bluuebunny/crossref_metadata_2025 huggingface dataset:
Serum hepatitis B core‐related antigen as a treatment predictor of pegylated interferon in patients with HBeAg‐positive chronic hepatitis B
Authors: Chuaypen Natthaya , Posuwan Nawarat , Payungporn Sunchai , Tanaka Yasuhito , Shinkai Noboru , Poovorawan Yong , Tangkijvanich Pisit
Published: December 2015

Background & Aims

The role of quantitative serum hepatitis B core‐related antigen (HBcrAg) in patients with chronic hepatitis B (CHB) receiving pegylated interferon (PEG‐IFN) is unclear. This study was aimed at comparing its usefulness with quantitative HBsAg in patients with HBeAg‐positive CHB receiving PEG‐IFN therapy.

Methods

A total 46 patients treated with PEG‐IFN for 48 weeks were retrospectively analysed. Intrahepatic covalently closed circular DNA (cccDNA) from paired liver biopsies and serial serum HBsAg and HBcrAg during therapy were assessed.

Results

Virological response (VR), defined as HBeAg clearance and HBV DNA <2000 IU/ml at 24 weeks post treatment, was achieved in 15 (32.6%) patients. Responders had significantly higher cccDNA decline from baseline compared with non‐responders. Baseline HBsAg and HBcrAg were correlated with cccDNA (r = 0.424, P = 0.020 and r = 0.564, P = 0.001, respectively), and changes in the corresponding markers during therapy were correlated with cccDNA reduction (r = 0.579, P = 0.001 and r = 0.503, P = 0.005, respectively). Responders showed more rapid decline of both markers during therapy compared with non‐responders. In multivariate analysis, serum HBcrAg at week 12 was identified as a predictor of VR. The optimal cut‐off value for HBcrAg (log10 8.0 U/ml) provided negative predictive value (NPV) of achieving VR at weeks 12 and 24 of 94.4 and 100%, respectively, while using HBsAg > 20 000 IU/ml provided NPV of 80 and 100% respectively.

Conclusions

The convenient quantitative HBcrAg represented a reliable marker of intrahepatic cccDNA. Monitoring HBcrAg levels during PEG‐IFN therapy may help identify patients with a very low probability of response comparable to, if not better than, quantitative HBsAg.
The school environment and adolescent physical activity and sedentary behaviour: a mixed‐studies systematic review
Authors: Morton K. L. , Atkin A. J. , Corder K. , Suhrcke M. , van Sluijs E. M. F.
Published: December 2015

There is increasing academic and policy interest in interventions aiming to promote young people's health by ensuring that the school environment supports healthy behaviours. The purpose of this review was to summarize the current evidence on school‐based policy, physical and social‐environmental influences on adolescent physical activity and sedentary behaviour. Electronic databases were searched to identify studies that (1) involved healthy adolescents (11–18 years old), (2) investigated school‐environmental influences and (3) reported a physical activity and/or sedentary behaviour outcome or theme. Findings were synthesized using a non‐quantitative synthesis and thematic analysis. Ninety‐three papers of mixed methodological quality were included. A range of school‐based policy (e.g. break time length), physical (e.g. facilities) and social‐environmental (e.g. teacher behaviours) factors were associated with adolescent physical activity, with limited research on sedentary behaviour. The mixed‐studies synthesis revealed the importance of specific activity settings (type and location) and intramural sport opportunities for all students. Important physical education‐related factors were a mastery‐oriented motivational climate and autonomy supportive teaching behaviours. Qualitative evidence highlighted the influence of the wider school climate and shed light on complexities of the associations observed in the quantitative literature. This review identifies future research needs and discusses potential intervention approaches to be considered.
A national quality registers as a tool to audit items of the fundamentals of care to older patients with hip fractures
Authors: Hommel Ami , Bååth Carina
Published: December 2015

Background

The Swedish healthcare system has a unique resource in the national quality registers. A national quality registry contains individualised data concerning patient problems, medical interventions and outcomes after treatment, within all healthcare settings. Many healthcare settings face challenges related to the way they deliver the fundamentals of care, therefore, it is important to audit the outcome. It is estimated that the number of people aged 80 years or older will have almost quadrupled between 2000 and 2050. Hip fracture has been recognised as the most serious consequence of osteoporosis because of the risk of its complications, which include pain, acute confusional state, pressure ulcers, infections, disability, diminished quality of life and mortality.

The aim of this study was therefore to explore if and how a national quality register can be used as an audit tool for the fundamentals of care when it concerns older patients suffering from a hip fracture.

Method

For this study we retrospectively selected and audited variables retrieved from the national quality hip fracture register. The audit included 1083 patients 80 years and older, consecutively admitted to a university hospital in the south of Sweden, in 2011–2013.

Results

Nearly half of the patients were admitted from their own homes and were living alone. Almost half of the patients could walk outdoors before the fracture occurred. After 4 months, 28.5% of the patients walked outdoors. Additionally, after 4 months about 30% of the patients were still suffering from pain after hip fracture surgery and still using analgesics. There was a reduction in length of stay between 2011 and 2013. As a part of the national quality register the questions from EQ5D were used before surgery and after 4 months. Before discharge from hospital there were less registered complications in 2012 and 2013 compared with 2011.

Conclusions

The national hip fracture quality register allows healthcare staff to analyse nursing outcomes and to highlight some fundamental aspects of care.

Implications for practice

Greater awareness, among hospital staff, of risk factors for complications in hip fracture patients may lead to improved patient care. Through registration in a quality register and working with the results we as Registered Nurses can ensure quality health care for older adults.
Diagnosis and clinical presentation of hemoglobin Kirksey
Authors: Hitaka Daisuke , Arai Junichi
Published: December 2015

More than 1000 hemoglobin (Hb) variants have been identified. Hb Kirksey, a rare Hb variant involving mutation at codon 94 of α2‐globin, is associated with low oxygen affinity. To our knowledge, there is no case report on Hb Kirksey in the literature, and, accordingly, the clinical features of patients with Hb Kirksey are currently unknown. We report here the case of a neonate who presented with asymptomatic low oxygen saturation on pulse oximetry (SpO2) just after birth, together with clinical analysis of the sister and cousin. Based on the presentations, Hb Kirksey does not seem to be associated with clinical abnormalities. Discrepancy between SpO2 and partial pressure of arterial oxygen (PaO2) detected on arterial blood gas analysis provided a clue to the diagnosis. Hb variants should be suspected and arterial blood gases should be measured in pediatric cases of unexplained low SpO2.
Sex moderates the association between symptoms of anxiety, but not obsessive compulsive disorder, and error‐monitoring brain activity: A meta‐analytic review
Authors: Moser Jason S. , Moran Tim P. , Kneip Chelsea , Schroder Hans S. , Larson Michael J.
Published: December 2015

Sex differences in cognition and emotion are particularly active areas of research. Much of this work, however, focuses on mean‐level differences between the sexes on cognitive and/or emotional variables in isolation. In this article, we are primarily concerned with how sex affects associations between cognition and emotion, or cognition‐emotion interactions. Specifically, the purpose of this paper is to shed light on a gap in our understanding of how sex affects the relationship between error monitoring, a core component of cognitive control, and anxiety. Using meta‐analysis, we show that the relationship between symptoms of anxiety and a neurophysiological marker of error monitoring—the error‐related negativity (ERN)—is significantly greater in women than men such that women, but not men, with higher levels of anxiety show a larger ERN. This sex difference held true across studies of anxiety‐specific symptoms but not studies of obsessive‐compulsive symptoms. These findings underscore the need to consider sex in studies of anxiety and the ERN as well as support growing evidence indicating that obsessive‐compulsive problems are distinguishable from other anxiety‐specific problems across multiple levels of analysis. Overall, we conclude that ignoring sex in studies of cognition‐emotion interactions is unacceptable. Rather, future research that continues to tackle questions related to sex differences in associations between cognition and emotion will more likely lead to advancements in basic and applied sciences of relevance to health and human behavior.
Increasing awareness of sex differences in airway diseases
Authors: Raghavan Deepa , Jain Raksha
Published: December 2015

There is growing epidemiologic data demonstrating sex differences with respect to prevalence and progression of airway diseases, including asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF) and non‐CF‐related bronchiectasis. In asthma, for example, young boys have increased exacerbations and higher morbidity than girls which distinctly reverses after adolescence and into adulthood. In COPD, a disease that was historically considered an illness of men, the number of women dying per year is now greater than in men. Finally, women with CF‐related bronchiectasis have a decreased median life expectancy relative to men and a higher risk of respiratory infections despite equal prevalence of the disease. A number of studies now exist demonstrating mechanisms behind these sex differences, including influences of genetic predisposition, sex hormones and comorbidities. The notable sex disparity has potential diagnostic, therapeutic and prognostic implications and for the practicing respiratory or general physician, a familiarity with these distinctions may augment effective management of patients with airway diseases. This review seeks to concisely summarize the data regarding gender‐based differences in airway diseases, outline the current understanding of contributing factors and discuss therapeutic implications for clinicians.
The Phenotypic and Functional Features of Human M2 Macrophages Generated Under Low Serum Conditions
Authors: Sakhno L. V. , Shevela E. Ya. , Tikhonova M. A. , Ostanin A. A. , Chernykh E. R.
Published: December 2015

The phenotypic and functional features of human M2 macrophages, in particular, their immunosuppressive activity, can considerably vary depending on M2 polarizing stimulus. This study was aimed at the investigation of cytokine production and pro‐apoptogenic/inhibitory molecule expression in macrophages generated with GM‐CSF using either standard conditions (M1) or deficiency of serum/growth factors (M2‐LS cells). In contrast to M1, M2‐LS cells were characterized by an enhanced content of CD206+, B7‐H1+, FasL+ and TRAIL+ cells along with a decreased production of IFN‐γ, IL‐5, IL‐6, IL‐13, TNF‐α, IL‐17 and MCP‐1. In addition, M2‐LS exhibited a lower T cell stimulatory activity in MLC that was associated with the higher numbers of apoptotic and the lower numbers of proliferating T cells. B7‐H1 plays a key role in M2‐LS‐mediated cytotoxic effects as the neutralization of B7‐H1 reduces the apoptosis‐inducing activity of M2‐LS, while the blocking of CD206 and TRAIL reduces the cytostatic activity of M2 macrophages.
The Active Metabolite of Vitamin D3 as a Potential Immunomodulator
Authors: Szymczak I. , Pawliczak R.
Published: December 2015

In the past, vitamin D was known for its classical, skeletal action as a regulator of calcium and bone homoeostasis. Currently, vitamin D was found to have a role in numerous physiological processes in the human body; thus, vitamin D has pleiotropic activity. The studies carried out in the past two decades showed the role of vitamin D in the regulation of immune system functions. Basically, these effects may be mediated not only via endocrine mechanism of circulating calcitriol but also via paracrine one (based on cell–cell communication that leads to production of signal inducing the changes in nearby/adjacent cells and modulating their differentiation or behaviour) and intracrine mechanism (the action of vitamin D inside a cell) of 1,25‐dihydroxycholecalciferol (1,25(OH)2D3) synthetized from its precursor 25‐hydroxyvitamin D3 (25(OH)D3). Both vitamin D receptor (VDR) and 25‐hydroxyvitamin D3 1‐α‐hydroxylase (CYP27B1) are expressed in several types of immune cells (i.e. antigen presenting cells, T and B cells), and thus, they are able to synthetize the bioactive form of vitamin D that modulates both the innate and adaptive immune system. This review discusses the role of vitamin D as regulator of immune system, and our understanding of how vitamin D regulates both adaptive and innate immunity as well as inflammatory cascade on the cellular level.
Kainate receptor pore‐forming and auxiliary subunits regulate channel block by a novel mechanism
Authors: Brown Patricia M. G. E. , Aurousseau Mark R. P. , Musgaard Maria , Biggin Philip C. , Bowie Derek
Published: December 2015

Key points

Kainate receptor heteromerization and auxiliary subunits, Neto1 and Neto2, attenuate polyamine ion‐channel block by facilitating blocker permeation.

Relief of polyamine block in GluK2/GluK5 heteromers results from a key proline residue that produces architectural changes in the channel pore α‐helical region.

Auxiliary subunits exert an additive effect to heteromerization, and thus relief of polyamine block is due to a different mechanism.

Our findings have broad implications for work on polyamine block of other cation‐selective ion channels.

Channel block and permeation by cytoplasmic polyamines is a common feature of many cation‐selective ion channels. Although the channel block mechanism has been studied extensively, polyamine permeation has been considered less significant as it occurs at extreme positive membrane potentials. Here, we show that kainate receptor (KAR) heteromerization and association with auxiliary proteins, Neto1 and Neto2, attenuate polyamine block by enhancing blocker permeation. Consequently, polyamine permeation and unblock occur at more negative and physiologically relevant membrane potentials. In GluK2/GluK5 heteromers, enhanced permeation is due to a single proline residue in GluK5 that alters the dynamics of the α‐helical region of the selectivity filter. The effect of auxiliary proteins is additive, and therefore the structural basis of polyamine permeation and unblock is through a different mechanism. As native receptors are thought to assemble as heteromers in complex with auxiliary proteins, our data identify an unappreciated impact of polyamine permeation in shaping the signalling properties of neuronal KARs and point to a structural mechanism that may be shared amongst other cation‐selective ion channels.
Signaling Responses After Varying Sequencing of Strength and Endurance Training in a Fed State
Authors: Jones Thomas W. , Walshe Ian H. , Hamilton David L. , Howatson Glyn , Russell Mark , Price Oliver J. , Gibson Alan St Clair , French Duncan N.
Published: December 2015

Purpose:

To compare anabolic signaling responses to differing sequences of concurrent strength and endurance training in a fed state.

Methods:

Eighteen resistance-trained men were randomly assigned to the following experimental conditions: strength training (ST), strength followed by endurance training (ST-END), or endurance followed by strength training (END-ST). Muscle tissue samples were taken from the vastus lateralis before each exercise protocol, on cessation of exercise, and 1 h after cessation of strength training. Tissue was analyzed for total and phosphorylated (p-) signaling proteins linked to the mTOR and AMPK networks.

Results:

Strength-training performance was similar between ST, ST-END, and END-ST. p-S6k1 was elevated from baseline 1 h posttraining in ST and ST-END (both P < .05). p-4E-BP1 was significantly lower than baseline post-ST (P = .01), whereas at 1 h postexercise in the ST-END condition p-4E-BP1 was significantly greater than postexercise (P = .04). p-ACC was elevated from baseline both postexercise and 1 h postexercise (both P < .05) in the END-ST condition. AMPK, mTOR, p38, PKB, and eEF2 responded similarly to ST, ST-END, and END-ST. Signaling responses to ST, ST-END, and END were largely similar. As such it cannot be ascertained which sequence of concurrent strength and endurance training is most favorable in promoting anabolic signaling.

Conclusions:

In the case of the current study an acute bout of concurrent training of differing sequences elicited similar responses of the AMPK and mTOR networks.
Whole-Genome Sequences of Novel Porcine Circovirus Type 2 Viruses Detected in Swine from Mexico and the United States
Authors: Harmon Karen M. , Gauger Phillip C. , Zhang Jianqiang , Piñeyro Pablo E. , Dunn Derek D. , Chriswell Amy J.
Published: December 2015

A unique porcine circovirus type 2 capsid protein (ORF2) sequence was detected in swine samples submitted to the Iowa State University Veterinary Diagnostic Laboratory. The complete genome sequences of four viruses, one from Mexico and three from the United States, were determined to further characterize this novel PCV2 genotype.
Draft Genome Sequence of
Pedobacter
sp. Strain Hv1, an Isolate from Medicinal Leech Mucosal Castings
Authors: Ott Brittany M. , Beka Lidia , Graf Joerg , Rio Rita V. M.
Published: December 2015

The
Pedobacter
sp. Hv1 strain was isolated from the medicinal leech,
Hirudo verbana,
mucosal castings. These mucosal sheds have been demonstrated to play a role in horizontal symbiont transmission. Here, we report the draft 4.9 Mbp genome sequence of
Pedobacter
sp. strain Hv1.
Genome Sequence of Bradyrhizobium tropiciagri Strain CNPSo 1112
T
, Isolated from a Root Nodule of
Neonotonia wightii
Authors: Delamuta Jakeline Renata Marçon , Gomes Douglas Fabiano , Ribeiro Renan Augusto , Chueire Ligia Maria Oliveira , Souza Renata Carolini , Almeida Luiz Gonzaga Paula , Vasconcelos Ana Tereza Ribeiro , Hungria Mariangela
Published: December 2015

CNPSo 1112
T
is a nitrogen-fixing symbiont of perennial soybean, a tropical legume forage. Its draft genome indicates a large genome with a circular chromosome and 9,554 coding sequences (CDSs). Operons of nodulation, nitrogen fixation, and uptake hydrogenase were present in the symbiotic island, and the genome encompasses several CDSs of stress tolerance.
Draft Genome Sequence of
Caloranaerobacter
sp. TR13, an Anaerobic Thermophilic Bacterium Isolated from a Deep-Sea Hydrothermal Vent
Authors: Zhou Meixian , Xie Yunbiao , Dong Binbin , Liu Qing , Chen Xiaoyao
Published: December 2015

Here, we report the draft 2,261,881-bp genome sequence of
Caloranaerobacter
sp. TR13, isolated from a deep-sea hydrothermal vent on the East Pacific Rise. The sequence will be helpful for understanding the genetic and metabolic features, as well as potential biotechnological application in the genus
Caloranaerobacter
.
Simulation of the experimental thermal cleaning procedure in LiF:Mg,Ti and investigation of the origin of the first-order kinetics of its peaks
Authors: Halimi Mokhtar , Kadri Dahane , Mokeddem Abdelmalek
Published: December 2015

In this paper, the simulation of the thermal cleaning procedure in LiF:Mg,[Formula: see text]Ti as well as the investigation of the origin of its glow-peaks have been performed by six electron trapping levels and one recombination center phenomenological model. The most generally accepted parameter of the glow-peaks 1 to 5 of LiF:Mg,[Formula: see text]Ti is their first-order kinetics; this was particularly investigated by the use of the random variation of the kinetic parameters method.
Stringy Maxwell charge and the magnetic dipole moment
Authors: Hammond Richard T.
Published: December 2015

It is shown that stringy charge leads naturally to an observable magnetic moment of the electron.
New determination of 12C(α,γ)16O reaction rate
Authors: Oulebsir N.
Published: December 2015

The reaction [Formula: see text]C([Formula: see text])[Formula: see text]O was investigated through the direct [Formula: see text]-transfer reaction (7Li,t) at 28 and 34 MeV incident energies. We determined the reduced [Formula: see text]-widths of the sub-threshold 2[Formula: see text] and 1[Formula: see text] states of [Formula: see text]O from the DWBA analysis of the transfer reaction [Formula: see text]C(7Li,t)[Formula: see text]O performed at two incident energies. The obtained result for the 2[Formula: see text] and 1[Formula: see text] sub-threshold resonances as introduced in the [Formula: see text]-matrix fitting of radiative capture and elastic-scattering data to determine the E2 and E1 [Formula: see text]-factor from 0.01[Formula: see text]MeV to 4.2[Formula: see text]MeV in the center-of-mass energy. After determining the astrophysic factor of [Formula: see text]C([Formula: see text])[Formula: see text]O S(E) with Pierre Descouvement code, I determined numerically the new reaction rate of this reaction at a different stellar temperature (0.06 Gk-2 GK). The [Formula: see text]C([Formula: see text])[Formula: see text]O reaction rate at T[Formula: see text] is [7.21[Formula: see text]] × 10[Formula: see text] cm3 s[Formula: see text] mol[Formula: see text]. Some comparisons and discussions about our new [Formula: see text]C([Formula: see text])[Formula: see text]O reaction rate are presented. The agreements of the reaction rate below T[Formula: see text] between our results and with those proposed by NACRE indicate that our results are reliable, and they could be included in the astrophysical reaction rate network.
Moment problems in an infinite number of variables
Authors: Alpay Daniel , Jorgensen Palle E. T. , Kimsey David P.
Published: December 2015

Let [Formula: see text]. Given a closed set [Formula: see text] and [Formula: see text], where [Formula: see text] denotes the set of tuples of nonnegative integers [Formula: see text] with [Formula: see text] for finitely many [Formula: see text], the [Formula: see text]-moment problem on [Formula: see text] entails determining whether or not there exists a measure [Formula: see text] on [Formula: see text] so that [Formula: see text] and [Formula: see text] We prove that [Formula: see text] exists if and only if a natural analogue of the Riesz–Haviland functional [Formula: see text] is [Formula: see text]-positive, i.e. if [Formula: see text] is any polynomial which is nonnegative for all [Formula: see text], then [Formula: see text] We will also provide a sufficient condition for [Formula: see text] to be unique, an analogue of a celebrated theorem of K. Schmüdgen and an application to stochastic processes.
SET UP FOR GROWTH? — AN EXPLORATORY ANALYSIS OF THE RELATIONSHIP OF GROWTH INTENTION AND BUSINESS MODELS
Authors: MAHDJOUR SARAH
Published: December 2015

What do growth-oriented business models look like? While several economic theories, such as the theory of the firm, are based on the assumption that firms aim to maximise their profits, past research has shown that growth intention is heterogeneous among firms and that many business owners prefer to keep their firm at a size that they can manage with few resources. This paper explores the relationship of growth intention and business models, based on a sample of 135 German ICT businesses. Following an exploratory approach, Mann–Whitney U tests are applied to analyse how different business model designs correspond with different levels of growth intention. The results indicate that growth intention relates to business owners’ decisions regarding the provision of consulting services, the level of standardisation in offered products and services, the choice of addressed markets, the implementation of competitive strategies based on cost efficiency and of revenue streams based on one-time- and performance-based payments. Furthermore, the results show that growth oriented firms are no more likely than non-growth oriented firms to adapt their business models dynamically to changed internal or external conditions.
IS RADICAL INNOVATION MANAGEMENT MISUNDERSTOOD? PROBLEMATISING THE RADICAL INNOVATION DISCIPLINE
Authors: KRISTIANSEN JIMMI NORMANN , GERTSEN FRANK
Published: December 2015

This paper poses a critical view on radical innovation (RI) management research and practice. The study investigates how expected RI performance influences firms’ understanding of their RI capability. RI performance is often based on output measures such as market shares or fiscal return. On the contrary, RI capability building advocates for exploration, learning, and accepting uncertainty. Hence, RI capability building often focuses on the processes of the firms, and not the outcome. Thus, it is argued that the RI capability-building and RI performance expectations are based on different managerial orientations. Coupling a discussion of the literature with case findings from four large international firms, this paper identifies a discrepancy between RI capability-building and RI performance within literature and practice. This is regarded to be a major contributing factor to RI program failure. This study presents three misunderstandings related to RI research and practice and discusses implications of these findings.
CRISIS-DRIVEN INNOVATION: THE CASE OF HUMANITARIAN INNOVATION
Authors: BESSANT JOHN , RUSH HOWARD , TRIFILOVA ANNA
Published: December 2015

Innovation is often presented as a survival issue and this provides a powerful metaphor to focus attention on the need to manage it effectively. But in the humanitarian context it takes on a very literal meaning. Crises, whether natural or man-made, require rapid problem solving if agencies and aid workers are to avoid the huge negative impacts of such disasters.

That makes consideration of how innovation takes place in this sector an urgent challenge. How can the humanitarian sector best organise to enable innovation and what are the roles for key actors — donors, agencies, and most importantly ‘users’? Our paper summarises the nature of the challenge and reviews experience so far in humanitarian innovation (HI).

There is a second issue which we also explore. Arguably crisis conditions provide a “laboratory” for exploring alternative approaches and generating novel innovation trajectories which might diffuse more widely — the concept of “reverse innovation.” Are there lessons which could be learned for mainstream innovation management? And if so, what are the mechanisms which might enable such learning and experience exchange to take place?
INNOVATION AND GROWTH IN THE CITY REGION: MICROECONOMIC EVIDENCE OF ASYMMETRIES
Authors: DEVINS DAVID , LODORFOS GEORGE , KOSTOPOULOS IOANNIS , WEBBER DON
Published: December 2015

Using a micro-economic perspective, this study explores the type, scale and impact of innovation activities that small and medium-sized enterprises (SMEs) undertake with focus on the sub-national level. Based on a representative sample of 250 independent SMEs, based in the Leeds City Region (LCR) and using quantitative methods, this study provides evidence of the types of innovation that are most likely to result in achieving policy aims associated with for example the development of international markets or growth of employment opportunities. The findings show that there is a need for aligning business support for innovation with human capital (education/training) interventions. Furthermore, the findings provide a strong evidence base, at the local level, to inform the development of innovation policy and illustrate the connections between different types of innovation SMEs’ growth at regional level.
An organotypic slice model for ex vivo study of neural, immune, and microbial interactions of mouse intestine
Authors: Schwerdtfeger Luke A. , Ryan Elizabeth P. , Tobet Stuart A.
Published: December 2015

Organotypic tissue slices provide seminatural, three-dimensional microenvironments for use in ex vivo study of specific organs and have advanced investigative capabilities compared with isolated cell cultures. Several characteristics of the gastrointestinal tract have made in vitro models for studying the intestine challenging, such as maintaining the intricate structure of microvilli, the intrinsic enteric nervous system, Peyer's patches, the microbiome, and the active contraction of gut muscles. In the present study, an organotypic intestinal slice model was developed that allows for functional investigation across regions of the intestine. Intestinal tissue slices were maintained ex vivo for several days in a physiologically relevant environment that preserved normal enterocyte structure, intact and proliferating crypt cells, submucosal organization, and muscle wall composure. Cell death was measured by a membrane-impermeable DNA binding indicator, ethidium homodimer, and less than 5% of cells were labeled in all regions of the villi and crypt epithelia at 24 h ex vivo. This tissue slice model demonstrated intact myenteric and submucosal neuronal plexuses and functional interstitial cells of Cajal to the extent that nonstimulated, segmental contractions occurred for up to 48 h ex vivo. To detect changes in physiological responses, slices were also assessed for segmental contractions in the presence and absence of antibiotic treatment, which resulted in slices with lesser or greater amounts of commensal bacteria, respectively. Segmental contractions were significantly greater in slices without antibiotics and increased native microbiota. This model renders mechanisms of neuroimmune-microbiome interactions in a complex gut environment available to direct observation and controlled perturbation.
Enteral but not parenteral antibiotics enhance gut function and prevent necrotizing enterocolitis in formula-fed newborn preterm pigs
Authors: Birck Malene M. , Nguyen Duc Ninh , Cilieborg Malene S. , Kamal Shamrulazhar S. , Nielsen Dennis S. , Damborg Peter , Olsen John E. , Lauridsen Charlotte , Sangild Per T. , Thymann Thomas
Published: December 2015

Preterm infants are susceptible to infection and necrotizing enterocolitis (NEC) and are often treated with antibiotics. Simultaneous administration of enteral and parenteral antibiotics during the first days after preterm birth prevents formula-induced NEC lesions in pigs, but it is unknown which administration route is most effective. We hypothesized that only enteral antibiotics suppress gut bacterial colonization and NEC progression in formula-fed preterm pigs. Caesarean-delivered preterm pigs (90–92% of gestation) were fed increasing amounts of infant formula from birth to day 5 and given saline (CON) or antibiotics (ampicillin, gentamicin, and metronidazole) via the enteral (ENT) or parenteral (PAR) route ( n = 16–17). NEC lesions, intestinal morphology, function, microbiology, and inflammatory mediators were evaluated. NEC lesions were completely prevented in ENT pigs, whereas there were high incidences of mild NEC lesions (59–63%) in CON and PAR pigs ( P < 0.001). ENT pigs had elevated intestinal weight, villus height/crypt depth ratio, and goblet cell density and reduced gut permeability, mucosal adherence of bacteria, IL-8 levels, colonic lactic acid levels, and density of Gram-positive bacteria, relative to CON pigs ( P < 0.05). Values in PAR pigs were intermediate with few affected parameters (reduced lactic acid levels and density and adherence of Gram-positive bacteria, relative to CON pigs, P < 0.05). There was no evidence of increased antimicrobial resistance following the treatments. We conclude that enteral, but not parenteral, administration of antibiotics reduces gut bacterial colonization, inflammation, and NEC lesions in newborn, formula-fed preterm pigs. Delayed colonization may support intestinal structure, function, and immunity in the immediate postnatal period of formula-fed preterm neonates.
Abstract A13: Endothelial podosome rosettes regulate vascular branching in tumor angiogenesis
Authors: Seano Giorgio , Chiaverina Giulia , Gagliardi Paolo Armando , Blasio Laura di , Puliafito Alberto , Bouvard Claire , Sessa Roberto , Tarone Guido , Sorokin Lydia , Helley Dominique , Jain Rakesh K. , Serini Guido , Bussolino Federico , Primo Luca
Published: December 2015

The mechanism, by which angiogenic endothelial cells break the physical barrier of vascular basement membrane and consequently sprout, forming new vessels in mature tissues, is unclear. We discovered that angiogenic endothelium is characterized by the presence of functional podosome rosettes. These extracellular matrix-degrading and adhesive structures are precursor of de novo branching points and represent a key event in the formation of new blood vessels. VEGF-A stimulation induces the formation of endothelial podosome rosettes by up-regulating integrin α6β1; in contrast, the binding of α6β1 integrin to vascular basement membrane laminin impairs the formation of podosome rosettes by restricting α6β1 integrin to focal adhesions and hampering its translocation to podosomes. Inhibition of podosome rosette formation by α6β1 blockade or genetic deletion affects vessel branching. The control exerted by α6β1 integrin on tumor blood vessels branching as a consequence of podosome rosette inhibition in endothelial cells recalls the previously proposed notion of vascular normalization by suggesting endothelial podosome rosettes as a new target to normalize tumor vasculature.

Citation Format: Giorgio Seano, Giulia Chiaverina, Paolo Armando Gagliardi, Laura di Blasio, Alberto Puliafito, Claire Bouvard, Roberto Sessa, Guido Tarone, Lydia Sorokin, Dominique Helley, Rakesh K. Jain, Guido Serini, Federico Bussolino, Luca Primo. Endothelial podosome rosettes regulate vascular branching in tumor angiogenesis. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Angiogenesis and Vascular Normalization: Bench to Bedside to Biomarkers; Mar 5-8, 2015; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl):Abstract nr A13.
Abstract A25: Molecular characterization of cyclin-dependent kinase 1 pathway in newly established epithelial ovarian cancer cell lines
Authors: Cho Hanbyoul , Sabrgaliyeva Assel , Yang Woo Kyeom , Kim Sol , Shin Ha-Yeon , Lee Eun Ju , Kim Jae-hoon
Published: December 2015

Background: Epithelial ovarian cancer (EOC) is a heterogeneous gynecologic tumor. To better understand the biological pathways involved in the development of EOC, we performed mRNA expression profiling on newly established EOC cell lines and studied the common characteristics in EOC cells.

Methods: Five newly established EOC cell lines were characterized by transcript profiling. Microarray analyses were validated using quantitative real-time PCR (RT-PCT), immunoblotting, and GEO dataset. Subsequently, we investigated the possible functions of S100A14 in EOC by small interfering RNA (siRNA) approaches.

Results: A total of 239 genes represented by 24,957 probes were identified from microarray analysis, of which 98 and 141 genes are up- (> 2 fold, p < 0.05) or down-regulated (< 3 fold, p < 0.05), respectively, in EOC cell lines compare to human ovarian surface epithelial cells. Interestingly, our data showed upregulation of 11 genes which are associated with the CDK1 pathway. In RT-PCR and immunoblotting, Cdk1 and cyclin B1 expression was significantly increased in EOC cells and tissues compared to benign or normal ovarian surface epitheliums.

Cdk1 knockdown by siCdk1 or Cdk1 inhibitor, RO-3306, inhibited cell growth and arrested the cell cycle at G2/M phase with increased pro-apoptotic signals and decreased anti-apoptotic signals.

Conclusions: These results suggest that elevated expression of Cdk1/cyclinB1 is important to EOC development and progression, providing new insight into the biology of EOC.

Citation Format: Hanbyoul Cho, Assel Sabrgaliyeva, Woo Kyeom Yang, Sol Kim, Ha-Yeon Shin, Eun Ju Lee, Jae-hoon Kim. Molecular characterization of cyclin-dependent kinase 1 pathway in newly established epithelial ovarian cancer cell lines. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Angiogenesis and Vascular Normalization: Bench to Bedside to Biomarkers; Mar 5-8, 2015; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl):Abstract nr A25.
Abstract B05: An immune modulatory role for the Fes protein tyrosine kinase
Authors: Alotaibi Faizah Mesfer , Zhang Connie , Basta Sam , Greer Peter A.
Published: December 2015

The Fes protein tyrosine kinase is abundantly expressed in phagocytic immune cells, including tumor associated macrophages. In addition to the C-terminal kinase and central SH2 domains, Fes contains an N-terminal F-BAR domain that confers unique membrane binding and bending properties which implicate it in the regulation of dynamic membrane-cytoskeletal remodeling processes associated with phagocytosis, receptor endocytosis, vesicular trafficking and cross-talk between receptor systems. In addition to roles in signaling downstream from a variety of cytokine receptors, including those for IL-3, -4, -5, and GM-CSF, Fes has also been implicated in regulating signaling downstream from the TLR-4 receptor for LPS. Fes knockout mice display enhanced sensitivity to LPS, and this was shown to be associated with increased NF-κB signaling and TNFα production from Fes-/- macrophages. Recent studies have also implicated Fes in cross-talk between MHC-I and the NF-κB and IRF-3 pathways in macrophages. Activation of these pro-inflammatory NF-κB and IRF-3 pathways so called Signal 3 - in antigen presenting cells (APCs) is associated with more robust activation of T cells. Interestingly, tumor onset in the MMTV-Neu transgenic mouse model of breast cancer is significantly delayed in Fes-/-, and this was associated with increased frequency of T and B lymphocytes in the premalignant mammary glands.

These observations suggest that Fes contributes to the regulation of signaling downstream from the MHC-I and TLR receptors such that Fes disruption in TLR-engaged APCs would lead to enhanced activation of CD8+ T cells through increased Signal 3 from the T-cell receptor-engaged APCs.

Using a lymphocytic choriomeningitis virus (LCMV) model of immune activation we show that LPS stimulated Fes-/- macrophages promote more robust activation of CD8+ T cells than Fes+/+ macrophages. Furthermore, LPS stimulated Fes-/- macrophages show increased phosphorylation of TBK-1 and IRF-3. We also observed that Fes co-localizes with MHC-I in dynamic vesicular structures within macrophages. These observations are consistent with a model where Fes regulates Signal 3 in APCs through roles in cross-talk between MHC-I and the NF-kB and IRF-3 signaling pathways. This suggests that Fes plays an immune checkpoint role at the level of Signal 3, and that Fes inhibition could promote tumor immunity through increased Signal 3 driven T cell activation.

Citation Format: Faizah Mesfer Alotaibi, Connie Zhang, Sam Basta, Peter A. Greer. An immune modulatory role for the Fes protein tyrosine kinase. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Angiogenesis and Vascular Normalization: Bench to Bedside to Biomarkers; Mar 5-8, 2015; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl):Abstract nr B05.
Abstract B15: In vivo analysis of repeated siRNA silencing on protein expression levels
Authors: Aliabadi Hamidreza Montazeri , Mahdipoor Parvin , Uludag Hasan
Published: December 2015

An ever increasing number of anticancer drugs are under investigation, and enter the clinical setting targeting different tumor survival and/or growth mechanisms; however, development of resistance against the mechanism of action of even the most promising drugs seems to be inevitable and remains a major concern. RNA interference (RNAi) have shown considerable promise in silencing proteins involved in tumor vascularization, as well as intracellular pathways involved in cell proliferation and/or survival. Despite the apparent efficacy of this powerful tool (especially in vitro), our information about the factors affecting siRNA efficiency and the possibility of resistance development against the silencing effect are limited at best. This study focused on determining the possibility of resistance development against siRNA treatment as a result of repeated exposure to siRNA in vivo. Our preliminary experiments in vitro revealed an unaffected siRNA cellular internalization and reproducible silencing efficiency of selected targets. The expression level of other mediators involved in breast cancer cell survival and proliferation (notably survivin, JUN, JAK2, NFkB and STAT3) were, however, altered in siRNA treated cells. In vivo experiments in a xenograft model demonstrated a similar silencing efficiency at the mRNA level after each repeated dose, with little, if any, apparent resistance development to the siRNA therapy. We conclude that cells would respond to repeated siRNA treatments in a similar fashion after a temporary initial alteration.

Citation Format: Hamidreza Montazeri Aliabadi, Parvin Mahdipoor, Hasan Uludag. In vivo analysis of repeated siRNA silencing on protein expression levels. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Angiogenesis and Vascular Normalization: Bench to Bedside to Biomarkers; Mar 5-8, 2015; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl):Abstract nr B15.
Abstract PR04: Intravital imaging of cytolytic activity reveals how poor tumor vascularization and hypoxia limit tumor rejection
Authors: Shakhar Guy , Manaster Yoav , Feferman Tali
Published: December 2015

Background: Immunotherapies that harness cytotoxic T lymphocytes (CTLs) to eradicate solid tumors, melanoma in particular, have re-emerged as potent clinical options in recent years. Improving the dispersion and cytotoxicity of intratumoral CTLs may further benefit patients. One of the obstacles to successful treatment is thought to be poor vascularization of tumors, which leads to sub-optimal tumor infiltration. In contrast, the effects of disrupted vascularization and the hypoxia it introduces on CTL migration and function within tumors have not been investigated.

Methods: To study whether tumor vascularity can shape anti-tumor CTL responses, we combined immunohistochemistry of human melanoma samples with dynamic two-photon imaging within B16 melanoma tumors expressing ovalbumin (OVA) in live mice.

Results: In patients, CD8 T cells were twofold denser around peripheral vessels than around deep vessels, and fourfold denser than in avascular areas. This pattern was clearest in poorly-infiltrated tumors. Correspondingly, in mice, adoptively transferred OT-I CTLs, specific for OVA, preferentially infiltrated B16-OVA tumors through peripheral ICAM1+ vessels. CTLs remained initially at their entry areas, crawling vigorously only within 50µm of flowing blood vessels. These dense CTLs swarms efficiently cleared perivascular areas of tumor cells but left avascular areas intact. In contrast, Foxp3+ regulatory T cells could venture farther away from blood vessels. Target killing was antigen-specific, contact-dependent and perforin-mediated, and required multiple CTLs to engage one target cell to be effective. Several findings converge to suggest that perivascular CTL confinement is oxygen-dependent: occlusion of intratumoral blood vessels resulted in instantaneous and reversible arrest of CTLs. Immunohistology indicated that CTLs avoided hypoxic tumor areas. Hypoxia and mitochondrial uncoupling interfered with CTL migration in vitro and, finally, highly vascular tumors grown in a matrix containing the pro-angiogenic factor bFGF were more easily rejected after CTL injection, displayed delayed growth in untreated mice but were not affected in immunodeficient mice. Although such normalized tumors attracted normal numbers of CTLs, these cells migrated more freely within the tumors.

Conclusions: We propose that CTLs depend on oxygen for intratumoral locomotion and cytolysis. Incomplete vascularization promotes patchy infiltration and tumor escape; tumor immunotherapy may thus benefit from an extensive and functional vasculature.

Citation Format: Guy Shakhar, Yoav Manaster, Tali Feferman. Intravital imaging of cytolytic activity reveals how poor tumor vascularization and hypoxia limit tumor rejection. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Angiogenesis and Vascular Normalization: Bench to Bedside to Biomarkers; Mar 5-8, 2015; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl):Abstract nr PR04.
Can Nonnative Speakers Reduce English Vowels in a Native-Like Fashion? Evidence from L1-Spanish L2-English Bilinguals
Authors: Rallo Fabra Lucrecia
Published: December 2015

This paper investigates the production of English unstressed vowels by two groups of early (ESp) and late Spanish (LSp) bilinguals and a control group of native English (NE) monolinguals. Three acoustic measurements were obtained: duration and intensity ratios of unstressed to stressed vowels, normalized vowel formants and euclidean distances. Both groups of bilinguals showed significantly fewer differences in duration between stressed and unstressed vowels than the NE monolinguals. Intensity differences depended on whether the stress pattern of the target English words matched the stress pattern of their Spanish cognates. As for vowel quality, the early bilinguals reduced the unstressed vowels, which clustered around the midcenter area of the vowel space, in the same fashion as the NE monolinguals, suggesting that vowel reduction might be operating at the phonological level. However, the late bilinguals showed a context-dependent, phonetic-level pattern with vowels that were more peripheral in the vowel space.
The Diagnostic Value of Neuropathy Symptom and Change Score, Neuropathy Impairment Score and Michigan Neuropathy Screening Instrument for Diabetic Peripheral Neuropathy
Authors: Xiong Qian , Lu Bing , Ye Hongying , Wu Xia , Zhang Tiansong , Li Yiming
Published: December 2015

Aims: This study aims to evaluate the diagnostic capabilities of neuropathy symptom and change (NSC) score, neuropathy impairment score (NIS) and Michigan neuropathy screening instrument (MNSI) in diagnosing diabetic peripheral neuropathy (DPN). Methods: A total of 131 patients with type II diabetes received NSC, NIS and MNSI scoring systems. Electromyography/nerve conduction velocity (EMG/NCV) test was taken as gold standard. Correlations between EMG/NCV test and the 3 scorings, and their sensitivity, specificity, positive and negative predictive values, accuracy and kappa (κ) value were analyzed. Results: The prevalence of DPN was 43.5% according to EMG/NCV findings. EMG/NCV test was significantly positive correlated with all the 3 scorings, highest with NIS scoring (r = 0.653, p < 0.001). Compared with EMG/NCV test, NSC score was most sensitive (85.96%) but least specific (77.03%); NIS score had lower sensitivity (59.65%) but best specificity (98.65%) and accuracy (81.68%). Both had high concordance with EMG/NCV test (κ = 0.61). Sensitivity, specificity and accuracy of MNSI were highest (70.18, 98.65 and 80.15%) at the cutoff values of >1.0, >2.5 and >1.5, respectively (κ = 0.58). Conclusions: Both NSC and NIS were accurate and reliable diagnostic methods for DPN. The combined application of NSC and NIS was recommended in DPN diagnosis.