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Tailoring cell–surface interactions is important for the of design medical implants as well as regenerative medicine and tissue engineering materials. Here the single parameter system is transcended via translating hard nanotopology into soft polymeric hydrogel structures via hydrogel imprinting lithography. The response of these cells to the nanotopology of the same dimensions but with different mechanical properties displays unexpected behavior between “hard” tissue cells and “soft” tissue cells.
The Puna Plateau, adjacent Eastern Cordillera and the Sierras Pampeanas of the central Andes are largely characterized by thick-skinned, basement-involved deformation. The Puna Plateau hosts similar to N-S trending bedrock ranges bounded by deep-seated reverse faults and sedimentary basins. We contribute to the understanding of thick-skinned dynamics in the Puna Plateau by constraining regional kinematics of the poorly understood southern Puna Plateau through a multidisciplinary approach. On the southeastern plateau, sandstone modal composition and detrital zircon U-Pb and apatite fission-track data from Cenozoic strata indicate basin accumulation during the late Eocene to early Oligocene (similar to 38-28 Ma). Provenance analysis reveals the existence of a regional-scale basin covering the southern Puna Plateau during late Eocene to early Oligocene time (similar to 38-28 Ma) that was sourced from both the western plateau and the eastern plateau margin and had a depocenter located to the west. Petrographic and detrital zircon U-Pb data reveal erosion of proximal western and eastern sources after 12 Ma, in mid-late Miocene time. This indicates that the regional basin was compartmentalized into small-scale depocenters by the growth of basement-cored ranges continuing into the late Miocene (similar to 12-8 Ma). We suggest that the Cenozoic history of the southern Puna Plateau records the formation of a regional basin that was possibly driven by lithospheric flexure during the late Eocene to early Oligocene, before the growth of distributed basement-cored ranges starting as early as the late Oligocene. (C) 2015 Elsevier B.V. All rights reserved.
The physiological functions of sphingolipids in animals have been intensively studied, while less attention has been paid to their roles in plants. Here, we reveal the involvement of sphingolipid delta8 desaturase (SlSLD) in the chilling resistance of tomato (Solanum lycopersicum cv. Micro-Tom). We used the virus-induced gene silencing (VIGS) approach to knock-down SlSLD expression in tomato leaves, and then evaluated chilling resistance. Changes in leaf cell structure under a chilling treatment were observed by transmission electron microscopy. In control plants, SlSLD was highly expressed in the fruit and leaves in response to a chilling treatment. The degree of chilling damage was greater in SlSLD-silenced plants than in control plants, indicating that SlSLD knock-down significantly reduced the chilling resistance of tomato. Compared with control plants, SlSLD-silenced plants showed higher relative electrolytic leakage and malondialdehyde content, and lower superoxide dismutase and peroxidase activities after a chilling treatment. Chilling severely damaged the chloroplasts in SlSLD-silenced plants, resulting in the disruption of chloroplast membranes, swelling of thylakoids, and reduced granal stacking. Together, these results show that SlSLD is crucial for chilling resistance in tomato.
The all-female Amazon molly (Poecilia formosa) originated from a single hybridization of two bisexual ancestors, Atlantic molly (Poecilia mexicana) and sailfin molly (Poecilia latipinna). As a gynogenetic species, the Amazon molly needs to copulate with a heterospecific male, but the genetic information of the sperm-donor does not contribute to the next generation, as the sperm only acts as the trigger for the diploid eggs’ embryogenesis. Here, we study the sequence evolution and gene expression of the duplicated genes coding for androgen receptors (ars) and other pathway-related genes, i.e., the estrogen receptors (ers) and cytochrome P450, family19, subfamily A, aromatase genes (cyp19as), in the Amazon molly, in comparison to its bisexual ancestors. Mollies possess–as most other teleost fish—two copies of the ar, er, and cyp19a genes, i.e., arα/arβ, erα/erβ1, and cyp19a1 (also referred as cyp19a1a)/cyp19a2 (also referred to as cyp19a1b), respectively. Non-synonymous single nucleotide polymorphisms (SNPs) among the ancestral bisexual species were generally predicted not to alter protein function. Some derived substitutions in the P. mexicana and one in P. formosa are predicted to impact protein function. We also describe the gene expression pattern of the ars and pathway-related genes in various tissues (i.e., brain, gill, and ovary) and provide SNP markers for allele specific expression research. As a general tendency, the levels of gene expression were lowest in gill and highest in ovarian tissues, while expression levels in the brain were intermediate in most cases. Expression levels in P. formosa were conserved where expression did not differ between the two bisexual ancestors. In those cases where gene expression levels significantly differed between the bisexual species, P. formosa expression was always comparable to the higher expression level among the two ancestors. Interestingly, erβ1 was expressed neither in brain nor in gill in the analyzed three molly species, which implies a more important role of erα in the estradiol synthesis pathway in these tissues. Furthermore, our data suggest that interactions of steroid-signaling pathway genes differ across tissues, in particular the interactions of ars and cyp19as.
The all-female Amazon molly (Poecilia formosa) originated from a single hybridization of two bisexual ancestors, Atlantic molly (Poecilia mexicana) and sailfin molly (Poecilia latipinna). As a gynogenetic species, the Amazon molly needs to copulate with a heterospecific male, but the genetic information of the sperm-donor does not contribute to the next generation, as the sperm only acts as the trigger for the diploid eggs’ embryogenesis. Here, we study the sequence evolution and gene expression of the duplicated genes coding for androgen receptors (ars) and other pathway-related genes, i.e., the estrogen receptors (ers) and cytochrome P450, family19, subfamily A, aromatase genes (cyp19as), in the Amazon molly, in comparison to its bisexual ancestors. Mollies possess–as most other teleost fish—two copies of the ar, er, and cyp19a genes, i.e., arα/arβ, erα/erβ1, and cyp19a1 (also referred as cyp19a1a)/cyp19a2 (also referred to as cyp19a1b), respectively. Non-synonymous single nucleotide polymorphisms (SNPs) among the ancestral bisexual species were generally predicted not to alter protein function. Some derived substitutions in the P. mexicana and one in P. formosa are predicted to impact protein function. We also describe the gene expression pattern of the ars and pathway-related genes in various tissues (i.e., brain, gill, and ovary) and provide SNP markers for allele specific expression research. As a general tendency, the levels of gene expression were lowest in gill and highest in ovarian tissues, while expression levels in the brain were intermediate in most cases. Expression levels in P. formosa were conserved where expression did not differ between the two bisexual ancestors. In those cases where gene expression levels significantly differed between the bisexual species, P. formosa expression was always comparable to the higher expression level among the two ancestors. Interestingly, erβ1 was expressed neither in brain nor in gill in the analyzed three molly species, which implies a more important role of erα in the estradiol synthesis pathway in these tissues. Furthermore, our data suggest that interactions of steroid-signaling pathway genes differ across tissues, in particular the interactions of ars and cyp19as.
QuestionHow important is the effect of micro-relief and vegetation type on the characteristics of vascular plants and bryophytes in low-centred polygons? LocationSiberian Arctic, Russia. MethodsEight low-centred polygons in northern Siberia were surveyed for vegetation along transects running from the rim to the pond via the rim-pond transition of each polygon and across a vegetation type gradient from open forest to tundra. ResultsThe cover of vascular plants and bryophytes displays no significant differences between the rim and rim-pond transition but is significantly lower in the pond section of the polygons. Alpha-diversity of vascular plants decreases strongly from rim to pond, whereas bryophyte diversity in pond plots is significantly distinct from the rim and the rim-pond transition. There is no clear trend in cover for either plant group along the vegetation type transect and only a weak trend in -diversity. However, both gradients are reflected in the compositional turnover. The applied indicator species analysis identified taxa characteristic of certain environmental conditions. Among others, we found vascular plants primarily characteristic of the rim and bryophyte taxa characteristic of each micro-relief level and vegetation type. ConclusionsThe observed gradual pattern in -diversity and composition of polygonal vegetation suggests that micro-relief is the main driver of changes in the vegetation composition, while vegetation type and the related forest cover change are of subordinate importance for polygonal vegetation patterns along the Siberian tree line.
Iceland is located on the Mid-Atlantic Ridge which is the plate boundary between the Eurasian and the North American plates. It is one of the few places on earth where an active spreading centre is located onshore but the stress pattern has not been extensively investigated so far. In this paper we present a comprehensive compilation of the orientation of maximum horizontal stress (S-Hmax). In particular we interpret borehole breakouts and drilling induced fractures from borehole image logs in 57 geothermal wells onshore Iceland. The borehole results are combined with other stress indicators including earthquake focal mechanism solutions, geological information and overcoring measurements resulting in a dataset with 495 data records for the S-Hmax orientation. The reliability of each indicator is assessed according to the quality criteria of the World Stress Map project The majority of S-Hmax orientation data records in Iceland is derived from earthquake focal mechanism solutions (35%) and geological fault slip inversions (26%). 20% of the data are borehole related stress indicators. In addition minor shares of S-Hmax orientations are compiled, amongst others, from focal mechanism inversions and the alignment of fissure eruptions. The results show that the S-Hmax orientations derived from different depths and stress indicators are consistent with each other.
The resulting pattern of the present-day stress in Iceland has four distinct subsets of S-Hmax orientations. The S-Hmax orientation is parallel to the rift axes in the vicinity of the active spreading regions. It changes from NE-SW in the South to approximately N-S in central Iceland and NNW-SSE in the North. In the Westfjords which is located far away from the ridge the regional S-Hmax rotates and is parallel to the plate motion. (C) 2016 Elsevier B.V. All rights reserved.
estimating mean throughfall
(2016)
The selection of an appropriate spatial extent of a sampling plot is one among several important decisions involved in planning a throughfall sampling scheme. In fact, the choice of the extent may determine whether or not a study can adequately characterize the hydrological fluxes of the studied ecosystem. Previous attempts to optimize throughfall sampling schemes focused on the selection of an appropriate sample size, support, and sampling design, while comparatively little attention has been given to the role of the extent. In this contribution, we investigated the influence of the extent on the representativeness of mean throughfall estimates for three forest ecosystems of varying stand structure. Our study is based on virtual sampling of simulated throughfall fields. We derived these fields from throughfall data sampled in a simply structured forest (young tropical forest) and two heterogeneous forests (old tropical forest, unmanaged mixed European beech forest). We then sampled the simulated throughfall fields with three common extents and various sample sizes for a range of events and for accumulated data. Our findings suggest that the size of the study area should be carefully adapted to the complexity of the system under study and to the required temporal resolution of the throughfall data (i.e. event-based versus accumulated). Generally, event-based sampling in complex structured forests (conditions that favor comparatively long autocorrelations in throughfall) requires the largest extents. For event-based sampling, the choice of an appropriate extent can be as important as using an adequate sample size. (C) 2016 Elsevier B.V. All rights reserved.
In November 2015, the 14th Session of the Assembly of States Parties to the Rome Statute of the International Criminal Court (ICC) adopted, by consensus, an amendment providing for the deletion of Article 124 of the ICC Statute, which so far enables contracting parties, when joining the Statute, to opt out from the ICC’s treaty-based war crimes-related jurisdiction. After considering the genesis of the provision and the practice arising under Article 124 of the ICC Statute so far, this article considers the arguments for and against the deletion of Article 124 in light of the increasingly small number of accessions to the ICC Statute that have been forthcoming in the last few years. It also analyses the quite strict requirements for the entry into force of the amendment, as well as the effect of the entry into force of the amendment on possible declarations having been made pending such entry into force. It ends by considering the positive effect a continued applicability of Article 124 may have on states so far being reluctant to accede to the ICC Statute.
In November 2015, the 14th Session of the Assembly of States Parties to the Rome Statute of the International Criminal Court (ICC) adopted, by consensus, an amendment providing for the deletion of Article 124 of the ICC Statute, which so far enables contracting parties, when joining the Statute, to opt out from the ICC’s treaty-based war crimes-related jurisdiction. After considering the genesis of the provision and the practice arising under Article 124 of the ICC Statute so far, this article considers the arguments for and against the deletion of Article 124 in light of the increasingly small number of accessions to the ICC Statute that have been forthcoming in the last few years. It also analyses the quite strict requirements for the entry into force of the amendment, as well as the effect of the entry into force of the amendment on possible declarations having been made pending such entry into force. It ends by considering the positive effect a continued applicability of Article 124 may have on states so far being reluctant to accede to the ICC Statute.
Article 15bis: Exercise of jurisdiction over the crime of aggression (State referal, proprio motu)
(2016)
Predicate focus
(2016)
Background: Cannabis is the most commonly used illegal substance among adolescents and young adults. Problematic cannabis use is often associated with comorbid psychopathological problems. The purpose of the current study was to elucidate the underlying developmental processes connecting externalizing and internalizing psychopathology in childhood and adolescence with problematic cannabis use in young adulthood. Methods: Data were drawn from the Mannheim Study of Children at Risk, an ongoing epidemiological cohort study from birth to adulthood. For n = 307 participants, symptom scores of conduct/oppositional defiant disorder, attention problems, hyperactivity/impulsivity, and internalizing disorders were available for the periods of childhood (4.5-11 years) and adolescence (15 years). At age 25 years, problematic cannabis use was assessed via clinical interview and a self-rating questionnaire. Results: At age 25 years, problematic cannabis use was identified in n = 28 participants (9.1%). Childhood conduct/oppositional behavior problems were predictive of problematic cannabis use during young adulthood when comorbid symptoms were controlled for. No such effect was found for childhood attention, hyperactivity/impulsivity or internalizing problems. With respect to psychopathological symptoms during adolescence, only attention problems were significantly related to later problematic cannabis use when controlling for comorbidity. Conclusions: The current study highlights the role of conduct/oppositional behavior problems during childhood and attention problems during adolescence in later problematic cannabis use. It sheds more light on the developmental sequence of childhood and adolescence psychopathology and young adult cannabis use, which is a prerequisite for effective prevention approaches. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
Unlike for other retroviruses, only a few host cell factors that aid the replication of foamy viruses (FVs) via interaction with viral structural components are known. Using a yeast-two-hybrid (Y2H) screen with prototype FV (PFV) Gag protein as bait we identified human polo-like kinase 2 (hPLK2), a member of cell cycle regulatory kinases, as a new interactor of PFV capsids. Further Y2H studies confirmed interaction of PFV Gag with several PLKs of both human and rat origin. A consensus Ser-Thr/Ser-Pro (S-T/S-P) motif in Gag, which is conserved among primate FVs and phosphorylated in PFV virions, was essential for recognition by PLKs. In the case of rat PLK2, functional kinase and polo-box domains were required for interaction with PFV Gag. Fluorescently-tagged PFV Gag, through its chromatin tethering function, selectively relocalized ectopically expressed eGFP-tagged PLK proteins to mitotic chromosomes in a Gag STP motif-dependent manner, confirming a specific and dominant nature of the Gag-PLK interaction in mammalian cells. The functional relevance of the Gag-PLK interaction was examined in the context of replication-competent FVs and single-round PFV vectors. Although STP motif mutated viruses displayed wild type (wt) particle release, RNA packaging and intra-particle reverse transcription, their replication capacity was decreased 3-fold in single-cycle infections, and up to 20-fold in spreading infections over an extended time period. Strikingly similar defects were observed when cells infected with single-round wt Gag PFV vectors were treated with a pan PLK inhibitor. Analysis of entry kinetics of the mutant viruses indicated a post-fusion defect resulting in delayed and reduced integration, which was accompanied with an enhanced preference to integrate into heterochromatin. We conclude that interaction between PFV Gag and cellular PLK proteins is important for early replication steps of PFV within host cells.
Alternaria (A.) is a genus of widespread fungi capable of producing numerous, possibly health-endangering Alternaria toxins (ATs), which are usually not the focus of attention. The formation of ATs depends on the species and complex interactions of various environmental factors and is not fully understood. In this study the influence of temperature (7 degrees C, 25 degrees C), substrate (rice, wheat kernels) and incubation time (4, 7, and 14 days) on the production of thirteen ATs and three sulfoconjugated ATs by three different Alternaria isolates from the species groups A. tenuissima and A. infectoria was determined. High-performance liquid chromatography coupled with tandem mass spectrometry was used for quantification. Under nearly all conditions, tenuazonic acid was the most extensively produced toxin. At 25 degrees C and with increasing incubation time all toxins were formed in high amounts by the two A. tenuissima strains on both substrates with comparable mycotoxin profiles. However, for some of the toxins, stagnation or a decrease in production was observed from day 7 to 14. As opposed to the A. tenuissima strains, the A. infectoria strain only produced low amounts of ATs, but high concentrations of stemphyltoxin III. The results provide an essential insight into the quantitative in vitro AT formation under different environmental conditions, potentially transferable to different field and storage conditions.
The Groningen gas field serves as a natural laboratory for production-induced earthquakes, because no earthquakes were observed before the beginning of gas production. Increasing gas production rates resulted in growing earthquake activity and eventually in the occurrence of the 2012M(w) 3.6 Huizinge earthquake. At least since this event, a detailed seismic hazard and risk assessment including estimation of the maximum earthquake magnitude is considered to be necessary to decide on the future gas production. In this short note, we first apply state-of-the-art methods of mathematical statistics to derive confidence intervals for the maximum possible earthquake magnitude m(max). Second, we calculate the maximum expected magnitude M-T in the time between 2016 and 2024 for three assumed gas-production scenarios. Using broadly accepted physical assumptions and 90% confidence level, we suggest a value of m(max) 4.4, whereas M-T varies between 3.9 and 4.3, depending on the production scenario.
In the present study, we summarize and evaluate the endeavors from recent years to estimate the maximum possible earthquake magnitude m(max) from observed data. In particular, we use basic and physically motivated assumptions to identify best cases and worst cases in terms of lowest and highest degree of uncertainty of m(max). In a general framework, we demonstrate that earthquake data and earthquake proxy data recorded in a fault zone provide almost no information about m(max) unless reliable and homogeneous data of a long time interval, including several earthquakes with magnitude close to m(max), are available. Even if detailed earthquake information from some centuries including historic and paleoearthquakes are given, only very few, namely the largest events, will contribute at all to the estimation of m(max), and this results in unacceptably high uncertainties. As a consequence, estimators of m(max) in a fault zone, which are based solely on earthquake-related information from this region, have to be dismissed.
The combination of high-performance liquid chromatography and electrospray ionization ion mobility spectrometry facilitates the two-dimensional separation of complex mixtures in the retention and drift time plane. The ion mobility spectrometer presented here was optimized for flow rates customarily used in high-performance liquid chromatography between 100 and 1500 mu L/min. The characterization of the system with respect to such parameters as the peak capacity of each time dimension and of the 2D spectrum was carried out based on a separation of a pesticide mixture containing 24 substances. While the total ion current chromatogram is coarsely resolved, exhibiting coelutions for a number of compounds, all substances can be separately detected in the 2D plane due to the orthogonality of the separations in retention and drift dimensions. Another major advantage of the ion mobility detector is the identification of substances based on their characteristic mobilities. Electrospray ionization allows the detection of substances lacking a chromophore. As an example, the separation of a mixture of 18 amino acids is presented. A software built upon the free mass spectrometry package OpenMS was developed for processing the extensive 2D data. The different processing steps are implemented as separate modules which can be arranged in a graphic workflow facilitating automated processing of data.
Measures for interoperability of phenotypic data: minimum information requirements and formatting
(2016)
Background: Plant phenotypic data shrouds a wealth of information which, when accurately analysed and linked to other data types, brings to light the knowledge about the mechanisms of life. As phenotyping is a field of research comprising manifold, diverse and time-consuming experiments, the findings can be fostered by reusing and combining existing datasets. Their correct interpretation, and thus replicability, comparability and interoperability, is possible provided that the collected observations are equipped with an adequate set of metadata. So far there have been no common standards governing phenotypic data description, which hampered data exchange and reuse. Results: In this paper we propose the guidelines for proper handling of the information about plant phenotyping experiments, in terms of both the recommended content of the description and its formatting. We provide a document called "Minimum Information About a Plant Phenotyping Experiment", which specifies what information about each experiment should be given, and a Phenotyping Configuration for the ISA-Tab format, which allows to practically organise this information within a dataset. We provide examples of ISA-Tab-formatted phenotypic data, and a general description of a few systems where the recommendations have been implemented. Conclusions: Acceptance of the rules described in this paper by the plant phenotyping community will help to achieve findable, accessible, interoperable and reusable data.
The Proteasome Acts as a Hub for Plant Immunity and Is Targeted by Pseudomonas Type III Effectors
(2016)
Recent evidence suggests that the ubiquitin-proteasome system is involved in several aspects of plant immunity and that a range of plant pathogens subvert the ubiquitin-proteasome system to enhance their virulence. Here, we show that proteasome activity is strongly induced during basal defense in Arabidopsis (Arabidopsis thaliana). Mutant lines of the proteasome subunits RPT2a and RPN12a support increased bacterial growth of virulent Pseudomonas syringae pv tomato DC3000 (Pst) and Pseudomonas syringae pv maculicola ES4326. Both proteasome subunits are required for pathogen-associated molecular pattern-triggered immunity responses. Analysis of bacterial growth after a secondary infection of systemic leaves revealed that the establishment of systemic acquired resistance (SAR) is impaired in proteasome mutants, suggesting that the proteasome also plays an important role in defense priming and SAR. In addition, we show that Pst inhibits proteasome activity in a type III secretion-dependent manner. A screen for type III effector proteins from Pst for their ability to interfere with proteasome activity revealed HopM1, HopAO1, HopA1, and HopG1 as putative proteasome inhibitors. Biochemical characterization of HopM1 by mass spectrometry indicates that HopM1 interacts with several E3 ubiquitin ligases and proteasome subunits. This supports the hypothesis that HopM1 associates with the proteasome, leading to its inhibition. Thus, the proteasome is an essential component of pathogen-associated molecular pattern-triggered immunity and SAR, which is targeted by multiple bacterial effectors.
Since spring 2014 the relations between the EU and Russia are stuck in an Ice Age. From a Western point of view, especially the annexation of Crimea by Russia and the intervention in the conflict in Ukraine are responsible. The EU has frozen their relations to Russia and applied sanctions against it. Russia reacted in the same way. Can this vicious circle be broken without betraying the values of the EU? This book presents the analysis and ideas of social scientists from Germany, Poland and Russia. The reasons for this crisis are seen quite differently but all try to find a way out of the current confrontation.
This book offers a clear, critical, and comprehensive overview of theoretical and experimental work on information structure. Different chapters examine the main theories of information structure in syntax, phonology, and semantics as well as perspectives from psycholinguistics and other relevant fields. Following the editors’ introduction the book is divided into four parts. The first, on theories of and theoretical perspectives on information structure, includes chapters on topic, prosody, and implicature. Part II covers a range of current issues in the field, including focus, quantification, and sign languages, while Part III is concerned with experimental approaches to information structure, including processes involved in its acquisition and comprehension. The final part contains a series of linguistic case studies drawn from a wide variety of the world’s language families
This edited volume provides insight into how digital badges may enhance formal and informal education by focusing on technical design issues including organizational requirements, instructional design, and deployment. It features current research exploring the theoretical foundation and empirical evidence of the utilization of digital badges as well as case studies that describe current practices and experiences in the use of digital badges for motivation, learning, and instruction in K-12, higher education, workplace learning, and further education settings.
The law of treaties; or, should this book exist? / Vaughan LoweThe law of treaties through the interplay of its different sources / Enzo Cannizzaro -- Regulating treaties: a comparative perspective / Martins Paparinskis -- Theorizing treaties: the consequences of the contractual analogy / Akbar Rasulov -- The effects of treaties in domestic law / André Nollkaemper -- The temporal dimension: non-retroactivity and its discontents / Markus Kotzur -- The spatial dimension: treaties and territory / Marko Milanovic -- The personal dimension: challenges to the pacta tertiis rule / Alexander Proelss -- Formalism versus flexibility in the law of treaties / Jean d'Aspremont -- Integrity versus flexibility in the application of treaties / Katherine del Mar -- Pacta sunt servanda versus flexibility in the suspension and termination of treaties / Sotirios-Ioannis Lekkas and Antonios Tzanakopoulos -- Uniformity versus specialisation (1): the quest for a uniform law of inter-state treaties / Malgosia Fitzmaurice and Panos Merkouris -- Uniformity versus specialisation (2): a uniform regime of treaty interpretation? / Michael Waibel -- Regime-collisions: tensions between treaties (and how to solve them) / Jasper Finke -- Responding to deliberately-created treaty conflicts / Surabhi Ranganathan -- Treaty breaches and responses / Christian J Tams -- Succession to treaties and the inherent limits of international law / Andreas Zimmermann and James G. Devaney -- Treaties and armed conflict / Yael Ronen -- Treaties and international organisations: uneasy analogies / Philippa Webb -- Treaty law and multinational enterprises: more than internationalized contracts? / Markos Karavias -- Treaties and individuals: of beneficiaries, duty-bearers, users, and participants / Ilias Plakokefalos