Functional enrichment analysis and disease enrichment analysis in three scRNA-seq datasets.

Functional enrichment analysis and disease enrichment analysis in three scRNA-seq datasets.</p

Isolation of the Novel Chiral Insecticide Paichongding (IPP) Degrading Strains and Biodegradation Pathways of <i>RR</i>/<i>SS</i>-IPP and <i>SR</i>/<i>RS</i>-IPP in an Aqueous System

Chiral insecticide paichongding (IPP) is a member of <i>cis</i>-nitromethylene neonicotinoid insecticides used in China. IPP was the promising replacement for imidacloprid as a result of its higher activity against imidacloprid-resistant insects. Two pairs of enantiomers, <i>RR</i>/<i>SS</i>-IPP and <i>SR</i>/<i>RS</i>-IPP, were separated by preparative high-performance liquid chromatography and employed in an aqueous system to investigate their biodegradation process. In this study, the strains G1-13/G1-14 and G2-19 with effective IPP degrading capability were isolated from agricultural soils. G1-14 was mutated from G1-13 by ultraviolet light exposure. Sequence alignment of 16S rRNA proved that these three strains belonged to the genus of Sphingobacterium. The degradation rate of <i>RR</i>/<i>SS</i>-IPP by Sphingobacterium sp. G1-13 and G1-14 reached 13 and 30% within 6 and 4 days, respectively. The degradation rate of <i>SR</i>/<i>RS</i>-IPP by Sphingobacterium sp. G2-19 could reach 35% within 5 days. Degradation intermediates (I1–I6) of enantiomers were detected, and two possible biodegradation pathways were proposed on the basis of the identification of metabolites

Expression of the <i>lux</i> genes in <i>Streptococcus pneumoniae</i> modulates pilus expression and virulence

<div><p>Bioluminescence has been harnessed for use in bacterial reporter systems and for <i>in vivo</i> imaging of infection in animal models. Strain Xen35, a bioluminescent derivative of <i>Streptococcus pneumoniae</i> serotype 4 strain TIGR4 was previously constructed for use for <i>in vivo</i> imaging of infections in animal models. We have shown that strain Xen35 is less virulent than its parent TIGR4 and that this is associated with the expression of the genes for bioluminescence. The expression of the <i>luxA-E</i> genes in the pneumococcus reduces virulence and down regulates the expression of the pneumococcal pilus.</p></div

Survival and weight loss of mice infected with T4P2.

<p>(A) Shows percentage survival of mice infected with TIGR4, Xen35 or T4P2 over time, statistical analysis was performed comparing T4P2 to TIGR4 or T4P2 to Xen35 using a logrank Test, * P<0.01. * above the strain indicated a statistical difference compared to T4P2. Only the survival group was used for analysis (n = 5 per bacterial strain) (B) Shows percentage weight loss of mice infected with Xen35, TIGR4 or T4P2. All groups were used for analysis; later groups have smaller numbers due to some mice being sacrificed. Statistical analysis was performed using a non-parametric Mann-Whitney two sample rank test, *P< 0.01 and **P<0.001.</p

Preliminary whole genome sequence changes in T4P2.

<p>Preliminary whole genome sequence changes in T4P2.</p

Survival and weight loss of mice infected with Xen35.

<p>(A) Percentage survival of mice intra-nasally inoculated with TIGR4 or Xen35 over time, statistical analysis was performed using a logrank Test, * P<0.01. * above the strain indicated a statistical difference compared to Xen35. Only the survival group was used for analysis (n = 5 per bacterial strain) (B) Percentage weight loss of mice infected with either Xen35 or TIGR4. All groups were used for analysis, later groups have smaller numbers due to some mice being sacrificed. Statistical analysis was performed using a non-parametric Mann-Whitney two sample rank test, *P<0.001.</p

Genome changes in Xen35 resulting in loss of function of the protein encoded or proteins containing large in frame deletions.

<p>Genome changes in Xen35 resulting in loss of function of the protein encoded or proteins containing large in frame deletions.</p

Bioluminescence of T4P strains.

<p>(A) Graph of bioluminescence of Xen35, TIGR4 and the T4P strains over time. Each point on the graph represents the average of a triplicate reading, Readings were taken every 20 minutes. (B) Western blotting analysis was performed on TIGR4, T4P1, P2, P3, P4 and T4Δ<i>rrgB</i> looking at RrgB protein expression in all strains (α-RrgB antibody). Equal protein loading was confirmed by equal expression of GroEL (α-GroEL antibody).</p

Energy production and consumption.

<p>(A) Reaction catalysed by luciferase enzyme (Flavin reductase) leading to light emission (bioluminescence). Reaction requires reduced riboflavin (FMNH₂), long chain aldehyde (RCHO) and oxygen (O₂) resulting in flavin mononucleotide (FMN), Water (H₂O), fatty acids (RCOOH) and light [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0189426#pone.0189426.ref003" target="_blank">3</a>] (B) Diagram showing adenosine triphosphate (ATP), reduced nicotinamide adenine dinucleotide (NADH), adenosine diphosphate (ADP) and oxidised nicotinamide adenine dinucleotide (NAD⁺) production and utilisation during glycolysis and pyruvate metabolism in <i>S</i>. <i>pneumoniae</i> [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0189426#pone.0189426.ref046" target="_blank">46</a>]. (C) Reaction catalysed by the fatty acid reductase complex required to produce the long chain aldehyde substrate for the luciferase enzyme. Reaction requires RCOOH, ATP, reduced nicotinamide adenine dinucleotide phosphate (NADPH) resulting in nicotinamide adenine dinucleotide phosphate (NADP), adenosine monophosphate (AMP), pyrophosphate (PPi) and RCHO [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0189426#pone.0189426.ref003" target="_blank">3</a>].</p

RT-PCR graph of <i>lux</i> and pilus genes expression in T4P2.

<p>Graph shows expression of <i>luxA-E</i> and the pilus islet genes (<i>rlrA</i>, <i>rrgA</i>, <i>rrgB</i>, <i>rrgC</i>, <i>srtB</i>, <i>srtC</i>, <i>srtD</i>) in T4P2. Fold change represents that of T4P2 compared to Xen35. Each bar represents the average of at least two replicas and error bars the standard deviation.</p