Secondly, education should also focus on the benefits of TTL activation versus harm of “under-call”. Lastly, ongoing audits should target TTL activation rate

and timely feedback should be provided to all selleck compound players in trauma resuscitations to ensure proper and consistent TTL activation. Attrition of ATLS knowledge may also have contributed to poor compliance. In a study by Ali et al. [6], significant attrition rates of cognitive knowledge and skills was evident as early as 6 months after participants completed an ATLS course. The same group showed the attrition rate was higher for participants from low-volume centers compared to high-volume centers [7]. To address this issue, continued trauma education for all members of the trauma team should be actively encouraged and supported. This can take the form of multidisciplinary trauma simulations, maintenance of ATLS certification, other advanced courses in trauma, and attendance at trauma conferences. Additional training in trauma team crisis resource management may improve team cohesiveness, and the requirement of all physicians involved in trauma resuscitations to maintain active ATLS certification should also be established. This buy Bucladesine study has a number of limitations. Trauma resuscitations are highly dynamic and as such not all actions performed were adequately documented

with certainty. The chart review revealed a lack of time entries in many areas and this has made time-dependent outcome measures hard to gather. In particular, the rate of completion of FAST exams and time to FAST exam could not be reliably obtained from the chart review due to inconsistent record keeping. The study only reviewed data from a one-year period and as a result may not have the necessary power to show differences in major outcomes between the TTL compared to the non-TTL groups. However, we have obtained important data on the performance outcomes in the Casein kinase 1 form of ATLS compliance

rate, readmission rate, and indirect measure of efficiency of trauma resuscitations via times to diagnostic imaging. Additionally, we have also identified areas of future improvement with this quality assessment, and hope that other institutions will use our study as a model to promote their own quality reviews. Conclusions We have demonstrated that TTL involvement significantly improved compliance with many aspects of ATLS, and increased the efficiency of trauma resuscitations by decreasing mean time to diagnostic imaging. There is an acute need to improve compliance with ATLS protocols at our center as well as increase TTL involvement in major traumas at our institution. The reluctance in the hospital culture to activate the trauma team and TTL should be targeted with education around the importance of trauma team activation and involvement of TTL, as well as promotion of a culture of safety.

MFs have the same correlation relation as the resonator mode. Actually, we have neglected the regular fermions (i.e. normal electrons) in the nanowire that interact with the QD in the above discussion. To describe the interaction between the normal electrons and the QD, we use the tight-binding Hamiltonian of the whole wire as [55, 56] , where c k and are the regular fermion annihilation and creation operators with energy ω k and momentum obeying the anti-commutative relation

and ζ is the coupling strength between the normal electrons and QD (here, for simplicity, we have neglected the k-dependence of ζ as in [57]). To go beyond weak coupling, the Heisenberg operator can be rewritten as the sum of its steady-state mean value and a small fluctuation with zero mean value: signaling pathway , , f M =f M0+δ f M and N=N 0 +δ N. Since the driving fields are weak, but classical coherent fields, we will identify all operators with their expectation Trichostatin A concentration values, and drop the quantum and thermal noise terms [31]. Simultaneously, inserting these operators into the Langevin equations (Equations 1 to 4) and neglecting the nonlinear term, we can obtain two equation sets about the steady-state mean value and the small fluctuation. The steady-state equation set consisting of f M0, N 0

and is related to the population inversion ( ) of the exciton which is determined by . For the equation set of small fluctuation, we make the ansatz [54] , 〈δ S -〉=S + e -i δ t +S – e i δ t , 〈δ f M 〉=f M+ e -i δ t +f M- e i δ t , and 〈δ N〉=N + e -i δ t +N – e i δ t . Solving the equation set and working to the lowest order in E pr but to all orders in E pu, we can obtain the nonlinear optical susceptibility as , where and χ (3)(ω pr) is given by (5) where b 1=g/[i(Δ MF-δ)+κ

MF/2], b 2=g/[ i(Δ MF+δ)+κ MF/2], , , , , , d 2=i(Δ pu-δ+ω m η N 0)+Γ 2-g b 1 w 0-d 1 h 2, , d 4=i(Δ pu+δ+ω m η N 0)+Γ 2-g b 2 w 0-d 3 h 5 (where O ∗ indicates the conjugate of O). The quantum Langevin equations of the normal electrons coupled to the QD have the same form as MFs; therefore, we omit its derivation Mirabegron and only give the numerical results in the following. Numerical results and discussions For illustration of the numerical results, we choose the realistic hybrid systems of the coupled QD-NR system [40] and the hybrid semiconductor/superconductor heterostructure [15–17, 20]. For an InAs QD in the coupled QD-NR system, the exciton relaxation rate Γ 1=0.3 GHz, the exciton dephasing rate Γ 2=0.15 GHz. The physical parameters of GaAs nanomechanical resonator are (ω m , m, Q)=(1.2 GHz, 5.3×10-15 g, 3×104), where m and Q are the effective mass and quality factor of the NR, respectively. The decay rate of the NR is γ m = ω m /Q=4×10-5 GHz.

Nearly identical nucleotide sequences

of nifNE markers were found in different pSym plasmids of the studied population (Figure 6C), confirming the core character of symbiotic genes and their high conservation, despite the overall genome differentiation [11]. The extent of gene adaptation to a given compartment in the host genome was assessed by analyses of alternative codon usage. Three groups of well separated genes were obtained corresponding to the chromosome, chromid-like and ‘other plasmids’ genome compartments (Figure 7A) with 96% accordance with hybridization data. In conclusion, the sequence divergence of particular genes may be affected by their location in the given genome compartment. When all the sequences of the individual strains studied were subjected to a discrimination H 89 price analysis, we obtained good separation of K3.22 and a group of strains related to RtTA1 (Figure 7B) that formed the outermost branch in the phylogenic tree. The remaining strains were randomly mixed with each other but apparently separated from K3.22 and TA1-related strains, which suggested Doramapimod price no differences in codon usage within the main group. The CAI analyses of the evaluated

sequences confirmed good adaptation of chromosomal and chromid-like genes (high CAI values) to host genomes and lower CAI values for ‘other plasmids’ genes. The CAI values also reflect the level of transcriptional and translational activity of particular genes [29]. While the activity of most of the chromosomal and chromid-like genes could be considered at least to some extent constitutive, the ‘other plasmids’ and especially symbiosis-related genes are expressed only transiently in the symbiotic stage [42]. Therefore, in the Rhizobium model, the differences in codon usage in translation reflect the balance between the selection pressure and random mutations in the functionally differentiated genome compartments. The differences in codon usage and CAI values between the genome compartments are most likely a consequence of differential gene expression and adaptability to optimal codon usage in host genomes [42]. Conclusion Our study showed

that, even within a small rhizobial however population of clover nodule isolates, substantial divergence of genome organization can be detected especially taking into account the content of extrachromosomal DNA. Despite the high variability with regard to the number and size of plasmids among the studied strains, conservation of the location as well as the dynamic distribution of the individual genes (especially replication genes) of a particular genome compartment was demonstrated. The sequence divergence of particular genes may be affected by their location in the given genome compartment. The ‘other plasmid’ genes are less adapted to the host genome than the chromosome and chromid-like genes. Acknowledgements and Funding This work was supported by Grant No. N N301 028734 from Ministry of Science and Higher Education of Poland.

In each case, the reaction was allowed to proceed at 37°C for 0, 7.5, 15, 22.5, and 30 min as described in a previous section. Statistical analysis Statistical and curve-fitting analyses were performed using Prism 4.0 (GraphPad Software Inc.). The data are expressed as means ± SEM. Differences between groups were assessed by one-way analysis of variance (ANOVA), followed by Student–Newman–Keul’s test. Values of percentage inhibition of EM degradation were calculated using following formula, which was described earlier (Tomboly et al., 2002): $$ \textInhibition \left( \% \right)

\, = \, \left( k_0 – k_\texti \right)/k_0 \times 100, $$where k 0 the rate constant of degradation without inhibitor, k i the rate constant of degradation with inhibitor. Results Effect of inhibitors on degradation of EMs by DPP IV We evaluated EMDB-2 and EMDB-3 for their inhibitory effect on degradation TGF-beta family of EMs by DPP IV. Diprotin A was included in the study for comparison. Degradation of EMs was analyzed by reversed phase HPLC. Effects of 30 min incubation of EM-2

with DPP IV in the absence and presence of inhibitors are shown in Fig. 2. The chromatographic peak area of EM-2 was found to decrease greatly in the sample without inhibitors. Diprotin A almost completely suppressed enzymatic cleavage of EM-2, while EMDB-2 and EMDB-3 only partially protected EM-2 against hydrolysis. Degradation rates and half-lives of EMs alone and in the presence of inhibitors are collected in Table 1. Different rates of degradation of EM-1 and EM-2 by DPP IV were observed. EM-1 was Selleckchem Captisol about 1.5 times more resistant to DPP IV than EM-2, which is in agreement with the data obtained by others (Tomboly et al., 2002; Grass et al., 2002; Fujita and Kumamoto, 2006; Keresztes et al., 2010). EMDB-2 and EMDB-3 increased EM-1 and EM-2 half-lives two- to threefold. The effects of inhibitors on degradation of EMs after 30 min incubation with DPP IV are summarized in Table 2. EMDB-3 appeared to be a better Sodium butyrate DPP IV inhibitor than EMDB-2. The Lineweaver–Burk plots revealed that both tested compounds acted as competitive inhibitors of DPP IV (Fig. 3). Fig. 2 Effect

of inhibitors on the degradation of EM-2 by DPP IV. The reaction mixture was incubated at 37°C for 30 min in the absence (a) and presence of diprotin A (b), EMDB-2 (c), and EMDB-3 (d). Asterisk indicates the peak derived from the inhibitor added Table 1 Degradation rates (k) and half-lives (t 1/2) of EMs incubated with DPP IV alone and in the presence of inhibitors Inhibitor DPP IV EM-1 EM-2 100 × k (1/min) t 1/2 (min) 100 × k (1/min) t 1/2 (min) Without inhibitor 4.12 ± 0.2 16.7 ± 0.52 6.30 ± 0.31 10.9 ± 0.64 Diprotin A 0.13 ± 0.01 530 ± 14.5*** 0.18 ± 0.01 383 ± 20.2*** Tyr-Pro-Ala-NH2 (EMDB-2) 3.02 ± 0.09 22.9 ± 1.14* 3.48 ± 0.13 19.8 ± 0.75* Tyr-Pro-Ala-OH (EMDB-3) 2.51 ± 0.12 27.5 ± 1.21* 2.52 ± 0.13 27.4 ± 1.41* * P < 0.05, *** P < 0.

Several reports indicated that H. pylori has the ability to form biofilms on abiotic surfaces in vitro as well as on human gastric mucosa [18–21, 23]. The results of the biofilm formation analyses demonstrated that strain TK1402 has strong biofilm forming ability compared to other strains independent of its growth rate. Development of strain TK1402 and SS1 biofilms from day 1 to day 6 demonstrated that it took 3 days for biofilm maturation under these conditions, suggesting that H. pylori biofilm formation might proceed in an organized fashion

through early (Day 1), intermediate (Day 2) and maturation (after Day 3) phases of development. Similar distinct developmental phases have been reported for biofilm formation by other bacterial species [24, 25]. Since development of biofilms is closely associated with the generation of a matrix, the majority of which is

extracellular material, biofilm development BI 2536 manufacturer in H. pylori appears to share common basic steps with other biofilm forming bacteria. The biofilm forming cells at day 3 generally appeared to be viable when the cells were exposed to Live/Dead BacLight staining. In addition, the normalized CFU values for the biofilm and broth culture cells following 2 days of incubation were comparable. In 3-day biofilm cells, this value was slightly decreased compared to 3-day broth culture cells, suggesting the presence of some dead cells in the biofilm. These results are consistent with the maturation phase of the development of biofilms in 3-day biofilms of strain TK1402, since biofilms see more are thought to be encased in an EPS matrix as well as dead cells [26]. In addition, strain TK1402

exhibited thick biofilm formation. The biofilm DNA ligase morphology of strain TK1402 showed direct cell-cell bound aggregates as well as flagella-dependent binding forms. The cell-cell interacting forms might act as precursors for thick biofilm formation. Gots et al. indicated that cell-cell aggregation induces a multilayered architecture during Staphylococcus epidermidis biofilm formation [27]. Moreover, in our SEM observations, for the majority of the H. pylori strains examined, ie., SS1, biofilms may contain autolysed cells. On the other hand, there were clearly intact cells in TK1402, as well as TK1049, biofilms and the later is also another strong biofilm forming strain. These observations suggested that these strong biofilm forming strains may remain in an active metabolic state for a relatively long time without exhibiting morphological changes or autolysis, in comparison with the other strains. These later properties could be responsible for the weaker biofilm forming activities of most of the strains examined in this study. In the SEM observations of TK1402 biofilms, there were many OMV. OMV production is a physiologically normal function of gram-negative bacteria [22, 28]. It was also reported that the H. pylori strains released OMV into the extracellular space [29, 30].

Infect Immun 1991, 59:1739–1746.PubMed 21. Hijnen M, van Gageldonk PG, Berbers GA, van Woerkom T, Mooi FR: The Bordetella pertussis virulence factor P.69 pertactin retains its immunological check details properties after overproduction in Escherichia coli. Protein Expr Purif 2005, 41:106–112.CrossRefPubMed 22. Lee SF, Halperin SA, Knight JB, Tait A: Purification and

immunogeniCity of a recombinant Bordetella pertussis S1S3FHA fusion protein expressed by Streptococcus gordonii. Appl Environ Microbiol 2002, 68:4253–4258.CrossRefPubMed 23. Roberts M, Fairweather NF, Leininger E, Pickard D, Hewlett EL, Robinson A, Hayward C, Dougan G, Charles IG: Construction and characterization of Bordetella pertussis mutants lacking the vir-regulated P.69 outer membrane protein. Mol Microbiol 1991, 5:1393–1404.CrossRefPubMed 24. Mattoo S, Cherry JD: Molecular

pathogenesis, epidemiology, and clinical manifestations of respiratory infections due to Bordetella pertussis and other Bordetella subspecies. Clin Microbiol Rev 2005, 18:326–382.CrossRefPubMed 25. Hellwig SM, Rodriguez ME, Berbers GA, Winkel JG, Mooi FR: Crucial role of antibodies to pertactin in Bordetella pertussis immunity. J Infect Dis 2003, 188:738–742.CrossRefPubMed 26. Cherry JD, Gornbein J, Heininger U, Stehr K: A search for serologic correlates of immunity to Bordetella pertussis cough illnesses. Vaccine 1998, 16:1901–1906.CrossRefPubMed 27. Storsaeter J, Hallander HO, Gustafsson L, Olin P: Levels of anti-pertussis antibodies CA4P related to protection after household exposure to Bordetella pertussis. Vaccine 1998, 16:1907–1916.CrossRefPubMed 28. Ausiello

CM, Lande R, Stefanelli P, Fazio C, Fedele G, Palazzo R, Urbani F, Mastrantonio P: T-cell immune response assessment as a complement to serology and intranasal protection 17-DMAG (Alvespimycin) HCl assays in determining the protective immunity induced by acellular pertussis vaccines in mice. Clin Diagn Lab Immunol 2003, 10:637–642.PubMed 29. Mills KH, Barnard A, Watkins J, Redhead K: Cell mediated immunity to Bordetella pertussis : role of Th1 cells in bacterial clearance in a murine respiratory infection model. Infect Immun 1993, 61:399–410.PubMed 30. Cheung GY, Xing D, Prior S, Corbel MJ, Parton R, Coote JG: Effect of different forms of adenylate cyclase toxin of Bordetella pertussis on protection afforded by an acellular pertussis vaccine in a murine model. Infect Immun 2006, 74:6797–6805.CrossRefPubMed 31. Medical Research Council: Vaccination against whooping cough: relation between protection in children and results of laboratory tests. Br Med J 1956, 2:454–462.CrossRef 32. Guiso N, Capiau C, Carletti G, Poolman J, Hauser P: Intranasal murine model of Bordetella pertussis infection .I. Prediction of protection in human infants by acellular vaccines. Vaccine 1999, 17:2366–2376.CrossRefPubMed 33.

In all of the loci, the differences in the number of repeats were weighted equally

because at one locus, multiple tandem repeats can be incorporated during one recombination event. The publicly available MLVA database for Brucella (MLVA-NET for Brucella, http://​mlva.​u-psud.​fr/​brucella/​) was used to identify or confirm the identity of all of the isolates used in this study. The comparison between the caliper data and MLVA bank showed some discrepancies for the allelic sequences that were obtained using different electrophoretic techniques. Due to the different nature of the gel matrix, these differences were resolved by sequencing [18, 30]. Culture conditions and sample preparation for MALDI-TOF-MS analysis From a frozen stock, the bacteria were cultured on blood agar plates for at least 48 h at 35°C in the presence of 5% CO2. Entospletinib Before sample preparation, the isolates were re-grown for 48 h at 35°C in the presence of 5% CO2. Sample preparation was performed according to the company guidelines (Bruker Daltonics,

Bremen, Germany). Briefly, 30 colonies were suspended in 300 μl of water (MilliQ, Millipore, Billerica, MA, U.S.) and mixed carefully. Next, 900 μl of absolute ethanol (Fisher Scientific, Loughborough, UK) was added and the suspension was mixed. Subsequently, the suspension was incubated for 90 min to inactivate all of the bacteria. After this inactivation step, the suspension samples were centrifuged Rho for 10 min at 10, 000 g. The supernatant was removed. To remove the www.selleckchem.com/products/Adriamycin.html remaining ethanol residue, the spinning step was repeated, and the remaining supernatant was removed. Subsequently, 50 μl of 70% formic acid was added to the pellet, and the pellet was mixed. Next, 50 μl of pure acetonitrile (LC-MS grade, Fluka/Aldrich, Stenheim,

Germany) was added, and the suspension was mixed carefully. The particulate matter that could not be dissolved was spun down by centrifugation for 2 min at 10, 000 g. Finally, four spots were created, using 0.5 μl of the supernatant per spot, onto a MALDI-TOF target plate (MTP 384 target polished steel #209519, Bruker Daltonics) and air dried. Subsequently, the spots were overlaid with 0.5 μl of α-cyano-4-hydroxycinnamic acid (HCCA, Bruker Daltonics) and a 10 mg/ml acetonitrile/water solution (1:1) with 2.5% trifluoroacetic acid (TFA) (Fluka/Aldrich, Stenheim, Germany) and dried at room temperature. Mass spectra acquisition All of the mass spectra were automatically acquired on a Bruker Autoflex III smartbeam instrument (Bruker Daltonics GmbH, Bremen, Germany) in linear mode using the following parameters: 40% laser intensity, positive polarity, 350 ns PIE delay, 20 kV source voltage 1, 18.7 kV source voltage 2, 8 kV lens voltage, 1.522 kV linear detector voltage, and 800 Da detector gating.

Our finding

that LasRI can also repress Pel expression in strain ZK at 37°C, a temperature relevant to infection, raises the intriguing possibility that QS may trigger dissolution of clinical biofilms. This would be analogous to other bacterial pathogens like Vibrio cholerae [62] and Staphylococcus aureus [63]. Results with the particular strain chosen, ZK2870, are significant, because the autoaggregative behavior of this strain under some environmental conditions appears most representative among clinical and environmental isolates of P. aeruginosa [12]. The observed differences in the colony phenotype of different Pseudomonas strains (Figure 2) might be attributed to the presence or absence of a particular EPS locus or regulatory variability in strains with identical EPS loci. Our second major finding selleck inhibitor is that las QS mediates colony morphology via AQ signaling. Phenotypic analysis along with AQ signal quantitation by TLC suggested www.selleckchem.com/products/pnd-1186-vs-4718.html that a Series A congener is involved. PqsA-D produce at least 8 different compounds within this series [64]. Of these, HHQ and HNQ have been shown to accumulate in a PAO1 lasR mutant [20]. Other prominent AQs, 2,4-dihydroxyquinoline

(DHQ) and 2-heptyl-4-hydroxyquinoline N-oxide (HQNO), that require some of the enzymes encoded by pqsA-D, but are not PqsH substrates, show reduced levels in a lasR mutant compared with the wild-type [20]. Our chemical supplementation experiments indicate that neither HHQ nor HNQ modulate wrinkling. This implies that one of the other less-well characterized Series A congeners have

a role in this process, further expanding the various cellular functions of AQs in P. aeruginosa. A detailed investigation utilizing liquid chromatography/mass spectrometry along with chemical synthesis would be able to identify the compound in question. PqsE, a putative regulator encoded by the pqsA-E operon whose precise function mafosfamide is not known, is unlikely to have a role in AQ-mediated colony wrinkling, because pqsA-D expression in a lasR pqsR mutant that does not express pqsE was sufficient to induce wrinkling (Figure 7B). Interestingly, in Burkholderia pseudomallei the lack rather than the overproduction of the Series A congener HHQ results in a wrinkly colony phenotype [61]. In addition, AQ signal overproduction has previously been shown to induce autolysis in P. aeruginosa populations, forming plaques that result in characteristic translucent zones in colonies [36], different from those we observed. Autolysis appears to be mediated by PQS rather than a Series A congener [65]. Taken together, our data can be rationalized as follows: In the wild-type, both Series A and Series B congeners are produced as LasR activates pqsR and pqsH. In the lasR mutant, Series A congeners accumulate and the Series A to Series B ratio increases because of (1) reduced pqsH expression and (2) presumably lasR-independent expression of pqsR [25] resulting in continued activation of pqsA-E.

6 Top Two-dimensional

thin layer separation of six Plastoquinone C subunits, from tomato, in diisopropyl ether-benzene (15:85) in both directions. Bottom Cochromatography of tomato PQC with spinach PQC 2 and 3 in the same solvent. PQC2 and PQC3 are the major PQCs in spinach and they move with tomato PQC 2 and 3. (After Barr et al. 1967a, b) Extensive study of the distribution of the 12 new isoprene analogs with modified side chains (Fig. 7) was done to see if any of them were available in amounts sufficient to play any role in photosynthesis. Lichtenthaler and Calvin (1964) found PQA in what was called “quantosomes” [this term has now been abandoned—Editor] in the same ratio to chlorophyll as in whole chloroplasts which indicated they were available in the photosynthetic unit. In PSI-7977 manufacturer a personal communication, Calvin informed me that they found no Belnacasan concentration coenzyme Q in chloroplasts. In 17 species, Rita Barr and I (see Barr and Crane 1967) found that PQA and PQC1–C4 were regularly present in significant amounts (over 0.004 M PQ/mg chlorophyll), whereas PQB and PQC5–PQC6 were often missing. The same pattern was

found by Sun et al. (1968) in 21 species, ranging from cyanobacteria to red algae: PQA and PQC1–PQC4 were always present (except in a white strain of Euglena). Several studies have shown that PQA and PQC1–PQC5 increase as plants age (Lichtenthaler 1969). Likewise, an increase of PQA and PQC1–PQC4 occurs during greening of etiolated plants (Barr and Crane 1970). PQB did not appear even after 72 h of light and only in maize did PQC5–PQC6 appear with short exposure to light (Barr and Crane 1970). The quinones that appear in the light are the most likely to be involved in photosynthesis; these

include PQA, PQC1–PQC4, Vitamin K1, and α-TQ. In a few plants, e.g., alfalfa, PQC is missing in winter (Bucke and Hallaway 1966). As pointed out by Amesz (1973), this precludes PQC from the main pathway of photosynthesis but does not eliminate it from its function in side reactions. The assay of PQC and α-TQ is difficult because of incomplete extraction even with acetone which in contrast to PQA indicates tight bonding to some protein (Henninger and Crane 1963). either Another problem with assay for PQC is that 10–30% may be in the reduced form (Kruk and Strzalka 1998). Fig. 7 Structure of plastoquinone A (top), plastoquinone C1 (middle), and plastoquinone B1 (bottom). Epoxidation of the double bond in the second prenyl group from the ring produces a hydroxyl group on the side chain to make PQC1. Successive oxidation of other prenyl groups makes PQC 2, 3, 4, 5, and 6. The PQB1, 2, 3, 4, 5, and 6 groups are produced by esterification of a fatty acid to the hydroxyl groups of the PQCs.

31 ± 0.02a 4.5 ± 0.3 24 ± 1a 33 ± 1a 1.17 ± 0.32a ND 56 ± 4a 35 ± 3a 0.51 ± 0.03a 12 ± 0.4a 22 ± 0.6a 37 ± 2a 2.12 ± 0.43b ND   Stress 45 ± 4a AZD5582 concentration 22 ± 3a 0.34 ± 0.02a 5.0 ± 0.2 22 ± 2a 32 ± 1a 1.24 ± 0.20a ND 31 ± 3b 16 ± 1b 0.34 ± 0.03b 5.3 ± 0.1b 14 ± 0.7b 24 ± 1b 2.37 ± 0.39b ND otsAch Control 48 ± 5a 24 ± 3a 0.37 ± 0.03a

5.5 ± 0.4 27 ± 3a 35 ± 3a 1.15 ± 0.29a ND 61 ± 4a 42 ± 5a 0.52 ± 0.03a 12.5 ± 0.5a 27 ± 1.2a 41 ± 4a 1.90 ± 0.32b ND   Stress 46 ± 5a 25 ± 5a 0.35 ± 0.05a 5.3 ± 0.3 24 ± 1a 35 ± 3a 1.25 ± 0.30a ND 35 ± 5b 19 ± 3b 0.37 ± 0.03b 5.5 ± 0.3b 16 ± 1.5b 25 ± 1b 2.08 ± 0.37b ND Nodules number (NN), nodule dry weight [NDW, (mg plant-1)], plant dry weight [PDW, (g plant-1)], total nitrogen content [TN, (mg plant-1)], acetylene reduction activity [ARA, (μmol C2H4 h-1 g-1 NDW)],leghaemoglobin [Lb, (mg Lb g-1 NDW)], and trehalose (Tre) in bacteroids (B) and

nodule cytosol (C) [μmol gDW-1] content in nodules and plants subjected or not (control) to moderate or severe drought conditions. Values in a column followed by the same lower-case letter are not significantly different as determined see more by the Tukey HSD test at P ≤ 0.05 (n = 9). ND. Not detected. As shown in Table 2, NN and NDW per plant was negatively affected by a severe drought since a decrease of about 45% and 53% in those parameters was observed in plants inoculated with the wild-type strain compared to control plants. A similar decrease of NN (43%) and NDW (49%) was observed in

plants subjected to a severe stress and inoculated with the otsAch mutant compared to control plants (Table 2). After a severe drought, a 53% and 49% reduction of PDW was observed in mafosfamide plants inoculated with the wild-type or the otsAch mutant, respectively. Plants inoculated with any of the strains and subjected to severe drought showed a similar reduction of about 30% in TN compared to control plants (Table 2). Plants inoculated with the wild-type strain and subjected to severe drought showed an inhibition of ARA of about 36% compared to control plants. This activity was similarly dropped in nodules produced by the otsAch mutant under severe drought (41% compared to control plants) (Table 2). A severe drought provoked a significant decline in Lb content of about 35% in plants inoculated with the wild-type strain compared to control plants Likewise, this parameter was also reduced of about 39% in plants inoculated with the otsAch mutant and subjected to a severe drought (Table 2). Finally, trehalose content in bacteroids of the wild type and otsAch strains was similar, regardless of the treatment, suggesting that under symbiotic conditions (i.e. with other trehalose precursors available) other trehalose synthesis genes (i.e. TreS or TreYZ) may be operating. Trehalose was not detected in the cytosol of nodules induced by either the wild type or the otsAch strain under any condition tested, suggesting that trehalose in the R.