We assessed the production of OMVs from K. pneumoniae ATCC 13883 during in vitro culture. Bacteria were cultured in LB broth, and OMVs were purified from culture supernatants. TEM analysis showed that K. pneumoniae-derived vesicles were spherical bilayered structures with diameters of 20–200 nm (Fig. 1a). No bacteria or protein contaminants were observed. The small-sized OMVs with diameters of approximately 20–30 nm were commonly observed, whereas relatively

large-sized vesicles with diameters of > 50 nm were less commonly observed. This result suggests that K. pneumoniae produces and secretes OMVs into the extracellular milieu during in vitro culture. Klebsiella pneumoniae OMVs were subjected to SDS-PAGE. Many protein bands were identified in the K. pneumoniae OMVs, but the protein profiles were different between OMVs and whole-cell lysates (Fig. 1b), DNA Synthesis inhibitor suggesting the absence of bacterial contaminants. Proteomic analysis was conducted to identify proteins in the OMVs from K. pneumoniae ATCC 13883. We identified

159 proteins in the K. pneumoniae OMVs (Supporting Information, Table S1). The proteins identified in the K. pneumoniae selleck compound OMVs were predicted to occur in the extracellular space (n = 13), outer membrane (n = 24), periplasmic space (n = 25), inner membrane (n = 13) and cytoplasm (n = 84). Of the proteins identified in the K. pneumoniae OMVs, the outer membrane protein X, murein lipoprotein, phage shock protein: Histone demethylase activates phage shock-protein expression, putative uncharacterized protein ygdR and 30S ribosomal protein S20 were the most abundant among the proteins located in the

outer membrane, periplasmic space, inner membrane, extracellular space and cytoplasm, respectively. These results suggest that K. pneumoniae OMVs contain numerous proteins originating from inner membrane and cytoplasm as well as outer membrane and periplasmic space. OMVs are naturally secreted products of Gram-negative bacteria, and OMV production appears to be a conserved process among Gram-negative bacteria (Beveridge, 1999; Kuehn & Kesty, 2005; Kulp & Kuehn, 2010). Additionally, Gram-positive bacteria such as Staphylococcus aureus and Bacillus anthracis also produce membrane-derived vesicles (Lee et al., 2009; Rivera et al., 2010; Gurung et al., 2011). Deatherage et al. (2009) proposed the OMV biogenesis model in Salmonella typhimurium. During bacterial growth and division, localized reductions in the density of outer membrane–peptidoglycan and outer membrane–peptidoglycan–inner membrane associations result in the bulging and release of the outer membrane as OMVs. Based on this model, OMVs only reflect the outer membrane and periplasmic components. However, cytoplasmic and inner membrane proteins have been identified in OMVs from E. coli (Lee et al., 2008), H. pylori (Olofsson et al., 2010) and Acinetobacter baumannii (Kwon et al., 2009).

The patient and her parents and sister were subjected to microbiological testing to identify the microorganisms involved in the disease. The patient underwent tooth extraction to eradicate the

disease and received a prosthesis for the restoration of masticatory function. After the permanent teeth erupted, fixed orthodontic appliances were place to restore dental arch form and occlusion. Conclusions.  The results show the importance of an early diagnosis of GAP and of a multidisciplinary selleck screening library approach involving laboratory and clinical management to treat the disease and to restore masticatory function, providing a better quality of life for patients. “
“International Journal of Paediatric Dentistry 2010; 20: 293–304 Background.  Existing indices to quantify tooth discolouration are mostly aetiology-specific. An index of tooth appearance (IOTA), derived from all types of tooth discolouration and surface defects, would allow the quantification of attractiveness for psychological assessment and treatment planning Objective.  To develop a perception based IOTA for quantification of all forms of tooth discolouration and surface defects. Methods.  One hundred images of discoloured teeth were twice ranked by a panel of judges according to perceived

attractiveness. Mean image score was then used to arrange the images into a continuum of attractiveness and from these, ten images were selected, to constitute the illustrated IOTA. A second panel of judges assessed 35 clinical pictures 3MA using the IOTA, on two occasions. Results.  The first 100 images were assessed with a correlation of 0.79–0.81 between the two ranking sessions and with intra-group reproducibility of 0.8–0.94. The second panel of judges used the developed IOTA quickly, with an intra-judge correlation of 0.87 and inter-judge reliability of 0.72 and 0.74 for two sessions. Conclusions.  The IOTA could be

used by clinicians as a tool for quantifying disfigurement in teeth, irrespective of aetiology or histology. “
“International Journal of Paediatric Dentistry 2011; 21: 1–12 Background.  Several tools have been developed for the measurement www.selleck.co.jp/products/sorafenib.html of emotional status of the child in paediatric dental clinics including nonverbal self-report techniques. Subjective methods like drawing and Child Drawing: Hospital (CD:H) score have recently been applied in hospitalized children. Studies, however, have not attempted to analyse children’s drawings as an aid to investigate the subjective feelings of children in paediatric dental settings. Aim.  To assess drawing as a measure for child’s distress in paediatric dental settings. Design.  Fifty-four children, aged 4–11 years, participated in this study. After finishing the first therapeutic session, the child was instructed to draw a picture of a person in a dental clinic. The pictures were scored using CD:H score sheet and the findings were compared with SEM and Frankl scores.

The views and conclusions contained hereon are those of the authors and should not be interpreted as necessarily representing

the official policies or endorsements, either expressed or MG-132 in vivo implied, of IARPA, DOI, or the US Government. Abbreviations ACh acetylcholine BF basal forebrain Glu glutamate LGN lateral geniculate nucleus mAChRs muscarinic acetylcholine receptors PFC prefrontal cortex TRN thalamic reticular nucleus V1 primary visual cortex “
“miR-96 is a microRNA, a non-coding RNA gene which regulates a wide array of downstream genes. The miR-96 mouse mutant diminuendo exhibits deafness and arrested hair cell functional and morphological differentiation. We have previously shown that several genes are markedly downregulated in the diminuendo organ of Corti; one of these is Ptprq, a gene known to be important for maturation and maintenance of hair cells.

In order to study the contribution that downregulation of Ptprq makes to the diminuendo phenotype, we carried out microarrays, scanning electron microscopy and single hair cell electrophysiology to compare diminuendo mutants (heterozygous and homozygous) with mice homozygous for a functional null allele of Ptprq. In terms of both morphology and electrophysiology, the auditory phenotype of mice lacking Ptprq resembles that of diminuendo heterozygotes, while diminuendo homozygotes are more severely affected. A comparison of transcriptomes indicates there is a broad similarity between diminuendo homozygotes Dapagliflozin and Ptprq-null mice. The reduction in Ptprq observed in diminuendo Bortezomib solubility dmso mice appears to be a major contributor to the morphological, transcriptional and electrophysiological phenotype, but does not account for the complete diminuendo phenotype. “
“The dopaminergic projections to the basal ganglia have long been implicated in reward-guided behavior and decision-making, yet little is known about the role of the posterior pedunculopontine nucleus (pPPN), a major source of excitatory input to the mesolimbic dopamine

system. Here we studied the contributions of the pPPN to decision-making under risk, using excitoxic lesions and reversible inactivation in rats. Rats could choose between two options – a small but certain reward on one lever; or a large but uncertain reward on the other lever. The overall payoff associated with each choice is the same, but the reward variance (risk) associated with the risky choice is much higher. In Experiment 1, we showed that excitotoxic lesions of the pPPN before training did not affect acquisition of lever pressing. But whereas the controls strongly preferred the safe choice, the lesioned rats did not. In Experiment 2, we found that muscimol inactivation of the pPPN also produced similar effects, but reversibly. These results show that permanent lesions or reversible inactivation of the pPPN both abolish risk aversion in decision-making.

e. peripheral

blood mononuclear cells, cerebrospinal fluid, lymph node tissue) [7,8]. As stated, one step toward improving our understanding of ARV pharmacokinetics is to measure FU in plasma as FU is free to traverse biological membranes, penetrate target tissue and exert pharmacological effect [9]. Other factors also contribute to how well a drug reaches its site of action, including the degree to which a drug is influenced by transport proteins (e.g. P-glycoprotein) present on cellular membranes [9]. Changes in PB will be most clinically relevant for highly bound drugs, since small changes in binding of highly bound drugs can have substantial effects on FU [10]. For highly bound ARV drugs such as the PIs, clinical situations likely to impact FU include pregnancy, infancy, renal or liver disease or concomitant therapies that displace drugs from PB sites [11–13]. This is one of the first studies addressing the effect GSK2126458 supplier of pregnancy on PB of any ARV drug despite long-term knowledge that pregnancy causes substantial reductions in plasma protein concentrations.

Albumin concentrations fall from 3.5 to a range of 2.5 to 3.0 mg/dL during the first half of pregnancy and steroid and placental LY2606368 nmr hormones displace drugs from binding sites leading to an overall decrease in the binding capacity of albumin and an increase in FU [14]. For example, the FU of phenytoin, a drug that binds to albumin, increases during pregnancy [14]. Data describing changes in AAG with pregnancy are less definitive. Two studies report an overall decrease in AAG concentration over the course of pregnancy while others report no change [11,12]. PB during pregnancy also may be affected by increased concentrations of endogenous ligands (i.e. free fatty acids), that may compete for drug binding sites of albumin and AAG. LPV has been reported to bind to both albumin and AAG, as does the PI nelfinavir [15,16]. AAG and albumin concentrations in our subjects were significantly altered during pregnancy compared to PP (P<0.0001). This is expected due to the change in weight and fluid volume consistent with pregnancy. The PAK5 weight gain in

our subjects of up to 10–15 kg during the course of pregnancy is also expected, although mean total weight for our subjects is higher than weights expected for pregnant women in some international settings where women on average are of smaller stature. Weight gain alone is not expected to impact FU. Albumin concentrations did not appear significantly to influence FU, but AAG concentrations had a significant effect, in that each 100 mg/L increase in AAG was associated with a decrease in LPV FU of 0.07 and 0.05% for third trimester and PP evaluations, respectively (P<0.0001 during both time periods, with adjustment for total LPV, at the PP evaluation). In contrast, LPV dose and time of PP evaluations did not have a significant effect on FU.

uberis based on colonial appearance, Gram stain reaction and catalase test (National Mastitis Council, 2004) and by conventional identification (Odierno et al., 2006). The selected colonies were maintained frozen at −20 °C in Todd–Hewitt broth (Sigma-Aldrich

Co.) containing 20% glycerol for further characterization. Isolates were identified as representing S. uberis by restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene according to Jayarao this website et al. (1992). All the isolates were additionally confirmed by RFLP analysis of the 16S rRNA gene, using the restriction enzymes RsaI and AvaII (Khan et al., 2003). Streptococcus uberis ATCC 27958 and Streptococcus parauberis ATCC 13386 were used as reference strains. The target genes, the oligonucleotide primers used and the sizes of the amplicons are summarized in Table 1. Synergistic CAMP-like haemolytic

activities were determined together with a β-toxin-producing Staphylococcus aureus on sheep blood agar plates (Odierno et al., 2006). Genomic DNA was isolated as described by Jayarao et al. (1992), purified by ethanol precipitation and dissolved in a buffer containing 10 mM Tris/HCl (pH 7.6) and 0.1 mM EDTA. Specific oligonucleotide primers for the detection of the cfu, lbp and sua genes of S. uberis were designed for this study with primer3 software (http://frodo.wi.mit.edu/primer3/). anti-PD-1 monoclonal antibody DNA amplification for the hasA, hasB, hasC, oppF, pauA/B and skc genes was performed using oligonucleotide primers derived from published sequences. All the oligonucleotides were synthesized by Promega

Corporation. The PCR was standardized for the detection of each virulence-associated gene following the methodologies described with suitable modifications to optimize the different conditions that affect the sensitivity and specificity of the reaction. Carnitine dehydrogenase Details of the primer sequences are shown in Table 1. To amplify the genes, 50 μL of reaction mixture was made containing 20 ng template DNA, 1 μM oligonucleotide primers, 0.4 μM of each of the four dNTPs, 1.50 U Taq polymerase and 1.5 mM MgCl2. The annealing temperature was varied from 48 to 58 °C depending on the gene being amplified. The reactions were carried out in a thermal cycler and genes of each isolate were tested at least twice. A positive and a negative control were included in each run. PCR products were resolved on 1.2% agarose gel at 70 V for 1.5 h. Gels were stained with ethidium bromide solution (0.5 mg mL−1) and photographed under UV light with MiniBisPRO gel documentation. RFLP analysis of the 16S rRNA gene successfully identified 78 isolates as S. uberis at the molecular level based on comparisons with reference strain S. uberis ATCC 27958 (Fig. 1). A synergistic haemolytic CAMP-like reaction on sheep blood agar within the zone of staphylococcal β-toxin could be observed for 18 of the 78 (23%) S. uberis strains. The standardized PCR allowed the amplification of putative and known virulence-associated genes of S.

The most common symptom at diagnosis was a seizure. The average interval between return from the suspected travel and symptom onset was 3.2 ± 1.8 years. Two patients suffered from multiple lesions, whereas the rest had a single lesion. Antihelminthic treatment was given to most patients with resolution of symptoms. Median duration of antiepileptic treatment was 16 ± 41 months after albendazole was given. Antiepileptic treatment LY2109761 was discontinued without any complications. The estimated attack rate of clinical disease was 1 : 275,000 per travel episode to an endemic region. Conclusions. NCC

in travelers is a rare phenomenon commonly presenting as seizure disorder manifesting months to years post-travel. Antihelminthic therapy followed by 12 to 24 months of antiepileptic therapy resulted in complete resolution of symptoms in our patients. Neurocysticercosis (NCC) is an infection of the central nervous system (CNS), caused by the larval stage of the pork tapeworm, Taenia

solium. NCC is considered the most common parasitic disease of the human nervous system.1,2 It is also the most common cause of acquired epilepsy in developing countries.3 The disease is common throughout Latin America, Asia, ABT-199 molecular weight sub-Saharan Africa, and parts of Oceania. In developed countries, NCC is usually encountered among immigrant populations from endemic areas.4 Humans are regarded as the only definitive hosts of T. solium, although it was recently reported that pigs may undergo secondary infection by primarily infected pigs.5 The causative agent, T. solium, has a distinctive life cycle, causing two distinct clinical syndromes in the human host. Ingestion of raw pork meat contaminated with T. solium larvae results in larval maturation into adult cestodes in the small intestine, causing human taeniasis (Figure 1a). Fecal excretion of gravid proglottids begins approximately 2 months after infection. The worm attaches to the small intestine mucosa causing

mild inflammation, which may result in such symptoms as abdominal discomfort, nausea, and diarrhea. However, the host is usually unaware of the infection or of the proglottids in the stools. The second clinical syndrome, human cysticercosis, is initiated by ingestion of T. solium ova, usually as a consequence of fecal–oral transmission. This can be either autoinfection, due Phospholipase D1 to poor hygiene and self-transmission by the hands of the human host, or heteroinfection may occur where food handlers are intestinal carriers of T. solium or where food and water carry fecal material.1 Once eggs are ingested, infective embryos hatch in the intestine, invade the intestinal wall, and migrate to striated muscles, as well as to the brain, liver, and other tissues, where they develop into cysticerci (Figure 1b). On reaching the target tissue, a cyst is formed. Outside the CNS, cysticercosis causes minor symptoms.6 However, the CNS is the main target in which the formation of cysts results in significant morbidity.

This was despite the overwhelming anatomical evidence for the many different subtypes of GABAergic inhibitory interneurones

that are found in the brain (e.g. Ramón y Cajal, 1909, 1911; Peters & Jones, 1984; Monyer & Markram, 2004; Butt et al., 2005; Klausberger & Somogyi, 2008). We now know a lot more about inhibitory circuitry and can begin to predict some of the many different roles that inhibitory connections play. In neocortex selleckchem alone there are > 20 different types of inhibitory connection in each of five layers (2–6). While it has been clear for a long time that gross alterations in inhibitory gain control result in coma on the one hand and seizures on the other, more recently, seemingly small or subtle changes in only one part of the inhibitory system have been linked to perhaps less life-threatening, but nevertheless Silmitasertib ic50 debilitating and tragic, neurological and psychiatric disease. Similarly, pharmacological or genetic manipulation

of single GABAA receptor (GABAAR) subunits produces rather less dramatic, but nevertheless profound, alterations in mood and behaviour (Möhler, 2006a,b; Möhler et al., 2002; Rudolph & Möhler, 2006, for reviews). One important consequence of this selectivity is that, in many cases, changing the activity of a given GABAAR subtype, either by gene mutation, or by pharmacological BCKDHA manipulation, modifies the outputs of a given class of presynaptic GABAergic neurone, rather than modifying the inputs to a given class of postsynaptic neurone. These manipulations can, therefore, indicate the role(s) that different classes of interneurones play. Synaptic GABAARs are typically pentomers

containing a γ2 subunit in addition to two β- and two α-subunits. A β-subunit (β1, β2 or β3) is almost obligatory for plasma membrane insertion. Multimers without β-subunits are often destroyed before leaving the endoplasmic reticulum (ER). β-Subunits may also play a role in subcellular compartment-targeting of receptors: to axon, soma or dendrites. The α-subunits appear to convey an even finer level of specificity, a striking example being the inputs provided by two major subclasses of basket cells to the soma of the same pyramidal cell in neocortex and hippocampus. The GABAARs innervated by parvalbumin (PV)-containing basket cells include α1-, β2/3- and γ2-subunits, while α2,β2/3,γ2-GABAARs are innervated by cholecystokinin (CCK) basket cells (Fig. 1; also Pawelzik et al., 1999; Thomson et al., 2000; Ali & Thomson, 2008). Benzodiazepines acting only at α1-GABAARs produce sedative and anticonvulsant effects, but generate anxiolysis only when they act at α2/3-GABAARs (Möhler et al., 2002), indicating the functional and potential therapeutic relevance of this differentiation.

It is difficult to tell what biological

processes exactly account for the observed abnormality of FA and MD values in patients with ADHD as the neuroanatomical GPCR Compound Library and physiological correlates of diffusion parameters are not yet entirely understood (Beaulieu, 2002; Versace et al., 2008). In our study, lower FA and higher MD in orbitofrontal areas of patients with ADHD may correlate with myelination deficits, changes in axonal integrity, lower packing density of fibres or more obliquely oriented fibres. However, higher FA and lower MD localized in frontotemporal WM, where fibres of several SCH772984 solubility dmso tracts (IFO, inferior longitudinal fasciculus, uncinate fasciculus) are

crossing (Mori et al., 2005), might rather be the result of less fibre crossings in this area. While higher FA in fibre tracts usually correlates with higher structural integrity, this correlation may not be correct in brain areas containing a particular high amount of fibre crossings, which results in lower FA values. In these areas, a higher number of fibres and thus a larger number of fibre crossings may result in higher connectivity of the involved brain areas and thus may lead to lower FA (Beaulieu, 2002). This may explain increased FA in patients with ADHD in frontotemporal WM clusters. In this context, it has

to be mentioned that age effects on FA and MD have been previously described in healthy adults (Sullivan & Pfefferbaum, 2006), although age effects are unlikely to account for the observed group differences in our study, because both groups did not differ significantly in age. Taken together and in light of previous neuroimaging studies, our finding SPTBN5 of orbitofrontal WM changes in adult patients with ADHD supports the notion of disturbed frontal-striatal circuitry in ADHD. DTI measures for WM integrity are in part directly correlated with impulsivity in this network, while attentional performance in patients with ADHD is correlated with microstructural properties in parts of the right SLF. Moreover, we provide further evidence for microstructural abnormalities in adult patients with ADHD in the cingulum bundle. Further studies combining refined functional and structural imaging methods are needed to investigate disturbed connectivity and their impact on behavioural measures in adult ADHD. We thank the patients and volunteers who participated in our study.

30) to form a single-beam optical trap. A R. glutinis cell in the phosphate-buffered

saline (PBS) was trapped about 10 μm above the bottom of cover slip with a gradient force generated by the focused beam. The same laser beam was used to excite Raman scattering from molecules inside the trapped cell. The spectrum was obtained by a liquid-nitrogen-cooled charge-coupled detector. The spectral resolution of our Raman system was about 6 cm−1. The Raman measurement of an individual cell was performed with a 10-s exposure time and 30 mW excitation power. The Raman spectra of 100 cells were collected for each time point. The PBS background spectrum was recorded with the same acquisition condition without the trapped cells and subtracted from the Inhibitor Library chemical structure spectra of individual cells. The subtracted spectra were then smoothed using the Adjacent-Averaging filter method. Preprocessing of spectral data was performed using matlab 7.0 software. The total carotenoid level in an individual cell was estimated from the peak intensity at 1509 cm−1 in its Raman spectrum. β-Carotene standard (purchased from Sigma-Aldrich) was dissolved in chloroform and diluted into a series of concentrations: 62.5, 125, 187.5, 250, 312.5, 375, 437.5, and 500 mg L−1. For each measurement, a 150-μL aliquot of β-carotene solution was added to the sealed holder and its

Raman spectrum was acquired with the same experimental parameters used for determining the cell spectra. The Raman spectrum of the pure chloroform was taken as background and subtracted from the above-mentioned spectra. A standard curve Ganetespib ic50 for carotenoid

quantification was linearly fitted by correlating the β-carotene concentration PRKACG with the peak intensity at 1518 cm−1 in its Raman spectrum. Carotenoids are a family of isoprenoids containing a characteristic polyene chain of conjugated double bonds. In R. glutinis cells, carotenoid pigments predominantly consist of β-carotene, torulene, and torularhodin (Sakaki et al., 2002). In this work, the Raman spectra of R. glutinis cells cultivated for 12 and 32 h, as well as the pure β-carotene standard were acquired in order to verify the existence of carotenoids in the investigated stain (Fig. 1). The three fundamental carotenoid bands at 1505–1520 cm−1 assigned to C=C (ν1) in-phase stretching, 1156 cm−1 assigned to C–C (ν2) stretching and 1005 cm−1 assigned to δ(C=CH) in-plane rocking modes of CH3 groups were clearly visible in all of the spectra. Thus, to a high degree of certainty, these peaks resulted from carotenoid compounds. The intensity of these peaks for R. glutinis cells cultivated for 32 h was more than 30 times higher than those for cells cultivated for just 12 h. It is noteworthy that the C=C (ν1) peak was at 1509 cm−1 for carotenoids present in cells, while it was at 1518 cm−1 for the β-carotene standard. This difference may be attributed to the fact that carotenoids usually bind to proteins or lipids in R.

parvula Te3T (=DSM 2008=ATCC 10790=JCM 12972) has been published recently (Gronow et al., 2010), which makes this species an attractive model for C646 order in-depth analysis of the biology and pathogenesis potential of veillonellae as a group. Another strain, V. parvula PK1910 [formerly Veillonella atypica PK1910 (Hughes et al., 1992), Veillonella spp. PK1910 (Periasamy & Kolenbrander, 2009)], has been the most characterized Veillonella strain in the oral biofilm. The genome of PK1910 was recently sequenced by our group. Analysis of the draft sequence (http://www.oralgen.lanl.gov/) identified many genes homologous to the competence related genes of both gram-positive and gram-negative bacteria

(Qi & Ferretti, 2011), suggesting that this strain might be transformable. The objective of this investigation was to test the transformability of V. parvula PK1910. Using spontaneous and PCR-generated mutations in the rpsL gene, which

confers streptinomycin-resistance, we demonstrated that DNA containing these mutations could be transferred into PK1910 via electroporation and integrated into the chromosome possibly through homologous recombination. To our knowledge, this is the first report of genetic transformation in veillonellae. The bacterial strains and plasmids used in this study are listed in Table 1. Veillonella parvula strain PK1910 was formerly named V. atypica PK1910 or Veillonella spp. PK1910 (Hughes et al., 1992; Periasamy & Kolenbrander, 2009) and is now renamed V. parvula PK1910 based on MLN0128 in vitro our recent sequence analysis using the rpoB gene (Qi & Ferretti, 2011). Veillonella parvula PK1910 was grown in Todd–Hewitt (TH) broth (Difco) supplemented with 0.6% sodium lactate (THL), or brain heart infusion (BHI) broth (Difco) supplemented with 0.6% sodium lactate (BHIL), or a chemically defined medium (He et al., 2008) without glucose but supplemented with 0.6% sodium lactate and 0.1% peptone (ASSPL). Streptomycin

(Sigma Chemical Co.) was added to the medium at a final concentration of Cell press 1 mg mL−1 for mutant selection. All V. parvula PK1910 cultures were grown anaerobically (85% nitrogen, 5% carbon dioxide, 10% hydrogen) at 37 °C. Escherichia coli cells were grown in Luria–Bertani (LB; Difco) broth with aeration at 37 °C. Escherichia coli strains carrying plasmid was grown in LB containing 100 μg mL−1 ampicillin (Fluka). Veillonella parvula PK1910 overnight culture was plated on THL plates supplemented with 1 mg mL−1 streptomycin and colonies grown on the plates were isolated and purified. Chromosomal DNA was isolated from these mutants, and then the rpsL gene fragment was generated by PCR using primers rpsL-F and rpsL-R (Table 2 and Fig. 1) and sequenced. Veillonella parvula PK1910 cells were grown in THL, BHIL, or ASSPL media to designated growth phases (OD600 nm of 0.15–0.6), and harvested by centrifugation.