Showing papers by "National Cheng Kung University published in 2012"

Guidelines for the use and interpretation of assays for monitoring autophagy

Abstract: In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.

Afatinib versus placebo for patients with advanced, metastatic non-small-cell lung cancer after failure of erlotinib, gefitinib, or both, and one or two lines of chemotherapy (LUX-Lung 1): a phase 2b/3 randomised trial

Abstract: Summary Background Afatinib, an irreversible ErbB-family blocker, has shown preclinical activity when tested in EGFR mutant models with mutations that confer resistance to EGFR tyrosine-kinase inhibitors. We aimed to assess its efficacy in patients with advanced lung adenocarcinoma with previous treatment failure on EGFR tyrosine-kinase inhibitors. Methods In this phase 2b/3 trial, we enrolled patients with stage IIIB or IV adenocarcinoma and an Eastern Cooperative Oncology Group performance (ECOG) performance score of 0–2 who had received one or two previous chemotherapy regimens and had disease progression after at least 12 weeks of treatment with erlotinib or gefitinib. We used a computer-generated sequence to randomly allocate patients (2:1) to either afatinib (50 mg per day) or placebo; all patients received best supportive care. Randomisation was done in blocks of three and was stratified by sex and baseline ECOG performance status (0–1 vs 2). Investigators, patients, and the trial sponsor were masked to treatment assignment. The primary endpoint was overall survival (from date of randomisation to death), analysed on an intention-to-treat basis. This study is registered with ClinicalTrials.gov, number NCT00656136. Findings Between May 26, 2008, and Sept 21, 2009, we identified 697 patients, 585 of whom were randomly allocated to treatment (390 to afatinib, 195 to placebo). Median overall survival was 10·8 months (95% CI 10·0–12·0) in the afatinib group and 12·0 months (10·2–14·3) in the placebo group (hazard ratio 1·08, 95% CI 0·86–1·35; p=0·74). Median progression-free survival was longer in the afatinib group (3·3 months, 95% CI 2·79–4·40) than it was in the placebo group (1·1 months, 0·95–1·68; hazard ratio 0·38, 95% CI 0·31–0·48; p Interpretation Although we recorded no benefit in terms of overall survival with afatinib (which might have been affected by cancer treatments given after progression in both groups), our findings for progression-free survival and response to treatment suggest that afatinib could be of some benefit to patients with advanced lung adenocarcinoma who have failed at least 12 weeks of previous EGFR tyrosine-kinase inhibitor treatment. Funding Boehringer Ingelheim Inc.

Digital storytelling for enhancing student academic achievement, critical thinking, and learning motivation: A year-long experimental study

Abstract: The purpose of this study was to explore the impact of Digital storytelling (DST) on the academic achievement, critical thinking, and learning motivation of senior high school students learning English as a foreign language. The one-year study adopted a pretest and posttest quasi-experimental design involving 110 10th grade students in two English classes. The independent variable was information technology-integrated instruction (ITII) on two different levels - lecture-type ITII (comparison group) and DST (experimental group). Both quantitative and qualitative data were collected, including English achievement and critical thinking scores, questionnaire responses for learning motivation, as well as recordings of student and teacher interviews for evaluating the effectiveness of DST in learning. Descriptive analysis, analysis of covariance (ANCOVA), multivariate analysis of covariance (MANCOVA), and qualitative content analysis was used for evaluating the obtained data. Our findings indicate that DST participants performed significantly better than lecture-type ITII participants in terms of English achievement, critical thinking, and learning motivation. Interview results highlight the important educational value of DST, as both the instructor and students reported that DST increased students' understanding of course content, willingness to explore, and ability to think critically, factors which are important in preparing students for an ever-changing 21st century.

Mitochondrial fission contributes to mitochondrial dysfunction and insulin resistance in skeletal muscle.

Abstract: Mitochondrial dysfunction in skeletal muscle has been implicated in the development of insulin resistance and type 2 diabetes. Considering the importance of mitochondrial dynamics in mitochondrial and cellular functions, we hypothesized that obesity and excess energy intake shift the balance of mitochondrial dynamics, further contributing to mitochondrial dysfunction and metabolic deterioration in skeletal muscle. First, we revealed that excess palmitate (PA), but not hyperglycemia, hyperinsulinemia, or elevated tumor necrosis factor alpha, induced mitochondrial fragmentation and increased mitochondrion-associated Drp1 and Fis1 in differentiated C2C12 muscle cells. This fragmentation was associated with increased oxidative stress, mitochondrial depolarization, loss of ATP production, and reduced insulin-stimulated glucose uptake. Both genetic and pharmacological inhibition of Drp1 attenuated PA-induced mitochondrial fragmentation, mitochondrial depolarization, and insulin resistance in C2C12 cells. Furthermore, we found smaller and shorter mitochondria and increased mitochondrial fission machinery in the skeletal muscle of mice with genetic obesity and those with diet-induced obesity. Inhibition of mitochondrial fission improved the muscle insulin signaling and systemic insulin sensitivity of obese mice. Our findings indicated that aberrant mitochondrial fission is causally associated with mitochondrial dysfunction and insulin resistance in skeletal muscle. Thus, disruption of mitochondrial dynamics may underlie the pathogenesis of muscle insulin resistance in obesity and type 2 diabetes.

A Novel Class of Strain Gauges Based on Layered Percolative Films of 2D Materials

Abstract: Here we report on the fabrication and characterization of a novel type of strain gauge based on percolative networks of 2D materials. The high sensitivity of the percolative carrier transport to strain induced morphology changes was exploited in strain sensors that can be produced from a wide variety of materials. Highly reliable and sensitive graphene-based thin film strain gauges were produced from solution processed graphene flakes by spray deposition. Control of the gauge sensitivity could be exerted through deposition-induced changes to the film morphology. This exceptional property was explained through modeling of the strain induced changes to the flake–flake overlap for different percolation networks. The ability to directly deposit strain gauges on complex-shaped and transparent surfaces was presented. The demonstrated scalable fabrication, superior sensitivity over conventional sensors, and unique properties of the described strain gauges have the potential to improve existing technology and ope...

Afatinib for patients with lung adenocarcinoma and epidermal growth factor receptor mutations (LUX-Lung 2): A phase 2 trial

Abstract: Summary Background Afatinib is an irreversible ErbB-family blocker with preclinical activity in non-small-cell lung cancer (NSCLC) with EGFR mutations. We aimed to assess the efficacy of afatinib in patients with lung adenocarcinoma and EGFR mutations. Methods In this phase 2 study, we enrolled patients from 30 centres in Taiwan and the USA with lung adenocarcinoma (stage IIIb with pleural effusion or stage IV) with EGFR mutations, who had no more than one previous chemotherapy regimen for advanced disease, an Eastern Cooperative Oncology Group performance status of 0–2, and no previous treatment with EGFR tyrosine-kinase inhibitors. We tested two afatinib starting doses: 50 mg daily and subsequently 40 mg daily, introduced to establish whether tolerability could be improved with retention of anti-tumour activity. The primary endpoint was the proportion of patients with a confirmed objective response (complete response or partial response), on the basis of Response Evaluation Criteria in Solid Tumors 1.0 (independent review). This study is registered with ClinicalTrials.gov, number NCT00525148. Findings 129 patients were treated with afatinib, 99 with a starting dose of 50 mg and 30 with a starting dose of 40 mg. 79 (61%) of 129 patients had an objective response (two complete responses, 77 partial responses). 70 (66%) of the 106 patients with the two common activating EGFR mutations (deletion 19 or L858R) had an objective response, as did nine (39%) of 23 patients with less common mutations. Similar proportions of patients had an objective response when analysed by starting dose (18 [60%] of 30 patients at 40 mg vs 61 [62%] of 99 patients at 50 mg). Of the two most common adverse events (diarrhoea and rash or acne), grade 3 events were more common in patients receiving a 50 mg starting dose (22 [22%] of 99 patients for diarrhoea and 28 [28%] of 99 patients for rash or acne) than they were in those receiving a 40 mg starting dose (two [7%] of 30 patients for both diarrhoea and rash or acne); possibly treatment-related serious adverse events were also less common in patients receiving a 40 mg starting dose (two of 30 patients vs 14 of 99 patients). We recorded one possibly drug-related death (interstitial lung disease). Interpretation Afatinib shows activity in the treatment of patients with advanced lung adenocarcinoma with EGFR mutations, especially in patients with deletion 19 or L858R mutations. The efficacy of afatinib 40 mg should be compared with chemotherapy or other EGFR tyrosine-kinase inhibitors in EGFR-mutation-positive NSCLC. Funding Boehringer Ingelheim Inc.

Autophagy activators rescue and alleviate pathogenesis of a mouse model with proteinopathies of the TAR DNA-binding protein 43

Abstract: TDP-43 is a multifunctional DNA/RNA-binding protein that has been identified as the major component of the cytoplasmic ubiquitin (+) inclusions (UBIs) in diseased cells of frontotemporal lobar dementia (FTLD-U) and amyotrophic lateral sclerosis (ALS). Unfortunately, effective drugs for these neurodegenerative diseases are yet to be developed. We have tested the therapeutic potential of rapamycin, an inhibitor of the mammalian target of rapamycin (mTOR) and three other autophagy activators (spermidine, carbamazepine, and tamoxifen) in a FTLD-U mouse model with TDP-43 proteinopathies. Rapamycin treatment has been reported to be beneficial in some animal models of neurodegenerative diseases but not others. Furthermore, the effects of rapamycin treatment in FTLD-U have not been investigated. We show that rapamycin treatment effectively rescues the learning/memory impairment of these mice at 3 mo of age, and it significantly slows down the age-dependent loss of their motor function. These behavioral improvements upon rapamycin treatment are accompanied by a decreased level of caspase-3 and a reduction of neuron loss in the forebrain of FTLD-U mice. Furthermore, the number of cells with cytosolic TDP-43 (+) inclusions and the amounts of full-length TDP-43 as well as its cleavage products (35 kDa and 25 kDa) in the urea-soluble fraction of the cellular extract are significantly decreased upon rapamycin treatment. These changes in TDP-43 metabolism are accompanied by rapamycin-induced decreases in mTOR-regulated phospho-p70 S6 kinase (P-p70) and the p62 protein, as well as increases in the autophagic marker LC3. Finally, rapamycin as well as spermidine, carbamazepine, and tamoxifen could also rescue the motor dysfunction of 7-mo-old FTLD-U mice. These data suggest that autophagy activation is a potentially useful route for the therapy of neurodegenerative diseases with TDP-43 proteinopathies.

High-performance quantum dot-sensitized solar cells based on sensitization with CuInS2 quantum dots/CdS heterostructure

Abstract: A high-performance quantum dot-sensitized solar cell (QDSSC) is reported, which consists of a TiO2/CuInS2-QDs/CdS/ZnS photoanode, a polysulfide electrolyte, and a CuS counter electrode. The sensitization process involves attaching presynthesized CuInS2 QDs (3.5 nm) to a TiO2 substrate with a bifunctional linker, followed by coating CdS with successive ionic layer adsorption and reaction (SILAR) and ZnS as the last SILAR layer for passivation. This process constructs a sensitizing layer that comprises CdS nanocrystals, closely packed around the earlier-linked CuInS2 QDs, which serve as the pillars of the layer. The CuS counter electrode, prepared via successive ionic solution coating and reaction, has a small charge transfer resistance in the polysulfide electrolyte. The QDSSC exhibits a short-circuit photocurrent (Jsc) of 16.9 mA cm−2, an open-circuit photovoltage (Voc) of 0.56 V, a fill factor of 0.45, and a conversion efficiency of 4.2% under one-sun illumination. The heterojunction between the CuInS2 QDs and CdS extends both the optical absorption and incident photon conversion efficiency (IPCE) spectra of the cell to a longer wavelength of approximately 800 nm, and provides an IPCE of nearly 80% at 510 nm. The high TiO2 surface coverage of the sensitizers suppresses recombination of the photogenerated electrons. This results in a longer lifetime for the electrons, and therefore, the high Voc value. The notably high Jsc and Voc values demonstrate that this sensitization strategy, which exploits the quantum confinement reduction and other synergistic effects of the CuInS2-QDs/CdS/ZnS heterostructure, can potentially outperform those of other QDSSCs.

Plasmon Inducing Effects for Enhanced Photoelectrochemical Water Splitting: X-ray Absorption Approach to Electronic Structures

Abstract: Artificial photosynthesis using semiconductors has been investigated for more than three decades for the purpose of transferring solar energy into chemical fuels. Numerous studies have revealed that the introduction of plasmonic materials into photochemical reaction can substantially enhance the photo response to the solar splitting of water. Until recently, few systematic studies have provided clear evidence concerning how plasmon excitation and which factor dominates the solar splitting of water in photovoltaic devices. This work demonstrates the effects of plasmons upon an Au nanostructure–ZnO nanorods array as a photoanode. Several strategies have been successfully adopted to reveal the mutually independent contributions of various plasmonic effects under solar irradiation. These have clarified that the coupling of hot electrons that are formed by plasmons and the electromagnetic field can effectively increase the probability of a photochemical reaction in the splitting of water. These findings suppor...

Novel High Step-Up DC–DC Converter With Coupled-Inductor and Switched-Capacitor Techniques

Abstract: A novel high step-up dc-dc converter with coupled-inductor and switched-capacitor techniques is proposed in this paper. The capacitors are charged in parallel and are discharged in series by the coupled inductor, stacking on the output capacitor. Thus, the proposed converter can achieve high step-up voltage gain with appropriate duty ratio. Besides, the voltage spike on the main switch can be clamped. Therefore, low on-state resistance RDS(ON) of the main switch can be adopted to reduce the conduction loss. The efficiency can be improved. The operating principle and steady-state analyses are discussed in detail. Finally, a prototype circuit with 24-V input voltage, 400-V output voltage, and 200-W output power is implemented in the laboratory. Experiment results confirm the analysis and advantages of the proposed converter.

Genome-wide association analysis identifies new lung cancer susceptibility loci in never-smoking women in Asia.

Abstract: To identify common genetic variants that contribute to lung cancer susceptibility, we conducted a multistage genome-wide association study of lung cancer in Asian women who never smoked. We scanned 5,510 never-smoking female lung cancer cases and 4,544 controls drawn from 14 studies from mainland China, South Korea, Japan, Singapore, Taiwan and Hong Kong. We genotyped the most promising variants (associated at P < 5 × 10(-6)) in an additional 1,099 cases and 2,913 controls. We identified three new susceptibility loci at 10q25.2 (rs7086803, P = 3.54 × 10(-18)), 6q22.2 (rs9387478, P = 4.14 × 10(-10)) and 6p21.32 (rs2395185, P = 9.51 × 10(-9)). We also confirmed associations reported for loci at 5p15.33 and 3q28 and a recently reported finding at 17q24.3. We observed no evidence of association for lung cancer at 15q25 in never-smoking women in Asia, providing strong evidence that this locus is not associated with lung cancer independent of smoking.

General non-Markovian dynamics of open quantum systems.

Abstract: We present a general theory of non-Markovian dynamics for open systems of noninteracting fermions (bosons) linearly coupled to thermal environments of noninteracting fermions (bosons). We explore the non-Markovian dynamics by connecting the exact master equations with the nonequilibirum Green's functions. Environmental backactions are fully taken into account. The non-Markovian dynamics consists of nonexponential decays and dissipationless oscillations. Nonexponential decays are induced by the discontinuity in the imaginary part of the self-energy corrections. Dissipationless oscillations arise from band gaps or the finite band structure of spectral densities. The exact analytic solutions for various non-Markovian thermal environments show that non-Markovian dynamics can be largely understood from the environmental-modified spectra of open systems.

A ubiquitous English vocabulary learning system: Evidence of active/passive attitudes vs. usefulness/ease-of-use

Abstract: English vocabulary learning and ubiquitous learning have separately received considerable attention in recent years. However, research on English vocabulary learning in ubiquitous learning contexts has been less studied. In this study, we develop a ubiquitous English vocabulary learning (UEVL) system to assist students in experiencing a systematic vocabulary learning process in which ubiquitous technology is used to develop the system, and video clips are used as the material. Afterward, the technology acceptance model and partial least squares approach are used to explore students' perspectives on the UEVL system. The results indicate that (1) both the system characteristics and the material characteristics of the UEVL system positively and significantly influence the perspectives of all students on the system; (2) the active students are interested in perceived usefulness; (3) the passive students are interested in perceived ease of use.

Automatic Stage Scoring of Single-Channel Sleep EEG by Using Multiscale Entropy and Autoregressive Models

Abstract: In this paper, we propose an automatic sleep-scoring method combining multiscale entropy (MSE) and autoregressive (AR) models for single-channel EEG and to assess the performance of the method comparatively with manual scoring based on full polysomnograms. This is the first time that MSE has ever been applied to sleep scoring. All-night polysomnograms from 20 healthy individuals were scored using the Rechtschaffen and Kales rules. The developed method analyzed the EEG signals of C3-A2 for sleep staging. The results of automatic and manual scorings were compared on an epoch-by-epoch basis. A total of 8480 30-s sleep EEG epochs were measured and used for performance evaluation. The epoch-by-epoch comparison was made by classifying the EEG epochs into five states (Wake/REM/S1/S2/SWS) by the proposed method and manual scoring. The overall sensitivity and kappa coefficient of MSE alone are 76.9% and 0.65, respectively. Moreover, the overall sensitivity and kappa coefficient of our proposed method of integrating MSE, AR models, and a smoothing process can reach the sensitivity level of 88.1% and 0.81, respectively. Our results show that MSE is a useful and representative feature for sleep staging. It has high accuracy and good home-care applicability because a single EEG channel is used for sleep staging.

DNAzyme targeting c-jun suppresses skin cancer growth

Abstract: Worldwide, one in three cancers is skin-related, with increasing incidence in many populations Here, we demonstrate the capacity of a DNAzyme-targeting c-jun mRNA, Dz13, to inhibit growth of two common skin cancer types-basal cell and squamous cell carcinomas-in a therapeutic setting with established tumors Dz13 inhibited tumor growth in both immunodeficient and immunocompetent syngeneic mice and reduced lung nodule formation in a model of metastasis In addition, Dz13 suppressed neovascularization in tumor-bearing mice and zebrafish and increased apoptosis of tumor cells Dz13 inhibition of tumor growth, which required an intact catalytic domain, was due in part to the induction of tumor immunity In a series of good laboratory practice-compliant toxicology studies in cynomolgus monkeys, minipigs, and rodents, the DNAzyme was found to be safe and well tolerated It also did not interfere in more than 70 physiologically relevant in vitro bioassays, suggesting a reduced propensity for off-target effects If these findings hold true in clinical trials, Dz13 may provide a safe, effective therapy for human skin cancer

Witnessing Quantum Coherence: from solid-state to biological systems

Abstract: Quantum coherence is one of the primary non-classical features of quantum systems. While protocols such as the Leggett-Garg inequality (LGI) and quantum tomography can be used to test for the existence of quantum coherence and dynamics in a given system, unambiguously detecting inherent “quantumness” still faces serious obstacles in terms of experimental feasibility and efficiency, particularly in complex systems. Here we introduce two “quantum witnesses” to efficiently verify quantum coherence and dynamics in the time domain, without the expense and burden of non-invasive measurements or full tomographic processes. Using several physical examples, including quantum transport in solid-state nanostructures and in biological organisms, we show that these quantum witnesses are robust and have a much finer resolution in their detection window than the LGI has. These robust quantum indicators may assist in reducing the experimental overhead in unambiguously verifying quantum coherence in complex systems.

Fast and slow rates of naphthalene sorption to biochars produced at different temperatures.

Abstract: This study investigated the sorption kinetics of a model solute (naphthalene) with a series of biochars prepared from a pine wood at 150–700 °C (referred as PW100–PW700) to probe the effect of the degree of carbonization of a biochar. The samples were characterized by the elemental compositions, thermal gravimetric analyses, Fourier transform IR spectroscopy, scanning electron microscopy, Brunauer–Emmett–Teller-N2 surface areas (SA), and pore size distributions. Naphthalene exhibited a fast rate of sorption to PW150 owning a high oxygen content and a small SA, due supposedly to the solute partition into a swollen well-hydrated uncarbonized organic matter of PW150. The partial removal of polar-group contents in PW250/PW350, which increased the compactness of the partition medium, decreased the diffusion of the solute into the partition phase to result in a slow sorption rate. With PW500 and PW700 displaying low oxygen contents and high SA, the solute sorption rates were fast, attributed to the near exhaust...