Furthermore, SA1101 showed an inhibitory effect toward the growth

Furthermore, SA1101 showed an inhibitory effect toward the growth of osteoblastic cells and had greater properties of adhesion to those cells as compared to ATCC49456. Conclusions.  These Small molecule library results suggest that S. mitis SA1101 is a possible etiological agent and caused osteomyelitis in the present case. “
“International Journal of Paediatric Dentistry 2012; 22: 318–323 Background.  Mucocele is a common oral lesion in children and adolescents. Different techniques have been described for the treatment; however, all of them are invasive. Aim.  This work studied the efficacy of micro-marsupialization

for the treatment for mucoceles in paediatric patients. Design.  A retrospective review was performed using the clinical records of patients aged between 0 and 18 years with a clinical diagnosis of mucocele. The following data were obtained: age, gender, location and size of the lesion, duration of mucocele development, and type of treatment and its results. Results.  The mean age of the patients was 11.1 ± 3.95 years. Mucoceles were found in the lower lip (83.7%), buccal mucosa (11.6%), and tongue (4.7%). From the overall cohort of 86 cases, 33 were treated by micro-marsupialization,

of which five developed a recurrence that required surgical excision. The other 53 cases were treated by surgical excision, and three of these had recurrent disease. No statistically significant difference was found between the

treatment methods. Conclusions.  Micro-marsupialization can be 17-DMAG (Alvespimycin) HCl used to treat mucoceles in paediatric dentistry. It is simpler to perform, minimally invasive, Saracatinib in vitro requires no local infiltration of anaesthesia, has a lower postoperative complications rate, and is well-tolerated by patients. “
“Current molar hypomineralisation (MH) indices do not guide clinicians in management of affected dentitions, and treatment is based on individual judgment. The aims of this study were to describe characteristics of MH and molar incisor hypomineralisation (MIH) and trial the new Molar Hypomineralisation Severity Index (MHSI). First permanent molars (FPMs) and permanent incisors (PIs) in 283 affected children were examined for hypomineralisation characteristics [defect colour, location, post-eruptive breakdown (PEB); restorations placed/replaced/atypical; sensitivity]. The MHSI scores were compared with treatment received (152 children). Mean (SD) affected teeth/dentition were as follows: FPMs: 3.2 (1.0) and PIs: 1.6 (1.6). Affected FPMs showed no arch or quadrant predilection; maxillary central PIs were affected particularly. As affected FPMs/dentition increased, MIH diagnoses also increased (P = 0.009). Among FPMs, defects most prevalent were brown (47%) and cuspal (74%); 67% showed PEB. Before study entry, 43% of FPMs had restorations placed/replaced. Among PIs, white defects were common (65%) on smooth surfaces; sensitivity was rare.

Quantitative analysis was performed

Quantitative analysis was performed LDK378 molecular weight using the GeneAmp®7000 Sequence Detection System (PE Applied Biosystems) with PCR conditions of 95 °C for 15 s and 60 °C for 1 min for 40 cycles. Three independent experiments were carried out. Each sample was examined in triplicate, using relative quantification analysis. The plasmid pSilent1 (Nakayashiki et al., 2005) was obtained from the Fungal Genetics Stock Center (McCluskey, 2003). A 340-bp fragment

from the Tas-acdS encoding region was cloned into pSilent1 in sense and reverse/complementary orientations on both sides (XhoI/HindIII sites and StuI/ApaI sites) of the 147-bp intron 2 of the cutinase gene from Microdochium oryzae driven by the PtrpC promoter. The coding region, in the sense orientation, was amplified by PCR with the primers ACCXhoI (5′-CCGCTCGAGCACAAGCCCACGCTGGCAAACC-3′) and ACCHindIII (5′-CCAAGCTTTGGCAGCAGTGAATTTAGC-3′). The coding region in the

antisense orientation was amplified by PCR with the primers ACCApaI (5′-AAAGGGCCCCACAAGCCCACGCTGGCAAACC-3′) and ACCStuI (5′-AAGGCCTTGGCAGCAGTGAATTTAGC-3′). Microprojectile bombardment of intact T. asperellum T203 conidia with the pSilent1-Tas-acdS/RNAi plasmid was performed as described in Viterbo et al.(2002). Silencing of Tas-acdS in ACC-induced cultures was analyzed by comparing the relative gene expression of Tas-acdS/RNAi XL765 order lines to the wild type by real-time RT-PCR using the same primer sets as described above. Intron-free cDNA was obtained from total RNA extracted from T. asperellum cultures grown in the presence of ACC (3 mM) as the sole nitrogen source. The coding region was amplified by PCR (5′-ATGGCTACCCTCAACATCC-3′, 5′-TCAGTCTAAAAGAGAGGAATACGC-3′), Unoprostone subcloned in pGEM-T Easy vector (Promega) and cloned in the pALTER-EX1 (Promega) vector in NdeI/NcoI sites under the control of the tac promoter. The hybrid plasmid was then transformed into JM109 cells and ACCD activity

was tested as described in the next section. For ACCD activity determination in recombinant E. coli and Pseudomonas putida UW4, bacteria were grown as described in Penrose & Glick (2003). For determination of ACCD activity in Trichoderma, a 20-μL spore suspension was inoculated in 10-mL synthetic medium (SM; Yedidia et al., 1999) and the culture was grown for 48 h. The washed mycelia were then transferred to 5 mL of SM without ammonium and with 0.3–3 mM ACC. At the end of the induction period, the cultures were resuspended in half volume of Tris buffer 0.1 M (pH 8.5) and homogenized using an ULTRA-TURRAX apparatus (Janke & Kunkel, Staufen, Germany). Toluene (25 μL) was added to a 200-μL aliquot and vortexed vigorously for 30 s. ACC (20 μL of 0.5 M solution) was added, and after an incubation period of 15 min at 30 °C, 1 mL of 0.56 N HCl was added. The lysates were centrifuged (10 000 g, 10 min) and 1 mL of the supernatant was mixed with 800 μL of 0.

, 2011) The phylogenetic composition of the extracted DNA in rel

, 2011). The phylogenetic composition of the extracted DNA in relation

to the original bacterial community has however received less attention. One major problem in assessing this is the fact that it is very difficult to know the true community composition. In two recent studies, it was shown that a mechanical bead-beating step during cell lysis resulted in increased complexity of extracted DNA as evidenced by an increased number of distinct bands in PCR-DGGE profiles (Ariefdjohan et al., 2010; Smith et al., 2011). The fact that different extraction procedures performed on the same fecal sample may lead to different estimations of the bacterial community composition is not surprising, but may well be disturbing for comparisons between separate studies. Within a study, it is most probable that the same DNA Selleck CHIR-99021 extraction method be used throughout; however, other parameters that may affect extraction, such as storage conditions of fecal samples, may vary. It is for instance common practice, mainly for practical reasons, to freeze fecal samples immediately after sampling and then collectively extract the DNA and perform downstream analysis such as sequencing or qPCR, at some later stage (Mariat et al., 2009; Santacruz et al., 2009). In this study, the effect of freezing fecal samples prior to DNA extraction was evaluated for alterations in DNA

recovery and bacterial community composition as determined by downstream quantitative PCR analysis. Fecal samples were obtained from three healthy adult volunteers (two women, one man), homogenized thoroughly in four volumes diluent (0.85% NaCl, 0.1% peptone), see more centrifuged at 300 g for 2 min to remove large debris, and finally 0.5 mL of aliquots (average 8 mg dry weight) were pelleted at 10 000 g for 5 min (Fig. 1). Extraction of DNA was performed immediately with three different extraction methods (five replicates), or samples were frozen at −20 °C for 53 ± 5 days (F) prior to extraction.

Methods used for DNA extraction were M: PowerSoil® DNA Isolation kit (MO BIO Laboratories, Carlsbad), Q: QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany), and B: Modified QIAamp DNA Stool Mini Kit extraction procedure with the incorporation of a bead-beating step to potentially improve Methisazone cell lysis (Leser et al., 2000). Briefly, bead-beating was performed by adding 500 μL autoclaved 0.1 mm zirconia silica beads (Biospec Products Inc., Bartlesville, OK) and 30 μL of 10% sodium dodecyl sulfate and processing for 4 min. at 30 cycles per second on a Mixer Mill MM 300 (Retsch GmbH, Haan, Germany). Extractions were performed as directed by the suppliers with minor modifications, including standardized initial sample size and elution in 200 μL, 10 mM Tris, to allow better comparison of the methods. For extractions with method M, bacterial cells were treated in a Mixer Mill MM 300 (4 min at 30 cycles per second) and not the suggested Vortex adaptor.

, 2011) The phylogenetic composition of the extracted DNA in rel

, 2011). The phylogenetic composition of the extracted DNA in relation

to the original bacterial community has however received less attention. One major problem in assessing this is the fact that it is very difficult to know the true community composition. In two recent studies, it was shown that a mechanical bead-beating step during cell lysis resulted in increased complexity of extracted DNA as evidenced by an increased number of distinct bands in PCR-DGGE profiles (Ariefdjohan et al., 2010; Smith et al., 2011). The fact that different extraction procedures performed on the same fecal sample may lead to different estimations of the bacterial community composition is not surprising, but may well be disturbing for comparisons between separate studies. Within a study, it is most probable that the same DNA find more extraction method be used throughout; however, other parameters that may affect extraction, such as storage conditions of fecal samples, may vary. It is for instance common practice, mainly for practical reasons, to freeze fecal samples immediately after sampling and then collectively extract the DNA and perform downstream analysis such as sequencing or qPCR, at some later stage (Mariat et al., 2009; Santacruz et al., 2009). In this study, the effect of freezing fecal samples prior to DNA extraction was evaluated for alterations in DNA

recovery and bacterial community composition as determined by downstream quantitative PCR analysis. Fecal samples were obtained from three healthy adult volunteers (two women, one man), homogenized thoroughly in four volumes diluent (0.85% NaCl, 0.1% peptone), buy 3-Methyladenine centrifuged at 300 g for 2 min to remove large debris, and finally 0.5 mL of aliquots (average 8 mg dry weight) were pelleted at 10 000 g for 5 min (Fig. 1). Extraction of DNA was performed immediately with three different extraction methods (five replicates), or samples were frozen at −20 °C for 53 ± 5 days (F) prior to extraction.

Methods used for DNA extraction were M: PowerSoil® DNA Isolation kit (MO BIO Laboratories, Carlsbad), Q: QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany), and B: Modified QIAamp DNA Stool Mini Kit extraction procedure with the incorporation of a bead-beating step to potentially improve Abiraterone datasheet cell lysis (Leser et al., 2000). Briefly, bead-beating was performed by adding 500 μL autoclaved 0.1 mm zirconia silica beads (Biospec Products Inc., Bartlesville, OK) and 30 μL of 10% sodium dodecyl sulfate and processing for 4 min. at 30 cycles per second on a Mixer Mill MM 300 (Retsch GmbH, Haan, Germany). Extractions were performed as directed by the suppliers with minor modifications, including standardized initial sample size and elution in 200 μL, 10 mM Tris, to allow better comparison of the methods. For extractions with method M, bacterial cells were treated in a Mixer Mill MM 300 (4 min at 30 cycles per second) and not the suggested Vortex adaptor.

, 2011) The phylogenetic composition of the extracted DNA in rel

, 2011). The phylogenetic composition of the extracted DNA in relation

to the original bacterial community has however received less attention. One major problem in assessing this is the fact that it is very difficult to know the true community composition. In two recent studies, it was shown that a mechanical bead-beating step during cell lysis resulted in increased complexity of extracted DNA as evidenced by an increased number of distinct bands in PCR-DGGE profiles (Ariefdjohan et al., 2010; Smith et al., 2011). The fact that different extraction procedures performed on the same fecal sample may lead to different estimations of the bacterial community composition is not surprising, but may well be disturbing for comparisons between separate studies. Within a study, it is most probable that the same DNA see more extraction method be used throughout; however, other parameters that may affect extraction, such as storage conditions of fecal samples, may vary. It is for instance common practice, mainly for practical reasons, to freeze fecal samples immediately after sampling and then collectively extract the DNA and perform downstream analysis such as sequencing or qPCR, at some later stage (Mariat et al., 2009; Santacruz et al., 2009). In this study, the effect of freezing fecal samples prior to DNA extraction was evaluated for alterations in DNA

recovery and bacterial community composition as determined by downstream quantitative PCR analysis. Fecal samples were obtained from three healthy adult volunteers (two women, one man), homogenized thoroughly in four volumes diluent (0.85% NaCl, 0.1% peptone), selleckchem centrifuged at 300 g for 2 min to remove large debris, and finally 0.5 mL of aliquots (average 8 mg dry weight) were pelleted at 10 000 g for 5 min (Fig. 1). Extraction of DNA was performed immediately with three different extraction methods (five replicates), or samples were frozen at −20 °C for 53 ± 5 days (F) prior to extraction.

Methods used for DNA extraction were M: PowerSoil® DNA Isolation kit (MO BIO Laboratories, Carlsbad), Q: QIAamp DNA Stool Mini Kit (QIAGEN, Hilden, Germany), and B: Modified QIAamp DNA Stool Mini Kit extraction procedure with the incorporation of a bead-beating step to potentially improve Atazanavir cell lysis (Leser et al., 2000). Briefly, bead-beating was performed by adding 500 μL autoclaved 0.1 mm zirconia silica beads (Biospec Products Inc., Bartlesville, OK) and 30 μL of 10% sodium dodecyl sulfate and processing for 4 min. at 30 cycles per second on a Mixer Mill MM 300 (Retsch GmbH, Haan, Germany). Extractions were performed as directed by the suppliers with minor modifications, including standardized initial sample size and elution in 200 μL, 10 mM Tris, to allow better comparison of the methods. For extractions with method M, bacterial cells were treated in a Mixer Mill MM 300 (4 min at 30 cycles per second) and not the suggested Vortex adaptor.

Urgent referral would allow confirmation of a diagnosis of HIV in

Urgent referral would allow confirmation of a diagnosis of HIV in an infant and treatment to prevent disease progression. Individual feedback was sent to the units who sent guidelines, TGF-beta inhibitor to allow them to improve their guidelines. Two units asked for a template to produce local guidelines. In summary, mother-to-child transmission of HIV is preventable. All maternity units should have

local guidelines, based on the BHIVA/CHIVA pregnancy guidelines, to allow them to manage infants born to HIV-infected women. Other regions should review local guidelines to ensure that they give enough information to manage both low-risk and high-risk infants, together with information on how and when to seek expert advice. “
“Quantification of obligate biotrophic parasites has been a long-standing problem in plant pathology. Many attempts have been made to determine how much of a pathogen is present in infected plant tissue. Methods of quantification

selleck screening library included scoring disease symptoms, microscopic evaluation, determination of specific compounds like Ergosterol, and lately nucleic acid-based technologies. All of these methods have their drawbacks, and even real-time PCR may not be quantitative if for example the organism of interest has specific and differing numbers of nuclei in different infection structures. We applied reverse transcription (RT) real-time PCR to quantify Uromyces fabae within its host plant Vicia faba. We used three different genes, which have been shown to be constitutively expressed. Our analyses show an exponential increase of fungal material between 4 and 9 days post inoculation and thereafter reaching a steady state of around 45% of total RNA. We also used haustorium-specific genes to determine the amount

of haustoria present at each time point. These analyses parallel the development of the whole fungus with the exception of the steady-state level, which is only around 5% of the total RNA. This indicates that RT real-time PCR is a suitable method for quantification of obligate biotrophic parasites, and also for the differentiation of developmental stages. All higher organisms exhibit a more or less pronounced association with a plethora of symbiotic microorganisms, some of them beneficial, some of them neutral, and some of them pathogenic. While the determination Phenylethanolamine N-methyltransferase of the number of mutualistic or neutral symbionts has more of an academic value, accurate quantification of pathogen abundance is a critical issue in medicine and plant pathology. There have been numerous approaches to quantify the number of pathogens present in various host–parasite interactions at any given time point of pathogenesis. Traditionally, visual inspection and scoring of disease symptoms have been used to determine disease severity (Pei et al., 2002; Bock et al., 2008). Lately, this type of rating has been complemented by digital image analysis (Bock et al., 2008).

, Tokyo, Japan) with the significance criteria of the program (P<

, Tokyo, Japan) with the significance criteria of the program (P<0.05). Real-time PCR was performed using a 7900HT Fast real-time PCR system CAL-101 datasheet (Applied Biosystems). Reactions containing cDNAs from 100 ng total RNA and gene-specific primers were prepared with SYBR Green Realtime PCR Master Mix (Toyobo) according to the manufacturer’s protocol. The primers used are listed in Table S1. The thermal cycle settings used were as follows: initial denaturation at 95 °C for 1 min followed by 40 cycles of denaturation at 95 °C for 15 s, annealing at 56 °C for 15 s, and extension at 72 °C for

1 min. The expression of target genes was normalized to the endogenous 16S rRNA gene in each strain. The relative quantification of target gene expression was performed using the comparative cycle threshold (CT) method (Livak & Schmittgen, Wnt inhibitor 2001). blast searches revealed that the TF0022 ORF encodes a HTCS protein that shares homology with GppX from P. gingivalis. We sequenced a fragment containing TF0022 and the upstream

flanking region from the ATCC 43037 genome and compared the data with the existing database sequence (http://www.oralgen.lanl.gov). Although some minor differences were found at the nucleotide level, none altered any functional domains or conserved motifs in the encoding protein (Fig. 1a, DDBJ/GenBank ID: AB587729). Alignment of the TF0022 and GppX polypeptides Phospholipase D1 revealed a notable structural difference: the TF0022 protein lacks the N-terminal portion containing a transmembrane region and part of a putative periplasmic/sensor domain with a TPR motif (Fig. 1b). However, the immediate upstream ORF, TF0023, is predicted to encode a small polypeptide containing an N-terminal transmembrane region and a C-terminal TPR motif of almost the exact length needed to complement the ‘lost’ N-terminus of the TF0022 polypeptide. Indeed, both TF0022 and TF0023

were found to share homology with GppX, yielding similar blast scores (Fig. 1b). A single TPR motif typically consists of 34 amino acids (Das et al., 1998), and GppX from P. gingivalis harbors three tandem repeats of TPR (ranging from residues 155 to 254) at the center of the putative periplasmic domain (Fig. 1b). Interestingly, the TF0023 and TF0022 genes are in the same reading frame, and translation of the nucleotide sequences across the two ORFs uncovered an additional TPR motif when an 18-bp intergenic region was included (Fig. 1a). In P. gingivalis, one of the characteristic phenotypes of the disrupted gppX locus is enhanced autoaggregation (K. Nishikawa, unpublished data). In T. forsythia, ATCC 43037 wild-type cells gradually autoaggregate in broth cultures and eventually precipitate to the bottom of the test tubes. However, we noticed that the broth cultures of TF0022-ko mutant tended to precipitate faster than those of the wild-type strain.

healthtalkonlineorg) part of a new series of narrative on experi

healthtalkonline.org) part of a new series of narrative on experiences of using medicines and aimed to examine people’s experience of taking antidepressants. This paper focuses on treatment initiation. 38 people

with experience of LY2606368 taking antidepressants were interviewed. The study was approved by the UK Multi Centre Research Ethics Committee. A UK wide maximum variation sample was sought. The sample was obtained via doctors, support groups, social media and newsletters. Interviews were audio or video recorded, transcribed and returned to the participant for review. Emerging themes were identified using a ‘modified grounded theory’ approach and checked by each researcher and by members of the advisory panel. It took time before people began to feel AZD0530 supplier any benefits and they commonly experienced side effects. Sometimes people needed to try several different antidepressants before they found one that worked. It was important to have realistic

ideas for the first few weeks. Andrew’s doctor had pre-warned him that ‘you may just find that you’re fine but it may make you feel a little bit odd at first’ so he had an idea about what to expect. Talking to the doctor helped Stephen to keep in mind that it could take a while to notice any improvements in mood ‘I knew that if I took a tablet that day I wasn’t going to feel better tomorrow… it would take several weeks before it started to have any effect’. Some people noticed immediate benefit, and experience few, if any side effects. Sometimes being proactive and starting to ‘tackle the problem’ was enough to help people feel more positive. Several people noticed a gradual ‘lifting’ of their mood which could be ‘hard to pinpoint’. Roisin had tried a number of antidepressants that didn’t seem to make a difference, but when she began taking one that did suit her said she began to feel ‘almost normal’ after a few weeks. Lou described how her depression subsided after a few weeks of taking a new antidepressant, but overall

she said the medicine made her feel numb and distant. Overall, although there were benefits, many Diflunisal people were left feeling detached. Some people said they took time off from work to help them cope with their initial reaction to an antidepressant. Several people had found that varying the time of day when they took the antidepressant could help with the sleep related problems, or make other side effects such as nausea more bearable. Some people found that initial side effects continued, or the antidepressant didn’t seem to have a beneficial effect even after several weeks or months. The sample was chosen to represent a broad and diverse range of experiences, rather than to be numerically representative. Although people need a lot of support when starting antidepressants, none of the interviewees mentioned that they had received any support from a pharmacist during treatment initiation.

Of the 62 Twitter users, 50 (81%) health care professionals stopp

Of the 62 Twitter users, 50 (81%) health care professionals stopped using Twitter within six months of completing the module, although Twitter activity continued with 12 (19%) health care professionals, many of whom used it for both academic and social purposes. Among the topics covered in YouTube videos were: several different aspects of diabetes and macrovascular complications; a ‘one-to-one’

discussion on hypertension and cardiovascular disease; a ‘to camera’ piece on the links between diabetes and erectile selleck dysfunction; and, from an overseas student, a thought-provoking video on the burden of diabetes in South Africa, contrasting the levels of care available in the private and public sectors. The most popular YouTube video was entitled ‘Vascular

assessment of the lower limb and clinical diagnostics’ which had been viewed 1274 times by Bortezomib concentration August 2012. Of those who elected to create a Twitter account, the most active user had tweeted 257 times with 74 followers and following 86 other accounts. The least active Twitter user only tweeted six times but had secured 28 followers and was following 81 Twitter users. Data for 2010 and 2011 students are shown in Figure 1. Although there was a higher number of tweets posted by students in 2011 compared with students in 2010, the number of accounts that they followed, and the number of followers they attracted, many were broadly similar. In total, 13 (15%) health care professionals responded to an online questionnaire, four having selected YouTube and nine, Twitter (Figure 2). Eight students reported apprehension before embarking on the task but all expressed a sense of achievement and confidence in use of social media upon completion. Participants agreed that the assignment had changed their perception of social media, and that they could visualise

how it would be useful to them in their own practice, although one student expressed concern that using social media to communicate with patients could lead to urgent medical information not being acted upon within an appropriate timeframe. The exponential growth in internet use and, specifically, the rise in the use of social media including Twitter, Facebook, YouTube and similar channels that enable users to generate their own content and share with a vast audience have prompted many health care professionals to utilise this media for education4 as well as patient communication.9 As the intent of our postgraduate qualification is to enhance clinical expertise and improve patient care, we elected to incorporate social media within a postgraduate diabetes diploma and endeavour to assess its success.

In conclusion, our findings support the hypothesis of the acquisi

In conclusion, our findings support the hypothesis of the acquisition of genomic find more regions from other

pathogenic bacteria (E. coli or others) by horizontal transfers and reflect the genomic plasticity of EHEC or even E. coli strains. This variation in the genome contents of E. coli, suggested as a evolutionary strategy to better survive by Mokady et al. (2005), could lead to serious problems in public health and to the emergence of highly virulent new strains if one strain could acquire several strong virulence systems from different pathogenic bacteria, as it was dramatically illustrated by the 2011 Shiga toxin–producing E. coli O104:H4 German outbreak (Denamur, 2011; Rasko et al., 2011). During this study, Marjorie Bardiau was a PhD fellow of the ‘Fonds pour

la formation à la Recherche dans l’Industrie et dans l’Agriculture’ (FRIA). This study was funded by the Federal http://www.selleckchem.com/products/mitomycin-c.html Public Service of Health, Food Chain Safety and Environment (contract RF 6172), the European Network of Excellence EADGENE (European Animal Disease Genomics Network of Excellence for Animal Health and Food Safety) for the sequencing, and a grant ‘Crédits aux chercheurs’ FNRS (Fonds de Recherche Scientifique) 2008, no. 1363. “
“In silico analyses of several laccase promoter sequences have shown the presence of many different responsive elements differentially distributed along the promoter sequences. Analysis of Pleurotus ostreatus laccase promoter poxa1b extending around 1400-bp upstream of the start codon showed the presence of several putative response elements, such as 10 metal-responsive elements. Development of a system for in vivo analysis of P. ostreatus laccase promoter poxa1b by enhanced green fluorescent protein expression all was carried out, based on a polyethylene glycol–mediated procedure for fungal transformation.

Quantitative measurement of fluorescence expressed in P. ostreatus transformants grown in the presence and in the absence of copper sulfate was performed, demonstrating an increase in expression level induced by the metal. Twelve putative laccase genes have been identified in the recently sequenced Pleurotus ostreatus genome (http://www.jgi.doe.gov/sequencing/why/50009.html), one of which is annotated as a ferroxidase-like. The promoter regions of all the 11 P. ostreatus laccase genes, extending 500-bp upstream of the start codon, have been analyzed, revealing the presence of several putative response elements, differentially distributed along the promoter sequences (Piscitelli et al., 2011). All the analyzed P. ostreatus laccase promoters contain putative metal-responsive elements (MREs) with sequence homology to those reported in ascomycetous yeast.