9 Unfortunately, the measurement of UPC cannot be standardized because urine protein is composed of variable proportions of albumin and other proteins.18 Dip-stick proteinuria correlates poorly with ACR,22,23 while PCR correlates reasonably well with ACR.24 Proteinuria of 0.5 g/day or more usually signifies macroalbuminuria.1,4 However, there have been no studies on the direct comparison between proteinuria and albuminuria in CKD in terms of utilities (biomarker, surrogate end-point and cost-effectiveness). Thus, any comparison between proteinuria and albuminuria in CKD is subject to problems inherent in indirect comparisons.25 Proteinuria and

albuminuria are good biomarkers (Table 1) because they predict clinical end-points (CV events, renal events or mortality) PS 341 in both diabetic and non-diabetic patients.2,3 However, there must be three general lines of evidence to support the acceptance of a biomarker to be a surrogate end-point: biological plausibility, epidemiological data and RCT.3 Despite ample evidence in biological plausibility and epidemiological data, there are limitations in RCT regarding the validity of proteinuria or albuminuria as a surrogate end-point.3 For example, a secondary analysis (but not a primary analysis) of

the Modification of Diet in Renal Disease (MDRD) study indicated that a low BP target slows the GFR decline only in patients with proteinuria of 3 g/day or more.26 Similarly, a secondary analysis of the Prevention RGFP966 of Renal and Vascular End-stage Disease Intervention Trial (PREVEND-IT) found that BP lowering decreases CV events only

in patients with higher albuminuria levels.27 The Ongoing Telmisartan Alone and in Combination with Ramipril Global End-point Trial (ONTARGET) study even found that combined ACEI and ARB therapies decrease ACR while increasing renal outcome.3 Moreover, there has only been one renoprotective RCT with proteinuria as a treatment target to show that a reduction in proteinuria by titrated ACEI decreases Neratinib renal end-points.28 Unfortunately, there have been no RCT with head-to-head comparisons between proteinuria and albuminuria.2 However, a change between normoalbuminuria and macroalbuminuria may be a surrogate for the development or remission of early diabetic nephropathy (Table 1).3 The remission of nephrotic proteinuria is a surrogate for the remission of GFR decline (Table 1).3 Moreover, ACEI- or ARB-induced change in proteinuria or albuminuria is a surrogate for changes in CKD progression in patients with mild to moderate proteinuria (Table 1).3 A randomized trial comparing screening for proteinuria and albuminuria is the best evidence of cost-effectiveness, but modelling is an alternative.29 However, most modelling approaches estimate effectiveness from traditional RCT, which are designed for testing efficacy and are not suitable for cost-effectiveness studies.

Differences were displayed

in comparisons by the maximum intensity of the ICG-concentrations. The maximum slope to the second maximum was found to be predictive in selected vessel types, and specific changes of the flow curve were found to indicate compromised vascular flow. The FLOW 800 tool applied for ICG angiography has shown to be a quick and reliable method for assessing blood flow in vessels in this study. The dynamic assessment of selleck inhibitor the ICG signal allows reliable identification of microanastomotic complications with the described parameters. © 2013 Wiley Periodicals, Inc. Microsurgery, 2013. “
“Introduction: A major drawback to microvascular free flap breast reconstruction is the length of operation—up to 9 hours or more for bilateral reconstruction. This takes a significant mental and physical toll on the surgical team, producing fatigue that may compromise surgical outcome. To facilitate the operation we have incorporated a period of cold ischemia of the flaps such that members of the surgical team can alternate a brief respite during the operation. Methods:

check details We retrospectively reviewed our series of microvascular free flap breast reconstructions performed over a four-year period in which cold ischemia of the flaps were induced. Results: Seventy patients underwent free flap breast reconstruction with 104 flaps. Mean cold ischemia time for all flaps was 2 hours 36 min. Average rest time per surgeon per case was 35 min. Complications included two total flap losses (1.9%), one partial flap loss (1.0%), one anastomotic thrombosis (1.0%), two hematomas (1.9%), three fat necrosis (2.9%), and two delayed healing (1.9%). Statistical analysis revealed that the probability of complications is inversely related to cold ischemia time (P = 0.0163). Conclusion: Cold ischemia facilitates breast reconstruction by allowing the surgical team to alternate breaks during the operation. This helps reduce surgeon fatigue and is well tolerated by the flap. Thus, we believe that the use of cold ischemia is safe and advantageous in microvascular breast reconstruction. © 2010 Wiley-Liss,

Inc. Microsurgery 30:361–367, 2010. “
“Partial necrosis of skin flaps remains a significant problem in plastic and reconstructive surgery. In this study we attempted to evaluate medroxyprogesterone the effect of bone marrow-derived mononuclear cells (BM-MNCs) transplantation on improvement of skin flap survival in a rat random pattern skin flap model. Thirty Wistar rats were divided into three groups with each consisting of 10 rats. BM-MNCs and the adipose-derived stem cells (ADSCs) were transplanted into the subcutaneous tissue in the area where the flap would be dissected. The flaps were then raised two days after cells transplantation. The animals receiving the preoperative Dulbecco’s Modified Eagle Medium (DMEM) treatment were used as the controls. On the 7th postoperative day, the survival areas of flaps were measured and tissues were collected for examinations.

albicans infection in humans. WT C57BL/6 mice or mice lacking TLR7 or TLR9 were infected i.v. with a low dose (1 × 104 CFU) of C. albicans, a challenge that was found to be sublethal for WT mice

in preliminary experiments. Survival and morbidity were monitored daily. As shown in see more Figure 7A, most of the mice lacking either TLR7 or TLR9 succumbed to infection while all WT mice survived. To ascertain whether increased lethality was associated with a decreased ability of these mice to control in vivo infection, we measured fungal burden in the kidney, the main target of hematogenous C. albicans dissemination, at 5 days after infection with the same C. albicans dose (1 × 104 CFU) used in the lethality experiments. In these experiments, we also tested MyD88−/−, IRF1−/−, and 3d mice in addition of TLR7−/− and TLR9−/− animals. While low CFU numbers were found in kidneys of WT mice, fungal burden was significantly increased in mice lacking

either TLR7 or TLR9 (Fig. 7B). Notably, fungal burden was even higher in 3d find more or IRF1−/− mice compared with TLR7−/− or TLR9−/− mice. Mice lacking MyD88 showed the most severe phenotype of all, with colony counts that were approximately 6 orders of magnitude higher than those of WT controls. Collectively, these data indicated that the TLR7/TLR9/MyD88/IRF1 pathway has a nonredundant role in defenses against C. albicans. Moreover, 3d mice (that are unable to mobilize TLR7/9 and other intracellular TLRs to phagosomes) showed a phenotype

that was similar to that of IRF1−/− mice and intermediary between MyD88−/− (highly susceptible) and TLR9−/− or TLR7−/− (moderately susceptible). Our results, showing an increased susceptibility of TLR9−/− mice to C. albicans infection, were apparently in contrast with those of previous studies showing similar [28, 38] or even decreased [14] susceptibility Adenosine of TLR9−/− mice in comparison with WT animals. We hypothesized that these discrepancies could be related to the fact that the cited studies used a higher (1–2 log) challenge doses than the one we used. Therefore, to test this hypothesis, we challenged TLR7- and TLR9- defective mice with a 20-fold higher C. albicans dose than that previously used in the experiments summarized in Fig. 7. Under these conditions, no differences were found in susceptibility to infection between TLR7-or TLR9-deficient mice and WT controls, as measured by kidney colony counts (Supporting Information Fig. 5). This data indicate that the effects of TLR7 or TLR9 deficiency on the outcome of the infection are critically dependent on the challenge dose. The identification of receptors and signal transduction pathways involved in immune responses to fungi is essential to understand the mechanisms underlying the development of mycoses and to devise alternative strategies to control these difficult to treat infections.

Among the others, IL-1 has been shown to be

a key cytokine in initiating and amplifying the inflammatory responses against H. pylori [37-39]. Very recently, IL-1β present in the gastric mucosa has been shown to play an important role in H. pylori-induced epigenetic changes linking inflammation to carcinogenesis [40]. Finally, H. pylori virulence and IL-1B genes contribute to peptic ulcers and intestinal metaplasia [41]. Elevation of Tregs at the site of infection and H. pylori-specific Tregs in the circulation [20, 21] has been suggested as a mechanism of pathogen persistence, on the assumption that Tregs are differentiated cells with professional suppressive function. In this study we show for the first time that H. pylori interacts with human Tregs indirectly via DCs and modifies their function. Our data show that H. pylori-treated DCs stimulate Treg proliferation, diminish their suppressive find more selleck compound function and that DC-derived IL-1β drives this process. Biopsy data from in-vivo H. pylori-infected antrum corroborated these findings, showing that a significant portion of Tregs found in infected gastric biopsies are actively undergoing mitosis. The persistence of H. pylori in the gastric mucosa may allow continual restimulation of the Treg population. This restimulation may allow for expansion of the Treg population beyond the 3-day peak observed in vitro. In this model it is not the presence of Tregs that promote the

Calpain persistence of infection, but rather the persistence of infection that expands the Treg population in an attempt to limit the damage caused by a prolonged and excessive inflammatory response. Demonstrations that suppressive function of Tregs can be undermined by pathogens have been shown previously in the context of L. major and H. hepaticus infections, limiting inflammation while hindering pathogen clearance [18, 19]. Although pathogens can influence Treg function directly, such as through engagement of TLR-2, -4 and -8 [42-44], we found that H. pylori had no direct effect on Tregs and that the changes induced in Treg behaviour could be explained by cytokine production from DCs. We have found that IL-1β plays a central role in mediating the effects of H. pylori on Tregs. This is of particular interest, as virulent strains of H. pylori expressing cagPAI are associated with elevated levels of IL-1β [13, 45]. As a result, the influence of H. pylori DCs on Tregs may be enhanced by the local microenvironment. In addition, IL-1β has a significant inhibitory effect on gastric acid production [46], which encourages H. pylori colonization to spread and downstream pathological events (gastritis and gastric cancer). As IL-1β appears to have a central role in H. pylori biology and its mechanisms of immune evasion and chronic inflammation, it may be revealing to study the relationship between polymorphisms in IL-1β and interactions between H.

This implies that thymically derived natural Treg cells may also play a role in controlling the overall size of the GC response, or upon systemic TGF-β neutralization, other factors or cytokines may partially compensate leading to nominal induction of iTreg cells. The potential role of IL-10 was also examined by repeated administration of a blocking anti-IL-10R mAb. Mice were injected i.p. on day 0 with 1 mg of anti-IL-10R (1B1.3a) mAb or control rat IgG. Starting Selleck Tanespimycin in the second week, 500 μg of anti-IL-10R mAb

or rat IgG was injected twice weekly and continued until the mice were killed. The SRBC were given i.p. on day 0. Similar to anti-TGF-β-treated mice, blockade of the IL-10R resulted in an inability to control the balance of IgM+ to switched GC B cells in the spleen. Although not evident at days 8 and 12, this imbalance became marked at days 18 and 24 and reflected a significant increase in both the frequency and Dabrafenib in vitro total number of IgM− GC B cells (Fig. 9b). Examination of the frequency and number of total B220+ PNAhi B cells showed little difference between anti-IL-10R mAb and control-treated mice, except at day 24 (Fig. 9a). This is again similar to the result observed after TGF-β neutralization, and may likewise reflect the activity of natural Treg cells or the ability of other cytokines to partially compensate.

Finally, to ensure that anti-IL-10R mAb treatment did not directly modulate responding B cells, the GC population was tested for expression of IL-10R. As shown in the Supplementary material, Fig. S3, no expression above background was detected. A large number of studies have documented the role of Treg cells in controlling antibody responses.16–46 Using either in vivo disruption (anti-GITR mAb) or depletion (anti-CD25 mAb) protocols, investigators have shown that loss of Treg-cell activity results in enhanced humoral

responses to experimental antigens,16–22 pathogens23,24 and auto-antigens.17,25–29 In all of these reports, antibody levels directed against the specific ADAM7 antigen or infectious agent were significantly elevated, including IgG,16–27,29 IgA18,25 and even IgE.19,26 Additional studies examined whether adoptive transfer of polyclonal21,30–32,35,37–40 or TCR transgenic33,34,36,41 Treg cells could dampen antibody responses to defined allo-antigens or auto-antigens. In all cases, the transfer of Treg cells significantly lowered or even eliminated serum antibodies directed against these antigens. As GCs serve as the basis for T-cell-driven humoral responses, the current study examined the behaviour of primary splenic GC reactions induced to a number of antigens in mice treated with an anti-GITR mAb (Figs 1–4). After disruption of Treg-cell activity, total SRBC-induced GC B-cell numbers were increased at all time-points examined (days 8–24). A higher proportion of IgM− switched B cells within the GC compartment largely accounted for this increase.

Plates were then washed four times with PBS containing 0.05% Tween-20. Serum sample were diluted 1:300 in PBS and a threefold dilution series BGB324 supplier was performed. A total of 100 μL per well of the serum dilution was transferred to the LCMV-coated plates. After 1 hour of incubation at room temperature, plates were washed four times, followed by incubation with 100 μL per well of HRP-conjugated goat-anti-mouse IgG (Jackson ImmunoResearch) diluted 1:30 000 in PBS, followed by 1 hour incubation. Thereafter, plates were again washed four times and 50 μL per well of the peroxidase substrate OPD (SIGMA) were applied

and the color reaction stopped after 10 min by adding 100 μL per well of 2 M sulfuric acid. OD was determined at a wavelength of 492 nm. LCMV-specific Ab titers were determined by an endpoint titer 0.1 OD over background. To determine the viral antigen specificity of these Abs, cell lysates of LCMV-infected and noninfected B16 melanoma cells check details were immunoprecipitated with IgG from LCMV immune serum that were bound to protein G-coupled sepharose (GE Healthcare). Samples were separated by 4–12% gradient SDS-PAGE (SERVA) and visualized with rabbit anti-LCMV serum

(1:5000), followed by HRP-conjugated donkey anti-rabbit IgG (Dianova). The ECL plus detection system (GE Healthcare) was applied for visualization. Single-cell suspensions of splenocytes were obtained by mechanical disruption. IFN-γ production of CD8+ T cells was determined by intracellular IFN-γ staining (anti-IFN-γ; clone XMG 1.2, ebioscience) after restimulation of 106 splenocytes with 10−7M LCMV GP33 peptide or LCMV NP396 peptide in the presence of 10 μg/mL Brefeldin A (SIGMA). CTL- and NK-cell activity was determined in a 51Cr-release assay. Target cells were loaded with 51Cr for 2 hours at 37°C and then incubated for 5 hours at 37°C with splenocytes that were previously titrated in a threefold

dilution series. Duplicate wells were assayed DNA ligase for each effector-to-target ratio and percentages of specific lysis were calculated. Data were analyzed using SigmaPlot Version 9.0 software. Significant differences were evaluated with Mann–Whitney U-test using InStat3 software (GraphPad). The authors thank Maike Hofmann for helpful discussions and critical comments on the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft DFG (Pi295/6-1 to H.P. and SFB490 to A.W.). The authors declare no financial or commercial conflict of interest. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.

Moreover, both morphotypes should be examined by strain biotyping methods. Beta-N-hexosaminidase (HexNAcase) activity assessed by the api® ZYM test and on CHROMagar Candida® medium (Becton Dickinson, USA) is also discussed. “
“The mode of inhibitory action of Zataria multiflora Boiss. essential oil (EO) on the fungus, Aspergillus flavus, was studied by colony morphology examination, light microscopy, scanning electron

microscopy (SEM) and transmission electron microscopy (TEM). The EO at concentrations used in this study suppressed the size of the colony as well as sporulation. SEM of mycelia treated with given concentrations of EO showed morphological alterations ranging from loss of turgidity and uniformity of mycelia at low concentrations of EO to evident destruction of the hyphae at higher concentration of EO. Semi-thin Sotrastaurin sections of mycelia exposed to different concentrations of EO were analysed by light microscopy and revealed that the major change at level as low as 50 ppm of EO was limited to vacuolisation of cytoplasm resulting in cell swelling, while at higher concentrations, detachment of the cell membrane from the cell wall, deformation of mycelia and shedding the cytoplasm from the cell were the main alterations. These damages were well documented by TEM, which showed that the main sites

of action of EO selleck chemicals were the plasma membrane and cell wall. In conclusion, morphological and structural changes observed in this study may be one of the mechanisms involved in growth inhibition of the fungi and reducing aflatoxin Niclosamide production. “
“Various studies have documented a shift in species distribution in Candida bloodstream infections (BSI), but there are little data from Southeast Asia. This study was performed to determine the species epidemiology and antifungal susceptibilities of Candida species BSI in Singapore. Candida spp. from BSI were collected from a tertiary and secondary referral hospital, and an obstetrics/paediatric hospital over a 3-year period. The most common isolates were Candida albicans (36%), Candida tropicalis (27%), Candida glabrata

(16%) and Candida parapsilosis (16%). Candida parapsilosis and C. albicans were predominant in the paediatric hospital, and C. albicans and C. tropicalis predominant in the other two institutions. Candida tropicalis temporarily replaced C. albicans as the predominant strain from BSI in 2006. Overall, 87.3% of Candida isolates were susceptible to fluconazole, and 10.4% classified as susceptible-dose-dependent. Fluconazole resistance was detected in C. tropicalis (3.6%), C. parapsilosis (2.1%) and C. glabrata (4.0%). Candida albicans is the predominant species isolated from BSI in Singapore. However, non-albicans species accounted for nearly two-thirds of all cases of candidaemia and the relative increase in C. tropicalis infections deserves further investigation.

i.) in all experiments], complete medium containing 0.5 μg mL−1 cycloheximide (Sigma-Aldrich),

10 μM INP0010 was added to the cells; in controls, DMSO (Sigma-Aldrich) was used instead of INP0010. Successful infection was confirmed by immunofluorescence staining of C. pneumoniae-infected HEp-2 cells seeded on glass cover slips (12 mm Ø). At indicated time points, the infected cells were fixed in a shell vial in ice-cold methanol for 15 min and subsequently stained using a fluorescein isothiocyanate-conjugated Ivacaftor mw monoclonal antibody specific for Chlamydia lipopolysaccharide (Pathfinder, Bio-Rad Laboratories) according to the manufacturer’s instructions and visualized by immunofluorescence confocal microscopy. In RNA half-life experiments, the infected cells were treated with 10 μg mL−1 rifampicin at 14 h p.i. and were harvested 1 and 2 h after addition of antibiotic. The control sample (designated 0 h) was collected before the addition of the antibiotic before RNA and DNA isolation. During the isolation procedure, the culture medium was removed, and the cells were washed twice with ice-cold phosphate buffered saline and then lysed using the lysis buffer from an Agencourt RNAdvance cell kit (Beckman-Coulter) as described by the manufacturer. RNA isolation was performed using the indicated kit, also according to the instructions of the manufacturer. RNA samples were purified

by ethanol precipitation. The concentrations and quality of all samples were quantified using a Nanodrop ND-1000 spectrophotometer (A260 nm/280 nm and A260 nm/230 nm) and diluted with diethylpyrocarbonate-treated Carteolol HCl https://www.selleckchem.com/products/ly2157299.html water to appropriate concentrations. All RNA samples were stored at −80 °C till use. DNA samples were collected at the same time points as RNA, and the DNeasy tissue protocol was applied to isolate total DNA from cultured cells (Qiagen). DNA samples were further purified by ethanol precipitation. The

amount and purity of DNA samples were quantified as described above. All DNA samples were stored at −20 °C until use. Each experiment was repeated at least two times. RNA was isolated as described above. Briefly, 35 μg of total RNA was separated on a 1.5% formaldehyde : agarose gel. The RNA was transferred to a Hybond-N membrane (Amersham) overnight, and subsequently cross-linked.32P-labeled probes corresponding to the coding sequences of groEL_1 and incB were generated using a Megaprime DNA labeling system (Amersham) as stipulated by the manufacturer (Sheehan et al., 1995). Chlamydia pneumoniae transcripts were monitored by qRT-PCR (iCycler iQ® Real-Time PCR Detection System; Bio-Rad Laboratories), using an iScript one-step RT-PCR kit with SYBR Green (Bio-Rad Laboratories). The oligonucleotide primers used (Table 1) were designed using beacon designer software (v 6.0; Premier Biosoft International, Palo Alto, CA). Before use, each primer set was run through an annealing-gradient step to achieve optimal amplification conditions.

Depletion of HIV-specific CD8+ IL-10+ cells from PBMCs led to upregulation of CD38 on CD14+ monocytes together

with increased IL-6 production, in response to gag stimulation. Increased CD38 expression was positively correlated with the frequency of the IL-10+ population and was also induced by exposure of monocytes to HIV-1 in vitro. Production NSC 683864 price of IL-10 by HIV-specific CD8+ T cells may represent an adaptive regulatory response to monocyte activation during chronic infection. Interleukin-10 (IL-10) plays a critical role in limiting proinflammatory immune responses that might otherwise cause damage to the host. During infection, the timing and cellular source of IL-10 production find more are essential to the balance between successful pathogen clearance by innate and adaptive responses and the prevention of immune pathology. Mistimed or excessive IL-10 production can interfere with elimination or control of various bacteria, viruses, and protozoa [1]. For example, in the murine lymphocytic choriomeningitis virus model, blockade of IL-10 signalling resulted in clearance of a chronic viral infection by host and vaccine-induced cell-mediated immune responses [2, 3]. It was noted nearly two decades

ago that IL-10 is upregulated from an early stage of HIV-1 infection and this was proposed to underlie Th cell dysfunction [4, 5]. More recent studies reporting enhancement of HIV-specific effector T-cell responses following in vitro depletion of virus-specific IL-10-producing ‘suppressor’ cells or antibody-mediated blockade of IL-10

support this notion [6, 7]. However, IL-10 gene transcription is upregulated in multiple cell types in the peripheral blood during chronic HIV-1 infection [7]. Whether the reported immune suppressive effects are limited to a specific cell subset is unresolved [8]. This is of critical importance for the development of new therapeutic interventions aiming to ameliorate CD8+ and CD4+ T-cell dysfunction in chronic viral infections including HIV-1. An additional consideration Rho is that IL-10 induction in HIV-1 infection may protect the host from excessive immune activation, since diverse pathogens that cause chronic infections drive the expansion of IL-10-producing adaptive or induced T regulatory (Treg) cells in the periphery [9-11]. In support of this notion, rapid induction of strong Treg-cell responses, together with TGF-β and IL-10, was observed in primary SIV infection of African green monkeys, which is typically nonpathogenic, while these responses were delayed in pathogenic SIV infection in macaques [12]. Furthermore, the presence of an IL-10 promoter polymorphism conferring increased cytokine expression was associated with delayed CD4+ T-cell decline in HIV-1 infection [13].

, 1999) but may also be suspended in host material as seen in many chronic infections (Burmølle et al., 2010). Microbiologists have up until the last few decades focused and emphasized the planktonic state over the biofilm state. However, the importance of the biofilm mode of growth is becoming increasingly

recognized as improved methods to study sessile bacteria have become available, and hence the subsequent accumulation of evidence for its widespread presence. It has been suggested that bacteria are predominantly growing as sessile communities rather than as single cells (Costerton et al., 1987; Davey & O’Toole, 2000). Sessile growing bacteria are defined as an assemblage of cells embedded ‘in a self-produced polymeric matrix’. This matrix is EGFR inhibitor very important for the properties of the biofilm, because it offers structural stability and increased tolerance to antimicrobials and immune cells (Stoodley Selumetinib concentration et al., 2002; Anderson & O’Toole, 2008; Mulcahy et al., 2008; Ma et al., 2009). To gain further information on this phenomenon, one has to investigate how a biofilm is established and propagated. The most

common method is the continuous-culture once-through flow system using the model organism Pseudomonas aeruginosa. In this system, media are slowly passed over the biofilm-growing bacteria, which have attached to a cover slip on a flow cell. This in vitro process of P. aeruginosa biofilm formation can be divided into at least five stages: in the first stage, planktonic cells reversibly attach to a vacant surface. Irreversible binding follows this attachment and then multiplication into microcolonies. The microcolonies produce an extracellular polymeric matrix, which in turn envelopes the colonies. After a couple of days, the microcolonies Metalloexopeptidase attain tower- or mushroom-like structures measuring up to 50 μm in the flow cell (Costerton et al., 1995; Davey & O’Toole, 2000;

Stoodley et al., 2002). The extracellular matrix contains a mixture of polysaccharides, proteins, and DNA (Wingender et al., 2001; Whitchurch et al., 2002; Costerton et al., 2003). When the biofilm grows to a size not beneficial for bacterial survival and growth (e.g., owing to nutrient limitations), focal areas of the biofilm are sloughed off. It is hypothesized this enables the otherwise sessile biofilm bacteria to spread and colonize new surfaces and biofilms to spread. Hence, it seems that the biofilm lifecycle by P. aeruginosa is a dynamic process capable of renewing itself (Costerton et al., 1995; Davey & O’Toole, 2000; Stoodley et al., 2002). The biofilm lifecycle and the matrix components have preferably been investigated by means of confocal laser scanning microscopy (CLSM). This method has provided valuable insight into the biofilm development; however, the information on the detailed ultrastructure of the biofilm is difficult to image by light microscopes.