Mycoplasma genitalium Lipoproteins Induce Human Monocytic Cell Expression of Proinflammatory Cytokines and Apoptosis by Activating Nuclear Factor κB

This study was designed to investigate the molecular mechanisms responsible for the induction of proinflammatory cytokines gene expression and apoptosis in human monocytic cell line THP-1 stimulated by lipoproteins (LPs) prepared from Mycoplasma genitalium. Cultured cells were stimulated with M. genitalium LP to analyze the production of proinflammatory cytokines and expression of their mRNA by ELISA and RT-PCR, respectively. Cell apoptosis was also detected by Annexin V-FITC-propidium iodide (PI) staining and acridine orange (AO)-ethidium bromide (EB) staining. The DNA-binding activity of nuclear factor-κB (NF-κB) was assessed by electrophoretic mobility shift assay (EMSA). Results showed that LP stimulated THP-1 cells to produce tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 in a dose-dependent manner. The mRNA levels were also upregulated in response to LP stimulation. LPs were also found to increase the DNA-binding activity of NF-κB, a possible mechanism for the induction of cytokine mRNA expression and the cell apoptosis. These effects were abrogated by PDTC, an inhibitor of NF-κB. Our results indicate that M. genitalium-derived LP may be an important etiological factor of certain diseases due to the ability of LP to produce proinflammatory cytokines and induction of apoptosis, which is probably mediated through the activation of NF-κB

Draft genome sequence of the mulberry tree Morus notabilis

Human utilization of the mulberry–silkworm interaction started at least 5,000 years ago and greatly influenced world history through the Silk Road. Complementing the silkworm genome sequence, here we describe the genome of a mulberry species Morus notabilis. In the 330-Mb genome assembly, we identify 128 Mb of repetitive sequences and 29,338 genes, 60.8% of which are supported by transcriptome sequencing. Mulberry gene sequences appear to evolve ~3 times faster than other Rosales, perhaps facilitating the species’ spread worldwide. The mulberry tree is among a few eudicots but several Rosales that have not preserved genome duplications in more than 100 million years; however, a neopolyploid series found in the mulberry tree and several others suggest that new duplications may confer benefits. Five predicted mulberry miRNAs are found in the haemolymph and silk glands of the silkworm, suggesting interactions at molecular levels in the plant–herbivore relationship. The identification and analyses of mulberry genes involved in diversifying selection, resistance and protease inhibitor expressed in the laticifers will accelerate the improvement of mulberry plants

Evaluating the gross pathology in the genital tracts of CBA/j mice infected with <i>C</i>. <i>muridarum</i>.

<p>The three parallel groups of CBA/J mice infected with luciferase-expressing <i>C</i>. <i>muridarum</i> intragastrically (i.g., panel a, n = 5), intrarectally (i.r., b, n = 5) or intravaginally (i.vag., panels c, n = 5) as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155880#pone.0155880.g003" target="_blank">Fig 3</a> legend were sacrificed on day 70 after infection for visually identifying (arrow) and scoring (white numbers) hydrosalpinx. Mice with hydrosalpinx on either side of the genital tract were defined positive for hydrosalpinx. The severity of hydrosalpinx was scored from each oviduct independently and the scores from both sides of the same mouse were added together as the score for that mouse. Both incidence rates and severity scores of hydrosalpinx from each group were listed beside the corresponding images. Note that significant hydrosalpinx was detected only in mice infected intravaginally but intragastrically or intrarectally.</p

Molecular Analysis of <i>Staphylococcus epidermidis</i> Strains Isolated from Community and Hospital Environments in China

<div><p>Background</p><p><i>Staphylococcus epidermidis</i> is a common cause of nosocomial infections worldwide. This study analyzed the differences in genetic endowment and clonal lineages with pathogenesis and resistance traits of <i>S. epidermidis</i> isolates collected from community and hospital environments (patients and healthcare staff) of the same ecological niche, time period, and geographical location in China.</p><p>Methodology/Principal Findings</p><p>Molecular epidemiology and population analysis showed that nasal colonization rates of <i>S. epidermidis</i> in the community of Shanghai area of China and in healthcare personnel were 44.8% (methicillin-resistant <i>S. epidermidis</i>, MRSE: 17.2%) and 61.3% (MRSE: 30.0%), respectively. 86.7% of clinical isolates were MRSE. Among the strains studied, 44 sequence types (STs) were identified with 91.7% belonging to clonal complex 2 (CC2). Only 40.8% isolates from patients were also found in healthy individuals. MRSE-ST2-SCC<i>mec</i>III was the predominant clone in clinical isolates, almost resistant to all antibiotics tested. Biofilm-related genes <i>IS256</i> and <i>icaA</i> were detected in majority of the predominant clinical MRSE-ST2 clone with a 40.5% biofilm-positive rate. No ST2 isolate was found in community setting. We found a high prevalence of arginine catabolic mobile element (ACME) (74.1%). The prevalence of ACME-<i>arc</i> and ACME-<i>opp3</i> clusters was 71.6% and 32.4%, respectively. Methicillin-sensitive <i>S. epidermidis</i> (MSSE) isolates harbored more ACME (83.3%) than MRSE isolates (67.7%), and there was no association between ACME and SCC<i>mec</i> types. An association was found between low-level ACME presence and invasive infections.</p><p>Conclusions/Significance</p><p>We observed a high level of diversity within <i>S. epidermidis</i> in this study, with CC2 as the dominant clonal complex in both community and hospital settings. Only 40.8% of the isolates from patients were also found in healthy individuals. Contrary to that biofilm formation and multiple antibiotic resistance were associated closely with pathogenicity of <i>S. epidermidis</i>, ACME was more likely to be an indicator for colonization rather than a virulence factor.</p></div

Monitoring <i>C</i>. <i>muridarum</i> infection in the GI and genital tracts of C57BL/6J and C3H/HeJ mice following an intrarectal inoculation.

<p>C57BL/6J (panels a1-a3, n = 4) and C3H/HeJ (b1-3, n = 3) mice were intrarectally infected with <i>C</i>. <i>muridarum</i> and both anorectal and cervico-vaginal swabs were taken on different days after infection as listed along the X-axis in the bottom of the figure. The number of live organisms recovered from the swabs was expressed as Log10 IFUs. "ND” denotes not detected. On day 70 after infection, mice were sacrificed for harvesting the genital tracts for visually identifying and scoring hydrosalpinx (a3 & b3) as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155880#pone.0155880.g003" target="_blank">Fig 3</a> legend. Note that although significant levels of live organisms were continuously detectable in the mouse GI tract throughout the experiment, no significant level of live organisms was detected in the mouse vaginal swabs and there was no significant hydrosalpinx in any of the mice.</p

Evaluating the microscopic pathology in the mouse genital tracts.

<p>The same mouse genital tissues harvested on day 70 after infection as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155880#pone.0155880.g005" target="_blank">Fig 5</a> legend were examined for inflammatory infiltration in the genital tract tissues. (A) Representative images of H&E-stained sections of vagina (panels a, d & g under 10x objective lens while a1, d1 & g1 under 100x), uterine/uterine horn (b, e & h, 10x; b1, e1 & h1, 100x) and oviduct (c, f & i, 10x; c1, f1 & i1, 100x) from mice infected intragastrically (i.g., a-c), intrarectally (i.r., d-f) and intravaginally (i.vag., g-i) were shown. (B) The Inflammatory infiltration from the vagina (a), uterine/uterine horn (b) and oviduct (c) tissues were semi-quantitatively scored (shown along the Y-axis) as described in the Materials and Method section. Kruskal-Wallis Test was used to calculate the p values listed in the figure. Note that only the tissues from the intravaginally infected mice displayed significant levels of inflammatory infiltration.</p

Recovering live <i>C</i>. <i>muridarum</i> organisms from GI versus genital tracts of CBA/J mice.

<p>The same two groups of mice as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0155880#pone.0155880.g001" target="_blank">Fig 1</a> legend and an additional group of CBA/J (n = 5) infected intravaginally (i.vag.) with the same amount of luciferase-expressing <i>C</i>. <i>muridarum</i> were monitored for live chlamydial organism shedding in their rectal (panels a, c & e) and vaginal (b, d & f) swabs on different days after infection as indicated along the X-axis at the bottom of the figure. The results were expressed as Log10 IFUs displayed along the Y-axis. Note that no significant live organisms were detected in the vaginal swabs of mice infected in the GI tract throughout the experiments.</p

<i>In vivo</i> imaging of mice infected intragastrically or intrarectally with luciferase-expressing <i>C</i>. <i>muridarum</i>.

<p>CBA/J mice were intragastrically (i.g., n = 5, panels a-i) or intrarectally (i.r., n = 5, j-r) inoculated with 5 x 10⁴ IFUs of luciferase-expressing <i>C</i>. <i>muridarum</i>. On different days after the inoculations as indicated on top of the figure, a whole body <i>in vivo</i> imaging was used to detect the luciferase-generated bioluminescence signals as displayed in red/green/blue colors (in the order of decreasing intensity). Images taken from one of the mice in each group were shown. It is worth noting that the bioluminescence signals were detectable as early as day 3 after inoculation at the initial inoculation sites either stomach (panel a) or anorectum (j). Starting on day 7, the signals persisted in the mouse abdominal area throughout the experiments with similar distribution patterns regardless of the initial inoculation routes.</p

Monitoring <i>C</i>. <i>muridarum</i> infection in both the GI and genital tracts following an intrarectal inoculation with 1 x 10⁷ IFUs.

<p>CBA/J mice were intrarectally (n = 4) inoculated with 1 x 10⁷ IFUs of luciferase-expressing <i>C</i>. <i>muridarum</i>. (A) On different days after the inoculations as indicated on top of the figure, a whole body <i>in vivo</i> imaging was used to detect the luciferase-generated bioluminescence signals as displayed in red/green/blue colors (in the order of decreasing intensity). Images taken from one of the mice in each group were shown. (B) At the same time, these mice were monitored for live chlamydial organism shedding in their rectal (panel a) and vaginal (b) swabs on different days after infection as indicated along the X-axis at the bottom of the figure. The results were expressed as Log10 IFUs displayed along the Y-axis. Note that no significant live organisms were detected in the vaginal swabs throughout the experiment.</p