bluuebunny/crossref_metadata_2025 huggingface dataset:
Military intelligence
Authors: Rankov Boris
Published: December 2015

Strategic intelligence‐gathering in the ancient Mediterranean relied onproxenoi(representatives) and ambassadors, who could act as agents and spies, and on the passing of information by allies and traders, although this was not orchestrated or analyzed by specialist agencies. Its value was limited by slowness of communications, which meant that it was often out of date before any response could be brought to bear. Tactical intelligence‐gathering on the march or on the battlefield was more effective, and relied on cavalry screens, scouts, and sometimes spies at night and when close to the enemy. Prisoners, deserters, and refugees could also be interrogated to provide corroboration of information already collected.
Plutarch of Athens
Authors: Chiaradonna Riccardo
Published: December 2015

Plutarch of Athens (d. 432
ce
) was a Neoplatonic philosopher active in the late fourth and the early fifth century
ce
. He was head of the Platonic school in Athens, which flourished until 529
ce
. Among his students were the Neoplatonic philosophers Syrianus, Hierocles of Alexandria, and Proclus. Plutarch's works are lost. Fragments from his commentary on Aristotle's
On the soul
have been preserved by later Neoplatonic exegetes. He also wrote commentaries on Plato's dialogues, most notably the
Parmenides
. Plutarch is a key figure who shaped late antique Platonism by developing Iamblichus’ philosophical and theological teaching.
Portugal
Authors: Gosner Linda
Published: December 2015

Portugal is located in southwest Europe and together with Spain makes up the Iberian peninsula, which divides the Atlantic Ocean from the Mediterranean Sea. The country has an ecologically diverse landscape ranging from a mountainous, wet, cool environment in the north to rolling plains and a Mediterranean climate in the south. It is rich in metals, agricultural land, and marine resources, which were all exploited in antiquity. During the Bronze Age, stockbreeding, agriculture, mining, and interregional connections increased. The Iron Age is characterized by increased contact with Phoenician and Punic colonizers as well as the proliferation of the Tartessian culture in the south and the development of large, fortified
castro
settlements in the north. Over the course of the final two centuries
bce
, Portugal came under Roman control. It became part of Roman Hispania and was governed as the province of Lusitania from the capital, Augusta Emerita (Mérida).
Religion in contact, Iberian peninsula
Authors: Marco Francisco
Published: December 2015

In the Iberian peninsula there were two major linguistic zones: one inhabited by peoples who did not speak Indo‐European languages, known as “Iberians,” along the coast from lower Andalusia to Languedoc (with earlier Mediterranean influences), and the interior inhabited by the Hispano‐Celts. While the Orientalizing period is defined by the “princely graves,” from the fourth century
bce
onward there is a shift to the sanctuaries, where a new collective image of society is expressed through bronze figurines, stone sculpture, or pottery vessels. With the selective adoption of Italo‐Hellenistic conventions, a process of monumentalization is attested in the second and first centuries
bce
, and the names of local deities or rituals emerge in the epigraphy of the Celtic interior.
Sicily, Phoenician
Authors: Urquhart Lela M.
Published: December 2015

Phoenician presence in Sicily dates back to the eighth century and is documented for Motya, Panormus, and Soloeis. The majority of evidence is archaeological, dating to between the eighth and fourth century
bce
, and relates to different aspects of Phoenician culture, including funerary customs, religion, craftwork, and commerce.
Struggle of the Orders
Authors: Forsythe Gary
Published: December 2015

Modern scholars use this term to characterize a common theme in ancient accounts of early Roman history that describe affairs as driven by a struggle between the two orders or classes: patricians and plebeians. Hereby ancient writers explain virtually all innovations in the Roman state during the Early Republic (sixth to third century). The narrative of the Struggle of the Orders was clearly influenced by the history of the Late Republic, involving clashes between populist aristocratic leaders and other aristocrats heading more conservative followings. But modern scholars differ concerning to what extent ancient historiographers have falsified Early Republican history through Late Republican anachronisms.
Tel Kabri
Authors: Vacek Alexander
Published: December 2015

Tel Kabri is located in the eastern part of the western Galilee plain. A substantial Bronze Age palace was recovered at the site, which finds its closest parallel in Alalakh phase VII. During the Iron Age parts of the Tel were occupied by a fortress, which housed a garrison of Greek mercenaries in the seventh century
bce
.
Valencia–Murcia
Authors: Grau‐Mira Ignasi
Published: December 2015

This entry describes the main political processes that occurred in eastern Iberia from the Iron Age to the Roman Empire. Located in this area were some of the most important Iberian peoples, such as the Contestanians and Edetanians. These peoples developed a major urban culture that was the basis of Roman settlement in the region.
A new differential pressure flow meter for measurement of human breath flow: Simulation and experimental investigation
Authors: Bridgeman Devon , Tsow Francis , Xian Xiaojun , Forzani Erica
Published: December 2015

The development and performance characterization of a new differential pressure‐based flow meter for human breath measurements is presented in this article. The device, called a “Confined Pitot Tube,” is comprised of a pipe with an elliptically shaped expansion cavity located in the pipe center, and an elliptical disk inside the expansion cavity. The elliptical disk, named Pitot Tube, is exchangeable, and has different diameters, which are smaller than the diameter of the elliptical cavity. The gap between the disk and the cavity allows the flow of human breath to pass through. The disk causes an obstruction in the flow inside the pipe, but the elliptical cavity provides an expansion for the flow to circulate around the disk, decreasing the overall flow resistance. We characterize the new sensor flow experimentally and theoretically, using Comsol Multiphysics® software with laminar and turbulent models. We also validate the sensor, using inhalation and exhalation tests and a reference method. © 2016 American Institute of Chemical Engineers AIChE J, 62: 956–964, 2016
Production of benzene, toluene, and xylenes from natural gas via methanol: Process synthesis and global optimization
Authors: Niziolek Alexander M. , Onel Onur , Floudas Christodoulos A.
Published: December 2015

A systematic global optimization‐based process synthesis framework is presented to determine the most profitable processes to produce aromatics from natural gas. Several novel, commercial, and/or competing technologies are modeled within the framework, including methanol‐to‐aromatics, toluene alkylation with methanol, selective toluene disproportionation, and toluene disproportionation and transalkylation with heavy aromatics, among others. We propose a stand‐alone chemicals facility: the main products are aromatics with allowable by‐products of gasoline, liquefied petroleum gas, and electricity. Several case studies are discussed that produce varying ratios of para‐, ortho‐, and meta‐xylene across multiple refinery capacities. The results indicate that utilizing natural gas for the production of aromatics is profitable with net present values as high as $3800 MM dollars and payback periods as low as 6 years. The required investment for these refineries represents as much as a 65% decrease compared to published estimates of similar coal‐based capacity plants. © 2016 American Institute of Chemical Engineers AIChE J, 62: 1531–1556, 2016
Toward efficient water/oil separation material: Effect of copolymer composition on pH‐responsive wettability and separation performance
Authors: Zhou Yin‐Ning , Li Jin‐Jin , Luo Zheng‐Hong
Published: December 2015

Interest in functional soft matter with stimuli‐responsive wettability has increasingly intensified in recent years. From the chemical product engineering viewpoint, this study aims to fabricate reversible pH‐responsive polymeric surfaces with controllable wettability using [poly(2,2,3,4,4,4‐hexafluorobutyl methacrylate)‐block‐ poly(acrylic acid) (PHFBMA‐b‐PAA)] block copolymers. To attain this aim, three block copolymers with different PAA segment lengths were synthesized for the first time through Cu(0)‐mediated reversible‐deactivation radical polymerization and hydrolysis reaction. pH‐induced controllable wettability was achieved by spin‐coating the resulting block copolymers onto silicon wafers. Results showed that the pH‐responsive wetting behavior was introduced by incorporating the PAA block, and that the responsiveness of as‐fabricated surfaces was greatly influenced by PAA content. All three evolutions of water contact angle with pH shared a similar inflection point at pH 5.25. Furthermore, on the basis of the wetting properties and mechanism understanding, the application of copolymer coated meshes in layered water/oil separation was exploited. Given their superhydrophilicity and underwater superoleophobicity, PHFBMA70‐b‐PAA148 and PHFBMA70‐b‐PAA211 coated stainless steel meshes (SSMs) can efficiently separate water from different mixtures of organic solvent and water with high flux. However, considering long‐term use, the PHFBMA70‐b‐PAA148 coated SSM with good stability may be the best copolymer for water/oil separation. Therefore, a coordination of structure, composition, and functionality was necessary to enable practical applications of the functional materials. © 2016 American Institute of Chemical Engineers AIChE J, 62: 1758–1771, 2016
Digit ratio (2D:4D): Is it possible to use it for sex determination in the study of human skeletal remains?
Authors: Bakholdina Varvara Yu. , Movsesian Alla A. , Sineva Irina M.
Published: December 2015

Objectives

Sexual dimorphism in the relative length of the second‐to‐fourth digits (the digit ratio, or 2D:4D) in humans has been reported in many studies. The aim of our study was to ascertain possibility of using the 2D:4D ratio as an additional marker for sex determination in the study of human skeletal remains.

Methods

We have studied 2D:4D ratios obtained from measurements of finger phalanges and metacarpal bones in Russian (45 adult males and 26 adult females) and German (58 adult males and 29 adult females) skeletal series.

Results

The difference in 2D:4D ratio between the male and female subsamples in both skeletal series was not statistically significant. Analysis of variance revealed that the 2D:4D ratios in our sample varied more by ethnicity than by the sexual identity of the skeletal material.

Conclusion

Our results suggest that the 2D:4D ratio cannot be used as an appropriate trait for the sex determination of human skeletal remains. Am. J. Hum. Biol. 28:591–593, 2016. © 2015 Wiley Periodicals, Inc.
Duplications upstream and downstream of SHOX identified as novel causes of Leri–Weill dyschondrosteosis or idiopathic short stature
Authors: Bunyan David J. , Baffico Maria , Capone Lucia , Vannelli Silvia , Iughetti Lorenzo , Schmitt Sébastien , Taylor Emma‐Jane , Herridge Adam A. , Shears Deborah , Forabosco Antonino , Coviello Domenico A.
Published: December 2015

Leri–Weill dyschondrosteosis is a pseudoautosomal dominantly‐inherited skeletal dysplasia ascribed to haploinsufficiency of the SHOX gene caused by deletions, point mutations, or partial duplications of the gene, or to heterozygous deletions upstream or downstream of the intact SHOX gene involving conserved non‐coding cis‐regulatory DNA elements that show enhancer activity. Recently, two SHOX conserved non‐coding element duplications, one upstream and one downstream, were reported in patients referred with idiopathic short stature. To further evaluate the role of these duplications in SHOX‐related disorders, we describe seven patients (five with Leri‐Weill dyschondrosteosis and two with short stature) all of whom have duplications of part of the upstream or downstream conserved non‐coding element regions, identified by multiplex ligation‐dependent probe amplification. In addition, we show data from 32 patients with an apparently identical downstream duplication that includes a proposed putative regulatory element (identified by multiplex ligation‐dependent probe amplification or array comparative genome hybridization), which results in a variable phenotype from normal to mild Leri–Weill dyschondrosteosis. These additional data provide further evidence that duplications of upstream and downstream long range cis‐regulatory DNA elements can result in a SHOX‐related phenotype. © 2015 Wiley Periodicals, Inc.
Vitamin D heritability and effect of pregnancy status in Vervet monkeys (Chlorocebus aethiops sabaeus) under conditions of modest and high dietary supplementation
Authors: Chittoor Geetha , Pajewski Nicholas M. , Voruganti V. Saroja , Comuzzie Anthony G. , Clarkson Thomas B. , Nudy Matthew , Schnatz Peter F. , Kaplan Jay R. , Jorgensen Matthew J.
Published: December 2015

Objectives

The two objectives of the current study were to: 1) investigate the genetic contributions to variations in serum vitamin D concentrations under two dietary conditions (a standard monkey biscuit diet vs. a diet designed to model typical American consumption); and 2) explore the interaction of vitamin D with pregnancy status using a cohort of pedigreed female vervet/African green monkeys.

Methods

This study includes 185 female (≥3.5 years) vervet/African green monkeys (Chlorocebus aethiops sabaeus) from a multi‐generational, pedigreed breeding colony. The 25(OH)D3 concentrations were first measured seven to eight weeks after consuming a “typical American” diet (TAD), deriving 37, 18, and 45% of calories from fat, protein sources, and carbohydrates, and supplemented with vitamin D to a human equivalent of 1,000 IU/day. Vitamin D concentrations were assessed again when animals were switched to a low‐fat, standard biscuit diet (LabDiet 5038) for 8 months, which provided a human equivalent of approximately 4,000 IU/day of vitamin D. All statistical analyses were implemented in SOLAR.

Results

Pregnancy was associated with reduced 25(OH)D3 concentrations. Heritability analyses indicated a significant genetic contribution to 25(OH)D3 concentrations in the same monkeys consuming the biscuit diet (h2=0.66, P=0.0004) and TAD (h2=0.67, P=0.0078) diets, with higher 25(OH)D3 concentrations in animals consuming the biscuit diet. Additionally, there was a significant genotype‐by‐pregnancy status interaction on 25(OH)D3 concentrations (P<0.05) only among animals consuming the TAD diet.

Discussion

These results support the existence of a genetic contribution to differences in serum 25(OH)D3 concentrations by pregnancy status and emphasize the role of diet (including vitamin D supplementation) in modifying genetic signals as well as vitamin D concentrations. Am J Phys Anthropol 159:639–645, 2016. © 2015 Wiley Periodicals, Inc.
Prevalence and determinants of periodic limb movements in the general population
Authors: Haba‐Rubio José , Marti‐Soler Helena , Marques‐Vidal Pedro , Tobback Nadia , Andries Daniela , Preisig Martin , Waeber Gérard , Vollenweider Peter , Kutalik Zoltán , Tafti Mehdi , Heinzer Raphaël
Published: December 2015

Objective

Periodic limb movements during sleep (PLMS) are sleep phenomena characterized by periodic episodes of repetitive stereotyped limb movements. The aim of this study was to describe the prevalence and determinants of PLMS in a middle to older aged general population.

Methods

Data from 2,162 subjects (51.2% women, mean age = 58.4 ± 11.1 years) participating in a population‐based study (HypnoLaus, Lausanne, Switzerland) were collected. Assessments included laboratory tests, sociodemographic data, personal and treatment history, and full polysomnography at home. PLMS index (PLMSI) was determined, and PLMSI > 15/h was considered as significant.

Results

Prevalence of PLMSI > 15/h was 28.6% (31.3% in men, 26% in women). Compared to subjects with PLMSI ≤ 15/h, subjects with PLMSI > 15/h were older (p < 0.001), were predominantly males (p = 0.007), had a higher proportion of restless legs syndrome (RLS; p < 0.001), had a higher body mass index (p = 0.001), and had a lower mean glomerular filtration rate (p < 0.001). Subjects with PLMSI > 15/h also had a higher prevalence of diabetes, hypertension, and beta‐blocker or hypnotic treatments. The prevalence of antidepressant use was higher, but not statistically significant (p = 0.07). Single nucleotide polymorphisms (SNPs) within BTBD9 (rs3923809), TOX3 (rs3104788), and MEIS1 (rs2300478) genes were significantly associated with PLSMI > 15/h. Conversely, mean hemoglobin and ferritin levels were similar in both groups. In the multivariate analysis, age, male gender, antidepressant intake, RLS, and rs3923809, rs3104788, and rs2300478 SNPs were independently associated with PLMSI > 15/h.

Interpretation

PLMS are highly prevalent in our middle‐aged European population. Age, male gender, RLS, antidepressant treatment, and specific BTBD9, TOX3, and MEIS1 SNP distribution are independent predictors of PLMSI > 15/h. ANN NEUROL 2016;79:464–474
Compound heterozygous FXN mutations and clinical outcome in friedreich ataxia
Authors: Galea Charles A. , Huq Aamira , Lockhart Paul J. , Tai Geneieve , Corben Louise A. , Yiu Eppie M. , Gurrin Lyle C. , Lynch David R. , Gelbard Sarah , Durr Alexandra , Pousset Francoise , Parkinson Michael , Labrum Robyn , Giunti Paola , Perlman Susan L. , Delatycki Martin B. , Evans‐Galea Marguerite V.
Published: December 2015

Objective

Friedreich ataxia (FRDA) is an inherited neurodegenerative disease characterized by ataxia and cardiomyopathy. Homozygous GAA trinucleotide repeat expansions in the first intron of FXN occur in 96% of affected individuals and reduce frataxin expression. Remaining individuals are compound heterozygous for a GAA expansion and a FXN point/insertion/deletion mutation. We examined disease‐causing mutations and the impact on frataxin structure/function and clinical outcome in FRDA.

Methods

We compared clinical information from 111 compound heterozygotes and 131 individuals with homozygous expansions. Frataxin mutations were examined using structural modeling, stability analyses and systematic literature review, and categorized into four groups: (1) homozygous expansions, and three compound heterozygote groups; (2) null (no frataxin produced); (3) moderate/strong impact; and (4) minimal impact. Mean age of onset and the presence of cardiomyopathy and diabetes mellitus were compared using regression analyses.

Results

Mutations in the hydrophobic core of frataxin affected stability whereas surface residue mutations affected interactions with iron sulfur cluster assembly and heme biosynthetic proteins. The null group of compound heterozygotes had significantly earlier age of onset and increased diabetes mellitus, compared to the homozygous expansion group. There were no significant differences in mean age of onset between homozygotes and the minimal and moderate/strong impact groups.

Interpretation

In compound heterozygotes, expression of partially functional mutant frataxin delays age of onset and reduces diabetes mellitus, compared to those with no frataxin expression from the non‐expanded allele. This integrated analysis of categorized frataxin mutations and their correlation with clinical outcome provide a definitive resource for investigating disease pathogenesis in FRDA. Ann Neurol 2016;79:485–495
Influence of thermal history on crystalline morphologies of isotactic polypropylene in its miscible blends with polybutene‐1
Authors: Chen Wei , Wu Lihui , Chen Liang , Qi Zeming , Li Liangbin
Published: December 2015

The miscibility behaviors in blends of isotactic polypropylene (iPP) and polybutene‐1 (PB) have been studied using in‐situ FTIR imaging. The heterogeneous melt of 3/7 iPP/PB blends were formed at 250, 220, and 180°C and then quenched to the same crystallization temperature of iPP at 125°C, respectively. Evolution processes of composition distribution during crystallization were monitored according to their characteristic peaks, and the results suggest a trend from local concentration to uniform dispersion of PB fraction. Further studies of the PB fraction as the distance from the growth front of iPP spherulite indicate an irreversible phase behavior with the progress of thermal history. The cyclic melting and crystallization favor the mixing of iPP/PB blend. Meanwhile, the nonlinear growth rate of iPP spherulite is mainly responsible for compatible promotion of iPP/PB blend, which hinders the transportation of iPP chains to its growth front. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43282.
Strong and ductile poly(butylene adipate‐co‐terephthalate) biocomposites fabricated by oscillation shear injection molding
Authors: Li Xu‐Juan , Chen Jing‐Bin , Xu Huan , Xie Lan , Zhong Gan Ji , Ran Rong , Ji Xu , Li Zhong‐Ming
Published: December 2015

Poor interfacial properties and uncontrollable phase morphology encountered during the fabrication of poly(butylene adipate‐co‐terephthalate) (PBAT)/thermal plastic starch (TPS) biocomposites, result unfortunately in low mechanical performances and thus limit its applications. Here an approach in terms of phase morphology controlling, i.e., extrusion compounding followed by oscillation shear injection molding (OSIM), is proposed to construct in situ TPS fiber and skin‐core structure consisting of TPS fiber and droplet in skin layer, and spherical TPS in core layer, which tremendously benefits the mechanical properties. Specifically, the tensile strength, modulus and ductility for the biocomposites with various loadings of TPS, even when TPS loading as high as 55 wt %, outperform pure PBAT sample fabricated by conventional injection molding (CIM) with the increment of 51%, 308% in strength and modulus, respectively. Meanwhile, the elongation at breakage can maintain at 196%. The unprecedented establishment of high‐performance PBAT/TPS biocomposites is in great need for potential applications, such as green packaging. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43312.
Properties of treated calcium copper titanate filled epoxy thin film composites for electronic applications
Authors: Satia Mohd Saidina Dandan , Jaafar Mariatti
Published: December 2015

In this study, calcium copper titanate (CCTO) fillers were treated with 1%, 5%, and 10% of silane‐based coupling agent 3‐glycidoxypropyltrimethoxysilane (GPTMS). The CCTO treated with 10% GPTMS filled epoxy thin film composite exhibits a remarkable improvement (60%) of dielectric constant than untreated CCTO/epoxy thin film composite. Besides that, results treated CCTO/epoxy thin film composite produced using various epoxy resins showed that OP 392 epoxy thin film composite exhibited the highest glass transition temperature and degradation temperature, moderate dielectric constant, slightly lower coefficient of thermal expansion and lowest dielectric loss compared with D.E.R. 332 and Epolam 2015 epoxy thin film composites. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43313.
Preparation of pseudocapacitor electrodes via electrodeposition of polyaniline on nonwoven carbon fiber fabrics
Authors: Leary Jennifer D , Hamouda Farah , Mazé Benoît , Pourdeyhimi Behnam
Published: December 2015

Aniline has been polymerized via electrodeposition onto various nonwoven carbon fiber fabric (CFF) substrates for use as a pseudocapacitive electrochemical capacitor. Four types of CFF were initially tested for double layer capacitance before polyaniline deposition, and again for specific capacitance after deposition. A binder‐free CFF was selected for further analysis due to its high capacitance change following PANI deposition (three orders of magnitude). The aniline monomer concentration, deposition potential, and deposition time were varied and resulting materials were characterized using chrono‐potentiometry, cyclic voltammetry, and scanning electron microscopy. The deposition potential range yielding highest capacitance was found to be between 0.744 and 0.777 V. A solution concentration of 0.5M aniline at a 20 min deposition time resulted in the highest specific capacitance (>80 F/g based on total electrode mass and >300 F/g based on PANI mass) within this study. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43315.
Enhancement of dielectric constants of epoxy thermosets via a fine dispersion of barium titanate nanoparticles
Authors: Li Lei , Zheng Sixun
Published: December 2015

In this study, we examined a facile approach for achieving a fine dispersion of barium titanate (BT) nanoparticles (NPs) in epoxy thermosets. First, the surfaces of BT NPs were modified with poly(ε‐caprolactone) (PCL) via a surface‐initiated ring‐opening polymerization approach. We found that the PCL‐grafted BT NPs were easily dispersed in epoxy thermosets. The fine dispersion of the PCL‐grafted BT NPs in the epoxy thermosets was evidenced by transmission electron microscopy and dynamic mechanical thermal analysis. We found that the organic–inorganic nanocomposites displayed significantly enhanced dielectric constants and low dielectric loss compared to the control epoxy. The nanocomposites containing 14.1 wt % BT NPs possessed dielectric constants as high as at a frequency of 103 Hz. The dielectric loss was measured to be 0.002 at a frequency of 103 Hz. The improved dielectric properties are accounted for the fine dispersion of the BT NPs in the epoxy thermosets. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43322.
Overcoming vocational prejudice: how can skills competitions improve the attractiveness of vocational education and training in the UK?
Authors: Chankseliani Maia , James Relly Susan , Laczik Andrea
Published: December 2015

Raising the attractiveness of vocational education and training (VET) has been on the UK as well as the European agenda for some time, primarily for economic and social development reasons. However, little is known about the role of skills competitions in improving the attractiveness of VET. This study uses data from 110 interviews with WorldSkills competitors and their associates to examine the potential contributions of skills competitions to revitalise VET in the UK. Adopting the enhancement strategy we propose that the experiences of young people who have been internationally recognised for excellence in their respective vocations are inspiring for others. Such experiences have the potential to refocus the attention from a deficit model of VET to the level of excellence that can be achieved through competitions. However, the enhancement strategy in and of itself is not enough to raise the attractiveness of VET. To do so requires consistent policy efforts oriented on spreading excellence throughout the entire VET sector. The ultimate disruption of the policy cycle that perpetuates the existing vocational/academic divide may be achieved through a systemic approach that builds upon the examples of vocational excellence.
Construction of polyketide overproducing Escherichia coli strains via synthetic antisense RNAs based on in silico fluxome analysis and comparative transcriptome analysis
Authors: Meng Hai‐Lin , Xiong Zhi‐Qiang , Song Shu‐Jie , Wang Jianfeng , Wang Yong
Published: December 2015

Rapid assessment and optimization of the incompatible metabolic modules remain a challenge. Here, we developed a systematic approach to characterize the module interactions and improve the problematic modules during the 6‐deoxyerythronolide B (6dEB) biosynthesis in E. coli. Tremendous differences in the overall trends of flux changes of various metabolic modules were firstly uncovered based on in silico fluxome analysis and comparative transcriptome analysis. Potential targets for improving 6dEB biosynthesis were identified through analyzing these discrepancies. All 25 predicted targets at modules of PP pathway and nucleotide metabolism were firstly tested for improving the 6dEB production in E. coli via synthetic antisense RNAs. Down‐regulation of 18 targets genes leads to more than 20% increase in 6dEB yield. Combinatorial repression of targets with greater than 60% increase in 6dEB titer, e.g., anti‐guaB/anti‐zwf led to a 296.2% increase in 6dEB production (210.4 mg/L in flask) compared to the control (53.1 mg/L). This is the highest yield yet reported for polyketide heterologous biosynthesis in E. coli. This study demonstrates a strategy to enhance the yield of heterologous products in the chassis cell and indicates the effectiveness of antisense RNA for use in metabolic engineering.
Exosomal transfer of functional small RNAs mediates cancer‐stroma communication in human endometrium
Authors: Maida Yoshiko , Takakura Masahiro , Nishiuchi Takumi , Yoshimoto Tanihiro , Kyo Satoru
Published: December 2015

Exosomes are small membrane vesicles secreted from a variety of cell types. Recent evidence indicates that human cells communicate with each other by exchanging exosomes. Cancer cells closely interact with neighboring stromal cells, and together they cooperatively promote disease via bidirectional communication. Here, we investigated whether exosomes can play roles in intercellular communication between cancer cells and neighboring fibroblasts. Endometrial fibroblasts were isolated from normal endometrial tissues and from endometrial cancer tissues, and cell‐to‐cell transfer of endometrial cancer cell line Ishikawa‐derived exosomes was examined. The isolated fibroblasts were cultured in conditioned media from CD63‐GFP‐expressing Ishikawa cells, and we found that GFP‐positive exosomes were transferred from Ishikawa cells to the fibroblasts. Next, we introduced a shRNA for a luciferase gene into Ishikawa cells. This shRNA was encapsulated into exosomes, was transferred to the fibroblasts, and then downregulated luciferase expression in the fibroblasts. The mature microRNAs naturally expressed in Ishikawa‐derived exosomes were also transported into the endometrial fibroblasts, and they altered the microRNA expression profiles of the fibroblasts. These results indicated that endometrial cancer cells could transmit small regulatory RNAs to endometrial fibroblasts via exosomes. Our findings document a previously unknown mode of intercellular communication between cancer cells and related fibroblasts in human endometrium.
Natural Mechanism of Origination and Conservation of Monochirality of Amino Acids
Authors: Kadyshevich Elena A. , Ostrovskii Victor E.
Published: December 2015

The proteins belonging to different organisms as well as the natural amino acids as incorporated into proteins occurring free in tissues and lymphatic fluids are, as a rule, levorotatory. This article contains a simple explanation of this structural discrimination and of the extremely slow racemization of natural organic substances. Chirality 28:153–157, 2016. © 2015 Wiley Periodicals, Inc.
The pathological and biochemical identification of possible seed‐lesions of transmitted transthyretin amyloidosis after domino liver transplantation
Authors: Yoshinaga Tsuneaki , Yazaki Masahide , Sekijima Yoshiki , Kametani Fuyuki , Miyashita Kana , Hachiya Naomi , Tanaka Tomohiro , Kokudo Norihiro , Higuchi Keiichi , Ikeda Shu‐ichi
Published: December 2015

The most serious issue in domino liver transplantation (DLT) using liver grafts from patients with transthyretin (TTR)‐related familial amyloid polyneuropathy (FAP) is the development of iatrogenic transmitted amyloidosis (de novo amyloidosis) in DLT‐recipients. However, little is known regarding the mechanisms of the initial stage of amyloid formation in these recipients. We detected initial lesions (possible seed‐lesions) of this iatrogenic amyloidosis in two recipients following liver grafting from FAP patients. Patient 1 underwent DLT at age 65 from an FAP patient with a Val30Met TTR variant and patient 2 received DLT from an FAP patient with a Val30Leu TTR variant at age 32. Patient 2 was started on diflunisal administration from 4 years after DLT. While neither patient had symptoms of FAP, small amyloid deposits were detected on the gastroduodenal mucosae 14 months and 12 years after DLT in patient 1 and patient 2, respectively. The amyloid was analyzed using a laser microdissection system and tandem mass spectrometry. Biochemical analysis indicated that the amyloid was composed mostly of variant TTR produced from the transplanted liver in both patients. In patient 1, wild‐type TTR amyloid was detectable in the duodenal mucosa obtained 2 years after DLT. This is the first study to successfully capture the pathological and biochemical features of initial‐stage amyloid lesions in DLT recipients. The findings clearly indicate that amyloid deposition can start by deposition of variant TTR followed by deposition of wild‐type TTR, and blocking of amyloid seed formation from variant TTR may be a key to prevent or delay the development of DLT‐associated amyloidosis.
Preconcentration of Bismuth(III) by Ultrasound Assisted Emulsification Solidified Floating Organic Drop Microextraction and Analysis by Atomic Absorption Spectrometry
Authors: Tekin Kubilay , Durukan İlknur
Published: December 2015

Ultrasound‐assisted emulsification solidified floating organic drop microextraction was used for preconcentration of trace amounts of bismuth prior to its determination by flame atomic absorption spectrometry. The bismuth ions were extracted by an organic microdrop composed of 1‐dodecanol containing the complexing agent dithizone. The method involves the transferring of the bismuth ion from the aqueous solution to the organic phase under mild conditions. It combines the advantages of simplicity of liquid–liquid microextraction with the quickness of the ultrasound technique. All crucial parameters affecting the analytical performance were studied. Under optimum conditions, the enhancement factor was determined to be 29. The detection limit and precision were obtained as 7.5 ng/mL and 1.03% (n = 9, 100 ng/mL), respectively. The accuracy of the developed method was evaluated by analyzing a certified reference material and was successfully applied for the analysis of water samples.
Darkening on lead‐based pigments: Microbiological contribution
Authors: Rosado Tânia , Gil Milene , Mirão José , Candeias António , Caldeira Ana Teresa
Published: December 2015

The biodeterioration/biodegradation process is an important issue for the conservation of cultural heritage that needs urgent answers to their rehabilitation. In this way, the role of microorganisms in surfaces alteration was exploited. This work revealed a strong relationship between the microbiological proliferation and the damaged areas, evidencing the important role of the microorganisms in mural paintings alteration process. The oxidation of lead‐based pigments noticeably contributes to the pigments alteration, and seems to be correlated with the presence of biodeteriorative microorganisms. The study of the mechanisms underlying the microbiological attack of mural paintings has been explored to understand as much as possible the proliferative ability and biodeteriorative capacity of the microorganisms, related to darkening on lead‐based pigments. © 2015 Wiley Periodicals, Inc. Col Res Appl, 41, 294–298, 2016
Epitope located N‐glycans impair the MHC‐I epitope generation and presentation
Authors: Chiritoiu Gabriela N. , Jandus Camilla , Munteanu Cristian V.A. , Ghenea Simona , Gannon Philippe O. , Romero Pedro , Petrescu Stefana M.
Published: December 2015

The degradation process of the antigens specific to MHC‐I presentation depends mainly on the proteasomal proteases in the cytosol. However, since many antigens are glycoproteins, including tumor antigens or viruses envelope proteins, their glycosylation status could also affect their processing and presentation. Here, we investigate the processing of tyrosinase, a multiple glycosylated tumor antigen overexpressed in human malignant melanoma. By LC‐MS/MS analysis of human tyrosinase expressed in a melanoma cell, we show that all seven sites of tyrosinase are at least partially N‐glycosylated. Using human CD8+ T‐cell clones specific for the tyrosinase epitope YMDGTMSQV (369–377), including an N‐glycosylation site, we found that transfectants of single and triple N‐glycosylation mutants are recognized by specific T cells. Importantly, single, triple, and the aglycosylated tyrosinase mutants lacking the epitope located N‐glycosylation site (N371D) were able to trigger higher CD8+ T‐cell activation. The LC/MS analysis showed significant increase of the amount of YMDGTMSQV peptide resulted from accelerated oligomerization and degradation of aglycosylated mutants. The generation of the antigenic peptide by the antigen processing machinery is therefore largely independent of tyrosinase N‐glycosylation. However, while distal N‐glycans had no effect on the epitope generation, the mutants lacking the N371 glycan generated the antigenic peptide more efficiently. We conclude that epitope located N‐glycans limit the ability of human tyrosinase to provide HLA‐A2‐restricted antigen for recognition by specific CD8+ T cells.
Role of perioperative antibiotic treatment in parotid gland surgery
Authors: Shkedy Yotam , Alkan Uri , Roman Benjamin R. , Hilly Ohad , Feinmesser Raphael , Bachar Gideon , Mizrachi Aviram
Published: December 2015

Background

The value of routine prophylactic antibiotic treatment in parotid gland surgery remains undetermined.

Methods

A retrospective analysis was conducted of all patients who underwent parotidectomy at a university‐affiliated tertiary care center between 1992 and 2009. Patients with insufficient data, specifically regarding postoperative complications and antibiotic administration were excluded from the study cohort.

Results

A total of 593 patients underwent parotidectomy during the study period. After exclusion, 464 patients were eligible for the study. Perioperative antibiotic treatment was given to 206 patients (45%). There was no difference in wound infection rates between patients who received perioperative antibiotic therapy and those who did not (p = .168). Multivariate analysis showed that female sex, neck dissection, and drain output >50 cc/24 hours were predictive of postoperative wound infection.

Conclusion

Routine prophylactic antibiotic treatment has no role in parotid gland surgery. Perioperative antibiotic treatment is recommended for patients undergoing extensive parotid gland surgery with neck dissection. © 2015 Wiley Periodicals, Inc. Head Neck 38: E1876–E1880, 2016
Genetic variation in STAT4 predicts response to interferon‐α therapy for hepatitis B e antigen‐positive chronic hepatitis B
Authors: Jiang De‐Ke , Wu Xiaopan , Qian Ji , Ma Xiao‐Pin , Yang Jingmin , Li Zhuo , Wang Runhua , Sun Li , Liu Fang , Zhang Pengyin , Zhu Xilin , Wu Jia , Chen Kangmei , Conran Carly , Zheng S. Lilly , Lu Daru , Yu Long , Liu Ying , Xu Jianfeng
Published: December 2015

Interferon (IFN)‐α is a first‐line therapy for chronic hepatitis B (CHB) patients but only initiates a response in a minority of patients. A genetic variant, rs7574865 in STAT4, was recently reported to be associated with risk of developing CHB and hepatitis B virus‐related hepatocellular carcinoma. We aimed to determine whether this variant is associated with the response to IFNα treatment for hepatitis B e antigen (HBeAg)‐positive CHB patients. We studied 466 HBeAg‐positive CHB patients who received either IFNα‐2b (n = 224) or pegylated IFNα‐2a (n = 242) therapy for 48 weeks and were followed for an additional 24 weeks. The rate of sustained virologic response (SVR), defined as HBeAg seroconversion along with hepatitis B virus DNA level <2000 copies/mL at week 72, was compared among patients with different genotypes of rs7574865. After 48 weeks of treatment and 24 weeks off treatment, the SVR rates in the IFNα‐2b and pegylated IFNα‐2a therapy groups were 30.4% and 28.9%, respectively. Compared to the rs7574865 GT/TT genotype, the GG genotype (a risk factor of CHB and hepatitis B virus‐related hepatocellular carcinoma) was significantly associated with a reduced SVR rate in both patients who received IFNα‐2b therapy (21.1% versus 37.2%, P = 0.01) and those who received pegylated IFNα‐2a therapy (18.0% versus 41.2%, P = 9.74 × 10‐5). In joint analysis of the 466 patients, the GG genotype was associated with an approximately half SVR rate compared to the GT/TT genotype (19.3% versus 39.1%, P = 4.15 × 10‐6). A multivariate logistic regression model including rs7574865 and clinical variables showed that rs7574865 was the most significant factor for the prediction of SVR. Conclusion: STAT4 rs7574865 is a reliable predictor of response to IFNα therapy for HBeAg‐positive CHB patients and may be used for optimizing the treatment of CHB. (Hepatology 2016;63:1102–1111)
Safety and efficacy of ledipasvir/sofosbuvir for the treatment of genotype 1 hepatitis C in subjects aged 65 years or older
Authors: Saab Sammy , Park Sarah H. , Mizokami Masashi , Omata Masao , Mangia Alessandra , Eggleton Ed , Zhu Yanni , Knox Steven J. , Pang Phil , Subramanian Mani , Kowdley Kris , Afdhal Nezam H.
Published: December 2015

Elderly subjects have been historically underrepresented in clinical trials involving antiviral hepatitis C therapies. The aim of this analysis was to retrospectively evaluate the safety and efficacy of ledipasvir/sofosbuvir (LDV/SOF) by age groups of <65 years versus ≥65 years among subjects enrolled in phase 3 trials. Four open‐label phase 3 clinical trials evaluated the safety and efficacy of LDV/SOF with or without ribavirin (RBV) for the treatment of genotype 1 chronic hepatitis C virus. Sustained virological response at 12 weeks, treatment‐emergent adverse events (AEs), and graded laboratory abnormalities were analyzed according to age group. Of the 2293 subjects enrolled in four phase 3 trials, 264 (12%) were ≥65 years of age, of whom 24 were aged ≥75 years. Sustained virological response at 12 weeks was achieved by 97% (1965/2029) of subjects aged <65 years and 98% (258/264) of subjects aged ≥65 years. The most common AEs in both LDV/SOF groups that occurred in ≥10% of subjects were headache and fatigue. The rate of study discontinuation due to AEs was similar in the two age cohorts. The use of RBV in 1042 (45%) subjects increased the number of AEs, treatment‐related AEs, and AEs leading to study drug modification/interruption, particularly among elderly subjects. Conclusions: LDV/SOF with or without RBV was highly effective for treatment of genotype 1 chronic hepatitis C virusin subjects aged 65 and older. Addition of RBV did not increase sustained virological response at 12 weeks rates but led to higher rates of AEs, especially in elderly subjects. (Hepatology 2016;63:1112–1119)
Recruitment of mesenchymal stem cells and macrophages by dual release of stromal cell‐derived factor‐1 and a macrophage recruitment agent enhances wound closure
Authors: Kim Yang‐Hee , Tabata Yasuhiko
Published: December 2015

In this study, the wound closure of mouse skin defects was examined in terms of recruitment of mesenchymal stem cells (MSC) and macrophages. For the cells recruitment, stromal derived factor‐1 (SDF‐1) of a MSC recruitment agent and sphingosine‐1 phosphate agonist (SEW2871) of a macrophages recruitment agent were incorporated into gelatin hydrogels, and then released in a controlled fashion. When applied to a skin wound defect of mice, gelatin hydrogels incorporating mixed 500 ng SDF‐1 and 0.4, 0.8, or 1.6 mg SEW2871‐micelles recruited a higher number of both MSC and macrophages than those incorporating SDF‐1 or phosphate buffered saline. However, the number of M1 phenotype macrophages for the hydrogel incorporating mixed SDF‐1 and SEW2871‐micelles recruited was remarkably low to a significant extent compared with that for those hydrogel incorporating 0.4, 0.8, or 1.6 mg SEW2871‐micelles. On the other hand, the number of M2 macrophages 3 days after the implantation of the hydrogels incorporating SDF‐1 and 0.4 mg SEW2871‐micelles significantly increased compared with that for other hydrogels. In vivo experiments revealed the hydrogels incorporating SDF‐1 and 0.4 mg SEW2871‐micelles promoted the wound closure of skin defect to a significant stronger extent than those incorporating SEW2871‐micelles, SDF‐1, and a mixture of SDF‐1 and higher doses of SEW2871‐micelles. It is concluded that the in vivo recruitment of MSC and macrophages to the defects may contribute to the tissue regeneration of skin wound. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 942–956, 2016.
Validation of eGFP fluorescence intensity for testing in vitro cytotoxicity according to ISO 10993‐5
Authors: Miller Felicitas , Hinze Ulf , Chichkov Boris , Leibold Wolfgang , Lenarz Thomas , Paasche Gerrit
Published: December 2015

ISO 10993‐5 provides one of the accepted standards for testing the biotoxicity of new materials. All of the recommended test procedures rely upon the uptake or metabolism of dye by living cells. Results of direct contact tests can be potentially compromised by interaction or adsorption of the dye or its metabolic products. Therefore, the aim of the current study was to validate the use of the eGFP signal of transfected NIH‐3T3 fibroblasts with the results of the MTT test in order to provide a test procedure that is very close to the ISO 10993‐5 but has the advantage of not relying on the addition of dye. Our tests show that the MTT assay detects cytotoxicity in the eGFP NIH‐3T3 cells at least as well as in the L929 cells. To facilitate the validation, we chose to integrate the fluorescence measurements into the MTT test procedure. To that end, an additional washing step was introduced. Additionally, medium without phenol red was used, resulting in a very high correlation of both measurements. Without these modifications, the fluorescence test was comparable to the MTT test in its ability to detect the cytotoxic potential of substances; however, it did result in slightly elevated IC50 concentrations. As the results of both tests correlated highly, measurement of the eGFP signal appears to present a reliable tool for detecting cytotoxicity of materials in line with the ISO 10993‐5 norm with the advantage of avoiding the addition of dyes and the subsequent potential interaction with test materials. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 715–722, 2017.
Fabrication of chitosan/gallic acid 3D microporous scaffold for tissue engineering applications
Authors: Thangavel Ponrasu , Ramachandran Balaji , Muthuvijayan Vignesh
Published: December 2015

This study explores the potential of gallic acid incorporated chitosan (CS/GA) 3D scaffolds for tissue engineering applications. Scaffolds were prepared by freezing and lyophilization technique and characterized. FTIR spectra confirmed the presence of GA in chitosan (CS) gel. DSC and TGA analysis revealed that the structure of chitosan was not altered due to the incorporation of GA, but thermal stability was significantly increased compared to the CS scaffold. SEM micrographs showed smooth, homogeneous, and microporous architecture of the scaffolds with good interconnectivity. CS/GA scaffolds exhibited approximately 90% porosity on average, increased swelling (600–900%) and controlled biodegradation (15‐40%) in PBS (pH 7.4 at 37°C) with 1 mg/mL of lysozyme. CS/GA scaffolds showed 2‐4 fold decrease in CFUs (p < 0.05) for both gram positive and gram negative bacteria compared to the CS scaffold. Cytotoxicity of these scaffolds was evaluated using NIH 3T3 L1 fibroblast cells. CS/GA 0.25% scaffold showed similar viability with CS scaffold at 24 and 48 h. CS/GA scaffolds (0.5–1.0%) showed 60–75% viability at 24 h and 90% at 48 h. SEM images showed that an increased cell attachment was observed for CS/GA scaffolds compared to CS scaffolds. These findings authenticate that CS/GA scaffolds were cytocompatible and would be useful for tissue engineering applications. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 750–760, 2016.