Our results indicate that after exposure to both toxic compounds, arcB transcript levels remain unchanged while those of

ompD and ompC are lowered as compared to untreated cells (Figure 3). Therefore, all the evidence indicates that OM permeability is tightly regulated in response to ROS and could represent a novel mechanism of resistance when bacteria are exposed to these toxic compounds. Figure 3 Effect of H 2 O 2 and HOCl on ompW expression. Wild type (14028s) exponentially growing cells were treated with H2O2 (1.5 mM) or NaOCl (530 μM) for 20 min and ompW, ompD, ompC and arcB mRNA levels were measured by qRT-PCR. Control cells received no treatment. 16S rRNA levels were used for normalization. Values represent the average of four independent experiments ± SD. ArcA binds the ompW promoter region In addition to the soxRS and oxyR systems, several studies have provided evidence that the ArcAB S63845 two component system plays an important role in the resistance to ROS induced damage. For example, ArcA is essential for S. Enteritidis and Typhimurium resistance to ROS [24, 27] and E. coli mutant strains of the sensor ArcB and the regulator ArcA, show an increased susceptibility to H2O2[26]. However, neither of these studies identified genes directly regulated by the system under oxidative stress. We recently

demonstrated that ArcA negatively regulates the expression of S. Typhimurium ompD after H2O2 exposure LY2606368 solubility dmso by direct interaction with its promoter region [12]. To determine if ArcA mediates ompW down-regulation in response to H2O2 and HOCl, a search for putative ArcA binding sites at the ompW promoter region was performed using Virtual Footprint Tacrolimus (FK506) 3.0 [41]. The analysis

predicted the presence of three ArcA binding sites (ABS) located at positions −61 to −70 (ABS-1, forward orientation), -230 to −239 (ABS-2) and −286 to −295 (ABS-3, both in reverse orientation) relative to the experimentally determined transcription start site [42]. Comparison with the extended core region 5′-GTTAATTAAATGTTA-3′ described by Evans et al. (2011) further revealed that only ABS-1 presented a high degree of identity (14 out of 15 nucleotides) with the consensus sequence. To confirm or rule out a direct interaction between ArcA and the predicted binding sites, deletions of the promoter region were generated by PCR (schematized in Figure 4B) and used to perform non-radioactive EMSAs with ArcA and phosphorylated ArcA (ArcA-P). The purity of the protein was assessed by PAGE and ArcA was the dominant product. Electrophoretic mobility shift with ArcA-P was only observed when incubated with fragments that included ABS-1 (Figure 4C and D, W1 and W4). No shifts were observed in fragments that include both ABS-2 and ABS-3 (W3, even at three-fold excess) or control fragments that did not include any ABS (W2 and W5).

Furthermore, our data showing that a loss-of-function mutation in gnd (which produces the second enzyme of the PPP pathway, Figure 2) does not suppress sensitivity to CO2 suggests that the production of 6-phosphogluconate, by either Zwf or gluconate kinase, contributes to CO2 sensitivity in msbB Salmonella. MsbB as a virulence factor? Several publications Dinaciclib cost cite MsbB as a virulence factor that is necessary for both septic shock and the ability to invade and persist in mammalian cells [5, 17, 29]. However, owing to the fact that msbB Salmonella were tested under 5% CO2 conditions,

the lack of virulence may be partially or fully due to the inability of msbB Salmonella to grow in the presence of the 5% CO2. Further experimentation with msbB zwf Salmonella will be necessary to determine which virulence defects are attributable to msbB lipid A and those

that arise from sensitivity to 5% CO2. Based upon this study and earlier studies on the sensitivity of zwf mutant to superoxides, zwf may both reduce virulence on one hand, yet potentiate growth under CO2 conditions on the other, further complicating virulence analyses. Conclusion Here, we report new growth defects in msbB Salmonella: sensitivity to gluconate Danusertib ic50 and growth in hypertonic, acidic or 5% CO2 conditions. These characteristics are in addition to the previously reported growth defects in the presence of salt, EGTA, polymyxin, or MacConkey media. Previous studies showing that MsbB is a virulence factor require further evaluation of the role that CO2 sensitivity plays. The potential for cryptic, spontaneous mutations remains a possibility that should be addressed by re-transduction under non-selective conditions followed by plating independently under CO2 and ambient

air. We have created an msbB somA zwf Salmonella strain that is resistant to growth under acidic or 5% CO2 conditions. This strain contains a loss-of-function mutation Thalidomide in zwf, an enzyme in the pentose phosphate pathway that produces CO2 as it converts a 6 carbon sugar to a 5 carbon sugar. The study of the virulence of msbB zwf Salmonella will allow the determination of what types of virulence are attributable to cells having an MsbB lipid A independent of sensitivity to 5% CO2, which is required for in vitro and in vivo virulence assays. Methods Bacterial strains, plasmids, phage and media The bacterial strains and plasmids used in this study are listed in Table 1. The Salmonella msbB insertion/deletion for tetracycline resistance was described by Low et al. [5]. P22 mutant HT105/1int201 (obtained from the Salmonella Genetic Stock Center, Calgary, Canada) was used for Salmonella transductions. Salmonella enterica serovar Typhimurium strains were grown on LB-0 or MSB agar or in LB, LB-0, buffered LB or MSB broth. MSB media consists of LB (Luria-Bertani media, [30]) with no NaCl and supplemented with 2 mM MgSO4 and 2 mM CaCl2. LB-0 is LB media with no NaCl. Buffered LB pH 7.5 and pH 6.

Among these mechanisms, heavy metal efflux systems have been well-studied [12]. The efflux-mediated mechanism is EPZ015938 basically a plasmid-encoded mechanism involving many operons

such as czcD, chrB, nccA and so on, in which toxic ions enter the cell via active transport (an ATPase pump) or diffusion (a chemiosmotic ion or proton pump) [12, 13]. However, this metal resistance ability is a direct response to the metal species concerned, and consequently, a particular organism may directly and/or indirectly rely on several survival strategies [11]. As a result, microorganisms are viewed as tools for the treatment of wastewater in biological processes, which have demonstrated their advantages over physico-chemical processes. Despite the fact that several microorganisms are known to participate in the detoxification process of wastewater systems and successfully used in the production of effluent of high quality [8], the ability of protozoan species in terms of resistance to and the bioremoval of heavy metals have not been fully documented [14–16]. For decades, protozoan species have been reported as biological

indicators of water quality and pollution rather than metal resistant species due to the sensitivity of certain protozoan species to the pollutants such as heavy metals Lazertinib manufacturer [17]. As a dominant form of

life on earth 1.5 billion years ago and having survived to the present day in unicellular form [18, 19], protozoan species have undeniably passed through considerable challenges and evolutionary change and can also possess the potential to resist and remove heavy metals from wastewater. No specific studies have assessed the resistance of Peranema sp., Trachelophyllum sp. and Aspidisca sp. to highly polluted industrial wastewater systems. Due to the fact that the industrial wastewater is one of the major contributing factors of the water source pollution in South Africa, this study therefore aimed firstly at determining the effect of Benzatropine this source of pollution on the growth response of selected protozoan species compared to selected bacterial species, and secondly, comparing the ability of the test isolates to remove heavy metals. This study was conducted in laboratory-scale reactors which operated in batches. Methods Test organisms In this study, three bacterial species – Bacillus licheniformis ATCC12759, Brevibacillus laterosporus ATCC64 and Pseudomonas putida ATCC31483 – were purchased from Quantum Biotechnologies (Strydompark Randburg, South Africa). These bacterial species have been reported for their metal tolerance or removal [20–23] and antibiotic resistance [24].

g. the Trehalose Phosphorylase pathway, for which putative genes have been identified and partially characterized in N. crassa[40] and A. fumigatus[22] and also exist in A. niger (ANI_1_2720024). However, it is possible to generate mutants,

within the homologous Tps/Tpp group, in A. fumigatus and A. nidulans that totally lack trehalose [11, 12]. Therefore, we believe that this is the only active trehalose synthesis pathway in Aspergilli. However, internal trehalose contents may not solely be dependent on the presence and expression of these six genes, as in S. cerevisiae there is a strong linkage between trehalose synthesis and the degrading trehalases [41] as well as evidences of posttranscriptional activation of the genes involved in trehalose metabolism Selleck CP673451 [42, 43]. Besides a putative phosphatase activity, TppB and TppC may have similar biological roles as the yeast proteins Tps3 and Tsl1, which also contain phosphatase domains – in yeasts, deletion of both genes is necessary before some reduction in internal trehalose content can be observed [17]. It is intriguing that tpsB and tppC are linked on the chromosome. We cannot explain why the conidial trehalose content in this double mutant was significantly higher

after 28 days, but based on the expression AZD5582 supplier patterns (see Figure 3), it is possible that the expression of the two genes are regulated by the same factors. In addition to the above-mentioned observations, some conclusions can be drawn from the gene expression data: All identified genes were expressed, indicating that the paralogs are not inactive duplicates. For tpsC and tppB, the expressions were consistently low after 6 h, indicating that the two genes may be regulated by the same mechanism. This assumption is supported by a previous observation using A. oryzae arrays where the tpsC and tppB orthologs were down-regulated in a deletion strain of atfA,

a gene encoding a transcription factor [44]. To our knowledge, two previous studies describing the expression of LY294002 trehalose synthesis genes in A. niger during germination, using microarray technology, or in combination with RNA sequencing, have been published [29, 45]. With the exception that van Leeuwen and co-workers [29] saw a drastic drop after 2 h and then a gradual up-regulation of tpsA and tpsB, those results are in line with our findings. The extensive measurements of internal trehalose indicate that the trehalose contents, for all strains, were low in 5 day old conidia, significantly elevated in 14 day old conidia, and then maintained at the value of 14 days (Figure 7). A plausible hypothesis is that conidia of A. niger reach full maturity, at least in terms of trehalose accumulation, sometime between 5 days and 2 weeks.

Stockholm University, Stockholm Johnson M, Forsman L (1995) Competence strivings

and self-esteem: an experimental study. Pers Individ Differ 19(4):417–430CrossRef Jöreskog K, Sörbom D (1996) Lisrel 8: user’s reference guide. Scientific Software International Inc, Lincolnwood, IL Karatepe OM, Tekinkus M (2006) Selleck Adriamycin The effects of work–family conflict, emotional exhaustion, and intrinsic motivation on job outcomes of front-line employees. Int J Bank Mark 24(3):173–193CrossRef Kelloway EK, Gottlieb BH, Barham L (1999) The source, nature, and direction of work and family conflict: a longitudinal investigation. J Occup Health Psychol 4(4):337–346CrossRef Kline RB (1998) Principles and practice of structural equation modeling. The Guilford Press, New York Lee RT, Ashforth BE (1993) A longitudinal study of burnout among supervisors and managers: comparisons

between the Leiter and Maslach (1988) and Golembiewski et al. (1986) models. Organ Behav Hum Decis Process 54(3):369–398CrossRef Leiter MP, Durup MJ (1996) Work, home, and in-between: a longitudinal study of spillover. J Appl Bhehav Sci 32(1):29–42CrossRef Leineweber C, Baltzer M, Magnusson Hanson LL, Westerlund H (2012) Work–family conflict and health in Swedish working women and men: a 2-year prospective analysis (the SLOSH study). Eur J Public Health 23(4):710–716 Lidwall U (2010) Långtidssjukskrivna. Beskrivande statistik 1999–2009: kön, ålder, arbetsmarknadsstatus, sjukskrivningslängd, och diagnospanorama [Individuals on long-term sickleave. Desriptive statistics 1999–2009: sex, age, working status, duration AZD3965 of sickness absence,

and medical diagnoses]. Socialförsäkringsrapport [Social insurance report] 2010:11 Little T, Card N (2013) Longitudinal Guanylate cyclase 2C structural equation modeling. The Guilford Press, New York City Little TD, Preacher KJ, Selig JP, Card NA (2007) New developments in latent variable panel analyses of longitudinal data. Int J Behav Dev 31:357–365CrossRef Löve J, Grimby-Ekman A, Eklöf M, Hagberg M, Dellve L (2010) “Pushing oneself too hard”: performance-based self-esteem as a predictor of sickness presenteeism among young adult women and men: a cohort study. J Occup Environ Med 52(6):603–609. doi:10.​1097/​JOM.​0b013e3181dce181​ CrossRef Lundberg U, Mårdberg B, Frankenhaeuser M (1994) The total workload of male and female white collar workers as related to age, occupational level, and number of children. Scand J Psychol 35(4):315–327CrossRef Magnusson Hanson L, Theorell T, Oxenstierna G, Hyde M, Westerlund H (2008) Demand, control and social climate as predictors of emotional exhaustion symptoms in working Swedish men and women. Scand J Public Health 36(7):737–743CrossRef Maslach C, Leiter MP (2008) Early predictors of job burnout and engagement. J Appl Psychol 93(3):498–512. doi:10.​1037/​0021-9010.​93.​3.​498 CrossRef Maslach C, Jackson SE, Leiter MP (1996) Maslach burnout inventory manual, 3rd edn.

We also coded

I of TNM stage as 0, II as 1 and III as 2. As shown in Table 2, the 16278 and 16399 alleles were identified as independent predictors for ESCC outcome. Selleck XAV939 The length of survival for patients with the rare allele 16278T genotype was significantly shorter than that for patients with the frequent allele 16278C (relative risk, 3.001; 95% CI, 1.029 – 8.756; p = 0.044) at the 16278 site. The same was seen for the rare allele 16399G genotype when compared with matched alleles 16399A at the 16399 site in ESCC patients (relative risk, 3.483; 95% CI, 1.068 – 11.359; p = 0.039) (Table 2). These data demonstrated the strong prediction power of 16278C/T and 16399A/G on outcome for ESCC patients. Figure 1 Survival curve according to the nucleotide at position (A)

16274, (B) 16278 and (C) 16399 in D-loop of ESCC patients. Table 2 Multivariate analysis of prognostic factors associated with post-operational survival in ESCC patients with Cox proportional hazards model Factors Relative risk 95% C.I. p see more value Stage of tumor 1.328 0.955-1.848 0.092 16274(G/A) 0 0 0.975 16278(C/T) 3.001 1.029-8.756 0.044 16399((A/G) 3.483 1.068-11.359 0.039 Discussion Selected SNPs in the D-loop region have been examined for the ability to predict cancer risk in other types of tumour [11–14]. The present study has extended those much analyses to determine the cancer risk and the post-operational survival-associated germline SNPs in a continuous sequence of mtDNA between nucleotides 16190 and 583 in ESCC

patients. Three SNPs, 16274G/A, 16278C/T and 16399A/G, were identified for their association with post-operational survival at statistically significant levels by the log-rank test. Multivariate survival analysis identified 16278C/T and 16399A/G to be independent prediction markers for ESCC outcome. We suggest for the first time that SNPs in the D-loop is a prognostic factor in ESCC patients. The relative risk (RR) of death in patients was significantly higher (16278C versus 16278T, RR, 3.001; 95% CI, 1.029 – 8.756; p = 0.044. 16399A versus 16399G, RR, 3.483; 95% CI, 1.068 – 11.359; p = 0.039). Nucleotides 16278 and 16399 are located in hypervariable segment 1 (HV1), which is associated with high rates of mutation [16], but the functional significance of these SNPs in HV1 is not known. Minor alleles of 16278T and 16399G are associated with dramatically shorter period of postoperative survival; the survival curve decreased rapidly in patients carrying these alleles (Figure 1). We compared the distribution frequency of these two SNPs between ESCC patients and normal controls; among 60 age-matched controls, only one carried the 16278T allele and none carried the 16399G allele.

It is clear that alternating bright/dark contrast appears in a periodic manner along the axial direction of the wire in BF TEM images (Figure 2a,c,e), which indicates the existence of planar defect structure. The phenomenon is consistent with the previous report that high density of SFs

in <111> -oriented nanowires commonly form perpendicularly to the growth direction [15]. HRTEM images (Figure 2b,d,f) and corresponding SAED patterns were acquired from the bending areas, which present explicit illustrations of the microstructures in these kink areas. BAY 11-7082 mouse The SAED patterns (Figure 2a,c insets) show the crystal structure of InP NWs here being face-centered cubic (zinc blende). In Figure 2b, it is obvious that the NWs grows along <111> directions and the bending angle is consistent with that between (111) and planes, namely, approximately 110°. Since the 111 planes are the faces with lower energy in the face-centered cubic structure, the growth of NWs through 111 planes is energetically

favorable. Figure 2b also reveals a stacking fault, almost transecting the entire nanowire in the kink area. We suppose that the transecting SFs in the kinked area would be beneficial to the change of growth direction. In addition, nanotwins and SFs were also observed in the region close to approximately 110° kink as depicted in Figure 2d, which corresponds to the selected area in Figure 2c. As mentioned in the previous report [16], the bending of nanowires typically associated with a significantly large local strain in which SFs are induced and resulted to releasing the stress. https://www.selleckchem.com/products/gw3965.html It is as well noted that an approximately 110° kink consisted of successive curves is observed in Figure 2e.

Noticeable contrast variations indicated by white arrows in Figure 2e are supposed to be imaging effects which occur when twin boundary relaxations are present, although it should be pointed out that images with similar appearances could result from astigmatism or misalignment [17]. HRTEM image corresponding to the selected area in Figure 2e is presented in Figure 2f. It is obvious that there is large amount of SFs in the region of approximately 110° kink. In this case, we believe that the larger local strain could be introduced by two successive curves in such narrow N-acetylglucosamine-1-phosphate transferase space. It is noted that most SFs in the kinked area run nearly parallel to the growth direction. We suppose that in the kinked area, a large amount of stress is introduced such that the 111 planes nearly parallel to the growth direction can easily glide and could facilitate the formation of SFs, which plays an important role in releasing the stress. In addition, nanotwins marked by TB are observed in the bending area. According to the literature, twin-plane formation in zinc blende crystals requires very little energy [18]. The twins are as expected for bulk zinc blende crystals, which can twin on 111 planes by rotating through 60° about the <111> axis [19].

To obtain a deep insight into the lattice characteristics of the NWs, TEM imaging were performed along the [−110] zone axis (cubic notation). Figure 3a shows the TEM image of a representative NW of the first group (with indium droplet top ends). The regions ‘1’ , ‘2’ , ‘3’ , and ‘4’ indicate the regions where the selected-area electron diffraction (SAED) analysis is performed. Note that region ‘1’ is on the indium droplet end, while

regions ‘2’ , ‘3’ , and ‘4’ are on the NW body. The SAED spectrum measured at ‘1’ , ‘2’ , ‘3’ , and ‘4’ is shown in Figure 3b,c,d,e, respectively. It can be observed that the 3-deazaneplanocin A purchase indium droplet shows poly-crystalline structures (metal) (see Figure 3b), while the NW body just below the indium droplet present zinc blende structure (InSb semiconductor) (see Figure 3c), which is consistent with previous results reported [15–17] for Au or Ag-catalyzed InSb NWs. The SAED pattern from Bafilomycin A1 manufacturer area 3 (Figure 3d) shows two sets of diffraction patterns [18], and both of them are [1–10] zone axis diffraction patterns.

One pattern indexed by 1 presents a relative 70.5° rotation with respect to the other pattern indexed by 2. 1111 coincides with 11-12, and two patterns reveal the same 111 plane class parallel to growth direction of NW. Figure 3f presents the structural diagram of rotation grain boundary. In Figure 3a, the dark contrast area represents the [1–10] orientation indexed by 1, while the bright contrast area represents the [1–10] orientation indexed by 2. The interfaces between bright areas and dark areas indicate the rotation grain boundaries. There are eight rotation grain boundaries in InSb NW as shown in Figure 3a. The SAED pattern from area 4 is shown in Figure 3e, which shows a cubic zinc blende, the same structure as that shown in Figure 3c. The second group

of InSb NWs (without indium droplet top ends) demonstrates the same lattice structure as the first Phosphoprotein phosphatase group InSb NWs with indium droplet top ends (SAED results are shown Additional file 3: Figure S3). Figure 3 TEM image and SAED pattern of an InSb NW. (a) TEM image of an InSb nanowire terminating with a droplet; (b) SAED pattern from the droplet shown in the area 1 of (a); (c) SAED pattern from area 2 shown in (a); (d) SAED pattern from area 3 shown in (a); (e) SAED pattern from area 4 shown in (a). (f) Structural diagram of rotation grain boundary. Figure 4a shows a typical longitudinal 2θ-ω XRD scan measured from InSb ensemble NW sample. The peaks at 23.8° and 76.3° arise from the 111 and 333 reflections of zinc-blende-structured InSb, respectively [12]. All the observed InSb reflections match those of Si (111), indicating the epitaxial growth of InSb NWs facilitate perpendicular to the Si substrate.

J Colloid Interface Sci 2004, 274:89–94.CrossRef 19. Menon NJ: Dynamic specific heat of a supercooled liquid. Chem Phys 1996, 105:5246.

20. Chen F, Shulman J, Xue Y, Chu CW, Nolas GS: Thermal conductivity measurement under hydrostatic pressure using the 3 ω method. Rev Sci Instrum 2004, 75:4578.CrossRef 21. Cahill DG: Thermal conductivity measurement from 30 to 750 K: the 3ω method. Rev Sci Instrum 1990, 61:802.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions RKN and AKR jointly BIBW2992 in vitro did the planning of the experiment, analysis of the data, and writing the manuscript. RKN did the synthesis, characterization, and the measurements. Both authors read and approved the final manuscript.”
“Background The clinical success of orthopedic and dental implants depends on the interaction between the implanted surface and bone tissues and, consequently, their osseointegration

[1]. Titanium implants are used widely in orthopedic surgery and dentistry for their favorable biocompatibility and corrosion resistance [2, 3]. Surface modification of the implanted material is a critical factor for tissue acceptance and cell survival. Among three different crystalline phases of titania (anatase, rutile, and amorphous titania), anatase phase is more favorable for cell adhesion and proliferation due to lower surface contact angles and/or wettability [4]. Several surface modification techniques, Ricolinostat i.e., sol–gel techniques, chemical (alkali/acid) treatment, anodization, plasma spray, hydroxyapatite-coated surface, and self-assembled monolayers, have been developed and are currently used with the

aim of enhancing the bioactivity of pure Ti surface [5–12]. Over the last decade, bisphosphonates (BPs) have attracted increasing attention as a surface modifier for orthopedic and dental implants. Bisphosphonates are stable pyrophosphates that prevent the loss of bone mass and are used widely to treat a range of diseases with excess bone resorption, such as bone metastasis, hypercalcemia of a malignancy, and Paget’s disease [13–16]. In orthopedic implants, the use of BP is expected to promote osteogenesis at the bone tissue/implant interface by inhibiting the activity of osteoclasts. BPs were reported to inhibit the differentiation of the osteoclast precursor and the resorptive Mannose-binding protein-associated serine protease activity of mature osteoclasts [17, 18]. Furthermore, BPs alter the morphology of osteoclasts, such as a lack of ruffled border and disruption of the actin ring, both in vitro and in vivo[19, 20]. García-Moreno et al. reported that BPs enhance the proliferation, differentiation, and bone-forming activity of osteoblasts directly [21]. Recently, pamidronic acid, a nitrogen-containing bisphosphonate, was reported to conjugate the titanium surface and stimulate new bone formations around the implant both in vitro and in vivo, which contribute to the success of the implant technology [22, 23].

41 1 <0.001 Field width + 5.87 1 0.015 Detrivores Ln(abundance) Age of field margin + 8.732 1 0.003 In all cases farm and year of sampling

were included in the random model. The model estimates are represented graphically in Figs. 2 and 3 NR not relevant Fig. 2 Mean number of taxonomic invertebrate groups (±SE) per age of field margin category. Estimated means and standard errors are based on the HGLM model with age as categorical variable. Trend is based on the same model with age as scale variable. Trend is significantly different from zero (Table 1B) Abundance of functional groups In total, 34,038 predator, Selleck LY333531 11,305 herbivore and 10,720 detritivore individuals were caught with the pitfall traps. Predator abundance was significantly affected by the age category of the field margin (Table 1A); the abundance of predators

decreased with increasing age of the margin (Table 1B; Fig. 3). Herbivore abundance was significantly related to vegetation cover in summer, margin width and age category (Table 1A). A positive relationship with the age of the margin was found (Table 1B; Fig. 3). Detritivore abundance was not affected by age category (Table 1A), but a clear positive correlation between age of the margin and detritivore abundance was found (Table 1B; Fig. 3). Fig. 3 Mean number of individuals of predators, herbivores and detritivores (±SE) per age of field margin category. Estimated means and standard errors are based on the HGLM model of the Ln-transformed Selleckchem RXDX-101 Farnesyltransferase abundance data after correcting for other significant factors and with age as categorical variable. Trends are based on the same model with age as scale variable. All trends are significantly different from zero (Table 1B) Field margin variables Several site-specific variables showed significant relationships with the

age of field margins (Table 2): we found a decrease in the number of plant species (t = −5.585, P < 0.001) and in their evenness (t = −2.651, P < 0.001), the latter indicating that the vegetation is moving towards dominance by certain species. The vegetation cover in summer increased (R = 0.521, P < 0.001). No trends could be detected for nutrient richness, vegetation height in summer and winter, and vegetation cover in winter. Table 2 Significant relations between field margin age and site-specific variables; in a few cases, data for certain margins were lacking (number of replicates is given below each average) Variable (unit); transformation, test   Age 1 2 3 4 5 6 7 8 9 10 11 Sign (Back-transformed) averages (Replicates) Plant species (total number); Ln(x + 1), linear regression t = −5.585 − 18.683 15.653 9.137 17.014 11.375 9.781 8.582 5.989 6.937 10.917 11 P < 0.001 (27) (23) (4) (11) (16) (20) (12) (6) (2) (9) (1) Plant species evenness (E var); untransformed, linear regression t = −2.651 − 0.743 0.614 0.428 0.631 0.620 0.574 0.662 0.470 0.490 0.629 0.63 P < 0.