bluuebunny/crossref_metadata_2025 huggingface dataset:
Vibrio harveyi virulence gene expression in vitro and in vivo during infection in black tiger shrimp Penaeus monodon
Authors: Peeralil S , Joseph TC , Murugadas V , Akhilnath PG , Sreejith VN , Lalitha KV
Published: April 2020

Luminescent Vibrio harveyi is common in sea and estuarine waters. It produces several virulence factors and negatively affects larval penaeid shrimp in hatcheries, resulting in severe economic losses to shrimp aquaculture. Although V. harveyi is an important pathogen of shrimp, its pathogenicity mechanisms have yet to be completely elucidated. In the present study, isolates of V. harveyi were isolated and characterized from diseased Penaeus monodon postlarvae from hatcheries in Kerala, India, from September to December 2016. All 23 tested isolates were positive for lipase, phospholipase, caseinase, gelatinase and chitinase activity, and 3 of the isolates (MFB32, MFB71 and MFB68) showed potential for significant biofilm formation. Based on the presence of virulence genes, the isolates of V. harveyi were grouped into 6 genotypes, predominated by vhpA+ flaB+ ser+ vhh1- luxR+ vopD- vcrD+ vscN-. One isolate from each genotype was randomly selected for in vivo virulence experiments, and the LD50 ranged from 1.7 ± 0.5 × 103 to 4.1 ± 0.1 × 105 CFU ml-1. The expression of genes during the infection in postlarvae was high in 2 of the isolates (MFB12 and MFB32), consistent with the result of the challenge test. However, in MFB19, even though all genes tested were present, their expression level was very low and likely contributed to its lack of virulence. Because of the significant variation in gene expression, the presence of virulence genes alone cannot be used as a marker for pathogenicity of V. harveyi.
Scotland's Lost Colony Found: Rediscovering Stuarts Town, 1682–1688
Authors: Moore Peter N.
Published: April 2020

Historians on both sides of the Atlantic have failed to appreciate the significance of Stuarts Town, Scotland's short-lived colony in Port Royal, South Carolina. This article challenges the current view that Stuarts Town was primarily a business venture, focusing, instead, on the religious impulses that lay just beneath the surface of the Carolina Company. These concerns came to the fore as presbyterian persecution intensified in 1683 and the colony was reimagined as a safe haven for the true church, where the saving remnant of God's people could escape the terrible judgments befalling Scotland and where the gospel would be secure. Its purpose was collective, corporate, social and historical. On the ground in Carolina, however, colonisers behaved more like imperialists than religious refugees. Like Scotland, the Anglo-Spanish borderland was a violent and unstable place that bred fear of displacement and enslavement, but unlike Scotland it lacked a centralised power, giving the Scots an opening to make their bid for empire. They moved aggressively into this power vacuum, seeking in particular to capitalise on the perceived weakness of Spanish Florida to extend their reach into coastal Georgia, the south-eastern interior and as far west as New Mexico. Their actions created great anxiety in the region and, although the collapse of the Stuart regime finally put an end to their hopes, their short-lived colony transformed the borderlands, reorienting English, Spanish and Indian relations, sparking the coalescence of the Yamasee tribe and the Creek confederacy, and giving new life to the Indian slave trade that eventually shattered indigenous societies in the American south-east.
ADAPTIVE PLANNING OF THE PRODUCTION ORGANIZATION INDUSTRIAL ENTERPRISE
Authors: Babushkin Vitaly Mikhailovich , Sharafeev Ilgizar Shayheevich , Valiullin Rafik Zuferovich , Mingaleev Gaziz Fuatovich
Published: April 2020

The article presents an approach to planning the organization of production, in which the increase in productivity can be considered as a result of the implementation of predictive models of data Analytics and forecasting of events at each level of planning. Considered and justified mathematical model of optimization of resources of the enterprise, implemented in a three- tier system adaptive planning with the formation of indicators, optimization of production and allowing it to form the most effective scenarios of solving the problem maximize performance subject to qualitative changes in the organization of production related scientific and technical progress and forecasting trends. The formulated conceptual apparatus allows us to consider productivity as a function that reflects the impact on the production system of modern methods of planning and information technologies of production organization in time, which significantly expands the possibilities of theoretical research in the field of production organization. The three-level system of adaptive planning described in this article contributes to the development of the theory of optimizing the use of enterprise resources and the organization of efficient production. The application of the approach to adaptive planning of production organization presented in the article allows to expand the range of solutions to improve the productivity of enterprises by promptly responding to changes in its performance using modern technologies of lean and digital production.
STUDY OF THE INFLUENCE OF THE SPECTRAL COMPOSITION OF IR RADIATION ON THE SEDIMENTATION RATE OF COMPLEX COMPOUNDS
Authors: Rakhimov Rustam Khakimovich , Hasanov Ramil Zijaevich , Yermakov Vladimir Petrovich , Rakhimov Murod Rustamovich , Latipov Rustam Naimzhanovich , Atamanova Mamlakat Umarovna , Mamajanova Umida Bachodirovna
Published: April 2020

The aim of this work is to study the effect of pulsed IR radiation generated by functional ceramics and cavitation on the processes of sedimentation and coagulation of extremely fine fractions. It is shown that these methods are complementary for solving technological problems.
DEVELOPMENT OF MEMBRANE-ELECTRODE ASSEMBLIES PROTOTYPES BASED PLATINUM NANOCOMPOSITES FOR ENERGY SOURCES
Authors: Lebedeva Marina Vladimirovna , Antropov Alexey Petrovich , Ragutkin Alexander Victorovich , Yashtulov Nicolay Andreevich
Published: April 2020

Electrode materials with platinum nanoparticles on combined polymer-carbon matrix-carriers had been formed. Model tests of hydrogen-air fuel cells with varying loading and sizes of platinum nanoparticles had been carried out. It was found that the maximum value of specific power (64 mW/cm2) and current density (122-128 mA/cm2) are achieved when the catalyst load was 0.32 mg/cm2 and the nanoparticle size was 2-4 nm.
Investigation of a CubeSat in Orbit Anomaly through Verification on Ground
Authors: Stesina Fabrizio , Corpino Sabrina
Published: April 2020

Given the role of Cubesats in the new space economy, a statistically relevant number of CubeSats have flown, and considering the high percentage of failed missions, the investigation of in-orbit anomalies becomes of paramount importance. It is rare to find data about mission failures, probably because the partial or total absence of telemetry does not encourage any analysis. The lack of data from the spacecraft in orbit can be mitigated through ad-hoc verification campaigns on satellite models when in-orbit anomalies are experienced. This paper shows an effective testing activity conducted on models of the spacecraft to understand the root cause of a severe anomaly that occurred during mission operations. The tests are part of a comprehensive methodology for root causes analysis. The paper aims at sharing the experience built upon a practical case of interest. More importantly, this work has the ambition of fostering the research on key topics of reliability, mission operations and assembly, and integration and verification/test processes, which have shown to be critical. The activity presented in this paper demonstrates that investigating the anomalies can help recover the mission of interest but can also support building a heritage that is still missing for CubeSat missions today.
Wheat Straw Biochar and NPK Fertilization Efficiency in Sandy Soil Reclamation
Authors: Bednik Magdalena , Medyńska-Juraszek Agnieszka , Dudek Michał , Kloc Szymon , Kręt Agata , Łabaz Beata , Waroszewski Jarosław
Published: April 2020

Intensive land use including cultivation may result in soil degradation. Restoring natural conditions or utility values called reclamation usually requires prior improvement of soil properties by using fertilizers or soil amendments. Special attention is paid to biochar, as it can improve soil physical and chemical properties. Therefore, amendment is potentially useful for preparation of degraded soil for future vegetation. In pot experiments, we studied the effect of wheat straw biochar (5% v/w) (soil with biochar (S + BC)), two rates of mineral fertilizer (no fertilizer as a control set (SC) and single initial dose of fertilizer (S + NPK)), and combination of biochar and fertilizer (S + B C + NPK) on sandy soil properties and grass growth. Biochar significantly increased soil pH, total organic carbon content (TOC), and volumetric water content (VWC, +24% after one week of measurements). However, dry mass of grass shoots was lower in S + BC than in SC (−38%). It was also observed that, in fertilizer, applied sets high concentrations of salts caused crust formation, surface cracking, and overdrying. Considering the results, biochar may be useful for increasing sandy substrate fertility, providing proper conditions for revegetation during reclamation.
Antimicrobial Resistance in Farm Animals in Brazil: An Update Overview
Authors: Rabello Renata F. , Bonelli Raquel R. , Penna Bruno A. , Albuquerque Julia P. , Souza Rossiane M. , Cerqueira Aloysio M. F.
Published: April 2020

In animal husbandry, antimicrobial agents have been administered as supplements to increase production over the last 60 years. Large-scale animal production has increased the importance of antibiotic management because it may favor the evolution of antimicrobial resistance and select resistant strains. Brazil is a significant producer and exporter of animal-derived food. Although Brazil is still preparing a national surveillance plan, several changes in legislation and timely programs have been implemented. Thus, Brazilian data on antimicrobial resistance in bacteria associated with animals come from official programs and the scientific community. This review aims to update and discuss the available Brazilian data on this topic, emphasizing legal aspects, incidence, and genetics of the resistance reported by studies published since 2009, focusing on farm animals and derived foods with the most global public health impact. Studies are related to poultry, cattle, and pigs, and mainly concentrate on non-typhoid Salmonella, Escherichia coli, and Staphylococcus aureus. We also describe legal aspects of antimicrobial use in this context; and the current occurrence of genetic elements associated with resistance to beta-lactams, colistin, and fluoroquinolones, among other antimicrobial agents. Data here presented may be useful to provide a better understanding of the Brazilian status on antimicrobial resistance related to farm animals and animal-derived food products.
Analysis and Design of Rateless Two-way Relay Networks Based on a Multiply-and-Forward Scheme
Authors: Han Wei , Xu Shengkai , Huang Daqing , Xu Cheng
Published: April 2020

In this paper, we propose a rateless, three-stage, two-way multiply-and-forward (MF) relaying system over the Rayleigh flat fading channel, where two source nodes communicate with each other through a relay node and all nodes work on half-duplex and time-division mode. We thoroughly analyze the signals during all three stages in the proposed MF system and derive the closed-form symbol error rate (SER) expressions for an uncoded MF-two-way relay network (MF-TWRN). Furthermore, we provide the equivalent point-to-point fading channel model, which is employed to carry out the asymptotic performance analysis. We finally put forth an optimization model for the MF-TWRN with fountain codes. Simulation results show that our optimized degree distribution can provide outstanding performance for the MF-TWRN compared to those in the literature.
Passivity-Based Control Design for Magnetic Levitation System
Authors: Yang Wang , Meng Fanwei , Sun Man , Liu Kai
Published: April 2020

The passivity-based control (PBC) is a new direction of nonlinear control, but the method is basically a qualitative method. A quantifiable design method in combination with PBC is provided in this paper. To solve the partial differential equation (PDE) for PBC, the nonlinear system must first be transformed into a Hamiltonian model. The PDE for the Hamiltonian system is then quantifiably solved with an electromagnetic levitation example. The resulting control law is presented and discussed. The proposed method provides a practical design tool for nonlinear control.
Laser Welding on Cerclage Wire Joining in Femur Surgery
Authors: Thongchuea Nutchanat , Warinsiriruk Eakkachai , Wang Yin-Tien
Published: April 2020

Stainless steel wires with fiber-laser welding on lap joints are first proposed in this study to be used as cerclage wire joints for modern femur surgery, because of their potentially larger joint strength and less loosening failure than traditional joint devices. In this feasibility study, an experiment was set up to determine adequate parameters for the laser welding process in order to ensure that the wire joint of cerclage has good weld appearance, free oxidation, and suitable joint strength. A stainless steel wire 316L with a diameter of 1.6 mm, flare-welded on lap joint was used in the experiment as a specimen cerclage wire joint. Two major effective parameters were chosen for controlling a suitable fusion weld, including charge voltage and multiple frequencies of the laser irradiation. The adequate area of the laser parameters was determined from the experiment, including the ranges of charge voltage, multiple frequencies, and pulse width. The suitable welded lengths of specimens were also studied in the mechanical test to validate the joint strength. Suggested welded length has a better tensile strength than traditional cerclage joints. The paper concludes that the stainless wire joints with a fiber laser weld represents a promising alternative to traditional cerclage joint devices for modern femur surgery.
Improving Continuous Variable Quantum Secret Sharing with Weak Coherent States
Authors: Wang Yijun , Jia Bing , Mao Yun , Wu Xuelin , Guo Ying
Published: April 2020

Quantum secret sharing (QSS) can usually realize unconditional security with entanglement of quantum systems. While the usual security proof has been established in theoretics, how to defend against the tolerable channel loss in practices is still a challenge. The traditional ( t , n ) threshold schemes are equipped in situation where all participants have equal ability to handle the secret. Here we propose an improved ( t , n ) threshold continuous variable (CV) QSS scheme using weak coherent states transmitting in a chaining channel. In this scheme, one participant prepares for a Gaussian-modulated coherent state (GMCS) transmitted to other participants subsequently. The remaining participants insert independent GMCS prepared locally into the circulating optical modes. The dealer measures the phase and the amplitude quadratures by using double homodyne detectors, and distributes the secret to all participants respectively. Special t out of n participants could recover the original secret using the Lagrange interpolation and their encoded random numbers. Security analysis shows that it could satisfy the secret sharing constraint which requires the legal participants to recover message in a large group. This scheme is more robust against background noise due to the employment of double homodyne detection, which relies on standard apparatuses, such as amplitude and phase modulators, in favor of its potential practical implementations.
Potential Study of Solar Thermal Cooling in Sub-Mediterranean Climate
Authors: Jaradat Mustafa , Al-Addous Mohammad , Albatayneh Aiman , Dalala Zakariya , Barbana Nesrine
Published: April 2020

Air conditioning is becoming increasingly important in the energy supply of buildings worldwide. There has been a dramatic increase in energy requirements for cooling buildings in the Middle East and North Africa (MENA) region. This is before taking the effects of climate change into account, which will also entail a sharp increase in cooling requirements. This paper presents the potential of using a solar thermal absorption cooling system in Sub-Mediterranean Climate. Four sites in Jordan are now equipped with water-lithium bromide (H2O-LiBr) absorption chillers with a total nominal capacity of 530 kW. The focus of the paper was on the pilot system at the German Jordanian University (GJU) campus with a cooling capacity of 160 kW. The system was designed and integrated in order to support two existing conventional compression chillers with a nominal cooling capacity of 700 kW. The system was economically evaluated based on the observed cooling capacity results with a Coefficient of Performance (COP) equals 0.32, and compared with the values observed for a COP of 0.79 which is claimed by the manufacturer. Several techniques were implemented to evaluate the overall economic viability in-depth such as present worth value, internal rate of return, payback period, and levelized cost of electricity. The aforementioned economic studies showed that the absorption cooling system is deemed not feasible for the observed COP of 0.32 over a lifespan of 25 years. The net present value was equal to −137,684 JD and a payback period of 44 years which exceeds the expected lifespan of the project. Even for an optimal operation of COP = 0.79, the discounted payback period was equal to 23 years and the Levelized Cost of Electricity (LCOE) was equal to 0.65 JD/kWh. The survey shows that there are several weaknesses for applying solar thermal cooling in developing countries such as the high cost of these systems and, more significantly, the lack of experience for such systems.
BET Inhibitors Synergize with Carfilzomib to Induce Cell Death in Cancer Cells via Impairing Nrf1 Transcriptional Activity and Exacerbating the Unfolded Protein Response
Authors: Vangala Janakiram R. , Potluri Ajay , Radhakrishnan Senthil K.
Published: April 2020

Currently, proteasome inhibitors bortezomib, carfilzomib, and ixazomib are successfully used in clinics to treat multiple myeloma. However, these agents show limited efficacy against solid tumors. Identification of drugs that can potentiate the action of proteasome inhibitors could help expand the use of this therapeutic modality to solid tumors. Here, we found that bromodomain extra-terminal (BET) family protein inhibitors such as JQ1, I-BET762, and I-BET151 synergize with carfilzomib in multiple solid tumor cell lines. Mechanistically, BET inhibitors attenuated the ability of the transcription factor Nrf1 to induce proteasome genes in response to proteasome inhibition, thus, impeding the bounce-back response of proteasome activity, a critical pathway by which cells cope with proteotoxic stress. Moreover, we found that treatment with BET inhibitors or depletion of Nrf1 exacerbated the unfolded protein response (UPR), signaling that was initiated by proteasome inhibition. Taken together, our work provides a mechanistic explanation behind the synergy between proteasome and BET inhibitors in cancer cell lines and could prompt future preclinical and clinical studies aimed at further investigating this combination.
Exploring Alzheimer’s Disease Molecular Variability via Calculation of Personalized Transcriptional Signatures
Authors: Dagan Hila , Flashner-Abramson Efrat , Vasudevan Swetha , Jubran Maria R. , Cohen Ehud , Kravchenko-Balasha Nataly
Published: April 2020

Despite huge investments and major efforts to develop remedies for Alzheimer’s disease (AD) in the past decades, AD remains incurable. While evidence for molecular and phenotypic variability in AD have been accumulating, AD research still heavily relies on the search for AD-specific genetic/protein biomarkers that are expected to exhibit repetitive patterns throughout all patients. Thus, the classification of AD patients to different categories is expected to set the basis for the development of therapies that will be beneficial for subpopulations of patients. Here we explore the molecular heterogeneity among a large cohort of AD and non-demented brain samples, aiming to address the question whether AD-specific molecular biomarkers can progress our understanding of the disease and advance the development of anti-AD therapeutics. We studied 951 brain samples, obtained from up to 17 brain regions of 85 AD patients and 22 non-demented subjects. Utilizing an information-theoretic approach, we deciphered the brain sample-specific structures of altered transcriptional networks. Our in-depth analysis revealed that 7 subnetworks were repetitive in the 737 diseased and 214 non-demented brain samples. Each sample was characterized by a subset consisting of ~1–3 subnetworks out of 7, generating 52 distinct altered transcriptional signatures that characterized the 951 samples. We show that 30 different altered transcriptional signatures characterized solely AD samples and were not found in any of the non-demented samples. In contrast, the rest of the signatures characterized different subsets of sample types, demonstrating the high molecular variability and complexity of gene expression in AD. Importantly, different AD patients exhibiting similar expression levels of AD biomarkers harbored distinct altered transcriptional networks. Our results emphasize the need to expand the biomarker-based stratification to patient-specific transcriptional signature identification for improved AD diagnosis and for the development of subclass-specific future treatment.
Variations in Circulating Active MMP-9 Levels during Renal Replacement Therapy
Authors: Rodríguez-Sánchez Elena , Navarro-García José Alberto , Aceves-Ripoll Jennifer , Abarca-Zabalía Judith , Susmozas-Sánchez Andrea , Bada-Bosch Teresa , Hernández Eduardo , Mérida-Herrero Evangelina , Andrés Amado , Praga Manuel , Fernández-Ruiz Mario , Aguado José María , Segura Julián , Ruilope Luis Miguel , Ruiz-Hurtado Gema
Published: April 2020

Renal replacement therapy (RRT) is complicated by a chronic state of inflammation and a high mortality risk. However, different RRT modalities can have a selective impact on markers of inflammation and oxidative stress. We evaluated the levels of active matrix metalloproteinase (MMP)-9 in patients undergoing two types of dialysis (high-flux dialysis (HFD) and on-line hemodiafiltration (OL-HDF)) and in kidney transplantation (KT) recipients. Active MMP-9 was measured by zymography and ELISA before (pre-) and after (post-) one dialysis session, and at baseline and follow-up (7 and 14 days, and 1, 3, 6, and 12 months) after KT. Active MMP-9 decreased post-dialysis only in HFD patients, while the levels in OL-HDF patients were already lower before dialysis. Active MMP-9 increased at 7 and 14 days post-KT and was restored to baseline levels three months post-KT, coinciding with an improvement in renal function and plasma creatinine. Active MMP-9 correlated with pulse pressure as an indicator of arterial stiffness both in dialysis patients and KT recipients. In conclusion, active MMP-9 is better controlled in OL-HDF than in HFD and is restored to baseline levels along with stabilization of renal parameters after KT. Active MMP-9 might act as a biomarker of arterial stiffness in RRT.
A Novel Sweetpotato WRKY Transcription Factor, IbWRKY2, Positively Regulates Drought and Salt Tolerance in Transgenic Arabidopsis
Authors: Zhu Hong , Zhou Yuanyuan , Zhai Hong , He Shaozhen , Zhao Ning , Liu Qingchang
Published: April 2020

WRKYs play important roles in plant growth, defense regulation, and stress response. However, the mechanisms through which WRKYs are involved in drought and salt tolerance have been rarely characterized in sweetpotato [Ipomoea batatas (L.) Lam.]. In this study, we cloned a WRKY gene, IbWRKY2, from sweetpotato and its expression was induced with PEG6000, NaCl, and abscisic acid (ABA). The IbWRKY2 was localized in the nucleus. The full-length protein exhibited transactivation activity, and its active domain was located in the N-terminal region. IbWRKY2-overexpressing Arabidopsis showed enhanced drought and salt tolerance. After drought and salt treatments, the contents of ABA and proline as well as the activity of superoxide dismutase (SOD) were higher in transgenic plants, while the malondialdehyde (MDA) and H2O2 contents were lower. In addition, several genes related to the ABA signaling pathway, proline biosynthesis, and the reactive oxygen species (ROS)-scavenging system, were significantly up-regulated in transgenic lines. These results demonstrate that IbWRKY2 confers drought and salt tolerance in Arabidopsis. Furthermore, IbWRKY2 was able to interact with IbVQ4, and the expression of IbVQ4 was induced by drought and salt treatments. These results provide clues regarding the mechanism by which IbWRKY2 contributes to the regulation of abiotic stress tolerance.
Activation of α7 Nicotinic Acetylcholine Receptor Upregulates HLA-DR and Macrophage Receptors: Potential Role in Adaptive Immunity and in Preventing Immunosuppression
Authors: Siniavin Andrei , Streltsova Maria , Kudryavtsev Denis , Shelukhina Irina , Utkin Yuri , Tsetlin Victor
Published: April 2020

Immune response during sepsis is characterized by hyper-inflammation followed by immunosuppression. The crucial role of macrophages is well-known for both septic stages, since they are involved in immune homeostasis and inflammation, their dysfunction being implicated in immunosuppression. The cholinergic anti-inflammatory pathway mediated by macrophage α7 nicotinic acetylcholine receptor (nAChR) represents possible drug target. Although α7 nAChR activation on macrophages reduces the production of proinflammatory cytokines, the role of these receptors in immunological changes at the cellular level is not fully understood. Using α7 nAChR selective agonist PNU 282,987, we investigated the influence of α7 nAChR activation on the expression of cytokines and, for the first time, of the macrophage membrane markers: cluster of differentiation 14 (CD14), human leukocyte antigen-DR (HLA-DR), CD11b, and CD54. Application of PNU 282,987 to THP-1Mϕ (THP-1 derived macrophages) cells led to inward ion currents and Ca2+ increase in cytoplasm showing the presence of functionally active α7 nAChR. Production of cytokines tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-10 was estimated in classically activated macrophages (M1) and treatment with PNU 282,987 diminished IL-10 expression. α7 nAChR activation on THP-1Mϕ, THP-1M1, and monocyte-derived macrophages (MDMs) increased the expression of HLA-DR, CD54, and CD11b molecules, but decreased CD14 receptor expression, these effects being blocked by alpha (α)-bungarotoxin. Thus, PNU 282,987 enhances the macrophage-mediated immunity via α7 nAChR by regulating expression of their membrane receptors and of cytokines, both playing an important role in preventing immunosuppressive states.
A Novel Serine Protease Inhibitor PE-BBI Ameliorates Cockroach Extract-Mediated Airway Epithelial Barrier Dysfunction
Authors: Reihill James A. , Ouyang Xuan , Yang Zhixuan , Douglas Lisa E. J. , Zhou Mei , Chen Tianbao , Martin S. Lorraine
Published: April 2020

Epithelial barrier dysfunction, characteristic of allergic airway disease may be, at least in part, due to the action of allergen-associated protease activities. Cockroach allergy is a major global health issue, with cockroaches containing considerable serine trypsin-like protease (TLP) activity. The present study sought to evaluate two novel protease inhibitors (PE-BBI and pLR-HL), recently isolated from amphibian skin secretions, for their potential to neutralise cockroach TLP activity and to determine any protective effect on cockroach-induced airway epithelial barrier disruption. Inhibitor potencies against the cockroach-associated activities were determined using a fluorogenic peptide substrate-based activity assay. 16HBE14o- cells (16HBE; a bronchial epithelial cell line) were treated with cockroach extract (CRE) in the presence or absence of the compounds in order to assess cell viability (RealTime Glo luminescent assay) and epithelial barrier disruption (transepithelial resistance and paracellular dextran flux). PE-BBI potently and selectively inhibited CRE TLP activity (pIC50 -8), but not host (16HBE) cell surface activity, which conferred protection of 16HBE cells from CRE-induced cell damage and barrier disruption. Novel protease inhibitor strategies such as PE-BBI may be useful for the treatment of allergic airway disease caused by cockroach proteases.
A High-Throughput Single-Clone Phage Fluorescence Microwell Immunoassay and Laser-Driven Clonal Retrieval System
Authors: Chang Seohee , Kim Soohyun , Han Jerome , Ha Suji , Lee Hyunho , Song Seo Woo , Lee Daewon , Kwon Sunghoon , Chung Junho , Kim Junhoi
Published: April 2020

Phage display is one of the most frequently used platform technologies utilized to screen and select therapeutic antibodies, and has contributed to the development of more than 10 therapeutic antibodies used in the clinic. Despite advantages like efficiency and low cost, it has intrinsic technical limitations, such as the asymmetrical amplification of the library after each round of biopanning, which is regarded as a reason for it yielding a very limited number of antigen binders. In this study, we developed a high-throughput single-clonal screening system comprised of fluorescence immunoassays and a laser-driven clonal DNA retrieval system using microchip technology. Using this system, from a single-chain variable fragment (scFv) library displayed on phages with a complexity of 5.21 × 105 harboring random mutations at five amino acid residues, more than 70,000 clones—corresponding to ~14% of the library complexity—were screened, resulting in 78 antigen-reactive scFv sequences with mutations restricted to the randomized residues. Our results demonstrate that this system can significantly reduce the number of biopanning rounds, or even eliminate the need for this process for libraries with lower complexity, providing an opportunity to obtain more diverse clones from the library.
Uncovering Prognosis-Related Genes and Pathways by Multi-Omics Analysis in Lung Cancer
Authors: Asada Ken , Kobayashi Kazuma , Joutard Samuel , Tubaki Masashi , Takahashi Satoshi , Takasawa Ken , Komatsu Masaaki , Kaneko Syuzo , Sese Jun , Hamamoto Ryuji
Published: April 2020

Lung cancer is one of the leading causes of death worldwide. Therefore, understanding the factors linked to patient survival is essential. Recently, multi-omics analysis has emerged, allowing for patient groups to be classified according to prognosis and at a more individual level, to support the use of precision medicine. Here, we combined RNA expression and miRNA expression with clinical information, to conduct a multi-omics analysis, using publicly available datasets (the cancer genome atlas (TCGA) focusing on lung adenocarcinoma (LUAD)). We were able to successfully subclass patients according to survival. The classifiers we developed, using inferred labels obtained from patient subtypes showed that a support vector machine (SVM), gave the best classification results, with an accuracy of 0.82 with the test dataset. Using these subtypes, we ranked genes based on RNA expression levels. The top 25 genes were investigated, to elucidate the mechanisms that underlie patient prognosis. Bioinformatics analyses showed that the expression levels of six out of 25 genes (ERO1B, DPY19L1, NCAM1, RET, MARCH1, and SLC7A8) were associated with LUAD patient survival (p < 0.05), and pathway analyses indicated that major cancer signaling was altered in the subtypes.
(20S)G-Rh2 Inhibits NF-κB Regulated Epithelial-Mesenchymal Transition by Targeting Annexin A2
Authors: Wang Yu-Shi , Li He , Li Yang , Zhang Shiyin , Jin Ying-Hua
Published: April 2020

(1) Background: Epithelial-mesenchymal transition (EMT) is an essential step for cancer metastasis; targeting EMT is an important path for cancer treatment and drug development. NF-κB, an important transcription factor, has been shown to be responsible for cancer metastasis by enhancing the EMT process. Our previous studies showed that (20S)Ginsenoside Rh2 (G-Rh2) inhibits NF-κB activity by targeting Anxa2, but it is still not known whether this targeted inhibition of NF-κB can inhibit the EMT process. (2) Methods: In vivo (20S)G-Rh2-Anxa2 interaction was assessed by cellular thermal shift assay. Protein interaction was determined by immuno-precipitation analysis. NF-κB activity was determined by dual luciferase reporter assay. Gene expression was determined by RT-PCR and immuno-blot. EMT was evaluated by wound healing and Transwell assay and EMT regulating gene expression. (3) Results: Anxa2 interacted with the NF-κB p50 subunit, promoted NF-κB activation, then accelerated mesenchymal-like gene expression and enhanced cell motility; all these cellular processes were inhibited by (20S)G-Rh2. In contrast, these (20S)G-Rh2 effect were completely eliminated by overexpression of Anxa2-K301A, an (20S)G-Rh2-binding-deficient mutant of Anxa2. (4) Conclusion: (20S)G-Rh2 inhibited NF-κB activation and related EMT by targeting Anxa2 in MDA-MB-231 cells.
Development of Prostate Cancer Organoid Culture Models in Basic Medicine and Translational Research
Authors: Elbadawy Mohamed , Abugomaa Amira , Yamawaki Hideyuki , Usui Tatsuya , Sasaki Kazuaki
Published: April 2020

Prostate cancer (PC) is the most prevalent cancer in men and the second main cause of cancer-related death in Western society. The lack of proper PC models that recapitulate the molecular and genomic landscape of clinical disease has hampered progress toward translational research to understand the disease initiation, progression, and therapeutic responses in each patient. Although several models have been developed, they hardly emulated the complicated PC microenvironment. Precision medicine is an emerging approach predicting appropriate therapies for individual cancer patients by means of various analyses of individual genomic profiling and targeting specific cancer pathways. In PC, precision medicine also has the potential to impose changes in clinical practices. Here, we describe the various PC models with special focus on PC organoids and their values in basic medicine, personalized therapy, and translational researches in vitro and in vivo, which could help to achieve the full transformative power of cancer precision medicine.
Treatment of Combined Hepatocellular and Cholangiocarcinoma
Authors: Leoni Simona , Sansone Vito , De Lorenzo Stefania , Ielasi Luca , Tovoli Francesco , Renzulli Matteo , Golfieri Rita , Spinelli Daniele , Piscaglia Fabio
Published: April 2020

Combined hepatocellular and cholangiocarcinoma (HCC-CC) is a rare primary liver cancer. It is constituted by neoplastic cells of both hepatocellular and cholangiocellular derivation. Different histology types of HCC-CC have been reported, hinting at heterogeneous carcinogenic pathways leading to the development of this cancer. Due to its rarity and complexity, mixed HCC-CC is a scantly investigated condition with unmet needs and unsatisfactory outcomes. Surgery remains the preferred treatment in resectable patients. The risk of recurrence, however, is high, especially in comparison with other primary liver cancers such as hepatocellular carcinoma. In unresectable or recurring patients, the therapeutic options are challenging due to the dual nature of the neoplastic cells. Consequently, the odds of survival of patients with HCC-CC remains poor. We analysed the literature systematically about the treatment of mixed HCC-CC, reviewing the main therapeutic options and their outcomes and analysing the most interesting developments in this topic with a focus on new potential therapeutic avenues.
An Organotypic Microcosm for the Pancreatic Tumor Microenvironment
Authors: Lin Miranda , Gao Mei , Pandalai Prakash K. , Cavnar Michael J. , Kim Joseph
Published: April 2020

Pancreatic duct adenocarcinoma (PDAC) is projected to become the second leading cause of cancer-related deaths in the next few years. Unfortunately, the development of novel therapies for PDAC has been challenged by a uniquely complex tumor microenvironment. The development of in vitro cancer organoids in recent years has demonstrated potential to increase therapies for patients with PDAC. Organoids have been established from PDAC murine and human tissues and they are representative of the primary tumor. Further, organoids have been shown beneficial in studies of molecular mechanisms and drug sensitivity testing. This review will cover the use of organoids to study PDAC development, invasiveness, and therapeutic resistance in the context of the tumor microenvironment, which is characterized by a dense desmoplastic reaction, hindered immune activity, and pro-tumor metabolic signaling. We describe investigations utilizing organoids to characterize the tumor microenvironment and also describe their limitations. Overall, organoids have great potential to serve as a versatile model of drug response and may be used to increase available therapies and improve survival for patients with PDAC.
Personalized Medicine—Current and Emerging Predictive and Prognostic Biomarkers in Colorectal Cancer
Authors: Koulis Christine , Yap Raymond , Engel Rebekah , Jardé Thierry , Wilkins Simon , Solon Gemma , Shapiro Jeremy D. , Abud Helen , McMurrick Paul
Published: April 2020

Colorectal cancer (CRC) is the third most common cancer diagnosed worldwide and is heterogeneous both morphologically and molecularly. In an era of personalized medicine, the greatest challenge is to predict individual response to therapy and distinguish patients likely to be cured with surgical resection of tumors and systemic therapy from those resistant or non-responsive to treatment. Patients would avoid futile treatments, including clinical trial regimes and ultimately this would prevent under- and over-treatment and reduce unnecessary adverse side effects. In this review, the potential of specific biomarkers will be explored to address two key questions—1) Can the prognosis of patients that will fare well or poorly be determined beyond currently recognized prognostic indicators? and 2) Can an individual patient’s response to therapy be predicted and those who will most likely benefit from treatment/s be identified? Identifying and validating key prognostic and predictive biomarkers and an understanding of the underlying mechanisms of drug resistance and toxicity in CRC are important steps in order to personalize treatment. This review addresses recent data on biological prognostic and predictive biomarkers in CRC. In addition, patient cohorts most likely to benefit from currently available systemic treatments and/or targeted therapies are discussed in this review.
Ensartinib (X-396) Effectively Modulates Pharmacokinetic Resistance Mediated by ABCB1 and ABCG2 Drug Efflux Transporters and CYP3A4 Biotransformation Enzyme
Authors: Vagiannis Dimitrios , Novotna Eva , Skarka Adam , Kammerer Sarah , Küpper Jan-Heiner , Chen Si , Guo Lei , Staud Frantisek , Hofman Jakub
Published: April 2020

Ensartinib (X-396) is a promising tyrosine kinase inhibitor currently undergoing advanced clinical evaluation for the treatment of non-small cell lung cancer. In this work, we investigate possible interactions of this promising drug candidate with ATP-binding cassette (ABC) drug efflux transporters and cytochrome P450 biotransformation enzymes (CYPs), which play major roles in multidrug resistance (MDR) and pharmacokinetic drug-drug interactions (DDIs). Accumulation studies showed that ensartinib is a potent inhibitor of ABCB1 and ABCG2 transporters. Additionally, incubation experiments with recombinant CYPs showed that ensartinib significantly inhibits CYP3A4 and CYP2C9. Subsequent molecular docking studies confirmed these findings. Drug combination experiments demonstrated that ensartinib synergistically potentiates the antiproliferative effects of daunorubicin, mitoxantrone, and docetaxel in ABCB1, ABCG2, and CYP3A4-overexpressing cellular models, respectively. Advantageously, ensartinib’s antitumor efficiency was not compromised by the presence of MDR-associated ABC transporters, although it acted as a substrate of ABCB1 in Madin-Darby Canine Kidney II (MDCKII) monolayer transport assays. Finally, we demonstrated that ensartinib had no significant effect on the mRNA-level expression of examined transporters and enzymes in physiological and lung tumor cellular models. In conclusion, ensartinib may perpetrate clinically relevant pharmacokinetic DDIs and modulate ABCB1-, ABCG2-, and CYP3A4-mediated MDR. The in vitro findings presented here will provide a valuable foundation for future in vivo investigations.
NK and NKT Cell-Mediated Immune Surveillance against Hematological Malignancies
Authors: Shimizu Kanako , Iyoda Tomonori , Yamasaki Satoru , Kadowaki Norimitsu , Tojo Arinobu , Fujii Shin-ichiro
Published: April 2020

Recent cancer treatment modalities have been intensively focused on immunotherapy. The success of chimeric antigen receptor T cell therapy for treatment of refractory B cell acute lymphoblastic leukemia has pushed forward research on hematological malignancies. Among the effector types of innate lymphocytes, natural killer (NK) cells show great importance in immune surveillance against infectious and tumor diseases. Particularly, the role of NK cells has been argued in either elimination of target tumor cells or escape of tumor cells from immune surveillance. Therefore, an NK cell activation approach has been explored. Recent findings demonstrate that invariant natural killer T (iNKT) cells capable of producing IFN-γ when optimally activated can promptly trigger NK cells. Here, we review the role of NKT and/or NK cells and their interaction in anti-tumor responses by highlighting how innate immune cells recognize tumors, exert effector functions, and amplify adaptive immune responses. In addition, we discuss these innate lymphocytes in hematological disorders, particularly multiple myeloma and acute myeloid leukemia. The immune balance at different stages of both diseases is explored in light of disease progression. Various types of innate immunity-mediated therapeutic approaches, recent advances in clinical immunotherapies, and iNKT-mediated cancer immunotherapy as next-generation immunotherapy are then discussed.
Therapy-Induced Senescence: An “Old” Friend Becomes the Enemy
Authors: Saleh Tareq , Bloukh Sarah , Carpenter Valerie J. , Alwohoush Enas , Bakeer Jomana , Darwish Sarah , Azab Belal , Gewirtz David A.
Published: April 2020

For the past two decades, cellular senescence has been recognized as a central component of the tumor cell response to chemotherapy and radiation. Traditionally, this form of senescence, termed Therapy-Induced Senescence (TIS), was linked to extensive nuclear damage precipitated by classical genotoxic chemotherapy. However, a number of other forms of therapy have also been shown to induce senescence in tumor cells independently of direct genomic damage. This review attempts to provide a comprehensive summary of both conventional and targeted anticancer therapeutics that have been shown to induce senescence in vitro and in vivo. Still, the utility of promoting senescence as a therapeutic endpoint remains under debate. Since senescence represents a durable form of growth arrest, it might be argued that senescence is a desirable outcome of cancer therapy. However, accumulating evidence suggesting that cells have the capacity to escape from TIS would support an alternative conclusion, that senescence provides an avenue whereby tumor cells can evade the potentially lethal action of anticancer drugs, allowing the cells to enter a temporary state of dormancy that eventually facilitates disease recurrence, often in a more aggressive state. Furthermore, TIS is now strongly connected to tumor cell remodeling, potentially to tumor dormancy, acquiring more ominous malignant phenotypes and accounts for several untoward adverse effects of cancer therapy. Here, we argue that senescence represents a barrier to effective anticancer treatment, and discuss the emerging efforts to identify and exploit agents with senolytic properties as a strategy for elimination of the persistent residual surviving tumor cell population, with the goal of mitigating the tumor-promoting influence of the senescent cells and to thereby reduce the likelihood of cancer relapse.
Landscape and Future Perspectives of Immunotherapy in Neuroendocrine Neoplasia
Authors: Maggio Ilaria , Manuzzi Lisa , Lamberti Giuseppe , Ricci Angela Dalia , Tober Nastassja , Campana Davide
Published: April 2020

Background: Neuroendocrine neoplasms are rare entities consisting of a heterogeneous group of tumors that can originate from neuroendocrine cells present in the whole body. Their different behavior, metastatic potential, and prognosis are highly variable, depending on site of origin, grade of differentiation, and proliferative index. The aim of our work is to summarize the current knowledge of immunotherapy in different neuroendocrine neoplasms and its implication in clinical practice. Results: Several studies evaluated the efficacy and safety of immunotherapy in neuroendocrine neoplasms, in any setting of treatment, alone or in combination. Studies led to approval in neuroendocrine neoplasia of the lung, in combination with chemotherapy as first-line treatment or as a single-agent in a third-line setting, and Merkel cell carcinoma as a single agent. Results in other settings have been disappointing so far. Conclusions: Immunotherapy seems a valid treatment option for high grade, poorly differentiated neoplasms. Future trials should explore the combination of immunotherapy with other agents, such as anti-angiogenic or other immunotherapy agents, in order to evaluate potential efficacy in low and intermediate grades, well differentiated tumors.
Preoperative Peak Oxygen Consumption: A Predictor of Survival in Resected Lung Cancer
Authors: Lindenmann Joerg , Fink-Neuboeck Nicole , Fediuk Melanie , Maier Alfred , Kovacs Gabor , Balic Marija , Smolle Josef , Smolle-Juettner Freyja Maria
Published: April 2020

The peak oxygen consumption (VO2 peak) serves as a prognostic factor in cardio-respiratory diseases and plays an important role in cancer patients. The long-term prognostic relevance of VO2 peak in lung cancer patients has not been investigated extensively. The aim of this study was to evaluate the impact of the preoperative VO2 peak on the postoperative long-term survival in patients with operated lung cancer. Retrospective analysis of 342 patients with curatively resected non-small-cell lung cancer using a multivariate Cox proportional hazard model. Results: Preoperative VO2 peak ranged from 10.2 to 51.8 mL/kg/min (mean: 18.3 ± 4.6), VO2 peak % of predicted ranged from 32 to 172% (mean: 65.2 ± 18.0%). Overall 10-year survival was 23%. A Log-rank test comparing predicted VO2 peak ≥ 60% with predicted VO2 peak < 60% showed overall survival of 30% and 17%, respectively (p < 0.001) and non-tumour-related survival of 71% and 51% (p = 0.001) at 10 years. In multivariable Cox analysis, overall 10-year survival correlated with a high predicted VO2 peak% (p = 0.001) and low N-stage corresponding to N0 and N1 (p < 0.001). Non-tumour-related death correlated with low VO2 peak% of predicted (p = 0.001), and age (p < 0.001). Low preoperative VO2 peak was associated with both decreased postoperative overall survival and decreased non-tumour-related survival during the 10-year follow-up.