PCR for VIM, IMP, KPC and NDM-1 genes (self designed, Table 1) wa

PCR for VIM, IMP, KPC and NDM-1 genes (self designed, Table 1) was performed for confirmation. Sequence analysis All isolates found to carry ESBL/ampC or carbapenemase gene were further confirmed by sequencing. Sequencing was performed as per manufacturer’s guidelines in 3130×l genetic analyser (Applied Biosystems, Foster city, California). Further the nucleotide and deduced amino acid sequences were analyzed and compared with sequences available in Gene

bank at the National centre of Biotechnology Information (NCBI) web site (http://​www.​ncbi.​nlm.​nih.​gov/​). Results Gut colonization Entinostat research buy pattern of Enterobacteriaceae and distribution of ESBL and AmpC β -lactamases in healthy low birth weight Neonates (1–60 days) On D1, 65.3% of BAY 80-6946 babies were colonized with Enterobacteriaceae with no significant increase on D60. The predominant flora was E. coli on day 1, 21 and 60 followed by Klebsiella pneumoniae (Table 2). Table 2 Distribution of Enterobacteriaceae and associated ESBL and AmpC β- lactamases in Neonates   Total Day 1 Day 21 Day 60 (N = 75) (N = 75) (N = 75) No. (%) No. (%) No. (%) Babies colonized with a least one species   49 (65.3) 48 (64) 53 (70.6) No of babies colonized with at least one ESBL producing isolate   7/49 (14.3) 13/48 (27.1) 22/53 (41.5)* Total Enterobacteriaceae

strains # 267 79 88 100 E.coli 219 69 (87.3) 67 (76.1) 83 (83) Klebsiella pneumoniae 27 3 (3.8) 13 (14.8) 11 (11) Enterobacter sp 14 2 (2.5) 7 (8) 5 (5) Citrobacter Nintedanib (BIBF 1120) sp 5 4 (5.1) 0 1 (1) Salmonella. Typhi 2 1 (1.3) 1(1.1) check details 0 Total ESBL 55 (20.6) 7 (8.9) 17 (19.3) 31 (31)** Total AmpC (N = 39) 53 (19.9) 16 (20.3) 12 (13.6) 25 (25)*** Co-Production of ESBL and AmpC 30 (11.2) 5 (6.3) 9 (10.2) 16 (16)**** Note: Data represents

Enterobacteriaceae isolates from gut of 75 healthy Low birth weight (LBW) neonates on Day 1, 21, 60 of birth. All Figures in parentheses represent percentages. # Some babies had more than one morphologically and biochemically distinct isolates. *p value 0.005 **p value 0.001 ***p value 0.2 ****p value 0.05 when compared to Day 1. Overall ESBL and AmpC production was 20.6% and 19.9% respectively. The total isolates positive for either AmpC and or ESBL were 29.2% (78/267). The predominant phenotypes were co-producers (30/267, 11.23%), followed by only ESBL (25/267, 9.4%) and AmpC (23/267, 8.6%) isolates. Both no. of babies colonized with at least one ESBL producing isolate and ESBL rate amongst Enterobacteriaceae increased three fold (p value 0.005 and 0.001 respectively) from day 1 to day 60, irrespective of associated AmpC production (Table 2). Characteristics of ESBL and AmpC β – lactamases in Enterobacteriaceae isolates from 27 randomly selected neonates The three stool samples from 27 neonates generated 88 gram negative bacilli which included E.

Journal of Nutrition 1993, 123:1939–1951 PubMed 37 Santos RL, Ts

Journal of Nutrition 1993, 123:1939–1951.PubMed 37. Santos RL, Tsolis RM, Baumler AJ, Adams LG: Pathogenesis of Salmonella-induced enteritis. Brazilian Journal of Medical and Biological Research 2003, 36:3–12.PubMed 38. Peuranen S, Tiihonen K, Apajalahti J, Kettunen A, Saarinen M, Rautonen N: Combination of polydextrose and lactitol affects microbial ecosystem and immune responses in rat gastrointestinal

tract. British Journal of Nutrition 2004, 91:905–914.CrossRefPubMed 39. Poulsen M, Molck AM, Jacobsen BL: Different QNZ molecular weight effects of short- and long-chained fructans on large intestinal physiology and carcinogen-induced aberrant crypt foci in rats. Nutrition and Cancer-An International Journal selleck products 2002, 42:194–205.CrossRef 40. Heegaard PMH, Boeg-Hansen TC: Transferrin and alpha-2-macroglobulin-I are circulating acute phase reactants in the mouse. Marker Proteins in Inflammation (Edited by: Bienvenu, Grimaud, Laurent). Berlin, New York: W. de Gruyter 1986, 275–292. 41. Pepys MB, Baltz M, Gomer K, Davies AJ, Doenhoff M: Serum amyloid P-component is an acute-phase reactant in the mouse. Nature 1979, 278:259–261.CrossRefPubMed 42. Palframan RJ, Gibson GR, Rastall RA: Carbohydrate Preferences of Bifidobacterium Species Isolated from the Human

Gut. Current Issues in Intestinal Microbiology 2003, 4:71–75.PubMed Authors’ contributions All authors were part of a project group, which continuously followed and discussed the progress of the experiments. AP designed and carried out the animal studies, performed the statistical analysis and drafted the manuscript. TRL and HF conceived of the study and participated in its design and coordination as well as in the preparation of the manuscript.

ALP carried out the in vitro fermentation study, PMHH carried out the haptoglobin determination, JBA performed PRKACG the fluorescent tagging of the Salmonella strain, RBS performed the immunocytostaining and flow cytometry, and MP contributed to feed design and statistical analysis. SJL and AO contributed significantly to the interpretation of data and the preparation of the manuscript. All authors read and approved the final manuscript.”
“Background Patients with cystic fibrosis (CF), an autosomal recessively inherited disease caused by a mutation in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene, are particularly susceptible to pulmonary infections with Pseudomonas aeruginosa [1, 2]. Colonization of the airways of CF patients with P. aeruginosa results in higher morbidity and mortality LY2606368 molecular weight because of the faster decline of the lung function, especially from the chronic infection phase onwards [3–5]. Detection of colonization and infection by this pathogen as early as possible enables to postpone the chronic infective stage and eventually to achieve the eradication of P. aeruginosa through early treatment.

Strain construction To construct strain NF33, a 400 bp DNA fragme

Strain construction To construct strain NF33, a 400 bp DNA fragment from the region upstream of B. cereus lysK was amplified by PCR using selleck primers NF36F and NF36R, cut with EcoRI and BamHI, and ligated into

similarly restricted pDG268 [28] to produce the plasmid pBCJ307. pBCJ307 was inserted into the amyE locus of the B. subtilis lysine auxotroph strain 1A765 by double crossover to produce strain NF33. In order to analyze the effect of a reduction of the cellular level of charged tRNALys on expression of a P lysK(T box) lacZ fusion, strain BCJ367 was constructed. Plasmid pBCJ307 was integrated into the B. subtilis chromosome GSK2126458 molecular weight by a double crossover event at the amyE locus to produce strain BCJ363. To place the endogenous lysS gene of B. subtilis under IPTG inducible control, plasmid pMUTIN4 [29] was digested with SalI and BsiWI and eluted from an agarose

gel INK 128 to remove the 2 kb lacZ gene. The ends of the plasmid molecule were blunt ended using Klenow polymerase and religated, resulting in plasmid pMUTINXZ. A 670 bp DNA fragment encoding the end of the yacF gene was amplified with oligonucleotides NF2F and NF2R using B. subtiliis strain 168 chromosomal DNA as a template. This fragment was digested with EcoRI and inserted into the EcoRI site of pMUTINXZ, resulting in plasmid pXZ2. Plasmid pXZ2 was then integrated onto the chromosome of strain BCJ363 by a Campbell from type event generating strain BCJ366 thereby placing expression of the lysS gene under the control of the IPTG inducible Pspac promoter. To effect tight control of the Pspac promoter, replicating plasmid pMap65 [30] that encodes a lacI gene, was transformed into BCJ366 to produce strain BCJ367. Strain NF54 was made to assess whether a B. subtilis strain expressing a T-box regulated lysK gene was viable. A 1.95 kb fragment of the B. cereus chromosome encoding the lysK promoter, leader region and structural gene was

generated by PCR using oligonucleotides NF36F and NF9R. This fragment was digested with EcoRI and cloned into the EcoRI site of plasmid pBCJ102 that has transcriptional terminators flanking the multiple cloning site, to generate plasmid pNF30 [31]. A 2567 bp fragment encoding the lysK promoter, T box element and structural gene flanked by transcriptional terminator sequences was amplified using the pBluescript T7 and M13 reverse primers and plasmid pBCJ102 as template. The ends of this fragment were phosphoryalted using T4 polynucleotide kinase (Promega) and it was then cloned into the EcoRV site of plasmid pDG1730 [32] to produce the plasmid pNF48. Plasmid pNF48 was integrated at the amyE locus of the B. subtilis chromosome by a double crossover event to produce strain NF52.

However, there were no differences in RQ or plasma FFA or TG betw

However, there were no differences in RQ or plasma FFA or TG between the dietary groups. Neither lactate nor glucose contents of plasma were different between the groups, so it is not possible to discuss the changes MK 8931 manufacturer in the use of substrates in energy production, which could Captisol explain the differences in oxygen consumption. On the other hand, in the present study, serum albumin increased

during LPVD by 5.8%. This could partially explain the higher oxygen consumption because serum albumin enables a higher rate of FFA transportation to muscle cells [22]. Metabolic acidosis inhibits albumin synthesis [23], so serum albumin content and SID, which both increased during LPVD, refer together to decreased acidosis. More controlled diet interventions this website should be used in the future to clarify this finding. In an earlier study by Galloway and Maughan [21], oxygen consumption increased because of alkalosis, when the subjects exercised at 70% of VO2max, but there was no difference in RQ. It was discussed that alkalosis would have caused a slight change in the use of substrates, which increased the oxygen consumption, but the change was so small that it could not be seen in RQ. In another study [24], metabolic alkalosis induced by NaHCO3 accelerated the increase of VO2 at the onset of high-intensity exercise (87% of VO2max). However, at a lower intensity (40% of VO2max), the alkalosis

had no effect on the kinetics of breathing and oxygen consumption. Acidosis may, in turn, reduce the capacity of hemoglobin to bind oxygen and may reduce the oxygen content of the blood [25]. After LPVD, the subjects may have had an increased capacity to transport oxygen in the blood, but because of the lack of measurable change in acid–base status besides the minor change in SID, this is speculation.

It may also be that LPVD increased the need for oxygen, and as a consequence, oxidation of all substrates increased during submaximal cycling, which could explain the lack of changes in RQ. These results suggest that the energy expenditure was greater and cycling economy poorer after LPVD. In the present study Interleukin-3 receptor insulin-like growth factor 1 (IGF-1) was not measured but according to our recently collected and unpublished data, serum IGF-1 increased during a 7 d high-protein diet and decreased during a 7 d low-protein vegetarian diet. The difference in IGF-1 could be one reason for the difference in oxygen consumption, since lower serum IGF-1 levels may result in poorer exercise economy [26]. In future studies it would be reasonable to control the energy intake of the diets to minimize the effect of difference in caloric intake on performance. However, the subjects were instructed to eat according to their perceived energy needs and they were free to make their own nutritional choices within the given instructions.

These few examples are all that is currently known about the mole

These few examples are all that is currently known about the molecular mechanisms underlying Brucella adhesion and internalization in eukaryotic cells. HeLa cells have extensively been used as a model to investigate the internalization of brucellae of epithelial cells during the colonization ITF2357 manufacturer of

the susceptible host [9, 10]. Here, we employed this cell line to evaluate the rate of invasion of B. melitensis at different growth phases. Our results indicate that cultures of B. melitensis in the late-log phase of growth were more invasive in non-professional phagocytic cells than cultures at mid-log and stationary growth phases. Using cDNA microarrays, we characterized the transcriptome of the most (late-log) and the least (stationary) invasive growth phases of B. melitensis cultures as a preliminary approach for identifying pathogen candidate genes involved in epithelial cell invasion process. Microarray analysis

revealed a greater number of genes up-regulated in these cultures than in stationary https://www.selleckchem.com/products/GDC-0449.html phase cultures. Consistent with the expected differences due to growth, there was a more active metabolism and invasiveness of cultures in late-log phase than cultures in stationary phase. Given the role that some of these genes have in pathogenesis in other bacterial species, we believe that these data may offer insight into potential growth-phase regulated Brucella virulence genes involved in the initial host:pathogen interactions. Results B. melitensis 16 M at late-log phase of growth were more invasive to epithelial cells than were bacteria at Celecoxib mid-log and stationary growth phases As described in the Methods section, B. melitensis was grown to mid-log growth phase, late-log growth phase, or stationary growth phase. At each of these growth phases, bacteria were enumerated, used to infect a representative epithelial cell

line (HeLa cells), and RNA was extracted and microarrays were performed to identify altered gene expression. Under our experimental conditions, there were 0.5 × 109 CFU/ml (OD = 0.18) at the mid-log growth phase, 2 × 109 CFU/ml (OD = 0.4) at late-log phase, and 5 × 109 CFU/ml (OD = 0.72) at stationary phase (Figure 1A). For invasion experiments, a consistent multiplicity of infection (MOI) factor of 1,000 B. melitensis cells per HeLa cell was used to normalize the number of bacteria used. The average number of intracellular bacteria recovered was 60 CFU at mid-log phase, 130 CFU at late-log phase of growth and 27 CFU at stationary growth phase per 103 cells inoculated (Figure 1B). These values represent the average of three C59 wnt price independent experiments. B. melitensis 16 M cultures grown to late-log phase and then co-incubated with HeLa cells for 30 min were therefore 2.2 (P < 0.05) and 4.8 (P < 0.01) times more invasive than were cultures at mid-log and stationary growth phases.

This ratio was determined against white blood cells in whole bloo

This ratio was determined against white blood cells in whole blood:∼7 x 106 cells/ml. Each whole blood sample was incubated with bacteria for 4 hours at 37°C in 5% CO2 Following incubation, plasma was collected by centrifugation at 2000 x g for 10 min at 4°C. The control plasma was obtained in the same way and treated with 0.033 M potassium-phosphate as a mock exposure.

These plasma samples were used for cytokine measurements. Cytokine immunoassays with protein arrays The measurements Apoptosis inhibitor of cytokines were performed using Zyomyx Protein Profiling Biochips (Hayward, CA). These protein arrays allow the simultaneous quantification of 30 biologically relevant cytokines, as determined by Zyomyx, Inc: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12(p40), IL-12(p40/p70), IL-12(p70), IL-13, IL-15, TNFα, TNFβ, Eotaxin, MCP-1, MCP-3, TRAIL, CD95(sFas), find more MIG, sICAM-1, IP-10, CD23, TGF-β, GM-CSF, GCSF, IFN-γ. Each cytokine assay was optimized for the Zyomyx Protein Profiling Biochip based on many factors including the availability of antibodies and the sensitivity and specificity of antibody-cytokine interactions. Each protein array chip is designed with 6 independent microfluidic channels that allow up to 6 samples to be

loaded into isolated regions of an array. Antibodies specific for 30 analytes were arrayed in each channel, and each antibody was arrayed in redundancy on 5 pillars within the channel. Accordingly, a cytokine measurement represents the average of 5 measurements. All immunoassay steps, including sample loading, washing, and detection, were performed with a fully automated biochip processing station (Zyomyx Assay 1200 workstation). Eight protein array chips were used in these experiments. Two chips were used for generating calibration curves with a calibration standard kit containing 30 analytes (Zyomyx, Inc.). Sample (40 μl) was injected into

each channel of the protein array chips. Standard solutions were applied to two channels of each chip for chip-to-chip normalization. Triplicates of control and pathogen-exposed plasmas were applied randomly to four channels of 6 protein array chips. Protein arrays were scanned at 532 nm with Zyomyx Scanner 100 after immunoassays. Zyomyx Data Reduction software was used for normalization, calculation of calibration curves. Dixon’s Selleck Cobimetinib test was used to remove outliers, and the median feature intensity was background subtracted. Concentrations of cytokines in plasma samples were determined by a four parameter logistic model. Cluster analysis of cytokine data Multiple hierarchical clustering methods were used to group the pathogen exposures based on the multivariate cytokine PF-562271 expression profiles induced in a host infection model system. First, hierarchical agglomerative clustering [20] was applied to group the control and the seven pathogen-exposed samples based on their cytokine concentration profiles.

J Appl Microbiol 2010,109(3):808–817 PubMedCrossRef 49 Olier M,

J Appl Microbiol 2010,109(3):808–817.ACP-196 PubMedCrossRef 49. Olier M, Pierre F, Rousseaux S, Lemaitre JP, Rousset A, Piveteau P, Guzzo J: Expression of truncated Internalin A is involved in impaired internalization of some Listeria monocytogenes isolates carried asymptomatically by humans. Infect Immun 2003,71(3):1217–1224.PubMedCrossRef 50. Kim H, Bhunia AK: SEL, a selective enrichment broth for simultaneous growth of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes. Appl Environ Microbiol 2008,74(15):4853–4866.PubMedCrossRef 51. Walcher G, Stessl B, Wagner M, Eichenseher F, Loessner MJ, Hein I: Evaluation of paramagnetic

beads coated ABT-737 nmr with recombinant Listeria phage endolysine derived cell-wall-binding domain proteins for separation of Listeria monocytogenes from raw milk in combination

with culture-based and real-time polymerase chain reaction based quantification. Foodborne Pathog Dis 2010,7(9):1019–1024.PubMedCrossRef 52. Paoli GC, Kleina LG, Brewster JD: Development of Listeria monocytogenes-specific 4EGI-1 in vitro immunomagnetic beads using a single-chain antibody fragment. Foodborne Pathog Dis 2007,4(1):74–83.PubMedCrossRef 53. Tully E, Hearty S, Leonard P, O’Kennedy R: The development of rapid fluorescence-based immunoassays, using quantum dot-labeled antibodies for the detection of Listeria monocytogenes cell surface proteins. Int J Biol Macromol 2006,39(1–3):127–134.PubMedCrossRef 54. Bueno VF, Banerjee P, Banada PP, de Jose MA, Lemes-Marques EG, Bhunia AK: Characterization of Listeria monocytogenes isolates of food and human origins from Brazil using molecular typing procedures and in vitro cell culture assays. Int J Environ Health Res 2010,20(1):43–59.PubMedCrossRef 55. Jacquet C, Doumith M, Gordon JI, Martin PM, Cossart P, Lecuit M: A molecular marker for evaluating the pathogenic potential of foodborne Listeria monocytogenes. J Infect Dis 2004,189(11):2094–2100.PubMedCrossRef 56. Chen Y,

Ross WH, Whiting RC, Van SA, Nightingale KK, Wiedmann M, Scott VN: Variation in Listeria monocytogenes dose responses in relation to subtypes encoding a full-length or truncated internalin A. Appl Environ Microbiol 2011,77(4):1171–1180.PubMedCrossRef Glycogen branching enzyme 57. Varshney M, Yang LJ, Su XL, Li YB: Magnetic nanoparticle-antibody conjugates for the separation of Escherichia coli O157:H7 in ground beef. J Food Protect 2005,68(9):1804–1811. 58. Foddai A, Elliott CT, Grant IR: Maximizing capture efficiency and specificity of magnetic separation for Mycobacterium avium subsp. paratuberculosis cells. Appl Environ Microbiol 2010,76(22):7550–7558.PubMedCrossRef 59. Snapir YM, Vaisbein E, Nassar F: Low virulence but potentially fatal outcome – Listeria ivanovii. Eur J Intern Med 2006,17(4):286–287.PubMedCrossRef 60.

They have a wurtzite structure and are c-axis-oriented on the see

They have a wurtzite structure and are c-axis-oriented on the seeded FTO thin films, as shown in Figures  2

to 3. Interestingly, the ZnO NWs homoepitaxially form on the seed layer, especially on the free surface of polar c-plane ZnO NPs [42, 43]. Their growth is limited by the mass transport of chemical precursors in solution. Both the structural morphology of the ZnO seed layer and the chemicals used in solution govern the typical structural properties of the ZnO NWs such as their length, diameter, and density. The ZnO NWs are not perfectly aligned vertically (i.e., slightly tilted with respect to the normal to the surface) since the polar c-plane ZnO NPs exhibit a significant mosaicity selleck compound (i.e., the c-plane is slightly tilted with respect to the surface plane). Furthermore, ZnO NWs are twisted to each other since the seed layer does not have any in-plane orientation [50], as expected in polycrystalline thin films, and hence drives their STA-9090 order in-plane orientation by homoepitaxial relationship. Figure 1 FESEM images. (a, c, e, g) 40° tilted view and (b, d, f, h) top view of the (a, b) as-grown bare ZnO NWs, (c, d) as-grown ZnO/CdTe core-shell NW arrays, and ZnO/CdTe core-shell NW arrays annealed at (e, f) 300°C and (g, h) 450°C for 1 h, respectively. Figure 2 XRD patterns, degree of preferred orientation, and texture coefficients. (a) XRD patterns

of the as-grown and annealed ZnO/CdTe core-shell NW arrays at 300°C and 450°C for 1 h. (b) Degree Farnesyltransferase of preferred orientation as well as <531 > and <100 > texture coefficients C531 and C100 as a function of annealing temperature. Figure 3 HRTEM image and Fourier-filtered enhancement. (a) HRTEM image of an as-grown ZnO/CdTe core-shell NW. The insets are Fourier-filtered enhancements along the [100] and [1-10] zone axes of the ZnO NW and CdTe NG, respectively. (b) Fourier-filtered enhancement collected at the ZnO/CdTe

interface, as depicted in the blue rectangular area in (a). Importantly, the CdTe NGs uniformly cover the ZnO NWs from their bottom to their top both for as-grown and annealed ZnO/CdTe core-shell NW arrays. The CdTe shell thickness varies in the range of 50 to 100 nm and is see more typically larger on top of the ZnO NWs than on the vertical sidewalls. This indicates that a larger amount of CdTe is deposited on top of the ZnO NWs. The crystallite size as deduced from the Debye-Scherrer law is instead about 32 nm and thus is smaller than the range of the CdTe shell thickness, showing that several layers of CdTe NGs have been deposited. Basically, it also turns out that some CdTe NGs can cover several ZnO NWs, as depicted in Figure  1. The as-grown CdTe NGs have a zinc-blend structure and are polycrystalline, as shown by the XRD patterns in Figure  2a. No epitaxial relationships are thus expected with ZnO NWs since no strong preferential orientation is revealed.

Table 4 The genotype distribution of nt −443 in the OPN promoter

Table 4 The genotype distribution of nt −443 in the OPN promoter by lung cancer TNM stage   The TNM stages of lung cancer Genotypes I + II III + IV P I + II + III

IV P −443             TT 99 65 1.000 125 39 1.000 CT 72 93 4EGI-1 purchase 0.003 123 42 0.798 CC 6 25 <0.001 11 20 <0.001 Effect of SNPs on bone metastasis As shown in Table 2, there were total 31 patients who had CC genotype at nt −443, among them, 20 cases were at stage IV. Surprisingly, all of these 20 cases were diagnosed with bone metastasis. By compared with TT genotype, it demonstrated that CC genotype at nt-443 might significantly increase the risk of development of bone metastasis (p < 0.01). Associations between genotypes in the OPN promoter region and survival Kaplan-Meier estimates of different genotypes at nt −443 in the OPN promoter were shown in Figure 1. The survival rates for patients with the C/C genotype were significantly lower than the survival rates for patients with the other two genotypes (C/T, T/T), and C/T genotype was also significantly lower than the survival rates for patients with

T/T genotype. There were no significant associations between survival and genotypes at the other sites (nt −156 and nt −66, data not shown). Figure 1 Kaplan-Meier survival is significantly lower in lung cancer patients with the C/C genotype as compared to the other two genotypes at nt −443 in Dinaciclib supplier OPN promoter. Discussion Based on my knowledge, it is first time to report the relationship between OPN polymorphisms and 4��8C bone metastasis among NSCLC patients. Lots of evidence suggests that OPN plays a role in the regulation of tumor metastasis

and that OPN expression is particularly high in metastatic tumors [20–22]. OPN is overexpressed in cancers that have a high propensity for forming bone metastases. In bone metastases, OPN is generally associated with the interface between the carcinoma and the bone surface, and this appears to be related to increased bone resorptive activity by osteoclasts [23]. Moreover, high OPN expression in the primary tumor is associated with early metastasis and poor clinical outcome in human gastric cancer and other cancers [19, 20, 24–27]. A recent study suggested that the OPN promoter was associated with NSCLC [28]. In the present study, we focused on the association of these SNPs with TNM stages of lung cancer, this website especially for bone metastasis. Although the distribution of genotypes in the OPN promoter was not significantly different between lung cancer patients and healthy controls, there were significant differences in the distribution of genotypes (CC) at nt −443 between patients with stage IV and other stage lung cancer (Table 4). The survival rates for patients with the C/C genotype were significantly lower than the survival rates of the other two genotypes (C/T, T/T; Figure 1).