Then it was centrifuged at 12,000 rpm for 30 min at 4°C The supe

Then it was centrifuged at 12,000 rpm for 30 min at 4°C. The supernatant was collected and stored at −80°C until use. The Antimicrobial activity of the supernatant was tested against C. albicans MTCC 3958, P. aeruginosa MTCC 741, S. aureus MTCC 737. Physicochemical properties of the anti-Candida compound Sensitivity to heat, pH,

and hydrolyzing enzymes Temperature stability was evaluated by incubating the CFS at various temperatures: 60°C for 90 see more min, 90°C for 20 min, 100°C for 20 and 30 min or autoclaved. Residual anti-Candida activity was determined by a well-diffusion assay against C. albicans. The effect of pH was determined using a pH range from 2 to 10 adjusted with diluted HCl or NaOH. After incubation at 37°C for 1 h, the resulting CFS was subjected to an agar-well diffusion assay to record the loss or retention of biological activity. Resistance to several proteolytic enzymes was tested by incubating the dialysed concentrate with pepsin, α-amylase, pronase E, trypsin, lipase and proteinase K at a final concentration of 1.0 mg mL-1. Buffers were used as controls. Samples were incubated at 37°C for

90 min. The residual activity was determined by cut-well agar assay. Effect of organic solvents, surfactants, and storage The sensitivity of dialyzed concentrate of ACP was tested in the presence of several organic solvents (methanol, ethanol, isopropanol, hexane, formaldehyde, chloroform, acetone and acetonitrile) at a final concentration of 25% (v/v). After incubation for 2 h at 37°C, the OSI-906 purchase Protein tyrosine phosphatase organic solvent was evaporated using a speed vac system (Martin Christ), and the residual antimicrobial

activity was determined. An untreated dialysed concentrate sample was taken as control. The effect of various surfactants, including Triton X-100, Tween-20, SDS, urea, EDTA, PMSF, and DTT (1.0% each) on the dialyzed concentrate was also tested. To assess whether the antifungal activity was due to the oxidation state of cysteine residues, β-mercaptoethanol (1 and 2 mmol) was used. The heat-treatment at 80°C was given for 10 min. In order to determine the stability, the CFS, dialyzed concentrate and partially purified ACP samples were stored for 1 year at low temperatures (4, −20 and −80°C) and the antimicrobial activity was compared to the freshly purified preparation. Partial purification of the anti-Candida compounds E. faecalis was cultured in mTSB medium at 14°C for 48 h. Cells were harvested by centrifugation at 12,000 rpm for 30 min at 4°C, and the CFS was filtered through 0.45 μm membranes. The culture supernatant was subjected to sequential ammonium sulphate precipitation to achieve 30%, 50% and 85% saturation at 4°C with constant and gentle stirring for 1 h. The precipitated proteins were pelleted by centrifugation at 12,000 rpm for 30 min. The protein pellet was dissolved in sterile 20 mmol sodium phosphate buffer pH 8.

Sequence similarities from Genbank BLASTn (XLSX 10 KB) Reference

Sequence similarities from Genbank BLASTn. (XLSX 10 KB) References 1. Ovreas L, Curtis TP: Microbial diversity and ecology. In Biological Diversity: frontiers P505-15 manufacturer in measurement and assessment. Edited by: Magurran AE, McGill BJ. Oxford: Oxford University Press; 2011:221–236. 2. Alexander E, Stock A, Breiner HW, Behnke A, Bunge J, Yakimov MM, Stoeck T: Microbial eukaryotes in the hypersaline anoxic L’Atalante deep-sea basin. Environ Microbiol 2009, 11:360–381.PubMedCrossRef 3. Edgcomb V, Orsi W, Leslin C, Epstein S, Bunge J, Jeon SO, Yakimov MM, Behnke A, Stoeck T: Protistan community patterns within the brine and halocline

of deep hypersaline anoxic basins in the eastern Mediterranean Sea. Extremophiles 2009, 13:151–167.PubMedCrossRef 4. Camerlenghi A: Anoxic basins of the eastern

Mediterranean: geological framework. Mar Chem 1990, 31:1–19.CrossRef Quisinostat molecular weight 5. La Cono V, Smedile F, Bortoluzzi G, Arcadi E, Maimone G, Messina E, Borghini M, Oliveri E, Mazzola S, L’Haridon S, et al.: Unveiling microbial life in new deep-sea hypersaline Lake Thetis. Part I: Prokaryotes and environmental settings. Environ Microbiol 2011,13(8):2250–2268.PubMedCrossRef 6. van der Wielen PW, Bolhuis H, Borin S, Daffonchio D, Corselli C, Giuliano L, D’Auria G, de Lange GJ, Huebner A, Varnavas SP, et al.: The enigma of prokaryotic life in deep hypersaline anoxic basins. Science 2005,307(5706):121–123.PubMedCrossRef 7. Azam F, Fenchel T, Field J, Gray J, Meyer-Reil L, Thingstad F: The ecological role of water column microbes in

the sea. Mar Ecol Prog Ser 1983, 10:257–263.CrossRef 8. Corliss JO: Biodiversity and biocomplexity of the protists and an overview of their significant roles in maintenance of our biosphere. Acta Protozool 2002,41(3):199–220. 9. Finlay BJ, Corliss JO, Esteban G, Fenchel T: Biodiversity at the microbial level: the number of free-living ciliates in the biosphere. Ouart Rev Biol 1996, 71:221–237.CrossRef 10. Lynn DH, Gilron GL: A brief review of approaches using ciliated protists to assess aquatic ecosystem health. J Aquatic Ecosyst Health 1992, 1:263–270.CrossRef 11. Doherty Cytoskeletal Signaling inhibitor M, Cosatas BA, McManus GB, Katz LA: Culture independent assessment of planktonic ciliate diversity in coastal northwest Atlantic waters. Aquat Microb Ecol 2007, 48:141–154.CrossRef 12. Fenchel T, Finlay BJ: The diversity of microbes: resurgence of the phenotype. Phil Trans Roy Soc Lond B Biol Sci 2006,361(1475):1965–1973.CrossRef 13. Finlay BJ: Global dispersal of free-living microbial eukaryote species. Science 2002,296(5570):1061–1063.PubMedCrossRef 14. Foissner W, Chao A, Katz LA: Diversity and geographic distribution of ciliates (Protista: Ciliophora). Biodiv Conserv 2008, 17:345–363.CrossRef 15.

PubMedCrossRef 53 Wunder C, Eichelbronner O, Roewer N: Are IL-6,

PubMedCrossRef 53. Wunder C, Eichelbronner O, Roewer N: Are IL-6, IL-10 and PCT plasma concentrations reliable for outcome prediction in severe sepsis? A comparison with APACHE III and SAPS II. Inflamm Res 2004, 53:158–163.PubMedCrossRef 54. Novotny A, Emanuel K, Matevossian E, Kriner M, Ulm K, Bartels

H, Holzmann B, Weighardt H, Siwert J-R: Use of procalcitonin for early prediction of lethal outcome of postoperative sepsis. Am J Surg 2007, 194:35–39.PubMedCrossRef Competing interests The authors declare that they have no competing interests. Authors’ contribution JS and KM are equally engaged into the study: Study design, data collection, statistical analysis, Selleckchem YH25448 data interpretation, manuscript preparation, literature search, and funds collection. Both authors read and approved the final manuscript.”
“Background Sigmoid volvulus in pregnancy is a rare but serious complication associated with a significant maternal PX-478 and fetal

mortality [1]. The fundamental problem of sigmoid volvulus in pregnancy until is that of delay in presentation and further delay in diagnosis leading to ischemia of the colon, which requires bowel resection and colostomy as seen in most of the reported cases [2–20]. Timely surgical intervention is essential to reduce maternal and fetal morbidity

and mortality [1]. Perforation, peritonitis and sepsis can be the maternal complications if intervention is not done early in the course of the disease. The fetal complications include preterm delivery, intrauterine death and neonatal sepsis. We have reviewed the available literature on this subject and report another case of a 30-week pregnant lady who presented to us with complicated sigmoid volvulus (Table 1). There is a need to increase the awareness amongst the general practitioners and community obstetricians for this potentially life threatening condition. A high index of suspicion and judicious use of modern radiological imaging may help make an early diagnosis and improve the maternal and fetal outcomes.

**Classification of cefazolin as ‘active’ or ‘less active’: When

**Classification of cefazolin as ‘active’ or ‘less active’: When difference in cleavage rates (fluorescence change) in the absence and presence of cefazolin was minimal, antibiotic predicted to be ‘active’. Drastically lowered cleavage rate in presence of cefazolin compared to when probe assayed alone led to prediction of cefazolin as ‘less active’ respectively (also see Figure 2). Details of Disk Diffusion results are presented in Table 3. Bacteria-free controls (PBS only) were included in each assay-set to account for non-specific probe cleavage that may occur. As expected, a negligible fluorescence change over time was observed. Comparison of cleavage rates (mRFU/min) for

#1, #2 and the PBS only control are shown in Additional file 1: Figure S1. Nitrocefin test for detection of β-lactamase validates results from β-LEAF Defactinib assay In order to validate the β-lactamase phenotypes determined by the β-LEAF assay, a CLSI recommended β-lactamase screening method, the chromogenic nitrocefin test, was utilized [41]. All bacterial isolates that were strongly positive by the β-LEAF assay were also found to be positive by nitrocefin conversion with the nitrocefin disks, showing a change in colour from yellow to deep orange in a positive reaction for β-lactamase (Table 1, right-most

column). Comparison of conventional disk diffusion and β-LEAF assay results In order to compare predictions of cefazolin activity by the β-LEAF assay to a conventional AST method, we performed cefazolin disk diffusion check details assays with the S. aureus isolates. Based on respective zone of inhibition diameters, each isolate was classified as susceptible, intermediate or resistant using the CLSI zone interpretive criteria (Table 3, Additional file 2: Figure S2). Interestingly, all the isolates

fell in the cefazolin ‘susceptible’ range with this methodology (Table 3). Table 3 Cefazolin disk diffusion results S. aureus isolate # Zone of inhibition diameter (mm) AS* Zone edge Interpretation as per zone edge test criteria& 1 21.5 ± 1.0 S Sharp β 2 31.0 ± 1.0 S Fuzzy   3 33.5 ± 0.5 S Fuzzy   4 33.0 ± 2.0 S Fuzzy   5 32.5 ± 0.5 S Fuzzy   6 36.5 ± 0.5 Mannose-binding protein-associated serine protease S Sharp β 7 32.0 ± 0.5 S Fuzzy   8 39.5 ± 1.5 S Fuzzy   9 29.5 ± 1.5 S Fuzzy   10 41.5 ± 0.5 S Fuzzy   11 34.5 ± 2.5 S Little fuzzy Weak β? 12 41.0 ± 1.6 S Fuzzy   13 32.5 ± 0.5 S Fuzzy   14 33.0 ± 0.0 S Fuzzy   15 35.5 ± 2.5 S Fuzzy   16 36.5 ± 0.5 S Fuzzy   17 36.5 ± 0.5 S Fuzzy   18 33.5 ± 0.5 S Sharp β 19 31.0 ± 0.0 S Sharp β 20 20.5 ± 0.3 S Sharp β 21 38.0 ± 1.0 S Fuzzy   22 34.0 ± 1.1 S Little fuzzy Weak β? 23 33.5 ± 1.5 S Fuzzy   24 34.5 ± 1.5 S Fuzzy   25 30.5 ± 0.5 S Fuzzy   26 34.0 ± 0.0 S Fuzzy   27 36.0 ± 2.0 S Little fuzzy/sharpish Weak β? *The Antibiotic Susceptibility (AS) was determined using the CLSI Zone Diameter Interpretive Criteria for Cefazolin Disk Diffusion [41].

Horm Res 2003, 60:174–180 PubMedCrossRef 28 Yoon SK, Lim NK, Ha

Horm Res 2003, 60:174–180.PubMedCrossRef 28. Yoon SK, Lim NK, Ha SA, Park YG, Choi JY, Chung KW: The human cervical cancer oncogene protein is a biomarker for human hepatocellular carcinoma. Cancer Res 2004, 64:5434–5441.PubMedCrossRef 29. Marrero JA, Romano PR, Nikolaeva O, Steel L, Mehta A, Fimmel CJ: Gp73, a resident golgi glycoprotein, is a novel serum marker for hepatocellular carcinoma. www.selleckchem.com/products/th-302.html J Hepatol 2005, 43:1007–1012.PubMedCrossRef 30. Yamagamim H, Moriyama M, Matsumura H, Aoki H, Shimizu T, Saito T: Serum concentrations of human hepatocyte growth factor is

a useful indicator for predicting the occurrence of hepatocellular carcinomas in c-viral chronic liver diseases. Cancer 2002, 95:824–834.PubMedCrossRef buy Staurosporine 31. Moriyama M, Matsumura H, Watanabe A, Nakamura H, Arakawa

Y, Oshiro S: Detection of serum and intrahepatic KL-6 in anti-HCV positive patients with hepatocellular carcinoma. Hepatol Res 2004, 30:24–33.PubMedCrossRef 32. Semela D, Dufour JF: Angiogenesis and hepatocellular carcinoma. J Hepatol 2004, 41:864–880.PubMedCrossRef 33. Hann HW, Lee J, Bussard A, Liu C, Jin YR, Guha K: Preneoplastic markers of hepatitis B virus-associated hepatocellular carcinoma. Cancer Res 2004, 64:7329–7335.PubMedCrossRef 34. Hu WQ, Peng CW, Li Y: The expression and significance of P-glycoprotein, lung resistance protein and multidrug resistance-associated protein in gastric cancer. J Exp Clin Cancer Res 2009, 28:144–150.PubMedCrossRef 35. Li W, Gomez E, Zhang Z: Immunohistochemical expression of stromal cell-derived factor-1 (SDF-1) and CXCR4 ligand receptor system in hepatocellular carcinoma. J Exp Clin Cancer Res 2007, 26:527–533.PubMed 36. Li N, Long Y, Fan X, Liu H, Li C, Chen L, Wang Z: Proteomic analysis of differentially expressed proteins in hepatitis B virus-related hepatocellular carcinoma tissues. J Exp Clin www.selleck.co.jp/products/Metformin-hydrochloride(Glucophage).html Cancer Res 2009, 28:122–132.PubMedCrossRef 37. Qiu FM, Yu JK, Chen YD, Jin QF, Sui MH, Huang J: Mining novel biomarkers for prognosis of gastric cancer with

serum proteomics. J Exp Clin Cancer Res 2009, 28:126–133.PubMedCrossRef 38. Rybakin V, Clemen CS: Coronin proteins as multifunctional regulators of the cytoskeleton and membrane trafficking. Bioessays 2005, 27:625–632.PubMedCrossRef 39. Spoerl Z, Stumpf M, Noegel AA, Hasse A: Oligomerization, F-actin interaction, and membrane association of the ubiquitous mammalian coronin 3 are mediated by its carboxyl terminus. J Biol Chem 2002, 277:48858–48867.PubMedCrossRef 40. Thal D, Xavier CP, Rosentreter A, Linder S, Friedrichs B, Waha A, Pietsch T, Stumpf M, Noegel A, Clemen C: Expression of coronin-3 (coronin-1C) in diffuse gliomas is related to malignancy. J Pathol 2008, 214:415–424.PubMedCrossRef Competing interests The authors declare that they have no competing interests.

neoformans for an additional 1 hr and subsequent microscopic imag

neoformans for an additional 1 hr and subsequent microscopic imaging. Collection of human peripheral blood monocytes and phagocytosis Monocytes were isolated

by Ficoll-Hypaque (GE Healthcare, Piscataway, NJ) density gradient centrifugation as described previously [30]. Briefly, diluted venous blood from one healthy donor was diluted with Hank’s balanced salt solution (Mediatech, Herndon, Va) and was layered on top of Ficoll-Hypaque (GE Healthcare) at a 1:1 ratio and centrifuged at 2000 rpm/4°C for 15 minutes without brake. The monocyte layer was removed and red blood cells were lysed using lysing buffer (0.155 M NH4Cl pH 7.4). Cells were washed three times with Hank’s balanced salt solution and suspended in RPMI (Mediatech) media supplemented with 10% fetal calf serum (Gemini Bioproducts, West Sacramento, Ca) and cells were then plated on poly-lysine coverslip-bottom PS-341 in vivo MaTtek plates (Ashland, MA)

at a density of 2 × 105 per well in feeding media and allowed to adhere at 37°C and 10% CO2 for 6 days prior to incubation with C. neoformans, using 18B7 (10 ug/ml) or 20% human serum, for 1 hr and subsequent microscopic imaging. This study was done with the approval of our institutional review board committee at the Albert Einstein College of Medicine and prior consent was obtained from blood donors. Time-lapse imaging For live cell imaging, phagocytosis assays were done as described [9]. Briefly, 105 HPBM were plated on polylysine FG-4592 coated coverslip bottom MatTek plates and allowed to adhere for 6 days. The media was then removed and replaced with fresh media containing C. neoformans cells (C. Aldol condensation neoformans to HPBM ratio of 10:1) along with monoclonal antibody (mAb) against the cryptococcal capsule (mAb 18B7, 50 μg/ml). C. neoformans

were opsonized with either mAb 18B7 or 20% guinea pig serum as indicated above. HPBMs and C. neoformans were then incubated together for 30 min at 4°C to synchronize phagocytosis, followed by 60 min incubation at 37°C to allow for completion of phagocytosis. This was followed by two washes with fresh media (1 ml each), and replenishment with 2 ml feeding media. The plates were then taken for time-lapse imaging every 4 minutes using an Axiovert 200 M inverted microscope and photographed with an AxiocamMR camera controlled by the Axio Vision 4.4 software (Carl Zeiss Micro Imaging, NY). This microscope was housed in a Plexiglas box and the temperature was stabilized at 37°C with a forced air heater system. The plate lid was kept in place to prevent evaporation, and 5% CO2 was delivered to a chamber locally at the culture dish. Quantitative analysis of phagosomal extrusion and cell to cell spread was carried out by compiling all the movies and counting the number of macrophages with internalized C.

The main reason for cancellation was surgeon’s unavailability

The main reason for cancellation was surgeon’s unavailability

[28]. Changing the operating theatre policy, as demonstrated in this article, allows surgeons to designate and inform the patient more accurately the time of his/her operation. However, it did not necessarily reduce the waiting times to surgery. We feel that provision of a second emergency theatre at all times would be an effective solution to this problem. Patients would be operated upon promptly. This would reduce waiting Talazoparib research buy times to surgery and facilitate quicker discharges from hospital, thereby increasing turnover. This would also be satisfactory for the patients; bed management for the elective patients, thereby increasing volumes of elective work load and shortening waiting list times. The increased costs involved in running the second additional theatres should be balanced against the cost of reduced length of hospital stay. Taking an example from emergency laparoscopic cholecystectomy versus elective cholecystectomy after conservative management, the increased immediate operative cost is neutralized by the reduced length of stay and quicker return to work [29]. More detailed cost – benefit analysis involving multiple hospitals and larger number of patients would be required to lend creditable evidence to support this belief. Acknowledgements We thank all

the medical and nursing staff of the wards and theatres of the surgical many services for taking care of patients and helping in data collection. We thank Mr Ajit Abraham & Mr Mike Walsh, Consultant Surgeons TGF-beta cancer for spearheading the theatre change programme and Ms Ceri Cranston, Theatre Manager for implementing the changes with rigor. References 1. Wyatt MG, Houghton PW, Brodribb AJ: Theatre delay for emergency general surgical patients: a cause for concern? Ann R Coll Surg Engl 1990,72(4):236–8.PubMed 2. American College of Surgeons Trauma Program [http://​www.​facs.​org/​trauma] 3. Bhattacharyya T, et al.:

The value of the dedicated orthopaedic trauma operating room. J Trauma 2006,60(6):1336–40. discussion 1340–1CrossRefPubMed 4. The Report of the National Confidential Enquiry into Perioperative Deaths 1990 NCEPOD, London; 1992. 5. Sweetnam DI, Williams JR, Britton DC: An audit of the effect of a 24-hour emergency operating theatre in a district general hospital. Ann R Coll Surg Engl 1994,76(2 Suppl):56–8.PubMed 6. Lovett BE, Katchburian MV: Emergency surgery: half a day does make a difference. Ann R Coll Surg Engl 1999,81(1):62–4.PubMed 7. Calder FR, Jadhav V, Hale JE: The effect of a dedicated emergency theatre facility on emergency operating patterns. J R Coll Surg Edinb 1998,43(1):17–9.PubMed 8. Barlow AP, et al.: An emergency daytime theatre list: utilisation and impact on clinical practice. Ann R Coll Surg Engl 1993,75(6):441–4.PubMed 9. Scriven MW, et al.

crescentus NA1000 were used The figure was generated using the W

crescentus NA1000 were used. The figure was generated using the WebLogo server [42], and the height of the residue symbol indicates the degree of conservation within the ortologous groups. The

sequence numbering shown below the alignment corresponds to the respective C. crescentus NA1000 proteins. The complete representation of the motifs for the CzrA and NczA orthologous groups are shown in Additional file 2: Figure S1. (C) Cartoon representation of the CzrA structure model in which the conserved motifs MI-MV and the Loop are colored in yellow. The sub-domains DC, DN, PC1, PC2, PN1 and PN2 are Anlotinib colored in yellow, blue, dark green, red, violet and orange, respectively. The CzrA structure model was obtained using the Phyre2 program with CusA structure as a model (PDB: 3 k07, [25]). The structure was generated using PyMOL [43]. The secondary structure elements indicated were predicted using the PHYRE2

program [44]; red ovals and amino acid sequences indicate α-helix; orange arrows and amino acid sequences indicate β-strands. In order to localize the identified signatures in the CzrA protein structure, we performed a homology MLN2238 order modeling analysis utilizing the structure of E. coli CusA as model (PDB: 3 k07), since it is the only metal-transporting RND protein structure so far available in the data bases. All of the motifs described above, with the exception of MV, are located in the periplasmic domain of CzrA structural model (Figure 6C). MV is located in TM8 in CzrA (Figure 6C), which in E. coli CusA suffers a significant conformational change when it binds Cu+ or Ag+, and was proposed to be involved in transmembrane signaling and in initiation of proton translocation across the membrane

[25]. MI and MII are located in two close loops in the sub-domain PN1, MIII is located in the sub-domain DN and MIV is located in the sub-domain-PC2 (Figure 6C). The Etofibrate PC2 sub-domain in E. coli CusA was proposed to move, creating a cleft between PC1 and PC2 when CusA binds to Cu+ or Ag+[25]. The most conspicuous difference between the CzrA and NczA groups is the length of the loop located in PN2, called here Large Loop for CzrA and Small loop for NczA. The periplasmic PN2 region is involved in the interaction between E. coli CusA and one molecule of the CusB dimer [25, 45]. When we superimpose the CzrA model on the CusAB2 complex structure (PDBID: 3NE5), the results suggest that the Large Loop could affect the interaction between CzrA and the adaptor protein (not shown). The predicted adaptors for the C. crescentus HME-RND systems, CzrB and NczB, share no significant amino acid sequence identity with CusB [45]. Nevertheless, most of the interface residues at the sub-domain DC in CusA involved in the interaction with one molecule of the CusB dimer are conserved in the CzrA and NczA orthologs, although the two residues located in PN2, D155 and R147, are not conserved in members of either group.

It has recently been proposed as the official primary barcoding m

It has recently been proposed as the official primary barcoding marker for fungi (Deliberation of 37 mycologists from 12 countries at the Smithsonian’s Conservation and Research Centre, Front Royal, Virginia, May 2007). More than 100 000 fungal ITS sequences generated by conventional Sanger sequencing are deposited in the International Nucleotide Sequence Databases and/or

other databases [11], providing a large reference material for identification of fungal taxa. However, these data are to some extent hampered by misidentifications or technical errors such as mixing of DNA templates or sequencing errors [12]. Furthermore, a large amount of partial ITS sequences generated by next-generation sequencing has Selleck MI-503 recently been deposited in public sequence databases. The ITS region includes the ITS1 and ITS2 regions, separated by the 5.8S gene, and is situated between the 18S (SSU) and 28S (LSU) genes in the nrDNA repeat unit (Figure 1). The large number of ITS copies per cell (up to 250; [13]) makes the region an appealing target for sequencing environmental substrates where the quantity of DNA

present is low. The entire ITS region has commonly been targeted with traditional Sanger sequencing approaches and typically ranges between 450 and 700 bp. Either the ITS1 or the ITS2 region have been targeted in recent high-throughput sequencing check details studies [14–17], because the entire ITS region is still too long for 454 sequencing or other high-throughput sequencing methods. Using high-throughput sequencing, thousands of sequences can be analysed from a single environmental sample, enabling in-depth analysis of the fungal diversity. Various primers Farnesyltransferase are used for amplifying the entire or parts of the ITS region (Figure 1). The most commonly used primers were published

early in the 1990′s (e.g. [18, 19] when only a small fraction of the molecular variation in the nrDNA repeat across the fungal kingdom was known. Several other ITS primers have been published more recently [20] but have not been used extensively compared to the earlier published primers. However, little is actually known about the potential biases that commonly used ITS primers introduce during PCR amplification. Especially during high-throughput sequencing, where quantification (or semi-quantification) of species abundances is also possible to a certain degree (although hampered by factors like copy-number variation), primer mismatches might potentially introduce large biases in the results because some taxonomic groups are favoured during PCR. Our main focus in this study is on the two dominating taxonomic groups of fungi in the Dikarya, Ascomycota and Basidiomycota.

FEMS Microbiol Lett 1996, 141:151–156 PubMedCrossRef 22 Lund T,

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