Because of their small genomes, mycoplasmas are commonly chosen as model organisms for the study of the minimal gene set needed to support the growth of free-living bacteria and are ideal for dissecting the role of individual genes in pathogenesis. The most comprehensive work has been performed with Mycoplasma genitalium. Using transposon mutagenesis, 387 of the 480 protein-coding regions and all of the 37 genes coding for RNA species were identified Selleckchem BIBW2992 as essential for the growth of M. genitalium (Glass et al., 2006). Because of the lack of gene duplication and the absence of redundant pathways,

it should be expected that the 580-kb genome of M. genitalium would have more essential genes than a bacterium with a large genome such as Bacillus subtilis, a Gram-positive bacterium to which the mycoplasmas are phylogenetically related. Indeed, only 271 of the 4100 genes of B. subtilis are thought to be essential (Kobayashi et al., 2003). Mycoplasma pulmonis is a pathogen of rats and mice and the etiological agent of murine respiratory mycoplasmosis (Lindsey & Cassell, 1973). In an earlier study of a transposon library of M. pulmonis, 321 of the 782 protein-coding regions were identified as dispensable for growth (French et al., 2008). The criteria used to consider a gene to be inactivated were that at least 10% of the coding region from the annotated 5′ start site or at least 15% of the coding region from the 3′ end would be

truncated. The size of the library was large enough to conclude that nonessential genes >1 kb were inactivated. The previous studies relied on Tn4001T, which transposes actively in the mycoplasmal Crizotinib genome. A more recent study in Mycoplasma arthritidis made use of the Tn4001TF1 minitransposon, derived from Tn4001 (Dybvig et al., 2008). The minitransposon readily transposed into the genome, but

once inserted, was unable to undergo subsequent Tryptophan synthase transposition events. This minitransposon was applied to M. pulmonis in the current study. The use of the minitransposon generated a superior library with inactivation of 39 genes that were previously thought to be essential. Mycoplasma pulmonis strain CT (Davis et al., 1986) was propagated in mycoplasma broth (MB) and mycoplasma agar (MA) as described (Dybvig et al., 2000; Simmons & Dybvig, 2003). CT was transformed with plasmid pTF85, carrying minitransposon Tn4001TF1 (Dybvig et al., 2008), using 36% polyethylene glycol and selecting for resistance to tetracycline as described (Dybvig et al., 2000). Individual colonies were picked, grown in 1 mL MB and stored at −80 °C as described (French et al., 2008). The genomic location of the transposon was determined for each library member by DNA sequence analysis of an inverse PCR product containing the junction between the transposon and the adjacent mycoplasmal DNA using primers and reaction conditions as described (Teachman et al., 2002; French et al., 2008).

Biofilm EPS can contain polysaccharides, proteins, nucleic acids (Flemming & Wingender, 2002), but few specific reports exist on the EPS matrix of P. putida biofilms. As a first approach to address the increased biofilm formation by the TOL-carrying strain, we attempted to extract EPS from biofilm-containing stagnant liquid cultures by a standard protocol using a cation-exchange resin (Frølund et al., 1996). However, it was not possible to extract the EPS component that caused the higher viscosity: both extracts had similar low viscosities [KT2440: 1.3 cSt Pexidartinib in vivo vs. KT2440 (TOL) 1.8 cSt], and extractable carbohydrate and nucleic acid contents were similar for both strains (Table 3).

This suggested that a major part of the EPS was not extractable and remained cell associated. Further attempts to increase Forskolin extraction intensities rapidly caused cell lysis, as observed by rRNA release in extracts (results not shown). Consequently, we attempted to analyze EPS components directly in the biofilm-containing liquid cultures. Total carbohydrate contents were about double as high as in the extracts, but no striking differences between the two strains were found. Total DNA contents of the two cultures were, however, clearly different: The TOL-carrying strain contained twice the amount of total DNA, as compared with the respective plasmid-free cultures, as similar cellular biomass contents measured as particulate protein. The next approach

involved direct visualization by microscopy. To check for the differential presence of eDNA in liquid–solid interface biofilms, 7-day-old flow cell biofilms of the TOL-free or TOL-carrying KT2440 were stained with propidium iodide (PI). Diffuse PI fluorescence

was colocalized with the larger (presumably older) microcolonies formed by the TOL-carrying strain, indicating that eDNA was a major Cobimetinib price constituent (Fig. 1). The plasmid-free strains, again, did not form thick biofilms, and eDNA was not observed. Using similar approaches, eDNA has been observed in various pure-culture biofilms (e.g. Pseudomonas aeruginosa, Bacillus subtilis, Enterococcus faecalis, environmental isolate) (Whitchurch et al., 2002; Bockelmann et al., 2006; Thomas et al., 2009; Vilain et al., 2009), and eDNA is, therefore, increasingly being considered a potential core element of the biofilm matrix. eDNA has similarly been observed in flocs and unsaturated biofilms of environmental Pseudomonas (Palmgren & Nielsen, 1996; Steinberger & Holden, 2005). In contrast to our observations, this eDNA remained easily extractable by chemical or thermal treatment methods isolates (Palmgren & Nielsen, 1996; Steinberger & Holden, 2005). Ultimately, eDNA was successfully extracted from TOL-free and TOL-carrying KT2440 cultures by enzymatic treatment using cellulase and proteinase K followed by centrifugation (Wu & Xi, 2009), without concurrent increase in cell lysis as ascertained using live/dead cell staining.

Based on observational studies [6,15–19] and expert opinion, current US guidelines recommend immunization of patients with PPV-23 when CD4 counts are above 200 cells/μL [20], whereas the World Health Organization (WHO) states that the pneumococcal polysaccharide vaccine may be considered for people with HIV infection in WHO clinical stage 1 or, if CD4 testing is available, with a CD4 count above 500 cells/μL [21]. However, study quality and the risk of bias in these studies have not been assessed. Following the recent success of 7-valent pneumococcal conjugate vaccine in preventing vaccine serotype-specific IPD in a cohort consisting primarily of HIV-infected Malawian adolescents [22],

a critical evaluation of PPV-23 effectiveness is needed. Immunity against capsulated bacteria such as pneumococci AZD2281 manufacturer depends on the formation of opsonic antibodies, which can be produced by B cells in response to polysaccharide stimulation. These antigens are classified as T-cell independent type 2 (TI-2) antigens, as they active B cells directly without assistance from T

cells [23]. Untreated HIV-infected subjects buy Nintedanib have reduced antibody responses to PPV-23 [8], which correlate with falling CD4 cell counts [24,25]. HAART partially restores the immune system by reducing HIV replication and immune activation, and improving CD4 cell counts. However, certain abnormalities of the immune system persist even years after HAART initiation, including a low CD4:CD8 ratio, a low naïve:memory cell ratio, expansion of CD28 effector T cells and a reduced T cell repertoire [26]. HAART may affect qualitative aspects of the PPV-23 response by restoring the expression of certain genes used in the PPV-23 response to normal levels and by improving the specific immunoglobulin G response to vaccines, including PPV-23 [11–13,27,28]. Thus, when assessing PPV-23 effectiveness in persons with HIV infection, it

selleckchem should be borne in mind that the effects of CD4 cell count and HAART treatment may be important. A number of risk factors for pneumococcal disease have been identified over the past 20 years (Table 1). Awareness of these risk factors is critical in the assessment of PPV-23 studies because, unless adjusted for in the statistical analysis, any risk factor for pneumococcal disease may potentially confound the measurement of vaccine effectiveness. CD4 cell count is an example of a well-known risk factor for pneumonia and pneumococcal disease [4,6,16,17,29–31]. Other risk factors related to progression of HIV infection are HIV RNA [4,30,32,33] and clinical disease stage [16,17,29]. The aim of this review was to systematically identify and critically assess all peer-reviewed and non-peer-reviewed literature on observational studies and clinical trials of the effectiveness of PPV-23 in terms of clinical endpoints in HIV-infected adults.

In the case of the latter drug, it may be particularly appropriate for use in the obese Omipalisib solubility dmso subject with GDM. “
“Structured education programmes support and enable people with diabetes to develop self-management skills. Insulin management central to DAFNE is restricted to those with type 1 diabetes of at least six months’ duration and on

multiple dose regimens. DESMOND is available for all with type 2 diabetes but does not include guidance on how to self-manage diabetes with insulin. Our aims were to develop an education programme for people with type 1 or 2 diabetes already on insulin and who may be using a variety of insulin regimens, to enable effective self-management, improve confidence, reduce hypoglycaemia and enable peer group support. An evidence-based curriculum, developed in line with NICE principles, was piloted. This consisted of three half-day Sirolimus ic50 sessions held during a one-month period with up to 10 participants and supporters invited to attend. Four further programmes were held; education was tailored to the individual needs of groups and verbal evaluation was undertaken. Anonymised patient satisfaction questionnaires

were posted at programme completion. Audit included clinical data, demographics, patient satisfaction and health care professional assessment of content. There were 40 participants Etoposide datasheet over five courses; 20% (n=8) were type 1, 68% (n=27) were male, average age was 58 years (range 35–82 years), and 55% were South Asian (n=22). In 38 of 40 participants where a recorded pre- and three months post-intervention was available, an average HbA1c reduction of 1.18% was achieved – i.e. 9.02% reduced to 7.84% (75.1mmol/mol

reduced to 62.2mmol/mol). Twenty-five participants (62.5%) returned the survey form: 96% (24/25) said diabetes control improved, and all felt more confident to adjust insulin; 96% (23/24) felt more confident to treat ‘hypos’ (one stated ‘hypos’ had not reduced) and 96% (24/25) felt they learned more by attending the programme. This programme met participants’ individual needs, increased confidence in insulin management and improved glycaemic control in a high ethnic mix poulation. Copyright © 2014 John Wiley & Sons. Practical Diabetes 2014; 31(2): 54–57 “
“The central theme of this article is that a person with diabetes who thinks they are ‘not good enough’ at diabetes self-management is manifesting a sense of shame. This fundamental human attribute is often the most significant, underlying issue that people face in psychotherapy and yet neither the ICD-10 nor the DSM-V recognises shame as a discrete diagnosis.

This study

investigated the effect of trace iron conditions on the growth of these two species, as well as their response to iron sequestration by chelators. Metal analysis by ICP-MS revealed high residual concentrations (0.12 μM) of iron in the chemically defined medium used in spite of the absence of added iron and demonstrated the requirement for deferration and confirmatory trace Fe analysis. The iron contamination from other medium constituents could be successfully reduced to <0.02 μM in batch processes using an insoluble chelating resin. Cu concentrations were also significantly reduced in the extracted chemically defined medium, but significant recovery of growth could be see more achieved by supplementation of the extracted medium with Fe only. Both C. albicans and C. vini were found to require Cytoskeletal Signaling inhibitor approximately 0.5 μM added iron for complete unrestricted growth in the extracted chemically defined medium, but differed in their abilities to grow at reduced iron concentrations. The observed differences between C. albicans and C. vini were consistent with the different environments these

respective yeast species typically colonize or invade. Grape musts and wines, in which C. vini typically appears as a spoilage yeast, generally have high Fe concentrations of between 30 and 200 μM (Ough et al., 1982). The predominantly reducing environment and the low pH of grape musts and wines also favour the formation of the more soluble free ferrous species, and this Fe would be expected to have a higher bioavailability (Howard,

1999). In sharp contrast, the ecological niches that C. albicans can colonize or invade in relation to human pathogenesis are highly limiting for Fe (Weinberg, 1999). Desferrioxamine and deferiprone are two chelators used clinically very to relieve the Fe overload associated with certain human haematological disorders such as thalassaemia (Chaston & Richardson, 2003; Franchini, 2006). Desferrioxamine failed to inhibit both C. albicans and C. vini. Deferiprone did not inhibit C. vini while leading to a slightly increased lag phase in C. albicans. However, the observed differences between C. albicans and C. vini persisted in their growth response in the presence of lactoferrin. Lactoferrin is a major component of the mammalian innate immune system (Actor et al., 2009) and one of the vertebrate host defence Fe chelators, which is present in mucosal secretions (Gonzalez-Chavez et al., 2009). Lactoferrin, at the physiologically relevant concentration of 0.25 mg mL−1 and at pH 4.5, and thus, representative of the vaginal environment (Novak et al., 2007), only led to a transient inhibition of C. albicans, but inhibited the growth of C. vini over the incubation period. The results are in agreement with the lack of observed pathogenicity of C. vini and its greater susceptibility to iron restriction, while the pathogenicity of C.

This study

investigated the effect of trace iron conditions on the growth of these two species, as well as their response to iron sequestration by chelators. Metal analysis by ICP-MS revealed high residual concentrations (0.12 μM) of iron in the chemically defined medium used in spite of the absence of added iron and demonstrated the requirement for deferration and confirmatory trace Fe analysis. The iron contamination from other medium constituents could be successfully reduced to <0.02 μM in batch processes using an insoluble chelating resin. Cu concentrations were also significantly reduced in the extracted chemically defined medium, but significant recovery of growth could be selleck chemical achieved by supplementation of the extracted medium with Fe only. Both C. albicans and C. vini were found to require DZNeP ic50 approximately 0.5 μM added iron for complete unrestricted growth in the extracted chemically defined medium, but differed in their abilities to grow at reduced iron concentrations. The observed differences between C. albicans and C. vini were consistent with the different environments these

respective yeast species typically colonize or invade. Grape musts and wines, in which C. vini typically appears as a spoilage yeast, generally have high Fe concentrations of between 30 and 200 μM (Ough et al., 1982). The predominantly reducing environment and the low pH of grape musts and wines also favour the formation of the more soluble free ferrous species, and this Fe would be expected to have a higher bioavailability (Howard,

1999). In sharp contrast, the ecological niches that C. albicans can colonize or invade in relation to human pathogenesis are highly limiting for Fe (Weinberg, 1999). Desferrioxamine and deferiprone are two chelators used clinically Urease to relieve the Fe overload associated with certain human haematological disorders such as thalassaemia (Chaston & Richardson, 2003; Franchini, 2006). Desferrioxamine failed to inhibit both C. albicans and C. vini. Deferiprone did not inhibit C. vini while leading to a slightly increased lag phase in C. albicans. However, the observed differences between C. albicans and C. vini persisted in their growth response in the presence of lactoferrin. Lactoferrin is a major component of the mammalian innate immune system (Actor et al., 2009) and one of the vertebrate host defence Fe chelators, which is present in mucosal secretions (Gonzalez-Chavez et al., 2009). Lactoferrin, at the physiologically relevant concentration of 0.25 mg mL−1 and at pH 4.5, and thus, representative of the vaginal environment (Novak et al., 2007), only led to a transient inhibition of C. albicans, but inhibited the growth of C. vini over the incubation period. The results are in agreement with the lack of observed pathogenicity of C. vini and its greater susceptibility to iron restriction, while the pathogenicity of C.

The regulatory mechanism for PHB biosynthesis in B. thuringiensis and diverse Bacillus species is still poorly understood. We now report that disruption of the sigH gene or the gene encoding the master sporulation transcription factor Spo0A severely impaired PHB accumulation in B. thuringiensis. Complementation

of the spo0A mutation with the spo0A gene restored PHB accumulation. We have found that the requirement of Spo0A for PHB accumulation is independent of the transition state regulator AbrB and of loss of sporulation ability. We also show that Spo0A is required for the expression of three genes involved in PHB biosynthesis. These findings have uncovered a new role of Spo0A in the regulation of stationary-phase-associated cellular events. Poly(3-hydroxybutyrate) (PHB) is a polyester accumulated by numerous bacteria as an intracellular carbon and energy storage Selleckchem GDC-0068 material in response to nutritional stress (for recent reviews, see Waltermann & Steinbuchel, 2005; Jendrossek, 2009). A cluster of five PHB-related genes, that is phaP (encoding a phasin protein),

phaQ (a repressor of phaP expression), phaR (a subunit of PHB synthase), phaB (acetoacetyl-CoA reductase), and phaC (a subunit of PHB synthase), was previously identified and characterized from the PHB-producing Bacillus megaterium (McCool & Cannon, 1999; Lee et al., 2004). The PHB synthase of B. megaterium requires both the AP24534 clinical trial PhaR subunit and the PhaC subunit for activity (McCool & Cannon, 2001). The genetic organization of Bacillus thuringiensis counterparts of these five PHB-related genes is the same as that of B. megaterium. However, the regulatory mechanism for PHB biosynthesis

in diverse Bacillus species is poorly understood. The master transcription factor Spo0A is a response regulator that plays a central role in the initiation of sporulation of Bacillus subtilis and other Bacillus species (for recent reviews, see Piggot Adenosine & Hilbert, 2004; Stephenson & Lewis, 2005). Bacillus subtilis Spo0A is also known to be involved in controlling other cellular events, such as biofilm formation (Hamon & Lazazzera, 2001), development of competence for DNA uptake (Hahn et al., 1995), cannibalism (Gonzalez-Pastor et al., 2003), cold and heat resistance (Mendez et al., 2004), bacilysin biosynthesis (Karatas et al., 2003), and extracellular-degradative enzyme production (Kodama et al., 2007). Phosphorylated Spo0A is capable of binding to a specific DNA sequence, called the 0A box (5′-TGNCGAA-3′), found in promoter regions of its target genes to repress or stimulate transcription (Molle et al., 2003). The pathway to Spo0A phosphorylation involves a multicomponent phosphorelay, in which the phosphate group is initially transferred from multiple sensor histidine kinases to Spo0F, then to Spo0B, and eventually to Spo0A.

While overall tone-evoked response magnitudes were comparable between the two structures, tone signal : noise was significantly greater within the OT than in the PCX. Panobinostat research buy No effect of tone frequency (1–55 kHz) was found within either structure, with most units being narrowly tuned to a single frequency. These results suggest that a major portion of odor-evoked output from the olfactory bulb (i.e. that entering the OT and PCX) is subject to auditory sensory input in a manner that may modulate odor information processing,

odor-guided behaviors and perception. “
“Behavioral rhythms induced by methamphetamine (MAP) and daily restricted feeding (RF) in rats are independent of the circadian pacemaker in the suprachiasmatic nucleus (SCN), and have been regarded to share a common oscillatory mechanism. In the present study, in order to examine the responses of brain oscillatory systems to MAP and RF, circadian rhythms in clock gene, Period2, expression were measured in several brain ALK inhibitor areas in rats. Transgenic rats carrying a bioluminescence reporter of Period2-dLuciferase were subjected to either daily injection

of MAP or RF of 2 h at a fixed time of day for 14 days. As a result, spontaneous movement and wheel-running activity were greatly enhanced following MAP injection and prior to daily meal under RF. Circadian Per2 rhythms were measured in the cultured brain tissues containing one of the following structures: the olfactory bulb; caudate-putamen; parietal cortex; substantia nigra; and SCN. Except for the SCN, the circadian Per2 rhythms in the brain tissues were significantly phase-delayed by 1.9 h on average in MAP-injected rats as compared with the saline-controls. On the other hand, the circadian rhythms outside the SCN were significantly phase-advanced by 6.3 h on average in rats under RF as compared with those under ad libitum feeding. These findings indicate that the circadian rhythms in specific brain areas of the central dopaminergic system respond differentially to MAP

injection and RF, suggesting that different oscillatory mechanisms in the brain underlie the MAP-induced behavior and pre-feeding activity under RF. “
“Glutamate is the major excitatory neurotransmitter of the central nervous system in vertebrates. Excitotoxicity, caused by over-stimulation why of the glutamate receptors, is a major cause of neuron death in several brain diseases, including epilepsy. We describe here how behavioural seizures can be triggered in adult zebrafish by the administration of kainate and are very similar to those observed in rodent models. Kainate induced a dose-dependent sequence of behavioural changes culminating in clonus-like convulsions. Behavioural seizures were suppressed by DNQX (6,7-dinitroquinoxaline-2,3-dione) dose-dependently, whilst MK-801 (a non-competitive NMDA receptor antagonist) had a lesser effect.

In addition, we performed receiver operating characteristic (ROC) analysis to assess the accuracy of EBV DNA load as a predictive marker of lymphoma [as estimated by the area under the curve (AUC)]. The optimal cut-off value of EBV DNA load for differentiating patients at risk of lymphoma from other patients was determined as the point of the ROC curve with the shortest distance to the 100/100% sensitivity/specificity angle (upper left corner) [i.e. lowest value for the term (1 – sensitivity)2 + (1 – specificity)2, assuming equal costs of false positive and false negative results]. The sensitivity, specificity and OR for developing lymphoma were then provided for the identified

cut-off point. All statistical analyses were performed using sas 9.2 (SAS Institute Inc., VEGFR inhibitor Cary, NC). EBV DNA was positive in PBMC samples from all lymphoma cases collected over the 3 years preceding the Obeticholic Acid diagnosis, while it was positive in 78 to 81% of samples from controls collected during the same period of time (Fig. 1a). Interestingly, eight of the 37 controls had undetectable EBV loads in PBMC1 while none of the 20 cases had undetectable EBV loads in PBMC1 (P = 0.04) (Fig. 1a). EBV load in PBMCs measured a median of 10 months before diagnosis was associated with an increased risk of B lymphoma [OR 2.48 (95% CI 1.16; 5.32) per increase in EBV

load of 1 log copies/106 PBMCs] (Table 2). Similar results were obtained when the OR was adjusted for CD4 cell count nadir instead of CD4 cell count at sample date (OR 2.33; 95% CI 1.12; 4.81). The OR associated with EBV load quantified in a sample collected earlier (median of 24 months before diagnosis) was of borderline significance, probably because of a smaller number of PBMC samples available for that period. When we restricted the analysis to the patients with a CD4 cell count > 300 cells/μL, the median EBV load was still lower in controls (median 2.69) than in cases (median 3.63), mainly because four out of 14 controls had undetectable EBV load vs. none of

seven cases. EBV DNA was more often detectable (> to EBV PCR threshold value or detectable but < to EBV PCR threshold value) in sera from cases than in sera from controls (with 24 to 25% positive detection in the last 3 years for cases vs. 8 to 10.5% for Unoprostone controls) (Fig. 1b); however, this difference was significant only for serum 2 samples (collected a median of 15.3 months before the diagnosis of lymphoma) (Table 2). EBV DNA was positive in PBMC samples from all tested cases during the 3 years preceding the diagnosis of cerebral lymphoma, but it was also positive in 87 to 94% of controls during the same period (Fig. 2a). EBV DNA was not more often detectable (> threshold or detectable < threshold) in sera from cases than in sera from controls (with 0 to 23.1% positive detection in the last 3 years for cases vs. 4.8 to 12% for controls) (Fig. 2b).

1b). A terminator was predicted by webgester downstream of yaaH or, in antisense at the same position, downstream of mog. This suggests that yaaW is most likely organized as operon yaaIWH in EHEC and transcribed from the yaaI-promoter and terminated downstream of yaaH.

Interestingly, data from genexpdb indicate that htgA and yaaW are expressed differentially in E. coli strains under certain experimental conditions (see Table 1), clearly prohibiting htgA synonymizing with yaaW, which has been performed in some databases. HtgA and YaaW were expressed in EDL933 using a plasmid that generates concomitant myc and His-tag fusions. Proteins were prepurified using the his-tag and detected on Western blots using the myc-tag. YaaW (30 kDa) was detectable, but no band for HtgA was found (Fig. 2), which is in accordance with Narra et al. Alpelisib nmr (2008). Thus, the protein might be unstable Selleck Gefitinib and difficult to discover. Missiakas et al. (1993) presented a 21-kDa gene product by 35S-labeling, which is a more sensitive approach. Previous work always used a double knockout mutant. We created strand-specific deletion mutants for the first time, in which only htgA or yaaW was interrupted (Fig. 3). The annotated htgA-start codon is CTG, which is quite rare for bacteria. The next GTG is more likely to be the start codon. Counting from there, htgA has 525 bp (or 174 amino acids); our htgA-knock out terminates either product. By

introducing a single-point mutation to create a stop in one frame, we minimized the disturbance of the other, as the mutations are synonymous in the latter (Tunca et al., 2009). For the first time, it was possible to distinguish

effects of ΔhtgA from ΔyaaW. Both mutants showed no difference in their growth compared with wild type at 37 °C or after temperature shift from 30 °C to 45 °C (Fig. 4a). As no heat shock phenotype of ΔhtgA could be confirmed (as found before, Nonaka et al., 2006), htgA should no longer be annotated as heat shock gene. In minimal medium, biofilm formation of ΔhtgA or ΔyaaW was reliably increased when incubated for 48 h at 37 °C (Fig. 4b). This is in accordance with Domka et al. (2007), who found a threefold increase in biofilm formation for E. coli K12 in a htgA/yaaW double mutant. We speculate ALOX15 that the higher increase compared with our experiments might be due to additive effects of both genes in the double mutant compared with each single one. We therefore suggest to rename htgA to mbiA (modifier of biofilm). As no difference in growth could be found, we measured the metabotypes. Metabolite changes could still be detectable even though they may not manifest in growth (Raamsdonk et al., 2001). ΔhtgA, ΔyaaW, and wild type were subjected to nontargeted metabolomics using ICR-FT/MS. Indeed, twenty-two different metabolites (putatively annotated, see Table S3) between the strains were found significantly changed (P ≤ 0.01).