara , Campolo Federica , Gimmelli Roberto , Di Pietro Chiara , Marazziti Daniela , Dolci Susanna , Lenzi Andrea , Nussenzweig Andre , Pellegrini Manuela
Published: February 2018

Hereditary deficiencies in DNA damage signaling are invariably associated with cancer predisposition, immunodeficiency, radiation sensitivity, gonadal abnormalities, premature aging, and tissue degeneration. ATM kinase has been established as a central player in DNA double-strand break repair and its deficiency causes ataxia telangiectasia, a rare, multi-system disease with no cure. So ATM represents a highly attractive target for the development of novel types of gene therapy or transplantation strategies. Atm tamoxifen-inducible mouse models were generated to explore whether Atm reconstitution is able to restore Atm function in an Atm-deficient background. Body weight, immunodeficiency, spermatogenesis, and radioresistance were recovered in transgenic mice within 1 month from Atm induction. Notably, life span was doubled after Atm restoration, mice were protected from thymoma and no cerebellar defects were observed. Atm signaling was functional after DNA damage in vivo and in vitro. In summary, we propose a new Atm mouse model to investigate novel therapeutic strategies for ATM activation in ataxia telangiectasia disease.
High capacity in G protein-coupled receptor signaling
Authors: Keshelava Amiran , Solis Gonzalo P. , Hersch Micha , Koval Alexey , Kryuchkov Mikhail , Bergmann Sven , Katanaev Vladimir L.
Published: February 2018

G protein-coupled receptors (GPCRs) constitute a large family of receptors that activate intracellular signaling pathways upon detecting specific extracellular ligands. While many aspects of GPCR signaling have been uncovered through decades of studies, some fundamental properties, like its channel capacity—a measure of how much information a given transmission system can reliably transduce—are still debated. Previous studies concluded that GPCRs in individual cells could transmit around one bit of information about the concentration of the ligands, allowing only for a reliable on or off response. Using muscarinic receptor-induced calcium response measured in individual cells upon repeated stimulation, we show that GPCR signaling systems possess a significantly higher capacity. We estimate the channel capacity of this system to be above two, implying that at least four concentration levels of the agonist can be distinguished reliably. These findings shed light on the basic principles of GPCR signaling.
Spatial specificity of auxin responses coordinates wood formation
Authors: Brackmann Klaus , Qi Jiyan , Gebert Michael , Jouannet Virginie , Schlamp Theresa , Grünwald Karin , Wallner Eva-Sophie , Novikova Daria D. , Levitsky Victor G. , Agustí Javier , Sanchez Pablo , Lohmann Jan U. , Greb Thomas
Published: February 2018

Spatial organization of signalling events of the phytohormone auxin is fundamental for maintaining a dynamic transition from plant stem cells to differentiated descendants. The cambium, the stem cell niche mediating wood formation, fundamentally depends on auxin signalling but its exact role and spatial organization is obscure. Here we show that, while auxin signalling levels increase in differentiating cambium descendants, a moderate level of signalling in cambial stem cells is essential for cambium activity. We identify the auxin-dependent transcription factor ARF5/MONOPTEROS to cell-autonomously restrict the number of stem cells by directly attenuating the activity of the stem cell-promoting WOX4 gene. In contrast, ARF3 and ARF4 function as cambium activators in a redundant fashion from outside of WOX4-expressing cells. Our results reveal an influence of auxin signalling on distinct cambium features by specific signalling components and allow the conceptual integration of plant stem cell systems with distinct anatomies.
Two distinct domains contribute to the substrate acyl chain length selectivity of plant acyl-ACP thioesterase
Authors: Jing Fuyuan , Zhao Le , Yandeau-Nelson Marna D. , Nikolau Basil J.
Published: February 2018

The substrate specificity of acyl-ACP thioesterase (TE) plays an essential role in controlling the fatty acid profile produced by type II fatty acid synthases. Here we identify two groups of residues that synergistically determine different substrate specificities of two acyl-ACP TEs from Cuphea viscosissima (CvFatB1 and CvFatB2). One group (V194, V217, N223, R226, R227, and I268 in CvFatB2) is critical in determining the structure and depth of a hydrophobic cavity in the N-terminal hotdog domain that binds the substrate’s acyl moiety. The other group (255-RKLSKI-260 and 285-RKLPKL-289 in CvFatB2) defines positively charged surface patches that may facilitate binding of the ACP moiety. Mutagenesis of residues within these two groups results in distinct synthetic acyl-ACP TEs that efficiently hydrolyze substrates with even shorter chains (C4- to C8-ACPs). These insights into structural determinants of acyl-ACP TE substrate specificity are useful in modifying this enzyme for tailored fatty acid production in engineered organisms.
Rational design of a trispecific antibody targeting the HIV-1 Env with elevated anti-viral activity
Authors: Steinhardt James J. , Guenaga Javier , Turner Hannah L. , McKee Krisha , Louder Mark K. , O’Dell Sijy , Chiang Chi-I , Lei Lin , Galkin Andrey , Andrianov Alexander K. , A. Doria-Rose Nicole , Bailer Robert T. , Ward Andrew B. , Mascola John R. , Li Yuxing
Published: February 2018

HIV-1 broadly neutralizing antibodies (bNAbs) are being explored as passively administered therapeutic and preventative agents. However, the extensively diversified HIV-1 envelope glycoproteins (Env) rapidly acquire mutations to evade individual bNAbs in monotherapy regimens. The use of a “single” agent to simultaneously target distinct Env epitopes is desirable to overcome viral diversity. Here, we report the use of tandem single-chain variable fragment (ScFv) domains of two bNAbs, specific for the CD4-binding site and V3 glycan patch, to form anti-HIV-1 bispecific ScFvs (Bi-ScFvs). The optimal Bi-ScFv crosslinks adjacent protomers within one HIV-1 Env spike and has greater neutralization breadth than its parental bNAbs. Furthermore, the combination of this Bi-ScFv with a third bNAb recognizing the Env membrane proximal external region (MPER) results in a trispecific bNAb, which has nearly pan-isolate neutralization breadth and high potency. Thus, multispecific antibodies combining functional moieties of bNAbs could achieve outstanding neutralization capacity with augmented avidity.
‘Cyclical Bias’ in Microbiome Research Revealed by A Portable Germ-Free Housing System Using Nested Isolation
Authors: Rodriguez-Palacios Alexander , Aladyshkina Natalia , Ezeji Jessica C. , Erkkila Hailey L. , Conger Mathew , Ward John , Webster Joshua , Cominelli Fabio
Published: February 2018

Germ-Free (GF) research has required highly technical pressurized HEPA-ventilation anchored systems for decades. Herein, we validated a GF system that can be easily implemented and portable using Nested Isolation (NesTiso). GF-standards can be achieved housing mice in non-HEPA-static cages, which only need to be nested ‘one-cage-inside-another’ resembling ‘Russian dolls’. After 2 years of monitoring ~100,000 GF-mouse-days, NesTiso showed mice can be maintained GF for life (>1.3 years), with low animal daily-contamination-probability risk (1 every 867 days), allowing the expansion of GF research with unprecedented freedom and mobility. At the cage level, with 23,360 GF cage-days, the probability of having a cage contamination in NesTiso cages opened in biosafety hoods was statistically identical to that of opening cages inside (the ‘gold standard’) multi-cage pressurized GF isolators. When validating the benefits of using NesTiso in mouse microbiome research, our experiments unexpectedly revealed that the mouse fecal microbiota composition within the ‘bedding material’ of conventional SPF-cages suffers cyclical selection bias as moist/feces/diet/organic content (‘soiledness’) increases over time (e.g., favoring microbiome abundances ofBacillales, Burkholderiales, Pseudomonadales;and cultivableEnterococcus faecalisoverLactobacillus murinusandEscherichia coli), which in turn cyclically influences the gut microbiome dynamics of caged mice. Culture ‘co-streaking’ assays showed that cohoused mice exhibiting different fecal microbiota/hemolytic profiles in clean bedding (high-within-cage individual diversity) ‘cyclically and transiently appear identical’ (less diverse) as bedding soiledness increases, and recurs. Strategies are proposed to minimize this novel functional form of cyclical bedding-dependent microbiome selection bias.
Explaining combinatorial effects of mycotoxins Deoxynivalenol and Zearalenone in mice with urinary metabolomic profiling
Authors: Ji Jian , Zhu Pei , Blaženović Ivana , Cui Fangchao , Gholami Morteza , Sun Jiadi , Habimana Jean , Zhang Yinzhi , Sun Xiulan
Published: February 2018

Urine metabolic profiling of mice was conducted utilizing gas chromatography-mass spectrometry (GC-MS) to investigate the combinatory effect of mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) on the metabolism of the mice. Experiments were conducted by means of five-week-old mice which were individually exposed to 2 mg/kg DON, 20 mg/kg ZEN and the mixture of DON and ZEN (2 mg/kg and 20 mg/kg, respectively). The intragastric administration was applied for three weeks and urine samples were collected for metabolic analysis. Univariate and multivariate analysis were applied to data matrix processing along with respective pathway analysis by MetaMapp and CytoScape. The results showed that the combined DON and ZEN administration resulted in lower significant changes, compared to the individual mycotoxin treated groups verified by heatmap. Metabolic pathways network mapping indicated that the combined mycotoxins treated groups showed a little effect on the metabolites in most pathways, especially in glucose metabolism and its downstream amino acid metabolism. In glucose metabolism, the content of galactose, mannitol, galactonic acid, myo-inositol, tagatose was drastically down-regulated. Furthermore, the organic acids, pyruvate, and amino acids metabolism displayed the same phenomenon. In conclusion, the combined DON/ZEN administration might lead to an “antagonistic effect” in mice metabolism.
The Spanish version of Face-Name Associative Memory Exam (S-FNAME) performance is related to amyloid burden in Subjective Cognitive Decline
Authors: Sanabria Angela , Alegret Montserrat , Rodriguez-Gomez Octavio , Valero Sergi , Sotolongo-Grau Oscar , Monté-Rubio Gemma , Abdelnour Carla , Espinosa Ana , Ortega Gemma , Perez-Cordon Alba , Gailhajanet Anna , Hernandez Isabel , Rosende-Roca Maitee , Vargas Liliana , Mauleon Ana , Sanchez Domingo , Martin Elvira , Rentz Dorene M. , Lomeña Francisco , Ruiz Agustin , Tarraga Lluis , Boada Merce ,   The FACEHBI study group, Aguilera N. , Berthier M. , Buendia M. , Bullich S. , Campos F. , Cañabate P. , Cuevas C. , de Rojas I. , Diego S. , Gil S. , Giménez J. , Gismondi R. , Gómez-Chiari M. , Guitart M. , Hernández-Olasagarre B. , Ibarria M. , Lafuente A. , Martínez J. , Moreno M. , Moreno-Grau S. , Núñez L. , Orellana A. , Páez A. , Pancho A. , Pavía J. , Pelejà E. , Pérez-Grijalba V. , Pesini P. , Preckler S. , Romero J. , Ruiz S. , Santos M. , Sarasa M. , Tejero M. A. , Torres M. , Vivas A.
Published: February 2018

The Face-Name Associative Memory Exam (FNAME) is a paired associative memory test created to detect memory deficits in individuals with preclinical Alzheimer’s disease (AD). Worse performance on FNAME in cognitively healthy individuals were found related to higher amyloid beta (Aβ) burden measured with Positron-Emission-Tomography using 11C-PiB (PiB-PET). We previously reported normative data of a Spanish version of FNAME (S-FNAME) in cognitively healthy Spanish-speaking subjects. The aim of the present study was to determine whether performance on S-FNAME was associated with Aβ burden in subjective cognitive decline (SCD) individuals. 200 SCD subjects received neurological and neuropsychological assessments, including the S-FNAME and the Word List task from the Wechsler-Memory-Scale-III (WMS-III). Moreover, they received an MRI and (18)F-Florbetaben Positron-Emission-Tomography (FBB-PET) to measure Aβ burden. Three cognitive factor composites were derived for the episodic memory measures (face-name [SFN-N], face-occupation [SFN-O] and WMS-III) to determine whether episodic memory performance was related to Aβ deposition. Higher global Aβ deposition was significantly related to worse performance on SFN-N but not with SFN-O or WMS-III Composite. Moreover, worse SFN-N performance was significantly related to higher Aβ deposition in bilateral  Posterior Cingulate Cortex. The S-FNAME may be a promising neuropsychological tool for detecting SCD individuals with preclinical AD.
Towards in cellulo virus crystallography
Authors: Duyvesteyn Helen M. E. , Ginn Helen M. , Pietilä Maija K. , Wagner Armin , Hattne Johan , Grimes Jonathan M. , Hirvonen Elina , Evans Gwyndaf , Parsy Marie-Laure , Sauter Nicholas K. , Brewster Aaron S. , Huiskonen Juha T. , Stuart David I. , Sutton Geoff , Bamford Dennis H.
Published: February 2018

Viruses are a significant threat to both human health and the economy, and there is an urgent need for novel anti-viral drugs and vaccines. High-resolution viral structures inform our understanding of the virosphere, and inspire novel therapies. Here we present a method of obtaining such structural information that avoids potentially disruptive handling, by collecting diffraction data from intact infected cells. We identify a suitable combination of cell type and virus to accumulate particles in the cells, establish a suitable time point where most cells contain virus condensates and use electron microscopy to demonstrate that these are ordered crystalline arrays of empty capsids. We then use an X-ray free electron laser to provide extremely bright illumination of sub-micron intracellular condensates of bacteriophage phiX174 inside living Escherichia coli at room temperature. We have been able to collect low resolution diffraction data. Despite the limited resolution and completeness of these initial data, due to a far from optimal experimental setup, we have used novel methodology to determine a putative space group, unit cell dimensions, particle packing and likely maturation state of the particles.
Cognitive control over memory – individual differences in memory performance for emotional and neutral material
Authors: Wierzba M. , Riegel M. , Wypych M. , Jednoróg K. , Grabowska A. , Marchewka A.
Published: February 2018

It is widely accepted that people differ in memory performance. The ability to control one’s memory depends on multiple factors, including the emotional properties of the memorized material. While it was widely demonstrated that emotion can facilitate memory, it is unclear how emotion modifies our ability to suppress memory. One of the reasons for the lack of consensus among researchers is that individual differences in memory performance were largely neglected in previous studies. We used the directed forgetting paradigm in an fMRI study, in which subjects viewed neutral and emotional words, which they were instructed to remember or to forget. Subsequently, subjects’ memory of these words was tested. Finally, they assessed the words on scales of valence, arousal, sadness and fear. We found that memory performance depended on instruction as reflected in the engagement of the lateral prefrontal cortex (lateral PFC), irrespective of emotional properties of words. While the lateral PFC engagement did not differ between neutral and emotional conditions, it correlated with behavioural performance when emotional – as opposed to neutral – words were presented. A deeper understanding of the underlying brain mechanisms is likely to require a study of individual differences in cognitive abilities to suppress memory.
JMJD-1.2 controls multiple histone post-translational modifications in germ cells and protects the genome from replication stress
Authors: Myers Toshia R. , Amendola Pier Giorgio , Lussi Yvonne C. , Salcini Anna Elisabetta
Published: February 2018

Post-translational modifications of histones, constitutive components of chromatin, regulate chromatin compaction and control all DNA-based cellular processes. C. elegans JMJD-1.2, a member of the KDM7 family, is a demethylase active towards several lysine residues on Histone 3 (H3), but its contribution in regulating histone methylation in germ cells has not been fully investigated. Here, we show that jmjd-1.2 is expressed abundantly in the germline where it controls the level of histone 3 lysine 9, lysine 23 and lysine 27 di-methylation (H3K9/K23/K27me2) both in mitotic and meiotic cells. Loss of jmjd-1.2 is not associated with major defects in the germ cells in animals grown under normal conditions or after DNA damage induced by UV or ionizing irradiation. However, jmjd-1.2 mutants are more sensitive to replication stress and the progeny of mutant animals exposed to hydroxyurea show increased embryonic lethality and mutational rate, compared to wild-type. Thus, our results suggest a role for jmjd-1.2 in the maintenance of genome integrity after replication stress and emphasize the relevance of the regulation of histone methylation in genomic stability.
ASC- and caspase-8-dependent apoptotic pathway diverges from the NLRC4 inflammasome in macrophages
Authors: Lee Bettina L. , Mirrashidi Kathleen M. , Stowe Irma B. , Kummerfeld Sarah K. , Watanabe Colin , Haley Benjamin , Cuellar Trinna L. , Reichelt Michael , Kayagaki Nobuhiko
Published: February 2018

The NLRC4 inflammasome recognizes bacterial flagellin and components of the type III secretion apparatus. NLRC4 stimulation leads to caspase-1 activation followed by a rapid lytic cell death known as pyroptosis. NLRC4 is linked to pathogen-free auto-inflammatory diseases, suggesting a role for NLRC4 in sterile inflammation. Here, we show that NLRC4 activates an alternative cell death program morphologically similar to apoptosis in caspase-1-deficient BMDMs. By performing an unbiased genome-wide CRISPR/Cas9 screen with subsequent validation studies in gene-targeted mice, we highlight a critical role for caspase-8 and ASC adaptor in an alternative apoptotic pathway downstream of NLRC4. Furthermore, caspase-1 catalytically dead knock-in (Casp1 C284A KI) BMDMs genetically segregate pyroptosis and apoptosis, and confirm that caspase-1 does not functionally compete with ASC for NLRC4 interactions. We show that NLRC4/caspase-8-mediated apoptotic cells eventually undergo plasma cell membrane damage in vitro, suggesting that this pathway can lead to secondary necrosis. Unexpectedly, we found that DFNA5/GSDME, a member of the pore-forming gasdermin family, is dispensable for the secondary necrosis that follows NLRC4-mediated apoptosis in macrophages. Together, our data confirm the existence of an alternative caspase-8 activation pathway diverging from the NLRC4 inflammasome in primary macrophages.