We chose to present the marginal effects rather than conditional effects since it cannot be assumed that the latter will select those variables with ecologically meaningful correlations with assemblage structure. Instead, displaying marginal effects allows

a number of candidate explanatory variables to be visualised in relation to the major gradients of assemblage variation. Table 4 Results of redundancy analysis (RDA) forward selection buy Doramapimod to test the effects of environmental variables on ant functional group and termite feeding group structure across habitat types, listing all marginally significant (p < 0.05) environmental variables included in the final models Ants/termites Environmental variables Conditional effects, λ 2 Conditional effects, p Marginal effects, λ 1 Marginal effects, p a. Ants Leaf litter cover 0.11 0.001 0.11 0.001 Logged forest (LF)     0.08 0.007 Old growth forest (OG) 0.09 0.001 0.08 0.003 Slope     0.07 0.006 Forest quality 0.05 0.016 0.06 0.011 Small saplings cover 0.04 0.042 0.06 0.009 Humus depth     0.05 0.018 Bare ground cover     0.04 0.045 Grass cover     0.04 0.042 Leaf litter depth 0.03 0.038     b. Termites Old

growth forest (OG) 0.33   0.33 0.001 Forest quality     0.26 0.001 Tall poles cover     0.16 0.001 Logged forest (LF)     0.15 0.003 Bare ground cover     0.09 0.022 Slope     0.08 0.033 Leaf litter cover     0.07 0.048 Rocks cover 0.06 0.028     Humus depth 0.05 0.04     Conditional effects (λ2) show the variation explained, and associated significance, for each variable as it was included into the model by forward selection. Marginal

effects (λ1) show the variation MK-8931 order explained by a variable and associated significance level (p), when no other variables are included in the model. Significance of each environmental variable was calculated using Monte Carlo permutation tests with 999 random permutations Results Overall occurrence across habitats A total of 4,931 ants and 1,392 termites were sampled across 944 soil pits and 128 dead wood examinations. Ants were found in every quadrat, in 75 % of soil pits and 51 % of dead ZD1839 wood examinations. Termites were found in 71 % of quadrats, 16 % of soil pits and 16 % of dead wood examinations. Ant occurrences were significantly greater in logged forest than in old growth forest (Kruskal–Wallis χ 2  = 10.72, df = 2, p = 0.005; Wilcoxon rank sum OG-LF, W = 134.5, p = 0.002), but not different between other habitats (Wilcoxon rank sum OG-OP, W = 71.0, p = 0.623; LF-OP, W = 202.5, p = 0.067). Termite occurrence was significantly higher in old growth forest than in logged forest or oil palm CRT0066101 supplier plantation (Kruskal–Wallis χ 2  = 17.66, df = 2, p < 0.001; Wilcoxon rank sum OG-LF, W = 465.5, p < 0.001; OG-OP, W = 142.5, p = 0.001). Encounters with ants were approximately three times more frequent than encounters with termites in old growth forest, 10 times more frequent in logged forest, and 25 times more frequent in oil palm plantation.

The tubes with blood were centrifuged, the GSK1838705A plasma separated, and all plasma samples were stored in an upright position at −20 °C pending analysis. The stereoselective bioanalysis of warfarin in plasma was done using a validated high pressure liquid chromatography

(HPLC) coupled to tandem mass spectrometry (MS/MS) method. In brief, 300 μL of acetonitrile containing internal standards (deuterated S- and R-warfarin) was added to 100 μL of plasma. Following protein precipitation and centrifugation, 15 μL of the supernatant was injected onto the HPLC system. MI-503 concentration The latter consisted of a C18 pre-column (5 μm, 4 × 3.0 mm; Phenomenex, Aschaffenburg, Germany), a Reprosil Chiral-NR analytical column (8 μm, 125 × 3.0 mm; Dr. Maisch GmbH, Ammerbruch, Germany), a Waters Alliance 2795 pump, degasser, and autosampler selleckchem (Waters, Eschborn, Germany). The columns were eluted with a mixture of methanol:5 mM ammonium acetate pH 4.0 (70:30 v/v) for 11 min. The MS/MS analysis (Quattro LC, Micromass, Wythenshawe, UK) was performed in the positive ionization mode, and the limit of detection was 20 ng/mL for both analytes. For R-warfarin, the inter-day coefficients of variation (imprecision)

were ≤11.0 %, whereas inter-day inaccuracy ranged between −1.1 and 0.6 %. For S-warfarin, imprecision was ≤10.1 %, whereas inter-day inaccuracy ranged between −2.0 and −0.4 %. Blood samples for the determination of factor VII and INR were collected pre-dose, and 4, 8, 12, 24, 36, 48, 60, 72, 96, 120, and 144 h after dosing with warfarin during both treatment periods in tubes containing citrate as anticoagulant. These samples were put on ice and sent as soon as possible to the local clinical laboratory for analysis. Farnesyltransferase The assay of factor VII was performed by a standard one-stage method on fresh plasma. The results are expressed in percent of the laboratory reference value. The prothrombin

time of each sample was measured using a standard test and then standardized to yield the INR, a fraction that has no unit. In treatment A, blood samples for determination of trough almorexant plasma concentrations were collected pre-dose on days 1–10 and 24 h after the last almorexant dose on day 10 in tubes with EDTA as anticoagulant. Concentrations in plasma were measured using a validated LC–MS/MS assay with a lower limit of quantification of 0.05 ng/mL and imprecision and inaccuracy ≤4.9 and 5.3 %, respectively [14]. 2.4 Pharmacokinetic and Pharmacodynamic Analyses Pharmacokinetic and pharmacodynamic variables were determined by non-compartmental analysis using WinNonlin Professional Version 5.2.1 (Pharsight Corporation, Mountain View, CA, USA).

The mean fluid intake in these Ironman triathletes was 0.79 ± 0.43 L/h.

In a recent study on 100-km ultra-marathoners showing an association IWP-2 cost between fluid intake and limb swelling, the athletes consumed 0.63 ± 0.20 L/h [60]. Obviously, the 100-km ultra-marathoners consumed less fluid and developed an association between fluid intake and limb swelling in contrast to the present Ironman triathletes drinking more fluids without a relationship between fluid consumption and lower leg swelling. The pathogenesis of lower limb swelling in ultra-endurance athletes may involve the nature of exercise debris, the increased permeability of the capillaries allowing leakage of osmotic material, the ingestion of water to restore/maintain osmotic equilibrium, and the role of lymphatic circulation in clearing the oedemata. We assume that we cannot reduce the swelling in lower selleck kinase inhibitor legs in ultra-endurance athletes due to excessive fluid intake. Strengths and limitations of the present study and implications for future research A strength of this study was that anthropometric measurements were performed immediately upon selleck chemicals arrival at the finish line. A limitation of the present study was that by measuring the entire lower

leg volume, or arm volume, we could not precisely quantify nor locate specifically where the changes in volume occurred. An implication for future research would therefore be to measure the volume of hands and feet separately from the arms and the legs using plethysmography. It would as well be useful to have a measurement method that allows us to differentiate the volume changes occurring in a body part into the different body compositions. Bioelectrical impedance analysis [61] for example is a commonly used method for estimating body compositions, although it measures the composition

of the whole body and not just of one body part [62]. However, this methodology may not provide valid estimates of total body water when hydration status is altered [63] since plasma osmolality and sodium concentration should be unchanged [64, 65]. Regarding the studies from Knechtle et al.[9], Milledge click here et al.[2] and Williams et al.[1] describing an increase in the mean leg volume not immediately after the endurance performance but shortly afterwards, it would also be appropriate to take another measurement later on after the race. Concluding that race time in these Ironman triathletes was relatively short to disturb the body fluid homeostasis [1, 2, 6, 66] it would furthermore be reasonable for future studies to perform these measurements during a longer race such as a Triple Iron ultra-triathlon [7]. Furthermore, we were not able to determine the effect that non-steroidal anti-inflammatory drugs (NSAIDs) had on the decrease of the renal function because we did not trace the consumption of NSAIDs.

A second transcript in the direction complementary to the large transcript in the jamaicamide pathway is probably needed to include jamQ, a gene encoding a condensation like protein that is likely involved with the creation of the pyrrolinone ring of the molecule. According to our RT-PCR experiments, the regions between jamQ and the three genes closest upstream

(ORF5 and ORF6, both transposases, and ORF7, a hypothetical protein), are all transcribed. RG7112 molecular weight In addition, the upstream region of jamQ does not appear to serve as a strong promoter in β-galactosidase reporter assays (see below), despite the presence of possible conserved promoter domains (Table 1). From these data, it appears that jamQ could be part of a larger transcript including these transposases. A larger intergenic region (approximately 1070 bp) lies upstream of ORF7, which could contain the TSS and a promoter for this transcript. The reason for including at least one

transposase in the jamQ transcript is unclear, but this may be a way of ensuring transposable elements have remained associated with the cluster so as to facilitate horizontal gene transfer and pathway evolution. The hectochlorin Y-27632 research buy Biosynthetic gene cluster from L. majuscula JHB [39] contains a transposase (hctC) located between two of the initial genes (hctB and hctD) in the pathway, which is also thought to contribute to the plasticity of the cluster. Biosynthetic investigations using Lyngbya majuscula strains have been highly successful in identifying secondary metabolite check details gene clusters, in part because L. majuscula readily incorporates isotopically labeled precursors in feeding studies [5, 6]. However, further experimentation by way of gene knockout or overexpression in L. majuscula is not yet possible because a viable means of genetic transformation has not been developed. Due to this limitation,

we used genetic constructs in E. coli to determine whether the promoters identified in this study, including the primary pathway promoter upstream of the TSS and PtdIns(3,4)P2 those predicted in intergenic regions, were functional. Although some differences exist in the structure of RNAP between the two bacteria [40], promoter structures in cyanobacteria are often compared to consensus sequences in E. coli [22, 41]. Furthermore, a strong E. coli promoter has been shown to function in the cyanobacterium Synechococcus [37] and the psb2 promoter from Microcystis can be used in E. coli to drive β-galactosidase production [42]. The reporter assay proved effective in verifying the promoter identified upstream of the jamaicamide pathway TSS, as well as several internal promoters located at various regions throughout the gene cluster (Figures 4, 5 and 6).

7%). The observation of a high positive correlation between MIC values of FLC and ITC (r = 0.79

for MIC50 and r = 0.71 for MIC90), in this study, suggests that cross-resistance may be occurring. However, no correlation was observed between www.selleckchem.com/products/blasticidin-s-hcl.html MIC values of the azoles and 24-SMTI, indicating lack of possible cross-resistance. The general finding for our Candida spp. isolates was that they were mostly susceptible to AZA and EIL, because the MIC50s were lower than 2 μg.ml-1 for 73% and 88% of the isolates after treatment with AZA and EIL, respectively. Interestingly, some FLC- and ITC-resistant strains were susceptible to 24-SMTI. However, residual growth of Candida after treatment with AZA was similar to that observed for FLC and ITC. No residual growth was observed after treatment with EIL. The fungicidal action of 24-SMTI was more prominent against CNA species than against C. albicans isolates. A concentration of 4.0 μg.ml-1 of 24-SMTI was enough to kill 100% of C. lusitanae, C. zeylanoides, and C. rugosa, and 50% of C. glabrata. In contrast, this same concentration killed only 4.7% and 9.5% of C. albicans Epoxomicin in vivo isolates, considering AZA and EIL respectively. Previous studies have shown that azasterol derivatives have antifungal activity against

a variety of species [7]. 15-azasterol, in concentrations ranging from 0.01 μg.ml-1 to 4.08 μg.ml-1, inhibits the growth of Saccharomyces cerevisae and C. albicans, with a concomitant accumulation of sterol intermediate molecules [20, 21]. The range of MIC and MFC values for 15-azasterol analogues against these fungal species varied from 0.8 to 3.1 μg.ml-1 and 3.1 to 6.3 μg.ml-1, this website respectively [7] and are similar to the values obtained in the present study. Other azasterol derivatives have been shown to inhibit S. cerevisae 24-SMT, leading to the accumulation of zymosterol [22]. Recent work demonstrated that AZA displays antifungal activity against Paracoccidioides brasiliensis [14] and Pneumocystis carinii [13]. Concentrations of 5 μM (2.05 μg.ml-1) inhibited 100% of

the growth in P. brasiliensis, Carnitine dehydrogenase and the treatment of P. carinii with the IC50 of 0.3 μM (0.12 μg.ml-1) led to growth arrest and accumulation of 24-desakyl sterols, indicating an inhibition of 24-SMT [13]. In addition, previous studies have also shown an anti-protozoan activity of AZA and EIL on T. cruzi epimastigotes and intracellular amastigotes [10], L. amazonensis promastigotes and intracellular amastigotes [11, 12], Toxoplasma gondii [23], and Giardia lamblia [24], with MICs in the low μM to sub-μM range. For protozoans, EIL was reported to be more active than AZA. In contrast, we found in this study that AZA was more active than EIL against Candida spp. isolates. Treatment of C. albicans yeasts with AZA and EIL caused dramatic changes in their cellular and sub-cellular structure.

jejuni among predominant C. coli. Finally, the last step was the application of the real-time

PCR assays to detect and quantify C. coli and C. jejuni in complex substrates like feed, environmental samples, and Ganetespib purchase faeces from experimentally as well as naturally infected pigs. The bacterial culture was used as a gold standard for their validation. Results Specificity, sensitivity and linear range of the real-time PCR assays The specificity of each primers-probe set for the detection of C. coli and C. jejuni was tested

against different strains of C. coli (n = 77) and C. jejuni (n = 54), all of which were correctly identified. Moreover, no signal was observed for any of the other Campylobacter species tested as well as for a range of bacteria, which could be present in faecal samples or responsible for diarrhoea in pigs and humans (Table 1). Finally, the specificity of each real-time PCR assay was characterized for samples using the stool-screening learn more strategy described previously by Lagier et al. (2004) [33]. The DNA extracted from the 30 Campylobacter-negative faecal, feed, and environmental samples and examined in duplicate Lepirudin with each real-time PCR assays produced threshold cycle (Ct) values ≥ 42 when 5 μL of extracted DNA was used as the starting template. All samples in which both duplicates had a Ct value below this threshold were regarded as positive. Table 1 List of strains used

for the validation of specificity of Campylobacter coli and Campylobacter jejuni real-time PCR assays Bacterial species (n) Name or origin of strain C. coli real-time PCR identification C. jejuni real-time PCR identification Campylobacter coli (2) CCUG 11283, CIP 7081 Positive Negative C. coli pig isolates (25) Anses, ENVN-INRA Positive Negative C. coli Fedratinib manufacturer poultry isolates (25) Anses, ENVN-INRA Positive Negative C. coli human isolates (25) Anses, CNR-CH Positive Negative Campylobacter jejuni subsp jejuni (3) CCUG 11284, NCTC 11168, NCTC 81176 Negative Positive C. jejuni CIP 103726 Negative Positive C. jejuni poultry isolates (25) Anses, ENVN-INRA Negative Positive C.

Nanotechnology 2007, 18:465503.CrossRef 10. Horcas I, Fernandez R, Gomez-Rodriguez JM, Colchero J, Gomez-Herrero J, Baro AM: WSXM: a software for scanning probe microscopy and a tool for nanotechnology. Rev Sci Instrum 2007, 78:013705.CrossRef check details 11. Roddaro S, Pingue P, Piazza V, Pellegrini V, Beltram F: The optical visibility of graphene: interference colors of ultrathin graphite on SiO 2 . Nano Lett 2007, 7:2707–2710.CrossRef 12. Blake P, Hill EW, Neto AHC, Novoselov KS, Jiang D, Yang R, Booth TJ, Geim AK: Making graphene visible. Appl Phys Lett 2007, 91:063124.CrossRef 13. Castellanos-Gomez A, Navarro-Moratalla E, Mokry G, Quereda J,

Pinilla-Cienfuegos E, Agraït N, van der Zant HSJ, Coronado E, Steele GA, Rubio-Bollinger G: Fast and reliable identification of atomically thin layers of TaSe 2 crystals. Nano Res 2012, 5:550–557.CrossRef 14. Kaplas T, Zolotukhin A, Svirko Y: Thickness determination of graphene on metal EPZ015666 substrate by reflection spectroscopy. Y Opt Exp 2011, 19:17227–17231. Competing interests The authors declare that they have no competing interests. Authors’ contributions

ADP analyzed the samples by AFM and optical microscopy and suggested the study. XS produced the samples. GG developed the theoretical calculations. LF and GG coordinated the investigation. ADP and GG jointly wrote the manuscript. All authors read and approved the final version of the manuscript.”
“Background Second-generation high-temperature superconducting (HTS) coated conductors based on ReBa2Cu3O7 − δ (REBCO, RE = Y, Gd, Sm, etc., rare earths) films are coming into practical applications for motors, fault current limiters, generators, and transformers [1, 2]. High critical current

(I c) is needed for many HTS applications. Apparently, enhancing the thickness of (RE) BCO films is the simplest method. However, there is an obstacle for this way as there is a current density (J c) decreasing phenomenon as films become thicker [3]. Such a falloff of J c is found in ReBa2Cu3O7 − δ films fabricated by different methods, such as pulsed laser deposition [4], hybrid liquid-phase epitaxy [5], Ba-F-based methods [6], and chemical solution deposition by ink-jet printing [7]. Several possible reasons for the so-called ‘thickness effect’ of J c have been advanced. These include a-axis Amisulpride growth, the increase in surface roughness, and porosity. Another reasonable interpretation of the thickness effect of J c has been Cytoskeletal Signaling inhibitor proposed by Foltyn et al. [8]. They attributed this to misfit dislocations near the interface between the superconductor and the substrate. The same research group reported that by inserting several thin CeO2 layers, the thickness effect can be overcome in some extent [9]. The suppressed thickness effect may be due to much more interfaces between the superconductor and the substrate in the multilayer compared with the single layer.

05). BBR increased protein levels of p53 and FOXO3a through p38 MAPK pathway It has reported that p53 cooperated with BBR-induced growth inhibition and apoptosis of NSCLC cells [6]. In this study, we showed that BBR increased FOXO3a, a transcription factor with known tumor suppressor activity [11], protein expression in a dose-dependent manner (Figure 4A). Similar results were obtained with PC9 cells (not shown). Next, we used special inhibitors of p38 MAPK and ERK1/2 to pre-treated A549 cells to examine the role of these kinases in mediating the effect of BBR on induction of p53 and FOXO3a. As shown in Figure 4B, we found that

the inhibitor of p38 MAPK (SB203580) abrogated BBR-induced p53 and FOXO3a protein expression, while the inhibitors of ERK1/2

(PD98059) had no effect (Figure 4D). Similar results were observed using CH5424802 mouse p38 MAPK siRNAs; intriguingly, we found that silencing of p38α (Figure 4C), but not p38β isoforms (not shown), abrogated the effect of BBR on p53 or FOXO3a protein expression. This result suggested that activation of p38α isoform was involved in the BBR-induced p53 and FOXO3a protein BIRB 796 expression; and that activation of ERK1/2 played no role in this process. Figure 4 Berberine increased p53 and FOXO3a protein expression through p38α MAPK pathway. A, A549 cells were exposed to increased concentration of BBR for 24 h. Afterwards, the expression of FOXO3a and p53 protein were detected by Western blot. B-C, A549 cells were treated with SB203580 (10 μM) (B), or p38α, β siRNAs (70 nM each) (C) for 2 h or 30 h before CUDC-907 manufacturer exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p38 α or β isoforms, p53 and FOXO3a protein was detected by Western blot. D, A549 cells were treated with PD98059 (20 μM) for 2 h and 30 before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the expression of p53 and FOXO3a protein was detected by Western blot. The bar graphs

represent the mean ± SD of p53/GAPDH and FOXO3a/GAPDH of three independent experiments. E-F, A549 cells were treated with SB203580 (10 μM) Nitroxoline for 2 h before exposure of the cells to BBR (25 μM) for an additional 24 h. Afterwards, the cells were collected and processed for analysis of cell cycle distribution by Flow cytometry after propidium iodide (PI) staining (E). And the percentages of the cell population in each phase (G0/G1, S and G2/M) of cell cycle were assessed by Multicycle AV DNA Analysis Software. Data are expressed as a percentage of total cells. Values are given as the mean ± SD of relative percentage of cell cycle phases from 3 independent experiments performed in triplicate. In separated experiment, the cell viability was determined using the MTT assay (F). *indicates significant difference as compared to the untreated control group (P < 0.05). **Indicates significant difference from BBR treated alone (P < 0.05). Previously, we showed that BBR induced cell cycle arrest in G0/G1 phase.

The densitometry values are averages from three independent experiments and are expressed as a ratio of CesT/EscJ signals as assayed by Quantity One software. A dependent, match paired student’s t test was used to assess statistical significance between values (denoted by an asterisk). A representative immunoblot from the experiments is shown. (B) Sucrose density gradient fractionation of membrane preparations from the indicated strains. EscJ and intimin are known inner and outer membrane proteins and their immune-detection served to indicate fractions enriched for inner and outer membranes separated upon ultracentrifugation. Note the altered distribution of CesT in the

presence find more of EscU(N262A) and EscU(P263A). Figure 6 EscU or EscU variants from EPEC lysates do not co-purify with immunoprecipitated CesT. Cell lysates were generated from the indicated bacterial strains and exposed to anti-CesT antibodies in a co-immunoprecipitation experiment. The lysate inputs were probed with the indicated antibodies (top panel). Anti-RNA polymerase antibodies were used to detect RNA polymerase amounts within the lysates which are expected to be equivalent. The elution fractions were probed with the indicated antibodies.

tir and cesT null mutants were included as control strains in the experiment. Note that Tir is unstable in the absence of CesT and therefore was not detected in the elution LY333531 fraction. The lane designations apply to all the panels. Taken together, these data indicate that total CesT membrane levels were not statistically different for EscU variant expressing strains, although the nature of CesT association with the inner membrane was altered in the absence Sodium butyrate or with limited EscU auto-cleavage. CesT retained normal effector binding function in the absence of EscU auto-cleavage and EscU did not AZD5363 solubility dmso co-immunoprecipitate with CesT. Discussion The T3SS is one of the most complex secretory systems in prokaryotic biology,

being composed of at least 10 conserved protein components [17]. The YscU/FlhB proteins have been studied in considerable detail, although the phenotypes associated with secretion are highly variable among bacteria and even within the same species [24, 30–32, 49, 50]. The intein-like auto-cleavage mechanism of EscU was previously elucidated through protein crystallography studies. It was proposed that EscU auto-cleavage likely results in an interface for important protein interactions for type III secretion. In this study, we provide evidence to suggest that EscU auto-cleavage supports efficient type III effector translocation. We also observed that the multicargo type III chaperone CesT was less efficiently associated with the inner membrane (Figure 6), which may partly explain the deficiency in type III effector translocation.

In favor for the first explanation accounts that in the clinical setting the difference was consistent over a large number of biopsies.

AZD8931 solubility dmso The evaluation of the optimal number of reference genes needed to obtain reliable data strengthened the observation that the combination of a Menghini NaCl biopsy followed by RNAlater preservation and an RNAeasy mini kit extraction yields optimal RNA quality from canine liver biopsies. The size of the biopsy needle used in this study was based on a previous study on rat liver biopsy techniques, and turned out to be an optimal balance between quantity and quality of the biopsy and the health risks for the animal [12]. This approach of RNA retrieval proved to be a rapid and feasible method for storage for further molecular analysis, and is in agreement with the findings of others for yeast, human renal and uterine myometrial tissues [15–17]. The quality of the obtained RNA in our approach was feasible AZD2171 chemical structure for micro-array analysis, which requires the highest possible RNA quality, preferential a RIN value above 8.0. Unfortunately our results show that optimal RNA stabilization was only achieved with media that were unsuitable for histology or immunohistochemistry. Histology of RNA later treated biopsies, evaluated in HE and reticulin staining turned out to be of insufficient

quality; furthermore, for the antibodies tested either the background staining was too high or central staining appeared very poor. The best fixative for (immuno)histochemistry proved to be 10% neutral buffered formalin. Boonfix fixation gave good morphology and results in routine HE and reticulin staining, but was suboptimal for the tested immunohistochemical staining methods.

RNAlater fixation yielded poor morphology in routine histology and in immunohistochemistry. Most likely, these LY3023414 cell line shortages in morphological evaluation of RNAlater treated specimens were related to insufficient tissue fixation. Boonfix treated specimens generally evoked less intense reactivity immunohistochemically, but as all tested methods were optimized for use in formalin fixed (24 hrs) wedge biopsy specimens, O-methylated flavonoid they might perform better in a study where the protocols are tailor-made to the fixative. Storage in minus 20°C for Boonfix and RNAlater, as required for molecular purposes, significantly worsened tissue morphology. In our experience staining artefacts more frequently occur in small formalin fixed paraffin embedded biopsies. We hypothesized that in the relatively small biopsies overfixation could easily occur. Therefore an effect of the duration of formalin fixation was assessed with subsequent immunohistochemical evaluation of antibodies to proteins at three different (sub)-cellular locations in addition to routine histological staining methods. Differences of the immunohistochemical reactivity for all three antibodies were found between wedge biopsies and the smaller Menghini tissue samples in this study.