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  1. Meng X, Wen K, Citartan M, Lin Q
    Analyst, 2023 Feb 13;148(4):787-798.
    PMID: 36688616 DOI: 10.1039/d2an01767a
    Aptamers are single-stranded oligonucleotide molecules that bind with high affinity and specificity to a wide range of target molecules. The method of systematic evolution of ligands by exponential enrichment (SELEX) plays an essential role in the isolation of aptamers from a randomized oligonucleotide library. To date, significant modifications and improvements of the SELEX process have been achieved, engendering various forms of SELEX from conventional SELEX to microfluidics-based full-chip SELEX. While full-chip SELEX is generally considered advantageous over conventional SELEX, there has not yet been a conclusive comparison between the methods. Herein, we present a comparative study of three SELEX strategies for aptamer isolation, including those using conventional agarose bead-based partitioning, microfluidic affinity selection, and fully integrated microfluidic affinity selection and PCR amplification. Using immunoglobulin E (IgE) as a model target molecule, we compare these strategies in terms of the time and cost for each step of the SELEX process including affinity selection, amplification, and oligonucleotide conditioning. Target-binding oligonucleotides in the enriched pools are sequenced and compared to assess the relative efficacy of the SELEX strategies. We show that the microfluidic strategies are more time- and cost-efficient than conventional SELEX.
  2. Zhang Q, Sun H, Peng X, Lin Q
    PLoS One, 2023;18(6):e0287227.
    PMID: 37352245 DOI: 10.1371/journal.pone.0287227
    The intention of pro-environmental behavior (PEB) directly affects the sustainable development of protected areas, especially national parks, but few studies have done comparative research on tourist and hiker behaviors. This study explores the intention of tourists' and hikers' pro-environmental behavior based on theory of planned behavior (TPB) and norm activation theory (NAM). Researchers surveyed 454 tourists and 466 hikers in Wuyishan National Park a structural equation modeling data analysis method. The results demonstrate that the TPB and the NAM were accurate in describing for tourists' and hikers' pro-environmental behavior in national park. However, for specific influencing factors, hikers' attitude, awareness of consequences, and assumption of responsibility were significantly different from those of the tourists. This study sheds light on how to better comprehend and advocate for PEB in national parks and proposes different management approaches to improve the PEB of tourists and hikers.
  3. Tian X, Liu X, Xiao Z, Lin Q
    Psychol Res Behav Manag, 2023;16:3151-3169.
    PMID: 37584038 DOI: 10.2147/PRBM.S413738
    INTRODUCTION: This study aims to investigate the impact of COVID-19 on undergraduate students' academic procrastination and negative emotions. It seeks to examine the relationship between negative affect and academic procrastination among university students, considering the mediating role of life autonomy and the moderating effect of a sense of school belonging.

    METHODS: The study sample consisted of 776 university students from the Guangxi Zhuang Autonomous Region, China. Data collection involved the use of the Negative Affect Scale, Academic Procrastination Scale, Life Autonomy Scale, and Sense of School Belonging Scale. SPSS version 26.0 and PROCESS V3.580 to conduct relationship analysis, test the mediation model (model 4) and the moderated mediating model (model 7).

    RESULTS: The findings revealed significant relationships among the variables. Negative emotions were negatively related with life autonomy and sense of school belonging, and positively related with academic procrastination. Academic procrastination showed a negative correlation with both life autonomy and sense of school belonging. Sense of school belonging exhibited a positive relationship with life autonomy. The study also identified life autonomy as a mediator in the relationship between negative emotions and academic procrastination. Additionally, the sense of school belonging moderated the initial segment of the negative emotion-life autonomy-academic procrastination pathway.

    DISCUSSION: This study contributes to a better understanding of the association between negative affect and academic procrastination. It highlights that negative emotions have a direct and indirect influence on academic procrastination through the mediating role of life autonomy. Moreover, the moderating role of sense of school belonging suggests that the relationship between negative affect and life autonomy is stronger when the sense of school belonging is weaker.

  4. Chua LS, Abdullah FI, Lim TK, Lin Q
    Food Chem, 2024 Jan 30;432:137261.
    PMID: 37651783 DOI: 10.1016/j.foodchem.2023.137261
    This study was aimed to extract bioactive peptides from the white and purple flower varieties of Orthosiphon aristatus leaves. The herb is well known for its pharmacological importance, possibly attributed to its plant proteins. Phenol based extraction was used to extract plant proteins, and then hydrolysed by proteolytic enzymes such as trypsin (serine protease) and pepsin (aspartic protease). MS/MS analysis revealed that 145 and 125 proteins were detected from the white and purple flower varieties, respectively. Trypsin hydrolysates were showed to have a higher degree of hydrolysis (24-33%), resulting in higher antioxidant and antibacterial activities. The white flower of trypsin hydrolysates showed a higher radical scavenging activity which could be attributed to its higher content of stress proteins (19%). However, trypsin hydrolysates from the purple flower showed higher ferric reducing power and bacterial growth inhibition. The performance of hydrolysates was better than ampicillin in inhibiting Acinetobacter baumanni and Staphylococcus aureus.
  5. Lai SL, Wong PF, Lim TK, Lin Q, Mustafa MR
    Proteomics, 2015 May;15(9):1608-21.
    PMID: 25594392 DOI: 10.1002/pmic.201400039
    Melanoma is a lethal form of skin cancer with rising global incidence. However, limited treatment options are available for advanced melanoma and this is further compounded by the development of resistance toward existing drugs. Panduratin A (PA), a cyclohexanyl chalcone found in Boesenbergia rotunda, was investigated for its cytotoxic potentials against human malignant melanoma A375 cells. Our initial findings revealed that mitochondrion is the primary acting site of PA on A375 cancer cells and the cytotoxic mechanisms of PA were further investigated using a temporal quantitative proteomics approach by iTRAQ 2D-LC-MS/MS. Comprehensive proteomics analysis identified 296 proteins that were significantly deregulated in PA-treated A375 cells and revealed the involvement of mitochondrial oxidative phosphorylation, secretory and ER stress pathway, and apoptosis. We further confirmed that the PA-induced apoptosis was mediated by prolonged ER stress at least in part via the PERK/eIF2α/ATF4/CHOP pathway. Pretreatment with cycloheximide, an ER stress inhibitor rescued PA-induced cell death, which was accompanied by the suppression of ER-stress-related HSPA5 and CHOP proteins. The present study provides comprehensive mechanistic insights into the cytotoxic mechanisms of PA.
  6. Lai SL, Wong PF, Lim TK, Lin Q, Mustafa MR
    Phytomedicine, 2015 Jan 15;22(1):203-12.
    PMID: 25636890 DOI: 10.1016/j.phymed.2014.11.016
    Panduratin A (PA), a cyclohexanyl chalcone from Boesenbergia rotunda (L.) Mansf. was shown to possess anti-angiogenic effects in our previous study. In the present study, the molecular targets and anti-angiogenic mechanisms of PA on human umbilical vein endothelial cells (HUVECs) were identified using an iTRAQ-based quantitative proteomics approach. A total of 263 proteins were found to be differentially regulated in response to treatment with PA. Ingenuity Pathway Analysis revealed that cellular growth and proliferation, protein synthesis, RNA post-transcriptional modification, cellular assembly and organization and cell-to-cell signaling and interaction were the most significantly deregulated molecular and cellular functions in PA-treated HUVECs. PA inhibited the expressions of ARPC2 and CTNND1 that are associated with the formation of actin cytoskeleton, focal adhesion and cellular protrusions. In addition, PA down-regulated CD63, GRB-2, ICAM-2 and STAB-1 that are implicated in adhesion, migration and tube formation of endothelial cells. The differential expressions of three targets, namely, ARPC2, CDK4, and GRB-2 were validated by western blot analyses. Furthermore, PA inhibited G1-S progression, and resulted in G0/G1 arrest in HUVECs. The blockage in cell cycle progression was accompanied with the suppression of mTOR signaling. Treatment of HUVECs with PA resulted in decreased phosphorylation of ribosomal S6 and 4EBP1 proteins, the two downstream effectors of mTOR signaling. We further showed that PA is able to inhibit mTOR signaling induced by VEGF, a potent inducer of angiogenesis. Taken together, by integrating quantitative proteomic approach, we identified protein targets in which PA mediates its anti-angiogenic effects. The present study thus provides mechanistic evidence to the previously reported multifaceted anti-angiogenic effects of PA. Our study further identified mTOR signaling as an important target of PA, and therefore highlights the potential of PA for therapeutic intervention against angiogenesis-related pathogenesis, particularly, metastatic malignancy.
  7. Fish-Low CY, Than LTL, Ling KH, Lin Q, Sekawi Z
    J Microbiol Immunol Infect, 2020 Feb;53(1):157-162.
    PMID: 31029530 DOI: 10.1016/j.jmii.2018.12.015
    BACKGROUND: Human leptospirosis, or commonly known as "rat urine disease" is a zoonotic disease that is caused by the bacteria called Leptospira sp. The incidence rate of leptospirosis has been under-reported due to its unspecific clinical symptoms and the limitations of current laboratory diagnostic methods. Leptospirosis can be effectively treated with antibiotics in the early stage, and it is a curable disease but the accuracy to diagnose the infection is rarely achieved.

    METHODS: The present pilot study investigated plasma protein profiles of leptospirosis patients and compared them against two control groups which consisted of dengue patients and healthy individuals. The plasma protein digests were analyzed using shotgun approach by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Protein abundances were estimated from the exponentially modified protein abundance index (emPAI) values. Plasma proteins in leptospirosis patients with at least two-fold differential expression compared to dengue and healthy control groups (p 

  8. Qi Y, Hamzah SH, Gu E, Wang H, Xi Y, Sun M, et al.
    Nutrients, 2021 Jul 28;13(8).
    PMID: 34444765 DOI: 10.3390/nu13082605
    School gardening activities (SGA) combined with physical activities (PA) may improve childhood dietary intake and prevent overweight and obesity. This study aims to evaluate the effect of SGA combined with PA on children's dietary intake and anthropometric outcomes. We searched studies containing randomized controlled trials up to January 2021 in Web of Science, PubMed, Cochrane Library, and the EBSCO database on this topic for children aged 7 to 12 years. Fourteen studies met the requirements for meta-analysis (n = 9187). We found that SGA has no obvious effect on improving children's BMI (WMD = -0.49; p = 0.085; I2 = 86.3%), BMI z-score (WMD = -0.12; p = 0.235; I2 = 63.0%), and WC (WMD = -0.98; p = 0.05; I2 = 72.9%). SGA can effectively improve children's FVs (WMD = 0.59, p = 0.003, I2 = 95.3%). SGA combined with PA can significantly increase children's FVs but cannot greatly improve weight status. Although more studies on this topic are needed to prove the effectiveness of this method, the results of our review show that both SGA and SGA combined with PA has a modest but positive impact of reducing BMI and WC outcomes but can significantly increase children's FVs.
  9. Ng YL, Olivos-García A, Lim TK, Noordin R, Lin Q, Othman N
    Am J Trop Med Hyg, 2018 12;99(6):1518-1529.
    PMID: 30298805 DOI: 10.4269/ajtmh.18-0415
    Entamoeba histolytica is a protozoan parasite that causes amebiasis and poses a significant health risk for populations in endemic areas. The molecular mechanisms involved in the pathogenesis and regulation of the parasite are not well characterized. We aimed to identify and quantify the differentially abundant membrane proteins by comparing the membrane proteins of virulent and avirulent variants of E. histolytica HM-1:IMSS, and to investigate the potential associations among the differentially abundant membrane proteins. We performed quantitative proteomics analysis using isobaric tags for relative and absolute quantitation labeling, in combination with two mass spectrometry instruments, that is, nano-liquid chromatography (nanoLC)-matrix-assisted laser desorption/ionization-mass spectrometry/mass spectrometry and nanoLC-electrospray ionization tandem mass spectrometry. Overall, 37 membrane proteins were found to be differentially abundant, whereby 19 and 18 membrane proteins of the virulent variant of E. histolytica increased and decreased in abundance, respectively. Proteins that were differentially abundant include Rho family GTPase, calreticulin, a 70-kDa heat shock protein, and hypothetical proteins. Analysis by Protein ANalysis THrough Evolutionary Relationships database revealed that the differentially abundant membrane proteins were mainly involved in catalytic activities (29.7%) and metabolic processes (32.4%). Differentially abundant membrane proteins that were found to be involved mainly in the catalytic activities and the metabolic processes were highlighted together with their putative roles in relation to the virulence. Further investigations should be performed to elucidate the roles of these proteins in E. histolytica pathogenesis.
  10. Thevendran R, Navien TN, Meng X, Wen K, Lin Q, Sarah S, et al.
    Anal Biochem, 2020 07 01;600:113742.
    PMID: 32315616 DOI: 10.1016/j.ab.2020.113742
    The performance of aptamers as versatile tools in numerous analytical applications is critically dependent on their high target binding specificity and selectivity. However, only the technical or methodological aspects of measuring aptamer-target binding affinities are focused, ignoring the equally important mathematical components that play pivotal roles in affinity measurements. In this study, we aim to provide a comprehensive review regarding the utilization of different mathematical models and equations, along with a detailed description of the computational steps involved in mathematically deriving the binding affinity of aptamers against their specific target molecules. Mathematical models ranging from one-site binding to multiple aptameric binding site-based models are explained in detail. Models applied in several different approaches of affinity measurements such as thermodynamics and kinetic analysis, including cooperativity and competitive-assay based mathematical models have been elaborately discussed. Mathematical models incorporating factors that could potentially affect affinity measurements are also further scrutinized.
  11. Qi Y, Rong S, Liao K, Huo J, Lin Q, Hamzah SH
    Int J Environ Res Public Health, 2022 Oct 28;19(21).
    PMID: 36360973 DOI: 10.3390/ijerph192114096
    Inadequate intake of fruits and vegetables (FV) and moderate-to-vigorous physical activity (MVPA) in children has become a global public health problem. Therefore, school-based gardening and cooking (SGC) and sports participation (SP) interventions may be effective in improving children's FV intake and MVPA. The aim of this study is to develop and evaluate the effectiveness of SGC and SP interventions on FV intake and MVPA among Chinese children. In this cluster randomized controlled trial study, 237 children in grades 4-5 from six public primary schools from Changsha, Hunan Province, China will be randomly assigned to: (1) a SGC and SP combined intervention group; (2) a SP intervention group; (3) a regular practice group. The intervention clusters will be implemented for a period of 6 months and follow up will be carried out after 12 months. The outcome will be collected using a combination of self-reported and objective measures. Primary outcomes will include children's FV intake and duration of MVPA per day, and secondary outcomes will included frequency and attitudes of FV intake and SP, in addition to other measures. Finally, a process evaluation will be used to analyze the facilitators and barriers to intervention implementation. Trial Registration: (Registration Number: ChiCTR2200064141).
  12. Meng Z, Han J, Lin Y, Zhao Y, Lin Q, Ma X, et al.
    Theor Appl Genet, 2020 Jan;133(1):187-199.
    PMID: 31587087 DOI: 10.1007/s00122-019-03450-w
    KEY MESSAGE: A novel tetraploid S. spontaneum with basic chromosome x = 10 was discovered, providing us insights in the origin and evolution in Saccharum species. Sugarcane (Saccharum spp., Poaceae) is a leading crop for sugar production providing 80% of the world's sugar. However, the genetic and genomic complexities of this crop such as its high polyploidy level and highly variable chromosome numbers have significantly hindered the studies in deciphering the genomic structure and evolution of sugarcane. Here, we developed the first set of oligonucleotide (oligo)-based probes based on the S. spontaneum genome (x = 8), which can be used to simultaneously distinguish each of the 64 chromosomes of octaploid S. spontaneum SES208 (2n = 8x = 64) through fluorescence in situ hybridization (FISH). By comparative FISH assay, we confirmed the chromosomal rearrangements of S. spontaneum (x = 8) and S. officinarum (2n = 8x = 80), the main contributors of modern sugarcane cultivars. In addition, we examined a S. spontaneum accession, Np-X, with 2n = 40 chromosomes, and we found that it was a tetraploid with the unusual basic chromosome number of x = 10. Assays at the cytological and DNA levels demonstrated its close relationship with S. spontaneum with basic chromosome number x = 8 (the most common accessions in S. spontaneum), confirming its S. spontaneum identity. Population genetic structure and phylogenetic relationship analyses between Np-X and 64 S. spontaneum accessions revealed that Np-X belongs to the ancient Pan-Malaysia group, indicating a close relationship to S. spontaneum with basic chromosome number of x = 8. This finding of a tetraploid S. spontaneum with basic chromosome number of x = 10 suggested a parallel evolution path of genomes and polyploid series in S. spontaneum with different basic chromosome numbers.
  13. Chan SY, Chan BQY, Liu Z, Parikh BH, Zhang K, Lin Q, et al.
    ACS Omega, 2017 Dec 31;2(12):8959-8968.
    PMID: 30023596 DOI: 10.1021/acsomega.7b01604
    Natural polysaccharide pectin has for the first time been grafted with polyhydroxybutyrate (PHB) via ring-opening polymerization of β-butyrolactone. This copolymer, pectin-polyhydroxybutyrate (pec-PHB), was blended with PHB in various proportions and electrospun to produce nanofibers that exhibited uniform and bead-free nanostructures, suggesting the miscibility of PHB and pec-PHB. These nanofiber blends exhibited reduced fiber diameters from 499 to 336-426 nm and water contact angles from 123.8 to 88.2° on incorporation of pec-PHB. They also displayed 39-335% enhancement of elongation at break relative to pristine PHB nanofibers. pec-PHB nanofibers were found to be noncytotoxic and biocompatible. Human retinal pigmented epithelium (ARPE-19) cells were seeded onto pristine PHB and pec-PHB nanofibers as scaffold and showed good proliferation. Higher proportions of pec-PHB (pec-PHB10 and pec-PHB20) yielded higher densities of cells with similar characteristics to normal RPE cells. We propose, therefore, that nanofibers of pec-PHB have significant potential as retinal tissue engineering scaffold materials.
  14. Zhang Z, Hu Y, Ji H, Lin Q, Li X, Sang S, et al.
    Food Chem, 2023 Jul 30;415:135736.
    PMID: 36863232 DOI: 10.1016/j.foodchem.2023.135736
    Core-shell biopolymer nanoparticles are assembled from a hydrophobic protein (zein) core and a hydrophilic polysaccharide (carboxymethyl dextrin) shell. The nanoparticles were shown to have good stability and the ability to protect quercetin from chemical degradation under long-term storage, pasteurization, and UV irradiation. Spectroscopy analysis shows that electrostatic, hydrogen bonding, and hydrophobic interactions are the main driving forces for the formation of composite nanoparticles. Quercetin coated with nanoparticles significantly enhanced its antioxidant and antibacterial activities and showed good stability and slow release in vitro during simulated gastrointestinal digestion. Furthermore, the encapsulation efficiency of carboxymethyl dextrin-coated zein nanoparticles (81.2%) for quercetin was significantly improved compared with that of zein nanoparticles alone (58.4%). These results indicate that carboxymethyl dextrin-coated zein nanoparticles can significantly improve the bioavailability of hydrophobic nutrient molecules such as quercetin and provide a valuable reference for their application in the field of biological delivery of energy drinks and food.
  15. Mai W, Ren Y, Tian X, Al-Mahdi AY, Peng R, An J, et al.
    J Med Virol, 2023 Apr;95(4):e28692.
    PMID: 36946502 DOI: 10.1002/jmv.28692
    The coronavirus disease 2019 (COVID-19) pandemic and related public health intervention measures have been reported to have resulted in the reduction of infections caused by influenza viruses and other common respiratory viruses. However, the influence may be varied in areas that have different ecological, economic, and social conditions. This study investigated the changing epidemiology of 8 common respiratory pathogens, including Influenza A (IFVA), Influenza B (IFVB), Respiratory syncytial virus (HRSV), rhinovirus (RV), Human metapneumovirus Adenovirus, Human bocavirus, and Mycoplasma pneumoniae, among hospitalized children during spring and early summer in 2019-2021 in two hospitals in Hainan Island, China, in the COVID-19 pandemic era. The results revealed a significant reduction in the prevalence of IFVA and IFVB in 2020 and 2021 than in 2019, whereas the prevalence of HRSV increased, and it became the dominant viral pathogen in 2021. RV was one of the leading pathogens in the 3 year period, where no significant difference was observed. Phylogenetic analysis revealed close relationships among the circulating respiratory viruses. Large scale studies are needed to study the changing epidemiology of seasonal respiratory viruses to inform responses to future respiratory virus pandemics.
  16. Grey AC, Lin Q, Low TY, Wu W, Haynes PA, Chung MCM, et al.
    Mol Cell Proteomics, 2023 Sep;22(9):100627.
    PMID: 37532177 DOI: 10.1016/j.mcpro.2023.100627
    As the first in-person Asia Oceania Human Proteomics Organization (AOHUPO) congress since 2018, the 11th AOHUPO congress was an opportune time for the research community to reconnect and to renew friendships after the long period of restricted travel due to the global pandemic. Moreover, this congress was a great opportunity for the many AO regional proteomics and mass spectrometry scientists to meet in Singapore to exchange ideas and to present their latest findings. Cohosted by the Singapore Society for Mass Spectrometry and the Malaysian Proteomics Society and held in conjunction with the seventh Asia Oceania Agricultural Proteomics Organization Congress and Singapore Society for Mass Spectrometry 2023, the meeting featured both human and agricultural proteomics. Over five hundred scientists from the AO region converged on the MAX Atria @ Singapore EXPO, Changi, Singapore from May 8 to 10 for the main congress. The diverse program was made up of 64 invited speakers and panellists for seven plenary lectures, 27 concurrent symposia, precongress and postcongress workshops, and 174 poster presentations. The AOHUPO society were able to celebrate not only their 20th anniversary but also the outstanding academic research from biological and agricultural proteomics and related 'omics fields being conducted across the Asia-Oceania region.
  17. Tang C, Yang M, Fang Y, Luo Y, Gao S, Xiao X, et al.
    Nat Plants, 2016 05 23;2(6):16073.
    PMID: 27255837 DOI: 10.1038/nplants.2016.73
    The Para rubber tree (Hevea brasiliensis) is an economically important tropical tree species that produces natural rubber, an essential industrial raw material. Here we present a high-quality genome assembly of this species (1.37 Gb, scaffold N50 = 1.28 Mb) that covers 93.8% of the genome (1.47 Gb) and harbours 43,792 predicted protein-coding genes. A striking expansion of the REF/SRPP (rubber elongation factor/small rubber particle protein) gene family and its divergence into several laticifer-specific isoforms seem crucial for rubber biosynthesis. The REF/SRPP family has isoforms with sizes similar to or larger than SRPP1 (204 amino acids) in 17 other plants examined, but no isoforms with similar sizes to REF1 (138 amino acids), the predominant molecular variant. A pivotal point in Hevea evolution was the emergence of REF1, which is located on the surface of large rubber particles that account for 93% of rubber in the latex (despite constituting only 6% of total rubber particles, large and small). The stringent control of ethylene synthesis under active ethylene signalling and response in laticifers resolves a longstanding mystery of ethylene stimulation in rubber production. Our study, which includes the re-sequencing of five other Hevea cultivars and extensive RNA-seq data, provides a valuable resource for functional genomics and tools for breeding elite Hevea cultivars.
  18. Cao Y, Chen L, Chen H, Cun Y, Dai X, Du H, et al.
    Natl Sci Rev, 2023 Apr;10(4):nwac287.
    PMID: 37089192 DOI: 10.1093/nsr/nwac287
  19. Klionsky DJ, Abdel-Aziz AK, Abdelfatah S, Abdellatif M, Abdoli A, Abel S, et al.
    Autophagy, 2021 Jan;17(1):1-382.
    PMID: 33634751 DOI: 10.1080/15548627.2020.1797280
    In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.
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