We and others have previously reported

that certain determinants of EPEC, which are not encoded by LEE or the EAF plasmid, such as Efa1, NleB and the cytolethal distending toxin (Cdt) are associated with virulence in attaching-effacing E. coli [15, 27]. NleB was detected in 20 (30%) of the 67 strains tested, whereas Efa1 was detected in 8 strains (12%), all of which were also positive for NleB. NleB-positive strains were distributed amongst the following clades: EHEC-1 (3 strains), EPEC-2 (2 strains), aEPEC-1 (intimin-β, H7; 3 strains), aEPEC-3 (intimin-θ, H21 [or H6]; 6 strains). Six NleB-positive isolates could not be assigned to a clade, although all carried intimin-θ (Table 1). The Efa1-positive strains occurred within the EHEC-1 and EPEC-2 clades, as well as within the aEPEC-1 clade that was characterised by

strains with intimin-β and ASP2215 price AG-881 mw H7. Seven (10%) strains were positive in the PCR for Cdt. Three of these strains belonged to the aEPEC-6 clade (intimin-α and H34), one belonged to EPEC-2 (intimin-β, H2), and three were unassigned (Table 1). DNA hybridization To determine if aEPEC carry DNA sequences related to those that code for the production of BFP, but were not amplified by the PCR for BfpA, we investigated the aEPEC strains by DNA learn more hybridisation using probes derived from the bfpA and bfpB genes of EPEC strain E2348/69. Unexpectedly, six isolates (ESA212, R176, R182, R228-1, R281, and W114) hybridised with the BfpA probe at high stringency. Three of these strains belonged to the aEPEC clade with intimin-ι and H8, but they belonged to different O-serogroups. The other three probe-positive strains also differed from each other. Six strains hybridised with the BfpB probe. Four of these were positive for intimin-α, three carried H34, two carried H6, but all were of different serotypes. No strain

hybridised with both Bfp probes. Some aEPEC strains from humans and animals express adhesins that are homologous to the K88 fimbriae of enterotoxigenic E. coli [21, 28]. To determine if the aEPEC strains in our collection carried similar sequences, we probed these strains for the fae gene of K88, but none of the aEPEC N-acetylglucosamine-1-phosphate transferase hybridised with this probe, even when tested at low stringency. Adherence to HEp-2 cells After incubation for three hours with HEp-2 cells, 54 (81%) of 67 aEPEC strains were adherent: 24 strains adhered in an aggregative pattern, and two in the pattern termed “”localised-like adherence”", because it resembles BFP-mediated localised adherence, but the bacteria are more loosely associated with each other than BFP-bearing strains. Twenty-eight strains showed an indeterminate pattern of adherence described previously [20], in which bacteria adhere in a mixed pattern of diffuse and localised-like adherence. Thirteen strains did not adhere to HEp-2 cells after 3 hours.

Phys Rev B 1972, 6:4370–4379.CrossRef 40. Kabashin AV, Evans P, Pastkovsky S, Hendren W, Wurtz GA, Atkinson R, Pollard R, Podolshiy VA, Zayats AV: Plasmonc nanorod metamaterials for biosensing. Nat. Mater. 2009, 8:867–871.CrossRef 41. Wurtz G, Pollard R, Hendren W, Wiederrecht G, Gosztola D, Podolskiy V, Zayats A: Designed ultrafast optical nonlinearity in a plasmonic nanorod

metamaterial enhanced by nonlocality. Nat Nanotechnol 2011, 6:106–110.CrossRef 42. Pollard R, Murphy A, Hendren W, Evans P, Atkinson R, Wurtz G, Zayats A: Optical nonlocalities and additional waves in epsilon-near-zero metamaterials. Phys Rev Lett 2009, 102:127405.CrossRef 43. Nielsch K, Müller F, Li AP, Gösele U: Uniform nickel deposition into ordered OSI-906 order alumina pores by pulsed electrodeposition. Adv Mater 2000, 12:582–586.CrossRef 44. Novotny L, Hecht B: Principles of Nano-optics. Cambridge: Nirogacestat Cambridge University Press; 2006.CrossRef 45. Wang QQ, Han JB, Guo DL, Xiao S, Han YB, Gong HM, Zou XW: Highly efficient avalanche multiphoton luminescence from coupled Au nanowires in the visible region. Nano Lett 2007, 7:723–728.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions

JX, ZKZ, ZL, and JY prepared the samples. JX, QZ, and ZKZ anticipated the optical experiments and analyzed the related experiment data. JX, ZKZ, and YL characterized the morphology of the samples. JX https://www.selleckchem.com/products/isrib-trans-isomer.html and ZKZ performed the simulations using FDTD solution and interpreted the simulation results. JML, JTL, and XHW performed the numerical simulation of Dapagliflozin the LDOS section. ZKZ proposed the pulse AC growth method and finalized the manuscript. All authors read and approved the final manuscript.”
“Background The rapid proliferation of advanced electronic devices for many commercial and military applications, such as data transmission, telecommunications,

wireless network systems, and satellite broadcasting as well as radar and diagnostic and detection systems, has led to numerous electromagnetic compatibility and electromagnetic interference (EMI) problems. The interaction of electromagnetic waves originating from different sources can lead to a decrease in quality and a misinterpretation of transferred data, and it has thus become vital to avoid such interference and electromagnetic wave pollution through the use of appropriate absorbing and shielding materials. Carbonaceous materials – such as graphite and/or carbon black – are often used as dielectric electromagnetic absorbers, generating dielectric loss by improving the electrical conductivity of the mixture. In particular, nanostructured materials and carbon fiber composites have been the subjects of growing interest as microwave radiation absorbing and shielding materials in the high-frequency range due to their fascinating properties [1–5].

Interestingly, the changes

in the levels of Kid/KIF22 mRNA mirrored that of SIAH-1 in all of the patients. Kid/KIF22 mRNA levels were decreased in all tumors in which SIAH-1 mRNA was decreased and vice versa (Figure 4). Moreover, except for one sample, the number of Kid/KIF22 mRNA copies was consistently higher than the SIAH-1 mRNA copies in all normal tissues (with a median of 19,2 × 103) compared to their corresponding paired tumor tissues (median of 16,5 × 103). Discussion In this study, we compared SIAH-1 mRNA and protein expression levels in normal and tumor tissues and cell lines. SIAH-1 protein was found to be widely expressed in human cell lines and tissues. In non-proliferating tissues that express higher levels of SIAH-1 mRNA, a single band of the expected MW is detectable (muscle), or it represents almost the

totality of the detected protein 3-deazaneplanocin A (brain). In other tissues and majority of cells lines a second band appears whose molecular EPZ5676 weight is approximately the double of the first one. PRIMA-1MET manufacturer Although it is known that SIAH-1 forms stable homodimers [2, 3, 29], under reducing conditions used in SDS-PAGE a single band would be expected. The additional bands observed in Figure 1 could correspond to post-translational modifications, or to transcriptional or splicing variants of SIAH-1. Indeed, human SIAH-1 mRNA is 2.3 kb but an additional transcript of 2.5 kb was shown in placenta [5]; in MCF-7 cells, a SIAH-1 variant that encodes a 298 selleck chemical amino acid protein designated SIAH-1L was reported [30] whereas another variant named SIAH-1S encoding a 195 amino acid protein

was detected in breast, Kidney and esophagus cancer tissues [31]. The broad tissue distribution of SIAH-1 suggests that it may play a relevant cellular role; however, high levels and splicing variants of SIAH-1 in particular tissues may represent sites of critical gene function or relate to physiological/pathological situations. Consistent with this, important differences in SIAH-1 expression were observed amongst cell lines and tissues. Interestingly, in some tissues such as the small intestine, other bands of high molecular weight appear suggesting the presence of polyubiquitinated forms of SIAH-1. This observation is consistent with previous reports, since SIAH-1 was shown to be auto-ubiquitinated and degraded via the proteasome pathway [2, 3] and we showed a strong SIAH-1 expression in the cells at the apical of the intestine villi, where cells are differentiated and die by apoptosis [17]. By fluorescence microscopy, SIAH-1 was shown to be highly expressed in the cytoplasm of normal breast cells, with a punctuate pattern. In tumor tissues however, it appeared as a more uniform distribution, localized to both the cytoplasm and nucleus. Similarly, whereas in normal liver the expression was high and homogeneous among cells, tumor tissues showed significant heterogeneity with some cells expressing high levels of SIAH whilst being undetectable in others.

For these drugs the employ of intravenous continuous infusion, which ensures the highest steady-state concentration under the same total daily dosage, may be the most effective way of maximizing pharmacodynamic exposure [51–54]. On the other hand, quinolones, daptomycin, tigecycline, aminoglycosides, polienes and echionocandins exhibit concentration-dependent activity; therefore the entire daily dose should be administered in a once daily way (or Apoptosis inhibitor with the lowest possible number of daily administrations) with the intent of achieving the highest

peak plasma level. The use of extended-interval aminoglycoside dosing strategies for the treatment of moderate-to-severe infections encountered in critically ill surgical patients [55, 56]. Classifications Intra-abdominal infections (IAIs) include a lot of pathological conditions, ranging from uncomplicated appendicitis to faecal peritonitis. From a clinical viewpoint IAIs are classified into uncomplicated and complicated [57]. GSK1210151A In uncomplicated IAIs the infectious process only involves a single organ and does not proceed to the peritoneum. In complicated IAIs, the infectious process proceeds beyond the organ, and causes either localized Selleck GSK2118436 peritonitis (intra-abdominal abscess), or diffuse peritonitis. Peritonitis is classified into primary, secondary or tertiary peritonitis [58].

Primary peritonitis is a diffused bacterial infection without loss of integrity of the gastrointestinal tract. It is rare. It mainly occurs in infancy and early childhood heptaminol and in cirrhotic patients. Secondary peritonitis, the most common form of peritonitis, is acute peritoneal infection resulting from loss of integrity of the gastrointestinal tract or from infected viscera. It is caused by perforation of the gastrointestinal tract (e.g.

perforated duodenal ulcer), by direct invasion from infected intra-abdominal viscera (e.g. gangrenous appendicitis). Anastomotic dehiscences are common causes of peritonitis in the postoperative period. Tertiary peritonitis is defined as peritonitis that persists after more than one failed source control procedure [59]. Intra-abdominal infections are also classified into community-acquired intra-abdominal infections (CA-IAIs) and healthcare-acquired intra-abdominal infections (HA-IAIs). CA-IAIs are acquired in community, whereas HA-IAIs develop in hospitalized patients or residents of long-term care facilities. They are characterized by increased mortality because of both underlying patient health status and increased likelihood of infection caused by multi drugs resistant organisms [59]. Moreover, in the classification of IAIs should be mandatory to introduce a grading of clinical severity, well represented by the sepsis definitions. The updated sepsis definition is based on several clinical and bioumoral variables [60].

Ears: hearing loss (Alport syndrome, adverse effects of aminoglycoside antibiotics). Oral cavity: macroglossia (amyloidosis), tonsillar hypertrophy, fur (IgA

nephropathy, streptococcal infection), cervical vein dilatation, collapse (assessment of body fluid), bruit over the neck (atherosclerosis). Chest: Lazertinib clinical trial signs of heart failure (heart murmurs, pulmonary edema, pleural fluid), pulmonary alveolar hemorrhage, Selleckchem BIX 1294 epicarditis (SLE, uremia). Abdomen: bruit (renal artery stenosis), palpable kidney (polycystic kidney), tap pain over the kidney (acute pyelonephritis, renal infarction), abdominal pain (Henoch–Schönlein purpura, cholesterol embolus). Prostate gland: hypertrophy (urinary obstruction, post-renal acute renal failure). Extremities: edema (body fluid retention), arthralgia

or joint deformity (gout, rheumatoid arthritis, collagen disease, Henoch–Schönlein purpura), blue toe (cholesterol embolus), pains (Fabry disease). Skin: poor turgor (dehydration), purpura AC220 cell line (Henoch–Schönlein purpura), livedo reticularis (reticular rash: cholesterol embolus, vasculitis), angiokeratoma/acroparesthesia/anhidrosis (Fabry disease).”
“It is important in the follow-up of CKD patients to slow worsening of the disease and to prevent CVD. In the case of eGFR ≥ 50 mL/min/1.73 m 2 , primary care physicians manage CKD, collaborating with nephrologists. In the case of eGFR < 50 mL/min/1.73 m 2 , primary care physicians and nephrologists manage CKD concurrently. A patient is recommended to be referred to nephrologists

immediately after onset of abrupt increase of urinary protein or rapid decline of eGFR. Strategies of follow-up vary depending on primary diseases for CKD. Urinalysis, calculation of eGFR, and image testing are conducted at regular intervals to assess kidney function as well as to try to find CVD. Reasons for importance of CKD follow-up Progression of each CKD stage toward end-stage kidney disease (ESKD) is accelerated as the stage advances. It is therefore necessary to confirm therapeutic effectiveness in order to slow CKD progression. Even in stages 1–3, the probability of death from cardiovascular disease (CVD) is greater than that of proceeding to ESKD. It is possible to slow the progression of CKD by lifestyle education and drug therapy, Oxaprozin but regular follow-up is required to determine their efficacy. It has been evidenced that control of blood glucose as well as blood pressure and use of ACE inhibitors as well as ARBs is effective in suppressing CKD progression. Treatment of dyslipidemia or anemia or restriction of dietary protein also has similar effects. Follow-up differences depend on primary diseases Diabetic CKD has a high prevalence of CVD and progresses rapidly in kidney function. Blood glucose should be controlled to keep HbA1c below 6.5%. ECG and cardiac echography are recorded to prevent CVD development.

To confirm these observations, we performed quantitative analyses using the XTT assay. Figure 4A shows that after 2 days of culture, KSL-W was able to inhibit biofilm formation. This inhibitory effect was observed beginning at 25 μg/ml of KSL-W. At concentrations of 50, 75, and 100 μg/ml of KSL-W, the inhibition of C. see more albicans biofilm formation was comparable to that caused by amphotericin B at 10 μg/ml.

Similar results were obtained after 4 days (Figure 4B) and 6 days (Figure 4C) of culture for biofilm formation with a persistent inhibitory PLX4032 cell line effect of KSL-W on C. albicans biofilm formation. Figure 3 Scanning electron microscope analyses of the biofilm formation. C. albicans was cultured in Sabouraud medium with or without KSL-W at various concentrations for 4 days in a porous 3D collagen scaffold. Cultures in the presence of amphotericin B (10 μg/ml) were used as the positive controls. Following incubation, the samples were prepared as described in the Methods section and were

observed Tozasertib cell line under a scanning electron microscope. Negative control refers to the non-seeded scaffolds. Figure 4 Quantitative measurement of the reduced biofilm formation with KSL-W. C. albicans was cultured on a 3D porous scaffold in the presence of KSL-W for 2, 4, and 6 days. After each culture period, the samples were supplemented with XTT solution and incubated for 5 h at 37°C. The absorbance at 450 nm was measured to quantify XTT metabolic product intensity proportional to the number of viable cells. (A) 2 days; (B) 4 days; (C) 6 days. Results are means ± SD for three different Dichloromethane dehalogenase separate experiments. KSL-W disrupted mature C. albicans biofilms After 6 days of incubation in glucose-rich Sabouraud medium, scaffolds seeded with C. albicans strain SC5314 produced mature biofilms displaying highly dense populations of Candida cells (Figure 5). Significant reductions and disruptions of the pre-formed Candida biofilms were observed when the reference antifungal agent (amphotericin B, 10 μg/ml) was added to the mature biofilms upon further incubation up to 6 days. Similarly, antimicrobial peptide KSL-W at 75 and 100 μg/ml also

reduced C. albicans density in the biofilms. The observed reduction was noticed with KSL-W concentrations ranging from 25 to 100 μg/ml. Indeed, when quantitatively investigated by XTT reduction assay, the KSL-W-treated biofilms rendered a significantly lower number of cells, as reflected by the lower absorbance readings, than did the untreated control. This effect was observed after 2, 4, and 6 days of treatment with amphotericin B. Furthermore, the effect of KSL-W on the mature C. albicans biofilm was comparable to that obtained with amphotericin B (Figure 6). Figure 5 Biofilm ultrastructure following KSL-W treatment. C. albicans was cultured in Sabouraud medium without KSL-W for 6 days to promote biofilm formation and maturation.

01 Saliva   Ca [mg/L] 61.691 ± 36.851 70.771 ± 57.572 NS   Zn [mg/L] 2.043 ± 1.511 2.652 ± 1.792 NS   Cu [μg/L] 114.644 ± 78.362 78.321 ± 61.691 NS Serum   Ca++ ionized (mmol/L) 1.21 ± 0.07 1.20 ± 0.06 NS   Ca (mmol/l) 2.36 ± 0.10 2.36 ± 0.11 NS   Zn [mg/L] 1.042 ± 0.242 1.161 ± 0.222 NS   Cu [mg/L] 0.741 ± 0.205 0.713 ± 0.212 NS   Phosphate (mg/dL) 3.16 ± 0.51 3.08 ± 0.55 NS   PTH

(pg/mL) 58.69 ± 28.31 56.56 ± 22.04 NS   25(OH)D (ng/mL) 24.33 ± 6.29 22.19 ± 5.63 NS   Alkaline phosphatase (sALP) [IU/L] 55.14 ± 15.81 62.35 ± 17.59 NS   Osteocalcin (ng/mL) 19.87 ± 6.05 18.75 ± 4.62 NS NS denote not statistically significant differences Discussion Our study showed coincidence of reduced spine PF-01367338 BMD and local enamel copper deficits in a group of patients suffering from a rare disorder: advanced tooth wear. This association was independent of dietary intake of copper or serum content of this element either. Some properties of saliva are considered an important biological factor affecting the rate of dental erosion, transporting ions, and mineralization balance [44, 45]; however, the low enamel copper in our patients was unaffected by salivary concentrations of this trace element. Considering

other variables studied, i.e., bone formation markers, PTH, vitamin D status, or menopausal status in women, the decreased Dehydrogenase inhibitor copper concentration may potentially play a role in the pathophysiology of mineralized tissues in human body. Whether there is a casual link between the Cu depletion in situ,

susceptibility to lower BMD and advanced tooth abrasion, remains not fully understood. Several studies in adult populations have documented associations between systemic bone loss in the development of tooth loss [4, 5, 7, 17, 46]. Resorption of tooth-supporting alveolar bone has been regarded as one of the responsible mechanisms [2, 17, 20]. Nonetheless, available data are inconsistent, and usually based on a self-reported tooth count. Most studies have focused on postmenopausal women, but a few reports have shown lower BMD being associated with the number of missing teeth also in men [12, 47]. Whereas certain studies in edentulous elder population have shown reduced BMD at the spine accompanied by higher BMD at the femoral neck [48], others have reported contrasting findings, i.e., lower Clomifene femoral rather than spine BMD being associated with tooth loss [19]. These teeth–bones relationships relate usually to older people and prove clinical relevance of dental status in postmenopausal osteoporosis. We extend this observation by demonstrating that the onset of dental disease with precocious rapid enamel abrasion in younger age may also coexist with decreased BMD. The DXA AZD1480 research buy measurement, used in our study, does not allow insight into the structure or quality of bone, so that it is neither able to distinguish between cortical and trabecular bone loss nor between mineral and matrix deterioration.

J Appl Microbiol 2010,109(3):808–817.PubMedCrossRef 49. Olier M, Pierre F, Rousseaux S, Lemaitre JP, Rousset A, Piveteau P, Guzzo J: Expression of truncated Internalin A is involved in impaired internalization of some Listeria monocytogenes isolates carried asymptomatically by humans. Infect Immun 2003,71(3):1217–1224.PubMedCrossRef 50. Kim H, Bhunia AK: SEL, a selective AC220 enrichment broth for simultaneous growth of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes. Appl Environ Microbiol 2008,74(15):4853–4866.PubMedCrossRef 51. Walcher G, Stessl B, Wagner M, Eichenseher F, Loessner MJ, Hein I: Evaluation of paramagnetic

beads coated Tubastatin A with recombinant Listeria phage endolysine derived cell-wall-binding domain proteins for separation of Listeria monocytogenes from raw milk in combination

with culture-based and real-time polymerase chain reaction based quantification. Foodborne Pathog Dis 2010,7(9):1019–1024.PubMedCrossRef 52. Paoli GC, Kleina LG, Brewster JD: Development of Listeria monocytogenes-specific H 89 immunomagnetic beads using a single-chain antibody fragment. Foodborne Pathog Dis 2007,4(1):74–83.PubMedCrossRef 53. Tully E, Hearty S, Leonard P, O’Kennedy R: The development of rapid fluorescence-based immunoassays, using quantum dot-labeled antibodies for the detection of Listeria monocytogenes cell surface proteins. Int J Biol Macromol 2006,39(1–3):127–134.PubMedCrossRef 54. Bueno VF, Banerjee P, Banada PP, de Jose MA, Lemes-Marques EG, Bhunia AK: Characterization of Listeria monocytogenes isolates of food and human origins from Brazil using molecular typing procedures and in vitro cell culture assays. Int J Environ Health Res 2010,20(1):43–59.PubMedCrossRef 55. Jacquet C, Doumith M, Gordon JI, Martin PM, Cossart P, Lecuit M: A molecular marker for evaluating the pathogenic potential of foodborne Listeria monocytogenes. J Infect Dis 2004,189(11):2094–2100.PubMedCrossRef 56. Chen Y,

Ross WH, Whiting RC, Van SA, Nightingale KK, Wiedmann M, Scott VN: Variation in Listeria monocytogenes dose responses in relation to subtypes encoding a full-length or truncated internalin A. Appl Environ Microbiol 2011,77(4):1171–1180.PubMedCrossRef Ponatinib mouse 57. Varshney M, Yang LJ, Su XL, Li YB: Magnetic nanoparticle-antibody conjugates for the separation of Escherichia coli O157:H7 in ground beef. J Food Protect 2005,68(9):1804–1811. 58. Foddai A, Elliott CT, Grant IR: Maximizing capture efficiency and specificity of magnetic separation for Mycobacterium avium subsp. paratuberculosis cells. Appl Environ Microbiol 2010,76(22):7550–7558.PubMedCrossRef 59. Snapir YM, Vaisbein E, Nassar F: Low virulence but potentially fatal outcome – Listeria ivanovii. Eur J Intern Med 2006,17(4):286–287.PubMedCrossRef 60.

Methods Isolate characterization Isolates were originally obtained during the large waterborne outbreak of C. jejuni and E. coli O157:H7 in Walkerton, Ontario in 2000. Strain typing was done previously [22]. All four human clinical isolates were epidemiologically related as part of a large water-borne outbreak of Campylobacter in Ontario, Canada, in the year 2000 [22, 23]. The isolates were also very closely related by phenotypic and genotypic typing tests (Table 7). Other than PFGE restriction profile, which we have previously shown resulted from movement of the prophage in the chromosomes [3], the only Selleck OICR-9429 difference was that isolate 00–2544 was PT35 rather than PT33. Table 7 Characteristics

of clinical C. jejuni isolates selleck screening library used for adherence

and invasion assays (from Clark et al . [[22],[23]]) Isolate Bio type ST flaA SVR type fla-RFLP HS serotype HL serotype https://www.selleckchem.com/products/3-deazaneplanocin-a-dznep.html Phage type PFGE Sma I PFGE Kpn I 00-2425 II 21 36 1 O:2 125 33 2 2 00-2426 II 21 36 1 O:2 125 33 1 1 00-2538 II 21 36 1 O:2 125 33 11 1 00-2544 II 21 36 1 O:2 125 35 4 1 ST, sequence type according to the Oxford MLST scheme; flaA SVR, sequence of the flagellin short variable region DNA sequence according to the Oxford scheme; fla-RFLP, restriction fragment-length polymorphism of the amplified complete flaA locus; HS, heat-stable or Penner serotype; HL, heat-labile or Lior serotype. Isolates 00–2425, 00–2538, and 00–2544 all carried a prophage homologous to CMLP1 (CJIE1) of strain RM1221. Isolate 00–2426 lacked this prophage. The motility of each isolate was assessed by applying 10 μl of growth from Brucella broth adjusted to OD600 = 0.1 onto semi-solid agar (Oxoid Mueller-Hinton broth + 0.4% Select Agar). Zones of motility were measured after growth for 48 h at 37°C under microaerobic conditions. Growth curves Bacteria grown on Oxoid

Mueller Hinton Agar + 10% sheep erythrocytes were used to inoculate 50 ml BBLTM Brucella Broth Albimi (VWR Canada, Mississauga, ON, Canada). After overnight growth, each suspension was diluted to an OD600 of approximately 0.18 Glutamate dehydrogenase to 0.2 (approximately 2 × 108 cfu/ml). This suspension was diluted by 10-4 to a concentration of approximately 2 × 104 cfu/ml and 0.5 ml of the resulting suspension was inoculated into 50 ml Brucella Broth Albimi to give approximately 200–500 cfu/ml. Growth proceeded for four days at 37°C under microaerobic conditions (5% O2, 10% CO2, 85% N2). At intervals aliquots were taken, diluted appropriately, and plated in duplicate onto Mueller-Hinton agar plates for determining viable cell counts. All plates with 20 – 300 colonies were counted, so that between 2 and 4 values were available for calculating the mean plus standard deviation of the cell concentration. Inoculated plates were incubated in microaerobic conditions at 42°C for 36 – 48 h, or at 37°C for 3 days, and colonies were counted. Data were plotted in Sigma Plot 10.0.1 (Systat Software Inc, San Jose, CA).

sp. URa15—Hochtor; Trebouxia sp. URa8—abernas;

T. sp. URa12—Gynge Alvar; T. sp. URa13—Hochtor). Table 4 Overview of chlorobiont occurrence in the four SCIN habitats   Genus Tabernas/Spain Hochtor/Austria Ruine Homburg/ Germany Gynge/Sweden Clades/ species Asterochloris sp. – 2 3 2 Chloroidium saccharophilum – 1 – – Trebouxia sp. 4 5 5 5 Other EGMA – 4 7 2 Other EGMA other eukaryotic green micro algae The key lichen P. decipiens occurred not only at all SCIN habitats but also in all additional soil crust specimens from other high Alpine areas. In most cases each individual lichen specimen contained one or more photobionts from every clade together with other eukaryotic green micro algae (EGMA; see Online Resource 1). The species specificity of the mycobiont towards its photobiont was quite low for P. decipiens. In contrast, Fulgensia bracteata ssp. deformis (which has so far only been found in samples from Hochtor) only occurred GSK461364 datasheet with T. sp. URa4 and A. sp. URa15 (the latter until now only known from this area, Figs. 2, 3). Peltigera rufescens, known to have a cyanobacterium as its primary photobiont (O’Brien et al. 2005), was also found to be associated with chlorobionts (Henskens et al. 2012). Specimens of P. rufescens from Ruine Homburg were associated with T. sp. URa6 and A. sp. URa16, although other

chlorobionts were available at the site; at Hochtor P. rufescens was found with T. impressa (see Online Resource 1, Figs. 2, 3). Discussion This evaluation of European lichen-dominated soil crusts from four geographically and climatically diverse sites revealed an unexpectedly high diversity of photobionts CHIR98014 supplier in association with the dominant lichen P. decipiens. Until now, only the genus Asterochloris has been described as the photobiont of P. decipiens (Schaper and Ott 2003), but we detected 12 different groups of the genus Trebouxia Acyl CoA dehydrogenase as well as other eukaryotic green micro algae like C. saccharophilum. Several of these micro algae are already known to exist as lichen photobionts, such as T. impressa, T. asymmetrica or the, as yet undescribed, Trebouxia sp. URa2, URa4, URa6.

The latter three species have also been identified as photobionts from crustose lichens (Ruprecht et al. 2012). Other Trebouxia species that are known as free-living algae (e.g. T. arboricola; Ettl and Gärtner 1995) were included in the analysis but not found in the soil-crust samples. P. decipiens at Hochtor showed a shared use of the available photobionts with other lichen species that were present (see Online Resource 1) with each species having a different level of specificity towards to its photobiont. We can conclude for P. decipiens that this lichen is not limited to a Ruxolitinib molecular weight single species or even genus of photobiont but instead associates with a broad range of apparently locally available algae. The low specificity of P.