The authors assume that priority should be given to functional ec

The authors assume that priority should be given to functional ecosystems which provide a multitude of ecosystem services and have a high adaptive capacity to environmental change. Applying

different prioritization categories in the model (e.g. also a ClimateWise pritoritization category) the authors recommend using a combination of ecological, socioeconomic indicators and proxies for vulnerability to climate change in the design of future global conservation strategies. buy GW786034 Outlook What are the overarching lessons learnt that could guide the redirection of conservation strategies for forest biodiversity? Are there check details feasible adaptation strategies to safeguard forest biodiversity in the future? The compilation of papers in this issue demonstrates that research on the impacts of climate change Ro-3306 mw on forest biodiversity can increase knowledge via empirical and modeling approaches. However, uncertainties concerning future climatic development and its impacts remain and conservation strategies have to find approaches to cope with those uncertainties and to integrate new knowledge systematically. The generation of diversity on different levels seems to be a key measure for adapting forest ecosystems to climate change. In the face of future uncertainties, conservation strategies should be actively pushed forward

and should also comprise a diversity of actions in adaptive management within the scope of biodiversity conservation objectives. Such strategies could assist in maintaining the capacity for self-organization of forest ecosystems and hence their resilience (Berkes 2007). They can also help to secure a broad range of possible management options for the future. The papers provide insight into regional and local variation in

the impacts of climate change on forest ecosystems and biodiversity, which should be reflected in future conservation strategies and adaptation measures. In addition to site-specific measures on the small-scale, the landscape level has to be taken increasingly into account. This may determine different conservation objectives and measures on an overarching level. One central aspect in this sense Flavopiridol (Alvocidib) is to increase the permeability of the landscape for different organisms through an increase in habitat diversity and less intensive land uses. Furthermore, the papers revealed that the adaptation of forest conservation strategies to climate change poses challenges for knowledge and decision management. Given the expected changes in site conditions, objectives and measures should be periodically evaluated or re-discussed and adjusted to new insights, according to an adaptive management approach. Such evaluations should be based on scientific findings resulting from models or scenario techniques, but also on management experiences and the local ecological knowledge of different actors and practitioners in forest and conservation management.

In contrast, Verdijk et al investigated the impact of


In contrast, Verdijk et al. investigated the impact of

the protein, casein hydrolysate, on muscle hypertrophy in healthy untrained elderly men [34]. Researchers randomly assigned 28 elderly men to consume either a protein supplement or a placebo pre- and post-workout. Subjects performed a 12-week resistance weight-training program requiring weightlifting 3 d .wk-1. selleck inhibitor Baseline and ending measurements were obtained, including strength assessments, CT scans, DXA scans, blood samples, 24-hour urine samples, muscle biopsies, and immunohistochemistry tests. Results indicated no differences in ending measurements between the protein group and placebo group in muscle hypertrophy, strength, or body composition [34], suggesting that for elderly men, intake of 20 g casein hydrolysate before and after resistance training

does not increase muscle hypertrophy or strength. Anlotinib In this study, however, only 20 g of casein was used, and it was divided into two servings. This protocol would not have provided participants with the required 3 g of leucine needed to maximize protein synthesis. Additionally, since casein is slow digesting [44, 45], it may not have been ideal for use in a study of elderly men. Future studies with this population should incorporate whey protein, which is highly bioavailable in an amount that would provide at least 3 g leucine [29, 30]. Studies comparing the effects of supplementation with adequate protein and those with creatine-enhanced protein pre-and post-workout also should be conducted to determine whether creatine is needed to produce the desired outcomes, as has been demonstrated in Selleckchem MLN2238 younger men [33] (See Table 2). The long-term use of whey protein pre- and post- resistance exercise was investigated

by Hulmi et al. [35], by assigning participants to one of three groups:1) 15 g of whey protein before and after resistance exercise, 2) a placebo before and after resistance exercise, or 3) no supplement no participation in weightlifting but continued habitual exercise as they did prior to the study. Participants in the first two groups completed two resistance exercise sessions per week for 21 weeks consisting of both upper Etofibrate and lower body multi-joint lifts. All participants then had biopsies performed on their vastus lateralis. Results indicated that the whey protein group had significantly greater increases than the other groups in vastus lateralis hypertrophy, and greater overall muscle hypertrophy [35]. These findings provide evidence that whey protein supplementation pre- and post-workout is useful in increasing muscle hypertrophy. Andersen et al. examined the effects of a mixed blend of proteins on muscle strength and muscle fiber size [36].

Protein Expr Purif 2013,90(1):40–46 CrossRef 65 Sitkiewicz I, St

Protein Expr Purif 2013,90(1):40–46.CrossRef 65. Sitkiewicz I, Stockbauer KE, Musser JM: Secreted bacterial phospholipase A2 enzymes: better living through phospholipolysis. Trends Microbiol 2007,15(2):63–69.PubMedCrossRef 66. Shi ZY, Wang H, Gu L, Cui ZG, Wu LF, Kan B, Pang B, Wang X, Xu JG, Jing HQ: Pleiotropic effect of tatC mutation on metabolism of pathogen Yersinia enterocolitica. Biomed Environ Sci 2007,20(6):445–449.PubMed 67. Lavander M,

Ericsson SK, Broms JE, Forsberg A: Twin arginine translocation in Yersinia. Adv Exp Med Biol 2007, 603:258–267.PubMedCrossRef 68. Caldelari I, Mann S, Crooks C, Palmer T: The Tat pathway of the plant pathogen Pseudomonas syringae is required for selleck chemical optimal virulence. Mol Plant Microbe Interact 2006,19(2):200–212.PubMedCrossRef 69. Bronstein PA, Marrichi M, Cartinhour S, Schneider DJ, DeLisa MP: Identification of a twin-arginine translocation system

in Pseudomonas syringae pv. tomato DC3000 and STA-9090 its contribution to pathogenicity and fitness. J Bacteriol 2005,187(24):8450–8461.PubMedCrossRef 70. Ochsner UA, Snyder A, Vasil AI, Vasil ML: Effects of the twin-arginine translocase on secretion of virulence factors, stress response, and pathogenesis. Proc Natl Acad Sci USA 2002,99(12):8312–8317.PubMedCrossRef 71. Feltcher ME, Sullivan JT, Braunstein M: Protein export systems of Mycobacterium tuberculosis: novel targets for drug development? Future Microbiol 2010,5(10):1581–1597.PubMedCrossRef 72. McDonough Farnesyltransferase JA, McCann JR, Tekippe EM, Silverman JS, Rigel NW, Braunstein M: Identification S63845 of functional Tat signal sequences in Mycobacterium tuberculosis proteins. J Bacteriol 2008,190(19):6428–6438.PubMedCrossRef 73. McCann JR, McDonough JA, Pavelka MS, Braunstein M: Beta-lactamase can function as a reporter of bacterial protein export during Mycobacterium tuberculosis infection of host cells. Microbiology 2007,153(Pt 10):3350–3359.PubMedCrossRef 74. McDonough JA, Hacker KE, Flores AR, Pavelka MS Jr, Braunstein M: The twin-arginine translocation pathway of Mycobacterium smegmatis is functional and required

for the export of mycobacterial beta-lactamases. J Bacteriol 2005,187(22):7667–7679.PubMedCrossRef 75. Heikkila MP, Honisch U, Wunsch P, Zumft WG: Role of the Tat ransport system in nitrous oxide reductase translocation and cytochrome cd1 biosynthesis in Pseudomonas stutzeri. J Bacteriol 2001,183(5):1663–1671.PubMedCrossRef 76. Stevenson LG, Strisovsky K, Clemmer KM, Bhatt S, Freeman M, Rather PN: Rhomboid protease AarA mediates quorum-sensing in Providencia stuartii by activating TatA of the twin-arginine translocase. Proc Natl Acad Sci USA 2007,104(3):1003–1008.PubMedCrossRef 77. Chen L, Hu B, Qian G, Wang C, Yang W, Han Z, Liu F: Identification and molecular characterization of twin-arginine translocation system (Tat) in Xanthomonas oryzae pv. oryzae strain PXO99. Arch Microbiol 2009,191(2):163–170.PubMedCrossRef 78.

The relative quantification (RQ) values were

The relative quantification (RQ) values were #PF-6463922 research buy randurls[1|1|,|CHEM1|]# calculated by the following formula: RQ = 2− [ΔCT(mutant) − ΔCT(wild type)][39, 40]. Q-PCR analysis was performed in duplicate (technical replicates) on three independent RNA isolations (biological replicates). β-galactosidase assays To study the

mbo operon expression in different genetic backgrounds, the mbo operon promoter (P mboI ) cloned into pMP220 [19] as previously described [6] was used. The derivative mutants in mgoA, gacA and gacS genes were transformed with plasmid pMP::P mboI which contains the P mboI . The plasmid pLac-mgoBCAD (harboring the mgo operon) was also used to complement the mgoA, gacA and gacS mutants and finally the β-galactosidase activity of P mboI was measured. In order to evaluate the effect of the mgo operon on the activity of P mboI , P. protegens Pf-5 was used due to the BIBW2992 in vitro absence of the two operons in its genome. First, P. protegens Pf-5 was transformed with the pMP::P mboI and the promoter activity was measured, and secondly to measure the effect on the mbo operon transcription, this strain containing the plasmid pMP::P mboI , was also transformed with

the plasmid pLac-mgoBCAD (mgo operon under pLac regulation). As a negative control the β-galactosidase activity was measured for the wild type strain P. syringae pv. syringae UMAF0158 and each strain used in this assay, transformed with empty vector pMP220. β-galactosidase activities were quantified by the Miller method [41]. Briefly, an overnight culture obtained as previously described in growth curve and toxins assay section were prepared. The samples were collected at 18 h, and the cells were harvested and suspended in assay buffer to eliminate any error in the detection of β-galactosidase activity due to the effects of different carbon sources present in the growth medium. The results presented are from three separate experiments, each conducted in triplicate. Phylogeny of the mgoA gene In order to identify the presence of the mgoA gene in the different genomes of Pseudomonas

strains, the mgoA gene from P. syringae pv. syringae Aprepitant UMAF0158 was used in BLASTP [42] comparisons with whole genome sequences of Pseudomonas spp. available in the databases. Once the amino acid sequences of all the orthologous mgoA genes were obtained, the putative adenylation domains were identified using the PKS/NRPS Analysis Web-site (http://​nrps.​igs.​umaryland.​edu/​nrps) [43]. Other adenylation domains of which the activated amino acid is already known were obtained from the database and from De Bruijn met al. [44]. Two phylogenetic analyses were done, the first was using the adenylation domain of all the NRPSs (328 residues) and the second was using the almost entire sequence of MgoA (1015 residues).

Final analysis revealed that the addition of bevacizumab to IFL s

Final analysis revealed that the addition of bevacizumab to IFL significantly improved OS (primary endpoint, HR: 0.66, p < 0.001), PFS (HR: 0.54, p < 0.001) and RR (44.8% vs 34.8%, p = 0.004). The planned analysis comparing patients treated with 5-FU/LV plus bevacizumab with those concurrently enrolled in the IFL plus placebo group, revealed no significant differences between arms in terms of OS (HR:

0.82 [0.59-1.15], selleck chemicals llc p = 0.25), PFS (HR: 0.86 [0.60-1.24], p = 0.42) and RR (49% vs 37%, p = 0.66) [3]. The outcome reported in the 5-FU/LV plus bevacizumab arm was consistent with other experiences that explored the use of bevacizumab in combination with 5-FU/LV. In a phase II randomized study, including 104 patients, the combination of bevacizumab with 5-FU/LV resulted in longer time to disease progression (TTP, median TTP: 9.0 months [5.8-10.9] vs 5.2 months [3.5-5.6]) and in better, but not significantly, RR (40% [24-58] vs 17% [7–23]-34) and OS (median OS: 21.5 months [17.3-undetermined] vs 13.8 months [9.1-23]) [4]. Similar results were R788 chemical structure obtained in another phase II trial, randomizing 209 patients, that were not optimal candidates for irinotecan-containing regimens, to receive 5-FU/LV plus or minus bevacizumab. Patients treated with the antiangiogenic obtained a significantly

ABT-888 cost longer PFS (HR: 0.50 [0.34-0.73], p = 0.0002) and OS, that was the primary endpoint of the study (HR: 0.79 [0.56-1.10], p = 0.160) [5]. Bevacizumab has been also studied in combination with oxaliplatin-based regimens in the NO16966 study, where about 1400 mCRC patients were randomly assigned according to a 2 × 2 design, to receive either FOLFOX or XELOX plus bevacizumab Clomifene or placebo as first-line treatment [6]. The addition of bevacizumab was associated with significantly longer PFS (HR: 0.83 [0.72-0.95], p = 0.0023), that translated into

a trend toward better OS, though not reaching the statistical significance (HR: 0.89 [0.76-1.03], p = 0.077). The magnitude of the effect of bevacizumab seemed less prominent in this experience, when compared with results achieved in the AVF2107 study. The frequent discontinuation of the anti-VEGF together with chemotherapy before disease progression and not for bevacizumab-related toxicity was suggested by authors as a possible explanation for such finding. On the basis of these results, the choice of bevacizumab in the routine upfront approach to the treatment of mCRC is extremely frequent. In fact, it has been demonstrated relatively safe in association with both irinotecan- [7] and oxaliplatin-containing regimens [8] and its specific toxicity profile appears manageable, by applying appropriate clinical selection criteria [9]. Moreover, differently from the anti-EGFR antibodies, the anti-VEGF may be proposed to all patients, without any molecular restriction. However, in spite of its wide use, the magnitude of the benefit derived by the addition of bevacizumab to conventional cytotoxics is still controversial.

At the same time, we advise caution against rendering a certain d

At the same time, we advise caution against rendering a certain diagnosis in the absence of sufficient, confirmatory clinical information. With the data provided in their 2006 report, the clear confidence Harber et al. displayed appears to us to be unwarranted.”
“Introduction In most of the 30 countries Aurora Kinase inhibitor joint in OECD (Organization for Economic Cooperation and Development), the mean age of workers increases as a result of demographic and social trends (Keese et al. 2006).

Birth cohorts since the 1960s are smaller than previous ones, and nowadays a large proportion of the find more youngest age group (15–25 years) in the labour force is still in education. As an additional effect of these trends, the number of available workers will diminish in the next decades. Estimations in the Netherlands for 2025, compared to 2008, show that the number of persons

available for work will decrease by 4.1% (around 340,000 employees) (http://​www.​statline.​nl). Comparable trends are predicted for other Western countries. Participation of a larger part of the people who potentially are able to work is necessary to prevent scarcity on the labour market. The European Council in Lisbon (in 2000) and Stockholm (in 2001) have set ambitious targets to be reached by 2010: to increase the general employment rate MM-102 price to 70% and the employment rate of older workers (55 and older) to 50% (Hutsebaut 2005). Encouraged by these targets and urged by the predicted scarcity in the labour market, many governments have enacted, among others,

measures to discourage Protein kinase N1 early retirement, in order to increase labour force participation (Hutsebaut 2005). In the Netherlands, these measures are rather successful: over the past 15 years, the participation of older workers (aged between 55 and 64) has increased from the all-time low of 24% in 1993 (Wilthagen 2004) to 47% in 2008 (Janssen and Souren 2009). Retirement at a more advanced age will contribute to the trend that a larger number of employees will be of 55 up to 65 years. For a good HRM and occupational health policy it is important to get a better picture of how people in this age group perceive their work and to evaluate what contributes to their job satisfaction, compared to employees in younger age groups. The latter is also important because low job satisfaction is one of the factors that affect the intention to leave (Irvine and Evans 1995; Karatepe 2007; McCarthy 2007) and to early retirement (Sibbald et al. 2003). Moreover, Faragher et. al. (2005) concluded from a meta-analysis that job satisfaction influences the health and well-being of workers. This article addresses employees’ work characteristics, and the relationships between work characteristics and job satisfaction.

pestis travels from the site of infection to draining lymph nodes

pestis travels from the site of infection to draining lymph nodes (LN) prior to disseminating throughout the rest of the body [15, 16]. Bacterial burden data from these experiments give a snapshot of a very narrow window (a specific organ at a specific time) through the course of infection. Furthermore, the approach is invasive, requires a large number of animals, and animals must be sacrificed at each

time point making it impossible to keep track of the progression of infection selleck chemicals llc on the same group of individuals. In vivo bioluminescence imaging (BLI) is an approach that has been used to detect light-emitting cells inside of small mammals [17]. Using BLI, researchers have described and studied dissemination of viral, parasitic and bacterial pathogens within a host in a non-invasive manner [18–21]. Thus, the same group of animals can be imaged for as long as desired over the course of infection. The system requires that the pathogen produce luminescence, and infected animals are then imaged with a high-sensitivity camera that detects very small amounts of light. Non-luminescent bacteria can be genetically modified to express

the lux genes (luxCDABE), which encode a bacterial luciferase and other enzymes that are necessary to generate substrate for luciferase [22]. In the presence of oxygen, luciferase catalyzes a reaction that produces light as a byproduct [23]. We transformed Y. pestis CO92 with plasmid pGEN-luxCDABE that Savolitinib chemical structure contains the luxCDABE genes [24]. Using this strain of Y. pestis expressing the lux genes we determined that it is suitable for in vivo BLI after subcutaneous, intradermal and intranasal inoculation. find more In addition, we determined that BLI is suitable for the study of mutant strains that are attenuated or defective in dissemination or colonization during infection. This extends the findings of a recent report demonstrating

the suitability of BLI to study Y. pestis infections by the subcutaneous route of inoculation [25]. BLI technology offers a new perspective to study the spread of Y. pestis in the host. This technology could be adopted in the future as an alternative to experiments that measured bacterial burdens in specific organs, facilitating the discovery Niclosamide and study of genes that are important in pathogenesis. Results The pGEN-luxCDABE vector is stable in Y. pestis during infection Bacteria carrying a reporter plasmid could potentially lose it at a specific site or time point during infection. A subpopulation lacking the plasmid could result in false negatives or decreases in signal detection that are not necessarily related to lower numbers of bacteria. To determine if pGEN-luxCDABE (pGEN-lux) was maintained during Y. pestis infections, we performed a kinetic study with mice infected with CO92 carrying pGEN-lux. Mice were inoculated subcutaneously (SC) and LN harvested at 24 hours post inoculation (hpi), LN and spleens harvested at 48 and 72 hpi, and LN, spleens and lungs harvested at 96 hpi.

02% (60:40:9; v/v/v), after development, the plates were dried, s

02% (60:40:9; v/v/v), after development, the plates were dried, soaked in 0.5% polymethacrylate in hexane, dried, and blocked for 2 h with 1% of BSA in PBS. Plates were then incubated with mAb MEST-3 PF 2341066 overnight followed by sequential incubations with rabbit anti-mouse IgG and 125I-labeled protein A (2 × 107 cpm/50 ml of BSA/PBS). VRT752271 manufacturer Indirect immunofluorescence Fungi were fixed with 1% formaldehyde in PBS for 10 min. Cells were washed, suspended in 1 ml of PBS, and 20 μl of the solution was added to a coverslip pre-treated with poly-L-lysine 0.1% during 1 h. Air

dried preparations were soaked for 1 h in PBS containing 5% of BSA, and incubated subsequently with culture supernatant of mAb MEST-3 (2 h), biotin-conjugated goat anti-mouse IgG (1 h), and with avidin-conjugated fluorescein (1 h). After each incubation

the coverslips were washed five times with PBS. The coverslips were examined with an epifluorescence microscope [13]. Control experiments for each fungus were carried out, in the presence of an irrelevant monoclonal antibody, and no fluorescence was observed. Cell growth To evaluate the influence of mAbs directed to GSLs on the growth of different fungi, yeasts (104/ml) were incubated in 96-well plate in the presence of mAbs MEST-1, -2, or -3 for 24 h at 37°C, in concentration ranging from 2.5 to 50 μg/ml. The growth rate was evaluated by two procedures; 1) viable CFU were evaluated by plating 100 μl of the samples onto BHI or PGY agar plates, followed by incubation Immune system for 2 days at 37°C, and colony counting; or 2) 5 μl of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium selleck kinase inhibitor bromide (MTT) solution (5 mg/ml MTT in phosphate-buffered saline, pH 7,4) were added to each well and the plates were further incubated at 37°C, for 3 h, after incubation the medium containing MTT was partially removed, and dimethyl sulfoxide (100 μl) was added to solubilize the MTT formazan product [41]. The absorbance of each well was measured at 580 nm by a microtiter ELISA plate reader. Control systems were similarly treated with an irrelevant immunoglobulin

(normal mouse total Ig) and plated. All experiments were repeated three times in triplicates, and the results shown are a representative of these experiments. Fungal differentiation – yeast to mycelium 104 viable yeasts were suspended in 1 ml of PGY (P. brasiliensis) or BHI (H. capsulatum and S. schenckii) medium. The suspension was incubated in a 24-well plate and supplemented with mAb MEST-1, -2, or -3 (at a concentration of 2.5, 10, 25 or 50 μg/ml), after one hour at 37°C, 24-well plate was transferred to a 24°C incubator and kept for 2 days. The number of yeast showing hyphae growth was counted, and presented as percentage of those incubated with irrelevant immunoglobulins (normal mouse total Ig). In control experiment 100% of yeast showed hyphae formation.

RNA was analyzed by semi-quantitative reverse-transcription PCR

RNA was analyzed by semi-quantitative reverse-transcription PCR. PCR products were analyzed on 1.5% agarose ATM/ATR activation gels, stained with ethidium bromide and subsequently visualized. To confirm equal loading, PCR for 16S rRNA was performed in parallel. Ctrl indicates control reactions with no cDNA templates. Because lactoferrin rather than transferrin is the primary carrier of iron on mucosal surfaces and lactoferrin binding proteins are thought to be important virulence factors in some gram-negative bacteria [28], we investigated whether cold shock affects the expression

of these genes. As shown in Figure 2, cold shock increased the mRNA level of lbpB and lbpA genes in strain O35E after 3 h of incubation at 26°C (Figure 2C). Furthermore, cold shock increased the transcriptional level of lbpA and lbpB of other clinical isolates indicating that this effect is a general characteristic of M. catarrhalis (Figure 2D). Enhanced binding of transferrin and lactoferrin on the surface of M. catarrhalis induced by cold shock Because a temperature drop from 37°C to 26°C induces an increase in the copy numbers of genes involved in iron selleckchem acquisition, we investigated whether it also affects the binding

to human transferrin and lactoferrin. Strain O35E and its TbpB-deficient mutant were exposed to 26°C or 37°C and evaluated for their ability to bind transferrin. Binding to transferrin was increased when bacteria were exposed to 26°C (Figure 3A and 3B). The absence of TbpB reduced binding to transferrin, indicating that TbpB is required for maximum binding of transferrin on the surface of cold KU-57788 nmr shock-induced M. catarrhalis. Figure 3 Increase in the binding of transferrin on the surface of M. catarrhalis as a result of cold shock. A, strain O35E and its isogenic mutant O35E.tbpB exposed to 26°C or 37°C for 3 h were incubated with fluorescein isothiocyanate (FITC)-conjugated transferrin

(0.1 μg/mL) and flow cytometry analysis was performed. Shown are representative flow cytometry profiles of strain O35E and O35E.tbpB after exposure selleck at 26°C (gray) or at 37°C (black), which demonstrate that TbpB is required for maximum binding of transferrin on the surface of cold shock-induced Moraxella catarrhalis. The dotted line represents the negative control (bacteria only). The mean fluorescence intensity ± 1 standard deviation for three experiments performed is shown in panel B. *, P< 0.05 for 26°C versus 37°C (one-way analysis of variance). Binding to lactoferrin in a whole-cell solid-phase binding assay was significantly increased when bacteria were exposed to 26°C, in comparison with exposure to 37°C (Figure 4A). The surface binding of human salivary and milk lactoferrin (sLf and Lf, respectively) was further quantitated using flow cytometry, resulting in a clear shift of fluorescence intensity for M. catarrhalis exposed at 26°C (Figure 4B).

The mean and standard errors were determined from 6 qRT-PCR react

The mean and standard errors were determined from 6 qRT-PCR reactions per chromate treatment (3 independent cultures × 2 reactions per culture). Significant differences among chromate treatments for each gene were determined by generating least square means in PROC GLIMMIX with the LS MEANS option in SAS version 9.1. Multiple comparisons were adjusted using Tukey’s test. To normalize the variance of the model residuals, a negative binomial distribution was used for each set of gene expression data. Chromium content in chromate-exposed

cells Arthrobacter strains FB24 and D11 were grown to mid-log phase (OD600, PXD101 ~0.2) in 50 ml 0.2X NB at which time four replicate cultures were amended with 2 mM chromate (final concentration). One culture per strain was incubated without chromate. All cultures were incubated for an additional 2 h. Aliquots of 40 ml of cells were harvested by centrifugation and washed 4 times with ddH2O. Cell pellets were solubilized in concentrated nitric acid (cHNO3) and heated at 95°C for 2 h. Samples were adjusted to a final concentration of 2% HNO3 with double distilled water and analyzed for total chromium content at the Purdue University Mass Spectrometry Center. The 52Cr inductively coupled argon plasma mass spectrometry (ICPMS) results were obtained using an

ELEMENT-2 (ThermoFinnigan, Bremen, Germany) mass spectrometer in the medium resolution mode. The samples were introduced into the plasma using an Aridus desolvating system with a T1H nebulizer (Cetac Technologies, Omaha NE), which is used to enhance sensitivity and reduce oxide and hydride interferences. The argon sweep gas and nitrogen of the Aridus is Torin 2 supplier adjusted for maximum peak height and stability using 7Li, 115In and 238Upeaks obtained from a multi-element standard (1 ng/ml, Merck & Co.). Chromium concentration was normalized per mg protein. Total soluble Methane monooxygenase cell protein concentration was determined using the Lowry method [57] after collecting cells by centrifugation and

extracting protein with 1N NaOH at 100°C. Student’s t-test was used to determine statistically significant differences in the average chromium content between strains D11 and FB24 at the 95% confidence level. Acknowledgements This work was supported by a grant from the Department of Energy’s Environmental Remediation Science Program (grant DE-FG02-98ER62681). K.H. received support from the Purdue Research Foundation and the Purdue Graduate School Bilsland Doctoral Fellowship. We would like to thank Karl Wood and Arlene Rothwell of the Purdue Mass Spectrometry Center for performing the ICP-MS analysis, Jillian Detweiler for assistance with statistical analyses and Gene Wickham, Kurt Jerke for phylogenetic and technical assistance and Militza MEK activity Carrero-Colon for thoughtful discussion. Vector pART2 was a kind gift from Cristinel Sandu. Electronic supplementary material Additional file 1: Supplemental Figure S1.