bluuebunny/crossref_metadata_2025 huggingface dataset:
Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration
Authors: Gravel Annie , Dubuc Isabelle , Wallaschek Nina , Gilbert-Girard Shella , Collin Vanessa , Hall-Sedlak Ruth , Jerome Keith R. , Mori Yasuko , Carbonneau Julie , Boivin Guy , Kaufer Benedikt B. , Flamand Louis
Published: May 2017

Human herpesviruses 6A/B (HHV-6A/B) can integrate their viral genomes in the telomeres of human chromosomes. The viral and cellular factors contributing to HHV-6A/B integration remain largely unknown, mostly due to the lack of efficient and reproducible cell culture models to study HHV-6A/B integration. In this study, we characterized the HHV-6A/B integration efficiencies in several human cell lines using two different approaches. First, after a short-term infection (5 h), cells were processed for single-cell cloning and analyzed for chromosomally integrated HHV-6A/B (ciHHV-6A/B). Second, cells were infected with HHV-6A/B and allowed to grow in bulk for 4 weeks or longer and then analyzed for the presence of ciHHV-6. Using quantitative PCR (qPCR), droplet digital PCR, and fluorescent
in situ
hybridization, we could demonstrate that HHV-6A/B integrated in most human cell lines tested, including telomerase-positive (HeLa, MCF-7, HCT-116, and HEK293T) and telomerase-negative cell lines (U2OS and GM847). Our results also indicate that inhibition of DNA replication, using phosphonoacetic acid, did not affect HHV-6A/B integration. Certain clones harboring ciHHV-6A/B spontaneously express viral genes and proteins. Treatment of cells with phorbol ester or histone deacetylase inhibitors triggered the expression of many viral genes, including
U39
,
U90
, and
U100
, without the production of infectious virus, suggesting that the tested stimuli were not sufficient to trigger full reactivation. In summary, both integration models yielded comparable results and should enable the identification of viral and cellular factors contributing to HHV-6A/B integration and the screening of drugs influencing viral gene expression, as well as the release of infectious HHV-6A/B from the integrated state.

IMPORTANCE
The analysis and understanding of HHV-6A/B genome integration into host DNA is currently limited due to the lack of reproducible and efficient viral integration systems. In the present study, we describe two quantitative cell culture viral integration systems. These systems can be used to define cellular and viral factors that play a role in HHV-6A/B integration. Furthermore, these systems will allow us to decipher the conditions resulting in virus gene expression and excision of the integrated viral genome resulting in reactivation.
A case of pulsatile scalp swelling in a child
Authors: Khera Daisy , Khera Pushpinder Singh , Sinha Arvind , Mehta Neeraj
Published: May 2017

Arteriovenous fistulas (AVF) are an entity most commonly seen in the head and neck region. An AVF is a direct connection between the arterial feeders and the draining veins without intervening capillary beds (unlike an arteriovenous malformation in which a nidus intervenes between the arteries and veins). We describe a case of traumatic AVF in a child aged 9 years, who presented with a pulsatile scalp swelling following blunt trauma to the head. It turned out to be an AVF between the superficial temporal artery and vein. Correct diagnosis of an AVF involves Doppler examination with digital subtraction angiography providing a good roadmap for treatment, which can be either endovascular or surgical.
To show the efficacy of compressive sutures alone in the management of acute hydrops in a keratoconus patient
Authors: Subudhi Praveen , Khan Zahiruddin , Subudhi B Nageswar Rao , Sitaram Silla
Published: May 2017

A 17-year-old boy presented with sudden loss of vision in the left eye (OS) for 3 days. He was diagnosed with acute hydrops following keratoconic progression in OS. The patient was initially started on topical medical therapy, including steroids and hypertonic eye drops; showing no signs of resolution. Hence, the patient was planned for full-thickness compressive corneal sutures. Four sutures were placed along the central oedematous area covering the area of ruptured Descemet’s membrane. Signs of resolution were noticed by 1st week and there was complete resolution of oedema by 3rd post-op week. Sutures were removed by 7th postoperative week/45th day. The patient was doing fine with visual acuity of 6/60 in the last visit with no symptoms of discomfort and no signs of corneal vascularisation. Full-thickness corneal compressive sutures is an effective alternative in cases of acute hydrops if perfluoropropane gas is not available.
Full-thickness macular hole: a rare complication ofBorrelia burgdorferineuroretinitis
Authors: Buckle Miranda , Majid Mohammed A , Lee Richard , Steeples Laura R
Published: May 2017

Borrelia burgdorferiis a known infective cause of neuroretinitis. We present a case ofB burgdorferineuroretinitis complicated by macular hole in a 22-year-old man. The neuroretinitis was managed with early high-dose intravenous corticosteroid and oral antibiotic. The macular hole was managed with macular hole surgery after intraocular inflammation had resolved.
Decision making in large animal surgery
Authors: Hallowell Gayle
Published: May 2017

Surgical decision making is a very complex and challenging area of veterinary medicine and this is compounded in farm animal practice by economical constraints. This article discusses some of the conscious and unconscious challenges we have for optimal surgical decision making and considers how we should critique the literature to aid our decision making and improve what we do. Some examples of decision‐making scenarios to help drive individuals' thoughts for their own large animal surgical situations will also be provided.
Scotland's BVD eradication scheme: an update
Authors: Voas S.
Published: May 2017

Scotland's BVD eradication scheme has been running for the past six years and is now in its fourth phase. The chief veterinary officer for Scotland, Sheila Voas, provides an update on where Scotland is now with the scheme and discusses priorities for the future.
The Pricing of Flood Insurance
Authors: Kousky Carolyn , Lingle Brett , Shabman Leonard
Published: May 2017

Despite the availability of flood insurance in the US through the National Flood Insurance Program (NFIP), many homeowners at risk of floods are not insured. The program went deeply into debt after Hurricane Katrina in 2005 and Congress adopted multiple reforms to NFIP rate setting in 2012 and 2014. In response to the ongoing concern about the cost of NFIP policies, as well as multiple misunderstandings about NFIP rates, we simulate NFIP premiums for a sample of homes by constructing a premium calculator based on FEMA rating tables. These simulations highlight several aspects of NFIP rating in need of more attention. First, there are many implicit and explicit cross-subsidies in NFIP pricing, potentially distorting risk information and incentives. Second, NFIP rating is complicated, which can make it difficult for homeowners and untrained agents to evaluate options. Finally, the premiums we present offer insight into the ongoing debate about affordability of flood insurance by highlighting where policies cost the most. FEMA is actively exploring rate reform, which may address some of these concerns.
The Weng'an Biota (Doushantuo Formation): an Ediacaran window on soft-bodied and multicellular microorganisms
Authors: Cunningham John A. , Vargas Kelly , Yin Zongjun , Bengtson Stefan , Donoghue Philip C. J.
Published: May 2017

The Weng'an Biota is a fossil Konservat-Lagerstätte in South China that is
c.
570 – 609 myr old and provides an unparalleled snapshot of marine life during the interval in which molecular clocks estimate that animal clades were diversifying. It yields fossils that are three-dimensionally preserved in calcium phosphate with cellular and sometimes subcellular fidelity. The biota includes candidates for the oldest animals in the fossil record, including embryonic, larval and adult forms. We argue that, although the Weng'an Biota includes forms that could be animals, none can currently be assigned to this group with confidence. Nonetheless, the biota offers a rare and valuable window on the evolution of multicellular and soft-bodied organisms in the prelude to the Cambrian radiation.
The Evolution of Organellar Coat Complexes and Organization of the Eukaryotic Cell
Authors: Rout Michael P. , Field Mark C.
Published: May 2017

Eukaryotic cells possess a remarkably diverse range of organelles that provide compartmentalization for distinct cellular functions and are likely responsible for the remarkable success of these organisms. The origins and subsequent elaboration of these compartments represent a key aspect in the transition between prokaryotic and eukaryotic cellular forms. The protein machinery required to build, maintain, and define many membrane-bound compartments is encoded by several paralog families, including small GTPases, coiled-bundle proteins, and proteins with β-propeller and α-solenoid secondary structures. Together these proteins provide the membrane coats and control systems to structure and coordinate the endomembrane system. Mechanistically and evolutionarily, they unite not only secretory and endocytic organelles but also the flagellum and nucleus. The ancient origins for these families have been revealed by recent findings, providing new perspectives on the deep evolutionary processes and relationships that underlie eukaryotic cell structure.
A novel role for primary cilia in airway remodeling
Authors: Trempus Carol S. , Song Weifeng , Lazrak Ahmed , Yu Zhihong , Creighton Judy R. , Young Bethany M. , Heise Rebecca L. , Yu Yen Rei , Ingram Jennifer L. , Tighe Robert M. , Matalon Sadis , Garantziotis Stavros
Published: May 2017

Primary cilia (PC) are solitary cellular organelles that play critical roles in development, homeostasis, and disease pathogenesis by modulating key signaling pathways such as Sonic Hedgehog and calcium flux. The antenna-like shape of PC enables them also to facilitate sensing of extracellular and mechanical stimuli into the cell, and a critical role for PC has been described for mesenchymal cells such as chondrocytes. However, nothing is known about the role of PC in airway smooth muscle cells (ASMCs) in the context of airway remodeling. We hypothesized that PC on ASMCs mediate cell contraction and are thus integral in the remodeling process. We found that PC are expressed on ASMCs in asthmatic lungs. Using pharmacological and genetic methods, we demonstrated that PC are necessary for ASMC contraction in a collagen gel three-dimensional model both in the absence of external stimulus and in response to the extracellular component hyaluronan. Mechanistically, we demonstrate that the effect of PC on ASMC contraction is, to a small extent, due to their effect on Sonic Hedgehog signaling and, to a larger extent, due to their effect on calcium influx and membrane depolarization. In conclusion, PC are necessary for the development of airway remodeling by mediating calcium flux and Sonic Hedgehog signaling.
Regulation of axonal regeneration following spinal cord injury in the lamprey
Authors: Benes Jessica A. , House Kylie N. , Burks Frank N. , Conaway Kris P. , Julien Donald P. , Donley Jeffrey P. , Iyamu Michael A. , McClellan Andrew D.
Published: May 2017

Following rostral spinal cord injury (SCI) in larval lampreys, injured descending brain neurons, particularly reticulospinal (RS) neurons, regenerate their axons, and locomotor behavior recovers in a few weeks. However, axonal regeneration of descending brain neurons is mostly limited to relatively short distances, but the mechanisms for incomplete axonal regeneration are unclear. First, lampreys with rostral SCI exhibited greater axonal regeneration of descending brain neurons, including RS neurons, as well as more rapid recovery of locomotor muscle activity right below the lesion site, compared with animals with caudal SCI. In addition, following rostral SCI, most injured RS neurons displayed the “injury phenotype,” whereas following caudal SCI, most injured neurons displayed normal electrical properties. Second, following rostral SCI, at cold temperatures (~4–5°C), axonal transport was suppressed, axonal regeneration and behavioral recovery were blocked, and injured RS neurons displayed normal electrical properties. Cold temperatures appear to prevent injured RS neurons from detecting and/or responding to SCI. It is hypothesized that following rostral SCI, injured descending brain neurons are strongly stimulated to regenerate their axons, presumably because of elimination of spinal synapses and reduced neurotrophic support. However, when these neurons regenerate their axons and make synapses right below the lesion site, restoration of neurotrophic support very likely suppress further axonal regeneration. In contrast, caudal SCI is a weak stimulus for axonal regeneration, presumably because of spared synapses above the lesion site. These results may have implications for mammalian SCI, which can spare synapses above the lesion site for supraspinal descending neurons and propriospinal neurons.

NEW & NOTEWORTHY Lampreys with rostral spinal cord injury (SCI) exhibited greater axonal regeneration of descending brain neurons and more rapid recovery of locomotor muscle activity below the lesion site compared with animals with caudal SCI. In addition, following rostral SCI, most injured reticulospinal (RS) neurons displayed the “injury phenotype,” whereas following caudal SCI, most injured neurons had normal electrical properties. We hypothesize that following caudal SCI, the spared synapses of injured RS neurons might limit axonal regeneration and behavioral recovery.
Dental Enamel Formation and Implications for Oral Health and Disease
Authors: Lacruz Rodrigo S. , Habelitz Stefan , Wright J. Timothy , Paine Michael L.
Published: May 2017

Dental enamel is the hardest and most mineralized tissue in extinct and extant vertebrate species and provides maximum durability that allows teeth to function as weapons and/or tools as well as for food processing. Enamel development and mineralization is an intricate process tightly regulated by cells of the enamel organ called ameloblasts. These heavily polarized cells form a monolayer around the developing enamel tissue and move as a single forming front in specified directions as they lay down a proteinaceous matrix that serves as a template for crystal growth. Ameloblasts maintain intercellular connections creating a semi-permeable barrier that at one end (basal/proximal) receives nutrients and ions from blood vessels, and at the opposite end (secretory/apical/distal) forms extracellular crystals within specified pH conditions. In this unique environment, ameloblasts orchestrate crystal growth via multiple cellular activities including modulating the transport of minerals and ions, pH regulation, proteolysis, and endocytosis. In many vertebrates, the bulk of the enamel tissue volume is first formed and subsequently mineralized by these same cells as they retransform their morphology and function. Cell death by apoptosis and regression are the fates of many ameloblasts following enamel maturation, and what cells remain of the enamel organ are shed during tooth eruption, or are incorporated into the tooth’s epithelial attachment to the oral gingiva. In this review, we examine key aspects of dental enamel formation, from its developmental genesis to the ever-increasing wealth of data on the mechanisms mediating ionic transport, as well as the clinical outcomes resulting from abnormal ameloblast function.
Method for Solving LASSO Problem Based on Multidimensional Weight
Authors: ChunRong Chen , ShanXiong Chen , Lin Chen , YuChen Zhu
Published: May 2017

In the data mining, the analysis of high-dimensional data is a critical but thorny research topic. The LASSO (least absolute shrinkage and selection operator) algorithm avoids the limitations, which generally employ stepwise regression with information criteria to choose the optimal model, existing in traditional methods. The improved-LARS (Least Angle Regression) algorithm solves the LASSO effectively. This paper presents an improved-LARS algorithm, which is constructed on the basis of multidimensional weight and intends to solve the problems in LASSO. Specifically, in order to distinguish the impact of each variable in the regression, we have separately introduced part of principal component analysis (Part_PCA), Independent Weight evaluation, and CRITIC, into our proposal. We have explored that these methods supported by our proposal change the regression track by weighted every individual, to optimize the approach direction, as well as the approach variable selection. As a consequence, our proposed algorithm can yield better results in the promise direction. Furthermore, we have illustrated the excellent property of LARS algorithm based on multidimensional weight by the Pima Indians Diabetes. The experiment results show an attractive performance improvement resulting from the proposed method, compared with the improved-LARS, when they are subjected to the same threshold value.
Circularly Polarized Antenna Array Fed by Air-Bridge Free CPW-Slotline Network
Authors: Liu Yilin , Huang Kama , Luo Xia
Published: May 2017

A novel design of 1×2 and 2×2 circularly polarized (CP) microstrip patch antenna arrays is presented in this paper. The two CP antenna arrays are fed by sequentially rotated coplanar waveguide (CPW) to slotline networks and are processed on 1 mm thick single-layer FR4 substrates. Both of the two arrays are low-profile and lightweight. An air-bridge free CPW-slotline power splitter is appropriately designed to form the feeding networks and realize the two CP antenna arrays. The mechanism of circular polarization in this design is explained. The simulated and measured impedance bandwidths as well as the 3 dB axial ratio bandwidths and the radiation patterns of the two proposed antenna arrays are presented. This proposed design can be easily extended to form a larger plane array with good performance owing to its simple structure.
Multitiered and Distributed WSAN for Cooperative Indoors Environment Management
Authors: Heo Taewook , Kim Hyunhak , Doh Yoonmee , Kim Kwangsoo , Park Jongjun , Kim Naesoo , Jun JongArm , Ko JeongGil
Published: May 2017

For the past decade, wireless sensor networks have focused primarily on data collection. As a result the network topology for these systems was usually heavily centralized. However, for these networks to form a full system, the introduction of proper actuation units and decision-making intelligence is inevitable. Such a new wireless sensor and actuator network system enables new architectural research issues that have not been previously studied. In this work, we introduce the DWSAN system architecture, which effectively combines both sensor and actuation hardware devices to a single network and manages this network so that the actuation decisions are made in a distributed manner and the topology of the network maintains a multitier architecture. Our intensive set of evaluations reveal that, compared to the centralized approach that has been used in most wireless sensor network systems until now, when actuation units are introduced to the system, the DWSAN architecture reduces the transmission load of the network and the actuation decision-making latency by close to twofold and threefold, respectively. Furthermore, we show that this benefit naturally leads to better scalability of the system, making it suitable for various sensing applications in different environments.
New Mathematical Model for the Surface Area of the Left Ventricle by the Truncated Prolate Spheroid
Authors: Domingues José Sérgio , Vale Marcos de Paula , Martinez Carlos Barreira
Published: May 2017

The main aim of this study was the formula application of the superficial area of a truncated prolate spheroid (TPS) in Cartesian coordinates in obtaining a cardiac parameter that is not so much discussed in literature, related to the left ventricle (LV) surface area of the human heart, by age and sex. First we obtain a formula for the area of a TPS. Then a simple mathematical model of association of the axes measures of a TPS with the axes of the LV is built. Finally real values of the average dimensions of the humans LV are used to measure surface areas approximations of this heart chamber. As a result, the average superficial area of LV for normal patients is obtained and it is observed that the percentage differences of areas between men and women and their consecutive age groups are constant. A strong linear correlation between the obtained areas and the ventricular volumes normalized by the body areas was observed. The obtained results indicate that the superficial area of the LV, besides enabling a greater knowledge of the geometrical characteristics of the human LV, may be used as one of the normality cardiac verification criteria and be useful for medical and biological applications.
Organic Dyes Containing Coplanar Dihexyl-Substituted Dithienosilole Groups for Efficient Dye-Sensitised Solar Cells
Authors: Lyons Ciaran , Rathi Neelima , Dev Pratibha , Byrne Owen , Surolia Praveen K. , Maji Pathik , MacElroy J. M. D. , Yella Aswani , Grätzel Michael , Magner Edmond , English Niall J. , Thampi K. Ravindranathan
Published: May 2017

A chromophore containing a coplanar dihexyl-substituted dithienosilole (CL1) synthesised for use in dye-sensitised solar cells displayed an energy conversion efficiency of 6.90% under AM 1.5 sunlight irradiation. The new sensitiser showed a similar fill factor and open-circuit voltage when compared with N719. Impedance measurements showed that, in the dark, the charge-transfer resistance of a cell using CL1 in the intermediate-frequency region was higher compared to N719 (69.8 versus 41.3 Ω). Under illumination at AM 1.5G-simulated conditions, the charge-transfer resistances were comparable, indicative of similar recombination rates by the oxidised form of the redox couple. The dye showed instability in ethanol solution, but excellent stability when attached to TiO2. Classical molecular dynamics indicated that interactions between ethanol and the dye are likely to reduce the stability of CL1 in solution form. Time-dependent density functional theory studies were performed to ascertain the absorption spectrum of the dye and assess the contribution of various transitions to optical excitation, which showed good agreement with experimental results.
Determining the Coupling Source on a Set of Oscillators from Experimental Data
Authors: Jauregui-Correa Juan Carlos , López-Cajun Carlos S. , Sen Mihir
Published: May 2017

Complex systems are a broad concept that comprises many disciplines, including engineering systems. Regardless of their particular behavior, complex systems share similar behaviors, such as synchronization. This paper presents different techniques for determining the source of coupling when a set of oscillators synchronize. It is possible to identify the location and time variations of the coupling by applying a combination of analytical techniques, namely, the source of synchronization. For this purpose, the analysis of experimental data from a complex mechanical system is presented. The experiment consisted in placing a 24-bladed rotor under an airflow. The vibratory motion of the blades was recorded with accelerometers, and the resulting information was analyzed with four techniques: correlation coefficients, Kuramoto parameter, cross-correlation functions, and the recurrence plot. The measurements clearly show the existence of frequencies due to the foreground components and the internal interaction between them due to the background components (coupling).
Phase I Trial of Intratumoral Injection ofCCL21Gene–Modified Dendritic Cells in Lung Cancer Elicits Tumor-Specific Immune Responses and CD8+ T-cell Infiltration
Authors: Lee Jay M. , Lee Mi-Heon , Garon Edward , Goldman Jonathan W. , Salehi-Rad Ramin , Baratelli Felicita E. , Schaue Dörthe , Wang Gerald , Rosen Fran , Yanagawa Jane , Walser Tonya C. , Lin Ying , Park Stacy J. , Adams Sharon , Marincola Francesco M. , Tumeh Paul C. , Abtin Fereidoun , Suh Robert , Reckamp Karen L. , Lee Gina , Wallace William D. , Lee Sarah , Zeng Gang , Elashoff David A. , Sharma Sherven , Dubinett Steven M.
Published: May 2017

Purpose: A phase I study was conducted to determine safety, clinical efficacy, and antitumor immune responses in patients with advanced non–small cell lung carcinoma (NSCLC) following intratumoral administration of autologous dendritic cells (DC) transduced with an adenoviral (Ad) vector expressing the CCL21 gene (Ad-CCL21-DC). We evaluated safety and tumor antigen–specific immune responses following in situ vaccination (ClinicalTrials.gov: NCT01574222).

Experimental Design: Sixteen stage IIIB/IV NSCLC subjects received two vaccinations (1 × 106, 5 × 106, 1 × 107, or 3 × 107 DCs/injection) by CT- or bronchoscopic-guided intratumoral injections (days 0 and 7). Immune responses were assessed by tumor antigen–specific peripheral blood lymphocyte induction of IFNγ in ELISPOT assays. Tumor biopsies were evaluated for CD8+ T cells by IHC and for PD-L1 expression by IHC and real-time PCR (RT-PCR).

Results: Twenty-five percent (4/16) of patients had stable disease at day 56. Median survival was 3.9 months. ELISPOT assays revealed 6 of 16 patients had systemic responses against tumor-associated antigens (TAA). Tumor CD8+ T-cell infiltration was induced in 54% of subjects (7/13; 3.4-fold average increase in the number of CD8+ T cells per mm2). Patients with increased CD8+ T cells following vaccination showed significantly increased PD-L1 mRNA expression.

Conclusions: Intratumoral vaccination with Ad-CCL21-DC resulted in (i) induction of systemic tumor antigen–specific immune responses; (ii) enhanced tumor CD8+ T-cell infiltration; and (iii) increased tumor PD-L1 expression. Future studies will evaluate the role of combination therapies with PD-1/PD-L1 checkpoint inhibition combined with DC-CCL21 in situ vaccination. Clin Cancer Res; 23(16); 4556–68. ©2017 AACR.
The “Tricky Business” of Identifying Mechanisms of Resistance to Anti–PD-1
Authors: Wilmott James S. , Rizos Helen , Scolyer Richard A. , Long Georgina V.
Published: May 2017

Resistance to immune checkpoint inhibitors can vary between patients and among metastases. Understanding the genomic, transcriptomic, and microenvironmental factors that contribute to this variability will reveal the mechanisms that tumors utilize to evade the therapeutic effects of checkpoint inhibitor immunotherapies and will enable us to develop strategies to overcome them. Clin Cancer Res; 23(12); 2921–3. ©2017 AACR.

See related article by Ascierto et al., p. 3168
Vulnerability of Small-Cell Lung Cancer to Apoptosis Induced by the Combination of BET Bromodomain Proteins and BCL2 Inhibitors
Authors: Lam Lloyd T. , Lin Xiaoyu , Faivre Emily J. , Yang Ziping , Huang Xiaoli , Wilcox Denise M. , Bellin Richard J. , Jin Sha , Tahir Stephen K. , Mitten Michael , Magoc Terry , Bhathena Anahita , Kati Warren M. , Albert Daniel H. , Shen Yu , Uziel Tamar
Published: May 2017

Ten percent to 15% of all lung cancers are small-cell lung cancer (SCLC). SCLC usually grows and metastasizes before it is diagnosed and relapses rapidly upon treatment. Unfortunately, no new targeted agent has been approved in the past 30 years for patients with SCLC. The BET (bromodomain and extraterminal) proteins bind acetylated histones and recruit protein complexes to promote transcription initiation and elongation. BET proteins have been shown to regulate expression of key genes in oncogenesis, such as MYC, CCND2, and BCL2L1. Here, we demonstrate that approximately 50% of SCLC cell lines are exquisitely sensitive to growth inhibition by the BET inhibitor, ABBV-075. The majority of these SCLC cell lines underwent apoptosis in response to ABBV-075 treatment via induction of caspase-3/7 activity. ABBV-075 enhanced the expression of proapoptotic protein BIM and downregulated antiapoptotic proteins BCL2 and BCLxl to a lesser extent. Furthermore, BET inhibition increased BCL2–BIM complex, thus priming the cells for apoptosis. Indeed, strong synergy was observed both in vitro and in vivo when cotreating the cells with BET inhibitor and the BH3-mimetic, BCL2 inhibitor venetoclax (ABT-199). ABBV-075 interaction with venetoclax positively correlated with BCL2 expression. Taken together, our studies provide a rationale for treating SCLC with BET and BCL2 inhibitors in tumors with high BCL2 protein expression. Mol Cancer Ther; 16(8); 1511–20. ©2017 AACR.
Synergy between Androgen Receptor Antagonism and Inhibition of mTOR and HER2 in Breast Cancer
Authors: Gordon Michael A. , D'Amato Nicholas C. , Gu Haihua , Babbs Beatrice , Wulfkuhle Julia , Petricoin Emanuel F. , Gallagher Isela , Dong Ting , Torkko Kathleen , Liu Bolin , Elias Anthony , Richer Jennifer K.
Published: May 2017

The androgen receptor (AR) is widely expressed in breast cancer, and evidence suggests dependence on AR signaling for growth and survival. AR antagonists such as enzalutamide and seviteronel have shown success in preclinical models and clinical trials of prostate cancer and are currently being evaluated in breast cancer. Reciprocal regulation between AR and the HER2/PI3K/mTOR pathway may contribute to resistance to HER2- and mTOR-targeted therapies; thus, dual inhibition of these pathways may synergistically inhibit breast cancer growth. HER2+ and triple-negative breast cancer cell lines were treated with AR antagonist plus anti-HER2 mAb trastuzumab or mTOR inhibitor everolimus. Apoptosis, cell proliferation, and drug synergy were measured in vitro. Pathway component genes and proteins were measured by qRT-PCR, Western blot, and reverse phase protein array. In vivo, HER2+ breast cancer xenografts were treated with enzalutamide, everolimus, trastuzumab, and combinations of these drugs. AR antagonists inhibited proliferation of both HER2+ and TNBC cell lines. Combining AR antagonist and either everolimus or trastuzumab resulted in synergistic inhibition of proliferation. Dihydrotestosterone caused increased phosphorylation of HER2 and/or HER3 that was attenuated by AR inhibition. Everolimus caused an increase in total AR, phosphorylation of HER2 and/or HER3, and these effects were abrogated by enzalutamide. Growth of trastuzumab-resistant HER2+ xenograft tumors was inhibited by enzalutamide, and combining enzalutamide with everolimus decreased tumor viability more than either single agent. AR antagonists synergize with FDA-approved breast cancer therapies such as everolimus and trastuzumab through distinct mechanisms. Treatment combinations are effective in trastuzumab-resistant HER2+ breast cancer cells in vivo. Mol Cancer Ther; 16(7); 1389–400. ©2017 AACR.
Combining DNA Vaccine and AIDA-1 in AttenuatedSalmonellaActivates Tumor-Specific CD4+ and CD8+ T-cell Responses
Authors: Mei Yu , Zhao Lixiang , Liu Yonghao , Gong Huanle , Song Yuan , Lei Lei , Zhu Ying , Jin Ziqi , Ma Shoubao , Hu Bo , Sun Qing , Liu Haiyan
Published: May 2017

Stimulation of tumor-specific responses in both CD4+ and CD8+ T cells has been a challenge for effective tumor vaccines. We designed a vaccine vector containing the AIDA-1 autotransporter and DNA vaccine elements, generating a murine melanoma vaccine that was delivered by the attenuated Salmonella strain SL7207. Growth of murine subcutaneous melanoma was significantly inhibited by intranasal immunization with the Salmonella tumor vaccine. The vaccine activated tumor-specific CD4+ and CD8+ T-cell responses, with increased T-cell proliferation, tumor antigen–specific Th1 cytokine production, increased percentages of tetramer positive cells, and cytotoxicity. CD4+ or CD8+ T-cell depletion resulted in the loss of antitumor activity of the Salmonella tumor vaccine, suggesting that the efficacy of the vaccine was dependent on both CD4+ and CD8+ T cells. Lung metastasis of the tumor was also inhibited by vaccine treatment. Similarly, the percentages of tumor-specific Th1 cytokine production by CD4+ and CD8+ T cells in the spleen, tumor, and bronchoalveolar lavage were increased after vaccine treatment. Tumor-specific proliferation of CD4+ and CD8+ T cells was also promoted by the vaccine. Tetramer staining and cytotoxicity assay showed enhanced tumor-specific CD8+ T-cell response after vaccine treatment. Therefore, the Salmonella tumor vaccine could activate both tumor-specific CD4+ and CD8+ T-cell responses. This vaccine strategy may be widely applicable to the development of oral or nasal vaccines against tumors. Cancer Immunol Res; 5(6); 503–14. ©2017 AACR.
WNT5A Correlates with Clinicopathological Characteristics in Gastric Cancer: a Meta-Analysis
Authors: Nam Seungyoon , Chung Jun-Won , Yang Jun-Young
Published: May 2017

Background/Aims: Gastric cancer (GC), the third-leading cause of cancer death in the world, is typically diagnosed only in its advanced stages. WNT signaling has been associated with clinicopathological characteristics in diverse cancer types. But the systematic analysis of WNT5A, a member in the signaling, has not been inspected. Thus, our study used a meta-analysis to statistically associate WNT5A expression with GC clinicopathological characteristics. Methods: For a systematic literature review of GC in combination with the WNT signaling molecule WNT5A, we searched for PubMed, Cochrane Library, and Web of Science. It led to the five cohorts, in four eligible studies, consisting of 1,034 patients (617 WNT5A-positive and 417 WNT5A-negative patients). These patients were inspected by the library “meta” in R software for our meta-analysis. Results: Our meta-analysis, revealed a statistically significant associations of WNT5A-positivity with lymph node metastasis (p=0.0047), some types of Lauren diffuse subtype GCs (p<0.0001), advanced tumor depth (p<0.0001), and advanced UICC stages (p=0.0461) with no observation of bias or confounding factors. Conclusions: These results support the feasibility of targeting this embryonic signaling pathway, both for therapy, and as a biomarker to “guide” various individual interventions (i.e., “personalized medicine”).
Hepatitis B Virus Core Antigen Stimulates IL-6 Expression via p38, ERK and NF-κB Pathways in Hepatocytes
Authors: Chen Zhi , Li Yang-Xia , Fu Hai-Jing , Ren Yan-Li , Zou Ling , Shen Shi-Zhen , Chen Ping , Sun Ting , Huang Chun-Hong
Published: May 2017

Background: Hepatitis B virus (HBV) causes both acute and chronic liver injury. Viral proteins are involved in the pathological progress. Hepatitis B core antigen (HBcAg), a component of viral nucleocapsid, is not only essential for HBV lifecycle, but also exhibits strong immunogenicity. The cytoplasmic location of HBcAg in liver biopsy is associated with liver injury and inflammation, but the exact mechanisms remain to be elaborated. Methods: Huh7, SMMC-7721 and L-02 cells were transfected with pEGFP-N1-HBcAg to establish an intracellular HBcAg expression model. The mRNA and protein levels of Interleukin (IL)-6 were detected by qPCR and ELISA respectively. The signaling pathway-related proteins were investigated by western blot and immunofluorescence assay. Results: HBcAg increased the expression and secretion of IL-6 through activating extracellular signal-related kinase (ERK), p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor-kappa B (NF-κB). These activations can be blocked by specific inhibitors of the three pathways. Conclusions: HBcAg actives p38, ERK1/2 and NF-κB to enhance the production of IL-6 in hepatocytes. This provides a molecular mechanism to explain the association of cytoplasmic HBcAg with severe liver injury and inflammation.
MiR-21 Regulates TNF-α-Induced CD40 Expression via the SIRT1-NF-κB Pathway in Renal Inner Medullary Collecting Duct Cells
Authors: Lin Qinqin , Geng Yuanwen , Zhao Meng , Lin Shuaishuai , Zhu Qing , Tian Zhenjun
Published: May 2017

Background/Aims: Recent studies have indicated that microRNA-21 (miR-21) is involved in the inflammatory response in relation to renal disease. Sirtuin1 (SIRT1) exerts renoprotective properties by counteracting inflammation. The activation of CD40 triggers inflammation that participates in renal inflammation and injury. The relationship between miR-21, SIRT1 and CD40, however, remains elusive. Methods: Immunohistochemistry, small-interfering RNA (siRNA) transfection, quantitative real-time PCR and western blotting were applied to assess the morphological, functional and molecular mechanisms in primary cultured renal inner medullary collecting duct (IMCD) cells. Results: TNF-α induced miR-21, CD40 and acetylated-NF-κBp65 (Ac-p65) expressions and reduced SIRT1 expression in IMCD cells. miR-21 mimics increased SIRT1 expression and attenuated Ac-p65 and CD40 expressions in TNF-α-induced IMCD cells, and the corresponding changes were observed with a miR-21 inhibitor. SIRT1 overexpression or activation by SRT1720 diminished TNF-α-induced CD40 and Ac-p65 expressions, which was reversed by SIRT1 siRNA or the inhibitors Ex527 and sirtinol and augmented by pretreatment with NF-κB siRNA. Further study found that the inhibitory effect of miR-21 on Ac-p65 and CD40 expressions was impeded by pretreatment with SIRT1 siRNA. Conclusion: The present study indicates that miR-21 inhibits TNF-α-induced CD40 expression in IMCD cells via the SIRT1-NF-κB signalling pathway, which provides new insight in understanding the anti-inflammatory effect of miR-21.
Role for Target of Rapamycin (mTOR) Signal Pathway in Regulating Neuronal Injury after Intracerebral Hemorrhage
Authors: Wang Jie-Ping , Zhang Meng-Yu
Published: May 2017

Background/Aims: Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase and activation of its signal pathway plays an important role in regulating protein growth and synthesis as well as cell proliferation and survival. In the present study, we examined the contribution of mTOR signal and its downstream products to brain injuries induced by intracerebral hemorrhage (ICH) in rats. Methods: Western Blot analysis was employed to examine the protein expression of mTOR and its downstream pathway and Caspase-3. ELISA was used to measure pro-inflammatory cytokines (PICs) and vascular endothelial growth factor (VEGF). Additionally, neurological Severity Score and brain water content were used to indicate neurological function and brain edema. Results: The protein expression of p-mTOR, mTOR-mediated phosphorylation of 4E–binding protein 4 (4E-BP1), p70 ribosomal S6 protein kinase 1 (S6K1) pathways were amplified in ICH rats compared with controls. Blocking mTOR using rapamycin significantly attenuated upregulation of PICs, namely IL-1β, IL-6 and TNF-α, and Caspase-3 indicating cell apoptosis, and promoted the levels of VEGF and its subtype receptor VEGFR-2 in brain tissues. Moreover, the effects of rapamycin were linked to improvement of neurological deficits and increased brain water content observed in ICH rats. Conclusion: Activation mTOR signal is engaged in pathophysiological process during ICH and blocking mTOR pathway plays a beneficial role in regulating neuronal tissues via PIC, apoptotic Caspase-3 and VEGF mechanisms. This has pharmacological implications to target specific mTOR and its downstream signal pathway for neuronal dysfunction and vulnerability related to ICH.
Three-Year Experience in NAT Screening of Blood Donors for Transfusion Transmitted Viruses in Croatia
Authors: Safic Stanic Hana , Babic Ivana , Maslovic Margareta , Dogic Vesna , Bingulac-Popovic Jasna , Miletic Manuela , Jurakovic-Loncar Nina , Vuk Tomislav , Strauss-Patko Maja , Jukic Irena
Published: May 2017

Background: Croatia implemented individual donation (ID)-NAT testing of blood donors in 2013 for three viruses HBV, HCV, and HIV-1 as a mandatory test for all blood donors. This study assessed the impact of NAT screening 3 years after its implementation. Methods: A total of 545,463 donations were collected and screened for HBV, HCV, and HIV-1 using the Procleix Ultrio Plus Assay. All initially reactive (IR) NAT samples were retested in triplicate and, if repeatedly reactive (RR), NAT discriminatory assay (dNAT) was performed. ID-NAT positive donations were confirmed by RT-PCR on the COBAS AmpliPrep/TaqMan platform. Results: Out of 545,463 samples tested, 108 (0.02%) were RR in NAT. There were 82 (75,9%) HBV reactive, 16 (14.8%) HCV reactive, and 10 (9.3%) HIV-1 reactive samples. 51 (47.2%) samples were ID-NAT positive only. Out of these 51 NAT yield cases, 1 window period HIV-1 and 50 occult HBV infections (OBI) were determined. There were only two potential HBV DNA transmissions from OBI donors. Conclusion: The implementation of NAT screening for three viruses has improved blood safety in Croatia. During the 3-year period, 1 window period HIV-1 and a number of occult HBV donations were identified.
Role of Organic Anion Transporters in the Uptake of Protein-Bound Uremic Toxins by Human Endothelial Cells and Monocyte Chemoattractant Protein-1 Expression
Authors: Favretto Giane , Souza Lauro M. , Gregório Paulo C. , Cunha Regiane S. , Maciel Rayana A.P. , Sassaki Guilherme L. , Toledo Maria G. , Pecoits-Filho Roberto , Souza Wesley M. , Stinghen Andréa E.M.
Published: May 2017

Organic anion transporters (OATs) are involved in the uptake of uremic toxins such as p-cresyl sulfate (PCS) and indoxyl sulfate (IS), which play a role in endothelial dysfunction in patients with chronic kidney diseases (CKD). In this study, we investigated the role of OAT1 and OAT3 in the uptake of PCS and IS into human endothelial cells. PCS was synthesized via p-cresol sulfation and characterized using analytical methods. The cells were treated with PCS and IS in the absence and presence of probenecid (Pb), an OAT inhibitor. Cell viability was assessed using the MTT assay. The absorbed toxins were analyzed using chromatography, OAT expression using immunocytochemistry and western blot, and monocyte chemoattractant protein-1 (MCP-1) expression using enzyme-linked immunosorbent assay. Cell viability decreased after toxin treatment in a dose-dependent manner. PCS and IS showed significant internalization after 60 min treatment, while no internalization was observed in the presence of Pb, suggesting that OATs are involved in the transport of both toxins. Immunocytochemistry and western blot demonstrated OAT1 and OAT3 expression in endothelial cells. MCP-1 expression increased after toxins treatment but decreased after Pb treatment. PCS and IS uptake were mediated by OATs, and OAT blockage could serve as a therapeutic strategy to inhibit MCP-1 expression.
Agreement between Estimated and Measured Renal Function in an Everyday Clinical Outpatient Setting of Human Immunodeficiency Virus-Infected Individuals
Authors: Ahlström Magnus Glindvad , Kjær Andreas , Gerstoft Jan , Obel Niels
Published: May 2017

Introduction: Estimated renal function (eRF) has been widely implemented as a screening tool in handling human immunodeficiency virus (HIV)-infected individuals. Our primary objective was to investigate the agreement between measured renal function (mRF) and eRF in HIV-infected individuals in an everyday clinical setting. Methods: A single-center study at the HIV-outpatient clinic at Copenhagen University Hospital, Rigshospitalet. Study period from January 1, 2004-June 1, 2015. We included all HIV-infected individuals who had an mRF performed and compared this with eRF assessed with 9 different serum-creatinine-based equations and the eRF reported by the Department of Clinical Biochemistry. We evaluated performance characteristics of the different eRFs, with concordance correlation coefficient, total deviation index, coverage probability, relative accuracy, and Bland Altman plots. We also evaluated whether exposure to (1) rilpivirine, cobicistat, or dolutegravir (RLP/COB/DTG), (2) protease inhibitors (PIs), or (3) tenofovir disoproxil fumarate (TDF) had an impact on agreement. Furthermore, we compared inter- and intra-individual differences between mRF and eRF. Results: Ninety-eight individuals had an mRF performed during the study period. We found that the agreement between mRF and eRF was poor irrespective of the eRF equation. Exposure to RLP/COB/DTG and PIs was not associated with different agreement. Exposure to TDF was associated with statistically significant better agreement for 3 of the evaluated equations. Conclusion: Irrespective of calculation methods, the agreement between mRF and eRF is poor. Surprisingly TDF exposure was associated with a better agreement compared with TDF-unexposed individuals.
St. Gallen/Vienna 2017: A Brief Summary of the Consensus Discussion about Escalation and De-Escalation of Primary Breast Cancer Treatment
Authors: Gnant Michael , Harbeck Nadia , Thomssen Christoph
Published: May 2017

For the second time, the St. Gallen Consensus Conference on early breast cancer treatment standards took place in Vienna, Austria, where it will remain for the foreseeable future (next date: March 20-23, 2019). With the probably most prominent line-up of global breast cancer experts and more than 3,000 participants from over 100 countries, the 2017 St. Gallen/Vienna conference again was a huge success. A generation change took place with respect to the Conference Co-Chairpersons. Traditionally, the experts from all continents reviewed publications from the past 2 years, and discussed whether new diagnostic or therapeutic means were ready for routine everyday practice. This year, the conference's main theme was ‘Escalating and Deescalating Treatment', and the traditional panel votings clarified a number of issues in this respect. Several subjects of all breast cancer modalities were further de-escalated (surgery: ‘no ink on tumor' clearly confirmed as standard; resection within new limits after neoadjuvant systemic therapy; axillary dissection may also be avoided after mastectomy under certain circumstances; radiotherapy: hypofractionation is standard of care in breast conserving therapy; chemotherapy: can be avoided in low-risk patients). However, others were escalated: surgery: after neoadjuvant treatment and after mastectomy a positive sentinel node leads to axillary dissection; radiotherapy: regional nodes have to be irradiated in 4+ nodes situations; adjuvant therapy: bisphosphonates as standard for postmenopausal women. There was no clear panel opinion on the optimal use of multigenomic assays. As always, the panel recommendations are strictly opinion-based, and try to depict the ‘usual' treatment for the ‘average' patients. This rapid report by the editors-in-chief of Breast Care summarizes the results of the 2017 international panel votings with respect to loco-regional systemic treatment, and does not intend to replace the official St. Gallen Consensus publication.