bluuebunny/crossref_metadata_2025 huggingface dataset:
Linking Microbial Community and Catabolic Gene Structures during the Adaptation of Three Contaminated Soils under Continuous Long-Term Pollutant Stress
Authors: Lima-Morales Daiana , Jáuregui Ruy , Camarinha-Silva Amelia , Geffers Robert , Pieper Dietmar H. , Vilchez-Vargas Ramiro
Published: February 2016

Three types of contaminated soil from three geographically different areas were subjected to a constant supply of benzene or benzene/toluene/ethylbenzene/xylenes (BTEX) for a period of 3 months. Different from the soil from Brazil (BRA) and Switzerland (SUI), the Czech Republic (CZE) soil which was previously subjected to intensive
in situ
bioremediation displayed only negligible changes in community structure. BRA and SUI soil samples showed a clear succession of phylotypes. A rapid response to benzene stress was observed, whereas the response to BTEX pollution was significantly slower. After extended incubation, actinobacterial phylotypes increased in relative abundance, indicating their superior fitness to pollution stress. Commonalities but also differences in the phylotypes were observed. Catabolic gene surveys confirmed the enrichment of actinobacteria by identifying the increase of actinobacterial genes involved in the degradation of pollutants. Proteobacterial phylotypes increased in relative abundance in SUI microcosms after short-term stress with benzene, and catabolic gene surveys indicated enriched metabolic routes. Interestingly, CZE soil, despite staying constant in community structure, showed a change in the catabolic gene structure. This indicates that a highly adapted community, which had to adjust its gene pool to meet novel challenges, has been enriched.
New Recombinant Mycobacterium bovis BCG Expression Vectors: Improving Genetic Control over Mycobacterial Promoters
Authors: Kanno Alex I. , Goulart Cibelly , Rofatto Henrique K. , Oliveira Sergio C. , Leite Luciana C. C. , McFadden Johnjoe
Published: February 2016

The expression of many antigens, stimulatory molecules, or even metabolic pathways in mycobacteria such as
Mycobacterium bovis
BCG or
M. smegmatis
was made possible through the development of shuttle vectors, and several recombinant vaccines have been constructed. However, gene expression in any of these systems relied mostly on the selection of natural promoters expected to provide the required level of expression by trial and error. To establish a systematic selection of promoters with a range of strengths, we generated a library of mutagenized promoters through error-prone PCR of the strong P
L5
promoter, originally from mycobacteriophage L5. These promoters were cloned upstream of the enhanced green fluorescent protein reporter gene, and recombinant
M. smegmatis
bacteria exhibiting a wide range of fluorescence levels were identified. A set of promoters was selected and identified as having high (pJK-F8), intermediate (pJK-B7, pJK-E6, pJK-D6), or low (pJK-C1) promoter strengths in both
M. smegmatis
and
M. bovis
BCG. The sequencing of the promoter region demonstrated that it was extensively modified (6 to 11%) in all of the plasmids selected. To test the functionality of the system, two different expression vectors were demonstrated to allow corresponding expression levels of the
Schistosoma mansoni
antigen Sm29 in BCG. The approach used here can be used to adjust expression levels for synthetic and/or systems biology studies or for vaccine development to maximize the immune response.
Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies
Authors: Scharinger Eva J. , Dietrich Richard , Kleinsteuber Ina , Märtlbauer Erwin , Schauer Kristina
Published: February 2016

Cronobacter sakazakii
is a foodborne pathogen associated with rare but often lethal infections in neonates. Powdered infant formula (PIF) represents the most frequent source of infection. Out of the identified serotypes (O1 to O7), O1, O2, and O3 are often isolated from clinical and PIF samples. Serotype-specific monoclonal antibodies (MAbs) suitable for application in enzyme immunoassays (EIAs) for the rapid detection of
C. sakazakii
have not yet been developed. In this study, we created specific MAbs with the ability to bind to
C. sakazakii
of serotypes O1, O2, and O3. Characterization by indirect EIAs, immunofluorescence, motility assays, and immunoblotting identified lipopolysaccharide (LPS) and exopolysaccharide (EPS) as the antigenic determinants of the MAbs. The established sandwich EIAs were highly sensitive and were able to detect between 2 × 10
3
and 9 × 10
6
CFU/ml. Inclusivity tests confirmed that 93% of serotype O1 strains, 100% of O2 strains, and 87% of O3 strains were detected at low cell counts. No cross-reactivity with >100 strains of
Cronobacter
spp. and other
Enterobacteriaceae
was observed, except for that with
C. sakazakii
serotype O3 and
Cronobacter
muytjensii
serotype O1. Moreover, the sandwich EIAs detected
C. sakazakii
in PIF samples artificially contaminated with 1 to 10 bacterial cells per 10 g of sample after 15 h of preenrichment. The use of these serotype-specific MAbs not only allows the reliable detection of
C. sakazakii
strains but also enables simultaneous serotyping in a simple sandwich EIA method.
Ileal pouch functional outlet obstruction
Authors: Sierakowski Kyra , Rabbitt Philippa , Wattchow David
Published: February 2016

Small bowel obstructions (SBOs) are common in patients who have undergone Ileal J-pouch-anal anastomosis (IPAA) surgery. SBO may be caused by stenosis of the diverting ileostomy, volvulus, internal hernia, adhesive bands, anastomotic stricture1or intra-abdominal adhesions.2Functional outlet obstruction is an important alternative diagnosis to consider in a patient post-IPAA presenting with obstructive symptoms. Recognition of this condition can prevent unnecessary surgery and save the patient from presenting repeatedly with obstructive symptoms.
Complications of indwelling pleural catheter use and their management
Authors: Lui Macy M S , Thomas Rajesh , Lee Y C Gary
Published: February 2016

The growing utilisation of indwelling pleural catheters (IPCs) has put forward a new era in the management of recurrent symptomatic pleural effusions. IPC use is safe compared to talc pleurodesis, though complications can occur. Pleural infection affects <5% of patients, and is usually responsive to antibiotic treatment without requiring catheter removal or surgery. Pleural loculations develop over time, limiting drainage in 10% of patients, which can be improved with intrapleural fibrinolytic therapy. Catheter tract metastasis can occur with most tumours but is more common in mesothelioma. The metastases usually respond to analgaesics and/or external radiotherapy. Long-term intermittent drainage of exudative effusions or chylothorax can potentially lead to loss of nutrients, though no data exist on any clinical impact. Fibrin clots within the catheter lumen can result in blockage. Chest pain following IPC insertion is often mild, and adjustments in analgaesics and drainage practice are usually all that are required. As clinical experience with the use of IPC accumulates, the profile and natural course of complications are increasingly described. We aim to summarise the available literature on IPC-related complications and the evidence to support specific strategies.
Technical and clinical accuracy of three blood glucose meters: clinical impact assessment using error grid analysis and insulin sliding scales
Authors: Sutheran Hannah Lise , Reynolds Timothy
Published: February 2016

Aims

Manufacturers of point-of-care testing (POCT) glucose systems must adhere to International Organisation for Standardisation (ISO) 15197 performance standards, a tightened version of which becomes mandatory in May 2016. Using methods deviating from the evaluation requirements of ISO 15197, we investigated the accuracy and clinical utility of three glucose testing systems.

Methods

Whole blood glucose was measured on 62 EDTA-preserved samples with each system followed by plasma glucose determination with a laboratory hexokinase method. Technical accuracy was assessed using linear regression, Bland-Altman plots and bias. Clinical utility was evaluated using consensus error grid analysis and insulin dosing algorithms.

Results

The correlation coefficient of the glucose measuring devices with the comparator method was between 0.979 and 0.993 (n=61). Bias against the reference method was +0.7–+4.4%. The FreeStyle Lite (0.7±4.4%) and ACCU-CHEK Aviva (1.3±7.3%) systems had the least bias to the reference. Error grid analysis depicts all meters to be clinically accurate, with all results falling within Zones A and B.

Conclusions

Conformité Européene-marked devices are presumed to have adequate accuracy, but postmarket evaluations are imperative to assess total system accuracy and whether regulatory targets are achieved.
Republished: Bilateral and symmetric isolated dorsospinal arteries with prominent anterior radiculomedullary contribution
Authors: Orru’ Emanuele , Gailloud Philippe
Published: February 2016

We present the case of a 15-year-old girl with a left T7 spinal epidural arteriovenous fistula (SEAVF), in whom bilateral and symmetric isolated T9 dorsospinal arteries (DAs) were incidentally documented, each providing a significant contribution to the anterior spinal artery. Recognizing the existence of isolated DAs of direct aortic origin is critical, both for the diagnostic evaluation and for the management of spinal vascular diseases. Notably, isolated DAs tend to branch off significant radiculomedullary arteries, which vascularize the normal spinal cord but have also been shown to supply spinal vascular malformations.
Continuing medical education for general practitioners: a practice format
Authors: VanNieuwenborg Lena , Goossens Martine , De Lepeleire Jan , Schoenmakers Birgitte
Published: February 2016

Introduction

Our current knowledge-based society and the many actualisations within the medical profession require a great responsibility of physicians to continuously develop and refine their skills. In this article, we reflect on some recent findings in the field of continuing education for professional doctors (continuing medical education, CME). Second, we describe the development of a CME from the Academic Center for General Practice (ACHG) of the KU Leuven.

Methods

First, we performed a literature study and we used unpublished data of a need assessment performed (2013) in a selected group of general practitioners. Second, we describe the development of a proposal to establish a CME programme for general practitioners.

Results

CME should go beyond the sheer acquisition of knowledge, and also seek changes in practice, attitudes and behaviours of physicians. The continuing education offerings are subject to the goals of the organising institution, but even more to the needs and desires of the end user.

Conclusions

Integrated education is crucial to meet the conditions for efficient and effective continuing education. The ACHG KU Leuven decided to offer a postgraduate programme consisting of a combination of teaching methods: online courses (self-study), contact courses (traditional method) and a materials database.
Cardiac manifestations in alcoholic liver disease
Authors: Milić Sandra , Lulić Davorka , Štimac Davor , Ružić Alen , Zaputović Luka
Published: February 2016

Alcoholic liver disease is the most prevalent cause of progressive liver disease in Europe. Alcoholic cirrhosis occurs in 8%–20% of cases of alcoholic liver disease. It has significant influence on cardiovascular system and haemodynamics through increased heart rate, cardiac output, decreased systemic vascular resistance, arterial pressure and plasma volume expansion. Cirrhotic cardiomyopathy is characterised by systolic and diastolic dysfunction and electrophysiological abnormalities, if no other underlying cardiac disease is present. It is often unmasked only during pharmacological or physiological stress, when compensatory mechanisms of the heart become insufficient to maintain adequate cardiac output. Low-to-moderate intake of alcohol can be cardioprotective. However, heavy drinking is associated with an increased risk of cardiovascular diseases, such as alcoholic cardiomyopathy, arterial hypertension, atrial arrhythmias as well as haemorrhagic and ischaemic stroke. Alcoholic cardiomyopathy is characterised by dilated left ventricle (LV), increased LV mass, normal or reduced LV wall thickness and systolic dysfunction.
Influence of forest composition and age on habitat use by bats in southwestern British Columbia
Authors: Luszcz Tanya M.J. , Barclay Robert M.R.
Published: February 2016

Habitat use by bats in forests varies with environmental variables, clutter, and prey availability, but few studies have looked at habitat use as a function of forest composition and forest age across a range of elevations. We examined factors influencing habitat use of forest-dwelling bats in southwestern British Columbia using ultrasonic detection. We measured activity of three bat foraging guilds in four forest types and three age classes from May to August 2000 and 2001. Habitat use varied with forest composition and stand age, but not always as we predicted. Activity of open-habitat foragers (large bats) was greatest in higher elevation forests and in young forests. In contrast, activity of the long-eared myotis (Myotis evotis (H. Allen, 1864)), a gleaner, was greater in lower elevation black cottonwood (Populus trichocarpa Torr. & A. Gray ex Hook.) and cedar–hemlock stands. Myotis evotis activity also increased with increasing forest age. Activity of edge and gap insectivores (genus Myotis Kaup, 1829) was greater in black cottonwood stands and old Interior Douglas-fir stands than in other forest types. The probability of detecting foraging Myotis was highest in black cottonwood stands. Myotis activity did not show a clear pattern with forest age. Our results demonstrate the importance of black cottonwood riparian forests to foraging Myotis, as well as the importance of older, low-elevation forests.
Fibrous gypsum veins as diffuse features and within fault zones: the case study of the Pisco Basin (Ica desert, southern Peru)
Authors: Rustichelli Andrea , Di Celma Claudio , Tondi Emanuele , Baud Patrick , Vinciguerra Sergio
Published: February 2016

New knowledge on patterns of fibrous gypsum veins, their genetic mechanisms, deformation style and weathering is provided by a field- and laboratory-based study carried out on the Neogene to Quaternary Pisco Basin sedimentary strata exposed in the Ica desert, southern Peru. Gypsum veins vary considerably in dimensions, attitudes and timing, and can develop in layered and moderately fractured rocks also in the absence of evaporitic layers. Veins occur both as diffuse features, confined to certain stratigraphic levels, and localized within fault zones. Arrays formed by layer-bounded, mutually orthogonal sets of steeply dipping gypsum veins are reported for the first time. Vein length, height and spacing depend on the thickness of the bed packages in which they are confined. Within fault zones, veins are partly a product of faulting but are also inherited layer-bounded features along which faults are superimposed. Owing to the different petrophysical properties with respect to the parent rocks and their susceptibility to textural and mineralogical modifications, water dissolution and rupture, gypsum veins may have a significant role in geofluid management. Depending on their patterns and grade of physical and chemical alteration, veins may influence geofluid circulation and storage, acting as barriers to flow and possibly also as conduits.
A 2.3 million year lacustrine record of orbital forcing from the Devonian of northern Scotland
Authors: Andrews S. D. , Cornwell D. G. , Trewin N. H. , Hartley A. J. , Archer S. G.
Published: February 2016

Detailed sedimentological examination of onshore sections through cyclic lacustrine deposits of the Middle Devonian succession of northern Scotland has been augmented with offshore data to allow the construction of a continuous 2.3 myr record of orbital forcing. These data provide an important record of climatic variance through the Devonian, which, by comparison with other records from this time, can significantly improve our understanding of the Devonian global climatic system. Precessional and eccentricity cycles are shown to be dominant by the direct measurement and extrapolation of depositional rates in well-exposed sections as well as the analysis of ratios between primary and modulating cycles. Spectral analysis has confirmed the presence of regular cycles throughout the offshore data, which provide a far longer record of orbitally forced cyclicity than previously available. The presence of a half precession cycle is also suggested. Similar up-section variations in cycle thicknesses are recorded from both the onshore and offshore succession. These variations and those in cycle symmetry are related to a trend from an underfilled to a balanced-fill basin state. Of further importance in controlling cycle thickness is the period of each cycle during which lacustrine conditions, and therefore lake level controlled accommodation, existed.
Importance of pre-existing fault size for the evolution of an inverted fault system
Authors: Reilly Cathal , Nicol Andrew , Walsh John
Published: February 2016

Fault inversion has been documented in many basins worldwide, yet the details of how the initial extensional faults impact on the geometry and growth of the reactivated contractional system is often poorly resolved by the available data. Two-dimensional (2D) and 3D seismic reflection, and well data have been used to chart the evolution of inverted faults from the Taranaki Basin, offshore New Zealand. Sedimentary rocks up to 8 km thick record Late Cretaceous–Paleocene normal faults inverted during Miocene and younger shortening. The displacement and length of early normal faults is a key determinant for the reactivation and size of the subsequent reverse faults. All normal faults with maximum vertical displacements ≥600 m and lengths ≥9 km were inverted along their entire length, while smaller faults were not inverted. The proportion of the total basin-wide strain accommodated on each fault is comparable between deformational episodes. The hierarchy of reverse fault lengths was established rapidly, with longer faults accruing a greater proportion of the total strain from an early stage of shortening. The reverse fault system is dominated by inverted normal faults, which accrue displacement at the expense of smaller faults, and utilize the largest crustal-scale elements of the pre-existing system. The size of pre-existing heterogeneities is an important control for the magnitude and spatial extent of elevated stresses during contraction, which, in turn, control the dimensions, locations and displacements of subsequent fault growth.
The geometry and dimensions of fault-core lenses
Authors: Gabrielsen Roy H. , Braathen Alvar , Kjemperud Magnus , Valdresbråten Marie Lovise R.
Published: February 2016

Field analysis shows that fault cores of brittle, extensional faults at a medium to mature stage of development are commonly dominated by lozenge-shaped horses (fault-core lenses) characterized by a variety of lithologies, including intact, mildly to strongly deformed country rock derived from the footwalls and hanging walls, various types of fault rocks of the protocatalasite and breccia series, breccia, fault gouge and clay smear. The lenses are sometimes stacked to form complex duplexes. These structures are commonly separated by high-strain zones of sheared cataclasite, and/or clay smear/clay gouge. The geometry and distribution of clay gouge in high-strain zones sometimes display evidence of intrusion, indicating high fluid pressure.

Although the sizes of the horses vary over several orders of magnitude, they frequently display a length:thickness (
a
:
c
) ratio of between 1:4 and 1:15.

The high-strain zones of fault rocks commonly constitute unbroken, 3D membranes that are likely to constrain fluid communication both across and along the fault zone.

There are significant contrasts in fault core architecture that are probably related to processes associated with contrasting fluid pressure, strain intensity and strain hardening/strain softening. Faults associated with strain softening are characterized by less abundant brittle deformation products and are less likely to be conduits for fluid flow compared to those that are affected by strain hardening.
Post-Caledonian extension in the West Norway–northern North Sea region: the role of structural inheritance
Authors: Fossen Haakon , Khani Hamed Fazli , Faleide Jan Inge , Ksienzyk Anna K. , Dunlap W. James
Published: February 2016

The northern North Sea region has experienced repeated phases of post-Caledonian extension, starting with extensional reactivation of the low-angle basal Caledonian thrust zone, then the formation of Devonian extensional shear zones with 10–100 km-scale displacements, followed by brittle reactivation and the creation of a plethora of extensional faults. The North Sea Rift-related approximately east–west extension created a new set of rift-parallel faults that cut across less favourably orientated pre-rift structures. Nevertheless, fault rock dating shows that onshore faults and shear zones of different orientations were active throughout the history of rifting. Several of the reactivated major Devonian extensional structures can be extrapolated offshore into the rift, where they appear as bands of dipping reflectors. They coincide with large-scale boundaries separating 50–100 km-wide rift domains of internally uniform fault patterns. Major north–south-trending rift faults, such as the Øygarden Fault System, bend or terminate against these boundaries, clearly influenced by their presence during rifting. Hence, the North Sea is one of several examples where pre-rift basement structures oblique to the rift extension direction can significantly influence rift architecture, even if most of the rift faults are newly-formed structures.
Spatial characteristics of the Pliocene to modern alluvial fan successions in the uplifted sedimentary basins of Almería, SE Spain: review and regional synthesis
Authors: M. Harvey Adrian , Stokes Martin , Mather Anne , Whitfield Elizabeth
Published: February 2016

Conventional sedimentological models of alluvial fan sequences tend to be based on fans in subsiding sedimentary basins. Fans in uplifting terrains are rather different. We examined the sequence of fan delta and alluvial fan deposition from the Pliocene to the Late Quaternary in the uplifting terrain of the Neogene sedimentary basins of Almería, Betic Cordillera, SE Spain. There were three phases of fan deposition, each yielding particular morpho-sedimentological characteristics and each having distinctive spatial distributions. The first phase was the Pliocene fan deltas, deposited where the larger river systems entered the receding early Pliocene seas. This phase was followed during the late Pliocene–Early Pleistocene by a phase of extensive alluvial fan deposition extending from the mountain fronts to the basin centres. Continued uplift occurred during the Pleistocene, resulting in incision of the drainage network and dissection of the Plio-Pleistocene fan systems. The third phase, during the Quaternary, involved much smaller mountain front alluvial fans set well below the Plio-Pleistocene fans, except in local areas of tectonic subsidence where extensive fans were formed. The older fans exhibit stacked sedimentary successions with very little evidence of sustained vertical incision. However, the Quaternary fans show much evidence of sustained incision between major aggradational phases, often resulting in inset rather than stacked stratigraphic relationships. We interpret this to imply that, although the tectonic context was the primary temporal control on variations within the older fan successions, within the Quaternary fans the overall tectonic control has been modified by responses to the Quaternary climatic sequence.
Thrombin stimulates albumin transcytosis in lung microvascular endothelial cells via activation of acid sphingomyelinase
Authors: Kuebler Wolfgang M. , Wittenberg Claudia , Lee Warren L. , Reppien Eike , Goldenberg Neil M. , Lindner Karsten , Gao Yizhuo , Winoto-Morbach Supandi , Drab Marek , Mühlfeld Christian , Dombrowsky Heike , Ochs Matthias , Schütze Stefan , Uhlig Stefan
Published: February 2016

Transcellular albumin transport occurs via caveolae that are abundant in lung microvascular endothelial cells. Stimulation of albumin transcytosis by proinflammatory mediators may contribute to alveolar protein leak in lung injury, yet the regulation of albumin transport and its underlying molecular mechanisms are so far incompletely understood. Here we tested the hypothesis that thrombin may stimulate transcellular albumin transport across lung microvascular endothelial cells in an acid-sphingomyelinase dependent manner. Thrombin increased the transport of fluorescently labeled albumin across confluent human lung microvascular endothelial cell (HMVEC-L) monolayers to an extent that markedly exceeds the rate of passive diffusion. Thrombin activated acid sphingomyelinase (ASM) and increased ceramide production in HMVEC-L, but not in bovine pulmonary artery cells, which showed little albumin transport in response to thrombin. Thrombin increased total caveolin-1 (cav-1) content in both whole cell lysates and lipid rafts from HMVEC-L, and this effect was blocked by inhibition of ASM or de novo protein biosynthesis. Thrombin-induced uptake of albumin into lung microvascular endothelial cells was confirmed in isolated-perfused lungs by real-time fluorescence imaging and electron microscopy of gold-labeled albumin. Inhibition of ASM attenuated thrombin-induced albumin transport both in confluent HMVEC-L and in intact lungs, whereas HMVEC-L treatment with exogenous ASM increased albumin transport and enriched lipid rafts in cav-1. Our findings indicate that thrombin stimulates transcellular albumin transport in an acid sphingomyelinase-dependent manner by inducing de novo synthesis of cav-1 and its recruitment to membrane lipid rafts.
Increased susceptibility of Cftr−/− mice to LPS-induced lung remodeling
Authors: Bruscia Emanuela M. , Zhang Ping-Xia , Barone Christina , Scholte Bob J. , Homer Robert , Krause Diane S. , Egan Marie E.
Published: February 2016

Cystic fibrosis (CF) is caused by homozygous mutations of the CF transmembrane conductance regulator (CFTR) Cl− channel, which result in chronic pulmonary infection and inflammation, the major cause of morbidity and mortality. Although these processes are clearly related to each other, each is likely to contribute to the pathology differently. Understanding the contribution of each of these processes to the overall pathology has been difficult, because they are usually so intimately connected. Various CF mouse models have demonstrated abnormal immune responses compared with wild-type (WT) littermates when challenged with live bacteria or bacterial products acutely. However, these studies have not investigated the consequences of persistent inflammation on lung tissue in CF mice, which may better model the lung pathology in patients. We characterized the lung pathology and immune response of Cftr−/− (CF) and Cftr+/+ (WT) mice to chronic administration of Pseudomonas aeruginosa lipopolysaccharide (LPS). We show that, after long-term repeated LPS exposure, CF mice develop an abnormal and persistent immune response, which is associated with more robust structural changes in the lung than those observed in WT mice. Although CF mice and their WT littermates develop lung pathology after chronic exposure to LPS, the inflammation and damage resolve in WT mice. However, CF mice do not recover efficiently, and, as a consequence of their chronic inflammation, CF mice are more susceptible to morphological changes and lung remodeling. This study shows that chronic inflammation alone contributes significantly to aspects of CF lung pathology.
Aging Thermal Treatment in the Inconel 725 Brazed Incorporating Tungsten Nanoparticles
Authors: Hdz-García H. M. , Muñoz-Arroyo R. , Martinez A. I. , González M. A. , Granda-Gutiérrez E. E. , Acevedo-Dávila J. L. , Castro-Roman M. , García-Vázquez F.
Published: February 2016

Fractures in blade sections of Inconel 725 were impregnated with tungsten nanoparticles and jointed by the brazing process. In order to evaluate their effect over the microstructure, aging thermal treatments at 750°C for 2, 6, 10, and 14 h were done. BNi-9 was selected as brazing filler metal and was characterized by scanning electron microscopy and X-ray fluorescence. Before brazing, the fractures were impregnated with a mixture of tungsten NPs in ethanol. Measurements of Vickers microhardness showed an increase in the melting zone of samples with aging thermal treatment for 14 h, which is attributed to the precipitation of theγ′ phase with a typical size ofca. 100 nm. Likewise, the tungsten NPs modified the size and morphology of Cr-Ni eutectics into finer and uniformly distributed microstructures.
Circulating Estrogens and Postmenopausal Ovarian Cancer Risk in the Women's Health Initiative Observational Study
Authors: Trabert Britton , Brinton Louise A. , Anderson Garnet L. , Pfeiffer Ruth M. , Falk Roni T. , Strickler Howard D. , Sliesoraitis Sarunas , Kuller Lewis H. , Gass Margery L. , Fuhrman Barbara J. , Xu Xia , Wentzensen Nicolas
Published: February 2016

Background: Hormonal and reproductive factors contribute to the development of ovarian cancer, but few studies have examined associations between circulating estrogens and estrogen metabolites and ovarian cancer risk. We evaluated whether serum estrogens and estrogen metabolite levels are associated with ovarian cancer risk among postmenopausal women in a nested case–control study in the Women's Health Initiative (WHI) Observational Study (OS).

Methods: We selected all 169 eligible epithelial ovarian cancer cases and 412 matched controls from women enrolled in WHI-OS who were not using menopausal hormones at baseline. Baseline levels of 15 estrogens and estrogen metabolites were measured via liquid chromatography/tandem mass spectrometry. Associations with ovarian cancer risk overall and stratified by histologic subtype (serous/nonserous) were analyzed using logistic regression. The mean time from serum collection to cancer diagnosis was 6.9 years.

Results: Overall, we observed modest ovarian cancer risk associations among women with higher levels of estrone [OR (95% confidence interval) quintile (Q)5 vs. Q1: 1.54 (0.82–2.90), Ptrend = 0.05], as well as 2- and 4-methoxyestrone metabolites [2.03 (1.06–3.88), Ptrend = 0.02; 1.86 (0.98–3.56), Ptrend = 0.01, respectively]. Associations of estrogens and estrogen metabolites varied substantially by histologic subtype. Associations with serous tumors were universally null, while estrone [2.65 (1.09–6.45), Ptrend = 0.01, Pheterogeneity = 0.04], unconjugated estradiol [2.72 (1.04–7.14), Ptrend = 0.03, Pheterogeneity = 0.02] and many of the 2-, 4-, and 16-pathway metabolites were positively associated with nonserous tumors.

Conclusions: Our study provides novel molecular data showing an association of the parent estrogens and several estrogen metabolites with nonserous ovarian cancers.

Impact: These findings further support the heterogeneous etiology of ovarian cancer. Cancer Epidemiol Biomarkers Prev; 25(4); 648–56. ©2016 AACR.
A Randomized Phase II/III Study of Naptumomab Estafenatox + IFNα versus IFNα in Renal Cell Carcinoma: Final Analysis with Baseline Biomarker Subgroup and Trend Analysis
Authors: Hawkins Robert E. , Gore Martin , Shparyk Yaroslav , Bondar Vladimir , Gladkov Oleg , Ganev Tosho , Harza Mihai , Polenkov Serhii , Bondarenko Igor , Karlov Petr , Karyakin Oleg , Khasanov Rustem , Hedlund Gunnar , Forsberg Goran , Nordle Örjan , Eisen Timothy
Published: February 2016

Purpose: To prospectively determine the efficacy of naptumomab estafenatox (Nap) + IFNα versus IFN in metastatic renal cell carcinoma (RCC).

Experimental Design: In a randomized, open-label, multicenter, phase II/III study, 513 patients with RCC received Nap (15 μg/kg i. v. in three cycles of four once-daily injections) + IFN (9 MU s.c. three times weekly), or the same regimen of IFN monotherapy. The primary endpoint was overall survival (OS).

Results: This phase II/III study did not meet its primary endpoint. Median OS/PFS for Nap + IFN patients was 17.1/5.8 months versus 17.5/5.8 months for the patients receiving IFN alone (P = 0.56; HR, 1.08/P = 0.41; HR, 0.92). Post hoc exploratory subgroup and trend analysis revealed that the baseline plasma concentrations of anti-SEA/E-120 (anti-Nap antibodies) for drug exposure and IL6 for immune status could be used as predictive biomarkers. A subgroup of patients (SG; n = 130) having concentrations below median of anti-SEA/E-120 and IL6 benefitted greatly from the addition of Nap. In SG, median OS/PFS for the patients treated with Nap + IFN was 63.3/13.7 months versus 31.1/5.8 months for the patients receiving IFN alone (P = 0.02; HR, 0.59/P = 0.02; HR, 0.62). Addition of Nap to IFN showed predicted and transient immune related AEs and the treatment had an acceptable safety profile.

Conclusions: The study did not meet its primary endpoint. Nap + IFN has an acceptable safety profile, and results from post hoc subgroup analyses showed that the treatment might improve OS/PFS in a baseline biomarker-defined RCC patient subgroup. The results warrant further studies with Nap in this subgroup. Clin Cancer Res; 22(13); 3172–81. ©2016 AACR.
Epithelial–Mesenchymal Transition Is Associated with a Distinct Tumor Microenvironment Including Elevation of Inflammatory Signals and Multiple Immune Checkpoints in Lung Adenocarcinoma
Authors: Lou Yanyan , Diao Lixia , Cuentas Edwin Roger Parra , Denning Warren L. , Chen Limo , Fan You Hong , Byers Lauren A. , Wang Jing , Papadimitrakopoulou Vassiliki A. , Behrens Carmen , Rodriguez Jaime Canales , Hwu Patrick , Wistuba Ignacio I. , Heymach John V. , Gibbons Don L.
Published: February 2016

Purpose: Promising results in the treatment of non–small cell lung cancer (NSCLC) have been seen with agents targeting immune checkpoints, such as programmed cell death 1 (PD-1) or programmed death ligand-1 (PD-L1). However, only a select group of patients respond to these interventions. The identification of biomarkers that predict clinical benefit to immune checkpoint blockade is critical to successful clinical translation of these agents.

Methods: We conducted an integrated analysis of three independent large datasets, including The Cancer Genome Atlas of lung adenocarcinoma and two datasets from MD Anderson Cancer Center (Houston, TX), Profiling of Resistance Patterns and Oncogenic Signaling Pathways in Evaluation of Cancers of the Thorax (named PROSPECT) and Biomarker-Integrated Approaches of Targeted Therapy for Lung Cancer Elimination (named BATTLE-1). Comprehensive analysis of mRNA gene expression, reverse-phase protein array, IHC, and correlation with clinical data were performed.

Results: Epithelial–mesenchymal transition (EMT) is highly associated with an inflammatory tumor microenvironment in lung adenocarcinoma, independent of tumor mutational burden. We found immune activation coexistent with elevation of multiple targetable immune checkpoint molecules, including PD-L1, PD-L2, PD-1, TIM-3, B7-H3, BTLA, and CTLA-4, along with increases in tumor infiltration by CD4+Foxp3+ regulatory T cells in lung adenocarcinomas that displayed an EMT phenotype. Furthermore, we identify B7-H3 as a prognostic marker for NSCLC.

Conclusions: The strong association between EMT status and an inflammatory tumor microenvironment with elevation of multiple targetable immune checkpoint molecules warrants further investigation of using EMT as a predictive biomarker for immune checkpoint blockade agents and other immunotherapies in NSCLC and possibly a broad range of other cancers. Clin Cancer Res; 22(14); 3630–42. ©2016 AACR.

See related commentary by Datar and Schalper, p. 3422
STAT3/5-Dependent IL9 Overexpression Contributes to Neoplastic Cell Survival in Mycosis Fungoides
Authors: Vieyra-Garcia Pablo A. , Wei Tianling , Naym David Gram , Fredholm Simon , Fink-Puches Regina , Cerroni Lorenzo , Odum Niels , O'Malley John T. , Gniadecki Robert , Wolf Peter
Published: February 2016

Purpose: Sustained inflammation is a key feature of mycosis fungoides (MF), the most common form of cutaneous T-cell lymphoma (CTCL). Resident IL9–producing T cells have been found in skin infections and certain inflammatory skin diseases, but their role in MF is currently unknown.

Experimental Design: We analyzed lesional skin from patients with MF for the expression of IL9 and its regulators. To determine which cells were producing IL9, high-throughput sequencing was used to identify malignant clones and Vb-specific antibodies were employed to visualize malignant cells in histologic preparations. To explore the mechanism of IL9 secretion, we knocked down STAT3/5 and IRF4 by siRNA transfection in CTCL cell lines receiving psoralen+UVA (PUVA) ± anti-IL9 antibody. To further examine the role of IL9 in tumor development, the EL-4 T-cell lymphoma model was used in C57BL/6 mice.

Results: Malignant and reactive T cells produce IL9 in lesional skin. Expression of the Th9 transcription factor IRF4 in malignant cells was heterogeneous, whereas reactive T cells expressed it uniformly. PUVA or UVB phototherapy diminished the frequencies of IL9- and IL9r-positive cells, as well as STAT3/5a and IRF4 expression in lesional skin. IL9 production was regulated by STAT3/5 and silencing of STAT5 or blockade of IL9 with neutralizing antibodies potentiated cell death after PUVA treatment in vitro. IL9-depleted mice exhibited a reduction of tumor growth, higher frequencies of regulatory T cells, and activated CD4 and CD8 T lymphocytes.

Conclusions: Our results suggest that IL9 and its regulators are promising new targets for therapy development in mycosis fungoides. Clin Cancer Res; 22(13); 3328–39. ©2016 AACR.
miR-155 Overexpression Promotes Genomic Instability by Reducing High-fidelity Polymerase Delta Expression and Activating Error-Prone DSB Repair
Authors: Czochor Jennifer R. , Sulkowski Parker , Glazer Peter M.
Published: February 2016

miR-155 is an oncogenic miRNA that is often overexpressed in cancer and is associated with poor prognosis. miR-155 can target several DNA repair factors, including RAD51, MLH1, and MSH6, and its overexpression results in an increased mutation frequency in vitro, although the mechanism has yet to be fully understood. Here, we demonstrate that overexpression of miR-155 drives an increased mutation frequency both in vitro and in vivo, promoting genomic instability by affecting multiple DNA repair pathways. miR-155 overexpression causes a decrease in homologous recombination, but yields a concurrent increase in the error-prone nonhomologous end-joining pathway. Despite repressing established targets MLH1 and MSH6, the identified mutation pattern upon miR-155 overexpression does not resemble that of a mismatch repair–deficient background. Further investigation revealed that all four subunits of polymerase delta, a high-fidelity DNA replication, and repair polymerase are downregulated at the mRNA level in the context of miR-155 overexpression. FOXO3a, a transcription factor and known target of miR-155, has one or more putative binding site(s) in the promoter of all four polymerase delta subunits. Finally, suppression of FOXO3a by miR-155 or by siRNA knockdown is sufficient to repress the expression of the catalytic subunit of polymerase delta, POLD1, at the protein level, indicating that FOXO3a contributes to the regulation of polymerase delta levels.

Implications: Taken together, miR-155 overexpression drives an increase in mutation frequency via multifaceted impact on DNA damage response and DNA repair pathways. Mol Cancer Res; 14(4); 363–73. ©2016 AACR.
A Computational Modeling Approach for Deriving Biomarkers to Predict Cancer Risk in Premalignant Disease
Authors: Dhawan Andrew , Graham Trevor A. , Fletcher Alexander G.
Published: February 2016

The lack of effective biomarkers for predicting cancer risk in premalignant disease is a major clinical problem. There is a near-limitless list of candidate biomarkers, and it remains unclear how best to sample the tissue in space and time. Practical constraints mean that only a few of these candidate biomarker strategies can be evaluated empirically, and there is no framework to determine which of the plethora of possibilities is the most promising. Here, we have sought to solve this problem by developing a theoretical platform for in silico biomarker development. We construct a simple computational model of carcinogenesis in premalignant disease and use the model to evaluate an extensive list of tissue sampling strategies and different molecular measures of these samples. Our model predicts that (i) taking more biopsies improves prognostication, but with diminishing returns for each additional biopsy; (ii) longitudinally collected biopsies provide slightly more prognostic information than a single biopsy collected at the latest possible time point; (iii) measurements of clonal diversity are more prognostic than measurements of the presence or absence of a particular abnormality and are particularly robust to confounding by tissue sampling; and (iv) the spatial pattern of clonal expansions is a particularly prognostic measure. This study demonstrates how the use of a mechanistic framework provided by computational modeling can diminish empirical constraints on biomarker development. Cancer Prev Res; 9(4); 283–95. ©2016 AACR.
Hepatic Premalignant Alterations Triggered by Human Nephrotoxin Aristolochic Acid I in Canines
Authors: Jin Ke , Su Kun-kai , Li Tong , Zhu Xia-qing , Wang Qi , Ge Ren-shan , Pan Zong-fu , Wu Bo-wen , Ge Li-jun , Zhang Yi-han , Wang Yi-fan , Shen Guo-fang , Zhu Dan-yan , Xiang Chun-sheng , Li Lan-juan , Lou Yi-jia
Published: February 2016

Aristolochic acid I (AAI) existing in plant drugs from Aristolochia species is an environmental human carcinogen associated with urothelial cancer. Although gene association network analysis demonstrated gene expression profile changes in the liver of human TP53 knock-in mice after acute AAI exposure, to date, whether AAI causes hepatic tumorigenesis is still not confirmed. Here, we show that hepatic premalignant alterations appeared in canines after a 10-day AAI oral administration (3 mg/kg/day). We observed c-Myc oncoprotein and oncofetal RNA-binding protein Lin28B overexpressions accompanied by cancer progenitor-like cell formation in the liver by AAI exposure. Meanwhile, we found that forkhead box O1 (FOXO1) was robustly phosphorylated, thereby shuttling into the cytoplasm of hepatocytes. Furthermore, utilizing microarray and qRT-PCR analysis, we confirmed that microRNA expression significantly dysregulated in the liver treated with AAI. Among them, we particularly focused on the members in let-7 miRNAs and miR-23a clusters, the downstream of c-Myc and IL6 receptor (IL6R) signaling pathway linking the premalignant alteration. Strikingly, when IL6 was added in vitro, IL6R/NF-κB signaling activation contributed to the increase of FOXO1 phosphorylation by the let-7b inhibitor. Therefore, it highlights the new insight into the interplay of the network in hepatic tumorigenesis by AAI exposure, and also suggests that anti-premalignant therapy may be crucial for preventing AAI-induced hepatocarcinogenesis. Cancer Prev Res; 9(4); 324–34. ©2016 AACR.
Characterizing the Molecular Pathology of Arrhythmogenic Cardiomyopathy in Patient Buccal Mucosa Cells
Authors: Asimaki Angeliki , Protonotarios Alexandros , James Cynthia A. , Chelko Stephen P. , Tichnell Crystal , Murray Brittney , Tsatsopoulou Adalena , Anastasakis Aris , te Riele Anneline , Kléber André G. , Judge Daniel P. , Calkins Hugh , Saffitz Jeffrey E.
Published: February 2016

Background—

Analysis of myocardium has revealed mechanistic insights into arrhythmogenic cardiomyopathy but cardiac samples are difficult to obtain from probands and especially from family members. To identify a potential surrogate tissue, we characterized buccal mucosa cells.

Methods and Results—

Buccal cells from patients, mutation carriers, and controls were immunostained and analyzed in a blinded fashion. In additional studies, buccal cells were grown in vitro and incubated with SB216763. Immunoreactive signals for the desmosomal protein plakoglobin and the major cardiac gap junction protein Cx43 were markedly diminished in buccal mucosa cells from arrhythmogenic cardiomyopathy patients with known desmosomal mutations when compared with controls. Plakoglobin and Cx43 signals were also reduced in most family members who carried disease alleles but showed no evidence of heart disease. Signal for the desmosomal protein plakophilin-1 was reduced in buccal mucosa cells in patients with
PKP2
mutations but not in those with mutations in other desmosomal genes. Signal for the desmosomal protein desmoplakin was reduced in buccal mucosa cells from patients with mutations in
DSP
,
DSG2
, or
DSC2
but not in
PKP2
or
JUP
. Abnormal protein distributions were reversed in cultured cells incubated with SB216763, a small molecule that rescues the disease phenotype in cardiac myocytes.

Conclusions—

Buccal mucosa cells from arrhythmogenic cardiomyopathy patients exhibit changes in the distribution of cell junction proteins similar to those seen in the heart. These cells may prove useful in future studies of disease mechanisms and drug screens for effective therapies in arrhythmogenic cardiomyopathy.
The Secession of Crimea: Where Does International Law Stand?
Authors: Nikouei Majid , Zamani Masoud
Published: February 2016

There is more legal complexity to the secession of Crimea than meets the eye upon the first encounter with the 2014–2015 crisis of Ukraine. The secession of Crimea calls into question the traditional configuration of the right to self-determination. The question of Crimea is compounded when, in discerning the legality of some of the key issues linked to it, one must pay regard to the standards of both international law and national law. In other words, a constructive analysis of the secession of Crimea must delve into the national law standards of Ukraine, whilst simultaneously analysing the effect of observing or otherwise ignoring those standards as for the legality of secession therein from the perspective of international law. With a view to this complexity, this essay aims to combine some of the most controversial dimensions to the secession of Crimea in one piece, and clarify whether the case of Crimea can in any way be justified by the rules of international law on self-determination.
The Siren’s Call? Exploring the Implications of an Additional Protocol to the European Convention on Human Rights on National Minorities
Authors: Berry Stephanie E.
Published: February 2016

Calls for the adoption of an Additional Protocol to the European Convention on Human Rights (echr) on National Minorities have persisted within the Council of Europe despite the adoption of the Framework Convention for the Protection of National Minorities (fcnm). This article explores the potential implications of the adoption of an Additional Protocol on National Minorities to the echr for the fcnm. The European Court of Human Rights (ECtHR) already has several tools that would allow it to extend protection to persons belonging to national minorities. However, as the ECtHR tends to allow States a wide margin of appreciation in cases concerning persons belonging to minorities, it is argued that the adoption of an Additional Protocol on National Minorities may not be desirable, as it has the potential to undermine the progress made by the Framework Convention Advisory Committee.