However, in the near future, investigation of a larger cohort or a population-based analysis of the rate of each renal disease may reveal the Tipifarnib mouse actual frequency of the disease and the distribution of age ranges by utilizing the J-RBR system. Acknowledgments The authors greatly acknowledge the help and assistance of many

colleagues in centers and affiliated hospitals with collecting the data. We also sincerely thank Ms. Mayumi Irie in the UNIN-INDICE for establishing and supporting the registration system of J-RBR. This study was supported by the committee grant from the Japanese Society of Nephrology and by a grant-in-aid from the Research Group on Progressive Renal Disease

from the Ministry of Health, Labor and Welfare, Japan. 17-AAG datasheet electronic supplementary material Below is the link to the electronic supplementary material. Supplementary Table (DOC 38 kb) Appendix The following investigators participated in the project for developing the J-RBR: Hokkaido District KKR Sapporo Medical Center (Pathology), Akira Suzuki. Tohoku District Tohoku University Hospital and affiliated hospitals (Internal Medicine), Keisuke Nakayama, Takashi Nakamichi. Kanto District Chiba-East National Hospital (Clinical Research Center), Takashi DNA-PK inhibitor Kenmochi, Hideaki Kurayama, Motonobu Nishimura; The Jikei University Hospital (Internal Etoposide ic50 Medicine); Tokyo Metropolitan

Kiyose Children’s Hospital (Pediatric Nephrology), Hiroshi Hataya, Kenji Ishikura, Yuko Hamasaki; Tokyo Women’s Medical University Hospital (Pediatric Nephrology), Ishizuka Kiyonobu; Tsukuba University Hospital (Pathology and Nephrology), Joichi Usui. Koushinetsu District Niigata University Medical and Dental Hospital (Internal Medicine), Naofumi Imai; Shinshu University Hospital (Internal Medicine), Yuji Kamijo, Wataru Tsukada, Koji Hashimoto. Hokuriku District Kanazawa Medical University Hospital (Internal Medicine), Hiroshi Okuyama, Keiji Fujimoto, Junko Imura; Toyama Prefectural Central Hospital (Internal Medicine), Junya Yamahana, Masahiko Kawabata. Tokai District Nagoya University Hospital and affiliated hospitals (Internal Medicine), Japanese Red Cross Nagoya Daini Hospital (Kidney Center), Asami Takeda, Keiji Horike, Yasuhiro Otsuka. Kinki District Kyoto University Hospital (Internal Medicine); Osaka Kaisei Hospital (Pathology) and Osaka University Hospital (Internal Medicine), Yoshitaka Isaka, Yasuyuki Nagasawa, Ryohei Yamamoto; Wakayama Medical University Hospital (Pediatrics), Koichi Nakanishi, Yuko Shima. Chugoku District Kawasaki Medical School (Internal Medicine), Naoki Kashihara, Takehiko Tokura; Okayama University Hospital (Internal Medicine), Masaru Kinomura, Hiroshi Morinaga, Tatsuyuki Inoue.

Nine patients showed clinical PR, 10 showed 3-MA purchase SD, and 2 showed PD. The clinical response rate (CR or PR) of the neck disease was 42.9%. Table 4 Clinical response of the neck disease   CR PR SD PD Response rate Level 1   1 1   50% Level 2     1 1 0% Level 3   1 2   33.3% Level 4   2 1   66.7% Level 5     3   0% Level 6   1 1 1 33.3% Level 7  

3     100% Level 8   1 1   50% Total   9 10 2 42.9% Abbreviations: CR = complete response, PR = partial response, SD = stable disease, PD = progressive disease After surgery, local failure developed in one patient (level 6), and neck failure and distant metastasis occurred in another (level 7). With a median follow-up of 67 months, the 5-year overall survival rate was 90.0%, and the 5-year cumulative survival was 93.1%. Discussion We set out to determine the safety and

reliability of concurrent S-1 and radiotherapy in advanced selleckchem cancer of the oral cavity, in a phase I study. Many studies have demonstrated that combined chemotherapy AZD5582 supplier and radiation is a highly effective treatment modality for increasing the survival of patients with advanced disease [2, 3, 9–11]. Concurrent chemoradiotherapy has been established as an appropriate standard for many patients with locally advanced head and neck cancer. To the best of our knowledge, this study is the first trial of S-1 and radiotherapy in oral cancer. Tsukuda et al. reported that most adverse events of S-1 administration alone were hematological, Glycogen branching enzyme gastrointestinal, and skin toxicities, although most of these were grade 1 or 2 and controllable [12]. In the present study, there was no severe hematological, gastrointestinal, or skin toxicity. Mucositis was the most common adverse event, with grade 3 mucositis observed in 66.7% of patients at levels 5, 6, and 7 (Additional file 1). Grade 4 mucositis, constituting DLT, was observed in 2 of 6 patients at level 8. The doses used level 8 was deemed the MTD. Therefore, the determined recommended dose of S-1 was the reduced dose for 5 days

per week for 4 weeks (level 7). In a multi-institutional cooperative late phase II clinical study of S-1 alone in patients with advanced/recurrent head and neck cancer in Japan, the clinical response rate of the primary tumor was 36.4% in oral cancer patients [13]. In the present study, the overall clinical response rate was 93.3%, and the histological response rate was 90.0%, appearing to be remarkably good. Many studies have demonstrated concurrent chemoradiotherapy to be effective in patients with advanced head and neck cancer. However, the majority of studies have reported total radiation doses of more than 60-Gy. Tsukuda et al. reported that the complete response rate were 93% in stage III and 54% in stage IV, by treating head and neck cancer with S-1 and radiotherapy at a total dose of 66-70.2 Gy [14]. There have been few reports on the effect of preoperative chemoradiotherapy with a total radiation dose of 40-Gy [2, 3].

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44. Thompson JD, Higgins DG, Gibson TJ: Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight-matrix choice. Nucl Acids Res 1994, 22:4673–4680.PubMedCrossRef 45. Notredame C, Higgins DG, Heringa J: T-Coffee: A novel method for fast and accurate multiple sequence alignment. J Mol Biol 2000, 302:205–217.PubMedCrossRef 46. Garnier J, Gibrat JF, Robson B: GOR method for predicting protein secondary structure from amino acid sequence. Methods Enzymol 1996, 266:540–553.PubMedCrossRef 47. Cole C, Barber JD, Barton GJ: The Jpred 3 secondary structure prediction server. Nucleic Acids Res 2008, 36:W197–201.PubMedCrossRef 48.

PubMedCrossRef 44. Wani RA, Parray FQ, Bhat NA, Wani MA, Bhat TH, Farzana F: Non traumatic terminal ileal perforation. World J Emerg Surg 2006, 1:7.PubMedCrossRef 45. Urassa M, Isingo R, Kumogola Y, Mwidunda P, Helelwa M, Changulucha J, Mngara J, Zaba B, Calleja T, Slaymaker E:

Effect of PMTCT availability on choice of ANC in Mwanza and Magu districts and its impact on HIV sentinel surveillanc. Tanzania: Report of ANC surveillance Mwanza and Magu Districts 2007. 46. Beniwal US, Jindal D, Sharma J, Jain S, Shyman G: JPH203 price Comparative study of operative procedures in typhoid selleckchem perforation. Indian J Surg 2003, 65:172–7. 47. Kella N, Radhi PK, Shaikh AR, Leghari F: Qureshi MA: Factors affecting the surgical outcome in typhoid intestinal perforation in children. Paed Surg 2010,16(4):567–570. 48. Kaybal I, Gokcora IH, Kaybal M: A contemporary evaluation of enteric perforation in typhoid fever; analysis of 257 cases. Int Surg YH25448 cost 1990, 75:96–100. 49. Elesha SO: Pathology and pathogenesis of typhoid fever. Nig P Med J 1994, 1:38. 50. Shah AA, Wani KA, Wazir BS: The ideal treatment of typhoid enteric perforation- resection anastomosis. Int Surg 1999, 84:35–8.PubMed 51. Mawalla B, Mshana SE, Chalya PL, Imirzalioglu C, Mahalu W: Predictors of surgical site infections among patients undergoing major surgery at Bugando Medical Centre in Northwestern Tanzania. BMC Surgery

2011, 11:21.PubMedCrossRef 52. Karmacharya B, Sharma VK: Results of typhoid perforation management: our experience in Bir Hospital, Nepal. Kathmandu University Med J 2006, 4:22–24. 53. Meier DE, Tarpley JL: Typhoid intestinal

perforations in Nigerian children. World J Surg 1998, 22:319–323.PubMedCrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions PLC contributed in study design, literature search, data analysis, manuscript writing, editing and submission of the manuscript. JBM, MK, HJ, SEM, MM and GG participated in study Selleck Sunitinib design, data analysis, manuscript writing & editing. MDM participated in data analysis, literature search, manuscript writing & editing. JMG supervised the study and contributed in data analysis, manuscript writing & editing. All the authors read and approved the final manuscript.”
“After years of initial aggressive surgical intervention and a subsequent shift to damage control surgery (DCS), non operative management (NOM) has been shown to be safe and effective. In fact trauma surgeons realized that in liver trauma, it was safer to pack livers [1] than do finger fracture [2] or resection, and this represented a tangential issue to nonoperative approach. Damage control was not the paradigm shift for spleen and liver, but rather to address coagulopathy that was more commonly associated with penetrating major abdominal vascular injuries [3].

The number of expressed MTases in H. pylori strains was high, as reported [18, 26, 27, 29, 30], with a total average of 15.8 ± 2.2, (range 9-20), among 27 tested REases (isoschizomers excluded). Selection of methyltransferases with non-random geographic distribution A chi-square independence test was used to select the independent variables to be applied in the logistic regression models (Additional file 2: Table S3). Ten MTases were associated with the geographic origin of the strains analysed. A significant result was determined by the

analysis of standardized residuals (std. residual) for all MTases presenting a geographic association, except M. MspI and M. TaqI (Table 1). A Fischer test was applied and all significant MLN2238 mw associations were confirmed (Additional file 2: Table S4). Table 1 MTases presenting a statistical significant BI 2536 ic50 association selleck chemicals llc with isolates of distinct geographic origin (Chi-square test). MTase Recognition sequence * Chi-square higher smaller Std.     (p value) expression in isolates from Residual M. AseI ATTAAT 0.031 — Africa 2.13 M. FokI GGATG 0.001 America Asia — 2.77 2.55 M. MspI CCGG 0.036 — –   M. Hpy188I TCNGA 0.002 America — 2.05 M. Hpy99I CGWCG 0.025 America — 2.29 M. HpyCH4III CANGT <0.001 Africa America -- -1.99 -2.21 M. DraI TTTAAA

<0.001 Asia -- 5.36 M. BstUI CGCG 0.006 Asia -- 2.81 M. FauI CCCGC 0.004 Asia -- -2.04 M. TaqI TCGA 0.044 -- --   * data from REBASE [23]. Multiple logistic regression The 10 MTases with significant association with strain origin (Table 1) were used as independent variables for the multiple logistic regression. A logistic regression was calculated to predict the strain origin (Europe versus non-Europe; or Africa versus non-Africa). Considering that the majority of strains are of European origin, the output variable, or dependent variable, was established as Europe/non-Europe. The model was statistically significant (p = 0.00040), Interleukin-2 receptor i.e. the selected independent variables were significant for the output. Four MTases yielded significant results for the logistic regression model: M. AseI, M. FokI,

M. MspI, and M. HpyCH4III. M. AseI expression is associated with the European group and the other 3 MTases with the non-European group (Additional file 2: Table S5). When the dependent variable is Africa/non-Africa origin and we use the same 10 independent variables, the full model is once again significant (p = 0.0001) (Additional file 2: Table S6). For this model we identified 5 significant MTases: M. AseI, M. MspI, M. Hpy188I, M. Hpy99I, and M. HpyCH4III. There was an association of the expression of M. MspI and M. HpyCH4II with African strains (Odds Ratio, OR>1). The other MTases were associated with the strains of non-African origin (OR<1). Multinomial logistic regression A multinomial logistic regression presented a nominal outcome variable with 4 levels: Africa, Asia, America, and Europe.

3 and median value 12.9, range 1.4–75, respectively). Figure 1 Immunohistochemical staining of HIF-1α, LY2603618 mouse VEGF-A and VEGF-C in normal renal tissue (A-C) and clear cell renal cell carcinoma (CCRCC) (D-F). A homogeneous cytoplasmic staining of tubular cells and weak staining in glomerules was observed with HIF-1α (A), while VEGF-A and VEGF-C were positive in tubular cells, glomerular mesangium and interstitial macrophages (B and C). In CCRCC, HIF-1α immmunoreactivity buy AZD0156 was nuclear and/or cytoplasmic (D), while it was perimembranous and/or diffuse cytoplasmic for VEGF-A and VEFG-C (E and F). (magnification ×200). VEGF-A and C Immunohistochemical staining of VEGF-A was cytoplasmic, both in normal renal tissue and tumor cells, as

we described previously [15]. Immunohistochemical staining of VEGF-C was also cytoplasmic in normal renal tissue and CCRCC showing heterogeneous staining of different intensity and percentage of positive Apoptosis Compound Library tumor cytoplasm as well as perimembranous and/or diffuse staining pattern (Fig. 1). Division according to percentage of perimembranous or diffuse staining pattern turned out to be more important than intensity and/or percentage of positive

tumor cytoplasm in relation to HIF-1α or clinicopathologic parameters. The median value of perimembranous staining pattern was 12.7% (range 0–94%) for VEGF-A (pVEGF-A) and 46% (range 0–100%) for VEGF-C (pVEGF-C). The median value of diffuse cytoplasmic pattern was 10% (range 0–92%) for VEGF-A (dVEGF-A) and 26.3% (range 0–100%) for VEGF-C (dVEGF-C). Association between HIF-1α, VEGF-A and -C Nuclear HIF-1α demonstrated inverse correlation with dVEGF-A (p = 0.002) and almost so with dVEGF-C (p = 0.053), and showed no association with perimembranous

staining pattern of either VEGF-A or -C. Cytoplasmic HIF-1α correlated with both dVEGF-A (p < 0.001) and dVEGF-C (p = <0.001), and also showed inverse correlation with perimembranous staining pattern of VEGF-C (p < 0.001), but not VEGF-A (Table 1). Table 1 Relation of HIF-1α to VEGF-A and VEGF-C     VEGF-A (%) VEGF-C (%)     pVEGF-A dVEGF-A pVEGF-C dVEGF-C     p1 rp 1 p1 rp 1 p1 rp 1 p1 rp 1 HIF-1α (%) nHIF-1α 0.535 0.068 0.002 -0.322 0.121 0.168 0.053 -0.209   cHIF-1α 0.094 -0.180 <0.001 0.526 <0.001 -0.629 <0.001 0.637 1Pearson's correlation Regarding association of VEGF-A and -C, Pearson's correlation showed a relation of only diffuse staining pattern of both proteins Sucrase (p < 0.001, rp = 0.586) with no association between the perimembranous staining patterns of the mentioned growth factors. Association of HIF-1α, VEGF-A and -C with clinicopathologic parameters There were 59 men and 35 women in the study. The median value of tumor size was 6.3 (1.8–17.5) cm. The Fuhrman nuclear grading distribution was as follows: 12 (12.8%) grade 1, 40 (42.6%) grade 2, 22 (23.4%) grade 3 and 20 (21.2%) grade 4 tumors. There were 71 (75.5%) tumors limited to the kidney (pT1 and pT2) and 23 (24.5%) tumors with extrarenal expansion (pT3 and pT4).

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women colonized by these species. J Infect Dis 1999, 180:1950–6.CrossRefPubMed 30. Antonio MA, Rabe LK, Hillier SL: Colonization of the rectum by Lactobacillus species and decreased risk of bacterial vaginosis. J Infect Dis 2005, 192:394–8.CrossRefPubMed 31. Priestley CJ, Jones BM, Dhar J, Goodwin L: What is normal vaginal flora? Genitourin Med 1997, 73:23–8.PubMed 32. Schwebke JR, Morgan SC, Weiss HL: The use of sequential self-obtained vaginal smears for detecting changes in the vaginal flora. Sex Transm Dis 1997, 24:236–9.CrossRefPubMed 33. Zhou X, Brown CJ, Abdo Z, Davis

CC, Hansmann MA, Joyce P, Foster JA, Forney LJ: Differences in the composition of vaginal microbial communities found in healthy Caucasian and black women. ISME J 2007, 1:121–33.CrossRefPubMed 34. Thies FL, König W, König B: Rapid characterization of the normal and disturbed vaginal microbiota by application of 16S rRNA gene terminal RFLP fingerprinting. J Med Microbiol 2007, 56:755–61.CrossRefPubMed 35. Koumans EH, Sternberg M, Bruce C, McQuillan G, Kendrick 3-oxoacyl-(acyl-carrier-protein) reductase J, Sutton M, Markowitz LE: The prevalence of bacterial vaginosis in the United States, 2001–2004; associations with symptoms, sexual behaviors, and reproductive health. Sex Transm Dis 2007, 34:864–9.CrossRefPubMed 36. Baele M, Baele P, Vaneechoutte M, Storms V, Butaye P, Devriese LA, Verschraegen G, Gillis M, Haesebrouck F: Application of tDNA-PCR for the identification of enterococci. J Clin Microbiol 2000, 38:4201–4207.PubMed 37. Baele M, Vaneechoutte M, Verhelst R, Vancanneyt M, Devriese LA, Haesebrouck F: Identification of Lactobacillus species using tDNA-PCR. J Microbiol Methods 2002, 50:263–271.CrossRefPubMed 38. tDNA-PCR Library[http://​users.​ugent.

J Ind Eng Chem 2012, 18:449–455. 10.1016/j.jiec.2011.11.029CrossRef 17. Qiu Y, Chen W, Yang S: Double-layered photoanodes from variable-size anatase TiO 2 nanospindles: a candidate for high-efficiency dye-sensitized solar cells. Angew Chem 2010, 122:3757–3761. 10.1002/ange.200906933CrossRef 18. Lin XP, Song DM, Gu

XQ, Zhao YL, Qiang YH: Synthesis of hollow spherical TiO 2 for dye-sensitized solar cells with enhanced performance. Appl Surf Sci 2012, 263:816–820.CrossRef 19. Kim A-Y, Kang M: High efficiency dye-sensitized solar cells based on multilayer stacked TiO 2 nanoparticle/nanotube A-1210477 purchase photoelectrodes. J Photochem Photobiol A Chem 2012, 233:20–23.CrossRef 20. Bakhshayesh AM, Mohammadia MR, Dadar H, Fray DJ: Improved efficiency of dye-sensitized solar cells aided by corn-like TiO 2 nanowires as the light scattering layer. Electrochim Acta 2013, 90:302–308.CrossRef 21. Ferrari AC, Meyer JC, Scardaci V, Casiraghi C, Lazzeri M, Mauri F, Piscanec S, Jiang D, Novoselov KS, Roth S, Geim AK: Raman spectrum of graphene and graphene layers. Phys Rev Lett 2006, 97:187401.CrossRef 22. Yang N, Zhai J, Wang D, Chen Y, Jiang L: Two-dimensional graphene bridges enhanced photoinduced charge transport in dye-sensitized solar cells. ACS Nano

2010, 4:887–894. 10.1021/nn901660vCrossRef 23. Murayama M, Mori T: Evaluation of treatment effects for high-performance dye-sensitized solar cells using equivalent circuit analysis. Thin Sol Film 2006, 509:123–126. 10.1016/j.tsf.2005.09.145CrossRef XAV-939 supplier Competing interests The authors declare that they have no competing interests. Authors’ contributions LCC wrote the paper and designed the experiments. CHH prepared the samples. PSC, XYZ, and CJH did all the measurements and analyzed the data. All authors read and approved the final manuscript.”
“Background SbQ (a styrylpyridinium salt), similar to surfactants, is an amphiphilic sensitizer of the styrylpyridinium family [1], and it produces a very planar stacked rod-like micelle structure. Such a structure makes it possible to stack the molecules with Thalidomide the hydrophobic regions one above the other, with the aldehyde

and nitrogen-methyl groups alternating, and finally produces an aggregate [2]. SbQ can react with amino groups of Apoptosis inhibitor proteins to improve the protein stabilization [3]. Moreover, it can be dimerized via the [2 + 2]-cycloaddition reaction under ultraviolet (UV) irradiation [4]. According to Tao et al. [5], cross-linking of the hydrophobic core via dimerization reaction of the SbQ molecules induced by UV light ultimately produced cross-linked micelles because of hydrophobic interactions between SbQ molecules. Hence, the cross-linked SbQ-montmorillonite (MMT) has potential applications for hydrophobic drug delivery and can be used as an additive into polymeric composites and improve the stability and mechanical properties of polymers [6–9].

Figure 2 Comparison of phylogenetic trees constructed from core and panCB genes. Maximum-likelihood phylogenetic trees of 16 Rhizobiales constructed using the concatenated nucleic acid sequences of 10 housekeeping genes (a) or panC and panB concatenated genes (b). Bootstrap values are

shown over each branch (based on 100 pseudo-replicates). The panCB genes do not fully complement the growth deficiency of a R. etli CFN42 p42f cured derivative in MM It was reported previously that R. etli CFNX186, a p42f-cured derivative of R. etli CFN42, is unable to grow in MM [18]. To assess if the growth deficiency of strain CFNX186 in MM was due to the absence of the panC and panB genes, plasmid pTV4 (panCB) was introduced into strain CFNX186. The growth of the transconjugant (CFNX186-4) after 15 hours of culture in MM was only 50% that of the WT strain SCH772984 cell line grown under the same conditions (Figure 3a). The growth of ABT-263 nmr CFNX186-4 did not improve even after 72 h in culture (data not shown). Interestingly, strain CFNX186-4 had the same growth rate as strain CFNX186 cultured in MM supplemented with 1 μM calcium pantothenate (Figure 3b). This shows that the growth deficiency of CFNX186 is only partly due to the absence of the panCB genes and indicates that other functions encoded in plasmid

p42f are required for growth in MM. Figure 3 panCB genes do not fully restore the growth deficiency of CFNX186. Growth of R. etli CFN42 JPH203 clinical trial wild-type strain, its p42f-cured derivative CFNX186, CFNX186 complemented with the panCB genes (CFNX186-4) and CFNX186 complemented with a 20 kb EcoRI fragment of plasmid p42f containing the panC, panB, oxyR and katG genes (CFNX186-24) in: (a) minimal medium, (b) minimal medium supplemented with 1 μM pantothenate. Growth curves are the mean of at least three independent experiments; error bars represent standard deviations. Previous studies have demonstrated that the katG gene, which encodes Cytidine deaminase the sole catalase-peroxidase

expressed in free-living growth conditions, is located on plasmid p42f of R. etli CFN42. These studies also revealed that the growth rate of a katG mutant in MM was significantly reduced in comparison with that of the wild-type parental strain [19]. On plasmid p42f katG, as well as its putative transcriptional regulator protein encoded by oxyR, are located 80 bp downstream of the panCB genes. We speculated that introduction of the panCB genes together with the katG and oxyR genes might improve the growth of CFNX186 in MM. To test this hypothesis, we used pCos24, which contains a 20 kb fragment of p42f carrying panCB, katG and oxyR (see Material and Methods). pCos24 was introduced into CFNX186 and the resulting transconjugant (CFNX186-24) grown in MM. Figure 3 shows that after 15 hours of culture there was no significant difference between the growth rate of CFNX186 complemented only with panCB (CFNX183-4), and CFNX186 complemented with cosmid pCos24 (CFNX186-24).

This process was repeated twice to ensure purity. Phage purity was confirmed using PCR assays. Amplification of phage stocks was achieved by modifying previous methods [53]. Briefly, mid-exponential phase PAO1 cultures (100 ml) were infected with purified LES phage (MOI = 0.1), at 37°C for 2 h. Lysed cultures were filter-sterilized. Electron microscopy Phage suspensions (1×109 – 1×1010 p.f.u. ml-1) were concentrated by centrifugation, negatively stained with 2% (w/v) uranyl acetate [54], and examined by transmission electron microscopy (magnification x 200,000). Multiplex PCR to confirm pure phage stocks and lysogens Three primer sets,

LESnest1 F/R, buy Entinostat Clust6nest F/R and 4tot1 F/R (Table 4), for the detection of LES phages 2, 3 and 4 respectively, were combined in a multiplex PCR selleck screening library assay for confirmation of each pure phage stock and each PLPL. Colony or filtered phage suspensions were used as templates in each reaction as described previously [25]. Table 4 Primer sequences Primer Sequence (5′-3′) Amplicon (bp) Cycling conditions Reference Multiplex PCR: LES1nestF tttggtgatgatcggcttagc 289 95°C,

4 min then 30 cycles: 95°C, 30 s; 58°C, 30 s; 72°C, 30 s; final extension step, 72°C, 7 min; [25] LES1nestR tgtggaagcgatcagtct       Clust6nestF ggatcgacgtggcataatctg 410   [25] Clust6nestR acgattctccggcatgcagcg       4tot1F gctcatgagtggctgacaac 105   This study 4tot1R tcttgggcagagaaccattc       Q-PCR: 2pro3F caagccctgtctggattttc 102 95°C, 10 min; then 40 cycles: 95°C, 10 s; 60°C, 15 s; 72°C s. This study 2pro3R gagacaggttgggagggagt       3tot1F cgcaggtaccaccagacttt 122   This study 3tot1R catgtccagcaggttcaaaa       3pro3F gcggatgttctcaaacgaat GF120918 ic50 134   This study 3pro3R cgggagaagcaatgacctac     Casein kinase 1   4tot1F gctcatgagtggctgacaac 105   This study 4tot1R tcttgggcagagaaccattc       4pro3F tcgtgctgtgctgatctttt 172   This study 4pro3R agcagtgccagttgatgttg       Preparation of DIG-labeled probes: φ2intDIGF tgcctatctaacggggttca 1097 95°C, 4 min. 30 cycles: 95°C, 30 s; 55°C, 30 s; 72°C, 1 min s; final extension step, 72°C, 7 min This study φ2intDIGR gaagcaaccgagaagtggag     φ3intDIGF ggatcatgtagcgggaaaga 874 This study φ3intDIGR agaacctggcgaaagtctga     φ4cIDIGF atcgttaattggcacggaat

893 This study φ4cIDIGR acagcaacggatttccactc     tot = to quantify total phage copies; pro = to quantify total phage copies. Quantifying production of each LES phage from LESB58 Replication of each LES phage in response to induction of the lytic cycle was compared using Q-PCR to distinguish and enumerate each specific phage type. LESB58 induction experiments were performed on three separate occasions in the presence and absence of norfloxacin for 30 and 60 min exposure times before the 2 h recovery step. DNA was prepared from each replicate using the Bacterial and Virus DNA extraction kit (QIAGEN) and the automated QIAsymphony machine (QIAGEN; pathogen complex 200 protocol). Q-PCR was performed using six specific primer sets to differentiate between prophage and total copies of each phage.