The donor column refers to Typhimurium strains used as DNA sources for the transformation of E. coli TOP10 or DH5α. The SN-38 Xba I column indicates the check details Cluster name in which the donor strain was placed in the previously published PFGE- Xba I restriction dendrogram [16]. The CMY column denotes bla CMY-2-positive (+) and bla CMY-2-negative (-) plasmids. The phenotype column describes the resistance phenotype of the donor strain and

the resistances transferred by the IncA/C plasmids (underlined). The abbreviations for the antibiotics are described in Methods. The estimated plasmid sizes are indicated in terms of bp. The next ten columns display the results of the PCR screening scheme (Additional file 1, Table S1, Figure 3, Figure 4 and Methods). Positive amplifications are designated by a plus symbol (+) and negative amplifications by a minus symbol (-). In the case of the IP-1 and floR columns, the + (-) code indicates that the Typhimurium donor strain was positive for the marker but that the E. coli transformants were negative. “”1 kb”" indicates an integron of around 1,000 bp amplified in pAR060302, as previously described by Call et al. selleck [6]. nd, not determined. Characterization of IncA/C plasmids based on the antibiotic resistance phenotype To isolate and characterize the IncA/C plasmids present in the Mexican ST213 genotype, E. coli TOP10 or DH5α transformants were obtained

using plasmid DNA isolated from 32 CMY+ and 13 CMY- strains. Ceftriaxone was used to select CMY+ plasmids, and chloramphenicol was used to select CMY- plasmids because this resistance has been found to be part of the IncA/C plasmid backbone [5, 6, 8]. All the transformants carrying the IncA/C plasmids also displayed resistance to ampicillin,

chloramphenicol, sulphonamides, streptomycin and tetracycline. Resistance to gentamicin was conferred by most of the CMY+ plasmids, and trimethoprim-sulfamethoxazol resistance was mostly detected in the plasmids containing PtdIns(3,4)P2 the IP-1 integron (dfrA12, orfF and aadA2; see below). Resistance to neither kanamycin nor nalidixic acid was transferred (Figure 2). These results indicate that the MDR phenotypes of ST213 strains can be explained largely by the presence of IncA/C plasmids. Pst I restriction fingerprints The plasmid profiles showed that all of the E. coli transformants carried one large plasmid of between 100 and 160 kb. These transformants were analyzed by Pst I restriction fingerprinting [12, 23]. Cluster analysis of the Pst I fingerprints showed two main plasmid types (similarity <50%), which we named type I and type II (Figure 2). All of the CMY+ plasmids were contained in type I and were distributed into three clusters (a, c and d). The CMY- plasmids were found in two distinct groups: one in type II and the other in cluster b within type I, suggesting that the CMY- plasmids originated from two divergent IncA/C plasmid types.

The absence of an increase in SCr VX809 levels after the administration of NAC does not always indicate that NAC is effective in preventing CIN. NAC is known to increase the activity of creatinine kinase and the excretion of creatinine from the renal tubules [141, 142]. Accordingly, it cannot be concluded that NAC may preserve kidney function even when no increase in SCr levels is observed after treatment with NAC,

because NAC may maintain the patient’s baseline SCr level by increasing excretion of SCr. Although the use of NAC is not Blasticidin S research buy recommended for a measure to prevent CIN, some specialists recommend it for high risk patients because of the low cost and low incidence of adverse drug reactions [8, 143]. Does hANP decrease the risk for developing CIN? Answer: We consider not to use hANP to prevent CIN. An intrinsic peptide, hANP exerts a natriuretic action, afferent arteriole dilatation [144], anti-renin and anti-aldosterone actions [145], and has been reported to be beneficial in the treatment of AKI after cardiac surgery [146]. Although several reports have denied the efficacy of hANP in preventing CIN [147–149], the decrease in blood pressure by hANP might have affected the incidence

of CIN in these reports. A study in Japan has reported that hANP at a low dose that does www.selleckchem.com/products/fosbretabulin-disodium-combretastatin-a-4-phosphate-disodium-ca4p-disodium.html not decrease blood pressure is beneficial in the prevention of CIN [150]. However, there is no conclusive evidence supporting the efficacy of hANP in preventing CIN, and at the present time, hANP is not recommended as a standard measure to prevent CIN. Further studies are awaited to investigate the indications of hANP in the prevention of CIN in high risk patients. Sclareol B-type natriuretic

peptide (BNP) is also expected to be effective in the prevention of CIN, and further studies are awaited to evaluate its efficacy [151]. Does ascorbic acid decrease the risk for developing CIN? Answer: We consider not to use ascorbic acid to prevent CIN. Ascorbic acid exerts an anti-oxidant action against reactive oxygen species, and potentiates the effects of other antioxidants [152, 153]. Spargias et al. [152] have reported the efficacy of ascorbic acid in preventing CIN. In the REMEDIAL study in which 326 patients with CKD were randomly assigned to prophylactic administration of 0.9 % saline infusion plus NAC, sodium bicarbonate infusion plus NAC, or 0.9 % saline plus ascorbic acid plus NAC, ascorbic acid was not effective in the prevention of CIN [154]. At the present time, the use of ascorbic acid is not recommended as a standard measure to prevent CIN. Do statins decrease the risk for developing CIN? Answer: We consider not to use statins to prevent CIN. Because statins exert many different actions, including anti-oxidant and anti-inflammatory actions [155], they are expected to be effective in preventing CIN.

These cases were divided into two groups: group 1 (HCC; n = 35), samples were collected from patients diagnosed and treated at the National Cancer Institute, Cairo University, between December 2005 and August 2008; group 2 (CH; n = 34), samples were collected from HCV associated chronic hepatitis (CH) patients admitted to Kasr Al-Aini School of Medicine, Cairo University, in the same period

and enrolled in routine diagnosis or therapeutic procedures. The mean age of CH patients was 47.5 years and M:F ratio was 1.5:1, whereas the mean age of HCC was 51.6 years and M:F ratio was 1.3:1. All cases of CH were graded and staged according to the modified Knodell scoring system [23] and all HCC selleck chemicals cases were graded according to the World Health Organization (WHO) classification criteria and staged according to the American Joint Pritelivir committee on Cancer [24]. The percent of normal to tumor ratio were more than 80% in all studied cases to overcome the nominalization effect of the tumor stroma and/or necrosis as well as the cirrhotic tissues factors in the studied specimens. Table 1 illustrates the clinico-pathological features of the studied cases. Normal liver tissue samples this website were obtained from liver transplant donors (15 samples) and were used as controls. A written consent was obtained from

all patients and normal liver donors prior to enrollment in the study and the ethical committee of NCI approved the protocol, which was in accordance with the ethical guidelines of the 1975 Declaration of Helsinki. Table 1 Clinical features of the studied groups of patients. Variables HCC CH   n = 35 (%) n = 34 (%) Liver Function Test (Mean ± SD)     ALT 77.2 ± 76.2 74.33 ± 30.97 AST 70.577 ± 49.4 81.66 ± 35.35 Alk ph 181.1 ± 174.2 111.57 ± 61.58 Alb 3.758 ± 0.707 3.9 ± 0.538 T.Bil 1.1846 ± 0.523 1.34 ± 0.897 INR Methamphetamine 1.179

± 0.067 1.22 ± 0.161 Complete Blood Picture (Mean ± SD)     Hb 12.3 ± 1.64 13.59 ± 2.24 TLC 6.186 ± 3.163 6.509 ± 2.05 Plt 177 ± 121 175.5 ± 67.267 Viral marker     HBs-Ag 0 (0) 0 (0) HCV-Ab 35 (100) 34 (100) HBV-PCR 0 (0) 0 (0) HCV-PCR 35 (100) 34 (100) Tumor Marker (Mean ± SD)     Serum AFP 1885 ± 5888 265 ± 110 AFP, alpha fetoprotein; Alb, albumin; Alk, Alkaline Phosphates; ALT, alanine aminotransferase; CH, chronic hepatitis; Hb, hemoglobin; HBs-Ag, hepatitis B surface antigen; HCC, hepatocellular carcinoma; HCV, hepatitis C virus; INR, International normalized ratio; PCR, polymerase chain reaction; Plt, platelet count; TLC, total leukocytic count; T.Bil, total bilirubin. HepG2 cell culture HepG2 cells were used to establish the in vitro HCV replication. HepG2 culturing and infection were carried out according to previous protocols [25]. Briefly, HepG2 cells were maintained in 75 cm culture flasks (Greiner bio-one GmbH, Germany) containing Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 4.

2 Adequacy of the genetic risk perception Overestimation 77 66.9 Adequate Estimation 30 26.1 Underestimation 8 6.9 *14 subjects were unable to report their risk levels for cancer of the breast and/or ovaries **15 subjects were unable to report their level of risk of being a carrier of the genetic mutation of the BRCA1 and BRCA2 genes Subjective and objective risk The mean percentage regarding the subjective risk of developing a tumour and of being a carrier of the genetic mutation were 39% and

40%, respectively. The mean percentage regarding the objective risk, selleck chemicals calculated using the BRCAPRO model, of developing a tumour and of being a carrier of the genetic mutation were 11% and 19%, respectively. Anxiety and Depression The total mean score was 13, with 24% of the find more subjects suffering one episode of

major depression and 19% experiencing the presence of some disturbance in adaptation. A mean score of 8 was found for the single scales (borderline anxiety) and of 5 (normal depression). A total of 25% had borderline anxiety levels and the same value was found in subjects suffering from anxiety. Depression was found in 9% of the subjects, while 15% were borderline. Association between medico-demographic variables and PND-1186 risk perception (table 4 and 5) Table 4 Associations between the perception of risk (CRP-GRP) and Medical-Demographic variables   N Mean Std. Deviation P (2-tailed) ELIGIBILITY Cancer Risk Perception         Non-Eligible 44 32.82 21.87   Eligible 72 43.04 24.13 .024* Genetic Risk Perception         Non-Eligible 43 29.11 21.92   Eligible 72 46.45 21.96 .000* PATHOLOGY Carnitine palmitoyltransferase II Cancer Risk Perception         Non-Affected 84 38.63 21.14   Affected 32 40.89 30.35 .712 Genetic Risk Perception         Non-Affected 83 37.90 22.99   Affected 32 45.23 23.74 .108 Table 5 Associations

between the perception of risk (CRP-GRP) and Medical-Demographic and Psychological variables   Cancer risk perception Genetic risk perception Anxiety        Pearson coefficient 0.050 0.087    P (2-tailed) 0.596 0.355 Depression        Pearson coefficient -.031 .072    P (2-tailed) .742 .537 Age        Pearson coefficient -.068 -.030    P (2-tailed) .468 .747 Number of relatives affected by breast and/or ovarian cancer        Pearson coefficient .053 -.082    P (2-tailed) .569 .386 Number of relatives affected by other types of tumour        Pearson coefficient -.149 -.139    P (2-tailed) .111 .140 BRCA pro Cancer Risk        Pearson coefficient .254      P (2-tailed) .006 — BRCA pro Genetic Risk        Pearson coefficient   .322    P (2-tailed) — .000 Of all the medical-demographical variables, only the condition of eligibility was found to be statistically associated to the perception of risk (Table 4). The subjects who were eligible for genetic testing had a significantly higher perception of risk compared to the non-eligible people (CRP = 43%vs33%, p = 0.024; GRP = 46%vs29%, p < 0.000).

They are thus exported from the cell via the transporter-mediated system [140]. Basing on these data we can affirm

that GST can lead to a loss of response to chemotherapeutic agents, including those that are usually employed in the treatment of ovarian cancer. Some authors let us think that this is particularly significant in cancer cells with stem-cell like properties. In a recent study, it has been shown that platinum-resistant human cancer cells with stem-cell like EMT properties, had high cellular GSH and accumulated significantly less cellular platinum compared to their parental cells, and failed to undergo apoptosis when exposed to platinum at the drug concentrations toxic to the parental cells [140]. Apoptosis Apoptosis can condition response to antitumor drugs and it’s regulated by several molecular phenomena, such as the expression of Bm-1 and the loss of p53. Bmi-1, selleck a member of the polycomb group (PcG) family, participates in the self-renewal and maintenance of CSCs [141]. As an oncogene, Bmi-1 could enable cancer cells to escape apoptosis by modulating multiple growth signaling pathways [142]. Thus, its overexpression in cancer cells could be used as a survival marker. The

role of Bmi-1 in chemoresistance has been addressed recently [143, 144]. For ovarian cancer cells, silencing of Bmi- 1 gene could promote sensitivity to cisplatin and induction of apoptosis [145]. The tumor suppressor see more gene p53 plays a critical role in cell learn more proliferation and apoptosis by controlling several signaling pathways. In addition, the control of intracellular localization of p53 is also associated with the regulation of apoptosis and chemosensitivity in human ovarian Quisqualic acid cancer cells

[146–148]. Loss of p53 function correlates with multidrug resistance in several tumor types, including EOC [149]. Enrichment of CSCs during disease progression Enrichment of CSCs in tumor tissues is reported in patients with response to therapy through mechanisms such as enhanced DNA damage repair and changes in the cellular phenotype between epithelial and mesenchymal states of cell [150]. EMT is a physiological transcriptional reprogramming event and is characterized by the combined loss of epithelial cell junctions and cell polarity and the gain of a mesenchymal phenotype. EMT and mesenchymal to epithelial transition (MET) processes are now recognized in cancer progression [151]. A link between CSC and EMT has been suggested, whereby transformed human mammary epithelial cells, that have undergone EMT, show a gain of the CSC phenotype [152–155]. Recently, Kurrey et al. have reported a detailed study of genome-wide identification of SNAI1 and SNAI2 targets that resolves the specific mechanism underlying enrichment of stem-like cells post radiation treatment or chemotherapy through EMT [156].

The pattern

of Chromatocurvus halotolerans DSM 23344T was characterized by an aminophospholipid and an unidentified phospolipid in addition to the dominating polar lipids phosphatidylglycerol and phosphatidylethanolamine (Table  1), so that it could be distinguished from the profiles of Ivo14T, H. rubra and C. litoralis. However, the profile of Chromatocurvus halotolerans did match the polar lipid patterns of type strains of the chemoheterotrophic species H. salexigens and H. mediterranea that were obtained in this study and differed slightly from results published elsewhere [17, 19]. The whole-cell fatty acid patterns of the strains Ivo14T, Chromatocurvus halotolerans CP673451 datasheet DSM 23344T and H. rubra DSM 19751T were determined upon growth

on Marine Agar 2216 plates. The results were compared with the cellular fatty acid profiles of the type strains of C. litoralis and two related chemoheterotrophic Haliea species (Table  2). The fatty acid pattern of H. rubra DSM 19751T could be distinguished from all other type strains by the low content of 17:0, 17:1 and 10:0 3OH fatty acids, whereas C. litoralis DSM 17192T was unique in the synthesis of the unusual 16:1 ω6 unsaturated fatty acid, which suggests an affiliation of both type strains to different genera. Further analyses of the cellular Peptide 17 ic50 fatty acid profiles of the four BChl a-containing strains were performed upon cultivation in SYPHC liquid medium with different oxygen concentrations in the head space gas atmosphere (see Additional file 1). In a previous study it was found that in C.

litoralis the position of the double bond in the unsaturated fatty acids 16:1 and 18:1 depends on the oxygen saturation and was shifted from the ω7 to the ω6 position under conditions of oxygen limitation [8]. It is known that several pathways for the synthesis of unsaturated fatty acids exist in proteobacteria. selleck inhibitor An oxygen-dependent pathway is based on desaturases that introduce double bonds in membrane-bound fatty acids by oxidation with molecular oxygen. An alternative oxygen-independent pathway introduces double bonds during elongation of the fatty acid chain [35]. Hence, we propose that C. litoralis expresses two distinct desaturases for the fatty acids 16:1 ω7 (Δ9 desaturase, JNJ-64619178 encoded by the proposed gene KT71_07544) and 18:1 ω7 (Δ11 desaturase, probably encoded by KT71_03222), whereas the ω6 unsaturated fatty acids are produced by an oxygen-independent pathway. A similar effect could not be detected in the strains Ivo14T, Chromatocurvus halotolerans DSM 23344T and H. rubra DSM 19751T (Additional file 1). While in the analyzed fatty acid patterns of strain Ivo14T neither the abundance of the unsaturated fatty acids 18:1 ω7 nor 16:1 ω7 correlated with the oxygen saturation, in Chromatocurvus halotolerans a decrease of the portion of 18:1 ω7 from 36.6% to 25.8% under conditions of oxygen limitation was detected, which indicates involvement of an oxygen-dependent desaturase.

Br J Haematol 2004,125(6):749–755.PubMed 160. Eisenbarth GS: Update in type 1 diabetes. J Clin Endocrinol Metab 2007,92(7):2403–2407.PubMed 161. Aiello LP, Gardner TW, King GL, Blankenship G, Cavallerano JD, Ferris FL, Klein R: Diabetic retinopathy. Diabetes Care 1998,21(1):143–156.PubMed 162. Sima AA, Zhang W, Grunberger G: Type 1 diabetic neuropathy and C-peptide. Exp Diabesity Res 2004,5(1):65–77.PubMed 163. Ingberg

CM, Palmer M, Schvarcz E, Aman J: Prevalence of urinary tract symptoms in long-standing type 1 diabetes mellitus. Diabetes Metab 1998,24(4):351–354.PubMed 164. Couri CE, Oliveira MC, Stracieri AB, Moraes DA, Pieroni F, Barros GM, Madeira MI, Malmegrim KC, Foss-Freitas MC, Simoes BP, et al.: C-peptide levels and insulin independence following autologous nonmyeloablative hematopoietic stem cell transplantation in newly diagnosed type 1 diabetes mellitus. JAMA 2009,301(15):1573–1579.PubMed 165. Snarski E, Torosian T, Paluszewska BAY 11-7082 molecular weight M, Urbanowska E, Milczarczyk A, Jedynasty K, Franek E, Jedrzejczak WW: Alleviation of exogenous insulin requirement in type

1 diabetes mellitus after immunoablation OTX015 mw and transplantation of autologous hematopoietic stem cells. Pol Arch Med Wewn 2009,119(6):422–426.PubMed 166. Trivedi HL, Vanikar AV, Thakker U, Firoze A, Dave SD, Patel CN, Patel JV, Bhargava AB, Shankar V: Human adipose tissue-derived mesenchymal stem cells combined with hematopoietic stem cell transplantation synthesize insulin. Transplant Proc 2008,40(4):1135–1139.PubMed 167. Wijesekera LC, Leigh PN: Amyotrophic lateral sclerosis. Orphanet Farnesyltransferase J Rare Dis 2009, 4:3.PubMed 168. Janson CG, Ramesh TM, During MJ, Leone P, Heywood J: Human intrathecal transplantation of peripheral blood stem cells in amyotrophic lateral sclerosis. J Hematother Stem Cell Res 2001,10(6):913–915.PubMed 169. Mazzini L, Ferrero I, Luparello V, Rustichelli D, Gunetti M, Mareschi K, Testa L, Stecco A, Tarletti R, Miglioretti M, et al.: Mesenchymal stem cell transplantation

in amyotrophic lateral sclerosis: A Phase I clinical trial. Exp Neurol 2010,223(1):229–37.PubMed 170. Mazzini L, Fagioli F, Boccaletti R, Mareschi K, Oliveri G, Olivieri C, Pastore I, Marasso R, Madon E: Stem cell therapy in amyotrophic lateral sclerosis: a methodological approach in humans. Amyotroph Lateral Scler Other Motor Neuron Disord 2003,4(3):158–161.PubMed 171. Martinez HR, Gonzalez-Garza MT, Moreno-Cuevas JE, Caro E, Gutierrez-Jimenez E, Segura JJ: Stem-cell transplantation into the frontal motor cortex in amyotrophic lateral sclerosis patients. Cytotherapy 2009,11(1):26–34.PubMed 172. Papadeas ST, Maragakis NJ: Advances in stem cell research for Amyotrophic Lateral Sclerosis. Curr Opin Biotechnol 2009,20(5):545–551.PubMed 173. Astradsson A, Cooper O, Vinuela A, Isacson O: Recent advances in cell-based therapy for Parkinson disease. selleck Neurosurg Focus 2008,24(3–4):E6.PubMed 174. Weintraub D, Comella CL, Horn S: Parkinson’s disease–Part 2: Treatment of motor symptoms.

In the Zn1−x Cu x O nanostructures, the presence of the E2(high) mode confirms that they all have a typical hexagonal wurtzite structure, which is consistent with the above HRTEM and XRD observations. When the Cu content is 7%, the E2(high) and E1(LO) modes become broader and shift to lower frequency, as compared with the undoped counterpart. This may be due to the decrease in the binding energies of Zn-O bonds as a result of the Cu

doping, indicating that the long-range order of the ZnO crystal is destroyed S63845 datasheet by Cu dopants [32]. Figure 5 Raman spectra. Raman spectra of undoped ZnO and Zn1−x Cu x O samples with the Cu contents of 7%, 18%, and 33%. On the other hand, three additional modes at around 290, 340, and 628 cm−1 can be observed. They are attributed to the Ag, B1 g, and B2 g modes of CuO due to the vibrations of oxygen atoms, respectively [33, 34]. From Figure 5, it is obvious that the intensity of the CuO peaks enhanced while that of ZnO

peaks decreases with the Cu concentration increases up to 33%. Such behavior is caused by the competition of Zn and Cu during the oxidization process. In the sample with the highest Cu content of 33%, the formation of CuO is dominant, in spite of the fact that the lower melting point and higher vapor pressure of Zn than those of Cu under the same conditions [35]. The formation of CuO is significant to induce the usual ZnO hexagonal structures changing into four-folded find more cross-like structures, in good agreement with the growth VX-689 manufacturer mechanism we have proposed above. In order to investigate the effects of the different Cu concentrations on the optical characteristics in the yielded samples, we have carried out PL spectroscopy

as shown in Figure 6. We can see that all the samples show two emission peaks: a sharp one appearing at approximately 377 nm in the ultraviolet (UV) region and another broad one in the visible region. The former is ascribed to the near-band-edge (NBE) exciton recombination, while the latter is quite complicated due to the native and dopant-induced defects Dynein in ZnO. The intensive PL emission peak at 495 nm is suggested to be mainly due to the presence of various point defects, which can easily form recombination centers. The peak corresponding to 510 nm is usually generated by the recombination of electrons in singly ionized oxygen vacancies with photogenerated holes in the valence band [36, 37]. Apart from the strong peaks at 495 and 510 nm, the visible band consists of at least four sub-peaks at wavelengths of 530, 552, 575, and 604 nm, resulting from the local levels in the bandgap of ZnO. The green shoulders at 530 and 552 nm are attributed to the antisite oxygen and interstitial oxygen, respectively [35]. The peak at 604 nm is possibly caused by the univalent vacancies of zinc in ZnO. The origin of another peak at 575 nm has been rarely mentioned and is still unclear.

The internal fragment was digested by HindIII and BamHI and subsequently cloned into pUCerm [24] to obtain a PD0325901 mouse plasmid designated pUCerm::covS. For electroporation, thawed electrocompetent cells (100 μl) were initially mixed on ice with 10 μl pUCerm::covS. The mixture was next transferred to a pre-chilled 2 mm electrode spacing cuvette (Bio-Rad). Electroporation was then performed using

a Gene Pulser II electroporator (Bio-Rad) with the following settings: voltage 1750 V, capacitance 25 μF, 12 ms, 481Ω. Subsequently, 1 ml pre-warmed Todd-Hewitt broth (Invitrogen) supplemented with 0.5% yeast extract and 0.125 M sucrose was added to the transformed cells, and the suspension was incubated at 37°C for 2 h. Transformants

PKA activator were selected on THB agar plates supplemented with 0.5% yeast extract and 5 μg/ml erythromycin. Successful integration of the plasmid was confirmed by PCR analysis www.selleckchem.com/products/idasanutlin-rg-7388.html of junction fragments using standard protocols (for the used primer locations please refer to fig. 1 and the result section). The generated insertional mutant strains were designated M18::covS, M18_588::covS, M49::covS, M49_581::covS, M49_634::covS, M2::covS, M2_583::covS, M6::covS, M6_586::covS, M6_796::covS and M6_576::covS. Figure 1 Schematic representation of the inactivation of CovS. A. Inactivation of CovS in the serotype M49 strain 591. Plasmid pUCerm::covS contains a fragment internal of covRS and confers erythromycin resistance (EmR). The genomic regions from both covR and covS used for recombination Cepharanthine are marked in black. B. M49 covRS locus after insertion of the plasmid pUCerm::covS. The thin arrows depict primers used for RT-PCR analysis (see below). The thick numbered arrows (1-4) represent primers used for PCR of whole region and junction fragments to confirm plasmid integration into the chromosome. C. RT-PCR analysis. Primer pairs derived from covR and covS were used. Lane DNA Ladder, O’GeneRuler 1 kb DNA Ladder (Fermentas); gDNA, genomic DNA; cDNA, first-strand synthesized cDNA; mRNA, messenger RNA; -C, negative control, where no template for polymerization

was used. From each GAS serotype under investigation a WT and mutant strain pair was tested for unaltered growth phenotypes in regular batch cultures using THY and BHI medium (additional file 1) Eukaryotic cell adherence For all adherence studies the HaCaT cell line was used, which is a spontaneous immortalized human keratinocyte cell line [25], obtained from German Cancer Research Center, Heidelberg, Germany. The adherence assay was performed as described previously [26]. In brief, all GAS strains were grown in THB supplemented with 0.5% yeast extract at 37°C under a 5% CO2 -20% O2 atmosphere. After overnight incubation the bacterial cells were suspended in modified Eagle’s medium supplemented with 10% fetal calf serum and added to 3.

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