Circulation 116:e418–e499CrossRefPubMed

12. Lawrence VA, Hilsenbeck SG, Noveck H, Poses RM, Carson JL (2002) Medical complications STA-9090 solubility dmso and outcomes after hip fracture repair. Arch Intern Med 162:2053–2057CrossRefPubMed 13. Carbone L, Buzkova P, Fink HA, Lee JS, Chen Z, Ahmed A, Parashar S, Robbins JR (2010) Hip fractures and heart failure: findings from the Cardiovascular Health Study. Eur Heart J 31:77–84CrossRefPubMed 14. Lee TH, Marcantonio ER, Mangione CM, Thomas EJ, Polanczyk CA, Cook EF, Sugarbaker DJ, Donaldson MC, Poss R, Ho KK, Ludwig LE, Pedan A, Goldman L (1999) Derivation and prospective validation of a simple index for prediction of cardiac risk of major noncardiac surgery. Circulation 100:1043–1049PubMed 15. Detsky AS, Abrams HB, Forbath N, Scott JG, Hilliard JR (1986) Cardiac assessment for patients undergoing noncardiac surgery. A multifactorial clinical risk index. Arch Intern Med 146:2131–2134CrossRefPubMed 16. Goldman L, Caldera DL, Nussbaum SR, Southwick FS, Krogstad D, Murray B, Burke DS, O’Malley TA, Goroll AH, Caplan CH, Nolan J, Carabello B, Slater EE (1977) Multifactorial index of cardiac risk in noncardiac surgical procedures. N Engl J Med 297:845–850CrossRefPubMed 17. Chambers J (2005) Aortic stenosis. Bmj 330:801–802CrossRefPubMed 18. Lindroos M, Kupari M, Heikkila J, Tilvis R (1993) Prevalence of aortic valve abnormalities

in the elderly: an echocardiographic click here study of a random population sample. J Am Coll Cardiol 21:1220–1225CrossRefPubMed 19. Stewart BF, Siscovick D, Lind BK, Gardin JM, Gottdiener JS, Smith VE, Kitzman DW, Otto CM (1997) Clinical factors associated with calcific aortic valve disease. Cardiovascular health study. J Am Coll Cardiol 29:630–634CrossRefPubMed 20. McBrien ME, Heyburn G, Stevenson M, McDonald S, Johnston NJ, Elliott JR, Beringer TR (2009) Previously undiagnosed aortic stenosis revealed by auscultation in the hip fracture population—echocardiographic findings, management and outcome. Anaesthesia 64:863–870CrossRefPubMed 21. Network SIG (2009) Management

of hip fracture in older people. pp 1–48 22. Adunsky A, Kaplan A, Arad M, Mizrahi EH, Gottlieb S (2008) Aortic stenosis in elderly hip isothipendyl fractured patients. Arch Gerontol Geriatr 46:401–408CrossRefPubMed 23. Bartels C, Bechtel JF, Hossmann V, Horsch S (1997) Cardiac risk stratification for high-risk vascular surgery. Circulation 95:2473–2475PubMed 24. Myers J, Do D, Herbert W, Ribisl P, Froelicher VF (1994) A nomogram to predict exercise capacity from a specific activity questionnaire and clinical data. Am J Cardiol 73:591–596CrossRefPubMed 25. Nelson CL, Herndon JE, Mark DB, Pryor DB, Califf RM, Hlatky MA (1991) Relation of clinical and angiographic factors to functional capacity as measured by the Duke activity status index. Am J Cardiol 68:973–975CrossRefPubMed 26.

Bacterial loads in cecum content (A), mLN (B), spleen (C) and liver (D)

were assessed by plating at day 4 p.i.. n.s., statistically not significant; *, statistically significant (p < 0.05, Two-way ANOVA). MT4 protects wild-type C57BL/6 mice when challenged with wild-type S. Typhimurium The immunogenic potential of MT4 in wild-type C57BL/6 mice was analyzed by previously established BAY 57-1293 molecular weight vaccination and challenge protocol using TTSS-2 deficient S. Typhimurium strain [34]. Three groups of wild-type C57BL/6 mice were vaccinated with MT4 (n = 10), MT5 (n = 10) and PBS (negative control; n = 10). The fecal shedding was analyzed as a measure of cecal colonization during vaccination period. Both, MT5 and MT4 strains reached a bacterial load of ~109 CFU/g (of cecal content) in the gut lumen at the day 1 p.v.; however, the bacterial loads slightly declined at day 14 and day 28 p.v. (Figure 2A). Half the number of vaccinated mice (MT5, n = 5; MT4, n = 5; PBS, n = 5) were sacrificed Doxorubicin order to analyze cecal inflammation and the colonization levels in different systemic sites at day 30 p.i. With both the strains, cecum colonization was maintained up to ~107-9 CFU/g. The bacterial load

in mLN was lower as compared to the acute infection experiments (compare Figure 1B to 2B) whereas cecal mucosa did not show any sign of disease (Figure 2C). The remaining mice were analyzed for protection against a challenge with wild-type S. Typhimurium. At day 30 p.v., the remaining vaccinated mice (MT4, n = 5; MT5, n = 5; PBS, n = 5) were treated with 20 mg of ampicillin to remove regrown gut flora and any residual vaccine strain. Mice groups were then challenged with wild-type S. Typhimurium at day 31st (200 CFU by gavage). The wild-type S. Typhimurium was able to colonize the lumen efficiently and reached the carrying capacity by day 3 p.c. in all three immunized groups (Figure 3A). Mice in the PBS treated control group suffered from severe enteropathy (Figure 3B). In contrast, Reverse transcriptase the mice immunized with MT5 and MT4 strains did not show any signs of mucosal inflammation (Figure 3B). Furthermore, spleen and liver colonization by wild-type S. Typhimurium was significantly

reduced in both the vaccinated groups (p < 0.05; Figure 3A). Thus, the data indicates that MT4 strain conferred equivalent level of protection from Salmonella inflicted disease as MT5 strain. Figure 2 Vaccination experiment analyzing the attenuation of MT4 at day 30 p.v. For vaccination, C57BL/6 mice were treated with PBS (n = 10; grey solid circles), MT5 (5x107CFU; n = 10; black solid circle) and MT4 (5×107 CFU; n = 10; open circle). (A) Fecal shedding as analyzed by plating. PBS-controls: below detection limit (stripped line); (B) Colonization by the vaccine strains (MT5, n = 5 and MT4, n = 5) in cecal content, mLN, spleen and liver; (C) Cecal pathology at day 30 p.v.. n.s., not significant; *, statistically significant (p < 0.05).

Methods Participants Fourteen healthy untrained males (22.1 ± 2.3 yrs, 173 ± 7.7 cm, 76.2 ± 9.3 kg) volunteered for this study. Descriptive characteristics of the participants are presented in table 1. To meet the criteria the men (a) were non-smokers; (b) had not participated in resistance-training, or any form of structured exercise, for at least six months; (c) had not ingested any ergogenic supplement for a 24-week period

prior to the start of supplementation; and (d) agreed not to ingest any other nutritional supplements, or non-prescription drugs that may affect muscle re-growth during the study. In addition, participants agreed to refrain from using any remedy (i.e. massage, ultrasound etc.) for muscle soreness other than

consumption of the supplement VX-765 cost given; and agreed not to participate in any form of physical activity 2 weeks prior to supplementation and during the 2 week recovery period. All participants were informed verbally, as well as in writing, as to the objectives of the experiments, together with the potential associated risks. All participants signed an informed consent document approved by the Human Research Ethics Committee of Victoria University of Australia. All procedures conformed to National Health and Medical Research Council guidelines for the ethical conduct of research involving humans. Table 1 Participant baseline LY2157299 price characteristics Characteristics CHO Cr-CHO P-value Age (yrs) 21.7 ± 3 22.6 ± 2 0.52 Weight (kg) 74.4 ± 7 77.9 ± 12 0.51 Leg Press 1 RM (kg) 85.9 ± 16 83.62 ± 15 0.80 Leg Extension 1 RM (kg) 40 ± 10 36.4 ± 10 0.49 Leg Flexion 1 RM (kg) Extension 26.8 ± 16 34.1 ± 13 0.35 Data are means ± standard deviations of mean. SI unit conversion factor: 1 kg = 2.2 lbs Experimental design All procedures were completed at the Human Performance Laboratory at Victoria University. Two weeks prior to baseline testing, participants underwent

1 repetition maximum (RM) strength assessments on the dominant limb and a familiarisation session of the equipment that would be utilized to assess muscle performance. The dominant limb would undergo the damage Selleck Lenvatinib protocol, while the contralateral limb served as the control. Participants were randomised in a double-blind placebo-controlled fashion into 2 groups: carbohydrate-only (CHO) (n = 7) or Cr-carbohydrate (Cr-CHO) (n = 7), and issued with their supplement and dosing instructions. On day 1, participants arrived at the laboratory in the morning and underwent baseline performance assessments and blood sampling. Participant’s then underwent catherization of the forearm vein and performed an exercise session designed to cause damage to the knee extensor and flexor muscles. Blood samples were taken at 30 minutes, 1, 2, and 4 hours following the bout of exercise.

e. O. fusispora (Seaver) E. Müll., S. pachythele, X. leve, and X. verrucosum. Huhndorf (1993) formally transferred S. applanata Petch and S. pachythele to Xenolophium. Phylogenetic study Phylogenetic analysis based on LSU sequences indicated that Ostropella albocincta clusters together with Xenolophium applanatum as well as species of Platystomum, but they receive poor support (Mugambi and Huhndorf 2009b). They all were temporarily assigned under Platystomaceae (Mugambi and Huhndorf 2009b). Concluding remarks Although the placement of Ostropella albocincta under Platystomaceae lacks support, Ostropella should be excluded from

Melanommataceae despite its trabeculate pseudoparaphyses. Paraliomyces Kohlm., Nova Hedwigia 1: 81 (1959). (Pleosporales, genera incertae sedis) Generic description Habitat marine, saprobic. Ascostromata immersed, penetrating into the substrate GDC-0980 price with dark brown hyphae. Ascomata medium-sized, solitary, immersed or erumpent, www.selleckchem.com/products/GDC-0449.html subglobose to pyriform, subiculate or nonsubiculate, papillate or epapillate, ostiolate, periphysate, carbonaceous. Peridium thick. Hamathecium of long trabeculate pseudoparaphyses. Asci 8-spored, bitunicate, fissitunicate, cylindrical, with a short furcate pedicel, without apical apparatus, uniseriate. Ascospores ellipsoid to broadly fusoid with broadly rounded ends, 1-septate, constricted at the septum, hyaline, smooth-walled, surrounded by a gelatinous sheath. Anamorphs reported for genus: none.

Literature: Kohlmeyer 1959; Tam et al. 2003. Type species Paraliomyces lentifer Kohlm. [as ‘lentiferus’], Nova Hedwigia 1:

81 (1959). (Fig. 73) Fig. 73 Paraliomyces lentifer (from Herb. J. Kohlmeyer No. 1720). a Section of an immersed ascoma. b Eight-spored cylindrical asci embedded in pseudoparaphyses. c, d Cylindrical Cobimetinib concentration asci with short pedicels. e–h One-septate hyaline ascospores. Scale bars: a = 100 μm, b–d = 20 μm, e–h = 10 μm Ascostromata black, immersed, penetrating into the substrate with dark brown hyphae. Ascomata up to 680 μm high × 540 μm diam., solitary, immersed or erumpent, subglobose to pyriform, subiculate or nonsubiculate, papillate or epapillate, ostiolate, periphysate, carbonaceous (Fig. 73a). Peridium thick. Hamathecium of long trabeculate pseudoparaphyses, 1–1.5 μm broad. Asci 90–130 × 12–17 μm (\( \barx = 116 \times 15\mu m \), n = 10), bitunicate, fissitunicate, cylindrical, 8-spored, uniseriate, with a short furcate pedicel, without apical apparatus (Fig. 73b, c and d). Ascospores 17.5–25 × 10–12.5 μm (\( \barx = 21 \times 11\mu m \), n = 10), ellipsoid to broadly fusoid with broadly rounded ends, 1-septate, constricted at the septum, hyaline, smooth-walled, surrounded by a gelatinous sheath that contracts to form a lateral, lentiform, viscous appendage over the septum, 7.5–12.5 μm diam., 1–3 μm thick (Fig. 73e, f, g and h). Anamorph: none reported. Material examined: USA, Florida, Charlotte Harbor in Punta Garda, 10 Jan. 1964, leg., det. J.

Figure 3 presents the scatter plots of the event residence time t d versus blockade current amplitude ∆I for different experiment conditions. Once a DNA strand enters the nanopore, it will block the ions in and out the nanopore and cause ionic current reduction. The amplitude of the blocked ionic current can be expressed with Kowalczyk’s

model [31], (2) where V is the applied bias voltage, and is the effective diameter of the nanopore with DNA in the pore. According to formula (2), the blocked ionic current amplitude (∆I) is linearly proportional to σ for the nanopore with the same diameter. Therefore, the amplitude of the blockade ionic current for DNA translocation through the nanopore in MgCl2 solution is expected to

be larger than that in the KCl buy BGB324 solution with the same molar concentration because the former has a high electrolytic conductivity. Unfortunately, Trichostatin A purchase the results as shown in Figure 3 do not meet such prediction. The 20-nm diameter nanopore produced a little difference in the amplitude of the blocked ionic current in the three salt solutions (1 M KCl, 0.5 M KCl + 0.5 M MgCl2, and 1 M MgCl2). As shown in Figure 3, the red solid circle points denote the events for the 48.5 kbp λ-DNA translocation through the nanopore in 1 M KCl solution. The green solid triangle points stand for the events that occurred in 0.5 M KCl + 0.5 M MgCl2 solution, and the black open rectangle symbols stand for the events in 1 M MgCl2 solution. The three symbols almost overlap with the black open rectangle symbols which are located a little higher. This result tells us that the electrolyte conductivity is only one of the factors that affect the blockade ionic currents. Figure 3 Scatter plot of the event residence time versus its blocked ionic current amplitude. In Figure 3, some outliers we call as ‘trapped events’ have

been observed in 1 M MgCl2 experiments. Although the probability is small, the duration time of these events is 22 ms, about PLEKHB2 17 times of the other events in 1 M MgCl2 experiments. As we know, Si3N4 surface in aqueous solution at pH 8.0 is negatively charged. The correlations between Mg2+ ions on both the negatively charged DNA and the Si3N4 surface can generate a net attraction force and then help stick the DNA into the nanopore, but the phenomenon only obviously occurred for the 7-nm diameter nanopore experiments. This is because the gap between the DNA and the inner surface of the nanopore is also increased with the increasing nanopore diameter. With the increase of the gap, the net attraction force is not strong enough to stick the DNA, which leads to the trapped events unremarkable in the 22-nm diameter nanopore. From Figure 3, we find not only the blockade current amplitude and duration time but also the event point dispersion degree increase with the increasing Mg2+ ion concentration.

Bacterial cultures were diluted in PBS to equal the McFarland No. 0.5 standard and the final inoculum Talazoparib chemical structure was prepared by diluting the bacterial suspension at 1:100. Aliquots

of 0.1 mL were transferred to each well of a 96-well plate that contained 0.1 mL of each compound at concentrations prepared from 2-fold serial dilutions in 7H9/OADC medium. The inoculated plates were incubated at 37°C until growth in the agent-free control-well was evident (2-3 days). The MIC was defined as the lowest concentration of compound that inhibited visible growth. Semi-automated fluorometric method The assessment of accumulation and extrusion of EtBr on a real-time basis by M. smegmatis strains wild-type mc2155, SMR5, porin mutants, MN01 and ML10 and efflux mutants XZL1675 and XZL1720

(Table 1) was performed using the semi-automated fluorometric method, as previously described [25–27]. (i) Accumulation assay M. smegmatis strains were grown in 5 mL of 7H9/OADC medium at 37°C until an O.D.600 of 0.8. Cultures were centrifuged at 13000 rpm for 3 minutes, the supernatant discarded and the pellet washed in PBS (pH 7.4). The O.D.600 was adjusted to 0.4 with PBS and glucose was added at final concentration of 0.4%. Aliquots of 0.095 mL of bacterial suspension were distributed to 0.2 mL PCR microtubes and EtBr was added at concentrations that ranged from 0.25 to 8 mg/L. Fluorescence was measured in the Rotor-Gene™ 3000 (Corbett Research, Sydney, Australia), selleck kinase inhibitor using the 530 nm band-pass and the 585 nm high-pass filters as the excitation and detection wavelengths, respectively. Fluorescence data was acquired every 60 seconds for 60 minutes at 37°C. The effect of chlorpromazine, thioridazine and verapamil on the accumulation of EtBr was determined by adding 0.005 mL of each compound to aliquots of 0.095 mL of EtBr-containing bacterial suspension previously distributed to 0.2 mL PCR microtubes. Fluorescence was measured every 60 seconds for 60 minutes at 37°C in the Rotor-Gene™ 3000. Each inhibitor was used at ½ the MIC in order to not compromise

the cellular viability (as Methocarbamol confirmed by CFUs counting). (ii) Efflux assay Mycobacteria were exposed to conditions that promote maximum accumulation of EtBr: EtBr at ½ MIC for each strain; no glucose; presence of the efflux inhibitor that caused maximum accumulation, in this case verapamil; and incubation at 25°C [25–27]. The EtBr loaded cells were centrifuged at 13000 rpm for 3 minutes and resuspended in EtBr-free PBS containing 0.4% glucose. After adjusting the O.D.600 to 0.4, aliquots of 0.095 mL were transferred to 0.2 mL microtubes. Fluorescence was measured in the Rotor-Gene™ 3000 as described for the accumulation assay. Efflux activity was quantified by comparing the fluorescence data obtained under conditions that promote efflux (presence of glucose and absence of efflux inhibitor) with the data from the control in which the mycobacteria are under conditions of no efflux (presence of an inhibitor and no energy source).

PCR-based prescreening for clones with DNA imports in strain 26695 uvrA Due to the low recombination frequency in 26695 uvrA, it was necessary to screen the Rif resistant clones after transformation in order to distinguish recombinants from spontaneous mutants. This was accomplished by allele-specific PCR using the primers HPrpoB-IscrX and HPrpoB-4, which specifically detect the Rif resistance mediating point mutation in strain J99-R3 [12, 46]. PCR positive clones were used for sequencing as described above. UV irradiation of mutant

strains Bacteria were cultured on blood agar plates for find more 24 h as described above. Cells were then suspended in phosphate buffered saline (PBS) and appropriate dilutions to obtain ~100, 500 and 1,000 colonies were plated on blood agar plates in two triplicate batches. As a control, the

first batch was not exposed to UV light to obtain the total cell number. The plates of the second batch were placed under a UV-C lamp (OSRAM HNS 30 W OFR, wavelength 254 nm) for two seconds at click here a distance of 40 cm, corresponding to approximately 100 J/m2. All plates were incubated for 72 h as previously described, colonies were counted and the percentage of surviving cells was calculated. Growth properties of H. pylori strains Growth curves were monitored in liquid cultures (BHI broth including 10% horse serum and antibiotics). Strains were grown for

<24 h on blood agar plates and then harvested in BHI broth. The OD600 of the suspension was measured and diluted to a starting concentration of 2.1 × 107 bacteria/ml. Cultures were then incubated at 37°C in a rotary shaker (175 rpm) under microaerobic conditions. The optical density was measured at regular intervals. Statistical methodology Statistical analysis was performed using Bayesian model comparison, where two competing hypotheses are weighted against each other by computing the ratio of probabilities of the observed data under the two hypotheses. This ratio is called a Bayes Factor (see refs. [47, 48] for reviews). A benefit of this approach is that it accounts for the relative Orotic acid complexity of the hypotheses, so that the more complex one is validated only if the data justifies it. Interpretation of the Bayes Factor was done following the scale of Jeffreys [49]: Negative (<1); Barely worth mentioning (1–3); Substantial (3–10); Strong (10–30); Very strong (30–100); Decisive (>100). When the Bayes Factor could not be analytically computed, the Bayes Information Criterion (BIC; refs. [47, 50] was used as an estimate: (1) where l 1 and l 2 are the maximized value of the log-likelihood under the two models, k 1 and k 2 the number of parameters in the two models, and n the number of observations.

01 0.02 6 Papuasia 2402 1 5 5 0.07 0.01 Biogeographical regions are sorted by ascending distance to the study area in Sulawesi. Probability (based on Poisson probability density) is related to the tree species pool observed in both studied sites (71 spp. assigned to valid species names) The likelihood analysis that one of the two

studied forest areas (N, R) included more tree species with nearest neighbour distance to one of the seven islands than the other were not significant, but showed some contrasting trends in biogeographical affinities of the two forest communities (Fig. 2). The mid-montane forests showed the greatest similarity with the western Malesian islands of Sundaland, especially Borneo, whilst the upper-montane forests had a great eastern Malesian affinity with New Guinea Z-VAD-FMK in vitro and also to the Philippines. Endemics to Sulawesi and to Maluku, i.e. Wallacean distributed species, were of equal importance at both sites. Fig. 2 Observed number of tree species GSK-3 signaling pathway (white squares) in the mid-montane forest at Mt Nokilalaki (42 spp.) and the upper montane forest at Mt Rorekautimbu (45 spp.) with nearest neighbour occurrences in seven Malesian biogeographical

regions, and expected patterns (black bars) based on 1000 random samples from the combined tree species pool (71 spp.). Biogeographical regions are sorted by ascending nearest neighbour distances (cf. Table 3) Discussion Elevational patterns in high mountain tree community

composition and structure The high mountain forests in Sulawesi show divergent patterns related to different elevational belts, both in floristic composition and in community GNAT2 dominance of certain taxa. In the Malesian mountain flora, within the montane zone sensu stricto (1600–2400 m a.s.l.), a major species shift indicates an orographic boundary at about 2000 m a.s.l. (van Steenis 1972). The present study supports these findings by showing a species shift between mid- and upper montane elevations (1800–2400 m a.s.l.), with only 18 species in common considering the total data set of 87 tree species (21%). Further, the mossy aspect of the forest at upper montane elevations (Gradstein and Culmsee 2010) also provides evidence for the elevational differences between the investigated forests. In the Fagaceae–Myrtaceae forests surveyed at mid-montane elevations, the Fagaceae play a key role. While four species of Lithocarpus contributed nearly half of the stand basal area, the importance of the family decreased at upper montane elevations in favour of the Podocarpaceae and Phyllocladaceae. Previous studies in Lore Lindu National Park, Central Sulawesi, showed that the Fagaceae were of comparable overall importance at lower montane elevations (at 1400 m a.s.l.), but became less important at submontane elevations (at 1050 m a.s.l.) (Culmsee and Pitopang 2009).

8Kb; hpdC 3.4 kb and for R20291: hpdA – 6.3 kb; hpdC – 3.4 kb. Analysis of the decarboxylase mutants Initial growth dynamics and NMR spectroscopy analysis revealed that the hpdB, hpdC and hpdA mutants were indistinguishable in terms of the complete lack of p-cresol production in rich media supplemented with p-HPA (Figure 4A). Subsequent analysis ZD1839 price was performed with the hpdC mutants as these were constructed in both parent strains R20291 and 630Δerm. Growth curves in minimal

media (YP broth) revealed that the R20291ΔhpdC mutant grew significantly better than the parent strain R20291, however, no significant difference in in-vitro growth was observed between 630ΔermΔhpdC and the respective parent strain (Figure 4B). There were no significant selleck inhibitor differences between the tolerance of the mutants R20291ΔhpdC and 630ΔermΔhpdC to 0.1% p-cresol compared

to their respective parent strains (Figure 4C), however, the R20291 strains (wild-type and R20291ΔhpdC) are significantly more tolerant to p-cresol than their 630 counterparts (wild-type and 630ΔermΔhpdC) (p < 0.01). The absence of p-cresol production observed in the R20291ΔhpdC and 630ΔermΔhpdC mutants by NMR spectroscopy in rich media supplemented with 0.1% p-HPA (Figure 4A), was reproducible in minimal media using zNose™ gas chromatography (data not shown). Figure 4 Analysis of the decarboxylase mutants. A) NMR spectra showing p-cresol production in BHI broth supplemented with 0.1% p-HPA for parent and mutant strains, B) Growth curve of the R20291ΔhpdC and 630ΔhpdC mutants compared to respective parent strains. C) Tolerance to 0.1% p-cresol of ΔhpdC mutants and respective parent strains. Temporal production of p-HPA and p-cresol in mutant and parent strains Preliminary NMR spectroscopy revealed that p-cresol was produced in unsupplemented minimal media

(YP broth), indicating that the available tyrosine was converted to p-cresol via the intermediate p-HPA. The temporal production of p-HPA and p-cresol were assessed in minimal YP media, using both wild-type and mutant strains of R20291 and 630Δerm. For each strain, samples were taken every hour for the first 8 hours with a final time point of 24 Protein tyrosine phosphatase hours, after which the relative production of p-HPA and p-cresol were determined by NMR spectroscopy, the combined data for all the strains and controls is presented in Figure 5A. High levels of tyrosine were present in all samples including the media control (Figure 5A); however, the conversion to p-HPA and p-cresol across all the strains was limited to a few samples (Figure 5A), namely the latter time points in the parent strains. In the decarboxylase mutants R20291ΔhpdC and 630ΔermΔhpdC, a build up of p-HPA was evident from 4 to 24 hours (Figure 5B and 5C). The level of p-HPA production was significantly higher in the R20291ΔhpdC mutant compared to the 630ΔermΔhpdC mutant (Figure 5B and 5C). As predicted, p-cresol was not detected in the mutant samples.

76% >0.99 Saccharomycotina Pichia ohmeri 102.31% >0.99 Saccharomycotina Saccharomycopsis crataegensis 80.98% >0.99 Saccharomycotina Stephanoascus ciferrii 85.84% >0.99 Mucoromycotina Absidia corymbifera 92.33% >0.99 Mucoromycotina Cunninghamella

bertholletiae 80.03% >0.99 Mucoromycotina Rhizopus microsporus 89.16% >0.99 Mucoromycotina Rhizopus oryzae 87.96% >0.99 Pezizomycotina Alternaria sp. 103.70% >0.99 Pezizomycotina Cladosporium cladosporioides 92.87% >0.99 Pezizomycotina Cytospora chrysosperma 100.50% >0.99 Pezizomycotina Endoconidioma sp. 89.93% >0.99 Pezizomycotina Geopora sp. 114.45% >0.99 Pezizomycotina Phoma herbarum 91.94% >0.99 Pezizomycotina Xanthomendoza galericulata 94.27% >0.99 Agaricomycotina Agaricus sp. 95.31% >0.99 Agaricomycotina Clavulina coralloides 99.59% >0.99 Selleckchem SB203580 Agaricomycotina Coprinus sp. 99.70% >0.99 Agaricomycotina Cortinarius sp. 102.68% >0.99 Agaricomycotina Hebeloma crustuliniforme group 91.06% >0.99 Agaricomycotina Melanogaster sp. 102.27% >0.99 Agaricomycotina Pleurotus ostreatus 102.71% >0.99 Agaricomycotina Rhizopogon sp. 107.04% >0.99 Agaricomycotina Sclerogaster xerophilus 92.17% >0.99

Agaricomycotina Sedecula pulvinata 92.26% >0.99 Agaricomycotina Tricholoma populinum 89.53% >0.99 Agaricomycotina Trichosporon asahii 78.03% >0.99 Agaricomycotina Trichosporon asteroides 82.66% >0.99 Agaricomycotina Trichosporon cutaneum 86.66% >0.99 Agaricomycotina Trichosporon LDE225 nmr dermatis 80.27% >0.99 Agaricomycotina Trichosporon faecale 84.05% >0.99 Agaricomycotina Trichosporon montevideense 77.43% >0.99 Agaricomycotina Trichosporon mucoides 82.87% >0.99 Agaricomycotina Trichosporon ovoides 105.59% >0.99 Pucciniomycotina Rhodotorula mucilaginosa 96.29% >0.99 Pucciniomycotina Rhodotorula slooffiae 99.94% >0.99 Agaricomycotina Lactarius sp. 86.76-89.03% >0.99 Bay 11-7085 Table 4 FungiQuant quantitative validation

results, obtained using pure plasmid standards and different mixed templates Templates tested Assay quantitative dynamic range Average reaction efficiency (SD) r 2 -value 10 μl Reaction       Plasmid standards-only 25 – 107 copies 91.80% (1.91%) >0.99 Plasmid standards plus 0.5 ng human DNA 25 – 107 copies 93.20% (0.70%) >0.99 Plasmid standards plus 1 ng human DNA 25 – 107 copies 97.02% (4.97%) >0.99 Plasmid standards plus 5 ng human DNA 25 – 107 copies 92.85% (1.33%) >0.99 Plasmid standards plus 10 ng human DNA 25 – 107 copies 91.21% (1.79%) >0.99 C. albicans DNA-only 10 fg – 10 ng 94.75% (2.33%) >0.98 C. albicans DNA plus 1 ng human DNA 10 fg – 10 ng 96.84% (1.93%) >0.99 5 μl Reaction       Plasmid standards-only 25 – 107 copies 92.17% (5.64%) >0.98 Plasmid standards plus 1 ng human DNA 25 – 107 copies 94.21% (2.92%) >0.99 Plasmid standards plus 10 ng human DNA 50 – 108 copies 92.64% (2.39%) >0.99 Table 5 Interpretation of FungiQuant results for detecting fungal DNA (i.e.