Dendrogram combining PFGE patterns of XbaI-digested DNA from the 76 isolated <i>Cronobacter</i> spp.

<p>Isolate information including GenBank accession No., biogroups, isolation years, and manufacturer. * Three manufacturers shared the genetic subtypes MJ1150, WJ1151, and WJ1163; & Species isolates from the same batch showed different genetic subtypes (WJ9197 and WJ 9199); # A single batch of rice powder from manufacturer A contained two species (WJ0857, WJ0858, and WJ 8060—<i>C</i>. <i>sakazakii</i>; WJ8059—<i>C</i>. <i>dublinensis</i>); $ the same PFGE-pattern in batches produced a year apart, implying ongoing contamination of the rice product (WJ9197 and WJ1166).</p

<i>Cronobacter</i> spp. isolated from dehydrated rice powder (DRP) detected by culture and 16S rRNA gene PCR amplification and sequencing.

<p><i>Cronobacter</i> spp. isolated from dehydrated rice powder (DRP) detected by culture and 16S rRNA gene PCR amplification and sequencing.</p

Maximum likelihood tree of the 76 isolated <i>Cronobacter</i> and outgroup species <i>Citrobacter koseri</i> based on the fusA alleles (438 bp) of the <i>Cronobacter</i> multilocus sequence typing dataset.

<p>This tree was generated using the MEGA (v. 6.06) with 1000 bootstrap replicates.</p

Additional file 2: Figure S2. of In vitro activity of ten essential oils against Sarcoptes scabiei

The mites in fumigation bioassay observed under a stereomicroscope. The mites stayed firmly attached to the bottom of the Petri dish which has been turned over. (JPG 400 kb

Additional file 1: Figure S1. of In vitro activity of ten essential oils against Sarcoptes scabiei

Thousands of mites on the crusts collected from the ear canal of the pig model. (JPG 639 kb

Additional file 1: Figure S1. of Population structure of Haemonchus contortus from seven geographical regions in China, determined on the basis of microsatellite markers

Locations of seven populations of Haemonchus contortus in China. (TIF 2529 kb

Bone Marrow-Derived Mesenchymal Stem Cells Maintain the Resting Phenotype of Microglia and Inhibit Microglial Activation

<div><p>Many studies have shown that microglia in the activated state may be neurotoxic. It has been proven that uncontrolled or over-activated microglia play an important role in many neurodegenerative disorders. Bone marrow-derived mesenchymal stem cells (BMSCs) have been shown in many animal models to have a therapeutic effect on neural damage. Such a therapeutic effect is attributed to the fact that BMSCs have the ability to differentiate into neurons and to produce trophic factors, but there is little information available in the literature concerning whether BMSCs play a therapeutic role by affecting microglial activity. In this study, we triggered an inflammatory response situation <i>in vitro</i> by stimulating microglia with the bacterial endotoxin lipopolysaccharide (LPS), and then culturing these microglia with BMSC-conditioned medium (BMSC-CM). We found that BMSC-CM significantly inhibited proliferation and secretion of pro-inflammatory factors by activated microglia. Furthermore, we found that the phagocytic capacity of microglia was also inhibited by BMSC-CM. Finally, we investigated whether the induction of apoptosis and the production of nitric oxide (NO) were involved in the inhibition of microglial activation. We found that BMSC-CM significantly induced apoptosis of microglia, while no apoptosis was apparent in the LPS-stimulated microglia. Our study also provides evidence that NO participates in the inhibitory effect of BMSCs. Our experimental results provide evidence that BMSCs have the ability to maintain the resting phenotype of microglia or to control microglial activation through their production of several factors, indicating that BMSCs could be a promising therapeutic tool for treatment of diseases associated with microglial activation.</p></div

Apoptosis assessed by annexin V/propidium iodide (PI) assay.

<p>Annexin V/PI staining was performed to assess the apoptosis rate in microglia. After incubating in DMEM/F-12 or BMSC-CM with or without LPS (1 µg/mL) for 48 hours, microglia were harvested. Cells were then stained with annexin V-FITC and propidium iodide and analyzed by flow cytometry. The results are presented as density plots of propidium iodide vs. annexin V-FITC. Apoptotic cells have high annexin V-FITC and low propidium iodide staining (lower-right quadrant). Numbers on the graph represent the percentages of apoptotic cells in the different groups.</p

BMSCs inhibit phagocytosis by microglial cells.

<p>We tested the capacity of microglia to phagocytose fluorescent-labeled latex beads by observing them under a fluorescence microscope (magnification 400×). The microglial cells were incubated in DMEM/F-12 or conditioned medium with or without LPS (1 µg/mL) at 37°C in 5% CO2 in air and 95% humidity. The nuclei were counterstained with DAPI. Following incubation with LPS (1 µg/mL) for 4 hours, microglia clearly ingested more latex beads than untreated control groups, while the phagocytic activity of microglia in the conditioned medium (CM) treated-groups was significantly inhibited.</p

Role of NO in the BMSC-mediated inhibition of microglial proliferation.

<p>We cultured microglia in conditioned medium derived from BMSCs with or without inhibition of NO production. In the CM+SMT groups, microglia were incubated in conditioned medium derived from BMSCs in which NO production was inhibited by the NO inhibitor S-methylisothiourea sulfate (SMT, 1 mM). We found that cells cultured in CM after inhibition of NO production (CM+SMT groups) showed increased proliferation compared to cells cultured in CM without inhibition of NO production (CM groups). Bars represent means plus or minus SD obtained from six independent experiments. A P value of less than 0.05 (*), or less than 0.01 (**) was considered statistically significant.</p