NMR studies of the relationship between the changes of membrane lipids and the cisplatin-resistance of A549/DDP cells

Changes of membrane lipids in cisplatin-sensitive A549 and cisplatin-resistant A549/DDP cells during the apoptotic process induced by a clinical dose of cisplatin (30 μM) were detected by (1)H and (31)P-NMR spectroscopy and by membrane fluidity measurement. The apoptotic phenotypes of the two cell lines were monitored with flow cytometry. The assays of apoptosis showed that significant apoptotic characteristics of the A549 cells were induced when the cells were cultured for 24 hours after treatment with cisplatin, while no apoptotic characteristic could be detected for the resistant A549/DDP cells even after 48 hours. The results of (1)H-NMR spectroscopy demonstrated that the CH(2)/CH(3 )and Glu/Ct ratios of the membrane of A549 cells increased significantly, but those in A549/DDP cell membranes decreased. In addition, the Chol/CH(3 )and Eth/Ct ratios decreased for the former but increased for the latter cells under the same conditions. (31)P-NMR spectroscopy indicated levels of phosphomonoesters (PME) and ATP decreased in A549 but increased in A549/DDP cells after being treated with cisplatin. These results were supported with the data obtained from (1)H-NMR measurements. The results clearly indicated that components and properties of membrane phospholipids of the two cell lines were significantly different during the apoptotic process when they were treated with a clinical dose of cisplatin. Plasma membrane fluidity changes during cisplatin treatment as detected with the fluorescence probe TMA-DPH also indicate marked difference between the two cell lines. We provided evidence that there are significant differences in plasma membrane changes during treatment of cisplatin sensitive A549 and resistant A549/DDP cells

Modeling of Performance Creative Evaluation Driven by Multimodal Affective Data

Performance creative evaluation can be achieved through affective data, and the use of affective featuresto evaluate performance creative is a new research trend. This paper proposes a “Performance Creative—Multimodal Affective (PC-MulAff)” model based on the multimodal affective features for performance creative evaluation. The multimedia data acquisition equipment is used to collect the physiological data of the audience, including the multimodal affective data such as the facial expression, heart rate and eye movement. Calculate affective features of multimodal data combined with director annotation, and defined “Performance Creative—Affective Acceptance (PC-Acc)” based on multimodal affective features to evaluate the quality of performance creative. This paper verifies the PC-MulAff model on different performance data sets. The experimental results show that the PC-MulAff model shows high evaluation quality in different performance forms. In the creative evaluation of dance performance, the accuracy of the model is 7.44% and 13.95% higher than that of the single textual and single video evaluation

Crystal Structure of the N-Acetylmannosamine Kinase Domain of GNE

UDP-GlcNAc 2-epimerase/ManNAc 6-kinase, GNE, is a bi-functional enzyme that plays a key role in sialic acid biosynthesis. Mutations of the GNE protein cause sialurea or autosomal recessive inclusion body myopathy/Nonaka myopathy. GNE is the only human protein that contains a kinase domain belonging to the ROK (repressor, ORF, kinase) family.We solved the structure of the GNE kinase domain in the ligand-free state. The protein exists predominantly as a dimer in solution, with small populations of monomer and higher-order oligomer in equilibrium with the dimer. Crystal packing analysis reveals the existence of a crystallographic hexamer, and that the kinase domain dimerizes through the C-lobe subdomain. Mapping of disease-related missense mutations onto the kinase domain structure revealed that the mutation sites could be classified into four different groups based on the location - dimer interface, interlobar helices, protein surface, or within other secondary structural elements.The crystal structure of the kinase domain of GNE provides a structural basis for understanding disease-causing mutations and a model of hexameric wild type full length enzyme.This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3D representations and animated transitions. Please note that a web plugin is required to access this enhanced functionality. Instructions for the installation and use of the web plugin are available in Text S1

Multiple roles of PPAR alpha in brown adipose tissue under constitutive and cold conditions

Peroxisome proliferator-activated receptor alpha (PPAR alpha) is a member of the nuclear receptor family, regulating fatty acid degradation in many organs. Two-dimensional SDS-PAGE of brown adipose tissue (BAT) from PPAR alpha-null mice produced a higher-density spot. Proteomic analysis indicated that the protein was pyruvate dehydrogenase beta (PDH beta). To observe PDH beta regulation in BAT, the organ was stimulated by long-term cold exposure, and the activities of associated enzymes were investigated. Histological and biochemical analyses of BAT showed a significant decrease in the triglyceride content in wild-type mice and some degree of decrease in PPAR alpha-null mice on cold exposure. Analyses of molecules related to glucose metabolism showed that the expression of PDH beta is under PPAR alpha-specific regulation, and that glucose degradation ability may decrease on cold exposure. In contrast, analyses of molecules related to fatty acid metabolism showed that numerous PPAR alpha/gamma target molecules are induced on cold exposure, and that fatty acid degradation ability in wild-type mice is markedly enhanced and also increases to same degree in PPAR alpha-null mice on cold exposure. Thus, this study proposes novel and multiple roles of PPAR alpha in BAT.ArticleGENES TO CELLS. 15(2):91-100 (2010)journal articl

Crystal Structures of the Tetratricopeptide Repeat Domains of Kinesin Light Chains: Insight into Cargo Recognition Mechanisms

Kinesin-1 transports various cargos along the axon by interacting with the cargos through its light chain subunit. Kinesin light chains (KLC) utilize its tetratricopeptide repeat (TPR) domain to interact with over 10 different cargos. Despite a high sequence identity between their TPR domains (87%), KLC1 and KLC2 isoforms exhibit differential binding properties towards some cargos. We determined the structures of human KLC1 and KLC2 tetratricopeptide repeat (TPR) domains using X-ray crystallography and investigated the different mechanisms by which KLCs interact with their cargos. Using isothermal titration calorimetry, we attributed the specific interaction between KLC1 and JNK-interacting protein 1 (JIP1) cargo to residue N343 in the fourth TRP repeat. Structurally, the N343 residue is adjacent to other asparagines and lysines, creating a positively charged polar patch within the groove of the TPR domain. Whereas, KLC2 with the corresponding residue S328 did not interact with JIP1. Based on these finding, we propose that N343 of KLC1 can form “a carboxylate clamp” with its neighboring asparagine to interact with JIP1, similar to that of HSP70/HSP90 organizing protein-1's (HOP1) interaction with heat shock proteins. For the binding of cargos shared by KLC1 and KLC2, we propose a different site located within the groove but not involving N343. We further propose a third binding site on KLC1 which involves a stretch of polar residues along the inter-TPR loops that may form a network of hydrogen bonds to JIP3 and JIP4. Together, these results provide structural insights into possible mechanisms of interaction between KLC TPR domains and various cargo proteins

The Neutron-Gamma Pulse Shape Discrimination of CLLB Detector

Cs2LiLaBr6: Ce (CLLB) scintillator with the size of Φ 21mm × 25 mm coupled with PMT was used to detect neutron and gamma rays. The pulse shape discrimination (PSD) of neutrons and gamma rays by charge comparison method, the neutrons and gamma rays from AmBe source and fast neutron beam can be separated with figure-of-merit (FOM) values of 0.9 and 1.3, respectively. However, some neutron and gamma rays are difficult to distinguish, so new algorithms need to be investigated to improve the PSD performance of neutron and gamma. Artificial neural networks (ANN) have a very good image recognition capability, thus the ANN model was constructed to discriminate the waveforms of neutron and gamma rays. After ANN model training, the neutron and gamma signals of the CLLB detector were recognized with an accuracy of 98%, and the FOM value of the ANN method was calculated to be 19.4. This result is much higher than the charge comparison method, indicating better discrimination between neutrons and gamma rays with the ANN method

Early tissue and healing responses after maxillary sinus augmentation using horizontal platelet rich fibrin bone blocks.

BACKGROUND The effects of horizontal platelet-rich fibrin (H-PRF) bone block on the healing and immune response during sinus augmentation have not been fully investigated histologically at early time points. METHODS Eighteenth male New Zealand white rabbits underwent bilateral sinus augmentation and were divided into two groups: deproteinized bovine bone mineral (DBBM) alone and H-PRF + DBBM (H-PRF bone block) group. Maxilla samples were collected at 3, 7 and 14 days post sinus augmentation procedures and analyzed using histological staining for the number of inflammatory cells, new blood vessels and evidence for early osteoclast bone turnover/remodeling. Furthermore, the effects of H-PRF bone blocks on the migration of osteoblasts and THP-1 macrophages were evaluated using a Transwell assay in vitro. RESULTS A higher number of immune cells were found in the H-PRF bone block group at 3 and 7 days post-surgery when compared to the DBBM alone group,most notably in the regions close to the mucosal lining and bone plates. Furthermore, a significantly greater number of new blood vessel formations and early signs of osteoclast development were found in the H-PRF bone block group at 14 days. The in vitro transwell assay further confirmed that culture medium from H-PRF bone block markedly promote the migration of osteoblasts and THP-1 macrophages. CONCLUSIONS The findings from this study have shown that H-PRF bone block is capable of increasing early immune cell infiltration leading to the acceleration of neovascularization and speeding the process of bone metabolism in vivo following maxillary sinus grafting with DBBM

The Combination of Human Urinary Kallidinogenase and Mild Hypothermia Protects Adult Rats Against Hypoxic-Ischemic Encephalopathy-Induced Injury by Promoting Angiogenesis and Regeneration

Objectives: Human Urinary Kallidinogenase (HUK) is a tissue kallikrein that plays neuroprotective role in ischemic conditions via different mechanisms. Mild hypothermia (MH) is another robust neuroprotectant that reduces mortality but does not profoundly ameliorate the neurological outcome in hypoxic-ischemic encephalopathy (HIE) patients. However, whether the combination of HUK and MH can be used as a promising neuroprotective treatment in HIE is unknown.Methods: One-hundred and forty-four adult Wistar rats were randomly divided into five groups: Sham, HIE, HUK, MH and a combination of HUK and MH treatment. The HIE rat model was established by right carotid dissection followed by hypoxia aspiration. The survival curve was created within 7 days, and the neurological severity scores (NSS) were assessed at days 0, 1, 3, and 7. Nissl staining, Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL), immunofluorescent staining and western blotting were used to evaluate neuronal survival, apoptosis and necrosis, tight-junction proteins Claudin-1 and Zonula occludens-1 (ZO-1), vascular endothelial growth factor (VEGF), doublecortex (DCX), bradykinin receptor B1 (BDKRB1), BDKRB2 and Ki67 staining.Results: The combined treatment rescued all HIE rats from death and had a best survival curve compared to HIE. The Combination also reduced the NSS scores after HIE at days 7, better than HUK or MH alone. The combination of HUK and MH reserved more cells in Nissl staining and inhibited neuronal apoptosis and necrosis as well as significantly attenuated HIE-induced decreases in claudin-1, ZO-1, cyclin D1 and BDKRB1/B2 in comparison to HUK or MH treatment alone. Moreover, the combined treatment increased the expression of VEGF and DCX as well as the number of Ki67-labeled cells.Conclusions: This study demonstrates that both HUK and MH are neuroprotective after HIE insult; however, the combined therapy with HUK and MH enhanced the efficiency and efficacy of either therapy alone in the treatment of HIE, at least partially by promoting angiogenesis and regeneration and rescuing tight-junction loss. The combination of HUK and MH seems to be a feasible and promising clinical strategy to alleviate cerebral injury following HIE insult.Highlights: -The combination of HUK and MH distinctly reduces neurological dysfunction in HIE rats.-HUK enhances the neuroprotective effects of MH in HIE.-MH attenuates tight-junction disruption, upregulates the BDKR B1/2, DCX and cyclin D1.-The combination of MH and HUK enhances the expressions of MH/HUK mediated-BDKR B1/2, DCX, cyclin D1 and Ki67 positive cells

Research and development of Hg-CEMS flue gas pre-treatment technology

In view of the natural endowment of mercury in fossil fuels and mineral resources, as well as the mercury emission controlling and regulating in the energy and resource utilization processes, the research and development on the mercury continuous emissions monitoring system (Hg-CEMS) with proprietary intellectual property rights is the most important requirement in science and technology in China. As a critical component of the Hg-CEMS system, the flue gas pretreatment system is a core technology that limits the development and application of the Hg-CEMS technology in China. This paper presents a comprehensive review of the research progress in the four key modules of the Hg-CEMS flue gas pretreatment technology, including the dilution sampling, Hg0/Hg2+ separation, Hg2+ reduction and Hg2+ calibration gas generation. Firstly, the dilution sampling techniques are highlighted in their characteristics and principles. The working principles, structures, and design considerations of the critical parameters for both critical hole and thermal dilution injectors are summarized. Emphasis is focused on the connection of numerical simulation with the optimal design methodology for the thermal dilution injector and critical hole. Secondly, regarding the Hg0/Hg2+ separation technology, various partitioning methodologies encompassing the wet absorption separation, physical adsorption separation and chemical adsorption separation are discussed. Particular attention is given to the separation and adsorption efficiency of the advanced dry chemical adsorption techniques involving the novel selective adsorbents of KCl and CaO with their updated theoretical and experimental outcomes under practical operating conditions. Thirdly, in aspect of the Hg2+ reduction technology, a comparative analysis of wet chemical reduction, low-temperature reduction and high-temperature reduction techniques is expatiated. Particular focus is on the active components of solid-state reducing agents, the impact of temperature on the reduction of divalent mercury, and the detrimental influences of flue gas components on the re-oxidation of the reduced elemental mercury in the low-temperature reduction techniques. An in-depth investigation of the impact of filler materials, acidic gas components and acid removal techniques on the reduction of divalent mercury in the high-temperature reduction techniques is highlighted. Finally, facing the technical problems, a solid generation method of the Hg2+ calibration gas is invented and its industrial utilization feasibility is verified in chemistry and experiments. In conjunction with the current status of research and application of the Hg-CEMS technology, the research technical routs, the solutions to some problems and development prospect for the above-mentioned four key modules are put forward