, 2001), amino acid substitutions in WX IdpA should affect the properties of the WX cell surface and subsequently increase the evolutionary fitness of WX. These results suggest that IdpA has an important role in host–phytoplasma interactions, particularly in WX. PFT�� mw Further sequence comparisons of Imp or IdpA among several strains of WX would reveal the functional importance of Imps in 16SrIII ribosomal group phytoplasmas. Most of the 30 poinsettia cultivars examined in this study

were infected with PoiBI, as shown by PCR amplification of the phytoplasma 16S rRNA gene, but phytoplasma infection could not be detected in four of the cultivars: ‘Flaming Sphere’, ‘Annette Hegg Marble’, ‘Annette Hegg Diva’, and ‘Eckespoint C-1 Red’. ‘Eckespoint C-1 Red’ was previously reported to be phytoplasma-free, along with ‘Eckespoint C-1 White’ (Dole & Wilkins, 1991), in agreement with our results. However, we cannot exclude the possibility that PCR failures resulted in false negatives for some or all of these cultivars. PCR failures could have arisen if the level of PoiBI accumulation was very low, perhaps as a result of the particular cultivar characteristics, growth stage, or growth conditions, or if the cultivar(s) contained PCR inhibitory compounds. Alternatively, the possibility of the sequence variability in the PCR primer

binding region cannot be excluded. It is possible that nested-PCR using 16SrIII group-specific primers, instead of the single PCR using the primers in this study, might yield amplification products. However, we extracted the template DNA for all samples from the poinsettia leaf midribs, where the concentration of phytoplasma cells http://www.selleckchem.com/products/Roscovitine.html is

expected to be high, and we followed the same extraction Interleukin-2 receptor protocol for all poinsettia cultivars. To eliminate the influence of PCR inhibitory compounds, we used DNA diluted by tenth and hundredth as a template for PCR amplification. However, we could not yield fragments from all of four cultivars (data not shown). Moreover, phenotypically, these four cultivars are taller and had less branching than PoiBI-infected poinsettias (Fig. S2). These features were similar to those of the healthy poinsettia. Therefore, we conclude that in addition to ‘Eckespoint C-1 Red’ and ‘Eckespoint C-1 White’, several other commercially available poinsettias, that is, ‘Flaming Sphere’, ‘Annette Hegg Marble’, and ‘Annette Hegg Diva’, are free of phytoplasma infection. The conservation of Imp sequences among many groups of phytoplasmas has led to the suggestion that Imp represents an ancestral type of Imp (Kakizawa et al., 2009). This proposal suggests that PoiBI has retained Imp as its major membrane protein, and that the expression level of IdpA had increased during the evolution of WX, causing IdpA to become the Imp of WX. It is known that WX is transmitted predominantly by Colladonus montanus (Kirkpatrick et al., 1987), whereas it has been assumed that PoiBI is transmitted only by grafting.

, 2004; Christianson et al., 2010) or both structures (Wu et al., 1999; Yang et al., 2008), Lino-de-Oliveira et al. (2002, 2006) showed that microinjections of glutamate into the DPAG reduce floating behavior whereas those of lidocaine had the opposite effect. Most importantly, the latter authors also showed that sub-chronic administrations of antidepressants reduce FST-induced increases in fos-like immunoreactivity in most columns of the PAG (Lino-de-Oliveira et al., 2002, 2006). Most notably, TSA HDAC research buy however, recent data from positron-emission tomography

in rats (microPET) showed that whereas the PAG is markedly activated during FST training session, it remains inactive in test sessions (Jang et al., 2009). Lastly, plenty of evidence suggests that the protective effect of controllable stress is mediated by prefrontal cortex efferents to neurons of dorsal raphe (DR) which project to the DPAG (Maier CX5461 et al., 1995; Neumaier et al., 1997; Amat et al., 2005, 2006; Maier & Watkins, 2005; Rozeske et al., 2011). However, whereas the DR is the target of prefrontal cortex projections, predominantly inhibitory (Celada et al.,2001; Goncalves et al., 2009), the stimulation of DR either excites (directly) or inhibits (indirectly) the DPAG (Stezhka & Lovick, 1994). Moreover, DPAG levels of 5-HT did

not change either during exposure to IS (Amat et al., 1998) or 1 week thereafter (C.A. Rosa, unpublished results). Although the nature of DPAG-evoked freezing remains a matter of debate (De Oca et al., 1998; Schenberg et al., 2001, 2005; Vianna et al., 2001a,b), it is noteworthy that freezing was also markedly attenuated 1 week after exposure new to IS. Consequently, the attenuation of DPAG-evoked escape behaviors cannot be ascribed to a facilitation of freezing at the

expense of trotting and galloping behaviors. Alternatively, the impairment of both passive (freezing) and active (flight) behaviors is best explained by the inhibition of a DPAG in-built motivational system. Therefore, rather than a ‘panicolytic effect’, the attenuation of elevated T-maze (De Paula Soares et al., 2011) and DPAG-evoked escape behaviors following the exposure to uncontrollable stress may be a reflection of a decrease in resilience to stress. This possibility is supported by the much greater attenuation of trotting and galloping, the responses most effective in one-way shuttle-box escape training, than of jumping. Strong et al. (2011) suggested, on the other hand, that 5-HT2C receptors of dorsal striatum (DS) play a crucial role in learning deficits of inescapably-shocked rats. In particular, whereas the microinjections of a 5-HT2C receptor antagonist into DS prevented the escape failure, microinjections of a respective agonist impaired learning even in the absence of prior exposure to IS. Accordingly, it is tempting to speculate that DPAG and DS mediate, respectively, motivational and learning aspects of helplessness.

4b and c). These results suggest that A. actinomycetemcomitans-expressed leukotoxin induced the release Volasertib price of resistin by degranulation of the neutrophils before cytolysis. To examine the possible involvement of LFA-1, which is the receptor for leukotoxin, and an Src family

tyrosine kinase in the release of resistin and elastase, we pretreated neutrophils with monoclonal TS1/22 (CD11a) or TS1/18 (CD18) antibody against LFA-1 subunits at a final concentration of 5 μg mL−1 and with 10 μM PP1 for 15 min, followed by incubation with wild-type or mutant HK921 for 2 h (Fig. 5a and b). In contrast to pretreatment with TS1/22, the level of resistin released from neutrophils pretreated with TS1/18 or PP1 was significantly lower than that from untreated neutrophils after incubation with wild-type HK921,

as was the release of elastase. Additionally, the inhibitory effect of pretreatment with TS1/18 or PP1 on the levels of resistin and elastase released from neutrophils after incubation with mutant HK921 for 2 h was lower than those with wild-type HK921, but significant (P<0.05) (Fig. 5a and b). Among the several virulence factors expressed by A. actinomycetemcomitans, leukotoxin is thought to play a major role in disease progression (Henderson et al., 2003). Leukotoxin has been reported to activate cytolysis of human leukocytes, including neutrophils and monocytes CX-5461 manufacturer (Taichman et al., 1980), and to induce caspase-1 activation and bioactive IL-1β secretion in human macrophages (Kelk et al., 2003). In the present study,

significantly more resistin was released from neutrophils incubated with wild-type HK921, which is characterized by a 530-bp deletion of the promoter region of the leukotoxin gene, than from neutrophils incubated with minimally leukotoxic strains. The ability of the wild-type strain to produce leukotoxin was confirmed by Western blot analysis using a leukotoxin-specific antiserum, and its cytotoxic activity against neutrophils was demonstrated by LDH release. Furthermore, the mutant strain, medroxyprogesterone which is incapable of producing leukotoxin, released a significantly lower level of resistin (P<0.05). Whereas resistin is derived exclusively from adipose tissue in mice, leukocytes are the major source of resistin in humans. Neutrophils store abundant amounts of resistin in their granules, and the extracellular release of resistin via degranulation may be stimulated by bacterial or inflammatory stimuli (Bostrom et al., 2009; Johansson et al., 2009; Kunnari et al., 2009). This study demonstrated that the release of resistin from neutrophils in the presence of a highly leukotoxic strain, which is strongly associated with aggressive periodontitis in certain susceptible human populations (Haubek et al., 2008), was significantly higher than in the presence of a leukotoxin-deficient isogenic mutant (P<0.05).

baumannii BM4547 and P. aeruginosa PU21

as recipients and the five NDM-1-positive E. coli J53 transconjugants as donors. Mixes of donor and recipients cells were incubated for 18 h at 37 °C for S. typhimurium LT2, A. baumannii BM4547, P. aeruginosa PU21 and P. mirabilis CIP103181 and for 3 h at 37 °C for K. pneumoniae CIP15153. In addition, E. coli J53 transconjugant carrying a c. 70-kb IncF-type blaCTX-M-15-positive plasmid was included for comparison, as IncF-type plasmids conjugate efficiently among Enterobacteriaceae (personal data). Transfer frequencies were calculated by dividing the number of transconjugants by the number of donor cells. Statistical analysis was performed PS 341 using the Student’s t-test; a P-value of ≤ 0.05 was considered significant. Transformation experiments were performed as described previously by electroporation Selleckchem Target Selective Inhibitor Library of a plasmid DNA suspension from the five NDM-1-positive E. coli J53 into

rifampicin-resistant P. aeruginosa and A. baumannii reference strains (Potron et al., 2009). pAT-RTG-4 (shuttle vector) and pInt-Veb plasmids were used as positive control for electroporation in A. baumannii and P. aeruginosa (Aubert et al., 2003; Potron et al., 2009). Selection was performed on agar plates supplemented with ticarcillin (50 μg mL−1). MICs of carbapenems and cefotaxime were determined using the E-test strips (bioMérieux, Marcy l’Etoile, France). The five blaNDM-1-carrying plasmids of Enterobacteriaceae studied here belonged to various incompatibility groups (L/M, FII, A/C and two untypeable plasmids). IncL/M, IncA/C and IncFII plasmid types have been frequently described in Enterobacteriaceae carrying other β-lactam resistance determinants (Carattoli, 2009). IncL/M- and IncA/C-type plasmids

are broad-host range plasmids, whereas IncF-type plasmids are narrow-host range plasmids (Novais et al., 2007). learn more The five NDM-1-positive plasmids were self-conjugative using E. coli J53 as recipient at frequencies ranging from 10−4 to 10−8 transconjugants/donor (Table 1). The blaNDM-1 gene was the single carbapenem resistance marker located on those plasmids. Using blaNDM-1-positive E. coli J53 transconjugants as donors, second-step transconjugants were obtained using E. coli, K. pneumoniae, S. typhimurium and P. mirabilis as recipient species. In E. coli JM109, transfer frequencies ranged from 10−4 to 10−8 transconjugants per donor depending on plasmid type (Table 2). The lowest transfer frequencies were obtained with the untypeable plasmid p419 and IncA/C-type plasmid pKp7. No difference of transfer rate was observed using E. coli Tc601 and E. coli Tc271 as donors when different temperatures were used during the mating-out assays (Table 2). Using Tc419 and TcKp7 as donors, the transfer rate was significantly higher at 30 °C compared with that observed at 25 °C and 37 °C (P < 0.05), as reported for other blaNDM-1-positive plasmids (Walsh et al., 2011). For E.

baumannii BM4547 and P. aeruginosa PU21

as recipients and the five NDM-1-positive E. coli J53 transconjugants as donors. Mixes of donor and recipients cells were incubated for 18 h at 37 °C for S. typhimurium LT2, A. baumannii BM4547, P. aeruginosa PU21 and P. mirabilis CIP103181 and for 3 h at 37 °C for K. pneumoniae CIP15153. In addition, E. coli J53 transconjugant carrying a c. 70-kb IncF-type blaCTX-M-15-positive plasmid was included for comparison, as IncF-type plasmids conjugate efficiently among Enterobacteriaceae (personal data). Transfer frequencies were calculated by dividing the number of transconjugants by the number of donor cells. Statistical analysis was performed Gamma-secretase inhibitor using the Student’s t-test; a P-value of ≤ 0.05 was considered significant. Transformation experiments were performed as described previously by electroporation CAL-101 mw of a plasmid DNA suspension from the five NDM-1-positive E. coli J53 into

rifampicin-resistant P. aeruginosa and A. baumannii reference strains (Potron et al., 2009). pAT-RTG-4 (shuttle vector) and pInt-Veb plasmids were used as positive control for electroporation in A. baumannii and P. aeruginosa (Aubert et al., 2003; Potron et al., 2009). Selection was performed on agar plates supplemented with ticarcillin (50 μg mL−1). MICs of carbapenems and cefotaxime were determined using the E-test strips (bioMérieux, Marcy l’Etoile, France). The five blaNDM-1-carrying plasmids of Enterobacteriaceae studied here belonged to various incompatibility groups (L/M, FII, A/C and two untypeable plasmids). IncL/M, IncA/C and IncFII plasmid types have been frequently described in Enterobacteriaceae carrying other β-lactam resistance determinants (Carattoli, 2009). IncL/M- and IncA/C-type plasmids

are broad-host range plasmids, whereas IncF-type plasmids are narrow-host range plasmids (Novais et al., 2007). Astemizole The five NDM-1-positive plasmids were self-conjugative using E. coli J53 as recipient at frequencies ranging from 10−4 to 10−8 transconjugants/donor (Table 1). The blaNDM-1 gene was the single carbapenem resistance marker located on those plasmids. Using blaNDM-1-positive E. coli J53 transconjugants as donors, second-step transconjugants were obtained using E. coli, K. pneumoniae, S. typhimurium and P. mirabilis as recipient species. In E. coli JM109, transfer frequencies ranged from 10−4 to 10−8 transconjugants per donor depending on plasmid type (Table 2). The lowest transfer frequencies were obtained with the untypeable plasmid p419 and IncA/C-type plasmid pKp7. No difference of transfer rate was observed using E. coli Tc601 and E. coli Tc271 as donors when different temperatures were used during the mating-out assays (Table 2). Using Tc419 and TcKp7 as donors, the transfer rate was significantly higher at 30 °C compared with that observed at 25 °C and 37 °C (P < 0.05), as reported for other blaNDM-1-positive plasmids (Walsh et al., 2011). For E.

48, P = 0.03, Bonferroni corrected). Analysis of ipsilateral electrodes showed no P100 attention effect. A correlation of the ERP attention modulation and behavioural effect showed no significant relationship (r = 0.25, n.s). Analysis of the endogenous counter-predictive task showed no significant effects involving the factor Cue. There was a Task

× Cue × Hemisphere interaction (F2,22 = 7.05, P = 0.004,  = 0.39), as well as a main effect of Cue (F1,11 = 20.87, P = 0.001,  = 0.66) and Cue × Hemisphere interaction (F1,11 = 16.27, P = 0.002,  = 0.60). The significant interaction was further broken down into separate analysis for each task. Exogenous task analysis of the N140 showed a significant Cue × Electrode site × Hemisphere Alectinib in vivo interaction (F5,55 = 3.34, P = 0.029, http://www.selleckchem.com/products/VX-809.html  = 0.23), which was broken down into separate analyses for each hemisphere. However, there were no significant effects including the factor Cue at electrodes ipsilateral

or contralateral to the target presentation, indicating no attention modulation at the N140 in the exogenous task. Analysis of the endogenous predictive task revealed a significant main effect of Cue (F1,11 = 16.95, P = 0.002,  = 0.61), and also Cue × Hemisphere interaction (F1,11 = 21.53, P = 0.001,  = 0.66). The interaction was broken down revealing a significant effect of Cue, both for ipsilateral (F1,11 = 26.66, P < 0.001,  = 0.71) and contralateral

electrodes (F1,11 = 8.77, P = 0.013,  = 0.44), and both these effects showed enhanced negativity for expected compared with unexpected trials (the interaction was driven by larger effect size over ipsilateral compared with contralateral hemisphere; Fig. 4). That is, the N140 attention effect in the endogenous predictive task was present over both hemispheres. Moreover, and importantly, there was a significant correlation between the ERP attention modulation and the behavioural RT effect, with larger amplitude difference between expected Vildagliptin and unexpected conditions for each participant relating to larger RT attention effect (r = 0.69, P = 0.013; see Fig. 7 for a scatterplot of this relationship). The endogenous counter-predictive task revealed the attention effect was, similar to the endogenous predictive task, bilateral as there was a significant effect of Cue (F1,11 = 5.16, P = 0.044,  = 0.32). There was no significant correlation between ERP attention modulation and RT effect (r = 0.32, n.s.). At this last analysed time window the overall task analysis demonstrated a Task × Cue × Hemisphere interaction (F2,22 = 8.29, P = 0.002,  = 0.43) and also Cue (F1,11 = 11.02, P = 0.007,  = 0.50), and subsequently each task was analysed separately. The exogenous task revealed a Cue × Hemisphere interaction (F1,11 = 8.57, P = 0.014,  = 0.44).

Clinical examination revealed grade III mobile 71 and 81, with minimal

gingival inflammation and plaque deposits. There were no other dental findings and no significant medical history. Tooth numbers 71 and 81 exfoliated prematurely with no evidence of root resorption, shortly after presentation. Everolimus clinical trial Haematological and urinary investigations showed no abnormalities. Histological examination showed a complete absence of cementum. A diagnosis of OHP was made. After 10 months of dental follow-up, no further teeth have increased mobility. Conclusion.  Odontohypophosphatasia should be included as a differential diagnosis in children presenting with early loss of primary teeth. The dentist may be the first health care professional to whom the patient presents. “
“International Journal of Paediatric Dentistry 2013; 23: 32–38 Background  Salivary levels of Bifidobacteria have been shown to be significantly correlated with caries experience in adults but not as yet in children. Hypothesis.  Salivary levels of Bifidobacteria are

positively associated with caries experience in children. Aim.  To compare the salivary concentrations of Bifidobacteria of caries-free and caries-active children. Design.  Saliva was collected using the tongue-loop method from 38 caries-active children and from 22 clinically caries-free children, and the numbers of Bifidobacteria, mutans streptococci, lactobacilli and yeasts were determined. Additionally, the age and gender of the children, a plaque index, sugar amount in diet, sugar frequency in diet, hygiene practice and fluoride toothpaste usage were recorded. Results.  Bifidobacteria C59 wnt mw were isolated from 95% of the caries-active children and from only 9% of the caries-free children (P < 0.001). Salivary levels of Bifidobacteria Selleckchem Pembrolizumab were significantly correlated with amount of sugar in the diet, frequency of sugar consumption and oral hygiene practice. The significant variables that discriminated between the caries-free and caries-active subjects were salivary levels of Bifidobacteria, salivary levels of mutans streptococci

and oral hygiene practice (χ2 = 72.57, P < 0.001) and overall 90.0% of cases were correctly classified. Conclusions.  Salivary levels of Bifidobacteria are significantly associated with caries experience in children. The salivary levels of this genus may be a useful marker of caries risk. "
“This study aims to identify the determinants of caries prevention-oriented practice for children among final-year dental students in Nigeria. A questionnaire was distributed to 179 final-year dental students in six dental schools in Nigeria. It requested information on age, gender, knowledge of caries prevention measures, self-perceived competency in providing caries-preventive care for children, and caries prevention-oriented practice for two hypothetical cases with high and low risk of caries.

In ART-experienced patients who are virologically suppressed with an undetectable plasma HIV RNA level

(<50 copies/mL), the risk of hypersensitivity and/or hepatotoxicity on switching INCB018424 concentration to NVP is not increased in patients with higher CD4 cell counts (above the gender-specific CD4 cell count thresholds) [59]. In ART-experienced patients with detectable plasma HIV RNA levels, a switch to NVP is not advised. Furthermore, the need to minimize any window for developing resistance is greatest in patients who discontinue EFV early on when virological suppression has not yet been achieved. The latter scenario is made more complex when enzyme induction has not yet been fully achieved, and if doubt exists, alternatives to switch to should be considered. Steady-state (14 days following the

switch) ETV pharmacokinetic parameters are lowered by previous EFV intake in the case of both once-daily (Cmin was lowered by 33%) and twice-daily (Cmin was lowered by 37%) administration. However, ETV concentrations have been shown to increase over time following the switch and in patients with undetectable VLs switching from EFV to ETV, standard doses of ETV can be commenced [60]. To date, no data are available Selleck Ribociclib on what strategy to adopt in patients with active viral replication. Concentrations of RPV are lowered by previous EFV administration. However, 28 days after the switch, they returned to levels comparable with those when RPV was administered without previous EFV treatment, Cepharanthine except for a 25% lower Cmin.

Therefore, in patients with undetectable VLs switching from EFV to RPV, standard doses of RPV can be commenced [61]. To date, no data are available on what strategy to adopt in patients with active viral replication. Because of the strong inhibitory effect of ritonavir on CYP450 3A4, it is unlikely to require a modification of the PI/r dose when switching from EFV to PI/r. Formal pharmacokinetic data are unavailable. TDM data were presented on ATV/r and showed that after stopping EFV, ATV concentrations were above the suggested minimum effective concentration in all studied subjects [62]. Although formal pharmacokinetic data are not available, switching EFV to RAL should not lead to clinically significant consequences, as co-administration of EFV with RAL led to a moderate-to-weak reduction in RAL Cmin (21%) [63], which may persist for 2–4 weeks, after the switch but the degree of this reduction is unlikely to be clinically meaningful. A formal pharmacokinetic study in HIV-positive individuals showed that the induction effect of EFV necessitated an increase in MVC dose to 600 mg twice daily for 1 week following the switch [64]. MVC 300 mg twice daily (standard dose) seems to be safe after this period.

, 2009): selleck chemicals llc MD was significantly higher in patients with ADHD in these regions, but no difference was observed for FA values (Pavuluri et al., 2009). Moreover, decreased FA in the SLF and in the corticospinal tract in children and adolescents with ADHD has been demonstrated (Hamilton et al., 2008). Our

findings of increased FA bilaterally in frontotemporal WM connections point to an involvement of widespread brain areas in the pathophysiology of ADHD. While temporal structural abnormalities have not yet been described in previous MRI and DTI studies, a recent functional MRI study demonstrated bilateral temporal lobe dysfunction in boys with ADHD (Rubia et al., 2007). Possible reasons for the discrepancy with respect to the results of DTI studies in childhood and adolescence could be the sample heterogeneity between studies, the medication status of the investigated patients and the different diffusion imaging parameters between studies. In contrast to the majority of imaging studies in ADHD, we only included never-medicated patients in our study. Particularly, none of the patients had received any ADHD-specific treatment before such as psychostimulant medication. We are therefore able to exclude potential medication effects on imaging results as well as on neuropsychological findings.

In addition, we have excluded patients with ADHD with acute psychiatric comorbidity. Although Etoposide price we did not include medicated patients and patients with acute psychiatric comorbidity, symptom

severeness of our patients as measured with the BADDS was quite high (Brown, 1996; Table 1). For completeness, it also needs to be mentioned Reverse transcriptase that the possibility of false positive results in our study cannot be entirely excluded. In fact, taking into account the relatively weak group differences in our study, which would not survive a correction for multiple comparisons, and also the findings in DTI studies conducted by other groups in (adult) ADHD (Casey et al., 2007; Makris et al., 2008), replication studies would be desirable to confirm these findings. To our knowledge, this is the first study demonstrating a direct association between microstructural integrity and measures of attention in adult patients with ADHD. The correlation analyses between diffusion parameters and the ADHD score, which reflects the ability to focus attention (Greenberg & Kindschi, 1996), demonstrated significant findings in the right SLF. This specific fibre pathway together with the cingulum bundle connects frontal areas and cortical regions at the right temporo-occipito-parietal junction, which are considered to play a key role in processing information related to attentional functions (Makris et al., 2008).

Cat. Cmpd) including ETBR (Son et al., 2003). Overexpression of the Enterobacter cloacae sugE homolog

in E. coli generated cells with increased resistance to several QACs and ETBR (He et al., 2011). Overexpression of the Aeromonas molluscorum sugE homolog in E. coli generated resistance to ETBR, but not the QAC cetylpyridinium SB431542 chloride (Cruz et al., 2013). Also, when the E. coli SugE protein was assembled in membrane mimics, it bound to ETBR with a Kd in the low micromolar range, which is consistent with a role in ETBR transport (Sikora & Turner, 2005). Thus, in this work, we directly tested the model that Dcm influences sugE expression and thereby affects SugE-mediated resistance to antibacterial compounds. The bacterial strains used in this study are shown in Table 1 (Baba et al., 2006; Militello et al., 2012); plasmids were a gift from Ashok PD-1/PD-L1 tumor Bhagwat (Sohail et al., 1990). The lack of 5-methylcytosine in the dcm knockout strain JW1944-2 has been previously reported (Militello et al., 2012). The absence of the sugE gene in JW5738-1 and the rpoS gene in JW5437-1 was confirmed by PCR (data not shown). Liquid bacterial cultures were grown at 37 °C at 250 r.p.m. in either Luria Broth (LB) or M9 minimal media containing 0.4% glucose (Difco). Ampicillin was added to liquid cultures containing

dcm plasmids at 25 μg mL−1. Solid cultures were grown at 37 °C in the same media containing 15 grams of agar per liter, and when necessary ampicillin was added at 50 μg mL−1. All experiments to assess the sensitivity of strains to antibacterial compounds were performed in minimal media containing glucose as many QACs precipitate in LB. Bacteria were grown in LB at 37 °C at 250 r.p.m. to early logarithmic phase (A600 nm of c. 0.45) and early stationary phase (A600 nm c. 3.0). Total RNA was isolated from

3–4 mL of bacteria cultures using the MasterPure RNA Isolation oxyclozanide Kit (Epicentre). For 5-azacytidine experiments, the drug (Sigma-Aldrich) was dissolved in 1X phosphate-buffered saline (PBS), and PBS was added to untreated samples as a control. RNA quality was assessed using bioanalysis at the University of Rochester Genomics Research Center. Prior to reverse transcription, RNA was treated with RQ1 RNase-free DNase (Promega). One microgram of total RNA was used for reverse transcription using the New England BioLabs Protoscript kit with random primers. cDNA was used as a template for qPCR reactions on a Stratagene MX3000p machine. All reactions were run in triplicate or quadruplicate (technical replicates), and each experiment was performed 3–4 times (biological replicates). Data were normalized to the levels of malate dehydrogenase (mdh) using the ΔΔCt method (Livak & Schmittgen, 2001). The primer sequences are listed in Table S1.