bluuebunny/crossref_metadata_2025 huggingface dataset:
Strontium isotopes and mobility of a Columbian mammoth (Mammuthus columbi) population, Laguna de las Cruces, San Luis Potosí, México
Authors: PÉREZ-CRESPO VÍCTOR ADRIÁN , SCHAAF PETER , SOLÍS-PICHARDO GABRIELA , ARROYO-CABRALES JOAQUÍN , ALVA-VALDIVIA LUIS M. , TORRES-HERNÁNDEZ JOSÉ RAMÓN
Published: February 2016

By using strontium isotopic ratios of dental enamel from molars, we were able to reconstruct the migration context for three individuals of a Columbian mammoth population (Mammuthus columbi) around Laguna de las Cruces, San Luis Potosí, central México. A three-step leaching procedure was applied to eliminate secondary Sr contributions in the molar enamel. One of the studied individuals showed87Sr/86Sr ratios similar to those obtained from soils and plants from Laguna de las Cruces and was identified as local, whereas the other two mammoths had different molar87Sr/86Sr values, indicative of migration and mobility contexts.
The occurrence of the filarial nematodeDirofilaria repensin canine hosts from Maio Island, Cape Verde
Authors: Marcos R. , Pereira C. , Maia J.P. , Santos M. , Luzzago C. , Lauzi S. , Genchi C. , Faustino A. , Puente-Payo P.
Published: February 2016

The prevalence of canineDirofilariainfection in Maio Island (Cape Verde) was analysed by serology, morphological and molecular identification of the parasite species. Blood and sera were collected from 150 dogs and 80 cats aged over 6 months from various localities of the island. DNA was extracted from blood and samples were screened by polymerase chain reaction (PCR) using microfilaria-specific primers. NoDirofilaria immitiswas found in dogs whileD. repensmicrofilariae were found in 5.3% of dogs and 6% were positive by PCR. The species identity was confirmed by sequencing of PCR products, which showed almost 100% homology withD. repensEuropean sequences published in GenBank. No difference inDirofilariainfection was observed between males and females or in dogs with different weights. However, older dogs and those from the western part of Maio Island were more frequently infected. NoDirofilariawas found in cats. This study represents the first evidence ofD. repensin Cape Verde (West Africa) and highlights the need for implementing control measures and for a better surveillance of dirofilariosis in Africa.
Estimates of cetacean by-catch in the turbot fishery on the Turkish Western Black Sea Coast in 2007 and 2008
Authors: Tonay Arda M.
Published: February 2016

This is the first study estimating cetacean by-catch in the Turkish western Black Sea turbot fishery. One turbot fishing boat was observed during two fishing seasons, from April through July 2007 and April through mid-September 2008. During this time, 24 harbour porpoises and one bottlenose dolphin were caught in turbot trammel nets. The by-catch rate was found to be 0.18 for harbour porpoise and 0.01 for bottlenose dolphin individuals per kilometre in 2007, and 0.19 for harbour porpoise individuals in 2008. It is estimated that the total numbers of harbour porpoises killed in the Turkish western Black Sea during the legal fishing period (April and July) were 167 ± 153 (CV: 0.92) in 2007 and 329 ± 220 (CV: 0.67) in 2008, and the number killed during both legal and illegal periods of turbot fishing were 2011 ± 742 (CV: 0.37) in 2007 and 2294 ± 806 (CV: 0.35) in 2008. The estimated range of harbour porpoise by-catch in the turbot fishery on the Turkish western Black Sea coast is between these two estimates. The by-caught harbour porpoises were between 1–8 years of age. About half of them were within the age range of 4 (26%) and 5 (21%) years old, and 78% were physically immature individuals. Turbot fishing carried out with bottom nets, especially in May and June, when turbot fishing is banned, is a threat to the sustainability of harbour porpoise stocks.
Arbitral Praeliminaria – Reflections on the Distinction between Admissibility and Jurisdiction after BG v. Argentina
Authors: ROSENFELD FRIEDRICH
Published: February 2016

Arbitral tribunals have traditionally encountered difficulties in drawing the distinction between jurisdiction and admissibility. The various approaches range from the rejection of the concept of admissibility in arbitral proceedings to an overly expansive interpretation of the concept of admissibility so as to include aspects of jurisdiction. With BG Group v. Republic of Argentina, the US Supreme Court has further complicated the problem in what has become the first decision in its history on the interpretation of a bilateral investment treaty. The present article sets forth a test for distinguishing jurisdiction from admissibility which is in line with international jurisprudence and takes due account of the normative and institutional particularities of international investment arbitration proceedings.
A longitudinal study of risk factors for shedding of VTEC O157 by young cattle in herds with knownE. coliO157 carriage
Authors: SMITH R. P. , POLLITT W. J. , PAIBA G. A.
Published: February 2016

A longitudinal study in England and Wales of two dairy, five beef-fattener and three beef-suckler herds was carried out to identify risk factors for young cattle excreting verocytotoxin-producingEscherichia coliO157 (VTEC O157). A total of 1383 cattle, selected into cohorts at 0–24 months were sampled between March 2000 and February 2001. Mixed-effects logistic regression was employed to identify significant associations between VTEC O157 isolation from rectal faecal samples and explanatory factors (P< 0·001 unless shown). The results revealed a positive association with feeding root crops and a negative association with animals fed silage, milk (P= 0·001) or grain (P= 0·027). Cattle in suckler herds (P= 0·001) and those changing group between sampling visits were identified as negatively associated with VTEC O157 presence. The recovery of VTEC O157 varied throughout the year. However, the winter period from December to February was a risk factor in the multivariable analysis. Cattle in pens were 4·7 times more likely to shed VTEC O157 than those group-housed or at pasture. VTEC O157 detected in pooled environmental faecal pats and biofilm of the water supply within a group's enclosure were positively associated with an animal's VTEC O157 status in the multivariable logistic regression, as was detection of VTEC O157 in the pooled faecal pats at the previous visit.
Foot-fall and Hoof-hit. Agencies, Movements, Materialities, and Identities; and Later Prehistoric and Romano-British Trackways
Authors: Chadwick Adrian M.
Published: February 2016

In archaeological considerations of Iron Age and Romano-British landscapes, trackways are usually interpreted in purely normative terms, merely as means of getting from one settlement to another, or as functional features to assist with the herding of animals. In these somewhat static expositions, the role of trackways as places in themselves, and their long-term importance in constructions of social identity and memory, are often overlooked, as are the complex relationships between people and animals within the landscape. Recent theoretical ideas concerning relational agency and identity, materiality and movement have much to offer in terms of our archaeological understandings of these features. This paper explores the interpretative potential of such approaches using case studies of Iron Age and Romano-British trackways from Yorkshire and Nottinghamshire. Integrating theories of identity, embodiment, materiality, relationality and practice highlights the sedentarism of previous explanations, and allows for much more nuanced accounts of highly dynamic, mobile meshworks, where agency resided in complex constraints and affordances between people, animals and the materiality of the lived-in landscape.
Evidence of an advantage in visuo-spatial memory for bilingual compared to monolingual speakers
Authors: KERRIGAN LUCY , THOMAS MICHAEL S. C. , BRIGHT PETER , FILIPPI ROBERTO
Published: February 2016

Previous research has indicated that bilinguals outperform monolinguals in cognitive tasks involving spatial working memory. The present study examines evidence for this claim using a different and arguably more ecologically valid method (the change blindness task). Bilingual and monolingual participants were presented with two versions of the same scenes and required to press a key as soon as they identified the alteration. They also completed the word and alpha span tasks, and the Corsi blocks task. The results in the change blindness task, controlled for group differences in non-verbal reasoning, indicated that bilinguals were faster and more accurate than monolinguals at detecting visual changes. Similar group differences were found on the Corsi block task. Unlike previous findings, no group differences were found on the verbal memory tasks. The results are discussed with reference to mechanisms of cognitive control as a locus of transfer between bilingualism and spatial working memory tasks.
Kant’s Justification of Parental Duties
Authors: Puls Heiko
Published: February 2016

In his applied moral philosophy, Kant formulates the parents’ duty to make their child happy. I argue that, for Kant, this duty is anad hocattempt at compensating for the parental guilt of having brought a person into the condition of existence – and hence also having created her need for happiness – on their own initiative. I argue that Kant’s considerations regarding parental duties and human reproduction in general imply arguments for an ethically justified anti-natalism, but that this position is abolished in his teleology for meta-ethical reasons.
Revisiting Procedure and Precedent in the WTO: An Analysis ofUS – Countervailing and Anti-Dumping Measures (China)
Authors: BESHKAR MOSTAFA , CHILTON ADAM S.
Published: February 2016

After not applying countervailing duty (CVD) law against non-market economies (NMEs) for two decades, the United States opened a CVD investigation against China in 2006. After extensive litigation, a US appeals court ruled that it was illegal to apply CVD law to NMEs. While that ruling was being appealed, the US Congress passed legislation stipulating that the application of CVD law to NMEs starting in 2006 was legal. China challenged this legislation at the WTO. The dispute resulted in a ruling that left open the possibility that the legislation violated the GATT, as well as a finding that the United States must investigate its application of countervailing and antidumping duties against China. This dispute has implications for a number of current WTO debates including: whether Appellate Body rulings create a binding precedent, whether the Appellate Body should have authority to remand cases, and what information should be required in panel requests.
Competitive ability of rotational crops with weeds in dryland organic wheat production systems
Authors: Tautges Nicole E. , Burke Ian C. , Borrelli Kristy , Fuerst E. Patrick
Published: February 2016

While demand continues to grow and prices for organic grains have remained almost double those of conventional grains, few growers in the dryland wheat production region of Eastern Washington produce organic grain. Growers have cited weed control constraints as a top factor preventing adoption of organic production practices. In organic systems, inherent competitive ability of crops is very important in managing and preventing weed infestations. The objective of this study was to identify crop species that could reduce weed pressure and compete with perennial weeds in a wheat (Triticum aestivum) rotation. To assess weed suppressive ability of alternative rotational crops, relative total weed biomass and relative biomass of two perennial and three annual weed species were examined over 4 years in three organic cropping systems where winter wheat was in rotation with alternative crops, as part of a long-term study examining dryland organic wheat production. Three years of continuous alfalfa (Medicago sativa) production reduced Canada thistle (Cirsium arvense) biomass during the alfalfa production phase, and reduced Canada thistle biomass in winter wheat following alfalfa, compared with wheat following winter pea (Pisum sativum). Alfalfa was competitive with wild oat (Avena fatua), though it competed poorly with winter annual grass weeds. Spring barley (Hordeum vulgare), when established successfully, suppressed jointed goatgrass (Aegilops cylindrica) more than winter triticale (xTriticosecale), winter wheat, and spring wheat, and demonstrated some competitiveness toward field bindweed (Convolvulus arvensis). Winter triticale competed poorly with field bindweed, but suppressed Canada thistle and downy brome (Bromus tectorum). All alternative rotational crops contained lower weed biomass than winter pea, the crop typically rotated with wheat in the region. Organic and conventional growers could gain benefits in perennial and winter annual weed suppression by incorporating alternative rotational crops into wheat rotations.
NGF sensitizes TrkA SH-SY5Y neuroblastoma cells to TRAIL-induced apoptosis
Authors: Ruggeri P , Cappabianca L , Farina A R , Gneo L , Mackay A R
Published: February 2016

We report a novel pro-apoptotic function for nerve growth factor (NGF) and its tropomyosin-related kinase A (TrkA) receptor in sensitizing TRAIL (TNF-related apoptotis-inducing ligand)-resistant SH-SY5Y neuroblastoma (NB) cells to TRAIL-induced apoptosis, resulting in the abrogation of anchorage-independent tumourigenic growth in vitro. We show that the TRAIL-resistant SH-SY5Y phenotype is cFLIP (cellular FLICE-like inhibitory protein) dependent and not due to low-level functional TRAIL receptor or caspase expression or an inhibitory equilibrium between functional and decoy TRAIL receptors or B-cell lymphoma 2 (Bcl-2) and BH3-only (Bcl-2 homology domain 3-only) family proteins. NGF sensitization of SH-SY5Y cells to TRAIL-induced apoptosis was dependent upon TrkA expression, activation and subsequent sequestration of cFLIP. This reduces cFLIP recruitment to TRAIL-activated death receptors and increases the recruitment of caspase-8, leading to TRAIL-induced, caspase-dependent, type II apoptosis via the intrinsic mitochondrial pathway. This effect was temporary, inhibited within 6 h by nuclear factor-κ binding (NF-κB)-mediated increase in myeloid cell leukaemia-1 (Mcl-1) expression, abrogated by transient cFLIP or B-cell lymphoma-extra large (Bcl-xL) overexpression and optimized by NF-κB and Mcl-1 inhibitors. This novel mechanism adds an important pro-apoptotic immunological dimension to NGF/TrkA interaction that may not only help to explain the association between TrkA expression, better prognosis and spontaneous remission in NB, but also provides a novel potential pro-apoptotic therapeutic use for NGF, TRAIL and inhibitors of NF-κB and/or Mcl-1 in favourable and unfavourable NBs that express TrkA and exhibit cFLIP-mediated TRAIL resistance.
miR‐34a inhibits the apoptosis of MDSCs by suppressing the expression of N‐myc
Authors: Chen Si , Huang Anfei , Chen Huo , Yang Yi , Xia Fei , Jin Liping , Zhang Jinping
Published: February 2016

Myeloid‐derived suppressor cells (MDSCs) are a heterogeneous population of early myeloid progenitors, which possess strong immunosuppressive functions. MDSCs are found in increased numbers in infectious and inflammatory pathological conditions. However, whether microRNAs have a role in the expansion remains unclear. Here in our study, we found that overexpression of miR‐34a could induce the expansion of MDSCs in the bone marrow and spleen both in chimera and transgenic mice. And further experiments demonstrated that miR‐34a inhibited the apoptosis through reduced translation of N‐myc without affecting the proliferation. Luciferase assay and western blotting experiments implied that N‐myc is the direct target of miR‐34a in MDSCs. Overexpressed mir‐34a changes the cytokine expression profile in MDSCs and skewed the MDSCs to M1 phenotype. And miR‐34a‐overexpressed MDSCs significantly slowed down the tumor growth. Taken together, miR‐34a contributes to the expansion of MDSCs by inhibiting the apoptosis via suppressing the expression of N‐myc.
Correction: Corrigendum: Optically initialized robust valley-polarized holes in monolayer WSe2
Authors: Hsu Wei-Ting , Chen Yen-Lun , Chen Chang-Hsiao , Liu Pang-Shiuan , Hou Tuo-Hung , Li Lain-Jong , Chang Wen-Hao
Published: February 2016

Nature Communications 6: Article number: 8963 (2015); Published: 25 November 2015; Updated: 1 February 2016 The original version of this Article contained a typographical error in the spelling of the author Chang-Hsiao Chen, which was incorrectly given as Chiang-Hsiao Chen. This has now been corrected in both the PDF and HTML versions of the Article.
MDM2 E3 ligase-mediated ubiquitination and degradation of HDAC1 in vascular calcification
Authors: Kwon Duk-Hwa , Eom Gwang Hyeon , Ko Jeong Hyeon , Shin Sera , Joung Hosouk , Choe Nakwon , Nam Yoon Seok , Min Hyun-Ki , Kook Taewon , Yoon Somy , Kang Wanseok , Kim Yong Sook , Kim Hyung Seok , Choi Hyuck , Koh Jeong-Tae , Kim Nacksung , Ahn Youngkeun , Cho Hyun-Jai , Lee In-Kyu , Park Dong Ho , Suk Kyoungho , Seo Sang Beom , Wissing Erin R. , Mendrysa Susan M. , Nam Kwang-Il , Kook Hyun
Published: February 2016

Vascular calcification (VC) is often associated with cardiovascular and metabolic diseases. However, the molecular mechanisms linking VC to these diseases have yet to be elucidated. Here we report that MDM2-induced ubiquitination of histone deacetylase 1 (HDAC1) mediates VC. Loss of HDAC1 activity via either chemical inhibitor or genetic ablation enhances VC. HDAC1 protein, but not mRNA, is reduced in cell and animal calcification models and in human calcified coronary artery. Under calcification-inducing conditions, proteasomal degradation of HDAC1 precedes VC and it is mediated by MDM2 E3 ubiquitin ligase that initiates HDAC1 K74 ubiquitination. Overexpression of MDM2 enhances VC, whereas loss of MDM2 blunts it. Decoy peptide spanning HDAC1 K74 and RG 7112, an MDM2 inhibitor, prevent VC in vivo and in vitro. These results uncover a previously unappreciated ubiquitination pathway and suggest MDM2-mediated HDAC1 ubiquitination as a new therapeutic target in VC.
Phylogenomic and biogeographic reconstruction of the Trichinella complex
Authors: Korhonen Pasi K. , Pozio Edoardo , La Rosa Giuseppe , Chang Bill C. H. , Koehler Anson V. , Hoberg Eric P. , Boag Peter R. , Tan Patrick , Jex Aaron R. , Hofmann Andreas , Sternberg Paul W. , Young Neil D. , Gasser Robin B.
Published: February 2016

Trichinellosis is a globally important food-borne parasitic disease of humans caused by roundworms of the Trichinella complex. Extensive biological diversity is reflected in substantial ecological and genetic variability within and among Trichinella taxa, and major controversy surrounds the systematics of this complex. Here we report the sequencing and assembly of 16 draft genomes representing all 12 recognized Trichinella species and genotypes, define protein-coding gene sets and assess genetic differences among these taxa. Using thousands of shared single-copy orthologous gene sequences, we fully reconstruct, for the first time, a phylogeny and biogeography for the Trichinella complex, and show that encapsulated and non-encapsulated Trichinella taxa diverged from their most recent common ancestor ∼21 million years ago (mya), with taxon diversifications commencing ∼10−7 mya.
The limits of precision monomer placement in chain growth polymerization
Authors: Gody Guillaume , Zetterlund Per B. , Perrier Sébastien , Harrisson Simon
Published: February 2016

Precise control over the location of monomers in a polymer chain has been described as the ‘Holy Grail’ of polymer synthesis. Controlled chain growth polymerization techniques have brought this goal closer, allowing the preparation of multiblock copolymers with ordered sequences of functional monomers. Such structures have promising applications ranging from medicine to materials engineering. Here we show, however, that the statistical nature of chain growth polymerization places strong limits on the control that can be obtained. We demonstrate that monomer locations are distributed according to surprisingly simple laws related to the Poisson or beta distributions. The degree of control is quantified in terms of the yield of the desired structure and the standard deviation of the appropriate distribution, allowing comparison between different synthetic techniques. This analysis establishes experimental requirements for the design of polymeric chains with controlled sequence of functionalities, which balance precise control of structure with simplicity of synthesis.
Complex Greenland outlet glacier flow captured
Authors: Aschwanden Andy , Fahnestock Mark A. , Truffer Martin
Published: February 2016

The Greenland Ice Sheet is losing mass at an accelerating rate due to increased surface melt and flow acceleration in outlet glaciers. Quantifying future dynamic contributions to sea level requires accurate portrayal of outlet glaciers in ice sheet simulations, but to date poor knowledge of subglacial topography and limited model resolution have prevented reproduction of complex spatial patterns of outlet flow. Here we combine a high-resolution ice-sheet model coupled to uniformly applied models of subglacial hydrology and basal sliding, and a new subglacial topography data set to simulate the flow of the Greenland Ice Sheet. Flow patterns of many outlet glaciers are well captured, illustrating fundamental commonalities in outlet glacier flow and highlighting the importance of efforts to map subglacial topography. Success in reproducing present day flow patterns shows the potential for prognostic modelling of ice sheets without the need for spatially varying parameters with uncertain time evolution.
Engineering of frustration in colloidal artificial ices realized on microfeatured grooved lattices
Authors: Ortiz-Ambriz Antonio , Tierno Pietro
Published: February 2016

Artificial spin ice systems, namely lattices of interacting single domain ferromagnetic islands, have been used to date as microscopic models of frustration induced by lattice topology, allowing for the direct visualization of spin arrangements and textures. However, the engineering of frustrated ice states in which individual spins can be manipulated in situ and the real-time observation of their collective dynamics remain both challenging tasks. Inspired by recent theoretical advances, here we realize a colloidal version of an artificial spin ice system using interacting polarizable particles confined to lattices of bistable gravitational traps. We show quantitatively that ice-selection rules emerge in this frustrated soft matter system by tuning the strength of the pair interactions between the microscopic units. Via independent control of particle positioning and dipolar coupling, we introduce monopole-like defects and strings and use loops with defined chirality as an elementary unit to store binary information.
Two betweenness centrality measures based on Randomized Shortest Paths
Authors: Kivimäki Ilkka , Lebichot Bertrand , Saramäki Jari , Saerens Marco
Published: February 2016

This paper introduces two new closely related betweenness centrality measures based on the Randomized Shortest Paths (RSP) framework, which fill a gap between traditional network centrality measures based on shortest paths and more recent methods considering random walks or current flows. The framework defines Boltzmann probability distributions over paths of the network which focus on the shortest paths, but also take into account longer paths depending on an inverse temperature parameter. RSP’s have previously proven to be useful in defining distance measures on networks. In this work we study their utility in quantifying the importance of the nodes of a network. The proposed RSP betweenness centralities combine, in an optimal way, the ideas of using the shortest and purely random paths for analysing the roles of network nodes, avoiding issues involving these two paradigms. We present the derivations of these measures and how they can be computed in an efficient way. In addition, we show with real world examples the potential of the RSP betweenness centralities in identifying interesting nodes of a network that more traditional methods might fail to notice.
Uncertainty principle for experimental measurements: Fast versus slow probes
Authors: Hansmann P. , Ayral T. , Tejeda A. , Biermann S.
Published: February 2016

The result of a physical measurement depends on the time scale of the experimental probe. In solid-state systems, this simple quantum mechanical principle has far-reaching consequences: the interplay of several degrees of freedom close to charge, spin or orbital instabilities combined with the disparity of the time scales associated to their fluctuations can lead to seemingly contradictory experimental findings. A particularly striking example is provided by systems of adatoms adsorbed on semiconductor surfaces where different experiments – angle-resolved photoemission, scanning tunneling microscopy and core-level spectroscopy – suggest different ordering phenomena. Using most recent first principles many-body techniques, we resolve this puzzle by invoking the time scales of fluctuations when approaching the different instabilities. These findings suggest a re-interpretation of ordering phenomena and their fluctuations in a wide class of solid-state systems ranging from organic materials to high-temperature superconducting cuprates.
Universal conformational properties of polymers in ionic nanogels
Authors: Kobayashi Hideki , Winkler Roland G.
Published: February 2016

Polyelectrolyte gels are known to undergo significant conformational changes in response to external stimuli such as pH, temperature, or the dielectric constant. Specifically, an increase of the degree of ionization associated with an increasing number of counterions leads to swelling of the network. For a macroscopically large gel, which is electrostatically neutral in its interior, swelling is no longer governed by electrostatic interactions, but rather by the osmotic pressure of counterions. However, this electrostatic neutrality is typically violated for nanogels, because counterions are free to leave a gel particle. Although nanogel-swelling exhibits similar features as swelling of micro- and macrogels, another mechanism has to be relevant. Here, we use molecular dynamics simulations and scaling theory to unravel the structural properties of nanogels upon changing the electrostatic interactions. We demonstrate that the swelling of nanogels is governed by screened electrostatic interactions without a relevant contribution by the counterion osmotic pressure.
Short-term memory trace mediated by termination kinetics of olfactory receptor
Authors: Boyle Sean Michael , McInally Shane , Tharadra Sana , Ray Anandasankar
Published: February 2016

Odorants activate receptors in the peripheral olfactory neurons, which sends information to higher brain centers where behavioral valence is determined. Movement and airflow continuously change what odor plumes an animal encounters and little is known about the effect one plume has on the detection of another. Using the simple Drosophila melanogaster larval model to study this relationship we identify an unexpected phenomenon: response to an attractant can be selectively blocked by previous exposure to some odorants that activates the same receptor. At a mechanistic level, we find that exposure to this type of odorant causes prolonged tonic responses from a receptor (Or42b), which can block subsequent detection of a strong activator of that same receptor. We identify naturally occurring odorants with prolonged tonic responses for other odorant receptors (Ors) as well, suggesting that termination-kinetics is a factor for olfactory coding mechanisms. This mechanism has implications for odor-coding in any system and for designing applications to modify odor-driven behaviors.
ClC-7 Deficiency Impairs Tooth Development and Eruption
Authors: Wang He , Pan Meng , Ni Jinwen , Zhang Yanli , Zhang Yutao , Gao Shan , Liu Jin , Wang Zhe , Zhang Rong , He Huiming , Wu Buling , Duan Xiaohong
Published: February 2016

CLCN7 gene encodes the voltage gated chloride channel 7 (ClC-7) in humans. The mutations in CLCN7 have been associated with osteopetrosis in connection to the abnormal osteoclasts functions. Previously, we found that some osteopetrosis patients with CLCN7 mutations suffered from impacted teeth and root dysplasia. Here we set up two in vivo models under a normal or an osteoclast-poor environment to investigate how ClC-7 affects tooth development and tooth eruption. Firstly, chitosan-Clcn7-siRNA nanoparticles were injected around the first maxillary molar germ of newborn mice and caused the delay of tooth eruption and deformed tooth with root dysplasia. Secondly, E13.5 molar germs infected with Clcn7 shRNA lentivirus were transplanted under the kidney capsule and presented the abnormal changes in dentin structure, periodontal tissue and cementum. All these teeth changes have been reported in the patients with CLCN7 mutation. In vitro studies of ameloblasts, odontoblasts and dental follicle cells (DFCs) were conducted to explore the involved mechanism. We found that Clcn7 deficiency affect the differentiation of these cells, as well as the interaction between DFCs and osteoclasts through RANKL/OPG pathway. We conclude that ClC-7 may affect tooth development by directly targeting tooth cells and regulate tooth eruption through DFC mediated osteoclast pathway.
Resveratrol analogue 4,4′-dihydroxy-trans-stilbene potently inhibits cancer invasion and metastasis
Authors: Savio Monica , Ferraro Daniela , Maccario Cristina , Vaccarone Rita , Jensen Lasse D. , Corana Federica , Mannucci Barbara , Bianchi Livia , Cao Yihai , Stivala Lucia Anna
Published: February 2016

We investigated the preventive effects of resveratrol analogue 4,4′-dihydroxy-trans-stilbene (DHS) on cancer invasion and metastasis. Two different in vivo approaches of mouse and zebrafish lung cancer invasion models were employed in our study. The in vitro results showed that DHS displays potent inhibition on anchorage-dependent or -independent cell growth of LLC cells, leading to impairment of the cell cycle progression with reduction of cell numbers arresting at the G1 phase, an evident accumulation of pre-G1 events correlated with apoptotic behaviour. In addition, DHS induces a marked inhibition of LLC cell migration and matrigel invasion. In a murine lung cancer model, tumour volume, cell proliferation and tumour angiogenesis were significantly inhibited by DHS. Importantly, liver metastatic lesions were significantly reduced in DHS-treated mice. Similarly, DHS significantly inhibits lung cancer cell dissemination, invasion and metastasis in a zebrafish tumour model. These findings demonstrate that DHS could potentially be developed as a novel therapeutic agent for treatment of cancer and metastasis.
Broad and potent antiviral activity of the NAE inhibitor MLN4924
Authors: Le-Trilling Vu Thuy Khanh , Megger Dominik A. , Katschinski Benjamin , Landsberg Christine D. , Rückborn Meike U. , Tao Sha , Krawczyk Adalbert , Bayer Wibke , Drexler Ingo , Tenbusch Matthias , Sitek Barbara , Trilling Mirko
Published: February 2016

In terms of infected human individuals, herpesviruses range among the most successful virus families. Subclinical herpesviral infections in healthy individuals contrast with life-threatening syndromes under immunocompromising and immunoimmature conditions. Based on our finding that cytomegaloviruses interact with Cullin Roc ubiquitin ligases (CRLs) in the context of interferon antagonism, we systematically assessed viral dependency on CRLs by utilizing the drug MLN4924. CRL activity is regulated through the conjugation of Cullins with the ubiquitin-like molecule Nedd8. By inhibiting the Nedd8-activating Enzyme (NAE), MLN4924 interferes with Nedd8 conjugation and CRL activity. MLN4924 exhibited pronounced antiviral activity against mouse and human cytomegalovirus, herpes simplex virus (HSV)- 1 (including multi-drug resistant clinical isolates), HSV-2, adeno and influenza viruses. Human cytomegalovirus genome amplification was blocked at nanomolar MLN4924 concentrations. Global proteome analyses revealed that MLN4924 blocks cytomegaloviral replication despite increased IE1 amounts. Expression of dominant negative Cullins assigned this IE regulation to defined Cullin molecules and phenocopied the antiviral effect of MLN4924.
Toward Contactless Biology: Acoustophoretic DNA Transfection
Authors: Vasileiou Thomas , Foresti Daniele , Bayram Adem , Poulikakos Dimos , Ferrari Aldo
Published: February 2016

Acoustophoresis revolutionized the field of container-less manipulation of liquids and solids by enabling mixing procedures which avoid contamination and loss of reagents due to the contact with the support. While its applications to chemistry and engineering are straightforward, additional developments are needed to obtain reliable biological protocols in a contactless environment. Here, we provide a first, fundamental step towards biological reactions in air by demonstrating the acoustophoretic DNA transfection of mammalian cells. We developed an original acoustophoretic design capable of levitating, moving and mixing biological suspensions of living mammalians cells and of DNA plasmids. The precise and sequential delivery of the mixed solutions into tissue culture plates is actuated by a novel mechanism based on the controlled actuation of the acoustophoretic force. The viability of the contactless procedure is tested using a cellular model sensitive to small perturbation of neuronal differentiation pathways. Additionally, the efficiency of the transfection procedure is compared to standard, container-based methods for both single and double DNA transfection and for different cell types including adherent growing HeLa cancer cells and low adhesion neuron-like PC12 cells. In all, this work provides a proof of principle which paves the way to the development of high-throughput acoustophoretic biological reactors.
Increased serum microRNAs are closely associated with the presence of microvascular complications in type 2 diabetes mellitus
Authors: Wang Cheng , Wan Shujun , Yang Ting , Niu Dongmei , Zhang Aisen , Yang Cuihua , Cai Jialu , Wu Jia , Song Jiaxi , Zhang Chen-Yu , Zhang Chunni , Wang Junjun
Published: February 2016

Circulating microRNAs (miRNAs) are emerging biomarkers for type 2 diabetes mellitus (T2DM). However, a comprehensive characterization of the serum miRNA profile in patients with T2DM-associated microvascular disease (T2DMC) has rarely been reported. In this study, we obtained serum samples from 184 T2DM patients (92 with microvascular complications and 92 free of complications) and 92 age/gender-matched controls. The levels of 754 miRNAs were initially analyzed using a TaqMan Low Density Array (TLDA) in three pooled samples from 24 T2DM patients, 24 T2DMC patients and 24 controls. Markedly upregulated miRNAs in the patients’ groups were subsequently validated individually by quantitative reverse-transcription PCR (RT-qPCR) in the same samples used for TLDA and further confirmed in another larger cohort consisting of 68 patients with T2DM, 68 patients with T2DMC and 68 controls. Five miRNAs were significantly upregulated in T2DM patients (p < 0.05) including miR-661, miR-571, miR-770-5p, miR-892b and miR-1303. Moreover, the levels of the five miRNAs were higher in patients with complications than in those without complications. Regression analyses revealed the five miRNAs were significantly correlated with microvascular complications (p < 0.05). The five serum miRNAs identified in our study hold potential as auxiliary biomarkers and novel risk factors for T2DM-associated microvascular complications.
Gene profiling of embryonic skeletal muscle lacking type I ryanodine receptor Ca2+ release channel
Authors: Filipova Dilyana , Walter Anna M. , Gaspar John A. , Brunn Anna , Linde Nina F. , Ardestani Mostafa A. , Deckert Martina , Hescheler Jürgen , Pfitzer Gabriele , Sachinidis Agapios , Papadopoulos Symeon
Published: February 2016

In mature skeletal muscle, the intracellular Ca2+ concentration rises dramatically upon membrane depolarization, constituting the link between excitation and contraction. This process requires Ca2+ release from the sarcoplasmic reticulum via the type 1 ryanodine receptor (RYR1). However, RYR1’s potential roles in muscle development remain obscure. We used an established RyR1- null mouse model, dyspedic, to investigate the effects of the absence of a functional RYR1 and, consequently, the lack of RyR1-mediated Ca2+ signaling, during embryogenesis. Homozygous dyspedic mice die after birth and display small limbs and abnormal skeletal muscle organization. Skeletal muscles from front and hind limbs of dyspedic fetuses (day E18.5) were subjected to microarray analyses, revealing 318 differentially expressed genes. We observed altered expression of multiple transcription factors and members of key signaling pathways. Differential regulation was also observed for genes encoding contractile as well as muscle-specific structural proteins. Additional qRT-PCR analysis revealed altered mRNA levels of the canonical muscle regulatory factors Six1, Six4, Pax7, MyoD, MyoG and MRF4 in mutant muscle, which is in line with the severe developmental retardation seen in dyspedic muscle histology analyses. Taken together, these findings suggest an important non-contractile role of RyR1 or RYR1-mediated Ca2+ signaling during muscle organ development.
In vivo longitudinal study of rodent skeletal muscle atrophy using ultrasonography
Authors: Mele Antonietta , Fonzino Adriano , Rana Francesco , Camerino Giulia Maria , De Bellis Michela , Conte Elena , Giustino Arcangela , Conte Camerino Diana , Desaphy Jean-François
Published: February 2016

Muscle atrophy is a widespread ill condition occurring in many diseases, which can reduce quality of life and increase morbidity and mortality. We developed a new method using non-invasive ultrasonography to measure soleus and gastrocnemius lateralis muscle atrophy in the hindlimb-unloaded rat, a well-accepted model of muscle disuse. Soleus and gastrocnemius volumes were calculated using the conventional truncated-cone method and a newly-designed sinusoidal method. For Soleus muscle, the ultrasonographic volume determined in vivo with either method was linearly correlated to the volume determined ex-vivo from excised muscles as muscle weight-to-density ratio. For both soleus and gastrocnemius muscles, a strong linear correlation was obtained between the ultrasonographic volume and the muscle fiber cross-sectional area determined ex-vivo on muscle cryosections. Thus ultrasonography allowed the longitudinal in vivo evaluation of muscle atrophy progression during hindlimb unloading. This study validates ultrasonography as a powerful method for the evaluation of rodent muscle atrophy in vivo, which would prove useful in disease models and therapeutic trials.
Inferring the Dynamics of Effective Population Size Using Autosomal Genomes
Authors: Hou Zheng , Luo Yin , Wang Zhisheng , Zheng Hong-Xiang , Wang Yi , Zhou Hang , Wu Leqin , Jin Li
Published: February 2016

Next-generation sequencing technology has provided a great opportunity for inferring human demographic history by investigating changes in the effective population size (Ne). In this report, we introduce a strategy for estimating Ne dynamics, allowing the exploration of large multi-locus SNP datasets. We applied this strategy to the Phase 1 Han Chinese samples from the 1000 Genomes Project. The Han Chinese population has undergone a continuous expansion since 25,000 years ago, at first slowly from about 7,300 to 9,800 (at the end of the last glacial maximum about 15,000 YBP), then more quickly to about 46,000 (at the beginning of the Neolithic about 8,000 YBP) and then even more quickly to reach a population size of about 140,000 (recently).
Fifteen new earthworm mitogenomes shed new light on phylogeny within the Pheretima complex
Authors: Zhang Liangliang , Sechi Pierfrancesco , Yuan Minglong , Jiang Jibao , Dong Yan , Qiu Jiangping
Published: February 2016

The Pheretima complex within the Megascolecidae family is a major earthworm group. Recently, the systematic status of the Pheretima complex based on morphology was challenged by molecular studies. In this study, we carry out the first comparative mitogenomic study in oligochaetes. The mitogenomes of 15 earthworm species were sequenced and compared with other 9 available earthworm mitogenomes, with the main aim to explore their phylogenetic relationships and test different analytical approaches on phylogeny reconstruction. The general earthworm mitogenomic features revealed to be conservative: all genes encoded on the same strand, all the protein coding loci shared the same initiation codon (ATG) and tRNA genes showed conserved structures. The Drawida japonica mitogenome displayed the highest A + T content, reversed AT/GC-skews and the highest genetic diversity. Genetic distances among protein coding genes displayed their maximum and minimum interspecific values in the ATP8 and CO1 genes, respectively. The 22 tRNAs showed variable substitution patterns between the considered earthworm mitogenomes. The inclusion of rRNAs positively increased phylogenetic support. Furthermore, we tested different trimming tools for alignment improvement. Our analyses rejected reciprocal monophyly among Amynthas and Metaphire and indicated that the two genera should be systematically classified into one.
Genetic Variations of NLR family genes in Behcet’s Disease
Authors: Li Lin , Yu Hongsong , Jiang Yanni , Deng Bolin , Bai Lin , Kijlstra Aize , Yang Peizeng
Published: February 2016

This study aimed to investigate whether single nucleotide polymorphisms (SNPs) of five NLR family genes (NOD1, NOD2, NLRP1, NLRP3 and CIITA) are associated with Behcet’s disease (BD) in a Chinese Han population. The study was carried out in 950 BD patients and 1440 controls for 19 SNPs in the selected NLR genes. In the first-stage study, significantly decreased frequencies of the CIITA//rs12932187 C allele (Pc = 1.668E-02) and NOD1//rs2075818 G allele (Pc = 4.694E-02) were found in BD patients as compared to controls . After performing a second stage validation study and combination of data we confirmed the association of CIITA//rs12932187 and NOD1//rs2075818 with BD. In CIITA//rs12932187, the frequencies of the CC genotype and C allele were significantly lower in BD than in controls (Pc = 3.331E-06; Pc = 6.004E-07, respectively). In NOD1//rs2075818, the GG genotype and G allele showed significantly decreased frequencies in BD patients when compared to controls (Pc = 1.022E-02; Pc = 6.811E-05, respectively). Functional experiments showed that carriers with the CC genotype in CIITA//rs12932187 had a lower CIITA mRNA expression level and an enhanced IL-10 secretion as compared to GG and CG carriers. This study provides evidence that the CIITA and NOD1 gene are involved in the susceptibility to Behcet’s disease.
Dusky-like is required for epidermal pigmentation and metamorphosis in Tribolium castaneum
Authors: Li Chengjun , Yun Xiaopei , Li Bin
Published: February 2016

Dusky-like (Dyl) is associated with the morphogenesis of embryonic denticle, adult sensory bristle and wing hair in Drosophila melanogaster. And whether Dyl involved in insect post-embryonic development and its signal transduction are poorly understood. Here, phylogenetic analysis revealed that dyl displayed one-to-one orthologous relationship among insects. In Tribolium castaneum, dyl is abundantly expressed at the late embryonic stage. Tissue-specific expression analysis at the late adult stage illustrated high expression of dyl in the fat body and ovary. Knockdown of dyl resulted in the defects in larval epidermal pigmentation and completely blocked the transitions from larval to pupal and pupal to adult stages of T. castaneum. We further discovered that dyl RNAi phenotypes were phenocopied by blimp-1 or shavenbaby (svb) silencing and dyl was positively regulated by blimp-1 through svb in T. castaneum. These results suggest that Dyl functions downstream of Blimp-1 through Svb for larval epidermal pigmentation and metamorphosis. Moreover, ftz-f1 was down-regulated after RNA interference (RNAi) suppressing any of those three genes, indicating that Ftz-f1 works downstream of Dyl to mediate the effects of Blimp-1, Svb and Dyl on metamorphosis in T. castaneum. This study provides valuable insights into functions and signaling pathway of insect Dyl.
Phenanthrene exposure induces cardiac hypertrophy via reducing miR-133a expression by DNA methylation
Authors: Huang Lixing , Xi Zhihui , Wang Chonggang , Zhang Youyu , Yang Zhibing , Zhang Shiqi , Chen Yixin , Zuo Zhenghong
Published: February 2016

Growing evidence indicates that there is an emerging link between environmental pollution and cardiac hypertrophy, while the mechanism is unclear. The objective of this study was to examine whether phenanthrene (Phe) could cause cardiac hypertrophy and elucidate the molecular mechanisms involved. We found that: 1) Phe exposure increased the heart weight and cardiomyocyte size of rats; 2) Phe exposure led to enlarged cell size and increased protein synthesis in H9C2 cells; 3) Phe exposure induced important markers of cardiac hypertrophy, such as atrial natriuretic peptide, B-type natriuretic peptide and c-Myc in H9C2 cells and rat hearts; 4) Phe exposure perturbed miR-133a, CdC42 and RhoA, which were key regulators of cardiac hypertrophy, in H9C2 cells and rat hearts; 5) Phe exposure induced DNA methyltransferases (DNMTs) in H9C2 cells and rat hearts; 6) Phe exposure led to methylation of CpG sites within the miR-133a locus and reduced miR-133a expression in H9C2 cells; 7) DNMT inhibition and miR-133a overexpression could both alleviate the enlargement of cell size and perturbation of CdC42 and RhoA caused by Phe exposure. These results indicated that Phe could induce cardiomyocyte hypertrophy in the rat and H9C2 cells. The mechanism might involve reducing miR-133a expression by DNA methylation.
LIPUS promotes spinal fusion coupling proliferation of type H microvessels in bone
Authors: Xu Ximing , Wang Fei , Yang Yahong , Zhou Xiaoyi , Cheng Yajun , Wei Xianzhao , Li Ming
Published: February 2016

Low-intensity pulsed ultrasound (LIPUS) has been found to accelerate spinal fusion. Type H microvessels are found in close relation with bone development. We analyzed the role of type H vessels in rat spinal fusion model intervened by LIPUS. It was found LIPUS could significantly accelerate bone fusion rate and enlarge bone callus. Osteoblasts were specifically located on the bone meshwork of the allograft and were surrounded by type H microvessels. LIPUS could significantly increase the quantity of osteoblasts during spine fusion, which process was coupled with elevated angiogenesis of type H microvessels. Our results suggest that LIPUS may be a noninvasive adjuvant treatment modality in spinal fusion for clinical use. The treatment is recommended for usage for at least one month.