speG Is Required for Intracellular Replication of Salmonella in Various Human Cells and Affects Its Polyamine Metabolism and Global Transcriptomes.

The speG gene has been reported to regulate polyamine metabolism in Escherichia coli and Shigella, but its role in Salmonella remains unknown. Our preliminary studies have revealed that speG widely affects the transcriptomes of infected in vitro M and Caco-2 cells and that it is required for the intracellular replication of Salmonella enterica serovar Typhimurium (S. Typhimurium) in HeLa cells. In this study, we demonstrated that speG plays a time-dependent and cell type-independent role in the intracellular replication of S. Typhimurium. Moreover, high-performance liquid chromatography (HPLC) of four major polyamines demonstrated putrescine, spermine, and cadaverine as the leading polyamines in S. Typhimurium. The deletion of speG significantly increased the levels of the three polyamines in intracellular S. Typhimurium, suggesting the inhibitory effect of speG on the biosynthesis of these polyamines. The deletion of speG was associated with elevated levels of these polyamines in the attenuated intracellular replication of S. Typhimurium in host cells. This result was subsequently validated by the dose-dependent suppression of intracellular proliferation after the addition of the polyamines. Furthermore, our RNA transcriptome analysis of S. Typhimurium SL1344 and its speG mutant outside and inside Caco-2 cells revealed that speG regulates the genes associated with flagellar biosynthesis, fimbrial expression, and functions of types III and I secretion systems. speG also affects the expression of genes that have been rarely reported to correlate with polyamine metabolism in Salmonella, including those associated with the periplasmic nitrate reductase system, glucarate metabolism, the phosphotransferase system, cytochromes, and the succinate reductase complex in S. Typhimurium in the mid-log growth phase, as well as those in the ilv-leu and histidine biosynthesis operons of intracellular S. Typhimurium after invasion in Caco-2 cells. In the present study, we characterized the phenotypes and transcriptome effects of speG in S. Typhimurium and reviewed the relevant literature to facilitate a more comprehensive understanding of the potential role of speG in the polyamine metabolism and virulence regulation of Salmonella.

Evaluation of tyrosinase inhibitory and antioxidant activities of Angelica dahurica root extracts for four different probiotic bacteria fermentations.

Angelica dahurica root (ADR), which shows strong antioxidant activity, is used in Chinese medicine. This study evaluated the tyrosinase inhibitory and antioxidant activities of ADR extracts fermented by four different probiotic bacteria: Bifidobacterium bifidum, Bifidobacterium lactis, Lactobacillus acidophilus, and Lactobacillus brevis. The ADR was first extracted using distilled water, 70% ethanol, and ethyl acetate, and then fermented by probiotic bacteria. The physiological characteristics of these fermented extracts, namely the antityrosinase activity, antioxidant activity, phenolic composition, and phenolic content, were evaluated and compared with those of unfermented extracts. Results showed that the water extracts after fermentation by probiotic bacteria exhibited the most favorable physiological characteristics. Among the extracts fermented by these probiotic bacteria, L. acidophilus-fermented ADR extract showed the most favorable physiological characteristics. The optimal IC50 values for antityrosinase activity, DPPH radical scavenging activity, and reducing power for L. acidophilus-fermented ADR extract were 0.07 ± 0.03, 0.12 ± 0.01, and 0.68 ± 0.06 mg/mL, respectively. Furthermore, the physiological activities of fermented extracts were considerably higher than those of unfermented extracts. The tyrosinase inhibition and melanin content of B16F10 melanoma cells, and cytotoxicity effects of the fermented ADR extracts on B16F10 cells were also evaluated. We found that the L. acidophilus-fermented ADR extract at 1.5 mg/mL showed significant cellular antityrosinase activity with low melanin production in B16F10 cells and was noncytotoxic to B16F10 cells. Among all probiotic bacteria, water-extracted ADR fermented by L. acidophilus for 48 h was found to be the best skincare agent or antioxidant agent.

Role of yqiC in the Pathogenicity of Salmonella and Innate Immune Responses of Human Intestinal Epithelium.

The yqiC gene of Salmonella enterica serovar Typhimurium (S. Typhimurium) regulates bacterial growth at different temperatures and mice survival after infection. However, the role of yqiC in bacterial colonization and host immunity remains unknown. We infected human LS174T, Caco-2, HeLa, and THP-1 cells with S. Typhimurium wild-type SL1344, its yqiC mutant, and its complemented strain. Bacterial colonization and internalization in the four cell lines significantly reduced on yqiC depletion. Post-infection production of interleukin-8 and human ß-defensin-3 in LS174T cells significantly reduced because of yqiC deleted in S. Typhimurium. The phenotype of yqiC mutant exhibited few and short flagella, fimbriae on the cell surface, enhanced biofilm formation, upregulated type-1 fimbriae expression, and reduced bacterial motility. Type-1 fimbriae, flagella, SPI-1, and SPI-2 gene expression was quantified using real-time PCR. The data show that deletion of yqiC upregulated fimA and fimZ expression and downregulated flhD, fliZ, invA, and sseB expression. Furthermore, thin-layer chromatography and high-performance liquid chromatography revealed the absence of menaquinone in the yqiC mutant, thus validating the importance of yqiC in the bacterial electron transport chain. Therefore, YqiC can negatively regulate FimZ for type-1 fimbriae expression and manipulate the functions of its downstream virulence factors including flagella, SPI-1, and SPI-2 effectors.

Hexavalent chromium removal and bioelectricity generation by Ochrobactrum sp. YC211 under different oxygen conditions.

Bioremediation is an environmentally friendly method of reducing heavy metal concentration and toxicity. A chromium-reducing bacterial strain, isolated from the vicinity of an electroplate factory, was identified as Ochrobactrum sp. YC211. The efficiency and capacity per time of Ochrobactrum sp. YC211 for hexavalent chromium (Cr(VI)) removal under anaerobic conditions were superior to those under aerobic conditions. An acceptable removal efficiency (96.5 ± 0.6%) corresponding to 30.2 ± 0.8 mg-Cr (g-dry cell weight-h)(-1) was achieved by Ochrobactrum sp. YC211 at 300 mg L(-1) Cr(VI). A temperature of 30°C and pH 7 were the optimal parameters for Cr(VI) removal. By examining reactivated cells, permeabilized cells, and cell-free extract, we determined that Cr(VI) removal by Ochrobactrum sp. YC211 under anaerobic conditions mainly occurred in the soluble fraction of the cell and can be regarded as an enzymatic reaction. The results also indicated that an Ochrobactrum sp. YC211 microbial fuel cell (MFC) with an anaerobic anode was considerably superior to that with an aerobic anode in bioelectricity generation and Cr(VI) removal. The maximum power density and Cr(VI) removal efficiency of the MFC were 445 ± 3.2 mW m(-2) and 97.2 ± 0.3%, respectively. Additionally, the effects of coexisting ions (Cu(2+), Zn(2+), Ni(2+), SO4(2-), and Cl(-)) in the anolyte on the MFC performance and Cr(VI) removal were nonsignificant (P > 0.05). To our knowledge, this is the first report to compare Cr(VI) removal by different cells and MFC types under aerobic and anaerobic conditions.

Injectable adipose tissue combined with stem cells for soft-tissue augmentation: A pilot study for dental applications.

BACKGROUND/PURPOSE: Bone resorption and soft-tissue defects are the typical physiologic responses after tooth extraction. Various dental ridge augmentation techniques have been applied and lack of the soft tissue is the major factor causing the failure. We propose that the adipose-derived stem cell can be useful in soft-tissue augmentation in dental applications. The objective of this study was to optimize the operation procedures for the isolation of adipose stem cells and tissues. Accelerated clinical protocols for effective transplantation of adipose tissue with high amount of adipose stem cells shall be developed. MATERIALS AND METHODS: Operation parameters were designed and optimized for the extraction of adipose tissue-derived stromal vascular cells. The optimized accelerated procedure was washing the lipoaspirate samples one time. Collagenase was then added and samples were incubated in a water bath for 30 minutes at 37°C and centrifuged at 1200g for 3 minutes. A mouse animal model was applied to evaluate the soft-tissue-filling effects using the optimized procedure. RESULTS: The animal model tests demonstrated the filling and regeneration of the soft tissues with significant angiogenesis. CONCLUSION: This pilot study demonstrated the feasibility of soft-tissue augmentation applications.

Effect of colour LEDs on mycelia growth of Aspergillus ficuum and phytase production in photo-fermentations.

Aspergillus ficuum grown on plates and in liquid cultures were illuminated by a white fluorescent light and four different colour LED lights (white, blue, green and red) to evaluate the regulation of LED lights on fungal growth. Biomass conversion, pellet size and phytase activity were examined. In liquid culture, luminous intensity was highly correlated with the rate of biomass conversion but did not affect pellet size. The white fluorescent light contained several different wavelengths, and therefore, its effect on A. ficuum represents the cooperative effect of these wavelengths. Strong luminance of a white fluorescent light inhibited growth of A. ficuum mycelia on plates, whereas white LED light enhanced growth. The development of mycelia was also inhibited by blue LED light and enhanced by red LED light illumination. Investigating the effect of LED lights on the growth of A. ficuum could provide evidence on the luminous intensity that is sufficient for regulating fermentation by light.

Enhanced removal of dimethyl sulfide from a synthetic waste gas stream using a bioreactor inoculated with Microbacterium sp. NTUT26 and Pseudomonas putida.

The removal of dimethyl sulfide (DMS) from industrial gas streams has received a high priority due to its very low odorous threshold value and relatively low biodegradability compared to other reduced sulfur compounds. A variety of bacteria that utilize DMS as a carbon/energy source have been studied and the degradation pathway elucidated. However, to date, there have been few reports on the industrial application of such bacteria inoculated into a bioreactor for DMS treatment. An additional problem of such systems is the accumulation of intermediate metabolites that strongly impact on DMS removal by the microbe. The results reported here were obtained using a bioreactor inoculated with the H(2)S-degrader Pseudomonas putida and the DMS-degrader Microbacterium sp. NTUT26 to facilitate removal of metabolic intermediates and DMS. This bioreactor performed well (1.71 g-S/day/kg-dry packing material) in terms of DMS gas removal, based on an evaluation of the apparent kinetics and maximal removal capacity of the system. Under varying conditions (changes in start-up, inlet loading, shutdown, and re-start), the bioreactor inoculated with Microbacterium sp. NTUT26 and P. putida enhanced removal of high concentrations of DMS. Our results suggest that this type of bioreactor system has significant potential applications in treating (industrial) DMS gas streams.

Determination of Schistosoma japonicum circulating antigens in dilution serum by piezoelectric immunosensor and S/N enhancement.

A piezoelectric immunosensor assay was developed with immobilizing immunoglobulin G (IgG) as a probe to detect Schistosoma japonicum circulating antigens (SjCAg). Analytical strategy utilizes the polyclonal antibodies with broad-spectrum recognition to a complex target with high specificity. The immobilized antibodies were purified from immunized rabbit's sera (im-S) and infected rabbit's sera (inf-S) by S. japonicum. The detection capacities of antibodies were compared between the sera of different phenotypes and purified fractions. The sample dilution ratios were also evaluated and optimized. Additionally, the sera with a variety of infection degrees for validation could be discriminated quantitatively. The linear dose-response relationship indicates that the systematic sensitivity of this method is below 150 Hz and the lowest limit of detectable range is above 500 cercariae of S. japonicum infection for 2 weeks. The novel immunosensor technique is well potential to determine the SjCAg in serum samples for clinical diagnosis of parasitosis in early stage.