bluuebunny/crossref_metadata_2025 huggingface dataset:
The Inhibition of Wnt Restrain KRASG12V-Driven Metastasis in Non-Small-Cell Lung Cancer
Authors: Hung Pei-Shan , Huang Ming-Hung , Kuo Yuan-Yeh , Yang James Chih-Hsin
Published: April 2020

The KRAS mutations have been an obstacle to identify therapeutic targets in cancer treatment. In this work, we clarified the distinct metastasis pattern of non-small-cell lung carcinoma (NSCLC) induced by KRASG12V/KRASG12D mutations and inhibited the KRASG12V mediated metastasis by Wnt inhibitor. First, we found that KRASG12V induced more aggressive phenotype in vitro and in vivo experiments. The Gene Set Enrichment Analysis (GSEA) results of H838 KRASG12V cells showed a significant negative correlation with RhoA-related signaling. Following this clue, we observed KRASG12D induced higher activation of RhoA and suppressed activation of Wnt/β-catenin in H838KRASG12D cells. The restored activation of Wnt/β-catenin in H838KRASG12D cells could be detected when expression with a dominant-negative mutant of RhoA or treatment with RhoA inhibitor. Furthermore, the Wnt inhibitor abolished the KRASG12V-induced migration. We elucidated the importance of the axis of RhoA/Wnt in regulatory NSCLC metastasis driven by KRAS mutations. Our data indicate that KRASG12V driven NSCLC metastasis is Wnt-dependent and the mechanisms of NSCLC metastasis induced by KRASG12V/KRASG12D is distinct.
Effectiveness and Costs Associated to Adding Cetuximab or Bevacizumab to Chemotherapy as Initial Treatment in Metastatic Colorectal Cancer: Results from the Observational FABIO Project
Authors: Franchi Matteo , Garau Donatella , Kirchmayer Ursula , Di Martino Mirko , Romero Marilena , De Carlo Ilenia , Scondotto Salvatore , Corrao Giovanni
Published: April 2020

Evidence available on the effectiveness and costs of biological therapies for the initial treatment of metastatic colorectal cancer (mCRC) is scarce and contrasting. We conducted a population-based cohort investigation for assessing overall survival and costs associated with their use in a real-world setting. Healthcare utilization databases were used to select patients newly diagnosed with mCRC between 2010 and 2016. Those initially treated with biological therapy (bevacizumab or cetuximab) added to chemotherapy were propensity-score-matched to those treated with standard chemotherapy alone, and were followed up to June 30th, 2018. Kaplan–Meier survival estimates, restricted mean survival time (RMST) and cumulative costs were compared between the two treatment arms. The study cohort included 1896 mCRC patients treated with biological therapy matched to 5678 patients treated with chemotherapy alone. Median overall survival was 21.8 and 20.2 months, respectively. After 84 months of follow-up, RMSTs were 30.9 and 31.9 months (p = 0.193), indicating no differences between the average survival time between treatment arms. Patients treated with biological therapy were associated with higher costs. Cumulative per capita costs were €59,663 and €44,399, respectively. In our study, first-line biological therapy did not improve long-term overall survival and was associated with higher costs as compared to standard chemotherapy.
Safety of Aflibercept in Metastatic Colorectal Cancer: A Literature Review and Expert Perspective on Clinical and Real-World Data
Authors: Muro Kei , Salinardi Taylor , Singh Arvind Rup , Macarulla Teresa
Published: April 2020

Background: Metastatic colorectal cancer (mCRC) represents a substantial health burden globally and an increasing challenge in Asian countries. Treatment options include chemotherapy plus a vascular endothelial growth factor (VEGF) inhibitor (such as bevacizumab, aflibercept or ramucirumab), or anti-epidermal growth factor receptor (EGFR) therapies. Aflibercept, a recombinant fusion protein, has been approved for treatment of mCRC in combination with FOLFIRI for patients whose disease progresses during or after treatment with an oxaliplatin-containing regimen, based on its efficacy and tolerability profile in clinical trials. This report aims to provide an overview of both clinical and real-world evidence and experience on the use of aflibercept in routine clinical practice, with a focus on European, American and Asian populations. Methods: A literature search was conducted in PubMed (on 28th February 2019) using the search terms ("aflibercept") and ("Colorectal"OR"CRC") to identify publications containing information on aflibercept-containing regimens. Results: The adverse events (AE) profile was similar between geographical locations. Across trials, real-world and retrospective studies, grade ≥ 3 hypertension and proteinuria were amongst the most frequently reported AEs. Conclusions: The safety profile of aflibercept is generally manageable and comparable across various geographic locations.
Lactotransferrin Downregulation Drives the Metastatic Progression in Clear Cell Renal Cell Carcinoma
Authors: Chiu I-Jen , Hsu Yung-Ho , Chang Jeng-Shou , Yang Jou-Chun , Chiu Hui-Wen , Lin Yuan-Feng
Published: April 2020

Clear cell renal cell carcinoma (ccRCC) is the main type of RCC, which is the most common type of malignant kidney tumor in adults. A subpopulation (>30%) of ccRCC patients develop metastasis; however, the molecular mechanism remains largely unknown. Here, we found that LTF, the gene encoding lactotransferrin, is dramatically downregulated in primary tumors compared to normal tissues derived from ccRCC patients deposited in The Cancer Genome Atlas (TCGA) database and is a favorable prognostic marker. Moreover, LTF downregulation appears to be more dominant in metastatic ccRCC. LTF overexpression suppresses migration ability in A498 ccRCC cells with high metastatic potential, whereas LTF knockdown fosters cellular migration in poorly metastatic ccRCC cells. Gene set enrichment analysis demonstrated that LTF expression inversely correlates with the progression of epithelial-mesenchymal transition (EMT) in ccRCC, which was further confirmed by RT-PCR experiments. Therapeutically, the administration of recombinant LTF protein significantly suppresses the cell migration ability and lung metastatic potential of ACHN cells, as well as LTF-silenced A498 cells. The gene knockdown of lipoprotein receptor-related protein 1 (LRP1) robustly blocked recombinant LTF protein-induced inhibition of cellular migration and gene expression of EMT markers in ACHN cells. LTF downregulation and LRP1 upregulation combined predicted a poor overall survival rate in ccRCC patients compared to that with either factor alone. Our findings uncover a new mechanism by which LTF may interact with LRP1 to inhibit metastatic progression in ccRCC and also reveal the therapeutic value of recombinant LTF protein in treating metastatic ccRCC.
Morphologic and Genomic Heterogeneity in the Evolution and Progression of Breast Cancer
Authors: Kutasovic Jamie R. , McCart Reed Amy E. , Sokolova Anna , Lakhani Sunil R. , Simpson Peter T.
Published: April 2020

Breast cancer is a remarkably complex and diverse disease. Subtyping based on morphology, genomics, biomarkers and/or clinical parameters seeks to stratify optimal approaches for management, but it is clear that every breast cancer is fundamentally unique. Intra-tumour heterogeneity adds further complexity and impacts a patient’s response to neoadjuvant or adjuvant therapy. Here, we review some established and more recent evidence related to the complex nature of breast cancer evolution. We describe morphologic and genomic diversity as it arises spontaneously during the early stages of tumour evolution, and also in the context of treatment where the changing subclonal architecture of a tumour is driven by the inherent adaptability of tumour cells to evolve and resist the selective pressures of therapy.
Enhanced Photocatalytic Hydrogen Generation by Optimized Plasmonic Hot Electron Injection in Structure-Adjustable Au-ZnO Hybrids
Authors: Chen Youlong , Ma Liang , Ding Sijing
Published: April 2020

Plasmonic Au-ZnO hybrids with adjustable structures (including Au-decorated ZnO and core–shell Au@ZnO with dense and porous ZnO shells) and the optimized hot electron-driven photocatalytic activity were successfully prepared. It was found that the Au@ZnO core–shell hybrids with porous morphology had the highest plasmon-enhanced photocatalytic hydrogen generation activity under visible light irradiation. The wavelength-dependent photocatalytic tests verified that Au@ZnO with porous ZnO shells had the highest apparent quantum efficiency upon resonance excitation. The ultrafast transient absorption measurements revealed that Au@ZnO with porous ZnO shells had the fastest plasmon-induced hot electron injection, which was thought to be the reason for the improved photocatalytic activity. This work might provide a promising route to designing photocatalytic and photoelectric materials.
Shaking Rate during Production Affects the Activity of Escherichia coli Surface-Displayed Candida antarctica Lipase A
Authors: Chung Chen-Fu , Lin Shih-Che , Juang Tzong-Yuan , Liu Yung-Chuan
Published: April 2020

In this study, a surface-display system was applied for the expression of lipase A in an E. coli expression system. Since the target protein was exposed on the cell membrane, the shaking rate during culturing might have increased the oxygen mass transfer rate and the shear stress, both of which would be detrimental to the surface-displayed protein. The shaking rate did indeed have an effect on the properties of the surface-displayed lipase A from Candida antarctica (sdCALA). When cultivated at a shaking rate of less than 50 rpm, the specific activity of sdCALA was low, which was due to the limited amount of dissolved oxygen. When the shaking rate was greater than 100 rpm, the specific activity decreased as a result of shear stress. When cultivating CALA and sdCALA at various temperatures and values of pH, both proteins displayed the same activity profile, with the optimum conditions being 60 °C and pH 6. A kinetic study revealed that the sdCALA cultivated at 100 rpm gave a higher value of νm (0.074 μmol/mL/min) and a lower value of Km (0.360 μmol/mL) relative to those obtained at 200 rpm and relative to those of the free CALA. sdCALA retained over 80% of its activity after treatment at 70 °C for 30 min, but its activity decreased rapidly when the temperature was above 80 °C. The specific activity of sdCALA decreased in the presence of acetonitrile and acetone relative to that of the control (50% ethanol), regardless of the solvent concentration. The highest activity (0.67 U/mL) was obtained when the ethanol concentration was 30%.
Combined Effect of Photocatalyst, Superplasticizer, and Glass Fiber on the Photocatalytic Activity and Technical Parameters of Gypsum
Authors: Zając Kamila , Czyżewski Adam , Kaszyńska Maria , Janus Magdalena
Published: April 2020

In this study, gypsum plasters enriched with the modified photocatalyst TiO2/N and additive components were prepared and analyzed in detail. The aim of this work was to recognize the combined impact of a photocatalyst, polycarboxylic superplasticizer (SP), and glass fiber (F) on the properties of the composed building materials. The mutual compatibility was verified in relation to self-cleaning and air-purifying activity as well as to the technical parameters defined in standards for gypsum materials. The measurements revealed that photocatalytic gypsum material can have high mechanical strength and limited shrinkage as a result of superplasticizer contribution in form of a water-reducing and well-dispersive agent. Normal consistency was achieved by the addition of 0.01 wt % of SP to photocatalytic gypsum mortar or by the addition of 0.2 wt % of SP with a 12% reduction of water. This study also explains why glass fiber fulfills the role of inner reinforcement only if a superplasticizer is simultaneously added to the gypsum matrix. It is possible, by the combined effect of TiO2/N, a polycarboxylic superplasticizer, and glass fiber, to achieve NOx degradation at a high level. Moreover, the significantly improved self-cleaning properties of the complex gypsum plasters surfaces from dye pollutants in comparison to a gypsum plaster with solely a TiO2/N photocatalyst indicate the synergistic effect of the three considered additives.
± 180° Discontinuous PWM for Single-Phase PWM Converter of High-Speed Railway Propulsion System
Authors: Song Min-Sup , Cho In-Ho , Lee Jae-Bum
Published: April 2020

As high-capacity alternating current/direct current (ac/dc) power conversion systems, single-phase pulse-width modulation (PWM) converters used in high-speed railway propulsion systems adopt high-voltage Insulated-Gate Bipolar Transistors (IGBTs) as switching elements. Due to their high breakdown voltage characteristics, the switching dynamics are inferior to those of low-voltage IGBTs and switching losses are more dominant than conduction losses despite operating at relatively low switching frequencies of hundreds to several kHz. To solve this problem, this paper proposes ± 180° discontinuous PWM (DPWM) suitable for a single-phase circuit. With the simple addition of offset voltages, the proposed DPWM method can be implemented easily and switching losses can be reduced by half by clamping the switching legs of the H-bridge converter to the positive or negative dc rail during every half cycle. In addition, temperature deviation between the power stacks can be minimized by using selective application of clamping modes. The validity and effectiveness of the proposed DPWM are verified through simulations and experiments of a prototype converter.
Numerical Analysis of Aerodynamic Characteristics of Exhaust Passage with Consideration of Wet Steam Effect in a Supercritical Steam Turbine
Authors: Xu Qing , Lin Aqiang , Cai Yuhang , Ahmad Naseem , Duan Yu , Liu Chen
Published: April 2020

To investigate the aerodynamic performance of exhaust passage under multi-phase flow, an actual case is conducted in the low-pressure double exhaust passages of 600 MW steam turbine. Then, the flow field is compared and analyzed with and without the built-in extraction pipelines based on the Eulerian–Eulerian homogenous medium multiphase method. Results show that the upstream swirling flow and downstream mixed swirling flow are the main causes to induce the entropy-increase in the exhaust passage. Moreover, the flow loss and static-pressure recovery ability in the exhaust hood are greater than those in the condenser neck. Compared with the flow field without the steam extraction pipelines, the entropy-increase increases, the static pressure recovery coefficient decreases, and the spontaneous condensation rates of wet steam decrease in the downstream area of the pipelines. With the increase of steam turbine loads, an increment in entropy-increase in the exhaust passage is 0.98 J/(kg·K) lower than that without steam extraction pipelines. Moreover, the incrementing range of uniformity coefficient is increased from 14.5% to 40.9% at the condenser neck outlet. It can be concluded that the built-in exhaustion pipeline can improve the aerodynamic performance of exhaust passage and better reflect the real state of the flow field. These research results can serve as a reference for turbine passage design.
Transcriptome Sequencing and Differential Expression Analysis Reveal Molecular Mechanisms for Starch Accumulation in Chestnut
Authors: Li Shengxing , Liang Haiying , Tao Liang , Xiong Liquan , Liang Wenhui , Shi Zhuogong , Zhao Zhiheng
Published: April 2020

Chestnuts are popular edible nuts that are rich in starch. In order to enhance the transcriptomic resources and further understand starch and sucrose metabolism in maturing chestnuts, a comparative transcriptomic study of Chinese chestnut kernels was conducted at three ripening stages (70, 82, and 94 DAF). At 82 and 94 days after flowering (DAF), starch continued to accumulate, and the amylopectin/amylose ratio increased. Transcriptomic profiling of kernels at 70 (stage I), 82 (stage II), and 94 DAF (stage III) indicated that soluble starch synthase and α-1,4-glucan branching enzyme genes are actively expressed at 82 and 94 DAF. The starch degradation enzymes amylase, phosphoglucan phosphatase DSP4, and maltose exporter did not show differential gene expression, while glycogen phosphorylase-encoding unigenes were significantly down-regulated at 94 DAF. In addition to starch and sucrose metabolism, RNA transport, RNA degradation, pyrimidine metabolism, purine metabolism, plant hormone signal transduction, plant–pathogen interactions, and glycerophospholipid metabolism were found to be significantly enriched in all comparisons included in the study. As Chinese chestnut matured, the unique enriched pathways switched from ribosomal biogenesis and RNA polymerase of eukaryotes to endocytosis and spliceosomes. These genomic resources and findings are valuable for further understanding starch and sucrose metabolism in the Chinese chestnut.
The Effects of Oxidation on the Antithrombotic Properties of Tea Lipids against PAF, Thrombin, Collagen, and ADP
Authors: Tsoupras Alexandros , Lordan Ronan , Harrington Jack , Pienaar Rebecca , Devaney Karen , Heaney Stephanie , Koidis Anastasios , Zabetakis Ioannis
Published: April 2020

Tea provides health benefits, while oxidation is part of tea processing. The effect of oxidation on the antithrombotic properties of tea lipid extracts was evaluated for the first time. Total lipids (TL) extracted from fresh tea leaves and commercial tea powder, before and after 30–60 min of oxidation, were further fractionated into neutral lipids (NL) and polar lipids (PL). The antithrombotic bioactivities of tea TL, PL, and NL were assessed in human platelets against the inflammatory mediator platelet-activating factor. PL were further assessed against thrombin, collagen, and adenosine diphosphate, while their fatty acid composition was evaluated by GC-MS. PL exhibited the strongest antithrombotic effects against all platelet agonists and were rich in omega-3 polyunsaturated (ω3 PUFA) and monounsaturated (MUFA) fatty acids. A decline was observed in the antithrombotic activities, against all platelet agonists tested, for PL after 60 min of oxidation, and on their MUFA content, while their overall ω3 PUFA content and ω6/ω3 ratio remained unaffected. A synergistic effect between tea phenolic compounds and PL protects them against oxidation, which seems to be the rational for retaining the antithrombotic biofunctionalities of PL at a considerable favorable cardioprotective level, even after 60 min of tea oxidation. More studies are required to elucidate the mechanisms of the favorable synergism in tea PL extracts.
Determination and Comparison of Physical Meat Quality Parameters of Percidae and Salmonidae in Aquaculture
Authors: Komolka Katrin , Bochert Ralf , Franz George P. , Kaya Yagmur , Pfuhl Ralf , Grunow Bianka
Published: April 2020

Although aquaculture has been the fastest growing food sector for decades, there are no standardized parameters for most of the fish species regarding physical meat quality. Therefore, this study provides for the first time an overview of the physical meat characteristics of the most important fish species of the German Baltic Sea coast. Traditional farmed salmonids (rainbow trout (Oncorhynchus mykiss) and maraena whitefish (Coregonus maraena) as well as two percids (European perch, Perca fluviatilis and pikeperch, Sander lucioperca) were utilized for this comparison. The results demonstrate that the meat of the salmonids is very analogous. However, the post mortem degradation process starts faster in trout meat. In contrast, the meat quality characteristics of the percids are relatively different. The meat of pikeperch has comparatively low shear strength with a high water-holding capacity resulting in high meat tenderness. The opposite situation is present in European perch. The results indicate that it is not possible to establish the overall quality characteristics for fish or production form, as there is a high range of variability. Consequently, it is particularly important that meat quality characteristics are developed for important aquaculture species for further improvement through changes in husbandry conditions when necessary.
Parametric Analysis on Local Mechanisms of Masonry Churches in Teramo (Italy)
Authors: Vaiano Generoso , Formisano Antonio , Fabbrocino Francesco
Published: April 2020

Local collapse mechanisms related to the out-of-plane response of walls are commonly observed in existing masonry buildings subjected to earthquakes. In such structures, the lack of proper connections among orthogonal walls and between walls and floors does not allow a global box-type behaviour of the building to develop, which would be governed by the in-plane response of walls. In this paper, parametric linear kinematic analyses on the main local mechanisms of masonry churches were performed with the aim to evaluate the corresponding horizontal load multipliers. This study was conducted on 12 masonry churches, located in Teramo (Italy) and affected by the 2016 Central Italy earthquake, whose main out-of-plane collapse mechanisms, namely facade overturning, vertical bending, corner overturning and roof gable wall overturning, have been analysed. For each mechanism, parametric analysis was carried out on varying heights and thicknesses of walls. Firstly, the acceleration values activating the considered mechanisms were calculated in order to conduct checks prescribed by the current Italian standard. Subsequently, on the basis of the obtained results, simple analytical procedures to determine load collapse multiplier for each mechanism were drawn. Finally, ranges of suitable values of both the thickness and height of walls were found in order to always satisfy seismic checks.
Entrepreneurial, Economic, and Social Well-Being Outcomes from an RCT of a Youth Entrepreneurship Education Intervention among Native American Adolescents
Authors: Tingey Lauren , Larzelere Francene , Goklish Novalene , Rosenstock Summer , Jennings Mayo-Wilson Larissa , Pablo Elliott , Goklish Warren , Grass Ryan , Sprengeler Feather , Parker Sean , Ingalls Allison , Craig Mariddie , Barlow Allison
Published: April 2020

Background: Entrepreneurship education has demonstrated positive impacts in low-resource contexts. However, there is limited evidence of such programs evaluated among Native American (NA) youth in a rural reservation. Methods: A 2:1 randomized controlled trial evaluated the impact of the Arrowhead Business Group (ABG) entrepreneurship education program on entrepreneurship knowledge, economic empowerment, and social well-being among 394 NA youth. An intent to treat analysis using mixed effects regression models examined within and between study group differences from baseline to 24 months. An interaction term measured change in the intervention relative to change in the control. ABG participants were purposively sampled to conduct focus groups and in-depth interviews. Results: Significant intervention vs. control group improvements were sustained at 12 months for entrepreneurship knowledge and economic confidence/security. Significant within-group improvements were sustained for ABG participants at 24 months for connectedness to parents, school, and awareness of connectedness. Qualitative data endorses positive impacts on social well-being among ABG participants. Conclusion: Observed effects on entrepreneurship knowledge, economic empowerment, and connectedness, supplemented by the experiences and changes as described by the youth themselves, demonstrates how a strength-based youth entrepreneurship intervention focused on developing assets and resources may be an innovative approach to dually address health and economic disparities endured in Native American communities.
Self-Selected Pacing During a World Record Attempt in 40 Ironman-Distance Triathlons in 40 Days
Authors: Sousa Caio Victor , Pereira Rinaldo Wellerson , Rosemann Thomas , Nikolaidis Pantelis T. , Knechtle Beat
Published: April 2020

The present case study analyzed performance, pacing, and potential predictors in a self-paced world record attempt of a professional triathlete to finish 40 Ironman-distance triathlons within 40 days. Split times (i.e., swimming, cycling, running) and overall times, body weight, daily highest temperature, wind speed, energy expenditure, mean heart rate, and sleeping time were recorded. Non-linear regressions were applied to investigate changes in split and overall times across days. Multivariate regression analyses were performed to test which variables showed the greatest influence on the dependent variables cycling, running and overall time. The athlete completed the 40×Ironman distances in a total time of 444:22 h:min. He spent 50:26 h:min in swimming, 245:37 h:min in cycling, 137:17 h:min in running and 11:02 h:min in transition times. Swimming and cycling times became slower across days, whereas running times got faster until the 20th day and, thereafter, became slower until the 40th day. Overall times got slower until the 15th day, became faster to 31st, and started then to get slower until the end. Wind speed, previous day’s race time and average heart race during cycling were significant independent variables influencing cycling time. Body weight and average heart rate during running were significant independent variables influencing running performance. Cycling performance, running performance, and body weight were significant independent variables influencing overall time. In summary, running time was influenced by body weight, cycling by wind speed, and overall time by both running and cycling performances.
Knockout of the OsNAC006 Transcription Factor Causes Drought and Heat Sensitivity in Rice
Authors: Wang Bo , Zhong Zhaohui , Wang Xia , Han Xiangyan , Yu Deshui , Wang Chunguo , Song Wenqin , Zheng Xuelian , Chen Chengbin , Zhang Yong
Published: April 2020

Rice (Oryza sativa) responds to various abiotic stresses during growth. Plant-specific NAM, ATAF1/2, and CUC2 (NAC) transcription factors (TFs) play an important role in controlling numerous vital growth and developmental processes. To date, 170 NAC TFs have been reported in rice, but their roles remain largely unknown. Herein, we discovered that the TF OsNAC006 is constitutively expressed in rice, and regulated by H2O2, cold, heat, abscisic acid (ABA), indole-3-acetic acid (IAA), gibberellin (GA), NaCl, and polyethylene glycol (PEG) 6000 treatments. Furthermore, knockout of OsNAC006 using the CRISPR-Cas9 system resulted in drought and heat sensitivity. RNA sequencing (RNA-seq) transcriptome analysis revealed that OsNAC006 regulates the expression of genes mainly involved in response to stimuli, oxidoreductase activity, cofactor binding, and membrane-related pathways. Our findings elucidate the important role of OsNAC006 in drought responses, and provide valuable information for genetic manipulation to enhance stress tolerance in future plant breeding programs.
Simple Selection Procedure to Distinguish between Static and Flexible Loops
Authors: Mitusińska Karolina , Skalski Tomasz , Góra Artur
Published: April 2020

Loops are the most variable and unorganized elements of the secondary structure of proteins. Their ability to shift their shape can play a role in the binding of small ligands, enzymatic catalysis, or protein–protein interactions. Due to the loop flexibility, the positions of their residues in solved structures show the largest B-factors, or in a worst-case scenario can be unknown. Based on the loops’ movements’ timeline, they can be divided into slow (static) and fast (flexible). Although most of the loops that are missing in experimental structures belong to the flexible loops group, the computational tools for loop reconstruction use a set of static loop conformations to predict the missing part of the structure and evaluate the model. We believe that these two loop types can adopt different conformations and that using scoring functions appropriate for static loops is not sufficient for flexible loops. We showed that common model evaluation methods, are insufficient in the case of flexible solvent-exposed loops. Instead, we recommend using the potential energy to evaluate such loop models. We provide a novel model selection method based on a set of geometrical parameters to distinguish between flexible and static loops without the use of molecular dynamics simulations. We have also pointed out the importance of water network and interactions with the solvent for the flexible loop modeling.
DNA Strand Break Properties of Protoporphyrin IX by X-ray Irradiation against Melanoma
Authors: Hasegawa Takema , Takahashi Junko , Nagasawa Shinsuke , Doi Motomichi , Moriyama Akihiro , Iwahashi Hitoshi
Published: April 2020

Recent reports have suggested that 5-aminolevulinic acid (5-ALA), which is a precursor to protoporphyrin IX (PpIX), leads to selective accumulation of PpIX in tumor cells and acts as a radiation sensitizer in vitro and in vivo in mouse models of melanoma, glioma, and colon cancer. In this study, we investigated the effect of PpIX under X-ray irradiation through ROS generation and DNA damage. ROS generation by the interaction between PpIX and X-ray was evaluated by two kinds of probes, 3′-(p-aminophenyl) fluorescein (APF) for hydroxyl radical (•OH) detection and dihydroethidium (DHE) for superoxide (O2•-). •OH showed an increase, regardless of the dissolved oxygen. Meanwhile, the increase in O2•- was proportional to the dissolved oxygen. Strand breaks (SBs) of DNA molecule were evaluated by gel electrophoresis, and the enhancement of SBs was observed by PpIX treatment. We also studied the effect of PpIX for DNA damage in cells by X-ray irradiation using a B16 melanoma culture. X-ray irradiation induced γH2AX, DNA double-strand breaks (DSBs) in the context of chromatin, and affected cell survival. Since PpIX can enhance ROS generation even in a hypoxic state and induce DNA damage, combined radiotherapy treatment with 5-ALA is expected to improve therapeutic efficacy for radioresistant tumors.
Evaluation of Antibacterial and Cytotoxicity Properties of Silver Nanowires and Their Composites with Carbon Nanotubes for Biomedical Applications
Authors: De Mori Arianna , Jones Richard S. , Cretella Matteo , Cerri Guido , Draheim Roger R. , Barbu Eugen , Tozzi Gianluca , Roldo Marta
Published: April 2020

In this work, we prepared silver nanowires (AgNWs) via the polyol method in the presence or absence of single wall carbon nanotubes (CNTs) and tested their physicochemical, antibacterial and cytotoxic properties. Results showed that the introduction of CNTs lead to the formation of AgNWs at lower temperature, but the final product characteristics of AgNWs and AgNWs-CNT were not significantly different. AgNWs exhibited antibacterial properties against all the studied bacterial species via the formation of oxygen reactive species (ROS) and membrane damage. Furthermore, AgNWs exhibited a dose-dependent and time-dependent toxicity at concentrations ≥ 10 µg/mL. Fibroblasts appeared to be more resistant than human colorectal adenocarcinoma (Caco-2) and osteoblasts to the toxicity of AgNWs. The cytotoxicity of AgNWs was found to be related to the formation of ROS, but not to membrane damage. Overall, these results suggest that AgNWs are potential antibacterial agents against E. coli, S. aureus, MRSA and S. saprophyticus, but their dosage needs to be adjusted according to the route of administration.
Radiolabeled Human Monoclonal Antibody 067-213 has the Potential for Noninvasive Quantification of CD73 Expression
Authors: Sudo Hitomi , Tsuji Atsushi B. , Sugyo Aya , Kurosawa Gene , Kurosawa Yoshikazu , Alexander David , Tsuda Hiroyuki , Saga Tsuneo , Higashi Tatsuya
Published: April 2020

Background: CD73 is an ectonucleotidase regulating extracellular adenosine concentration and plays an important role in adenosine-mediated immunosuppressive pathways. The efficacy of CD73-targeted therapy depends on the expression levels of CD73; therefore, monitoring CD73 status in cancer patients would provide helpful information for selection of patients who would benefit from CD73-targeted therapy. Here, we evaluated the ability of 111In-labeled antibody 067-213, which has high affinity for human CD73, to act as a noninvasive imaging probe. Methods: Cell binding and competitive inhibition assays for 111In-labeled 067-213 were conducted using MIAPaCa-2 (high CD73 expression) and A431 (low CD73 expression) cells. For in vivo assessments, biodistribution and SPECT/CT studies were conducted in MIAPaCa-2 and A431 tumor-bearing mice. To estimate the absorbed dose in humans, biodistribution and SPECT/CT studies were conducted in healthy rats. Results: 111In-labeled 067-213 bound to MIAPaCa-2 and A431 cells in a CD73-dependent manner and the affinity loss after 111In-labeling was limited. Biodistribution and SPECT/CT studies with 111In-labeled 067-213 in mice showed high uptake in MIAPaCa-2 tumors and lower uptake in A431 tumors. In rats, the probe did not show high uptake in normal organs, including endogenously CD73-expressing organs. The estimated absorbed doses in humans were reasonably low. Conclusions: 111In-labeled 067-213 showed CD73-expression-dependent tumor uptake and low uptake in normal organs and tissues. Radiolabeled 067-213 holds promise as an imaging probe for noninvasive evaluation of CD73 expression levels in patients. Our data encourage further clinical studies to clarify a role for CD73 monitoring in patients receiving CD73-targeted immune therapy.
Therapeutic Use of Extracellular Vesicles for Acute and Chronic Lung Disease
Authors: Worthington Erin N. , Hagood James S.
Published: April 2020

Multipotent mesenchymal stem cells (MSCs) possess regenerative properties and have been shown to improve outcomes and survival in acute and chronic lung diseases, but there have been some safety concerns raised related to MSC-based therapy. Subsequent studies have demonstrated that many of the regenerative effects of MSCs can be attributed to the MSC-derived secretome, which contains soluble factors and extracellular vesicles (EVs). MSC-derived extracellular vesicles (MSC-derived EVs) replicate many of the beneficial effects of MSCs and contain a variety of bioactive factors that are transferred to recipient cells, mediating downstream signaling. MSC-derived EV therapy holds promise as a safe and effective treatment for pulmonary disease, but there remain many scientific and clinical questions that will need to be addressed before EVs are widely applied as a therapy. To date, the use of MSC-derived EVs as a treatment for lung disease has been conducted primarily in in vitro or pre-clinical animal models. In this review, we will discuss the current published research investigating the use of EVs as a potential therapeutic for acute lung injury/acute respiratory distress syndrome (ALI/ARDS), bronchopulmonary dysplasia (BPD), idiopathic pulmonary fibrosis (IPF), pulmonary arterial hypertension (PAH), asthma, and silicosis.
Anti-Inflammatory and Anti-Migratory Activities of Isoquinoline-1-Carboxamide Derivatives in LPS-Treated BV2 Microglial Cells via Inhibition of MAPKs/NF-κB Pathway
Authors: Do Ha Thi Thu , Bui Bich Phuong , Sim Seongrak , Jung Jae-Kyung , Lee Heesoon , Cho Jungsook
Published: April 2020

Eleven novel isoquinoline-1-carboxamides (HSR1101~1111) were synthesized and evaluated for their effects on lipopolysaccharide (LPS)-induced production of pro-inflammatory mediators and cell migration in BV2 microglial cells. Three compounds (HSR1101~1103) exhibited the most potent suppression of LPS-induced pro-inflammatory mediators, including interleukin (IL)-6, tumor necrosis factor-alpha, and nitric oxide (NO), without significant cytotoxicity. Among them, only N-(2-hydroxyphenyl) isoquinoline-1-carboxamide (HSR1101) was found to reverse LPS-suppressed anti-inflammatory cytokine IL-10, so it was selected for further characterization. HSR1101 attenuated LPS-induced expression of inducible NO synthase and cyclooxygenase-2. Particularly, HSR1101 abated LPS-induced nuclear translocation of NF-κB through inhibition of IκB phosphorylation. Furthermore, HSR1101 inhibited LPS-induced cell migration and phosphorylation of mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase 1/2, c-Jun N-terminal kinase, and p38 MAPK. The specific MAPK inhibitors, U0126, SP600125, and SB203580, suppressed LPS-stimulated pro-inflammatory mediators, cell migration, and NF-κB nuclear translocation, indicating that MAPKs may be the upstream kinase of NF-κB signaling. Collectively, these results demonstrate that HSR1101 is a potent and promising compound suppressing LPS-induced inflammation and cell migration in BV2 microglial cells, and that inhibition of the MAPKs/NF-κB pathway mediates its anti-inflammatory and anti-migratory effects. Based on our findings, HSR1101 may have beneficial impacts on various neurodegenerative disorders associated with neuroinflammation and microglial activation.
Effects of an Aquaporin 4 Inhibitor, TGN-020, on Murine Diabetic Retina
Authors: Oosuka Shou , Kida Teruyo , Oku Hidehiro , Horie Taeko , Morishita Seita , Fukumoto Masanori , Sato Takaki , Ikeda Tsunehiko
Published: April 2020

Purpose: To investigate the effect of a selective aquaporin 4 (AQP4) inhibitor, 2-(nicotinamide)-1,3,4-thiadiazole (TGN-020), on the expression of vascular endothelial growth factor (VEGF) and reactive oxygen species (ROS) production, as well as on the retinal edema in diabetic retina. Methods: Intravitreal injections of bevacizumab, TGN-020, or phosphate-buffered saline (PBS) were performed on streptozotocin-induced diabetic rats. Retinal sections were immunostained for anti-glial fibrillary acidic protein (GFAP), anti-AQP4, and anti-VEGF. Protein levels of VEGF from collected retinas were determined by Western blot analysis. In addition, retinal vascular leakage of Evans Blue was observed in the flat-mounted retina from the diabetic rats in the presence or absence of TGN-020. Volumetric changes of rat retinal Müller cells (TR-MUL5; transgenic rat Müller cells) and intracellular levels of ROS were determined using flow cytometry analysis of ethidium fluorescence in the presence or absence of TGN-020 or bevacizumab under physiological and high glucose conditions. Results: In the diabetic retina, the immunoreactivity and protein levels of VEGF were suppressed by TGN-020. AQP4 immunoreactivity was higher than in the control retinas and the expressions of AQP4 were co-localized with GFAP. Similarly to VEGF, AQP4 and GFAP were also suppressed by TGN-020. In the Evans Blue assay, TGN-020 decreased leakage in the diabetic retinas. In the cultured Müller cells, the increase in cell volumes and intracellular ROS production under high glucose condition were suppressed by exposure to TGN-020 as much as by exposure to bevacizumab. Conclusion: TGN-020 may have an inhibitory effect on diabetic retinal edema.
Vitamin D Suppresses Ovarian Cancer Growth and Invasion by Targeting Long Non-Coding RNA CCAT2
Authors: Wang Liye , Zhou Shuang , Guo Bin
Published: April 2020

Ovarian cancer is the most deadly gynecologic cancer among women worldwide. Poor response to current treatment makes it necessary to discover new diagnostic biomarkers to detect the cancer early and develop new and effective prevention strategies. Calcitriol, the active metabolite of vitamin D, protects against multiple cancers through unelucidated mechanisms. The oncogenic long non-coding RNA (lncRNA) CCAT2 (colon cancer associated transcript 2) is overexpressed in ovarian cancer. Here, we foundd that calcitriol inhibited CCAT2 expression in ovarian cancer cell lines. Treatment with calcitriol inhibited ovarian cancer cell proliferation, migration, and invasion. As a result of CCAT2 inhibition, calcitriol decreased the binding of transcription factor TCF7L2 (TCF4) to the MYC promoter, resulting in the repression of c-Myc protein expression. Our results suggest a novel anti-cancer mechanism of vitamin D by targeting CCAT2 in ovarian cancer. The findings may help develop vitamin D as a practical and inexpensive nutraceutical for ovarian cancer prevention.
Differential Targeting of Gr-MDSCs, T Cells and Prostate Cancer Cells by Dactolisib and Dasatinib
Authors: Liu Guoqiang , Jin Zhijian , Lu Xin
Published: April 2020

Granulocytic myeloid-derived suppressor cells (Gr-MDSCs) promote immune evasion and resistance to immunotherapeutics in a variety of malignancies. Our previous study showed that dual PI3K/mTOR inhibitor Dactolisib impaired the viability and immunosuppressive function of Gr-MDSCs, and significantly synergized with immune checkpoint blockade (ICB) antibodies targeting PD1 and CTLA4 to eradicate metastatic castration-resistant prostate cancer (CRPC) in a preclinical transgenic mouse model. On the contrary, tyrosine kinase inhibitor Dasatinib diminished tumor-infiltrating T lymphocytes and showed no synergic activity with ICB. The understanding of the distinct effects of Dactolisib and Dasatinib on Gr-MDSCs, T cells and prostate neoplastic cells is inadequate, limiting the clinical translation of the combination immunotherapy. To address this question, we applied Reverse Phase Protein Array (RPPA) to profile 297 proteins and protein phosphorylation sites of Gr-MDSCs, T cells and prostate cancer cells isolated from the CRPC model. We found cell type-specific protein expression patterns and highly selective targets by the two drugs, including preferential inhibition of phospho-4E-BP1 in Gr-MDSCs by Dactolisib and preferential suppression of phospho-Src and phospho-p38 MAPK in T cells. Furthermore, transcriptomic profiling of Gr-MDSCs treated with the two inhibitors revealed downregulation of mitochondrial respiration pathways by Dactolisib but not Dasatinib. Overall, these results provide important mechanistic insight into the efficacious combination of Dactolisib and ICB as well as the detrimental effect of Dasatinib on anti-tumor immunity.
Orchid Biochemistry
Authors: Chen Jen-Tsung
Published: April 2020

Orchids belong to Orchidaceae which is one of the largest families in flowering plants [...]
Longitudinal Cognitive Decline in a Novel Rodent Model of Cerebral Amyloid Angiopathy Type-1
Authors: Popescu Dominique L. , Van Nostrand William E. , Robinson John K.
Published: April 2020

Cerebral amyloid angiopathy (CAA) is a small vessel disease characterized by β-amyloid (Aβ) accumulation in and around the cerebral blood vessels and capillaries and is highly comorbid with Alzheimer’s disease (AD). Familial forms of CAA result from mutations within the Aβ domain of the amyloid β precursor protein (AβPP). Numerous transgenic mouse models have been generated around expression of human AβPP mutants and used to study cerebral amyloid pathologies. While behavioral deficits have been observed in many AβPP transgenic mouse lines, relative to rats, mice are limited in behavioral expression within specific cognitive domains. Recently, we generated a novel rat model, rTg-DI, which expresses Dutch/Iowa familial CAA Aβ in brain, develops progressive and robust accumulation of cerebral microvascular fibrillar Aβ beginning at 3 months, and mimics many pathological features of the human disease. The novel rTg-DI model provides a unique opportunity to evaluate the severity and forms of cognitive deficits that develop over the emergence and progression of CAA pathology. Here, we present an in-depth, longitudinal study aimed to complete a comprehensive assessment detailing phenotypic disease expression through extensive and sophisticated operant testing. Cohorts of rTg-DI and wild-type (WT) rats underwent operant testing from 6 to 12 months of age. Non-operant behavior was assessed prior to operant training at 4 months and after completion of training at 12 months. By 6 months, rTg-DI animals demonstrated speed–accuracy tradeoffs that later manifested across multiple operant tasks. rTg-DI animals also demonstrated delayed reaction times beginning at 7 months. Although non-operant assessments at 4 and 12 months indicated comparable mobility and balance, rTg-DI showed evidence of slowed environmental interaction. Overall, this suggests a form of sensorimotor slowing is the likely core functional impairment in rTg-DI rats and reflects similar deficits observed in human CAA.
Compensation of Mutation in Arabidopsis glutathione transferase (AtGSTU) Genes under Control or Salt Stress Conditions
Authors: Horváth Edit , Bela Krisztina , Gallé Ágnes , Riyazuddin Riyazuddin , Csomor Gábor , Csenki Dorottya , Csiszár Jolán
Published: April 2020

Glutathione transferases (GSTs) play a crucial role in detoxification processes due to the fact of their glutathione (GSH) conjugating activity, and through glutathione peroxidase or dehydroascorbate reductase (DHAR) activities, they influence the redox state of GSH and ascorbate (AsA). The plant-specific tau (GSTU) group is the largest class of Arabidopsis GSTs, and their members are involved in responses to different abiotic stresses. We investigated the effect of salt stress on two-week-old Arabidopsis thaliana wild-type (Col-0), Atgstu19 and Atgstu24 mutant plants after applying 150 mM NaCl for two days. The Atgstu19 seedlings had lower GST activity and vitality both under control conditions and after salt stress than the wild-type, but the level of total ROS was similar to the Col-0 plants. The GST activity of the knockout Atgstu24 mutant was even higher under control conditions compared to the Col-0 plants, while the ROS level and its vitality did not differ significantly from the wild-type. Analysis of the AtGSTU expression pattern revealed that the mutation in a single AtGSTU gene was accompanied by the up- and downregulation of several other AtGSTUs. Moreover, elevated AsA and GSH levels, an altered GSH redox potential and increased DHAR and glutathione reductase activities could help to compensate for the mutation of AtGSTU genes. The observed changes in the mutants suggest that the investigated isoenzymes influence the redox homeostasis under control conditions and after NaCl treatment in Arabidopsis seedlings. These data indicate for the first time the more general role of a temporary shift of redox status as part of GST mechanisms and regulation.
BMAL1 Suppresses Proliferation, Migration, and Invasion of U87MG Cells by Downregulating Cyclin B1, Phospho-AKT, and Metalloproteinase-9
Authors: Gwon Do hyeong , Lee Woo-Yong , Shin Nara , Kim Song I , Jeong Kuhee , Lee Won-hyung , Kim Dong Woon , Hong Jinpyo , Lee Sun Yeul
Published: April 2020

Several studies have shown that brain and muscle aryl hydrocarbon receptor nuclear translocator-like 1 (BMAL1), an important molecule for maintaining circadian rhythms, inhibits the growth and metastasis of tumor cells in several types of cancer, including lung, colon, and breast cancer. However, its role in glioblastoma has not yet been established. Here, we addressed the function of BMAL1 in U87MG glioblastoma cells with two approaches—loss and gain of function. In the loss of function experiments, cell proliferation in U87MG cells transfected with small interfering RNA (siRNA) targeting BMAL1 was increased by approximately 24% (small interfering (si)-NC 0.91 ± 0.00 vs. si-BMAL1 1.129 ± 0.08) via upregulation of cyclin B1. In addition, cell migration and invasion of BMAL1 siRNA-treated glioblastoma cells were elevated by approximately 20% (si-NC 51.00 ± 1.53 vs. si-BMAL161.33 ± 0.88) and 209% (si-NC 21.28 ± 1.37 vs. si-BMAL1 44.47 ± 3.48), respectively, through the accumulation of phosphorylated-AKT (p-AKT) and matrix metalloproteinase (MMP)-9. Gain of function experiments revealed that adenovirus-mediated ectopic expression of BMAL1 in U87MG cells resulted in a 19% (Adenovirus (Ad)-vector 0.94± 0.03 vs. Ad-BMAL1 0.76 ± 0.03) decrease in cell proliferation compared with the control via downregulation of cyclin B1 and increased early and late apoptosis due to changes in the levels of BCL2-associated X protein (BAX), B-cell lymphoma 2 (BCL-2), and cleaved caspase-3. Likewise, cell migration and invasion were attenuated by approximately 24% (Ad-vector 55.00 ± 0.00 vs. Ad-BMAL1 41.83 ± 2.90) and 49% (Ad-vector 70.01 ± 1.24 vs. Ad-BMAL1 35.55 ± 1.78), respectively, in BMAL1-overexpressing U87MG cells following downregulation of p-AKT and MMP-9. Taken together, our results suggest that BMAL1 acts as an anti-cancer gene by altering the proliferation, migration, and invasion of glioblastoma cells. Therefore, the BMAL1 gene could be a potential therapeutic target in the treatment of glioblastoma.