Structures upstream tyrS represent the stems I, II, III and termi

Structures upstream tyrS represent the stems I, II, III and terminator of the leader region. The terminator/antiterminator

mechanism that regulates the tyrS gene is also indicated: readthrough of the leader region is induced by limitation of tyrosine. Uncharged tyrtRNA stabilize formation of SB202190 antiterminator structure in the mRNA, which prevents terminator formation (SD: Shine-Dalgarno; ORF: open reading frame of tyrS) Computational three-dimensional modelling of E. durans TyrS protein revealed nucleic acids-binding domains that might suggest a role as transcriptional regulator. However, the same domains have been identified in the highly similar TyrS structure of Thermus thermophilus (Protein Data Bank: 1H3E), and predicted to interact with tRNA (Figure 6). This data is consistent with the electrophoretic mobility shift (EMSA) assays carried to test TyrS binding to selleckchem the promoters of the TDC operon. Under the wide range of conditions studied (different pH, salt concentration, presence or absence of tyrosine…) no specific binding of TyrS was observed (data not shown). These data, together with the finding of tyramine clusters without a tyrS gene in Tetragenococcus halophilus

(GenBank AB059363) and histamine biosynthesis clusters without a hisS gene [36], would suggest a non critical biological function of these genes in the modulation of the contiguous decarboxylation operon. In any case, it can not be discarded that tyrS could exert a post-transcriptional regulation of tyramine biosynthesis. In fact, both enzymes -TyrS Mdivi1 datasheet and TdcA- share tyrosine as substrate. Figure 6 TyrS structural model achieved using Swiss-Pdb Viewer v. 4.04 software and structure superposition onto the highly similar Thermus terhmophilus tyrosyl-tRNA synthetase. (Protein Data Bank: 1H3E). 1H3E is shown in green, and TyrS model is shown in magenta and yellow. Protein kinase N1 Analysis of the two aligned structures indicates that all of the DNA/RNA binding

sites are in regions that interact with tRNA in the 1H3E structure (shown in blue). Consequently, two are the possibilities that can be considered: i) there are two tyrS genes in E. durans -as described for E. faecalis- and the one ligated to TDC would be a stress-related gene to ensure sufficient charged TyrtRNA for protein biosynthesis in those conditions that tyrosine is being decarboxylated, or ii) this is the unique tyrS gene and the low expression levels observed under neutral pH conditions are enough to assure protein synthesis for general metabolism and the increased expression at acidic pH would guarantee protein biosynthesis when tyrosine is being decarboxylated. The presence of a second tyrS gene was investigated by Southern hybridizations of E.

05, ANOVA, comparison for all pairs using Tukey test) IPS — Iod

05, ANOVA, comparison for all pairs using Tukey test). IPS — Iodophilic intracellular polysaccharides * MFar125F – myricetin, tt-farnesol and 125 ppm F; MFar250F – myricetin, tt-farnesol and 250 ppm F; 250F – 250 ppm F; Vehicle control – 20% ethanol containing 2.5% DMSO (v/v). ** Expressed as μg of phosphate released/mg of protein Figure 4 Influence of treatments on the pH values in the culture IBET762 medium during S. mutans biofilm formation. The medium was replaced daily with fresh medium. The pH values (n = 9) were determined

at 0 h and after 4, 8, 10 and 24 h of incubation each day. Values from vehicle control are significantly different from MFar250 at 10 h and 24 h of incubation, and from all treatments at 24 h of incubation during the entire experimental period (P < 0.05, ANOVA, comparison for all pairs using Tukey's test). Discussion Development of

novel chemotherapeutic approaches, other than microbiocides, that disrupt the establishment, structure and virulence of dental biofilms could be a promising route to prevent or reduce the learn more pathogenesis of oral infectious diseases such as dental caries. Currently, fluoride in various preparations is the mainstay for caries prevention [31]. Fluoride exerts its major effects by reducing enamel-dentine demineralization and enhancing remineralization of early caries lesions [18]. However, fluoride, at levels found in plaque, also displays biological effects on critical virulence factors of cariogenic streptococci, particularly (albeit not this website next exclusively) on S. mutans [10]. Nevertheless, as currently used, fluoride offers incomplete protection against dental caries (18). Thus, any agent that enhances its protective effects clearly has clinical potential. Recently, we have identified specific flavonoids (myricetin) and terpenoids (tt-farnesol) that exhibit bioactivity against S. mutans; these compounds are ubiquitously found in fruits (cranberries and red wine grapes) and propolis (a beehive product) [12, 13, 19, 20]. The concentrations of 1.0 mM myricetin and 2.5 mM tt-farnesol displayed the most potent inhibitory effects

on glucans synthesis and acid production by S. mutans cells as determined from our published and unpublished response to dose studies [13, 19, 20]. Furthermore, the combination of the naturally occurring agents with 250 ppm fluoride was the most effective in reducing S. mutans biofilm formation and EPS synthesis in vitro, and also enhanced cariostatic properties of fluoride in vivo [12, 13]. Analysis of our data shows that the natural agents acting in concert with fluoride (at 125 or 250 ppm) modulated the expression of specific virulence genes by S. mutans, and also disrupted the accumulation and structural organization of extracellular polysaccharides (EPS) and bacterial cells in the matrix, which affected the biochemical and physiological properties of the biofilms in vitro.

Adv Mater 2009, 21:2889 CrossRef 28 Zou RJ, Yu L, Zhang ZY, Chen

Adv Mater 2009, 21:2889.CrossRef 28. Zou RJ, Yu L, Zhang ZY, Chen ZG, Hu JQ: High-precision, large-domain three-dimensional manipulation of nano-materials for fabrication nanodevices. Nanoscale Res Lett 2011, 6:473.CrossRef 29. Zou RJ, Zhang ZY, Tian QW, Ma GX, Song GS, Chen ZG, Hu JQ: A mobile Sn nanowire inside a β‐Ga2O3 tube: a practical nanoscale electrically/thermally

driven switch. Small 2011, 7:3377.CrossRef 30. Splendiani A, Sun L, Zhang YB, Li TS, Kim J, Chim CY, Galli G, Wang F: Emerging photoluminescence in monolayer MoS2. Nano Lett 2010, 10:1271.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions YX carried out the this website exfoliation and fluorination and drafted the manuscript. QL, GH, KX, LJ, and XH participated in discussion of the study. YX and JH participated in the design

of the study and performed the statistical analysis. YX and JH conceived of the study, and participated in its design and coordination. All authors read and approved the final manuscript.”
“Background Electrical switching in the electrode/oxide/electrode structure has attracted significant attention due to its rich physics and potential application in the next generation nonvolatile memory [1]. A large variety of materials (such as metal CH5183284 cell line oxides, solid electrolytes, and organic materials) have been found to possess the characteristics of electrical switching [2–9]. Different models have also been proposed to understand the underlying physics of electrical switching [10–13]. However, the microscopic nature of https://www.selleckchem.com/products/MLN-2238.html electrical switching is still under debate, and exploring appropriate materials

for fabricating two-terminal resistive random access memory (RRAM) based on electrical switching is still the most important issue. Recently, nanoscale Pt/TiO2/Pt switches have been fabricated and well understood by memristive switching mechanism, in which Terminal deoxynucleotidyl transferase the drift of +2-charged oxygen vacancies under an applied electric field creates or annihilates conducting channels and then switches the device to on or off state [14, 15]. Therefore, nonstoichiometic oxides, in which oxygen vacancies play an important role on their electronic structures, might be the most appropriate materials for fabricating next generation nanoelectronic devices. Tungsten trioxide (WO3) has been investigated intensively because of its intriguing structural, electronic, and chromic properties [16–19]. Stoichiometic WO3 is resistive and transparent in the visible light region owing to a large band gap of 2.5 to 3.5 eV [16]. A slight deficit of oxygen (WO3−x , x = 1/6) is more favorable energetically than stoichiometic WO3 under atmospheric conditions, which implies that WO3 is intrinsically ‘self-doped’ by native oxygen vacancy point defects [17].

Louis, MO) Bacterial

Louis, MO). Bacterial Akt inhibitor strains L. pneumophila serogroup 1 strain AA100jm [39] is a spontaneous streptomycin-resistant mutant of strain 130b, which is virulent in guinea pigs, macrophages, and amoebae. The avirulent

dotO mutant was constructed by random transposon mutagenesis, as described previously [39]. This mutation results in severe defects in intracellular growth and evasion of the endocytic pathway [40]. The Corby flaA mutant derived from the wild-type Corby is defective in flagellin [41]. L. pneumophila strains were grown at 35°C in a humidified incubator on either buffered charcoal-yeast extract-agar medium supplemented with α-ketoglutarate (BCYE-α) or in buffered yeast extract broth supplemented with α-ketoglutarate (BYE-α). The flaA mutant was grown in an environment similar to those used for other Tipifarnib research buy strains, but in the presence of 20 μg/ml kanamycin. Heat-killed bacteria were prepared by heating the bacterial suspension at 56°C for 30 min or at 100°C for 1 h. Bacterial inactivation was achieved by treatment with paraformaldehyde (4%, 15 min followed by three washes in phosphate-buffered saline; PBS). Both types of treated suspensions were confirmed to contain no viable bacteria by plating them on BCYE-α agar. Cell culture Human T cells (Jurkat) were

maintained in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 100 U/ml PLX4032 penicillin G, and 100 μg/ml streptomycin. Human peripheral blood mononuclear cells (PBMC) were Phosphoprotein phosphatase isolated from peripheral blood of healthy donors using Ficoll-Hypaque gradients. PBMC were then further purified using positive selection with immunomagnetic beads specific for CD4 (Miltenyi Biotec, Auburn,

CA). On the day of the experiment, cells were refed with fresh antibiotic-free medium and cocultured with L. pneumophila for the time intervals indicated below. Infection of T cells and intracellular growth kinetics experiments Jurkat or CD4+ T cells seeded in plates were inoculated with either AA100jm or dotO mutant and either Corby or flaA mutant at an MOI of 100. In some experiments, heat-killed or paraformaldehyde-fixed bacteria were inoculated in the same manner. At 2 h after infection, cells were centrifuged and the supernatant was discarded. Cells were washed three times with PBS and resuspended in fresh RPMI 1640 medium containing 100 μg/ml gentamycin for 2 h. The cells were washed three times again with PBS and were further incubated with fresh medium. The infected cells and supernatant in each well were harvested at the indicated time intervals by washing the wells three times with sterilized distilled water. These bacterial suspensions were diluted in sterilized water and plated in known volume onto BCYE-α agar. The numbers of CFU in infected cells were counted at the indicated time points after infection.

Methods Enzymol 1987, 138:162–168 PubMedCrossRef 40 Payment P, T

Methods Enzymol 1987, 138:162–168.PubMedCrossRef 40. Payment P, Trudel M: Methods and Techniques in Virology.

New York: Marcel Dekker; 1993. Competing interests The authors declare that they have no competing interests. Authors’ contributions DW contributed to the study design, data collection, most experiments, writing of the initial draft, and revising the manuscript. WB, YW, WG, and RL collected the preliminary data, and helped to perform some experiments. ZY and NZ participated in the study design, interpretation of the data, the study coordination, technical issues, and revision of Selleckchem BV-6 the manuscript. All authors read and approved the final manuscript.”
“Background Due to the resistance against a wide range of antimicrobials including important ones such as penicillins and all cephalosporins [1], Extended Spectrum Beta-Lactamase (ESBL) find more producing bacteria are considered a vast threat to public health. Carriership of bacteria

producing ESBLs in humans is increasing in the community and health care. In Enterobacteriaceae ESBL-genes are mostly plasmid mediated and may be located on various plasmid types. In Dutch poultry bla CTX-M-1 is the predominant ESBL-gene, located on IncI1 plasmids [2] and these ESBL-genes seem to play an important SGC-CBP30 nmr role in humans as well [3]. The prevalence of ESBLs in poultry in the Netherlands is very high, 100% of investigated farms were positive for ESBL-producing Escherichia coli and on 85% of these farms, 80% (95% CI: 71-99%) or more of the animals carried ESBL-producers mafosfamide in their faeces [4]. Surveillance data show that among all broiler E. coli in the Netherlands, 15% carry plasmids with ESBL-genes [2]. The occurrence of the IncI1/CTX-M-1 combination in broilers as well as in humans indicates that the bacterium populations in poultry may play a role as a reservoir for ESBL-genes found in human

bacteria [5]. Although in general a high selective pressure by use of antimicrobials exists in broiler chickens, the reservoir role is unexpected in this particular case. Mass treatment of broiler chickens with cephalosporins is forbidden in the Netherlands. Cephalosporins are, however, used in one-day old reproduction animals in the poultry sector [6], selecting for bacteria producing ESBLs that can then successfully colonize broilers. To explain the widespread occurrence of the IncI1 and CTX-M-1 positive isolates, we wish to understand under what circumstances this gene-plasmid combination can be successful. The IncI1 plasmid is conjugative, and conjugation could explain the high abundance of bacteria carrying this plasmid in the microbiota of broilers. Within the microbiota, plasmids might act as infectious agents, which are able to persist by transfer to new bacterial hosts.

Hepatic veno-occlusive disease (VOD) is another recurrent complic

Hepatic veno-occlusive disease (VOD) is another recurrent complication after SC transplantation. VOD is a condition in which some of the small CB-5083 supplier hepatic veins are blocked, in this case, by cells. It is a complication of high-dose chemotherapy given before a BM transplant and it is marked by weight gain, due to fluid retention, increased liver size, and raised levels of bilirubin in the blood [101, 102]. VOD is more frequent in children undergoing SC transplantation [103].Two hundred and forty four HSCTs have

been evaluated and it has been found that VOD had appeared in 11% of them. It has been identified that risk factors for VOD are age <6.7 years, type of VOD prophylaxis, and busulphan-containing conditioning regimens [104]. Interesting results have been obtained in VOD treatment by oral defibrotide [105] and combination of intravenous heparin, oral glutamine and ursodiol [106]. Obstacles and possible

solutions The compatibility between the recipient and the graft is the main problem that must be faced off when a medical group decides to transplant organs, tissues or cells successfully. In SCT, the immunorejection also represents an important obstacle. If autogenous cells are available, immunorejection can be bypassed. In fact, common clinical practice is to harvest autogenous MCSs, expand them in culture, avoiding microorganism contamination, and store the obtained cell population before implantation [9]. Crenigacestat Interestingly, Mocetinostat molecular weight allogenic MCSs transplant, obviously applied in emergency situations, such as spinal cord injury or myocardial infarction, demonstrates high success rates. A tolerance of allogenic G protein-coupled receptor kinase MCSs seems to be induced by the same grafted cells. Indeed, MCSs inhibit T cell proliferation and maturation through direct cell-cell

effects and by secretion of soluble factors [107, 108]. Allogenous EC transplantation is not immunotolerated as MSCs graft. Therefore, avoiding the EC immunorejection, several strategies are being developed. Somatic cell nuclear transfer (SCNT) is currently the most promising of them. SCNT consists in the enucleation of the donor’s oocytes and the renucleation of them with nuclei taken from the patient’s somatic cells. The created cells are tolerated because they express major histocompatability complex (MHC) of the recipient. The disadvantages of SCNT include the creation and destruction of embryos and the current inability to apply the technology in autoimmune diseases [109]. In order to avoid autoimmune rejection, some elaborate methods, such as gene therapy, are under investigation [3, 110]. ESCs are characterized by genetic instability and imprinting genes dysregulation [111].

5 Gujarati F, 34 years (Gujarat region India; n = 71) 51% < 12 5

5 Gujarati F, 34 years (Gujarat region India; n = 71) 51% < 12.5 Solanki et al. [31] United Kingdom, Birmingham, end of winter. White buy BAY 80-6946 M, <65 years, mean 30 years men and women (n = 4) 28 ± 12 – White F, <65 years, mean 30 years men and women (n = 12) 48 ± 29 White M, >65 years, mean 74 years men and women (n = 4) 55 ± 14 White F, >65 years, mean 74 years men

and women (n = 14) 40 ± 21 Asian M, <65 years, mean 31 years men and women (n = 14) 16 ± 08 Asian F, <65 years, mean 31 years men and women (n = 3) 21 ± 07 Asian M, >65 years, mean 72 years men and women (n = 21) 13 ± 09 Asian F, >65 years, mean 72 years men and women (n = 16) 23 ± 20 Finch et al. [32] United Kingdom, London, all year round. White M (50%)+F, mean 39 years, Protein Tyrosine Kinase inhibitor winter (n = 30) 39 ± 18 Winter season (March/April), vegetarian,

Hindu religion, Muslim religion (only in winter); Hindus seasonal responses are blunted, resulting in significantly lower peak values than for whites PD98059 or non-vegetarian (Muslim) Asians White M (50%)+F, mean 39 years, summer (n = 18) 65 ± 27 Asian M (70%)+F, mean 42 years, non-vegetarians, winter (n = 116) 19 ± 13 Asian M (70%)+F, mean 42 years, non-vegetarians, summer (n = 22) 45 ± 24 Asian M (40%)+F, mean 42 years, vegetarians, winter (n = 29) 10 ± 8 Asian M (40%)+F, mean 42 years, vegetarians, summer (n = 16) 27 ± 21 Van der Meer et al. [1] The Netherlands, Amsterdam, The Hague, Amersfoort and Haarlem (52°N) Dutch M (40%)+F, median 45 years (n = 102) Median 67, 06% < 25 Autumn or winter season, pregnant or breastfeeding, lower consumption of fatty fish, no use of vitamin D supplements, smaller area of uncovered skin, no use of tanning bed, lower consumption of margarine, no preference for sun Surinam South Asian M (37%)+F, median 41 years (n = 107) Median 24, 51% < 25 Pregnant women Datta et al. [63] IMP dehydrogenase United Kingdom, Cardiff (51.5°N), at booking visit Indian subcontinent (n = 100) 52% < 20 Being in Britain for more than 3 years (compared to less than 3 years and to being born in Britain) Children Lawson and Thomas [40]

UK, autumn Bangladeshi M+F, 2 years (n = 139) 42 ± 21, 20% < 25 Failure to take a vitamin supplement. Pakistani M+F, 2 years (n = 200) 36 ± 20, 34% < 25 Indian M+F, 2 years (n = 279) 42 ± 23, 25% < 25 Koch and Burmeister [64] Germany, in summer Asian M (33%)+F, 3–17 years (Birma, Sri Lanka, India; n = 9) 28 ± 09, 44% < 25 – SD standard deviation a Unless mentioned otherwise Table 6 Studies among Indian populations in India Study Study characteristics Study population Serum 25(OH)D (nmol/l) Mean ± SD a Determinants for lower serum 25(OH)D Adults Goswami et al. [19] India, Delhi, in winter Adult M, mean 31 years (n = 244) 18 ± 9 – Adult F, mean 35 years (n = 398) 17 ± 11 Goswami et al. [41] India, Agota village (29° N), in winter Adult M, rural, mean 43 years (n = 32) 44 ± 24 Female gender Adult F, rural, mean 43 years (n = 25) 27 ± 16 Harinarayan et al.

Cardwell CR, Abnet CC, Cantwell MM, Murray LJ (2010)

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Compared with other screening techniques such as transcriptomics

Compared with other screening techniques such as transcriptomics or proteomics, SEREX offers a crucial advantage that subtle changes in the protein expressions can be detected through immunological reactions [32, 33]. Several authors have already applied SEREX to glioma, and some antigens, including glioma-expressed antigen 2 (GLEA2) [7], PHD finger protein 3 (PHF3) [7, 34], and SRY-box 6 (SOX6) [8] have been identified. It should be noted that we found autologous antibodies against SH3GL1 to be a low-grade glioma-specific marker with similar experimental systems to others. Our

unique approach was the quantitative comparison of the levels of serum antibodies using the ELISA, while the approach of others Inflammation related inhibitor was qualitative analysis. The application of ELISA in the validation step could lead to the discovery of a low-grade glioma-specific high titer of the I-BET-762 order autoantibody and the decrease in high-grade gliomas. Although some candidates of glioma biomarkers have been identified by various screening methods [6–8, 34–37], no serum marker for early diagnosis has been found yet. Therefore,

it is quite valuable to find a novel serum biomarker for its early diagnosis, prediction of the prognosis in each patient, and development of a new molecular learn more target. Indeed, The results of an overlap peptide array and ELISA using deletion mutants of SH3GL1 showed that 12 amino acids in the C-terminal portion, FPLSYVEVLVPL, were indicated as a major epitope site. By using a synthetic peptide corresponding to the epitope as an antigen, a more accurate screening for the patients

with low-grade gliomas and a specific peptide vaccine therapy would be achieved in the future. Author details 1Departments of Neurological Surgery, Chiba University, Graduate School of Medicine, 1-8-1, Inohana, Chuo-ku, Chiba 260-8670, Japan. 2Genetics and Biochemistry, Chiba University, Graduate School of Medicine, 1-8-1, Inohana, Chuo-ku, Chiba 260-8670, Japan. 3Diagnostic Pathology, Chiba University, Graduate School of Medicine, 1-8-1, Wilson disease protein Inohana, Chuo-ku, Chiba 260-8670, Japan. 4Department of Biochemistry, Graduate School of Life Science, Nagoya Women’s University, 3-40, Shioji-cho, Mizuho-ku, Nagoya 467-8610, Japan. References 1. Ohgaki H, Kleihues P: Epidemiology and etiology of gliomas. Acta Neuropathol 2005, 109:93–108.PubMedCrossRef 2. Anderson E, Grant R, Lewis SC, Whittle IR: Randomized Phase III controlled trials of therapy in malignant glioma: where are we after 40 years? Br J Neurosurg 2008, 22:339–349.PubMedCrossRef 3. van den Bent MJ, Afra D, de Witte O, Ben Hassel M, Schraub S, Hoang-Xuan K, Malmstrom PO, Collette L, Pierart M, Mirimanoff R, Karim AB: Long-term efficacy of early versus delayed radiotherapy for low-grade astrocytoma and oligodendroglioma in adults: the EORTC 22845 randomised trial. Lancet 2005, 366:985–990.PubMedCrossRef 4. Sanai N, Berger MS: Glioma extent of resection and its impact on patient outcome.

With the advancement of DNA-based biosensors and automation for b

With the advancement of DNA-based biosensors and automation for bacterial detection, enrichment broths could be screened for the presence of Campylobacter spp. in a shorter time, with greater sensitivity and without the generation of any microaerobic condition. In addition, food microbiology laboratories interested in establishing techniques

for the isolation of Campylobacter from retail meat will have access to a cost-effective enrichment procedure without the need to invest in systems to generate microaerobiosis. Reference documents from the FDA and FSIS USDA should eventually be updated to provide for an alternative, simplified protocol that yields similar number of Forskolin Campylobacter positive samples as the current

reference protocols. Methods Sample preparation, incubation and Campylobacter isolation Retail broiler meat samples (total = 108 samples; 49 breasts and 59 thighs) were purchased from local stores (Auburn, AL) from April 2009 to October 2010. Samples were tested in batches of three to five samples per week. Each meat package was considered one sample, and from each package ~1-inch pieces were cut aseptically and mixed thoroughly. For all samples, 25 g of meat was weighed two times (two subsamples) in individual, sterile Whirl-Pak® (Nasco, Fort Atkinson, WI). Each subsample was enriched in 100 ml of Bolton’s broth (with antimicrobial supplements) and 5% (v/v) of lysed horse blood [17]. The control subsamples (microaerobic

Enzalutamide price subsamples) were incubated in anaerobic jars gassed with a microaerobic gas mix (85% N2, 10% CO2, 5% O2; Airgas, Radnor, PA) using the evacuation-replacement system MACSmics Jar Gassing System (Microbiology International, Frederick, MD). The other subsamples (aerobic subsamples) were incubated without the addition of microaerobic gas mix, by closing the bags after removing the remaining air manually. All subsamples were incubated at 42°C for 48 h. After incubation and for all subsamples, 0.1 ml of the enriched broth was transferred Progesterone to modified charcoal cefoperazone deoxycholate agar [10] through a 0.65 μm membrane filter as described elsewhere [33]. All agar plates were incubated under microaerobic conditions at 42°C for 48 h. Presumptive Campylobacter colonies were observed under phase contrast microscopy (Olympus BX51, Olympus America Inc., Center Valley, P) for spiral morphology and darting motility. Presumptive isolates were Pictilisib price stored at -80°C in tryptic soy broth (Difco, Detroit, MI) supplemented with 20% glycerol (v/v) and 5% (v/v) lysed horse blood for further analysis. Identification of presumptive Campylobacter isolates by mPCR assays Campylobacter isolates were recovered from frozen stocks by transferring to Brucella agar plates supplemented with 5% horse blood and through 0.6 μm membrane filters as described above. Plates were incubated at 42°C under microaerobic conditions for 24 h.