bluuebunny/crossref_metadata_2025 huggingface dataset:
Prolonged adenosine triphosphate infusion and exercise hyperemia in humans
Authors: Shepherd John R. A. , Joyner Michael J. , Dinenno Frank A. , Curry Timothy B. , Ranadive Sushant M.
Published: July 2016

In humans, intra-arterial ATP infusion in limbs mimics many features of exercise hyperemia. However, it remains unknown whether ATP can evoke the prolonged vasodilation seen during exercise. Therefore, we addressed two questions during a continuous 3-h brachial artery infusion of ATP [20 μg·100 ml forearm volume (FAV)−1·min−1]: 1) would skeletal muscle blood flow remain robust or wane over time (tachyphylaxis); and 2) would the hyperemic response to moderate-intensity exercise performed during the ATP administration be blunted compared with that during control (saline) infusion. Nine participants (25 ± 1 yr) performed one trial consisting of seven bouts of rhythmic handgrip exercise (20 contractions/min at 20% of maximum), two bouts during saline (control), and five bouts during 180 min of continuous ATP infusion. Five minutes of ATP infusion resulted in a 710% increase in forearm vascular conductance (FVC) from control (4.8 ± 0.77 vs. 35.0 ± 5.7 ml·min−1·100 mmHg−1·dl FAV−1, P < 0.05). Contrary to our expectations, FVC did not wane over time with values of 35.0 ± 5.7 and 36.0 ± 7.7 ml·min−1·100 mmHg−1·dl FAV−1 ( P > 0.05), seen prior to the exercise bouts at 5 vs. 150 min, respectively. During superimposed exercise, FVC increased from 35.0 ± 5.7 to 49.6 ± 5.4 ml·min−1·100 mmHg−1·dl FAV−1 at 5 min and 36.0 ± 7.7 to 54.5 ± 5.0 at 150 min ( P < 0.05). Our findings demonstrate ATP vasodilation is prolonged over time without tachyphylaxis; however, exercise hyperemia responses remain intact. Our results challenge the metabolic theory of exercise hyperemia, suggesting a disconnect between matching of blood flow and metabolic demand.
Analysis of haptic information in the cerebral cortex
Authors: Sathian K.
Published: July 2016

Haptic sensing of objects acquires information about a number of properties. This review summarizes current understanding about how these properties are processed in the cerebral cortex of macaques and humans. Nonnoxious somatosensory inputs, after initial processing in primary somatosensory cortex, are partially segregated into different pathways. A ventrally directed pathway carries information about surface texture into parietal opercular cortex and thence to medial occipital cortex. A dorsally directed pathway transmits information regarding the location of features on objects to the intraparietal sulcus and frontal eye fields. Shape processing occurs mainly in the intraparietal sulcus and lateral occipital complex, while orientation processing is distributed across primary somatosensory cortex, the parietal operculum, the anterior intraparietal sulcus, and a parieto-occipital region. For each of these properties, the respective areas outside primary somatosensory cortex also process corresponding visual information and are thus multisensory. Consistent with the distributed neural processing of haptic object properties, tactile spatial acuity depends on interaction between bottom-up tactile inputs and top-down attentional signals in a distributed neural network. Future work should clarify the roles of the various brain regions and how they interact at the network level.
aCGH Analysis to Estimate Genetic Variations among Domesticated Chickens
Authors: Komiyama Tomoyoshi , Lin Mengjie , Ogura Atsushi
Published: July 2016

Chickens have been familiar to humans since ancient times and have been used not only for culinary purposes but also for cultural purposes including ritual ceremonies and traditional entertainment. The various chicken breeds developed for these purposes often display distinct morphological and/or behavioural traits. For example, the JapaneseShamois larger and more aggressive than other domesticated chickens, reflecting its role as a fighting cock breed, whereas JapaneseNaganakidoribreeds, which have long-crowing behaviour, were bred instead for their entertaining and aesthetic qualities. However, the genetic backgrounds of these distinct morphological and behavioural traits remain unclear. Therefore, the question arises as to which genomic regions in these chickens were acted upon by selective pressures through breeding. We compared the entire genomes of six chicken breeds domesticated for various cultural purposes by utilizing array comparative genomic hybridization. From these analyses, we identified 782 regions that underwent insertions, deletions, or mutations, representing man-made selection pressure in these chickens. Furthermore, we found that a number of genes diversified in domesticated chickens bred for cultural or entertainment purposes were different from those diversified in chickens bred for food, such as broilers and layers.
Sparsity-Homotopy Perturbation Inversion Method with Wavelets and Applications to Black-Scholes Model and Todaro Model
Authors: Dou Yixin , Wang Zhihao
Published: July 2016

Sparsity regularization method plays an important role in reconstructing parameters. Compared with traditional regularization methods, sparsity regularization method has the ability to obtain better performance for reconstructing sparse parameters. However, sparsity regularization method does not have the ability to reconstruct smooth parameters. For overcoming this difficulty, we combine a sparsity regularization method with a wavelet method in order to transform smooth parameters into sparse parameters. We use a sparsity-homotopy perturbation inversion method to improve the accuracy and stability and apply the proposed method to reconstruct parameters for a Black-Scholes option pricing model and a Todaro model. Numerical experiments show that the proposed method is convergent and stable.
Prediction of Collapse Scope of Deep-Buried Tunnels Using Pressure Arch Theory
Authors: Wang Yingchao , Jing Hongwen , Zhang Qiang , Luo Ning , Yin Xin
Published: July 2016

Tunnel collapse remains a serious problem in practice. Effective prediction methods on tunnel collapse are necessary for tunnel engineering. In this study, systematic study on the pressure arch was presented to predict tunnel collapse. Multiple factors under different conditions were considered. First, the pressure arch was described as a certain scope in comparison with the lowest pressure arch line. Then, a deep-buried circular tunnel was selected as the investigated object. Its collapse scope was analyzed using the lowest pressure arch line. Meanwhile, the main influence from the ground stress field was considered. Different modes of ground stress fields were investigated in detail. The results indicate that the collapse scope varies with different ground stress fields. Determination on the collapse scope is strongly affected by the judgment standard of the pressure arch. Furthermore, a selected case was analyzed with the pressure arch. The area and the height of tunnel collapse were calculated with multiple factors, including ground stress field, judgment standard, and lateral pressure coefficient. Finally, selected results were compared with relevant previous researches, and reasonable results were obtained. The present results are helpful for further understanding of the tunnel collapse and could provide suitable guidance for tunnel projects.
A Multiconstrained Ascent Guidance Method for Solid Rocket-Powered Launch Vehicles
Authors: Chen Si-Yuan , Xia Qun-Li
Published: July 2016

This study proposes a multiconstrained ascent guidance method for a solid rocket-powered launch vehicle, which uses a hypersonic glide vehicle (HGV) as payload and shuts off by fuel exhaustion. First, pseudospectral method is used to analyze the two-stage launch vehicle ascent trajectory with different rocket ignition modes. Then, constraints, such as terminal height, velocity, flight path angle, and angle of attack, are converted into the constraints within height-time profile according to the second-stage rocket flight characteristics. The closed-loop guidance method is inferred by different spline curves given the different terminal constraints. Afterwards, a thrust bias energy management strategy is proposed to waste the excess energy of the solid rocket. Finally, the proposed method is verified through nominal and dispersion simulations. The simulation results show excellent applicability and robustness of this method, which can provide a valuable reference for the ascent guidance of solid rocket-powered launch vehicles.
Validity of a Smartphone-Based Application for Determining Sprinting Performance
Authors: Stanton Robert , Hayman Melanie , Humphris Nyree , Borgelt Hanna , Fox Jordan , Del Vecchio Luke , Humphries Brendan
Published: July 2016

Recent innovations in smartphone technology have led to the development of a number of applications for the valid and reliable measurement of physical performance. Smartphone applications offer a number of advantages over laboratory based testing including cost, portability, and absence of postprocessing. However, smartphone applications for the measurement of running speed have not yet been validated. In the present study, the iOS smartphone application,SpeedClock, was compared to conventional timing lights during flying 10 m sprints in recreationally active women. Independent samplest-test showed no statistically significant difference betweenSpeedClockand timing lights (t(190)=1.83,p=0.07), while intraclass correlations showed excellent agreement betweenSpeedClockand timing lights (ICC (2,1) = 0.93,p=0.00, 95% CI 0.64–0.97). Bland-Altman plots showed a small systematic bias (mean difference = 0.13 seconds) withSpeedClockgiving slightly lower values compared to the timing lights. Our findings suggestSpeedClockfor iOS devices is a low-cost, valid tool for the assessment of mean flying 10 m sprint velocity in recreationally active females. Systematic bias should be considered when interpreting the results fromSpeedClock.
Comparing the Performance of Popular MEG/EEG Artifact Correction Methods in an Evoked-Response Study
Authors: Haumann Niels Trusbak , Parkkonen Lauri , Kliuchko Marina , Vuust Peter , Brattico Elvira
Published: July 2016

We here compared results achieved by applying popular methods for reducing artifacts in magnetoencephalography (MEG) and electroencephalography (EEG) recordings of the auditory evoked Mismatch Negativity (MMN) responses in healthy adult subjects. We compared the Signal Space Separation (SSS) and temporal SSS (tSSS) methods for reducing noise from external and nearby sources. Our results showed that tSSS reduces the interference level more reliably than plain SSS, particularly for MEG gradiometers, also for healthy subjects not wearing strongly interfering magnetic material. Therefore, tSSS is recommended over SSS. Furthermore, we found that better artifact correction is achieved by applying Independent Component Analysis (ICA) in comparison to Signal Space Projection (SSP). Although SSP reduces the baseline noise level more than ICA, SSP also significantly reduces the signal—slightly more than it reduces the artifacts interfering with the signal. However, ICA also adds noise, or correction errors, to the waveform when the signal-to-noise ratio (SNR) in the original data is relatively low—in particular to EEG and to MEG magnetometer data. In conclusion, ICA is recommended over SSP, but one should be careful when applying ICA to reduce artifacts on neurophysiological data with relatively low SNR.
Coverage of Axillary Lymph Nodes with Tangential Breast Irradiation in Korea: A Multi-Institutional Comparison Study
Authors: Jung Jinhong , Kong Moonkyoo , Kim Su Ssan , Yoon Won Sup
Published: July 2016

Introduction. To evaluate the dose distribution and coverage of axilla using only tangential field for whole breast radiotherapy (RT) at three institutions in Korea.Methods. We used computed tomography (CT) images of nine consecutive 1-2 sentinel lymph node-positive patients who underwent breast conserving surgery and whole breast RT without axillary lymph node (ALN) dissection for clinical T1-2N0 breast cancer. The CT data were transferred to three radiation oncologists in 3 institutions and each radiation oncologist created treatment plans for all nine patients; a total of 27 treatment plans were analyzed.Results. The mean doses delivered to levels I and II were 31.9 Gy (9.9–47.9 Gy) and 22.3 Gy (3.4–47.7 Gy). Ninety-five percent of levels I and II received a mean dose of 11.8 Gy (0.4–43.0 Gy) and 3.0 Gy (0.3–40.0 Gy). The percent volumes of levels I and II covered by 95% of the prescribed dose were only 29.0% (0.2–74.1%) and 11.5% (0.0–70.1%). The dose distribution and coverage of axilla were significantly different between three institutions (p=0.001).Conclusion. There were discrepancies in ALN coverage between three institutions. A standardization of whole breast RT technique through further research with a nationwide scale is needed.
Advanced Genetic Testing Comes to the Pain Clinic to Make a Diagnosis of Paroxysmal Extreme Pain Disorder
Authors: Cannon Ashley , Kurklinsky Svetlana , Guthrie Kimberly J. , Riegert-Johnson Douglas L.
Published: July 2016

Objective. To describe the use of an advanced genetic testing technique, whole exome sequencing, to diagnose a patient and their family with aSCN9Achannelopathy.Setting. Academic tertiary care center.Design. Case report.Case Report. A 61-year-old female with a history of acute facial pain, chronic pain, fibromyalgia, and constipation was found to have a gain of functionSCN9Amutation by whole exome sequencing. This mutation resulted in anSCN9Achannelopathy that is most consistent with a diagnosis of paroxysmal extreme pain disorder. In addition to the patient being diagnosed, four siblings have a clinical diagnosis ofSCN9Achannelopathy as they have consistent symptoms and a sister with a known mutation. For treatment, gabapentin was ineffective and carbamazepine was not tolerated. Nontraditional therapies improved symptoms and constipation resolved with pelvic floor retraining with biofeedback.Conclusion. Patients with a personal and family history of chronic pain may benefit from a referral to Medical Genetics. Pelvic floor retraining with biofeedback should be considered for patients with aSCN9Achannelopathy and constipation.
Functional Design in Rehabilitation: Modular Mechanisms for Ankle Complex
Authors: Aggogeri Francesco , Pellegrini Nicola , Adamini Riccardo
Published: July 2016

This paper is aimed at presenting an innovative ankle rehabilitation device based on a parallel mechanism. A functional analysis and design are described to obtain a device able to guarantee ankle movement while patient’s body remains stationary. Human ankle is a challenging context where a series of joints are highly integrated. The proposed rehabilitation device permits a patient with walking defects to improve his or her gait. The research focuses on plantar-flexion-dorsiflexion movement. The robust design starts from an accurate modelling of ankle movements during walking, assessing motion data from healthy individuals and patients. The kinematics analysis and functional evaluations lead the study and development of the articulated system. In particular, results of simulations support the effectiveness of the current design. A 3D prototype is presented highlighting that the ankle motion is successfully demonstrated.
Plasticity, Swell-Shrink, and Microstructure of Phosphogypsum Admixed Lime Stabilized Expansive Soil
Authors: James Jijo , Pandian P. Kasinatha
Published: July 2016

The study involved utilization of an industrial waste, Phosphogypsum (PG), as an additive to lime stabilization of an expansive soil. Three lime dosages, namely, initial consumption of lime (ICL), optimum lime content (OLC), and less than ICL (LICL), were identified for the soil under study for stabilizing the soil. Along with lime, varying doses of PG were added to the soil for stabilization. The effect of stabilization was studied by performing index tests, namely, liquid limit, plastic limit, shrinkage limit, and free swell test, on pulverized remains of failed unconfined compression test specimens. The samples were also subjected to a microstructural study by means of scanning electron microscope. Addition of PG to lime resulted in improvement in the plasticity and swell-shrink characteristics. The microstructural study revealed the formation of a dense compact mass of stabilized soil.
The Cancer Epidemiology Descriptive Cohort Database: A Tool to Support Population-Based Interdisciplinary Research
Authors: Kennedy Amy E. , Khoury Muin J. , Ioannidis John P.A. , Brotzman Michelle , Miller Amy , Lane Crystal , Lai Gabriel Y. , Rogers Scott D. , Harvey Chinonye , Elena Joanne W. , Seminara Daniela
Published: July 2016

Background: We report on the establishment of a web-based Cancer Epidemiology Descriptive Cohort Database (CEDCD). The CEDCD's goals are to enhance awareness of resources, facilitate interdisciplinary research collaborations, and support existing cohorts for the study of cancer-related outcomes.

Methods: Comprehensive descriptive data were collected from large cohorts established to study cancer as primary outcome using a newly developed questionnaire. These included an inventory of baseline and follow-up data, biospecimens, genomics, policies, and protocols. Additional descriptive data extracted from publicly available sources were also collected. This information was entered in a searchable and publicly accessible database. We summarized the descriptive data across cohorts and reported the characteristics of this resource.

Results: As of December 2015, the CEDCD includes data from 46 cohorts representing more than 6.5 million individuals (29% ethnic/racial minorities). Overall, 78% of the cohorts have collected blood at least once, 57% at multiple time points, and 46% collected tissue samples. Genotyping has been performed by 67% of the cohorts, while 46% have performed whole-genome or exome sequencing in subsets of enrolled individuals. Information on medical conditions other than cancer has been collected in more than 50% of the cohorts. More than 600,000 incident cancer cases and more than 40,000 prevalent cases are reported, with 24 cancer sites represented.

Conclusions: The CEDCD assembles detailed descriptive information on a large number of cancer cohorts in a searchable database.

Impact: Information from the CEDCD may assist the interdisciplinary research community by facilitating identification of well-established population resources and large-scale collaborative and integrative research. Cancer Epidemiol Biomarkers Prev; 25(10); 1392–401. ©2016 AACR.
The Novel Pan-PIM Kinase Inhibitor, PIM447, Displays Dual Antimyeloma and Bone-Protective Effects, and Potently Synergizes with Current Standards of Care
Authors: Paíno Teresa , Garcia-Gomez Antonio , González-Méndez Lorena , San-Segundo Laura , Hernández-García Susana , López-Iglesias Ana-Alicia , Algarín Esperanza M. , Martín-Sánchez Montserrat , Corbacho David , Ortiz-de-Solorzano Carlos , Corchete Luis A. , Gutiérrez Norma C. , Maetos María-Victoria , Garayoa Mercedes , Ocio Enrique M.
Published: July 2016

Purpose: PIM kinases are a family of serine/threonine kinases recently proposed as therapeutic targets in oncology. In the present work, we have investigated the effects of the novel pan-PIM kinase inhibitor, PIM447, on myeloma cells and myeloma-associated bone disease using different preclinical models.

Experimental Design: In vitro/ex vivo cytotoxicity of PIM447 was evaluated on myeloma cell lines and patient samples. Synergistic combinations with standard treatments were analyzed with Calcusyn Software. PIM447 effects on bone cells were assessed on osteogenic and osteoclastogenic cultures. The mechanisms of PIM447 were explored by immunoblotting, qPCR, and immunofluorescence. A murine model of disseminated multiple myeloma was employed for in vivo studies.

Results: PIM447 is cytotoxic for myeloma cells due to cell-cycle disruption and induction of apoptosis mediated by a decrease in phospho-Bad (Ser112) and c-Myc levels and the inhibition of mTORC1 pathway. Importantly, PIM447 demonstrates a very strong synergy with different standard treatments such as bortezomib + dexamethasone (combination index, CI = 0.002), lenalidomide + dexamethasone (CI = 0.065), and pomalidomide + dexamethasone (CI = 0.077). PIM447 also inhibits in vitro osteoclast formation and resorption, downregulates key molecules involved in these processes, and partially disrupts the F-actin ring, while increasing osteoblast activity and mineralization. Finally, PIM447 significantly reduced the tumor burden and prevented tumor-associated bone loss in a disseminated murine model of human myeloma.

Conclusions: Our results demonstrate dual antitumoral and bone-protective effects of PIM447. This fact, together with the very strong synergy exhibited with standard-of-care treatments, supports the future clinical development of this drug in multiple myeloma. Clin Cancer Res; 23(1); 225–38. ©2016 AACR.
Genomic Copy Number Profiling Using Circulating Free Tumor DNA Highlights Heterogeneity in Neuroblastoma
Authors: Chicard Mathieu , Boyault Sandrine , Colmet Daage Leo , Richer Wilfrid , Gentien David , Pierron Gaelle , Lapouble Eve , Bellini Angela , Clement Nathalie , Iacono Isabelle , Bréjon Stéphanie , Carrere Marjorie , Reyes Cécile , Hocking Toby , Bernard Virginie , Peuchmaur Michel , Corradini Nadège , Faure-Conter Cécile , Coze Carole , Plantaz Dominique , Defachelles Anne Sophie , Thebaud Estelle , Gambart Marion , Millot Frédéric , Valteau-Couanet Dominique , Michon Jean , Puisieux Alain , Delattre Olivier , Combaret Valérie , Schleiermacher Gudrun
Published: July 2016

Purpose: The tumor genomic copy number profile is of prognostic significance in neuroblastoma patients. We have studied the genomic copy number profile of cell-free DNA (cfDNA) and compared this with primary tumor arrayCGH (aCGH) at diagnosis.

Experimental Design: In 70 patients, cfDNA genomic copy number profiling was performed using the OncoScan platform. The profiles were classified according to the overall pattern, including numerical chromosome alterations (NCA), segmental chromosome alterations (SCA), and MYCN amplification (MNA).

Results: Interpretable and dynamic cfDNA profiles were obtained in 66 of 70 and 52 of 70 cases, respectively. An overall identical genomic profile between tumor aCGH and cfDNA was observed in 47 cases (3 NCAs, 22 SCAs, 22 MNAs). In one case, cfDNA showed an additional SCA not detected by tumor aCGH. In 4 of 8 cases with a silent tumor aCGH profile, cfDNA analysis revealed a dynamic profile (3 SCAs, 1 NCA). In 14 cases, cfDNA analysis did not reveal any copy number changes. A total of 378 breakpoints common to the primary tumor and cfDNA of any given patient were identified, 27 breakpoints were seen by tumor aCGH, and 54 breakpoints were seen in cfDNA only, including two cases with interstitial IGFR1 gains and two alterations targeting TERT.

Conclusions: These results demonstrate the feasibility of cfDNA copy number profiling in neuroblastoma patients, with a concordance of the overall genomic profile in aCGH and cfDNA dynamic cases of 97% and a sensitivity of 77%, respectively. Furthermore, neuroblastoma heterogeneity is highlighted, suggesting that cfDNA might reflect genetic alterations of more aggressive cell clones. Clin Cancer Res; 22(22); 5564–73. ©2016 AACR.

See related commentary by Janku and Kurzrock, p. 5400
PARP Inhibitor Activity Correlates with SLFN11 Expression and Demonstrates Synergy with Temozolomide in Small Cell Lung Cancer
Authors: Lok Benjamin H. , Gardner Eric E. , Schneeberger Valentina E. , Ni Andy , Desmeules Patrice , Rekhtman Natasha , de Stanchina Elisa , Teicher Beverly A. , Riaz Nadeem , Powell Simon N. , Poirier John T. , Rudin Charles M.
Published: July 2016

Purpose: PARP inhibitors (PARPi) are a novel class of small molecule therapeutics for small cell lung cancer (SCLC). Identification of predictors of response would advance our understanding, and guide clinical application, of this therapeutic strategy.

Experimental Design: Efficacy of PARP inhibitors olaparib, rucaparib, and veliparib, as well as etoposide and cisplatin in SCLC cell lines, and gene expression correlates, was analyzed using public datasets. HRD genomic scar scores were calculated from Affymetrix SNP 6.0 arrays. In vitro talazoparib efficacy was measured by cell viability assays. For functional studies, CRISPR/Cas9 and shRNA were used for genomic editing and transcript knockdown, respectively. Protein levels were assessed by immunoblotting and immunohistochemistry (IHC). Quantitative synergy of talazoparib and temozolomide was determined in vitro. In vivo efficacy of talazoparib, temozolomide, and the combination was assessed in patient-derived xenograft (PDX) models.

Results: We identified SLFN11, but not HRD genomic scars, as a consistent correlate of response to all three PARPi assessed, with loss of SLFN11 conferring resistance to PARPi. We confirmed these findings in vivo across multiple PDX and defined IHC staining for SLFN11 as a predictor of talazoparib response. As temozolomide has activity in SCLC, we investigated combination therapy with talazoparib and found marked synergy in vitro and efficacy in vivo, which did not solely depend on SLFN11 or MGMT status.

Conclusions: SLFN11 is a relevant predictive biomarker of sensitivity to PARP inhibitor monotherapy in SCLC and we identify combinatorial therapy with TMZ as a particularly promising therapeutic strategy that warrants further clinical investigation. Clin Cancer Res; 23(2); 523–35. ©2016 AACR.
Enhanced Eryptosis Following Exposure to Dolutegravir
Authors: Al Mamun Bhuyan Abdulla , Signoretto Elena , Bissinger Rosi , Lang Florian
Published: July 2016

Background/Aims: The viral integrase enzyme inhibitor dolutegravir is utilized for the treatment of immunodeficiency virus (HIV) infection. Knowledge on cytotoxicity of dolutegravir is limited. The present study thus explored, whether dolutegravir is able to trigger suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Cellular mechanisms involved in the triggering of eryptosis include increase of cytosolic Ca2+ activity ([Ca2+]i), oxidative stress, ceramide, and activation of protein kinase C, p38 kinase, casein kinase, and caspases. The present study explored, whether Dolutegravir induces eryptosis and, if so, to gain insight into cellular mechanisms involved. Methods: Utilizing flow cytometry, phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ROS formation from DCFDA dependent fluorescence, and ceramide abundance utilizing specific antibodies. Hemolysis was quantified from haemoglobin concentration in the supernatant. Results: A 48 hours exposure of human erythrocytes to dolutegravir significantly increased the percentage of annexin-V-binding cells (≥ 4.8 µM), significantly increased hemolysis (19.1 µM), but did not significantly modify forward scatter. Dolutegravir significantly increased Fluo3-fluorescence (≥ 4.8 µM), DCFDA fluorescence (19.1 µM) and ceramide abundance (19.1 µM). The effect of dolutegravir on annexin-V-binding was significantly blunted by removal of extracellular Ca2+, but was not significantly modified by protein kinase C inhibitor staurosporine (1 µM), p38 kinase inhibitor SB203580 (2 µM), casein kinase inhibitor D4476 (10 µM) or pancaspase inhibitor zVAD (10 µM). Conclusions: Dolutegravir triggers cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect at least in part due to Ca2+ entry, ceramide formation and oxidative stress.
Storage of Erythrocytes Induces Suicidal Erythrocyte Death
Authors: Lang Elisabeth , Pozdeev Vitaly I. , Xu Haifeng C. , Shinde Prashant V. , Behnke Kristina , Hamdam Junnat M. , Lehnert Erik , Scharf Rüdiger E. , Lang Florian , Häussinger Dieter , Lang Karl S. , Lang Philipp A.
Published: July 2016

Background/Aims: Similar to apoptosis of nucleated cells, red blood cells (RBC) can undergo suicidal cell death - called eryptosis. It is characterized by cell shrinkage and phosphatidylserine translocation. Eryptosis is triggered by an increase of intracellular calcium concentration due to activation of nonselective cation channels. The cation channels and consequently eryptosis are inhibited by erythropoietin. Eryptotic RBC are engulfed by macrophages and thus rapidly cleared from circulating blood. In this study, we explored whether storage of RBC influences the rate of eryptosis. Methods: Flow cytometry was employed to quantify phosphatidylserine exposing erythrocytes from annexin V binding and cytosolic Ca2+ activity from Fluo-3 fluorescence. Clearance of stored murine RBC was tested by injection of carboxyfluorescein succinimidyl ester (CFSE)-labelled erythrocytes. Results: Storage for 42 days significantly increased the percentage of phosphatidylserine exposing and haemolytic erythrocytes, an effect blunted by removal of extracellular calcium. Phosphatidylserine exposure could be inhibited by addition of erythropoietin. Upon transfusion, the clearance of murine CFSE-labelled RBC from circulating blood was significantly higher following storage for 10 days when compared to 2 days of storage. Conclusion: Storage of RBC triggers eryptosis by Ca2+ and erythropoietin sensitive mechanisms.
Distinct Gene Expression Patterns Defining Human Osteoblasts' Response to BMP2 Treatment: Is the Therapeutic Success All a Matter of Timing?
Authors: Ehnert Sabrina , Aspera-Werz Romina Haydée , Freude Thomas , Reumann Marie Karolina , Ochs Björn Gunnar , Bahrs Christian , Schröter Steffen , Wintermeyer Elke , Nussler Andreas Klaus , Pscherer Stefan
Published: July 2016

Background: Bone morphogenetic proteins (BMPs) play a key role in bone formation. Local application of BMP2 (Dibotermin alfa) supports bone formation when applied to complex fractures. However, up to 33% of patients do not respond to this therapy. Purpose: Aiming to investigate whether inter-individual responses to BMP2 treatment can be predicted by gene expression patterns, we investigated the effect of BMP2 on primary human osteoblasts and THP-1 cell-derived osteoclasts from 110 donors. Methods: Osteoblasts were obtained by collagenase digestion of spongy bone tissues. Osteoclasts were differentiated from THP-1 cells using the conditioned media of the osteoblasts. Viability was determined by resazurin conversion. As functional characteristics AP and Trap5B activity were measured. Gene expression levels were determined by RT-PCR in 21 of the 110 evaluated donors and visualized by electrophoresis. Results: Based on our data, we could classify three response groups: (i) In 51.8% of all donors, BMP2 treatment induced osteoblast function. These donors strongly expressed the BMP2 inhibitor Noggin (NOG), the alternative BMP2 receptors repulsive guidance molecule B (RGMb) and activin receptor-like kinase 6 (Alk6), as well as the Wnt inhibitor sclerostin (SOST). (ii) In 17.3% of all donors, BMP2 treatment induced viability. In these donors, the initial high SOST expression significantly dropped with BMP2 treatment. (iii) 30.9% of all donors were not directly affected by BMP2 treatment. These donors expressed high levels of the pseudoreceptor BMP and activin membrane-bound inhibitor (BAMBI) and lacked SOST expression. In all donors, SOST expression correlated directly with receptor activator of NF-κB ligand (RANKL) expression, defining the cells' potential to stimulate osteoclastogenesis. Conclusions: Our data identified three donor groups profiting from BMP2 treatment either directly via stimulation of osteoblast function or viability and/or indirectly via inhibition of osteoclastogenesis, depending on their expression of BAMBI, SOST, NOG, and RANKL. On the basis of patients' respective expression profiles, the clinical application of BMP2 as well as its timing might be modified in order to better fit the patients' needs to promote bone formation or to inhibit bone resorption.
Progranulin Levels in Plasma and Cerebrospinal Fluid in Granulin Mutation Carriers
Authors: Meeter Lieke H.H. , Patzke Holger , Loewen Gordon , Dopper Elise G.P. , Pijnenburg Yolande A.L. , van Minkelen Rick , van Swieten John C.
Published: July 2016

Background: Pathogenic mutations in the granulin gene (GRN) are causative in 5-10% of patients with frontotemporal dementia (FTD), mostly leading to reduced progranulin protein (PGRN) levels. Upcoming therapeutic trials focus on enhancing PGRN levels. Methods: Fluctuations in plasma PGRN (n = 41) and its relationship with cerebrospinal fluid (CSF, n = 32) and specific single nucleotide polymorphisms were investigated in pre- and symptomatic GRN mutation carriers and controls. Results: Plasma PGRN levels were lower in carriers than in controls and showed a mean coefficient of variation of 5.3% in carriers over 1 week. Although plasma PGRN correlated with CSF PGRN in carriers (r = 0.54, p = 0.02), plasma only explained 29% of the variability in CSF PGRN. rs5848, rs646776 and rs1990622 genotypes only partly explained the variability of PGRN levels between subjects. Conclusions: Plasma PGRN is relatively stable over 1 week and therefore seems suitable for treatment monitoring of PGRN-enhancing agents. Since plasma PGRN only moderately correlated with CSF PGRN, CSF sampling will additionally be needed in therapeutic trials.
Corneal Ring Infiltrates Caused by Serratia marcescens in a Patient with Human Immunodeficiency Virus
Authors: Chaidaroon Winai , Supalaset Sumet
Published: July 2016

Purpose: To describe corneal ring infiltrates caused by Serratia marcescens in a patient with human immunodeficiency virus (HIV-1) who wore contact lenses. Methods: A case study of a patient with keratitis due to an infection caused by S. marcescens and exhibiting corneal ring infiltrates was reviewed for history, clinical manifestation, microscopic study, and management. Results: A 29-year-old man who had a history of contact lens wear and HIV-1 infection was admitted to hospital because of blurred vision, redness, and corneal infiltrates in the shape of a ring in the left eye. The visual acuity (VA) in both eyes was hand movement (uncorrected). Corneal scrapings were performed. The culture results of the corneal specimens revealed S. marcescens. The culture results of the contact lens disclosed the same organism. The corneal ulcer responded well to treatment with topical gentamycin sulfate 14 mg/ml. The final VA remained hand movement. Conclusions: S. marcescens can cause ring infiltrates in a HIV-1 patient who wears contact lenses. The treatment result for S. marcescens keratitis in a HIV-1 patient who wore contact lenses was favorable after intensive use of fortified topical antibiotics.
Phosphate Binding Therapy to Lower Serum Fibroblast-Growth-Factor-23 Concentrations in Chronic Kidney Disease: Rationale and Study Design of the Sevelamer on FGF23 Trial (SoFT)
Authors: Adema Aaltje Y. , de Jong Maarten A. , de Borst Martin H. , ter Wee Pieter M. , Vervloet Marc G. ,   for the NIGRAM Consortium
Published: July 2016

Background: Increased levels of phosphate and fibroblast growth factor-23 (FGF23) are strong predictors of cardiovascular morbidity and mortality in patients with chronic kidney disease (CKD). Preliminary data suggest that interventions lowering gastro-intestinal phosphate uptake lowers serum FGF23 concentrations and improves cardiovascular risk and subsequently survival. However, data are lacking about the magnitude of effects, the effect in different stages of CKD and whether there is a dose-effect relationship. Methods: Therefore, the Sevelamer on FGF23 Trial (SoFT) is designed as an open-label, single-arm, clinical pilot study aiming to demonstrate the feasibility of a phosphate-restricted diet in combination with the phosphate binder sevelamer to induce an effective, predictable and sustained decrease in FGF23 level in patients with an estimated glomerular filtration rate (eGFR) of 15-90 or >90 ml/min/1.73 m2 with proteinuria >1.0 g in 24 h urine collection, despite optimally dosed RAAS blockade, without inducing hypophosphatemia using a forced uptitration treatment regimen aimed at restricting phosphate uptake.
Preclinical Evaluation of AdVince, an Oncolytic Adenovirus Adapted for Treatment of Liver Metastases from Neuroendocrine Cancer
Authors: Yu Di , Leja-Jarblad Justyna , Loskog Angelica , Hellman Per , Giandomenico Valeria , Oberg Kjell , Essand Magnus
Published: July 2016

Cancer immunotherapy is becoming a cornerstone in the clinical care of cancer patients due to the breakthrough trials with immune checkpoint blockade antibodies and chimeric antigen receptor T cells. The next breakthrough in cancer immunotherapy is likely to be oncolytic viruses engineered to selectively kill tumor cells and deceive the immune system to believe that the tumor is a foreign entity that needs to be eradicated. We have developed AdVince, an oncolytic adenovirus for treatment of liver metastases from neuroendocrine tumor (NET). AdVince includes the gene promoter from human chromogranin A for selective replication in neuroendocrine cells, miR122 target sequences for reduced liver toxicity, and a cell-penetrating peptide in the capsid for increased infectivity of tumor cells and optimized spread within tumors. This paper describes the preclinical evaluation of AdVince on freshly isolated human gastrointestinal NET cells resected from liver metastases and freshly isolated human hepatocytes as well as in fresh human blood. AdVince selectively replicates in and kills NET cells. Approximately 73-fold higher concentration of AdVince is needed to induce a similar level of cytotoxicity in NET cells as in hepatocytes. AdVince did not activate complement or induce considerable amount of proinflammatory cytokines or chemokines in human blood. The data presented herein indicate that AdVince can be safely evaluated in a phase I/IIa clinical trial for patients with liver-dominant NET.
LMWH in the prevention of preeclampsia and fetal growth restriction in women without thrombophilia
Authors: Mastrolia Salvatore , Novack Lena , Thachil Jecko , Rabinovich Anat , Pikovsky Oleg , Klaitman Vered , Loverro Giuseppe , Erez Offer
Published: July 2016

Placental mediated pregnancy complications such as preeclampsia and fetal growth restriction (FGR) are common, serious, and associated with increased morbidity and mortality. We conducted a systematic review and meta-analysis to determine the effect of treatment with low-molecular-weight heparins (LMWHs) for secondary prevention of these complications in non thrombophilic women. We searched the electronic databases PubMed, Scopus, and Cochrane Library for randomised controlled trials addressing this question. Five studies including 403 patients met the inclusion criteria, 68 developed preeclampsia and 118 FGR. The studies were very heterogeneous in terms of inclusion criteria, LMWH preparation, and dosage. Meta-analyses were performed using random-effect models. The overall use of LMWHs was associated with a risk reduction for preeclampsia (Relative risk (RR) 0.366; 95 % confidence interval (CI), 0.219–0.614) and FGR (RR 0.409; 95 % CI, 0.195–0.932) vs. no treatment. From the data available for analysis it appears that the use of Dalteparin is associated with a risk reduction for preeclampsia (p=0.002) and FGR (p<0.001); while Enoxaparin is associated with risk reduction for preeclampsia (p=0.013) but not for FGR (p=0.3). In spite of the small number of studies addressing the research question, and the high variability among them, our meta-analysis found a modest beneficial effect of LMWH for secondary prevention of preeclampsia and FGR. Further studies are needed to address these questions before a definite conclusion can be reached.

Supplementary Material to this article is available online at www.thrombosis-online.com.
Adjusting enoxaparin dosage according to anti-FXa levels and pregnancy outcome in thrombophilic women
Authors: Nachum Zohar , Gavish Israel , Romano Shabtai , Braverman Meirav , Garmi Gali , Salim Raed
Published: July 2016

Women with thrombophilias and previous placenta-mediated pregnancy complications (PMPC) have an increased risk of both recurrent PMPC and venous thromboembolism (VTE) during subsequent pregnancies. We aimed to examine whether enoxaparin dose adjusted according to anti-factor Xa levels compared to a fixed dose would reduce the risk of PMPC in subsequent pregnancies. In a randomised trial, conducted at a single teaching hospital, pregnant women with thrombophilia and previous PMPC were enrolled in a 1:1 ratio to either a fixed dose of 40 mg daily enoxaparin or adjusted dose according to anti-factor Xa plasma levels. The primary outcome was a composite that included any of the following: pregnancy loss after enrollment, pre-eclampsia, birthweight <10th percentile, placental abruption, or VTE. Overall, 144 women were needed to detect a decrease of 20 % in the incidence of the composite outcome among the adjusted dose group. Between 2009 and 2015, 144 women consented; four in the fixed-dose group were excluded. Overall, 66 and 74 in the fixed- and adjusted-dose groups, respectively, were included. Demographic and obstetric characteristics were comparable. Composite outcome occurred in 12 (18.2 %) and 20 (27.0 %) women in the fixed- and adjusted-dose groups, respectively (p=0.24). Gestational age at delivery, preterm births, and birthweights were similar between the two groups. In conclusion, dose of enoxaparin adjusted according to anti-factor Xa levels compared to fixed dose, does not reduce the risk of PMPC recurrence in thrombophilic women.

ClinicalTrial Registration: ClinicalTrials.gov, www.clinicaltrials.gov, NCT01068795.
Patient-independent variables affecting the assessment of aspirin responsiveness by serum thromboxane measurement
Authors: Petrucci Giovanna , Rizzi Alessandro , Cavalca Viviana , Habib Aida , Pitocco Dario , Veglia Fabrizio , Ranalli Paola , Zaccardi Francesco , Pagliaccia Francesca , Tremoli Elena , Patrono Carlo , Rocca Bianca
Published: July 2016

The serum TXB2 (sTXB2) assay reflects the pharmacodynamics of platelet inhibition by low-dose aspirin. However, different studies reported variable sTXB2 values. sTXB2 assay requires whole blood incubation at 37 °C as a condition for optimal thrombin generation, arachidonic acid release and its metabolism by platelet cyclooxygenase-1 to form TXA2. Access to 37 °C incubation may be variably delayed, and different methods to quantitate sTXB2 may contribute to variable results between different Centers. We investigated whether delaying 37 °C incubation and/or analytical issues affect sTXB2 concentrations, biasing the assessment of aspirin responsiveness. Sixty-eight samples from 54 volunteers, on- and off-aspirin, were incubated at 37 °C immediately after sampling (reference sample) or after 5, 10, 15, 20, 30 or 60 minutes at room temperature (RT); 8 samples remained at RT 60 minutes, without subsequent incubation; 314 sera were measured by enzyme immunoassay (EIA) and liquid chromatography-tandem mass-spectrometry (LC/MS-MS) methods. sTXB2 concentrations decreased exponentially as a function of the delay before 37 °C incubation, ranging from 94 ± 11 % at 5 minutes to 23 ± 22 % of the reference sample after 60 minutes at RT. There was high agreement between EIA and LC/MS-MS. Moreover, we simulated the influence of a 15– or 30-minute delayed incubation on 300 sTXB2 measurements from previously-studied, aspirin-treated patients. Delayed incubation reduced the percentage of aspirin ‘non-responders’ by 22 % to 52 %, depending on the response threshold. In conclusion, a variable delay in the 37 °C incubation of blood samples may affect the assessment of platelet cyclooxygenase-1 inhibition by aspirin and confound the characterization of the determinants of aspirin responsiveness.
High-Resolution Mapping of Postinfarction Reentrant Ventricular Tachycardia
Authors: Anter Elad , Tschabrunn Cory M. , Buxton Alfred E. , Josephson Mark E.
Published: July 2016

Background:

In vivo description of ventricular tachycardia (VT) circuits is limited by insufficient spatiotemporal resolution. We used a novel high-resolution mapping technology to characterize the electrophysiological properties of the postinfarction reentrant VT circuit.

Methods:

In 15 swine, myocardial infarction was induced by left anterior descending artery balloon occlusion. Animals were studied 6 to 8 weeks after myocardial infarction. Activation mapping of VTs was performed by using the Rhythmia mapping system. Activation time was based on a combination of bipolar and unipolar electrograms. The response to overdrive pacing from different zones of the circuit was examined.

Results:

A total of 56 monomorphic VTs were induced (3.8±2.1 per animal). Among these, 21 (37.5%) were hemodynamically stable and allowed mapping of the circuit. Isthmuses were 16.4±7.2 mm long and 7.4±2.8 mm wide. Conduction velocities were slowest at the inward curvature into the isthmus entrance (0.28±0.2 m/s), slightly faster at the outward curvature exit (0.40±0.3 m/s) and nearly normal at the central isthmus (0.62±0.2 m/s). In 3 animals, 2 VT morphologies with opposite axes sharing the same isthmus were mapped. Conduction velocities within the shared isthmus were dependent on the activation vector, consistently slower at the proximal curvature. Overdrive pacing from isthmus sites determined by activation mapping was consistent with entrainment criteria for isthmus. However, dimensions of the isthmus defined by entrainment exceeded dimensions of the isthmus measured by activation mapping by 32±18%.

Conclusions:

In postinfarction reentrant VT, conduction velocities are slowest at the proximal and distal curvatures. Entrainment mapping overestimates the true size of the isthmus. High-resolution activation mapping of VT may better guide ablation therapy.
Defending Shah’s Evidentialism from his Pragmatist Critics: the Carnapian Link
Authors: Hudson Robert
Published: July 2016

In an important 2006 paper, Nishi Shah defends ‘evidentialism’, the position that only evidence for a proposition’s truth constitutes a reason to believe this proposition. In opposition to Shah, Anthony Robert Booth, Andrew Reisner and Asbjørn Steglich-Petersen argue that things other than evidence of truth, so-called non-evidential or ‘pragmatic’ reasons, constitute reasons to believe a proposition. I argue that we can effectively respond to Shah’s pragmatist critics if, following Shah, we are careful to distinguish the evaluation of the reasons for a belief from the process of actually forming a belief and allowing it to influence action. Drawing this distinction is assisted if we utilize Rudolf Carnap’s probabilistic interpretation of what it means to be disposed to believe a claim.
Resurrection of the family Aetobatidae (Myliobatiformes) for the pelagic eagle rays, genus Aetobatus
Authors: WHITE WILLIAM T. , NAYLOR GAVIN J.P.
Published: July 2016

Molecular and morphological data show that the pelagic eagle rays of the genus Aetobatus form a distinct family-level grouping separate from the true eagle rays, Aetomylaeus and Myliobatis (family Myliobatidae). The family Aetobatidae is herein resurrected to include the pelagic eagle rays and definitions are provided for this family and for the Myliobatidae. The key characters separating Aetobatidae from Myliobatidae are: pectoral fins joining head at level of eyes (vs. below level of eyes), internasal flap deeply notched (vs. nearly straight), free rear tip of pectoral fins broadly rounded (vs. angular), spiracles dorsolateral on head and visible in dorsal view (spiracles lateral on head and not visible in dorsal view), dorsal fin with obvious free rear tip (vs. no free rear tip evident, posterior margin joining dorsal surface of tail).