bluuebunny/crossref_metadata_2025 huggingface dataset:
Striking the Balance between Human Rights Online and State Security Concerns: The Russian Way in a Comparative Context
Authors: Pollicino Oreste , Soldatov Oleg
Published: March 2019

In pursuit of solutions to curb cybercrime, legislators engage in an analysis proportionally weighing freedom of expression and other societal interests. The balance between the two concepts differs dramatically across different jurisdictions. This Article looks into a widely discussed legislative package regulating the online domain, enacted by the Sixth Convocation of the Russian Parliament (2011–2016)—the State Duma. The authors operate under the assumption that the Russian approach might have a broad spillover effect. With this in mind, the authors outline the current status quo regarding Internet regulations in the EU, disentangle and contextualize the legislation under scrutiny, emphasize Russian influence over Eastern European countries, and describe the tumultuous relationship between the Russian Federation and the European Court of Human Rights.
Identifying German Legal Approaches to Terror—How the Constitution Shapes Legislation Allowing the Shooting Down of a Hijacked Plane
Authors: Müller Lukas Frederik
Published: March 2019

In the face of a recent increase in the number of Islamic terrorist attacks all over the globe—particularly in Europe—counterterrorism has become a main field of political and legal activity once again. In this turbulent context, this Article will look back on how German legislation has reacted to the first significant occurrences of Islamic terrorism in Europe and the United States in the 2000s and how the Constitution has been put in place in the conflict between individual and collective interests by the Federal Constitutional Court in this context. From the wide range of counterterrorism measures that were significantly shaped along the principles of the Constitution since then, this Article takes a detailed look at the legal issues relating to a very specific measure—legislation allowing the shooting down of a hijacked plane. This issue has been vigorously debated among politicians, scholars, and courts in Germany since 2004. It constitutes a vital element of the fundamental legal structures that governs state action that violates certain legal interests in order to protect conflicting ones—a key question for other recent strategies of terrorism as well.
Developments in AKP Policy Toward Religion and Homogeneity
Authors: Watters Samuel W.
Published: March 2019

Critics of the Turkish interpretation of secularism,laiklik, describe it as authoritarian and repressive. Indeed, rather than establish state neutrality toward religion,laiklikhistorically entailed state control of Islam, the religion of the vast majority of the Turkish population, and the exclusion of religion from the public sphere in an effort to control religious belief and identity. Many, including leaders in the ruling AKP, assert, though, that recent reforms herald a move away from this model of control toward a secularism defined by state neutrality toward religion. To determine whether this transformation is actually occurring, I evaluate, based on Turkish language sources, the recent reforms under the AKP using the framework of the secularized state described by the German legal scholar Ernst-Wolfgang Böckenförde. Because of its significant role in implementing Turkish policies toward religion, I evaluate these reforms by analyzing developments in the programming and messaging of the Directorate of Religious Affairs (Diyanet) under the AKP. I find little evidence thatlaiklikis transitioning to a state neutrality toward religion. Rather, the AKP has coupled a greater presence of religion in the public sphere with expanding state authority in religious programming and messaging. Although these reforms reflect a transformation in Turkish nation-building policies, they maintain the state control of religion that separateslaiklikfrom neutral secularism.
An Interdisciplinary Approach for Comparative Lawyers: Insights from the Fast-Moving Field of Law and Technology
Authors: Guerra Giorgia
Published: March 2019

Focused on comparative law and technology, the Article demonstrates the importance of recent advances of comparative law on interdisciplinary research and argues that comparative law becomes essential to broaden this approach, when facing modern techno-scientific issues. Technological convergence has consequences not only for human rights and the way they can be exercised, but also for the fundamental concept of what characterizes a human being. Social perceptions of risks and geopolitical contexts deeply affect the legal approach to uncertainty. Converging technologies thus challenge us to develop a new, wider perspective on the concept of safety. Comparative law research is functional to deal with technological issues characterized by a cross-national and cross border nature. Additionally, it helps to integrate non-legal knowledge into legal culture. It thus becomes an essential pre-condition to answer complex and innovative questions. Even when research questions focus on the sole legal consequences of technology innovation, this cannot be done without a broader vision of the multidisciplinary problem technology offers. In fact, such a problem appears like an elephant: a unique perspective will not faithfully represent the real image. The Article focuses on a case-study: robotics. An emerging techno-scientific arena where neuro-sciences, informatics, and other disciplines, however, will deeply orientate the analysis of all the legal issues.
The Many Uses of Anti-Money Laundering Regulation—Over Time and into the Future
Authors: Bergström Maria
Published: March 2019

Given the fast development of the field of AML Regulation, this Article aims to answer the following questions: First, how is money laundering dealt with and regulated on the EU level? Second, to which legal concerns do the chosen regulatory strategy give rise? Accordingly, this Article provides an overview of the various regulatory strategies in the global and EU regional AML Regime while at the same time points out some of the most pressing legal concerns in AML Regulation. These include the blurred line between administrative and criminal law measures and the protection of individual rights and fundamental freedoms including data protection and privacy issues in administrative and criminal law contexts respectively. Although briefly mentioning the global and international context, the focus of this Article is the EU regulatory action, its outcome and critique, and possible future.
Some Reflections on Dignity as an Alternative Legal Concept in Data Protection Regulation
Authors: de Hingh Anne
Published: March 2019

As the use of the Internet and online platforms grows, the scale of collecting and processing personal data and turnovers have increased correspondingly.1At the same time, public awareness about the Internet turning into a genuine profiling and advertisement machine, as well as a powerful surveillance instrument, grows. More people today are concerned about the ways in which public and private actors store and use private information. Many individuals note that they lose sight of the consequences once they give consent to the collection of their sometimes most intimate personal data. The Snowden revelations and the recent Facebook and Cambridge Analytica scandal have only reinforced this public awareness.

Objections against these data processing practices cannot be explained as breaches of data protection or privacy regulation alone. In this Article, it is argued that recently passed regulations fail to solve the unease of data subjects as other, more fundamental values are at stake here. A different or complementary ethical and legal framework is needed to interpret this generally felt unease vis-à-vis current data practices and secondly to confront future developments on the data market. The concept of human dignity may be a helpful perspective in this respect. In the context of data processing, human dignity is generally interpreted in a quite specific manner, such as contributing to the empowerment and self-determination of autonomous individuals. It can be argued, however, that human dignity—in the context of the commodification and commoditization of online personal data—should be seen in a different, quite opposite, light. In sum, future regulation of privacy and data protection attention should shift towards more constraining dimensions of human dignity.
The Carles Puigdemont Case: Europe's Criminal Law in the Crisis of Confidence
Authors: Braum Stefan
Published: March 2019

The case of Carles Puigdemont underlines that European criminal law is in a crisis of confidence. The Higher Regional Court of Schleswig-Holstein has rejected a Spanish European arrest warrant for the criminal offence of rebellion because it lacks double criminality. It applied German law de lege artis without, however, questioning the European legal framework. The case would have provided an opportunity to refer the matter to the European Court of Justice for a preliminary ruling in order to further specify the European law criteria of double criminality. This would have been the adequate legal response to a politically explosive case. In the end, the Spanish judiciary sees itself disavowed and the system of the European arrest warrant called into question.
Determining the “Private Law” Nature: A Comparison Between Customary State Practice and Turkish Court Rulings
Authors: Gökçe Yasir
Published: March 2019

This Article uses a combination of doctrinal and comparative analyses to elucidate the most widely-used criteria according to which courts characterize an act as having a private-law nature. Further, this Article identifies the extent to which the Turkish State practice has embraced those criteria. To that end, this Article extracts two such widely-used criteria from patterns in customary State practice. In the end, the Article points out the extent to which those criteria are prevalent throughout the Turkish court rulings and concludes that Turkish caselaw is generally in alignment with the customary State practice in terms of the criteria used to determine a State act's private-law nature.
Alexander Somek'sThe Cosmopolitan Constitution
Authors: Tschorne Samuel I.
Published: March 2019

The Cosmopolitan Constitutionis an intriguing and puzzling book. In particular, the book has the uncanny ability to render fresh what is for the constitutional theorist familiar territory such as the debate on judicial supremacy and the counter-majoritarian difficulty, the expansion of the proportionality principle, etc. In fact, one of the most interesting aspects of its overarching argument is that given our present conditions—such as those of the cosmopolitan constitution or constitutionalism 3.0—we should be increasingly plagued by self-doubt, at least to the extent that we are to remain committed to the tenets that once animated our constitutional traditions. This implies that the partial sense of disorientation with which one is left after reading the book is deliberately provoked precisely because of the main insights it has to offer.
Re-Imagining the Cosmopolitan Constitution: A Comment on Alexander Somek'sThe Cosmopolitan Constitution
Authors: Dani Marco
Published: March 2019

This comment engages with Somek's account for the cosmopolitan constitution for two distinct purposes. It first builds on its inherent ambivalence to argue that hitherto Europe has experienced at least two different versions of the cosmopolitan constitution. Whereas in the formative period of the process of European integration, the cosmopolitan constitution manifested itself prevailingly in its more benign and democracy-enhancing political face, since the end of the 1970s it has developed a more controversial and democracy-inhibiting administrative profile. Secondly, the comment rejects as potentially regressive the proposal of redressing the biases inherent in the contemporary legal and political order by reviving the idea of a Machiavellian mixed constitution. To fulfill the promise of emancipation inspiring constitutionalism, the cosmopolitan constitution cannot segregate in separate institutions ordinary citizens and a market elite. Rather, the cosmopolitan constitution should be re-imagined with a view to reassert its original democracy-enhancing spirit and adapt it to the evolving circumstances of economic and political interdependence.
Geologic Analyses of the Causes of Morphological Variations in Lunar Craters Within the Simple‐to‐Complex Transition
Authors: Chandnani M. , Herrick R. R. , Kramer G. Y.
Published: March 2019

The diameter range of 15 to 20 km is within the transition from simple to complex impact craters located on the Moon. This range spans roughly a factor of 3 in impact energy for the same impactor speed, composition, and trajectory angle. We analyzed the global population of well‐preserved craters in this size range in order to assess the effects of target and impactor properties on crater shapes and morphologies. We observed that within this narrow diameter range, simple craters are confined to the highlands, and complex craters are more abundant in the mare. We found unusually deep craters around the highlands‐mare boundaries and favor the hypothesis that they form by impact cratering on high‐porosity terrain. We infer that target properties primarily contribute to the observed morphological variations in the craters. Simple crater formation is favored by a terrain that is more homogeneous in strength and topography, while transitional and complex crater formation is aided by heterogeneity in lithology, topography, or strength, or a combination of these parameters. Clearly visible impact melt deposits in a small percentage of simple craters, and two cases of craters differing in morphologies from their nearest neighbors in similar geologic settings, suggest that variation in impactor properties such as impact velocity and impactor size may have some role in causing morphological differences between similar‐sized craters.
Verification and rectification of cell type-specific splicing of a Seckel syndrome-associated ATR mutation using iPS cell model
Authors: Ichisima Jose , Suzuki Naoya M. , Samata Bumpei , Awaya Tomonari , Takahashi Jun , Hagiwara Masatoshi , Nakahata Tatsutoshi , Saito Megumu K.
Published: March 2019

Seckel syndrome (SS) is a rare spectrum of congenital severe microcephaly and dwarfism. One SS-causative gene is Ataxia Telangiectasia and Rad3-Related Protein (ATR), and ATR (c.2101 A>G) mutation causes skipping of exon 9, resulting in a hypomorphic ATR defect. This mutation is considered the cause of an impaired response to DNA replication stress, the main function of ATR, contributing to the pathogenesis of microcephaly. However, the precise behavior and impact of this splicing defect in human neural progenitor cells (NPCs) is unclear. To address this, we established induced pluripotent stem cells (iPSCs) from fibroblasts carrying the ATR mutation and an isogenic ATR-corrected counterpart iPSC clone. SS-patient-derived iPSCs (SS-iPSCs) exhibited cell type-specific splicing; exon 9 was dominantly skipped in fibroblasts and iPSC-derived NPCs, but it was included in undifferentiated iPSCs and definitive endodermal cells. SS-iPSC-derived NPCs (SS-NPCs) showed distinct expression profiles from ATR non-mutated NPCs with negative enrichment of neuronal genesis-related gene sets. In SS-NPCs, abnormal mitotic spindles occurred more frequently than in gene-corrected counterparts, and the alignment of NPCs in the surface of the neurospheres was perturbed. Finally, we tested several splicing-modifying compounds and found that TG003, a CLK1 inhibitor, could pharmacologically rescue the exon 9 skipping in SS-NPCs. Treatment with TG003 restored the ATR kinase activity in SS-NPCs and decreased the frequency of abnormal mitotic events. In conclusion, our iPSC model revealed a novel effect of the ATR mutation in mitotic processes of NPCs and NPC-specific missplicing, accompanied by the recovery of neuronal defects using a splicing rectifier.
Integrating microsystems with metamaterials towards metadevices
Authors: Zhao Xiaoguang , Duan Guangwu , Li Aobo , Chen Chunxu , Zhang Xin
Published: March 2019

Electromagnetic metamaterials, which are a major type of artificially engineered materials, have boosted the development of optical and photonic devices due to their unprecedented and controllable effective properties, including electric permittivity and magnetic permeability. Metamaterials consist of arrays of subwavelength unit cells, which are also known as meta-atoms. Importantly, the effective properties of metamaterials are mainly determined by the geometry of the constituting subwavelength unit cells rather than their chemical composition, enabling versatile designs of their electromagnetic properties. Recent research has mainly focused on reconfigurable, tunable, and nonlinear metamaterials towards the development of metamaterial devices, namely, metadevices, via integrating actuation mechanisms and quantum materials with meta-atoms. Microelectromechanical systems (MEMS), or microsystems, provide powerful platforms for the manipulation of the effective properties of metamaterials and the integration of abundant functions with metamaterials. In this review, we will introduce the fundamentals of metamaterials, approaches to integrate MEMS with metamaterials, functional metadevices from the synergy, and outlooks for metamaterial-enabled photonic devices.
Human colon cancer cells highly express myoferlin to maintain a fit mitochondrial network and escape p53-driven apoptosis
Authors: Rademaker Gilles , Costanza Brunella , Bellier Justine , Herfs Michael , Peiffer Raphaël , Agirman Ferman , Maloujahmoum Naïma , Habraken Yvette , Delvenne Philippe , Bellahcène Akeila , Castronovo Vincent , Peulen Olivier
Published: March 2019

Colon adenocarcinoma is the third most commonly diagnosed cancer and the second deadliest one. Metabolic reprogramming, described as an emerging hallmark of malignant cells, includes the predominant use of glycolysis to produce energy. Recent studies demonstrated that mitochondrial electron transport chain inhibitor reduced colon cancer tumour growth. Accumulating evidence show that myoferlin, a member of the ferlin family, is highly expressed in several cancer types, where it acts as a tumour promoter and participates in the metabolic rewiring towards oxidative metabolism. In this study, we showed that myoferlin expression in colon cancer lesions is associated with low patient survival and is higher than in non-tumoural adjacent tissue. Human colon cancer cells silenced for myoferlin exhibit a reduced oxidative phosphorylation activity associated with mitochondrial fission leading, ROS accumulation, decreased cell growth, and increased apoptosis. We observed the triggering of a DNA damage response culminating to a cell cycle arrest in wild-type p53 cells. The use of a p53 null cell line or a compound able to restore p53 activity (Prima-1) reverted the effects induced by myoferlin silencing, confirming the involvement of p53. The recent identification of a compound interacting with a myoferlin C2 domain and bearing anticancer potency identifies, together with our demonstration, this protein as a suitable new therapeutic target in colon cancer.
Forming global estimates of self-performance from local confidence
Authors: Rouault Marion , Dayan Peter , Fleming Stephen M.
Published: March 2019

Metacognition, the ability to internally evaluate our own cognitive performance, is particularly useful since many real-life decisions lack immediate feedback. While most previous studies have focused on the construction of confidence at the level of single decisions, little is known about the formation of “global” self-performance estimates (SPEs) aggregated from multiple decisions. Here, we compare the formation of SPEs in the presence and absence of feedback, testing a hypothesis that local decision confidence supports the formation of SPEs when feedback is unavailable. We reveal that humans pervasively underestimate their performance in the absence of feedback, compared to a condition with full feedback, despite objective performance being unaffected. We find that fluctuations in confidence contribute to global SPEs over and above objective accuracy and reaction times. Our findings create a bridge between a computation of local confidence and global SPEs, and support a functional role for confidence in higher-order behavioral control.
Genome-wide association study in frontal fibrosing alopecia identifies four susceptibility loci including HLA-B*07:02
Authors: Tziotzios Christos , Petridis Christos , Dand Nick , Ainali Chrysanthi , Saklatvala Jake R. , Pullabhatla Venu , Onoufriadis Alexandros , Pramanik Rashida , Baudry David , Lee Sang Hyuck , Wood Kristie , Liu Lu , Seegobin Seth , Michelotti Gregory A. , Lwin Su M. , Christou Evangelos A. A. , Curtis Charles J. , de Rinaldis Emanuele , Saxena Alka , Holmes Susan , Harries Matthew , Palamaras Ioulios , Cunningham Fiona , Parkins Gregory , Kaur Manjit , Farrant Paul , McDonagh Andrew , Messenger Andrew , Jones Jennifer , Jolliffe Victoria , Ali Iaisha , Ardern-Jones Michael , Mitchell Charles , Burrows Nigel , Atkar Ravinder , Banfield Cedric , Alexandroff Anton , Champagne Caroline , Cooper Hywel L. , Vañó-Galván Sergio , Molina-Ruiz Ana Maria , Perez Nerea Ormaechea , Patel Girish K. , Macbeth Abby , Page Melanie , Bryden Alyson , Mowbray Megan , Wahie Shyamal , Armstrong Keith , Cooke Nicola , Goodfield Mark , Man Irene , de Berker David , Dunnill Giles , Takwale Anita , Rao Archana , Siah Tee-Wei , Sinclair Rodney , Wade Martin S. , Dlova Ncoza C. , Setterfield Jane , Lewis Fiona , Bhargava Kapil , Kirkpatrick Niall , Estivill Xavier , Stefanato Catherine M. , Flohr Carsten , Spector Timothy , Watt Fiona M. , Smith Catherine H. , Barker Jonathan N. , Fenton David A. , Simpson Michael A. , McGrath John A.
Published: March 2019

Frontal fibrosing alopecia (FFA) is a recently described inflammatory and scarring type of hair loss affecting almost exclusively women. Despite a dramatic recent increase in incidence the aetiopathogenesis of FFA remains unknown. We undertake genome-wide association studies in females from a UK cohort, comprising 844 cases and 3,760 controls, a Spanish cohort of 172 cases and 385 controls, and perform statistical meta-analysis. We observe genome-wide significant association with FFA at four genomic loci: 2p22.2, 6p21.1, 8q24.22 and 15q2.1. Within the 6p21.1 locus, fine-mapping indicates that the association is driven by theHLA-B*07:02 allele. At 2p22.1, we implicate a putative causal missense variant inCYP1B1, encoding the homonymous xenobiotic- and hormone-processing enzyme. Transcriptomic analysis of affected scalp tissue highlights overrepresentation of transcripts encoding components of innate and adaptive immune response pathways. These findings provide insight into disease pathogenesis and characterise FFA as a genetically predisposed immuno-inflammatory disorder driven byHLA-B*07:02.
Analyzing machine learning models to accelerate generation of fundamental materials insights
Authors: Umehara Mitsutaro , Stein Helge S. , Guevarra Dan , Newhouse Paul F. , Boyd David A. , Gregoire John M.
Published: March 2019

Machine learning for materials science envisions the acceleration of basic science research through automated identification of key data relationships to augment human interpretation and gain scientific understanding. A primary role of scientists is extraction of fundamental knowledge from data, and we demonstrate that this extraction can be accelerated using neural networks via analysis of the trained data model itself rather than its application as a prediction tool. Convolutional neural networks excel at modeling complex data relationships in multi-dimensional parameter spaces, such as that mapped by a combinatorial materials science experiment. Measuring a performance metric in a given materials space provides direct information about (locally) optimal materials but not the underlying materials science that gives rise to the variation in performance. By building a model that predicts performance (in this case photoelectrochemical power generation of a solar fuels photoanode) from materials parameters (in this case composition and Raman signal), subsequent analysis of gradients in the trained model reveals key data relationships that are not readily identified by human inspection or traditional statistical analyses. Human interpretation of these key relationships produces the desired fundamental understanding, demonstrating a framework in which machine learning accelerates data interpretation by leveraging the expertize of the human scientist. We also demonstrate the use of neural network gradient analysis to automate prediction of the directions in parameter space, such as the addition of specific alloying elements, that may increase performance by moving beyond the confines of existing data.
Automated measurements of greenhouse gases fluxes from tree stems and soils: magnitudes, patterns and drivers
Authors: Barba Josep , Poyatos Rafael , Vargas Rodrigo
Published: March 2019

Tree stems exchange CO2, CH4 and N2O with the atmosphere but the magnitudes, patterns and drivers of these greenhouse gas (GHG) fluxes remain poorly understood. Our understanding mainly comes from static-manual measurements, which provide limited information on the temporal variability and magnitude of these fluxes. We measured hourly CO2, CH4 and N2O fluxes at two stem heights and adjacent soils within an upland temperate forest. We analyzed diurnal and seasonal variability of fluxes and biophysical drivers (i.e., temperature, soil moisture, sap flux). Tree stems were a net source of CO2 (3.80 ± 0.18 µmol m−2 s−1; mean ± 95% CI) and CH4 (0.37 ± 0.18 nmol m−2 s−1), but a sink for N2O (−0.016 ± 0.008 nmol m−2 s−1). Time series analysis showed diurnal temporal correlations between these gases with temperature or sap flux for certain days. CO2 and CH4 showed a clear seasonal pattern explained by temperature, soil water content and sap flux. Relationships between stem, soil fluxes and their drivers suggest that CH4 for stem emissions could be partially produced belowground. High-frequency measurements demonstrate that: a) tree stems exchange GHGs with the atmosphere at multiple time scales; and b) are needed to better estimate fluxes magnitudes and understand underlying mechanisms of GHG stem emissions.
Synthesis of the extended phenacene molecules, [10]phenacene and [11]phenacene, and their performance in a field-effect transistor
Authors: Okamoto Hideki , Hamao Shino , Eguchi Ritsuko , Goto Hidenori , Takabayashi Yasuhiro , Yen Paul Yu-Hsiang , Liang Luo Uei , Chou Chia-Wei , Hoffmann Germar , Gohda Shin , Sugino Hisako , Liao Yen-Fa , Ishii Hirofumi , Kubozono Yoshihiro
Published: March 2019

The [10]phenacene and [11]phenacene molecules have been synthesized using a simple repetition of Wittig reactions followed by photocyclization. Sufficient amounts of [10]phenacene and [11]phenacene were obtained, and thin-film FETs using these molecules have been fabricated with SiO2 and ionic liquid gate dielectrics. These FETs operated in p-channel. The averaged measurements of field-effect mobility, <μ>, were 3.1(7) × 10−2 and 1.11(4) × 10−1 cm2 V−1 s−1, respectively, for [10]phenacene and [11]phenacene thin-film FETs with SiO2 gate dielectrics. Furthermore, [10]phenacene and [11]phenacene thin-film electric-double-layer (EDL) FETs with ionic liquid showed low-voltage p-channel FET properties, with <μ> values of 3(1) and 1(1) cm2 V−1 s−1, respectively. This study also discusses the future utility of the extremely extended π-network molecules [10]phenacene and [11]phenacene as the active layer of FET devices, based on the experimental results obtained.
Cytotoxicity profiling of deep eutectic solvents to human skin cells
Authors: Macário I. P. E. , Oliveira H. , Menezes A. C. , Ventura S. P. M. , Pereira J. L. , Gonçalves A. M. M. , Coutinho J. A. P. , Gonçalves F. J. M.
Published: March 2019

The tailor-made character of deep eutectic solvents (DES) turns them very attractive to be used in several applications, including in health-related areas such as pharmaceutical, nutraceutical, and cosmetic industries. However, although DES has been touted as “green” solvents, several works proved that their potential toxicity should not be neglected. Using the premise of DES applicability in the cosmetic and pharmaceutical sectors, we chose two cell lines to work as a skin model (keratinocytes HaCaT and tumor melanocytes MNT-1), to assess DES cytotoxicity. The effect of three different hydrogen bond acceptors (HBA) ([Chol]Cl, [N1111]Cl and [N4444]Cl) and three different hydrogen bond donors (HBD) (hexanoic and butanoic acid, ethylene glycol, 1-propanol and urea) were evaluated through a common viability assay (MTT assay). Results were promising since [Chol]Cl and [N1111]Cl- based DES showed good biocompatibility for the tested cells. [N4444]Cl-based DES, however, showed cytotoxicity for both cell lines, with the HBA being the driver of the toxicity. Interestingly, some compounds increased cell viability in the HaCaT cell line, namely [Chol]Cl, ethylene glycol, hexanoic acid, urea, and all [Chol]Cl and [N1111]Cl-based DES and should be considered as targets for future studies. These results highlight their possible use in cosmetic or pharmaceutical formulations.
RNA Stabilizes Transcription-Dependent Chromatin Loops Induced By Nuclear Hormones
Authors: Pezone Antonio , Zuchegna Candida , Tramontano Alfonso , Romano Antonella , Russo Giusi , de Rosa Mariarosaria , Vinciguerra Maria , Porcellini Antonio , Gottesman Max E. , Avvedimento Enrico Vittorio
Published: March 2019

We show that transcription induced by nuclear receptors for estrogen (E2) or retinoic acid (RA) is associated with formation of chromatin loops that juxtapose the 5’ end (containing the promoter) with the enhancer and the 3′ polyA addition site of the target gene. We find three loop configurations which change as a function of time after induction: 1. RA or E2-induced loops which connect the 5′ end, the enhancer and the 3′ end of the gene, and are stabilized by RNA early after induction; 2. E2-independent loops whose stability does not require RNA; 3. Loops detected only by treatment of chromatin with RNAse H1 prior to hormonal induction. RNAse H1 digests RNA that occludes the relevant restriction sites, thus preventing detection of these loops. R-loops at the 5′ and 3′ ends of the RA or E2-target genes were demonstrated by immunoprecipitation with anti-DNA-RNA hybrid antibodies as well as by sensitivity to RNAse H1. The cohesin RAD21 subunit is preferentially recruited to the target sites upon RA or E2 induction of transcription. RAD21 binding to chromatin is eliminated by RNAse H1. We identified E2-induced and RNase H1-sensitive antisense RNAs located at the 5′ and 3′ ends of the E2-induced transcription unit which stabilize the loops and RAD21 binding to chromatin. This is the first report of chromatin loops that form after gene induction that are maintained by RNA:DNA hybrids.
Magnetically guided non-invasive CRISPR-Cas9/gRNA delivery across blood-brain barrier to eradicate latent HIV-1 infection
Authors: Kaushik Ajeet , Yndart Adriana , Atluri Venkata , Tiwari Sneham , Tomitaka Asahi , Gupta Purnima , Jayant Rahul Dev , Alvarez-Carbonell David , Khalili Kamel , Nair Madhavan
Published: March 2019

CRISPR-Cas9/gRNA exhibits therapeutic efficacy against latent human immunodeficiency virus (HIV) genome but the delivery of this therapeutic cargo to the brain remains as a challenge. In this research, for the first time, we demonstrated magnetically guided non-invasive delivery of a nano-formulation (NF), composed of Cas9/gRNA bound with magneto-electric nanoparticles (MENPs), across the blood-brain barrier (BBB) to inhibit latent HIV-1 infection in microglial (hμglia)/HIV (HC69) cells. An optimized ac-magnetic field of 60 Oe was applied on NF to release Cas9/gRNA from MENPs surface and to facilitate NF cell uptake resulting in intracellular release and inhibition of HIV. The outcomes suggested that developed NF reduced HIV-LTR expression significantly in comparison to unbound Cas9/gRNA in HIV latent hμglia/HIV (HC69) cells. These findings were also validated qualitatively using fluorescence microscopy to assess NF efficacy against latent HIV in the microglia cells. We believe that CNS delivery of NF (CRISPR/Cas9-gRNA-MENPs) across the BBB certainly will have clinical utility as future personalized nanomedicine to manage neuroHIV/AIDS.
Global response of diacylglycerol kinase towards substrate binding observed by 2D and 3D MAS NMR
Authors: Möbius Kristin , Kazemi Sina , Güntert Peter , Jakob Andreas , Heckel Alexander , Becker-Baldus Johanna , Glaubitz Clemens
Published: March 2019

Escherichia coli diacylglycerol kinase (DGK) is an integral membrane protein, which catalyses the ATP-dependent phosphorylation of diacylglycerol (DAG) to phosphatic acid (PA). It is a unique trimeric enzyme, which does not share sequence homology with typical kinases. It exhibits a notable complexity in structure and function despite of its small size. Here, chemical shift assignment of wild-type DGK within lipid bilayers was carried out based on 3D MAS NMR, utilizing manual and automatic analysis protocols. Upon nucleotide binding, extensive chemical shift perturbations could be observed. These data provide evidence for a symmetric DGK trimer with all of its three active sites concurrently occupied. Additionally, we could detect that the nucleotide substrate induces a substantial conformational change, most likely directing DGK into its catalytic active form. Furthermore, functionally relevant interprotomer interactions are identified by DNP-enhanced MAS NMR in combination with site-directed mutagenesis and functional assays.
miRNA analysis with Prost! reveals evolutionary conservation of organ-enriched expression and post-transcriptional modifications in three-spined stickleback and zebrafish
Authors: Desvignes Thomas , Batzel Peter , Sydes Jason , Eames B. Frank , Postlethwait John H.
Published: March 2019

MicroRNAs (miRNAs) can have organ-specific expression and functions; they can originate from dedicated miRNA genes, from non-canonical miRNA genes, or from mirror-miRNA genes and can also experience post-transcriptional variation. It remains unclear, however, which mechanisms of miRNA production or modification are organ-specific and the extent of their evolutionary conservation. To address these issues, we developed the software Prost! (PRocessing Of Short Transcripts), which, among other features, helps quantify mature miRNAs, accounts for post-transcriptional processing, such as nucleotide editing, and identifies mirror-miRNAs. Here, we applied Prost! to annotate and analyze miRNAs in three-spined stickleback (Gasterosteus aculeatus), a model fish for evolutionary biology reported to have a miRNome larger than most teleost fish. Zebrafish (Danio rerio), a distantly related teleost with a well-known miRNome, served as comparator. Our results provided evidence for the existence of 286 miRNA genes and 382 unique mature miRNAs (excluding mir430 gene duplicates and the vaultRNA-derived mir733), which doesn’t represent a miRNAome larger than other teleost miRNomes. In addition, small RNA sequencing data from brain, heart, testis, and ovary in both stickleback and zebrafish identified suites of mature miRNAs that display organ-specific enrichment, many of which are evolutionarily-conserved in the brain and heart in both species. These data also supported the hypothesis that evolutionarily-conserved, organ-specific mechanisms may regulate post-transcriptional variations in miRNA sequence. In both stickleback and zebrafish, miR2188-5p was edited frequently with similar nucleotide changes in the seed sequence with organ specific editing rates, highest in the brain. In summary, Prost! is a new tool to identify and understand small RNAs, to help clarify a species’ miRNA biology as shown here for an important model for the evolution of developmental mechanisms, and to provide insight into organ-enriched expression and the evolutionary conservation of miRNA post-transcriptional modifications.
Physicochemical, cytotoxicity and in vivo biocompatibility of a high-plasticity calcium-silicate based material
Authors: Ferreira Cláudio M. A. , Sassone Luciana M. , Gonçalves Alexia S. , de Carvalho Jorge José , Tomás-Catalá Christopher J. , García-Bernal David , Oñate-Sánchez Ricardo E. , Rodríguez-Lozano Francisco J. , Silva Emmanuel João Nogueira Leal
Published: March 2019

The purpose of this work was to evaluate the physicochemical properties, the cytotoxicity and in vivo biocompatibility of MTA Repair HP (MTA HP) and White MTA (WMTA). The setting time, flow, radiopacity and water solubility were assessed. To the cytotoxicity assay, primary human osteoblast cells were exposed to several dilutions of both materials eluates. MTT assay, apoptosis assay and cell adhesion assay were performed. The in vivo biocompatibility was evaluated through histological analysis using different staining techniques. No differences were observed between MTA HP and WMTA for setting time, radiopacity, solubility and water absorption (P > 0.05). However, MTA HP showed a significantly higher flow when compared to WMTA (P < 0.05). Cell viability results revealed that the extracts of WMTA and MTA HP promoted the viability of osteoblasts. After incubation of cells with the endodontic cement extracts, the percentage of apoptotic or necrotic cells was very low (<3%). Furthermore, SEM results showed a high degree of cell proliferation and adhesion on both groups. MTA HP showed similar in vivo biocompatibility to the WMTA and the control group in all time-points. The MTA HP presented adequate physicochemical and biological properties with improved flow ability when compared to WMTA. Such improved flow ability may be a result of the addition of a plasticizing agent and should be related to an improvement in the handling of MTA HP.
Impact of Wuyiencin Application on the Soil Microbial Community and Fate of Typical Antibiotic Resistance Genes
Authors: Shi Liming , Ge Beibei , Liu Binghua , Liu Xingang , Jiang Mingguo , Zhang Kecheng
Published: March 2019

Antibiotic resistance genes (ARGs) have raised numerous concerns in recent years as emerging environmental contaminants. At present, research on environmental contamination by antibiotics focuses on medical, animal husbandry, and aquaculture fields, with few studies on environmental contamination by agricultural antibiotics in the field of plant protection. Wuyiencin is a low toxicity, high efficiency, and broad-spectrum agricultural antibiotic. It has been widely used in agricultural production and it effectively controls crop fungal diseases. In the present study, pot experiments with four soil treatments (A, B, C and D) were set up in a greenhouse to investigate the effect of the application of wuyiencin on the fate of typical ARGs and microbial community. Eight typical ARGs were detected by real-time PCR and the microbial communities were analyzed using high-throughput sequencing. The results showed that wuyiencin neither significantly influenced ARG abundance and absolute gene copy numbers, nor significantly varied microbial community among treatments. Since it only was short-term results, and the detection number of ARGs was limited, whether wuyiencin is safe or not to ecological environment when using for long-term will need further deep research.
A Universal Stamping Method of Graphene Transfer for Conducting Flexible and Transparent Polymers
Authors: Chandrashekar Bananakere Nanjegowda , Smitha Ankanahalli Shankaregowda , Wu Yingchun , Cai Nianduo , Li Yunlong , Huang Ziyu , Wang Weijun , Shi Run , Wang Jingwei , Liu Shiyuan , Krishnaveni S. , Wang Fei , Cheng Chun
Published: March 2019

Transfer method of chemically vapor deposition graphene is an appealing issue to realize its application as flexible and transparent electrodes. A universal stamping method to transfer as grown graphene from copper onto different flexible and transparent polymers (FTPs) reported here ensures simple, robust, rapid, clean and low-cost. This method relies on coating ethylene vinyl acetate (EVA) onto the as grown graphene, binding EVA coated graphene/Cu with FTPs and delamination by hydrogen bubbling process, which is analogous to the method used by stamping process where ink carries the imprint of the object onto any materials. The fate of the stamping method depends on how strongly the adhesion of EVA coated graphene/Cu with target FTPs. Interestingly, we have found that the thin film of EVA/graphene/Cu can only bind strongly with the FTPs of less than 25 µm in thickness and lower glass transition temperature value to the EVA while wide range of other FTPs are considered upon surface engineering to enhance the binding strength between FTPs and EVA. What’s more, the electrical performance was investigated with a demonstration of triboelectric nanogenerators which confirmed the reliability of graphene transfer onto the FTPs and prospect for the development of flexible and transparent electronics.
Collective influencers in protein interaction networks
Authors: Boltz T. A. , Devkota P. , Wuchty Stefan
Published: March 2019

Recent research increasingly shows the relevance of network based approaches for our understanding of biological systems. Analyzing human protein interaction networks, we determined collective influencers (CI), defined as network nodes that damage the integrity of the underlying networks to the utmost degree. We found that CI proteins were enriched with essential, regulatory, signaling and disease genes as well as drug targets, indicating their biological significance. Also by focusing on different organisms, we found that CI proteins had a penchant to be evolutionarily conserved as CI proteins, indicating the fundamental role that collective influencers in protein interaction networks plays for our understanding of regulation, diseases and evolution.
Structure guided design of an antibacterial peptide that targets UDP-N-acetylglucosamine acyltransferase
Authors: Dangkulwanich Manchuta , Raetz Christian R. H. , Williams Allison H.
Published: March 2019

UDP-N-acetylglucosamine (UDP-GlcNAc) acyltransferase (LpxA) catalyzes the first step of lipid A biosynthesis, the transfer of anR-3-hydroxyacyl chain from its acyl carrier protein (ACP) to the 3-OH group of UDP-GlcNAc. Essential in the growth of Gram-negative bacteria, LpxA is a logical target for antibiotics design. A pentadecapeptide (Peptide 920) with high affinity towards LpxA was previously identified in a phage display library. Here we created a small library of systematically designed peptides with the length of four to thirteen amino acids using Peptide 920 as a scaffold. The concentrations of these peptides at which 50% of LpxA is inhibited (IC50) range from 50 nM to >100 μM. We determined the crystal structure ofE. coliLpxA in a complex with a potent inhibitor. LpxA-inhibitor interaction, solvent model and all contributing factors to inhibitor efficacy were well resolved. The peptide primarily occludes the ACP binding site of LpxA. Interactions between LpxA and the inhibitor are different from those in the structure of Peptide 920. The inhibitory peptide library and the crystal structure of inhibitor-bound LpxA described here may further assist in the rational design of inhibitors with antimicrobial activity that target LpxA and potentially other acyltransferases.
Mathematical modelling indicates that lower activity of the haemostatic system in neonates is primarily due to lower prothrombin concentration
Authors: Siekmann Ivo , Bjelosevic Stefan , Landman Kerry , Monagle Paul , Ignjatovic Vera , Crampin Edmund J.
Published: March 2019

Haemostasis is governed by a highly complex system of interacting proteins. Due to the central role of thrombin, thrombin generation and specifically the thrombin generation curve (TGC) is commonly used as an indicator of haemostatic activity. Functional characteristics of the haemostatic system in neonates and children are significantly different compared with adults; at the same time plasma levels of haemostatic proteins vary considerably with age. However, relating one to the other has been difficult, both due to significant inter-individual differences for individuals of similar age and the complexity of the biochemical reactions underlying haemostasis. Mathematical modelling has been very successful at representing the biochemistry of blood clotting. In this study we address the challenge of large inter-individual variability by parameterising the Hockin-Mann model with data from individual patients, across different age groups from neonates to adults. Calculating TGCs for each patient of a specific age group provides us with insight into the variability of haemostatic activity across that age group. From our model we observe that two commonly used metrics for haemostatic activity are significantly lower in neonates than in older patients. Because both metrics are strongly determined by prothrombin and prothrombin levels are considerably lower in neonates we conclude that decreased haemostatic activity in neonates is due to lower prothrombin availability.
Infrared-sensing snakes select ambush orientation based on thermal backgrounds
Authors: Schraft Hannes A. , Bakken George S. , Clark Rulon W.
Published: March 2019

Sensory information drives the ecology and behaviour of animals, and some animals are able to detect environmental cues unavailable to us. For example, rattlesnakes use infrared (IR) radiation to detect warm prey at night when visual cues are reduced. Until recently these sensory worlds have been inaccessible to human observers; now technology can allow us to “eavesdrop” on these species and understand how sensory perception drives ecology and behaviour. We used thermography and computer simulations to examine how prey-background temperature contrast and areas of temperature transitions influence the angular orientation of free-ranging rattlesnakes once they have selected an ambush site. We tracked free-ranging sidewinder rattlesnakes Crotalus cerastes to their selected ambush sites and recorded 360° near-ground thermographic panoramas from the centre of the ambush site. A computer simulation then moved a simulated prey item across the panorama and computed a contrast index for all directions. Rattlesnakes did not face ambush directions that offered stronger contrast than average, but they demonstrated a striking tendency to face directions with strong thermal transitions. Background transitions likely create a readily detected, rapidly changing stimulus when a prey animal passes. Quantifications of sensory environments like this one can boost our comprehension of how sensory function impacts the ecology, behaviour, and evolution of animals.