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At least 1,099 records · Page 61

Identification of methyl violet 2B as a novel blocker of focal adhesion kinase signaling pathway in cancer cells

Highlights: •FAK signaling cascade in cancer cells is profoundly inhibited by methyl violet 2B. •Methyl violet 2B identified by virtual screening is a novel allosteric FAK inhibitor. •Methyl violet 2B possesses extremely high kinase selectivity. •Methyl violet 2B suppresses strongly the proliferation of cancer cells. •Methyl violet 2B inhibits focal adhesion, invasion and migration of cancer cells. -- Abstract: The focal adhesion kinase (FAK) signaling cascade in cancer cells was profoundly inhibited by methyl violet 2B identified with the structure-based virtual screening. Methyl violet 2B was shown to be a non-competitive inhibitor of full-length FAK enzyme vs. ATP. It turned out that methyl violet 2B possesses extremely high kinase selectivity in biochemical kinase profiling using a large panel of kinases. Anti-proliferative activity measurement against several different cancer cells and Western blot analysis showed that this substance is capable of suppressing significantly the proliferation of cancer cells and is able to strongly block FAK/AKT/MAPK signaling pathways in a dose dependent manner at low nanomolar concentration. Especially, phosphorylation of Tyr925-FAK that is required for full activation of FAK was nearly completely suppressed even with 1 nM of methyl violet 2B in A375P cancer cells. To the best of our knowledge, it has never been reported that methyl violet possesses anti-cancer effects. Moreover, methyl violet 2B significantly inhibited FER kinase phosphorylation that activates FAK in cell. In addition, methyl violet 2B was found to induce cell apoptosis and to exhibit strong inhibitory effects on the focal adhesion, invasion, and migration of A375P cancer cells at low nanomolar concentrations. Taken together, these results show that methyl violet 2B is a novel, potent and selective blocker of FAK signaling cascade, which displays strong anti-proliferative activities against a variety of human cancer cells and suppresses adhesion/migration/invasion of tumor cells.

60 APPLIED LIFE SCIENCES↗

GPR30 decreases cardiac chymase/angiotensin II by inhibiting local mast cell number

Chronic activation of the novel estrogen receptor GPR30 by its agonist G1 mitigates the adverse effects of estrogen (E2) loss on cardiac structure and function. Using the ovariectomized (OVX) mRen2.Lewis rat, an E2-sensitive model of diastolic dysfunction, we found that E2 status is inversely correlated with local cardiac angiotensin II (Ang II) levels, likely via Ang I/chymase-mediated production. Since chymase is released from cardiac mast cells during stress (e.g., volume/pressure overload, inflammation), we hypothesized that GPR30-related cardioprotection after E2 loss might occur through its opposing actions on cardiac mast cell proliferation and chymase production. Using real-time quantitative PCR, immunohistochemistry, and immunoblot analysis, we found mast cell number, chymase expression, and cardiac Ang II levels were significantly increased in the hearts of OVX-compared to ovary-intact mRen2.Lewis rats and the GPR30 agonist G1 (50 mg/kg/day, s.c.) administered for 2 weeks limited the adverse effects of estrogen loss. In vitro studies revealed that GPR30 receptors are expressed in the RBL-2H3 mast cell line and G1 inhibits serum-induced cell proliferation in a dose-dependent manner, as determined by cell counting, BrdU incorporation assay, and Ki-67 staining. Using specific antagonists to estrogen receptors, blockage of GPR30, but not ERα or ERβ, attenuated the inhibitory effects of estrogen on BrdU incorporation in RBL-2H3 cells. Further study of the mechanism underlying the effect on cell proliferation showed that G1 inhibits cyclin-dependent kinase 1 (CDK1) mRNA and protein expression in RBL-2H3 cells in a dose-dependent manner. - Highlights: • GPR30 activation limits mast cell number in hearts from OVX mRen2.Lewis rats. • GPR30 activation decreases cardiac chymase/angiotensin II after estrogen loss. • GPR30 activation inhibits RBL-2H3 mast cell proliferation and CDK1 expression.

60 APPLIED LIFE SCIENCES↗

2-Deoxy-d-glucose increases GFAT1 phosphorylation resulting in endoplasmic reticulum-related apoptosis via disruption of protein N-glycosylation in pancreatic cancer cells

Highlights: • 2DG causes ER stress-induced cell death by disrupting protein N-glycosylation. • This is caused by inactivation of GFAT1 in pancreatic cancer. • In addition, combination treatment with 2DG and Met enhanced disruption of protein N-glycosylation. The glycolytic inhibitor 2-deoxy-d-glucose (2DG) causes energy starvation, affecting cell viability in a wide range of cancer cell lines. To determine the action of 2DG in pancreatic cancer, we performed proteomic analysis of pancreatic cancer cell line after 2DG treatment. Eighty proteins showed differential expression and among these, proteins involved in phosphohexose metabolism were upregulated. Up-regulation of glutamine: fructose 6-phosphate aminotransferase 1 (GFAT1), which belongs to the hexosamine biosynthesis pathway (HBP) that produces uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) to maintain glycoprotein, was validated by evaluation of mRNA and protein levels. Therefore, we assessed the amounts of total N-glycoproteins. Unexpectedly, we found a reduction of total N-glycoproteins and phosphorylation of GFAT1 by AMP-activated protein kinase (AMPK). These data may shed light on HBP dysfunction. Furthermore, we found endoplasmic reticulum (ER) stress accompanied by increased expression of ER stress markers, such as glucose response protein 78 (GRP78) and C/EBP-homologous protein (CHOP), in 2DG-treated cells. Moreover, the additive activation of AMPK by metformin (Met) synergistically enhanced the reduction of protein N-glycosylation and cell growth inhibition in the presence of 2DG. These results suggest that 2DG reduces N-glycosylation of proteins following the increase of phosphorylation of GFAT1 and results in the inhibition of cell growth mediated by ER stress in pancreatic cancer cells.

60 APPLIED LIFE SCIENCES↗

Role of p-GaN layer thickness in the degradation of InGaN-GaN MQW solar cells under 405 nm laser excitation

GaN-based solar cells with InGaN multiple quantum wells (MQWs) are promising devices for application in space environment, concentrator solar systems, wireless power transmission and multi-junction solar cells. It is therefore important to understand their degradation kinetics when submitted to high-temperature and high-intensity stress. We submitted three samples of GaN-InGaN MQW solar cells with p-AlGaN electron-blocking-layer with different thickness of the p-GaN layer to constant power stress at 310 W/cm 2 , 175 °C for several hundred hours. The main degradation modes are a reduction of open-circuit voltage, short-circuit current, external quantum efficiency, power conversion efficiency and electroluminescence. In particular, we observed that a thinner p-GaN layer results in a stronger degradation observed on the cell operating parameters. The analysis of the dark I-V characteristics showed an increase in low-forward bias current and the analysis of electroluminescence showed a decrease in the electroluminescene emitted by the (forward biased) cell, as a consequence of stress. This work highlights that the cause of degradation is possibly related to a diffusion mechanism, which results in an increase of defect density in the active region. Finally, the impurities involved in the diffusion processes possibly originate from the p-side of the devices, therefore a thicker p-GaN layer reduces the amount of defects reaching the active region.

14 SOLAR ENERGY↗

Development and Application of TK6-derived Cells Expressing Human Cytochrome P450s for Genotoxicity Testing

Abstract Metabolism plays a key role in chemical genotoxicity; however, most mammalian cells used for in vitro genotoxicity testing lack effective metabolizing enzymes. We recently developed a battery of TK6-derived cell lines that individually overexpress 1 of 8 cytochrome P450s (CYP1A1, 1A2, 1B1, 2A6, 2B6, 2C9, 2C19, and 3A4) using a lentiviral expression system. The increased expression and metabolic function of each individual CYP in each established cell line were confirmed using real-time PCR, Western blotting, and mass spectrometry analysis; the parental TK6 cells and empty vector (EV) transduced cells had negligible CYP levels. Subsequently, we evaluated these cell lines using 2 prototypical polyaromatic hydrocarbon mutagens, 7,12-dimethylbenz[a]anthracene (DMBA) and benzo[a]pyrene (B[a]P), that require metabolic activation to exert their genotoxicity. DMBA-induced cytotoxicity, phosphorylation of histone H2A.X, and micronucleus formation were significantly increased in TK6 cells with CYP1A1, 1B1, 2B6, and 2C19 expression as compared with EV controls. B[a]P significantly increased cytotoxicity, DNA damage, and chromosomal damage in TK6 cells overexpressing CYP1A1 and 1B1 when compared with EV controls. B[a]P also induced micronucleus formation in TK6 cells expressing CYP1A2. These results suggest that our CYP-expressing TK6 cell system can be used to detect the genotoxicity of compounds requiring metabolic transformation.

Li, Xilin↗

Optimizing solar-cell grid geometry

Trade-off analysis and mathematical expressions calculate optimum grid geometry in terms of various cell parameters. Determination of the grid geometry provides proper balance between grid resistance and cell output to optimize the energy conversion process.

Crossley, A. P.↗

Review of physics underlying recent improvements in silicon solar-cell performance

This paper provides a unifying view of the physics of silicon solar cells, and uses it as a basis for explaining how recent improvements in the performance of these cells have been achieved. The unification is facilitated by a region-by-region analysis of the solar cell, which is also used to compare several recently proposed cell structures.

Lindholm, F. A.↗

Device research task (processing and high-efficiency solar cells)

This task has been expanded since the last 25th Project Integration Meeting (PIM) to include process research in addition to device research. The objective of this task is to assist the Flat-plate Solar Array (FSA) Project in meeting its near- and long-term goals by identifying and implementing research in the areas of device physics, device structures, measurement techniques, material-device interactions, and cell processing. The research efforts of this task are described and reflect the deversity of device research being conducted. All of the contracts being reported are either completed or near completion and culminate the device research efforts of the FSA Project. Optimazation methods and silicon solar cell numerical models, carrier transport and recombination parameters in heavily doped silicon, development and analysis of silicon solar cells of near 20% efficiency, and SiN sub x passivation of silicon surfaces are discussed.

Source record↗

Induction of immune-related gene expression by seminal exosomes in the porcine endometrium

Highlights: • Exosomes are present in sperm-rich fraction of porcine seminal plasma (SP). • Exosomes present in SP regulate immune-related genes in the porcine uterus. • Seminal exosomes may pave the way for sperm and fertilized egg transport. Seminal plasma (SP) is considered as a vehicle to carry sperm into female reproductive tract, of which functions have not been completely understood. This study aimed to identify the function of seminal exosomes on porcine endometrium. Exosomes were isolated from the sperm-rich fraction of boar semen and were confirmed by the expression of exosome marker HSP70 and size distribution using nano-sight tracking analysis. Porcine endometrial epithelial cells (EECs) were then treated with seminal exosomes, and RNA extracted were subjected to global expression analysis. Transcripts related to “immune response”, “inflammatory response” and their associated signaling pathways were up-regulated in EECs treated with seminal exosome, whereas those associated with “steroid biosynthesis”, “metabolic pathways” and “T cell differentiation” were down-regulated. The decrease in PMVK, SC5D, INSIG1, HSD17B7, NSDHL, HMGCR, SQLE and FDFT1, and increase in CCL20, TNFSF15, AMCFII, CXCL2 and CXCL8 were also found in the endometrium from the naturally mated pigs. Moreover, changes in exosome-induced CYP24A1, EBP, CCL20, AMCFII and IL1A expression were not regulated by the exosome removed SP. These observations indicated that exosomes present in SP are involved in the immune-related gene regulation in the uterus, which could pave the passage for sperm and possibly fertilized eggs.

60 APPLIED LIFE SCIENCES↗

Spliceosome-associated factor CTNNBL1 promotes proliferation and invasion in ovarian cancer

Ovarian cancer is the most lethal gynecologic malignancy and the molecular pathogenesis of high-grade serous ovarian carcinoma has not been completely characterized. Numerous studies have shown that altered splicing patterns and splicing factors were found to contribute to tumor development and progression. In this study, we demonstrated that spliceosome-associated factor CTNNBL1 was significantly upregulated in high-grade serous ovarian carcinoma, the elevated level of CTNNBL1 indicates poor prognosis in patients with high-grade serous ovarian carcinoma. Functional characterization revealed that CTNNBL1 promoted the proliferation and invasion of ovarian cancer cells in vitro. Furthermore, through transcriptome analysis, we found CTNNBL1 regulates multiple splicing events and gene expression in ovarian cancer cells. Importantly, we identified IFI16 and FOXM1 splicing was regulated by CTNNBL1. To our knowledge, this is the first study exploring the expression, functional roles and regulated splicing events of CTNNBL1 in ovarian cancer. - Highlights: • CTNNBL1 was upregulated in high-grade serous ovarian carcinoma and associated with poor prognosis. • CTNNBL1 promoted the proliferation and invasion of ovarian cancer cells in vitro. • Transcriptome analysis showed CTNNBL1 regulates multiple splicing events and gene expression in ovarian cancer cells. • FOXM1and IFI16 splicing were regulated by CTNNBL1.

60 APPLIED LIFE SCIENCES↗

M-I-S solar cell - Theory and experimental results

The paper presents an operating-mode analysis of an MIS solar cell and discusses the advantages which can arise as a result of the use of transport control, field shaping (increased n factor), and zero bias barrier height modification. It is noted that for an n-type semiconductor, it is relatively easy to obtain an enhanced n factor using acceptor-like states without an increase in diode saturation current, the converse being true for p-type semiconductors. Several MIS configurations are examined: an acceptor-like, localized state configuration producing field shaping and no change in diode saturation current, and acceptor-like localized configurations producing field shaping, with a decrease of diode saturation current, in one case, and an increase in the other.

Childs, R.↗

p{sup +}-doping analysis of laser fired contacts for silicon solar cells by Kelvin probe force microscopy

Local rear contacts for silicon passivated emitter and rear contact solar cells can be established by point-wise treating an Al layer with laser radiation and thereby establishing an electrical contact between Al and Si bulk through the dielectric passivation layer. In this laser fired contacts (LFC) process, Al can establish a few μm thick p{sup +}-doped Si region below the metal/Si interface and forms in this way a local back surface field which reduces carrier recombination at the contacts. In this work, the applicability of Kelvin probe force microscopy (KPFM) to the investigation of LFCs considering the p{sup +}-doping distribution is demonstrated. The method is based on atomic force microscopy and enables the evaluation of the lateral 2D Fermi-level characteristics at sub-micrometer resolution. The distribution of the electrical potential and therefore the local hole concentration in and around the laser fired region can be measured. KPFM is performed on mechanically polished cross-sections of p{sup +}-doped Si regions formed by the LFC process. The sample preparation is of great importance because the KPFM signal is very surface sensitive. Furthermore, the measurement is responsive to sample illumination and the height of the applied voltage between tip and sample. With other measurement techniques like micro-Raman spectroscopy, electrochemical capacitance-voltage, and energy dispersive X-ray analysis, a high local hole concentration in the range of 10{sup 19 }cm{sup −3} is demonstrated in the laser fired region. This provides, in combination with the high spatial resolution of the doping distribution measured by KPFM, a promising approach for microscopic understanding and further optimization of the LFC process.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Genetic Analysis of Ca 2+ Priming in Arabidopsis Guard Cell Stomatal Closure in Response to the Drought Hormone Abscisic Acid

A primary objective of modern agriculture and biofuel production is to utilize arable land to its fullest potential. However, sub-optimal growing conditions—arising from abiotic stresses such as drought, soil salinity, low humidity, cold, and heat—reduce crop yield and quality. Optimal yield under both stressed and non-stressed conditions requires the plant to activate coping mechanisms at a level commensurate with the severity of the drought stress. The osmotic sensors and associated regulatory mechanisms that initiate drought- and salt-tolerance responses in plants are largely unknown. This research aimed to identify and characterize these initial sensory components.

59 BASIC BIOLOGICAL SCIENCES↗

Multi-Objective Boundary Analysis of Discrete and Integrated SiC FET Modular Non-inverting Buck and Boost Converters for Fuel Cell EVs

This paper presents a multi-objective analysis of discrete and integrated SiC FET-based non-inverting buck-boost converter modules for modular fuel cell electric vehicle (EV) systems. Two converter ratings, 60 kW and 90 kW, are evaluated for both implementations, scalable up to 420 kW and 450 kW, respectively. Performance is assessed across efficiency, volumetric and gravimetric power density, cost, thermal stress, and estimated lifetime, where lifetime is derived from SiC FET B10 power-cycling data and junction temperature variations at rated power. A normalized overall performance index combined with a Pareto-boundary framework is used to identify configurations that optimally balance competing objectives. Results show that most configurations lie on the Pareto front, providing balanced trade-offs, while certain high-power discrete (90 kW at 450 kW) and integrated (60 kW at 180−420 kW) configurations are dominated. In general, discrete modules are more favorable for lower-power modular systems due to higher power density and lower cost, whereas integrated modules become more advantageous at higher power levels due to improved thermal behavior and longer lifetime. These findings provide practical design guidance for scalable fuel cell converter architectures and highlight the importance of system-level trade-offs in modular power electronics design.

Asa, Erdem [ORNL] (ORCID:0000000190884812)↗

Mass Spectral Imaging to Map Plant–Microbe Interactions

Plant–microbe interactions are of rising interest in plant sustainability, biomass production, plant biology, and systems biology. These interactions have been a challenge to detect until recent advancements in mass spectrometry imaging. Plants and microbes interact in four main regions within the plant, the rhizosphere, endosphere, phyllosphere, and spermosphere. This mini review covers the challenges within investigations of plant and microbe interactions. We highlight the importance of sample preparation and comparisons among time-of-flight secondary ion mass spectroscopy (ToF-SIMS), matrix-assisted laser desorption/ionization (MALDI), laser desorption ionization (LDI/LDPI), and desorption electrospray ionization (DESI) techniques used for the analysis of these interactions. Using mass spectral imaging (MSI) to study plants and microbes offers advantages in understanding microbe and host interactions at the molecular level with single-cell and community communication information. More research utilizing MSI has emerged in the past several years. We first introduce the principles of major MSI techniques that have been employed in the research of microorganisms. An overview of proper sample preparation methods is offered as a prerequisite for successful MSI analysis. Traditionally, dried or cryogenically prepared, frozen samples have been used; however, they do not provide a true representation of the bacterial biofilms compared to living cell analysis and chemical imaging. New developments such as microfluidic devices that can be used under a vacuum are highly desirable for the application of MSI techniques, such as ToF-SIMS, because they have a subcellular spatial resolution to map and image plant and microbe interactions, including the potential to elucidate metabolic pathways and cell-to-cell interactions. Promising results due to recent MSI advancements in the past five years are selected and highlighted. The latest developments utilizing machine learning are captured as an important outlook for maximal output using MSI to study microorganisms.

59 BASIC BIOLOGICAL SCIENCES↗

SU-F-J-207: Non-Small Cell Lung Cancer Patient Survival Prediction with Quantitative Tumor Textures Analysis in Baseline CT

Purpose: Chemo-radiation therapy (CRT) is widely used in treating patients with locally advanced non-small cell lung cancer (NSCLC). Determination of the likelihood of patient response to treatment and optimization of treatment regime is of clinical significance. Up to date, no imaging biomarker has reliably correlated to NSCLC patient survival rate. This pilot study is to extract CT texture information from tumor regions for patient survival prediction. Methods: Thirteen patients with stage II-III NSCLC were treated using CRT with a median dose of 6210 cGy. Non-contrast-enhanced CT images were acquired for treatment planning and retrospectively collected for this study. Texture analysis was applied in segmented tumor regions using the Local Binary Pattern method (LBP). By comparing its HU with neighboring voxels, the LBPs of a voxel were measured in multiple scales with different group radiuses and numbers of neighbors. The LBP histograms formed a multi-dimensional texture vector for each patient, which was then used to establish and test a Support Vector Machine (SVM) model to predict patients’ one year survival. The leave-one-out cross validation strategy was used recursively to enlarge the training set and derive a reliable predictor. The predictions were compared with the true clinical outcomes. Results: A 10-dimensional LBP histogram was extracted from 3D segmented tumor region for each of the 13 patients. Using the SVM model with the leave-one-out strategy, only 1 out of 13 patients was misclassified. The experiments showed an accuracy of 93%, sensitivity of 100%, and specificity of 86%. Conclusion: Within the framework of a Support Vector Machine based model, the Local Binary Pattern method is able to extract a quantitative imaging biomarker in the prediction of NSCLC patient survival. More patients are to be included in the study.

60 APPLIED LIFE SCIENCES↗

Regional epithelial cell diversity in the small intestine of pigs

Abstract Understanding regional distribution and specialization of small intestinal epithelial cells is crucial for developing methods to control appetite, stress, and nutrient uptake in swine. To establish a better understanding of specific epithelial cells found across different regions of the small intestine in pigs, we utilized single-cell RNA sequencing (scRNA-seq) to recover and analyze epithelial cells from duodenum, jejunum, and ileum. Cells identified included crypt cells, enterocytes, BEST4 enterocytes, goblet cells, and enteroendocrine (EE) cells. EE cells were divided into two subsets based on the level of expression of the EE lineage commitment gene, NEUROD1. NEUROD1hi EE cells had minimal expression of hormone-encoding genes and were dissimilar to EE cells in humans and mice, indicating a subset of EE cells unique to pigs. Recently discovered BEST4 enterocytes were detected in both crypts and villi throughout the small intestine via in situ staining, unlike in humans, where BEST4 enterocytes are found only in small intestinal villi. Proximal-to-distal gradients of expression were noted for hormone-encoding genes in EE cells and nutrient transport genes in enterocytes via scRNA-seq, demonstrating regional specialization. Regional gene expression in EE cells and enterocytes was validated via quantitative PCR (qPCR) analysis of RNA isolated from epithelial cells of different small intestinal locations. Though many genes had similar patterns of regional expression when assessed by qPCR of total epithelial cells, some regional expression was only detected via scRNA-seq, highlighting advantages of scRNA-seq to deconvolute cell type-specific regional gene expression when compared to analysis of bulk samples. Overall, results provide new information on regional localization and transcriptional profiles of epithelial cells in the pig small intestine.

Agriculture↗