We note, however, that expression PS-341 ic50 of RORγ and Runx1, two factors that are essential for NKT cell differentiation 43, was normal in Bcl11bdp−/− mice, indicating that Bcl11b controls NKT cell development independently of these factors. Our expression profiling analyses suggest that Bcl11b is required to prevent premature and inappropriate expression of many genes specifically expressed in mature CD4+ and/or CD8+ T cells. We speculate that Bcl11b may serve as a timing

factor that holds cells in the immature, DP state until a constellation of factors is in place to support SP differentiation. It is likely that the premature SP gene expression program that is induced in the Bcl11b-deficient DP cells reflects both the direct loss of Bcl11b-dependent repression, and the precocious activity of SP-specific transcription factors (such as Klf2, Zbtb7b, Runx3, and Id2). Therefore, our data suggest that correct regulation of SP cell differentiation

involves mechanisms not only to induce cell-specific gene expression programs, but also to prevent these programs from being inappropriately expressed in immature cells. Mechanisms that prevent early expression of differentiation-associated genes have also been described in other systems. For instance, Polycomb-dependent repression has recently been shown to prevent the premature expression of structural genes in differentiating keratinocytes 44. It is of particular interest that that loss of Bcl11b in DP cells expressing low levels of CD3 results in the induction of genes encoding Zbtb7b and Crizotinib research buy Runx3, which are required for, and strongly upregulated during, CD4 and CD8 SP differentiation programs, respectively 45, 46. We found that Bcl11b bound to sequences in the regulatory regions of these genes, suggesting that Bcl11b directly represses

expression of Zbtb7b and Runx3 in immature T cells. The regulation of Zbtb7b has been intensively investigated in recent Adenosine triphosphate years. Induction of Zbtb7b expression occurs downstream of TCR signaling and requires activation of GATA3 expression 47, whereas Runx3 contributes to Zbtb7b repression in CD8-committed cells 19. The mechanisms that render Zbtb7b silent prior to TCR signaling are less well understood but may in part involve repression by Runx complexes 19. Our present data suggest an essential role for Bcl11b in this early silencing, and thus identify another key player in the regulatory network controlling the dynamic regulation of Zbtb7b during T-cell differentiation. However, our results also raise several questions about how Bcl11b participates in Zbtb7b regulation. It will be important to identify activators responsible for Zbtb7b expression in Bcl11b-deficient DP cells, and determine how Bcl11b antagonizes these activators at the transcriptional level in WT cells.

Autoimmune

thyroiditis, or Graves’ disease, is due to increased infiltration of lymphocytes into the thyroid where they recognize the thyroid stimulating hormone receptor; this leads to autoantibody production, tissue necrosis and loss of thyroid function. The importance of CD40 signalling in Graves’ disease was recognized after the discovery that CD40 is present on thyroid epithelial PF-01367338 solubility dmso cells [54], where it interacts with CD40L (CD154)-expressing autoreactive T cells. In agreement with this observation, blockade of the CD40–CD40L interaction with anti-CD40L antibodies has been shown to prevent experimental thyroiditis [55]. Type 1 diabetes, or insulin-dependent diabetes, is caused by autoreactive T cells that recognize antigens such as insulin and glutamic acid decarboxylase Selleck Proteasome inhibitor on B cells

in the islets of Langerhans. B cells also play important roles in disease pathogenesis, as revealed by B cell-deficient NOD mice [56] and treatment of NOD mice with CD40L antibodies [57]. As the CD40 signal is critical for antibody production and Ig class-switching, depletion of CD40+ B cells, or deletion of endogenous B cells, lowers autoantibody production in these mice and decreases disease severity. In addition to CD40+ B, CD40+ T cells are important in the induction of diabetes in NOD mice [58]. The importance of CD40–CD40L has also been underscored in collagen-induced arthritis (CIA). Treatment of mice with collagen type II and anti-CD40L antibodies blocked joint inflammation, serum antibody titres to collagen, synovial infiltrates and erosion of cartilage and bone [59]. Also, when treated with anti-CD40L antibodies, lupus-prone mice showed reduced glomerulonephritis [60]. Similarly, in an open-label study in SLE patients treated with anti-CD40L, humanized mAb exhibited

disease alleviation, including reduced anti-ds-DNA titres [61,62]. Blockade of CD40–CD40L interaction by anti-CD40L antibodies reduced the incidence and severity of T helper type 1 (Th1)-mediated experimental autoimmune uveoretinitis (EAU) in susceptible B10.RIII mice immunized with autoantigen interphotoreceptor retinoid binding protein (IRBP) in complete Freund’s adjuvant (CFA) [63]. Further analysis revealed that in anti-CD40L Nutlin-3 in vivo antibody-treated mice innate responses to autoantigen IRBP were reduced significantly, but no Th2 dominance was observed [63]. The alleviation of EAE and MS by anti-CD40L therapy [64] further signifies the importance of CD40–CD40L axis in autoimmune diseases (Table 1, Fig. 1c). CD134 (OX40), an inducible T cell co-stimulatory molecule, is one of the most extensively studied members of the TNF superfamily. OX40 expression is activation-induced and, once expressed, OX40 binds OX40L (CD134L) present on a variety of cells [65–67]. OX40 signalling promotes T cell activation, induction of cell survival genes and production of cytokines [68]. OX40 signals play crucial roles in autoimmune and viral diseases, cancer and transplantation [68].

Over the next 3 years she suffered from recurrent sinusitis, otitis media, chest infections (sputum cultures positive

for Moraxella catarrhalis and Haemophilus species) and viral warts. She has a sister with features of DBA – low haemoglobin at 10·4 g/dl, raised mean corpuscular volume (MCV), lymphopenia, elevated fetal haemoglobin (HbF) (3%), high erythrocyte adenosine deaminase (eADA) levels, mildly reduced T cell numbers and slight reduction in proliferative responses to standard mitogens. The sister’s immunoglobulin levels, including functional antibody levels, are normal and she has not required any specific therapy for her anaemia. Investigations in infancy showed a normocytic Romidepsin anaemia, normal serum immunoglobulins [IgG 7·3 g/l (normal range 3·0–10·5), IgA 0·28 g/l (0·1–1·2), IgM 1·07 g/l (0·3–1·5)] and good vaccine responses to conjugated Haemophilus influenzae type b and unconjugated pneumococcal polysaccharide vaccines. By the age of 9, serum IgG levels had dropped to 4·94 g/l (normal range 6·0–13·0). Lymphocyte proliferation responses to phytohaemagglutinin, pokeweed mitogen, OKT3, tetanus, varicella check details and herpes antigens were reduced. Intravenous immunoglobulin (IVIG) replacement therapy was commenced, and stopped after 8 years for reassessment of immune function. Four years later, she had persistent anaemia (Hb 10·0 g/dl, MCV 95·6fl) and low IgG (3·37 g/l), IgA (0·96 g/l) and IgM (0·79 g/l). Bone marrow cytogenetic

studies

were normal, excluding microdeletions in 19q13 and 5q- syndrome. Specific antibody tests showed absent antibodies against measles and reduced tetanus and pneumococcal antibody levels. She was diagnosed to have common variable immunodeficiency as no other causes of low IgG and low levels of specific antibodies were identified. High resolution CT scan chest showed evidence of right middle lobe bronchiectasis and bilateral lower lobe bronchiectasis worse on the left. Intravenous immunoglobulin therapy was recommenced at this stage. Lymphocyte subset analysis showed lymphopenia at 833 × 106/µl (normal range 1500–3500), CD3+ T cells 536 (800–2700), helper CD4+ T cells 291 (400–1700), cytotoxic CD8+ T cells 191 (300–1200), CD19+ B cells 158 (100–600) and CD16+CD56+ Ribonucleotide reductase natural killer cells 32 (90–600). B cell studies showed a reduced class-switched memory B cell subset at 2·5%. Lymphocyte proliferation responses to OKT3, phytohaemagglutinin and pokeweed mitogen remained reduced (see Table 1). Peripheral blood eADA level performed recently was high at 594 (normal range 40–100 u/l), consistent with the diagnosis of DBA. She has remained well on home therapy with weekly subcutaneous immunoglobulin infusions over the last 3 years. Polymerase chain reaction (PCR)-based methods for mutation detection.  Genomic DNA was extracted from the patient’s leucocytes with a commercial DNA purification kit, as per the manufacturer’s instructions.

Judging from these reports,

the neutrophil recruitment essential for the elimination of A. baumannii may be induced by Th1-type immune responses, and these Th1-type cytokines may be secreted by NK1.1+ cells. NKT cells can make both the www.selleckchem.com/autophagy.html Th1-type cytokine IFN-γ and the Th2-type cytokines IL-4 and IL-13. These cells appear to play an important role in allergy, autoimmunity, and tumor control. Moreover, NKT cells play an important protective role in bacterial infection (19, 20). However, Bourgeois et al. reported that NKT cells suppressed neutrophil migration into the lung via Th1-type cytokines IFN-γ and IL-12 (41).It is necessary to clarify whether NK cells or NKT cells are important in the migration check details of neutrophils. IL-17A is thought to participate in host defense against various pathogens and induce the production of TNF-α and CXC chemokines in the lung (42–45). In the present study, the expression level of IL-17A increased in lung tissues at 1 day after inoculation of A. baumannii, and up-regulation of IL-17A was delayed by anti-NK1.1 Ab treatment (data not shown). IL-17A and IL-17F may increase the expression level of neutrophil chemotactic factors, including KC (in mouse), MIP-2 (in mouse

and humans), and IL-8 (in humans) and may be driven by lung epithelial cells (46). Also, the IL-17A-producing cells in bacterially infected lungs appear to be γδT cells rather than CD4+ Th17 cells (47–49). In the present study, γδT cells were detected in the lungs of mice with Acinetobacter pneumonia, and their numbers rapidly L-NAME HCl increased up until Day 3 post-inoculation (data not shown). Thus, γδT cells may be involved in neutrophil recruitment and

may directly or indirectly interact with NK1.1+ cells. The detailed molecular mechanisms underlying the role of γδT cells on Acinetobacter pneumonia remain to be elucidated. In conclusion, the results of the present study show that NK1.1+ cells induce neutrophil recruitment by increasing the expression levels of KC during the early phase of Acinetobacter infection. Further understanding of the molecular mechanisms underlying NK1.1+ cell-mediated immune regulation may lead to improved control of A. baumannii infections. This study was supported in part by a Grant-in-Aid for High Technology Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan. We are grateful to Professor Shin-Ichi Nishikawa for supplying the anti-M-CSF monoclonal antibody, AFS98. The authors who have taken part in this study declare that they do not have anything to disclose regarding funding or conflict of interest with respect to this manuscript. “
“Janus kinase (JAK) inhibitors have been developed as anti-inflammatory agents and have demonstrated clinical efficacy in rheumatoid arthritis (RA). We investigated if JAK-3-selective inhibition alone could disrupt cytokine signalling in rheumatoid synovial fibroblasts.

12Stat1 is one of the seven members of a family of STATs – latent cytoplasmic proteins activated by various stimuli (cytokines and growth factors) and involved in the regulation of cell growth and differentiation, immune response and homeostasis.13 Stimulation with IFN-γ results in the activation of Janus kinases (Jak) 1 and 2. Activated Jaks phosphorylate tyrosine residues on the IFN-γ receptor, which serve as STAT1 docking sites. Following phosphorylation of tyrosine 701 (Y701) Tyrosine Kinase Inhibitor Library STAT1 monomers homodimerize, translocate to the nucleus and activate the transcription of target genes14–16 through binding to γ-activated sequence elements (GAS).17 The promoters of IFN-γ-activated

genes usually contain GAS.13 Two putative GAS sequences have been identified in the GILT promoter at 130 and 510 bp upstream of exon 1 of the GILT gene. There are two naturally occurring forms of STAT1: STAT1α and the alternatively spliced isoform STAT1β. STAT1β lacks the 38 amino acid residues in the C-terminal transcriptional activation domain that can bind the histone acetyltransferases p300/CBP.18,19 STAT1 is primarily activated through phosphorylation at tyrosine 701.20 A secondary,

independent, phosphorylation event occurs at serine 727, which is needed for maximal transcriptional activity.21 In addition to its role in regulating the expression of target genes upon stimulation with IFN, STAT1 has also been shown to play a role in the constitutive expression of certain genes: selleck screening library low Molecular mass Polypeptide 2 (LMP2),22,23 caspases24 and major histocompatibility complex (MHC) class I.25 In this study, we investigated whether STAT1 interacts with the GILT

promoter in the absence of IFN-γ. Our data suggest that the presence of Stat1 in a mouse fibroblast cell line correlates with decreased activity of the GILT promoter and decreased constitutive expression of GILT protein. The DNA affinity precipitation assay (DAPA) showed that STAT1 binds with high specificity to putative GAS motifs in the GILT promoter in the absence of IFN-γ stimulation. We also showed that STAT1 residues Y701 and S727 are not required for constitutive STAT1 Methane monooxygenase binding to the GILT promoter. Therefore, phosphorylation of Y701, thought to be necessary for STAT1 homodimerization, is not required for constitutive binding of STAT1 to the GILT promoter. The absence of C-terminal amino acids from the alternatively spliced form of STAT1β does not prevent the binding of STAT1 to the GILT promoter. The remaining N-terminal portion of STAT1 seems to be crucial for binding of STAT1 to the GILT promoter, independently of IFN-γ stimulation. Our experiments indicate that STAT1 residues 426/427 are required for constitutive interaction of STAT1 with the GILT promoter.

Park and coworkers prepared a DNA vaccine encoding a fusion protein between CRT and Bacillus anthracis protective antigen domain IV and showed that it much enhanced antibody responses to the target Ag (15). Kim and colleagues also Antiinfection Compound Library manufacturer demonstrated that a DNA vaccine encoding CRT linked to the N protein of the SARS-CoV is capable of generating strong N-specific humoral and cellular immunity (16). It should be noted, however, that proteins expressed by DNA vaccines may be retained in the endoplasmic reticulum or Golgi after synthesis, thus limiting their ability to induce an antibody response. Moreover, intracellularly expressed

CRT may not be as efficient as soluble extracellular CRT in exerting APC conditioning and in activating B cells in vivo. Nucleocapsid protein, another major structural protein of SARS-CoV, is capable of eliciting strong humoral and cellular immune response in patients and in experimental animals (2, 8, 27). Unlike the S protein, which contains neutralizing epitopes, the N protein cannot induce neutralizing Abs in vivo because it is located inside the viral particles. On the other hand, the S, M and E proteins of SARS-CoV play synergistic roles in viral infection (2) and

Abs against these viral proteins are thought to have a synergistic effect in combating the infectivity of SARS-CoV. selleck compound Thus, a recombinant fusion polypeptide containing CRT/39–272 and the major B cell epitopes in the S, M and E proteins of SARS-CoV may comprise a more favorable vaccine design. In conclusion, rS450–650-CRT has several advantages over rS450–650, including its immunogenicity, stability in solution and simplicity of production. Given that rCRT/39–272 is able to activate human peripheral blood mononuclear cells in vitro (12), this CRT fragment could

be exploited as a molecular adjuvant in the preparation of SARS-CoV vaccines for humans. This study was supported by grants from the Program for Changjiang Scholars and Innovative Research Team in University (IRT1075), the National Foundation of Natural Science of China (30890142/31070781) and National Key Basic Research Programs (2010CB529102). The authors declare that they have no conflicts of interest. “
“Low-density lipoprotein (LDL) apheresis is an extracorporeal Carbohydrate treatment modality used in high-risk patients when LDL cholesterol levels cannot be reduced adequately with medication. The treatment is highly effective, but could be affected by potential unwanted effects on pro- and anti-inflammatory biomarkers. In this paper, we review the literature regarding the effect of LDL apheresis on pro- and anti-inflammatory biomarkers important in atherosclerosis, also as patients in LDL apheresis have high risk for atherosclerotic complications. We discuss the effect of LDL apheresis on complement, cytokines and finally a group of other selected pro- and anti-inflammatory biomarkers.

Research is required to estimate the prevalence of anxiety disorders including comorbid

depression in CKD and examine their influence on functioning and outcomes. Social support refers to an individuals’ perception of the availability and adequacy of social resources and characteristics of social networks. Access to social support has been consistently linked to improved health outcomes in various chronic diseases including CVD.[28] Cohort studies indicate that higher perceived social support predicts decreased risk of mortality,[10, 11, 29] and higher HRQOL in dialysis populations.[29] However, to our knowledge, there are no comparable prospective data in people with CKD. Limited cross-sectional analyses indicate that social support is positively associated with various domains Bortezomib price of HRQOL. For example, higher perceived social support (Multidimensional Scale of Perceived Social Support) has been associated with better BMS-354825 cost cognitive functioning and emotional well-being (Kidney Disease Quality of Life Short-form) in adults with CKD 4.[25] Further, Porter and colleagues found that higher perceived social support (Interpersonal Support

Evaluation List-16) was related to better mental and physical health (SF-36) in a cohort of African Americans with hypertensive CKD.[30] Religious or spiritual affiliation may also play a role in improving health outcomes via enhanced social networks and social support. For example, people who identify as religious or spiritual are often involved in religious communities and typically report higher perceived social support compared with those not identifying as religious.[31] In dialysis patients,

religiosity and spirituality are associated with less depression, greater social support[32] and appears to be an important determinant of HRQOL.[33] Rebamipide Of note, Spinale and colleagues found that higher levels of spirituality predicted improved survival in dialysis patients, with higher social support appearing to mediate this relationship.[34] While preliminary, these studies indicate that improving social networks and social support may be efficacious in people with CKD. The roles of religious and spiritual affiliation in the health of patients with kidney disease before and after dialysis initiation warrant further exploration. HRQOL describes the subjective assessment of the impact of disease and its treatment across the physical, psychological and social domains of functioning and well-being.[35] HRQOL is a marker of disease burden and may be used to assess treatment effectiveness and predict risk for adverse outcomes. Frequently cited dimensions of HRQOL in CKD include depression, anxiety, reduced social interaction, cognitive dysfunction, pain, sleep disturbance, reduced physical functioning, sexual dysfunction, and a reduced global perception of general health or overall quality of life.

Results showed that after being preincubated with 10 μg/ml gp120JRFLD368R, all CNsera find more lost their reactivity with gp120JRFLD368R (Fig. 2B),

suggesting that the gp120-reactive non-CD4bs antibodies in CNsera were completely adsorbed. None of the CNsera except Serum 13 could reactive with gp120JRFL after adsorbed by 10 μg/ml gp120JRFLD368R (Fig. 2B), indicating that only Serum 13 contained CD4bs-specific antibody. Antibodies to glycans are rare, but a number of glycan-specific mAbs have been isolated from HIV-1-infected individuals and shown to exhibit broadly neutralizing activities. 2G12 is one of the most broadly neutralizing mAbs that recognize the glycan moiety on gp120. We investigated whether 2G12-like antibodies were present NVP-AUY922 molecular weight in the sera by analysing their reactivity with gp120IIIB in the presence of D-mannose and showed that the CNsera binding to gp120IIIB was reduced by 15–55% when D-mannose reached 2M (Fig. 3A). As a control, the reactivity of 2G12 to gp120IIIB was completely inhibited by 2M D-mannose, while the reactivities of non-glycan-dependent mAbs (b12, 447-52D) were not affected at all (Fig. 3B), consistent with earlier studies on serum antibodies [31]. Therefore, we conclude that mannose glycan-dependent antibodies widely existed in all eight CNsera. Kifunensine is a mannosidase

inhibitor that Edoxaban blocks Man9GlcNAc2 trimming to Man5GlcNAc2. HIV-1 pseudovirus generated in the presence of kifunensine will carry high mannose glycans [32] and become insensitive to PG9 and PG16 neutralization and more sensitive to 2G12 neutralization [33]. Three pseudoviral isolates (CNE6kifu, CNE55kifu and JRFLkifu) produced in the presence of kifunensine were analysed for their sensitivities to CNsera neutralization. CNE6kifu and CNE55kifu became completely resistant to PG9 neutralization (Fig. 4A), consistent with previous study [33].

Therefore, we used CNE6kifu and CNE55kifu to screen for the PG9-like antibodies in the CNsera. CNE6kifu and JRFLkifu showed higher neutralization sensitivity to 2G12 than CNE6 and JRFL (Fig. 4B). Therefore, CNE6kifu and JRFLkifu were used for probing 2G12-like antibodies in the CNsera. In eight CNsera, only Serum 45 potently neutralized both CNE6 and CNE55, but completely failed to neutralize CNE6kifu and neutralized CNE55kifu with significantly reduced potency (Fig. 5A), suggesting that PG9-like antibodies were present in Serum 45 and contributed a major neutralization activity against these two isolates. N-linked glycosylation at 160 site on virus Env is critical for PG9 recognition and neutralization [11, 33], so we generated an N160K mutant from parental viruses CNE6 and CNE55 and the mutant pseudoviruses CNE6N160K and CNE55N160K were completely resistant to PG9 neutralization (Fig. 4A).

The most important aspect of the study is the effect of the CH-π interaction on TCR recognition of the modified peptide. EGP/Db complexes bind better to the cognate TCRs than complexes with WT peptide, providing a double advantage

of the Pro substitution. To gain insight into this effect, Uchtenhagen et al. used high-powered computers to simulate the simultaneous movements of individual atoms of the structure. Such “molecular dynamics” analysis suggests that increased TCR affinity results from increased rigidity of the peptide within the Db cleft. As with all good science, discoveries beget questions. Most pragmatically, VX-809 cell line can the increased pMHC affinity, pMHC stabilization, and TCR recognition afforded by the p3P substitution be generally

extended to other peptide/MHC combinations for enhanced vaccine efficacy? Previous work by Achour and colleagues this website suggests that p3P altered peptides bind to Db or Kb with increased affinity [23]. Since Y159 is highly conserved among human HLA genes and alleles, this likely applies to human pMHC complexes, particularly for those unusual allomorphs that do not bind with strong p2 anchors (such as B*0801). Can other aromatic residues within the peptide-binding cleft be exploited for CH-π interactions, and if so, will tertiary structure be preserved to maintain TCR recognition? Is increased peptide rigidity generally positive for Anidulafungin (LY303366) TCR recognition? Does increased binding uniformly extend to endogenous peptides when they are loaded on to class I in the ER by the peptide-loading complex? Although binding of exogenous peptides to class I is generally considered to precisely mimic the binding of endogenous peptides, peptides can bind to class II in multiple conformations, depending on how they are loaded, with major biological consequences [26]. The work of Uchtenhagen et al. [18] beautifully illustrates

the importance of continued research on problems thought to be “solved”. It is essential for young scientists in particular to appreciate that nature’s secrets are boundless, and that the critical information for practical applications often springs from surprising sources that are best accessed by curiosity-driven research. This work was supported by the Division of Intramural Research of the National Institutes of Allergy and Infectious Diseases. The authors declare no financial or commercial conflict of interest. “
“One common way to study human leucocytes and cancer cells in an experimental in vivo situation is to use mice that have been genetically engineered to lack an immune system and prevent human cell rejection. These mice lack CD132 and either RAG2 or the catalytic subunit of the DNA-dependent protein kinase, to make the mice deficient in lymphocytes and natural killer cells.

p.m. and (2) stationary. Biofilms were quantified using the standardized crystal violet method (O’Toole check details et al., 1999; Dusane et al., 2008a). Adhesion of bacteria to 96-well polycarbonate microtiter plate surfaces was carried out by inoculating 20 μL of overnight grown culture in 0.5 × LB containing 180 μL of the growth medium. The plates were incubated at 37 °C for 72 h and biofilm formation was estimated by a routine crystal

violet staining method (Dusane et al., 2008b). The experiments were carried out in triplicates. Biofilm formation was also analyzed in glass test tubes (Tomaras et al., 2003). The biofilms were formed by adding 0.1 mL of the culture to 10 mL LB (0.5 ×) dispensed Sirtuin inhibitor in glass test tubes. The experiment was performed in duplicates and

the cultures were incubated at 37 °C for 72 h under two sets of different conditions: (1) shaking at 200 r.p.m. and (2) stationary. After incubation, the medium was removed, the tubes were washed with distilled water, air dried and biofilms were assayed using the crystal violet method. Strains of Escherichia coli HB101 and Pseudomonas aeruginosa PA01 were used as controls for the biofilm experiments (Kazemi-Pour et al., 2007). In vitro assay of bacterial adhesion to the catheter surface was assessed as described earlier with some modifications (Sheth et al., 1983). The selected isolates used for this study were cultivated for 24 h at 30 °C in 0.5 × LB containing 0.25 × minimum

inhibitory concentration (MIC) (0.5 μg mL−1) and 0.5 × MIC (1 μg mL−1) concentrations new of colistin (Sigma, India). After the incubation period, antibiotic was removed from the culture by rinsing twice with sterile saline followed by centrifugation (6000 g for 10 min). The bacterial cells were resuspended in sterile saline and the OD of each suspension was measured colorimetrically at 540 nm to achieve the cell density equivalent to 1–5 × 107 CFU mL−1 (confirmed by plate count). Cultures without antibiotics were used as the controls. Urinary catheters (Rusch GmbH, Kemen, Germany), 7 mm in diameter were cut into 1.5-cm-long segments. The segments were then immersed in 13 × 100 mm tubes containing suspensions of the previously standardized strains and kept at room temperature for 30 min. After this contact, each fragment was placed in a tube (18 × 160 mm) containing 15 mL of sterile saline solution, and the tubes were manually inverted 40 times. This procedure was repeated 15 times, transferring the fragment to 15 tubes successively, with the objective of removing the nonadherent bacteria. After the 15 rinses, the catheter fragments were removed from the tube and rolled over the surface of 10 Petri dishes (90 × 15 mm) containing LB agar. After an incubation period of 24 h at 30 °C, the bacterial colonies were counted.