Touchable 3D hierarchically structured polyaniline nanoweb for capture and detection of pathogenic bacteria.

A bacteria-capturing platform is a critical function of accurate, quantitative, and sensitive identification of bacterial pathogens for potential usage in the detection of foodborne diseases. Despite the development of various nanostructures and their surface chemical modification strategies, relative to the principal physical contact propagation of bacterial infections, mechanically robust and nanostructured platforms that are available to capture bacteria remain a significant problem. Here, a three-dimensional (3D) hierarchically structured polyaniline nanoweb film is developed for the efficient capture of bacterial pathogens by hand-touching. This unique nanostructure ensures sufficient mechanical resistance when exposed to compression and shear forces and facilitates the 3D interfacial interactions between bacterial extracellular organelles and polyaniline surfaces. The bacterial pathogens (Escherichia coli O157:H7, Salmonella enteritidis, and Staphylococcus aureus) are efficiently captured through finger-touching, as verified by the polymerase chain reaction (PCR) analysis. Moreover, the real-time PCR results of finger-touched cells on a 3D nanoweb film show a highly sensitive detection of bacteria, which is similar to those of the real-time PCR using cultured cells without the capturing step without any interfering of fluorescence signal and structural deformation during thermal cycling.

Diversity, Pathogenicity, and Fungicide Sensitivity of Colletotrichum Species Associated with Apple Anthracnose in South Korea.

Apple fruits with anthracnose symptoms were collected from commercial apple orchards in different regions of the Republic of Korea, and isolations were made on potato dextrose agar to isolate the causal agents. The fungal isolates were identified based on their morphological characteristics, growth rates, and multigene sequences. Nine isolates were identified via phylogenetic analysis: three Colletotrichum fructicola, two C. fioriniae, one C. gloeosporioides sensu stricto (s.s.), two C. nymphaeae, and one C. siamense isolates. The pathogenicity of the Colletotrichum isolates was tested using detached apple fruits under laboratory conditions. This study also reidentified six Colletotrichum isolates responsible for apple anthracnose, which were deposited in the Korean Agricultural Culture Collection. Among the six isolates, three were identified as C. siamense (deposited as C. gloeosporioides s.s.), and three were C. nymphaeae (deposited as C. acutatum s.s.). All the Colletotrichum species identified in this study were highly sensitive to tebuconazole in terms of inhibition of mycelial growth (EC50 value of 0.12 to 2.1 µg/ml).

Development of zinc oxide-based sub-micro pillar arrays for on-site capture and DNA detection of foodborne pathogen.

Prevention and early detection of bacterial infection caused by foodborne pathogens are the most important task to human society. Although currently available diagnostic technologies have been developed and designed for detection of specific pathogens, suitable capturing tools for the pathogens are rarely studied. In this paper, a new methodology is developed and proposed to realize effective capturing through touchable flexible zinc oxide-based sub-micro pillar arrays through genetic analysis. Zinc oxide coated pillar arrays have a high surface area, flexible, and adheres strongly to bacteria. Therefore, it contributes to enhance the bacterial capturability. An in-depth analysis on the sub-sequential capturing process at the bacterial cell-pillar interface is presented. By carefully observing the structural changes and performing numerical analysis under different reaction times, the results are presented. The resulting zinc oxide coated pillar arrays exhibited comprehensive capturability. These pillars were able to detect pathogenic bacteria due to a combination of complex structures, depletion force, and high surface electrostatics. The developed sub-micro pillars successfully captured and detected infectious foodborne bacteria of Escherichia coli in the range of 106-101 CFU/mL.

Multifunctional hand-held sensor using electronic components embedded in smartphones for quick PCR screening.

We focused on the development of a hand-held pathogen-detection device using smartphone-embedded electronic elements combined with functionalized magnetic particles (MPs) and sepharose. To perform affinity chromatography for evaluating DNA amplicons, avidin-conjugated MPs and succinimide-linked sepharose were used with biotin-primers. To mimic the centrifugal-based affinity ligand chromatography, a smartphone-mountable low-power fan was plugged into the charging port of a smartphone. The charging port stably emitted electric current at 3.0 V, and the fan blades were modified for use as a portable rotor. Based on the binding variation of MPs with DNA amplicons, the position of MPs in sepharose changed significantly during centrifugation. The change in distance was optically analyzed using the illumination sensor of the smartphone with respect to the altered transmittance due to the MPs. Amplified genes from Escherichia. coli O157:H7 samples ranging from 1.0 × 101 to 1.0 × 106 colony-forming units could be rapidly and immediately detected by the naked eye using a simple smartphone-based optical device. The results indicated that this novel biosensing technique is suitable for use as a point-of-care testing device in both industrial and clinical fields.

Naked eye detection of an amplified gene using metal particle-based DNA transport within functionalized porous interfaces.

The current study focuses on developing a system for visually detecting an amplified bacterial (Escherichia coli O157:H7) gene using a heavy metal particle (MP) and functionalized porous sepharose gel. To functionalize DNA-specificity to the MP, an avidin-modified MP was employed in combination with a biotin-conjugated primer. The porous sepharose matrix was functionalized with an amine-reactive group, such as N-hydroxysuccinimide (NHS), to achieve separation upon binding of the amplified gene. The pristine avidin-MPs strongly react with NHS-sepharose via imide bonds owing to the exposure of the amine group on the avidin-MP surface. Conversely, together with the amplified gene, the avidin-MPs are relatively less interactive toward the sepharose gel by steric hindrance of the amplified gene toward the imide bond between NHS and the amine groups. Owing to the higher molecular mass of the MP, those metal particles complexed with the amplified gene pass through the sepharose matrix when centrifugal force is applied. The MPs that are thus separated can be easily visualized by the naked eye owing to their inherent reddish-brown color. A polymerase chain reaction (PCR) product of E. coli O157:H7, present in concentrations ranging from 1.0 × 101 to 1.0 × 106 colony forming units (CFU), in actual food sample was evaluated with high sensitivity and reproducibility. We expect that the MP-based sensing system, which allows for visual detection of PCR-amplified genes, can be clinically used as a point-of-care testing device.

Combined Fluid Shear Stress and Melatonin Enhances the ERK/Akt/mTOR Signal in Cilia-Less MC3T3-E1 Preosteoblast Cells.

We investigated whether combined fluid shear stress (FSS) and melatonin stimulated signal transduction in cilia-less MC3T3-E1 preosteoblast cells. MC3T3-E1 cells were treated with chloral hydrate or nocodazole, and mechanotransduction sensor primary cilia were removed. p-extracellular signal⁻regulated kinase (ERK) and p-Akt with/without melatonin increased with nocodazole treatment and decreased with chloral hydrate treatment, whereas p-ERK and p-Akt in FSS with/without melatonin increased in cilia-less groups compared to cilia groups. Furthermore, p-mammalian target of rapamycin (mTOR) with FSS-plus melatonin increased in cilia-less groups compared to only melatonin treatments in cilia groups. Expressions of Bcl-2, Cu/Zn-superoxide dismutase (SOD), and catalase proteins were higher in FSS with/without melatonin with cilia-less groups than only melatonin treatments in cilia groups. Bax protein expression was high in FSS-plus melatonin with chloral hydrate treatment. In chloral hydrate treatment with/without FSS, expressions of Cu/Zn-SOD, Mn-SOD, and catalase proteins were high compared to only-melatonin treatments. In nocodazole treatment, Mn-SOD protein expression without FSS was high, and catalase protein level with FSS was low, compared to only melatonin treatments. These data show that the combination with FSS and melatonin enhances ERK/Akt/mTOR signal in cilia-less MC3T3-E1, and the enhanced signaling in cilia-less MC3T3-E1 osteoblast cells may activate the anabolic effect for the preservation of cell structure and function.

Ultrasonic fabrication of flexible antibacterial ZnO nanopillar array film.

Antibacterial activity is essential and highly demanded in worldwide to prevent potential bacterial infection. Here in this work, we report a new approch for the fabrication of flexible zinc oxide nanopillar arrays (ZG-NPA) film with an efficient antibacterial activity. A flexible NPA film served as a substrate for the rapid formation of ZnO by using ultrasound-assisted method. The enhancement of antibacterial activity were induced by cellular damages because of nano topological effects and electrostatic interaction between bacteria and ZG-NPA. Owing to the benefits of combination with flexibility, high surface areas from nano-features and excellent antibacterial efficiency (>80%) of ZG-NPA, the film can show great potential for use as novel biomaterials for preventing bacterial infections.

A film-based integrated chip for gene amplification and electrochemical detection of pathogens causing foodborne illnesses.

Given the increased interest in public hygiene due to outbreaks of food poisoning, increased emphasis has been placed on developing novel monitoring systems for point-of-care testing (POCT) to evaluate pathogens causing foodborne illnesses. Here, we demonstrate a pathogen evaluation system utilizing simple film-based microfluidics, featuring simultaneous gene amplification, solution mixing, and electrochemical detection. To minimize and integrate the various functionalities into a single chip, patterned polyimide and polyester films were mainly used on a polycarbonate housing chip, allowing simple fabrication and alignment, in contrast to conventional polymerase chain reaction, which requires a complex biosensing system at a bench-top scale. The individual integrated sensing chip could be manually fabricated in 10 min. Using the developed film-based integrated biosensing chip, the genes from the pathogens causing foodborne illnesses were simultaneously amplified based on multiple designed microfluidic chambers and Hoechst 33258, which intercalates into double-stranded DNA, to generate the electrochemical signal. The target pathogen gene was accurately analyzed by square wave voltammetry (SWV) within the 25 s, while the gel electrophoresis required about 30 min. Based on the developed integrated biosensing chip, the 1.0 × 101 and 1.0 × 102 colony-forming unit (CFU) of Staphylococcus aureus and Escherichia coli were sensitively detected with high reproducibility in the 25 s. On the basis of the significant features of the film-based molecular analysis platform, we expect that the developed sensor could be applied to the screening of various pathogens as a POCT device.

Surface-Modified Mesh Filter for Direct Nucleic Acid Extraction and its Application to Gene Expression Analysis.

Rapid and convenient isolation of nucleic acids (NAs) from cell lysate plays a key role for onsite gene expression analysis. Here, this study achieves one-step and efficient capture of NA directly from cell lysate by developing a cationic surface-modified mesh filter (SMF). By depositing cationic polymer via vapor-phase deposition process, strong charge interaction is introduced on the surface of the SMF to capture the negatively charged NAs. The NA capturing capability of SMF is confirmed by X-ray photoelectron spectroscopy, fluorescent microscopy, and zeta potential measurement. In addition, the genomic DNAs of Escherichia Coli O157:H7 can be extracted by the SMF from artificially infected food, and fluorescent signal is observed on the surface of SMF after amplification of target gene. The proposed SMF is able to provide a more simplified, convenient, and fast extraction method and can be applied to the fields of food safety testing, clinical diagnosis, or environmental pollutant monitoring.

Maturation of Adipocytes is Suppressed by Fluid Shear Stress.

Preadipocytes are mechano-responsive cells and their differentiation to adipocytes may be regulated by various types of physical stimulation. Understanding the mechanism of differentiation, which increases the number of adipocytes and lipid accumulation is important in the study of obesity-related diseases. In this study, we investigated the effects of physical stimulation at different stages of adipogenic differentiation using physiological levels of fluid shear stress. Preadipocytes were treated with dexamethasone, 3-isobutyl-1-methylxanthine and insulin for 3 days (induction period) and incubated for additional 6 days for maturation. Fluid shear stress of 1 Pa at 1 Hz was applied for 1 h at different stages of differentiation. Fluid shear stress applied at the maturation period significantly reduced the expressions of C/enhancer binding protein (EBP)α and peroxisome proliferator-activated receptor (PPAR)γ2 leading to reduced lipid accumulation. Fluid shear stress applied at the early or late stages of the induction period only decreased peroxisome proliferator-activated receptor γ2 expression without any significant changes in lipid accumulation. Stimulation at multiple days during the induction period did not result in changes in lipid accumulation compared to stimulation at a single day. These results suggest that lipid droplet accumulation is effectively decreased by fluid shear stress applied during the cell maturation period. Understanding the cellular response to physical stimulation throughout the entire adipocyte differentiation period may be important in controlling adipogenesis by physical stimulation.

Flexible and Disposable Sensing Platforms Based on Newspaper.

The flexible sensing platform is a key component for the development of smart portable devices targeting healthcare, environmental monitoring, point-of-care diagnostics, and personal electronics. Herein, we demonstrate a simple, scalable, and cost-effective strategy for fabrication of a sensing electrode based on a waste newspaper with conformal coating of parylene C (P-paper). Thin polymeric layers over cellulose fibers allow the P-paper to possess improved mechanical and chemical stability, which results in high-performance flexible sensing platforms for the detection of pathogenic E. coli O157:H7 based on DNA hybridization. Moreover, P-paper electrodes have the potential to serve as disposable, flexible sensing platforms for point-of-care testing biosensors.

Mutational inactivation of OprD in carbapenem-resistant Pseudomonas aeruginosa isolates from Korean hospitals.

This study investigated the mechanisms underlying the carbapenem resistance of bloodstream isolates of Pseudomonas aeruginosa obtained from two Korean hospitals. Of the 79 P. aeruginosa isolates, 22 and 21 were resistant to imipenem and meropenem, respectively. The 22 imipenem-resistant P. aeruginosa isolates were classified into 7 sequence types (STs) and 13 pulsotypes. Twelve imipenem-resistant isolates from one hospital were found to belong to the international clone ST111. Two imipenem-resistant P. aeruginosa ST235 isolates carried the bla IMP-6 gene, but the remaining 20 isolates did not produce carbapenemases. Mutations in the oprD gene and a related decrease in gene expression were found in 21 and 5 isolates, respectively. However, all imipenemresistant P. aeruginosa isolates showed no significant expression of OprD in the outer membrane as compared with that of carbapenem-susceptible PAO1 strain. Overexpression of genes associated with efflux pumps, including mexB, mexD, mexF, and mexY, was not found in any imipenem-resistant isolate. One imipenem-resistant P. aeruginosa isolate overexpressed the ampC gene. Our results show that the low permeability of drugs due to the mutational inactivation of OprD is primarily responsible for carbapenem resistance in bloodstream isolates of P. aeruginosa from Korean hospitals.

Effects of neuropeptides and mechanical loading on bone cell resorption in vitro.

Neuropeptides such as vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP) are present in nerve fibers of bone tissues and have been suggested to potentially regulate bone remodeling. Oscillatory fluid flow (OFF)-induced shear stress is a potent signal in mechanotransduction that is capable of regulating both anabolic and catabolic bone remodeling. However, the interaction between neuropeptides and mechanical induction in bone remodeling is poorly understood. In this study, we attempted to quantify the effects of combined neuropeptides and mechanical stimuli on mRNA and protein expression related to bone resorption. Neuropeptides (VIP or CGRP) and/or OFF-induced shear stress were applied to MC3T3-E1 pre-osteoblastic cells and changes in receptor activator of nuclear factor kappa B (NF-κB) ligand (RANKL) and osteoprotegerin (OPG) mRNA and protein levels were quantified. Neuropeptides and OFF-induced shear stress similarly decreased RANKL and increased OPG levels compared to control. Changes were not further enhanced with combined neuropeptides and OFF-induced shear stress. These results suggest that neuropeptides CGRP and VIP have an important role in suppressing bone resorptive activities through RANKL/OPG pathway, similar to mechanical loading.

Photoluminescent green carbon nanodots from food-waste-derived sources: large-scale synthesis, properties, and biomedical applications.

We have developed a simple approach for the large-scale synthesis of water-soluble green carbon nanodots (G-dots) from many kinds of large food waste-derived sources. About 120 g of G-dots per 100 kg of food waste can be synthesized using our simple and environmentally friendly synthesis approach. The G-dots exhibit a high degree of solubility in water because of the abundant oxygen-containing functional groups around their surface. The narrow band of photoluminescence emission (400-470 nm) confirms that the size of the G-dots (∼4 nm) is small because of a similar quantum effects and emission traps on the surfaces. The G-dots have excellent photostability; their photoluminescence intensity decreases slowly (∼8%) under continuous excitation with a Xe lamp for 10 days. We carried out cell viability assay to assess the effect of cytotoxicity by introducing G-dots in cells such as Chinese hamster ovary cells (CHO-K1), mouse muscle cells (C2C12), and African green monkey kidney cells (COS-7), up to a concentration of 2 mg mL(-1) for 24 h. Due to their high photostability and low cytotoxicity, these G-dots are excellent probes for in vitro bioimaging. Moreover, the byproducts (not including G-dots) of G-dot synthesis from large food-waste derived sources promoted the growth and development of seedlings germinated on 3DW-supplemented gauze. Because of the combined advantages of green synthesis, high aqueous stability, high photostability, and low cytotoxicity, the G-dots show considerable promise in various areas, including biomedical imaging, solution state optoelectronics, and plant seed germination and/or growth.

Altered APP Carboxyl-Terminal Processing Under Ferrous Iron Treatment in PC12 Cells.

Amyloid-ß peptide (Aß), generated by proteolytic cleavage of the amyloid precursor protein (APP), plays a pivotal role in the pathogenesis of Alzheimer's disease (AD). The key step in the generation of Aß is cleavage of APP by beta-site APP-cleaving enzyme 1 (BACE1). Levels of BACE1 are increased in vulnerable regions of the AD brain, but the underlying mechanism is unknown. In the present study, we reported the effects of ferrous ions at subtoxic concentrations on the mRNA levels of BACE1 and a-disintegrin-and-metalloproteinase 10 (ADAM10) in PC12 cells and the cell responses to ferrous ions. The cell survival in PC12 cells significantly decreased with 0 to 0.3 mM FeCl2, with 0.6 mM FeCl2 treatment resulting in significant reductions by about 75%. 4,6-diamidino-2-phenylindole (DAPI) staining showed that the nuclei appeared fragmented in 0.2 and 0.3 mM FeCl2. APP-α-carboxyl terminal fragment (APP-α-CTF) associations with ADAM10 and APP-ß-CTF with BACE1 were increased. Levels of ADAM10 and BACE1 mRNA increased in response to the concentrations of 0.25 mM, respectively. In addition, p-ERK and p-Bad (S112, S155) expressions were increased, suggesting that APP-CTF formation is related to ADAM10/BACE1 expression. Levels of Bcl-2 protein were increased, but significant changes were not observed in the expression of Bax. These data suggest that ion-induced enhanced expression of AMDA10/BACE1 could be one of the causes for APP-α/ß-CTF activation.

Fluid shear stress and melatonin in combination activate anabolic proteins in MC3T3-E1 osteoblast cells.

In this study, we investigated whether fluid shear stress and melatonin in combination stimulate the anabolic proteins through the phosphorylation of extracellular signal-regulated kinase (p-ERK) in MC3T3-E1 osteoblast cells. First, we researched why fluid shear stress and melatonin in combination influence cell survival. Fluid shear stress (1 hr) and melatonin (1 mM) in combination reduced autophagic marker LC3-II compared with fluid shear stress (1 hr) and/or melatonin (0.1 mM). Under the same conditions for fluid shear stress, markers of cell survival signaling pathway p-ERK, phosphorylation of serine-threonine protein kinase (p-Akt), phosphorylation of mammalian target of rapamycin (p-mTOR), and p85-S6K were investigated. p-Akt, p-mTOR (Ser 2481) expressions increased with the addition of 1 mM melatonin prior to 0.1 mM melatonin treatment. However, p-S6K expression did not change significantly. Next, mitochondria activity including Bcl-2, Bax, catalase, and Mn-superoxide dismutase (Mn-SOD) were studied. Expressions of Bcl-2, Bax, and catalase proteins were low under fluid shear stress plus 1 mM melatonin compared with only fluid shear stress alone, whereas Mn-SOD expression was high compared with conditions of no fluid shear stress. Finally, the anabolic proteins of bone, osteoprotegerin, type I collagen (collagen I), and bone sialoprotein II (BSP II) were checked. These proteins increased with combined fluid shear stress (1, 4 hr) and melatonin (0.1, 1 mM). Together, these results suggest that fluid shear stress and melatonin in combination may increase the expression of anabolic proteins through the p-ERK in MC3T3-E1 osteoblast cells. Therefore, fluid shear stress in combination with melatonin may promote the anabolic response of osteoblasts.

A high-precision angular control system for HIFU calibration.

A design of high-precision angular position control system for calibrating high intensity focused ultrasound (HIFU) is presented with alignment procedures. Two independent angular controls are achieved by combining a worm gear and a belt gear system. The proposed system verifies alignment by comparing simulation data and experimental data with three different transducers and two different types of hydrophones. The performance of the proposed system is compared to that of a commercial system. The results indicate that the proposed system provides high precision angular alignment (e.g., <0.01radians) with robust reproducibility regardless of the hydrophone type.

Carbenoxolone prevents the development of fatty liver in C57BL/6-Lep ob/ob mice via the inhibition of sterol regulatory element binding protein-1c activity and apoptosis.

Carbenoxolone is the 3-hemisuccinate of glycyrrhetinic acid, the active principal of licorice (Glycyrrhiza glabra). It was reported that carbenoxolone improved glucose tolerance with increased insulin sensitivity in mice with high fat diet-induced obesity. In the present study, we elucidated the protective effect of carbenoxolone in fatty liver animal models of C57BL/6-Lep(ob/ob) mice through inhibition of hepatic lipogenesis and apoptosis. In addition, the potential mechanisms by which carbenoxolone could exert such protection were elucidated. Carbenoxolone was daily administrated by gavage for 28 days in C57BL/6 and C57BL/6-Lep(ob/ob) mice. Carbenoxolone prevented the plasma triglyceride and free fatty acid accumulation associated with the reduction of the expression of sterol regulatory element binding protein-1c, liver X receptor, fatty acid synthase and acethyl-CoA carboxylase in the livers of C57BL/6-Lep(ob/ob) mice. Carbenoxolone also prevented hepatic injury through anti-apoptotic action in the livers of C57BL/6-Lep(ob/ob) mice, accompanied by increased Bcl-2 expression and suppressed Bax and cytochrome c expression. As a mechanism, increased inflammatory cytokine expressions were inhibited by carbenoxolone in the fatty livers of C57BL/6-Lep(ob/ob) mice. Furthermore, carbenoxolone inhibited free fatty acid (oleate/palmitate) induced reactive oxygen species formation and reversed free fatty acid induced mitochondrial membrane depolarization in HepG2 cells. Carbenoxolone prevents the development of fatty liver by inhibiting sterol regulatory element binding protein-1c expression and activity with an anti-apoptotic mechanism via the inhibition of inflammatory cytokine and reactive oxygen species formation in the livers of C57BL/6-Lep(ob/ob) mice. It is suggested that carbenoxolone prevents the development and progression of fatty liver disease in patients with insulin resistance.

Melatonin-induced autophagy is associated with degradation of MyoD protein in C2C12 myoblast cells.

MyoD is a muscle-specific transcriptional factor that acts as a master switch for skeletal muscle differentiation. This protein regulates myoblast proliferation and myogenic differentiation and is also a short-lived regulatory protein that is degraded by the ubiquitin system. However, the lysosomal pathway of MyoD protein degradation remains unknown. In this study, we sought to determine whether melatonin (1, 2mm)-induced autophagy causes the degradation of MyoD protein in C2C12 myoblast cells. Melatonin induced a significant increase in expression of the microtubule-associated protein 1 light chain 3 (LC3)-II and Beclin-1 proteins in a dose-dependent manner. Melatonin treatment also significantly increased p-ERK, Ras, and p-Akt expressions in a dose-dependent manner. However, Bax expression was high compared with the absence of melatonin treatment, and Bcl-2 expression was high in the 0.1-0.5mm melatonin treatments and low in the 1 and 2mm melatonin treatments. Under the same conditions, cytosolic MyoD protein was significantly decreased in a dose-dependent manner and completely eliminated by 36hr. This decrease in MyoD protein involved ubiquitin-mediated proteasomal activity with proteasome inhibitor MG132 or autophagy-dependent lysosomal degradation with lysosomal inhibitor bafilomycin A1 (Baf-A1). In the same condition, phosphorylation of the mammalian target of rapamycin, p-mTOR, and p-S6K expression with Baf-A1 or Baf-A1-plus melatonin treatment were significantly decreased compared with the levels after treatment with melatonin only. Together, these results suggest that melatonin (1, 2mm)-induced autophagy results in partial lysosomal degradation of MyoD protein in C2C12 myoblast cells.

Protective effects of arachidonic acid against palmitic acid-mediated lipotoxicity in HIT-T15 cells.

Saturated fatty acids have been considered major contributing factors in type 2 diabetes, whereas unsaturated fatty acids have beneficial effects for preventing the development of diabetes. However, the effects of polyunsaturated fatty acids in pancreatic ß cells have not been reported. Here, we examined the effects of arachidonic acid (AA) on palmitic acid (PA)-mediated lipotoxicity in clonal HIT-T15 pancreatic ß cells. AA prevented the PA-induced lipotoxicity as indicated by cell viability, DNA fragmentation and mitochondrial membrane potential, whereas eicosatetraynoic acid (ETYA), a non-metabolizable AA, had little effect on PA-induced lipotoxicity. In parallel with its protective effects against PA-induced lipotoxicity, AA restored impaired insulin expression and secretion induced by PA. AA but not ETYA increased intracellular triglyceride (TG) in the presence of PA compared with PA alone, and xanthohumol, a diacylglycerol acyltransferase (DGAT) inhibitor, reversed AA-induced protection from PA. Taken together, our results suggest that AA protects against PA-induced lipotoxicity in clonal HIT-T15 pancreatic ß cells, and the protective effects may be associated with TG accumulation, possibly through sequestration of lipotoxic PA into TG.