Pei-Jane Tsai

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.

Visible-light-induced bactericidal activity of a nitrogen-doped titanium photocatalyst against human pathogens.

ABSTRACT The antibacterial activity of photocatalytic titanium dioxide (TiO2) substrates is induced primarily by UV light irradiation. Recently, nitrogen- and carbon-doped TiO2 substrates were shown to exhibit photocatalytic activities under visible-light illumination. Their antibacterial activity, however, remains to be quantified. In this study, we demonstrated that nitrogen-doped TiO2 substrates have superior visible-light-induced bactericidal activity against Escherichia coli compared to pure TiO2 and carbon-doped TiO2 substrates. We also found that protein- and light-absorbing contaminants partially reduce the bactericidal activity of nitrogen-doped TiO2 substrates due to their light-shielding effects. In the pathogen-killing experiment, a significantly higher proportion of all tested pathogens, including Shigella flexneri, Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus, Streptococcus pyogenes, and Acinetobacter baumannii, were killed by visible-light-illuminated nitrogen-doped TiO2 substrates than by pure TiO2 substrates. These findings suggest that nitrogen-doped TiO2 has potential application in the development of alternative disinfectants for environmental and medical usages.

Functional Fe3O4/TiO2 Core/Shell Magnetic Nanoparticles as Photokilling Agents for Pathogenic Bacteria

A photokilling approach for pathogenic bacteria is demonstrated using a new type of magnetic nanoprobe as the photokilling agent. In addition to their magnetic property, the nanoprobes have other features including a photocatalytic property and the capacity to target bacteria. The nanoprobes comprise iron oxide/titania (Fe(3)O(4)@TiO(2)) core/shell magnetic nanoparticles. As dopamine molecules can self-assemble onto the surface of the titania substrate, dopamine is used as the linker to immobilize succinic anhydride onto the surfaces of the Fe(3)O(4)@TiO(2) nanoparticles. This is followed by the immobilization of IgG via amide bonding. We demonstrate that the IgG-Fe(3)O(4)@TiO(2) magnetic nanoparticles not only have the capacity to target several pathogenic bacteria, but they also can effectively inhibit the cell growth of the bacteria targeted by the nanoparticles under irradiation of a low-power UV lamp within a short period. Staphylococcus saprophyticus, Streptococcus pyogenes, and antibiotic-resistant bacterial strains, such as multiantibiotic-resistant S. pyogenes and methicillin-resistant Staphylococcus aureus (MRSA), are used to demonstrate the feasibility of this approach.

Functional gold nanoparticles as photothermal agents for selective-killing of pathogenic bacteria

AIMS Our aim was to demonstrate that functional gold nanoparticles can be used as photothermal agents for the selective killing of pathogenic bacteria. MATERIALS & METHODS Gold nanoparticles with polygonal shapes, capable of absorbing near infrared (NIR) light, were generated through a photochemical reaction. Vancomycin, which can bind with the terminal D-Ala-D-Ala moieties of the peptide units of pathogen cell walls, was immobilized on the surface of the gold nanoparticles. The vancomycin-bound gold nanoparticles were used as the photothermal agents for the inhibition of pathogenic bacteria growth, under irradiation of NIR light (808 nm). RESULTS & DISCUSSION We have demonstrated that vancomycin-bound gold nanoparticles are capable of selective-binding onto the cell walls of pathogenic bacteria. A large portion (>99%) of bacteria targeted by the gold nanoparticles was destroyed under illumination by NIR light within 5 min owing to suffering from heating. CONCLUSIONS This photothermal approach is effective for the inhibition of pathogenic bacteria cell growth, including Gram-positive bacteria, Gram-negative bacteria and antibiotic-resistant bacteria.

Affinity Capture of Uropathogenic Escherichia coli Using Pigeon Ovalbumin-Bound Fe3O4@Al2O3 Magnetic Nanoparticles

Escherichia coli and Staphylococcus saprophyticus are the most common causes of urinary tract infections, with 80% of these infections caused by uropathogenic E. coli. Because the P fimbriae of E. coli have specificity toward Gal(alpha1-4)Gal beta units, pigeon ovalbumin (POA), whose structure contains terminal Gal(alpha1-4)Gal beta moieties, was used as a probe for interaction with P fimbriated E. coli. The functional affinity probes for these bacteria by immobilizing POA--a phosphoprotein--onto the surface of magnetic iron oxide nanoparticles (NPs) coated with alumina (Fe3O4@Al2O3), using the phosphate units of POA as linking groups for the formation of phosphate-alumina complexes. The immobilization process occurred within 30 s when performing the reaction under microwave heating. The magnetic POA-Fe3O4@Al2O3 NPs generated using this facile approach exhibited specificity toward P fimbriated E. coli. The bacteria targeted by the affinity probes were characterized by matrix-assisted laser desorption/ionization mass spectrometry. The detection limit toward uropathogenic bacteria when using this approach was approximately 9.60 x 10(4) cfu/mL (0.5 mL).

The effects of the bacterial interaction with visible-light responsive titania photocatalyst on the bactericidal performance

Bactericidal activity of traditional titanium dioxide (TiO2) photocatalyst is effective only upon irradiation by ultraviolet light, which restricts the potential applications of TiO2 for use in our living environments. Recently carbon-containing TiO2 was found to be photoactive at visible-light illumination that affords the potential to overcome this problem; although, the bactericidal activity of these photocatalysts is relatively lower than conventional disinfectants. Evidenced from scanning electron microscopy and confocal Raman spectral mapping analysis, we found the interaction with bacteria was significantly enhanced in these anatase/rutile mixed-phase carbon-containing TiO2. Bacteria-killing experiments indicate that a significantly higher proportion of all tested pathogens including Staphylococcus aureus, Shigella flexneri and Acinetobacter baumannii, were eliminated by the new nanoparticle with higher bacterial interaction property. These findings suggest the created materials with high bacterial interaction ability might be a useful strategy to improve the antimicrobial activity of visible-light-activated TiO2.

Toll-like receptor 2 deficiency improves insulin sensitivity and hepatic insulin signalling in the mouse

Aims/hypothesisSubstantial evidence suggests a link between elevated inflammation and development of insulin resistance. Toll-like receptor 2 (TLR2) recognises a large number of lipid-containing molecules and transduces inflammatory signalling in a variety of cell types, including insulin-responsive cells. Considering the contribution of the fatty acid composition in TLR2-depedent signalling, we hypothesised that the inflammatory signals transduced by TLR2 contribute to insulin resistance.MethodsMice deficient in TLR2 were used to investigate the in vivo roles of TLR2 in initiating and maintaining inflammation-associated insulin resistance and energy homeostasis.ResultsWe first recapitulated the observation with elevated expression of TLR2 and inflammatory cytokines in white adipose tissue and liver of ob/ob mice. Aged or high-fat-fed TLR2-deficient mice were protected from obesity and adipocyte hypertrophy compared with wild-type mice. Moreover, mice lacking TLR2 exhibited improved glucose tolerance and insulin sensitivity regardless of feeding them regular chow or a high-fat diet. This is accompanied by reductions in expression of inflammatory cytokines and activation of extracellular signal-regulated kinase (ERK) in a liver-specific manner. The attenuated hepatic inflammatory cytokine expression and related signalling are correlated with increased insulin action specifically in the liver in TLR2-deficient mice, reflected by increased insulin-stimulated protein kinase B (Akt) phosphorylation and IRS1 tyrosine phosphorylation and increased insulin-suppressed hepatocyte glucose production.Conclusions/interpretationThe absence of TLR2 attenuates local inflammatory cytokine expression and related signalling and increases insulin action specifically in the liver. Thus, our work has identified TLR2 as a key mediator of hepatic inflammation-related signalling and insulin resistance.

Functional Nanoparticle-Based Proteomic Strategies for Characterization of Pathogenic Bacteria

Although matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) can be employed to rapidly characterize pathogenic bacteria, bacterial cultures are generally required to obtain sufficient quantities of the bacterial cells prior to MALDI MS analysis. If this time-consuming step could be eliminated, the length of time required for identification of bacterial strains would be greatly reduced. In this paper, we propose an effective means of rapidly identifying bacteria--one that does not require bacterial culturing--using functional nanoparticle-based proteomic strategies that are characterized by extremely short analysis time. In this approach, we used titania-coated magnetic iron oxide nanoparticles (Fe(3)O(4)@TiO(2) NPs) as affinity probes to concentrate the target bacteria. The magnetic properties of the Fe(3)O(4)@TiO(2) NPs allow the conjugated target species to be rapidly isolated from the sample solutions under a magnetic field. Taking advantage of the absorption of the magnetic Fe(3)O(4) NPs in the microwave region of the electromagnetic spectrum, we performed the tryptic digestion of the captured bacteria under microwave heating for only 1-1.5 min prior to MALDI MS analysis. We identified the resulting biomarker ions by combining their MS/MS analysis results with protein database searches. Using this technique, we identified potential biomarker ions representing five gram-negative bacteria: Escherichia coli O157:H7, uropathogenic E. coli, Shigella sonnei, Pseudomonas aeruginosa, and Klebsiella pneumoniae. Finally, we demonstrated the practical feasibility of using this approach to rapidly characterize bacteria in clinical samples.

Invasive Group A Streptococcal Disease in Taiwan Is Not Associated with the Presence of Streptococcal Pyrogenic Exotoxin Genes

We reviewed the clinical features of 44 patients with invasive group A streptococcal (GAS) disease who were treated at two teaching hospitals in southern Taiwan from 1991 to 1994. Genes encoding streptococcal pyrogenic exotoxin types A (speA), B (speB), C (speC), and F (speF) and serotypes of M1, M6, and M12 were determined by polymerase chain reaction to target specific sequences in the 44 isolates recovered from these patients and in 28 isolates recovered from upper respiratory sites in 28 additional patients during the study period. The protease activity of these isolates was tested by using the casein plate method. Of the 44 patients with invasive diseases, 25 (57%) had no obvious underlying diseases, and 14 (32%) had preexisting neoplastic diseases or had previously used steroids. Twenty-five patients (57%) presented with cellulitis or necrotizing fasciitis, 24 (55%) had bacteremia, and eight (18%) had streptococcal toxic shock syndrome (STSS). Eight patients (18%) died of invasive GAS disease; seven had STSS, and seven had underlying diseases. All eight patients died within 48 hours after hospitalization. The presence of speA, speC, or speF was not implicated in any particular clinical syndrome in patients with invasive GAS disease. High-level protease activity and the M1 serotype of the isolates were significantly associated with the clinical signs of STSS and with mortality. M1 serotype and protease activity, as well as host immune status, might play significant roles in the pathogenesis of invasive GAS disease in Taiwan.

Antimicrobial Susceptibilities and Molecular Epidemiology of Clinical Isolates of Clostridium difficile in Taiwan

ABSTRACT The antimicrobial susceptibility and virulence factors of Clostridium difficile clinical isolates in Taiwan have not previously been reported. One hundred and thirteen isolates were collected from two major teaching hospitals in Taiwan from 2001 to 2009. Molecular typing was performed by an automated repetitive extragenic palindromic sequence-based PCR (rep-PCR) method (DiversiLab; Bacterial Barcodes, Inc., Athens, GA) and PCR ribotyping. Detection of tcdA, tcdB, cdtA, and cdtB genes was performed using a multiplex PCR assay, and gyrA and gyrB genes of moxifloxacin-nonsusceptible isolates were sequenced. All isolates were susceptible to vancomycin and metronidazole. Ninety-five (84%) isolates were susceptible to moxifloxacin, and the MIC90 for nemonoxacin was 4 μg/ml. Tigecycline showed favorable antibacterial activity (MIC90 of 0.06 μg/ml). Thirteen rep-PCR types were identified as a predominant rep-PCR type (type A; non-North American pulsed-field gel electrophoresis type 1 [NAP1], -NAP7, or -NAP8) accounting for 52.2% (59 isolates). Nine of 18 moxifloxacin-nonsusceptible isolates belonged to the rep-PCR type A. The rep-PCR type A and C isolates were distinct from NAP1 (ribotype 027) and NAP8 (ribotype 078) as determined by PCR ribotyping. Seventy-four (65%) isolates harbored tcdA and tcdB, and 15 (13%) harbored cdtAB encoding binary toxin. Eleven isolates had a gene deletion in tcdC, including a 39-bp deletion (9 isolates) and an 18-bp deletion (2). In conclusion, dissemination of a predominant C. difficile clone in southern and northern Taiwan was noted. However, no NAP1 (ribotype 027) isolate could be discovered in this study.