Interestingly, the pRF size of non-deafferented V1 voxels increased slightly (~20% on average), although this effect appears weaker than that in previous single-unit recording reports. Area V2 also showed limited reorganisation. Remarkably, area V5/MT of the MD animal showed extensive activation compared

Trametinib manufacturer to controls stimulated over the part of the visual field that was spared in the MD animal. Furthermore, population receptive field size distributions differed markedly in area V5/MT of the MD animal. Taken together, these results suggest that V5/MT has a higher potential for reorganisation after MD than earlier visual cortex. “
“The current study examined the effects of pheromonal exposure on adult neurogenesis and revealed the Nutlin 3a role of the olfactory pathways on adult neurogenesis and behavior in the socially monogamous prairie vole (Microtus ochrogaster). Subjects were injected with a cell proliferation marker [5-bromo-2′-deoxyuridine (BrdU)]

and then exposed to their own soiled bedding or bedding soiled by a same- or opposite-sex conspecific. Exposure to opposite-sex bedding increased BrdU labeling in the amygdala (AMY), but not the dentate gyrus (DG), of female, but not male, voles, indicating a sex-, stimulus-, and brain region-specific effect. The removal of the main olfactory bulbs or lesioning of the vomeronasal organ (VNOX) in females reduced BrdU labeling in the AMY and DG, and inhibited the Tangeritin male bedding-induced BrdU labeling in the AMY, revealing the importance of an intact olfactory pathway for amygdaloid neurogenesis. VNOX increased anxiety-like behavior and altered social preference, but it did not affect social recognition memory in female voles. VNOX also reduced the percentage of BrdU-labeled cells that co-expressed the neuronal marker TuJ1 in the AMY, but not the DG. Together, our data indicate the importance of the olfactory pathway in mediating brain plasticity in the limbic system as well as its role in behavior. “
“Controllable/escapable tailshocks (ESs) do not produce the behavioral and neurochemical outcomes produced by equal yoked uncontrollable/inescapable tailshocks (ISs). The prelimbic cortex

is known to play a key role in mediating the protective effects of control. The concepts of act/outcome learning and control seem similar, and act/outcome learning is mediated by a circuit involving the prelimbic cortex and posterior dorsomedial striatum (DMS). Thus, we tested the involvement of the DMS in the protective effect of ES, in rats. First, we examined Fos immunoreactivity in both the DMS and dorsolateral striatum (DLS) after ES and yoked IS. We then investigated the effect of blocking DMS or DLS N-methyl-d-aspartate receptors with the specific antagonist D-(-)-2-amino-5-phosphopentanoic acid (D-AP5) on the release of dorsal raphe nucleus serotonin (5-HT) during ES, as well as on the level of anxiety produced by the ES experience 24 h later.

According to the sequencing result of the PCR products amplified by the primers S5un30 and S3un30, four specific primers

were designed: SP1: 5′-TTACTATCAATGTCTATAGGAGTAC-3′; SP2: 5′-AGCTGATCCTGGACCAGGCATAGC-3′; SP3: 5′-CATCTATGAATGGTCCACAAAATG-3′; and SP4: 5′-CGCTCGATCTGGCGGAGTGTATG-3′ were nested, respectively. The Son-PCR reactions (50 μL) were performed with 0.25 mmol L−1 of dNTP, 10 pmol of primer, and 2 U of Taq DNA polymerase. The DNA template of the primary reaction consisted of 20 ng of genomic DNA. The secondary reaction consisted of 2 μL of a 1 : 50 dilution of the first reaction. Following one denaturation step (3 min at 94 °C), the Selleck Everolimus reactions consisted of five cycles of amplification [30 s at 94 °C, 1 min at 62 or 66 °C (depending on the Tm of the

primers), 2.5 min at 72 °C], followed by one ramping step (30 s at 94 °C, 3 min at 29 °C, 3-min ramp to 72 °C, 2.5 min at 72 °C) and 60 (primary reaction) and 30 (secondary) new amplification cycles (10 s at 94 °C, 1 min at 62 or 66 °C, 2.5 min at 72 °C). The reaction ended with a final elongation step of 7 min at 72 °C. The final amplification products were ligated into the cloning vector: pMD18-T. The ligation reaction was carried out overnight at 4 °C in a 0.5-mL tube containing 1 μL pMD 18-T vector, 1 μL T4 DNA ligase, 3 μL PCR products, and 5 μL ligation buffer. Using the EZNA™ Gel Extraction Kit, an approximate 2.0-kb DNA product was purified from the plasmid containing the full-length sequence of the cry30Fa1 gene, digested with NcoI/XhoI, and inserted into multiple cloning sites of the expression vector Omipalisib pET-22b to generate the recombinant expression construct pET22-cry30Fa. The insert sequence and its reading frame were confirmed by the NcoI/XhoI digestion and DNA sequence analysis. The pET22-cry30Fa was transformed into E. coli BL21. Transformants were cultured overnight in 100 mL of LB medium with 100 μg ampicillin mL−1 at 37 °C, subcultured into a fresh medium (the volume ratio of 1 : 100)

for 6 h, and then induced with 1 mM isopropyl-β-d-thiogalactopyranoside P450 inhibitor (IPTG) for 4–6 h. Cells were harvested and resuspended in lysis buffer, sonicated, and centrifuged. The pellets were washed in order with 10 mL of 0.5 M NaCl and 2% Triton three times, 10 mL of 0.5 M NaCl five times, and 10 mL of double-distilled water two times. After centrifugation, at 9600 g for 10 min, the pellets were diluted and used for SDS-PAGE, which was performed using the procedure described by Sambrook et al. (2002). The resulting supernatant was loaded, at a flow rate of 100 μL min−1, onto a Sepharose CL-4B column precharged with Ni2+-chelated His-Bind resin (Qiagen). The column was washed with about 20 mL of wash buffer (50 mM Na2HPO4/NaH2PO4, 300 mM NaCl, 8 M urea, 20 mM imidazole). Proteins were then eluted with about 5 mL of elution buffer (50 mM Na2HPO4/NaH2PO4, 300 mM NaCl, 8 M urea, 500 mM imidazole).

To analyze the activity and specificity of the different OM cytochromes, we compared electron transfer to metals

and an anode surface. The reduction of an anode is as surface limited as the Bleomycin purchase reduction of an insoluble metal. However, anode reduction experiments can provide an additional set of information due to the possibility to change the rate of electron abstraction from the anode surface and thus the potential. The reduction experiments conducted showed that MtrCstrep and MtrFstrep could partly rescue the ΔOMC phenotype, while the production of other OM cytochromes resulted only in minor effects, if at all. A central role of MtrC in metal reduction is in agreement with earlier results (Beliaev et al., 2001; Myers & Myers, 2001) and might reflect the recently discovered capability of a complex of MtrC, with the β-barrel protein MtrB and the decaheme cytochrome MtrA, to

transport electrons over a liposome membrane and hence most probably also over the OM of S. oneidensis cells (Hartshorne et al., 2009). mtrF is part of a gene cluster that includes with mtrD and mtrE genes that are highly selleck screening library similar to mtrA and mtrB (McLean et al., 2008). We could show that MtrFstrep is a functional reductase that has, under several conditions, an even accelerated activity compared with MtrCstrep. McLean et al. (2008) speculate that the mtrDEF gene cluster could encode a reductase that is active under oxic or suboxic conditions and might have a function in Cobimetinib clinical trial reduction-based detoxification of radionuclides. The experiments presented here underline at least that MtrF is a reductase that could have this hypothetical function. The relative reduction activities of MtrFstrep compared with MtrCstrep follow the same pattern for all electron acceptors, except for an electrode in an MFC. Here, the LCD of MtrFstrep-producing cells is only 46% compared with the LCD achieved with MtrCstrep-producing cells. Therefore, we hypothesize that MtrFstrep might be not as well connected to the periplasmic electron pool, which could be due to

a reduced capability of forming a complex with MtrA and MtrB. This interprotein electron transfer might not be rate limiting under mineral-reducing conditions, but could become important when a certain current is applied to the MFC. OmcA production did not lead to accelerated reduction rates compared with the ΔOMC mutant in ferric iron reduction assays. This effect does not seem to be due to the reported partial mislocalization of OmcA in a ΔmtrC mutant (Myers & Myers, 2001) since proteinase K assays clearly demonstrated the surface exposure of OmcA in the ΔOMC mutant. OmcA is part of the core proteins that can be found in ferric iron-reducing S. oneidensis cells (Shi et al., 2007). We hypothesize that OmcA is an in vivo ferric iron reductase that is dependent on electron transport by another OM cytochrome. This cytochrome would most probably be MtrC.

BMC endocrine disorders 2013; 13: 1–12. Nicola Gray1, Janet McDonagh2, Kevin Harvey3, Julie Prescott4, Karen Shaw5, Felicity Smith6, David Terry2, Kate Fleck7, Rachel Roberts8 1Green Line Consulting SAHA HDAC in vivo Limited, Manchester, UK, 2Birmingham Children’s Hospital NHS Foundation Trust, Birmingham, UK, 3University of Nottingham, Nottingham, UK, 4University of Central Lancashire, Preston, UK, 5University of Birmingham, Birmingham, UK, 6UCL School of Pharmacy,

London, UK, 7Arthritis Care, Belfast, UK, 8Pharmacy Research UK, London, UK The aim of this abstract is to describe the psychosocial context of medicine-taking for young people living with arthritis Family partnerships, relationships Cobimetinib with peers, and the demands of school life all impact significantly on the medicine-taking practices of young people To be able to provide meaningful advice and services for young people, pharmacists must consider the psychosocial contextual factors in young people’s lives that influence medicine-taking Better communication with the local rheumatology team, and an understanding of their

prescribing practice, would equip pharmacists to support young people with JIA Young people with juvenile idiopathic arthritis (JIA) may have complex medicine routines – including injections – that don’t fit with their ideas of ‘normal’1. Their medicines have to be taken even when they feel well – to keep them that way. Pharmacists may be able to support medicines optimisation for this population as part of the arthritis provider team. Consultation opportunities are afforded by Medicines Use Review (MUR) and the New Medicines Service in England, the Chronic

Medication Service in Scotland, and MUR in Wales. The aim of this abstract is to describe the psychosocial context of medicine-taking for young people living with arthritis. Young people (aged 11–15) with arthritis – recruited from adolescent rheumatology clinics at Birmingham Children’s Hospital NHS Foundation Trust, England – wrote blogs on a bespoke ‘Arthriting’ website. These anonymised personal blogs included thoughts about identity, their arthritis condition, medication, and the use of health Amisulpride services. Young people could contribute blogs over a 2-month period from their registration with the site. Qualitative data were subjected to directed content analysis2, which allowed us to pursue pre-existing themes of interest, but also allowed new themes to emerge. Ethical approval was obtained from Coventry & Warwickshire NRES REC. Twenty-one young people took part, collectively contributing 161 blog entries. Contextual factors described in the blogs included the family, relationships with peers, and school life. Young people had help with many aspects of medication use; mothers often helped with getting supplies, setting routines, and giving medication.

The authors would like to thank Ms Maiko Uezaki for her assistance with MEG measurement. This work was supported by a Japan Society for the Promotion of Science (JSPS) KAKENHI Grant-in-Aid for Young Scientists (B) (24730618) to K.O. and by the Special Coordination Fund for Promoting Science and Technology to C.F.A. from the Ministry of Education, Culture, Sports, Science and

Technology (MEXT) of Japan. The authors have no conflict of interest to declare. Abbreviations IPL inferior parietal lobe ISI inter-stimulus interval L loud MEG magnetoencephalography MNI Montreal Neurological R428 in vivo Institute ROI region of interest S soft STG superior temporal gyrus “
“Word recognition research with alphabetical scripts has revealed a facilitatory neighborhood size effect, whereby naming of words find more with more orthographic neighbors is faster than that of words with fewer neighbors. Preliminary behavioral evidence in Chinese revealed both facilitatory and inhibitory neighborhood size effects,

depending on whether there are higher-frequency neighbors (HFNs) than the target. This functional magnetic resonance imaging study examined the neural substrates of the neighborhood size effect with silent naming. Neighborhood size and the HFN factor were factorially manipulated. Behavioral results replicated previous findings showing that larger neighborhood size facilitated naming in the absence of HFNs, but inhibited naming in their presence. Imaging results identified greater activation in the left middle frontal gyrus for small than larger neighborhood size, and bilateral inferior frontal activations for the with-HFN condition as compared with the without-HFN condition. Critically, there was an interaction in the right middle occipital gyrus showing greater activation for large than for small neighborhood size in the absence of HFNs but no neighborhood

Montelukast Sodium size effect in their presence. The results support a proposal that, in addition to a facilitatory contribution from orthographic activation of neighborhoods, naming is also affected by whether there are higher-frequency neighbors, particularly in scripts with deep orthography, where orthographically similar words can be pronounced very differently. “
“Most default mode network (DMN) studies in patients with amnestic mild cognitive impairment (aMCI) and Alzheimer’s disease (AD) are based on the comparison of only two groups, namely patients and controls. Information derived from comparing three groups, normal, aMCI and AD, simultaneously may lead us to better understand the progression of dementia. The purpose of this study was to evaluate functional connectivity of DMN in the continuum from normal through aMCI to AD. Differences in functional connectivity were compared between the three groups using independent component analysis.

A functional general stress response is probably needed in the manure-amended soil environment and nutrient availability is not the most limiting factor. This has also been shown for the plant pathogenic bacterium Pseudomonas putida (Ramos-González & Molin, 1998). However, deletion and complementation studies should provide more evidence for the role of RpoS in the survival of E. coli O157 in manure-amended soil. In addition, the conditions for survival in non-amended soil might be completely different and the role of RpoS should be considered accordingly. Variation in

rpoS alleles has been observed among E. coli O157 isolates and it remains unclear which environment gives Olaparib manufacturer rise to and selects for rpoS mutants (Waterman & Small, 1996; Parker et al., 2012). None of the E. coli O157 strains isolated from the environment (from feral pig, river water, cow and pasture soil) and linked to the 2006 spinach-associated outbreak in the United

States showed mutations in the rpoS gene (Parker et al., 2012). In contrast, 3/3 clinical and 2/5 spinach isolates possessed mutations in the rpoS gene, produced TSA HDAC lower levels of RpoS, showed decreased levels of rpoS-regulated genes, and showed impaired phenotypic resistance to low pH, osmotic stress and oxidative stress. Parker et al. (2012) suggested that bagged spinach could provide a niche for the rise and selection of rpoS mutants and that these mutants are merely maintained during passage through the human host. However, this suggestion is challenged by gene expression studies showing clear up-regulation of rpoS when associated with (injured) lettuce tissue, implying the need for a functional general stress response (Carey et al., 2009; Kyle et al., 2010; Fink et al., 2012). As the bovine intestine forms the principal reservoir of E. coli O157 and humans can be considered

a transient host with distinct gastrointestinal conditions, it Bumetanide was hypothesized that the human gut could provide a niche for the selection of rpoS mutants. Sequencing the structural part of the rpoS gene of 73 bovine, 29 food (23 meat, one lettuce and five endive isolates) revealed a skewed distribution of mutants among the different isolation sources. Bovine and food isolates had low numbers of mutants (0/73 and 2/29, respectively), while a relatively high number of mutants was observed among human isolates (28/85) (Table 2). A strong LSPA-6-specific distribution of rpoS(A543C) among the isolates was observed, with 100% of lineage I possessing rpoS(543A) whereas 100% of the lineage II strains had rpoS(543C). Lineage I/II were either rpoS(543A) or rpoS(543C): bovine strains, 44.8% rpoS(543A) and 55.2% rpoS(543C); food strains, 26.7% rpoS(543A) and 73.3% rpoS(543C); human clinical strains, 49.2% rpoS(543A) and 50.8% rpoS(543C). This is in agreement with earlier findings that lineage I/II is genetically more diverse than lineage I and II (Bono et al., 2012; Eppinger et al., 2012).

2). FK77 (ΔbceRS), MM02 (ΔbceAB) and MM03 (ΔvraDE) strains reduced their resistance level, revealing hetero-resistance to bacitracin. MM03

in particular showed a significant reduction compared with two other mutants, FK77 and MM02. In wild-type MW2 strain, the expression of bceA and vraD was rapidly induced by the addition of bacitracin into the medium (Fig. 3). This induction was found to occur after 5 min of bacitracin exposure. The level of vraD transcript was more than 100-fold that of the wild type. Interestingly, a low concentration of bacitracin (0.5, 1 μg mL−1) significantly induced the expression of two genes (bceA and vraD) until 15 min, after which the expression of both decreased. In contrast, bacitracin (above 8 μg mL−1) continued Sotrastaurin clinical trial to induce these two genes even after 15 min. The level of the TCS (bceR) transcript itself did not increase by addition of bacitracin after 30 min, but slightly increased after 60 min (data not shown). The level of vraF transcript was not increased

by bacitracin (data not shown). The expressions of bceA and vraD in MM08 (ΔbceS) were not induced by addition of bacitracin, while the induction of their expressions by bacitracin were observed in the strain (MM09) which complemented with the gene for bceS in MM08 (Fig. 3). Also, in FK77 (ΔbceRS) strain, induction of the expression of bceA and vraD upon the addition of bacitracin was completely inhibited. XL184 order In this study, we identified one uncharacterized TCS (MW2545-44) that has the ability to sense bacitracin and positively regulates two transporters responsible for bacitracin resistance. Interestingly, this TCS regulates not only a downstream gene coding for ABC transporter (MW2543-42), but also another transporter (MW2620-2621) known as vraDE, which is located separately

from the genes coding for the TCS. These two ABC transporters, especially VraDE, showed a high similarity with B. subtilis ABC transporter Etomidate BceAB responsible for bacitracin efflux (Ohki et al., 2003). In addition, similar transporters showing homology with BceAB have been reported in several Gram-positive bacteria, such as BcrAB of B. licheniformis (Podlesek et al., 1995), BcrAB of Enterococcus faecalis (Manson et al., 2004) and MbrAB of Streptococcus mutans (Tsuda et al., 2002). This type of ABC transporter is considered to pump out bacitracin to the exterior of the cell directly. Upstream of bceAB in B. subtilis, the gene coding for TCS, known as BceRS, is located (Ohki et al., 2003; Rietkötter et al., 2008). BceRS has been demonstrated to sense bacitracin and induce the expression of BceAB, leading to resistance to bacitracin. Like bceRSAB in B. subtilis, the results in this study showed that S. aureus has the same system, so we designated this TCS as BceRS (MW2545-2544) and the downstream transporter as BceAB (MW2543-2542). In S. aureus, another gene, MW2546, was closely located upstream of bceRS, speculating three genes were in a same operon (Fig. 1).

Long-term exposure to DHT or E2 did not result in a shortened free-running circadian period when administered at 2–7 or 14–19 months of age. However, E2 treatment from 7 to 12 months of age decreased the free-running circadian period in castrated males. This result was replicated in a subsequent experiment in which E2 treatment was limited to 8–12 months of age. E2 treatment at 7–12 months of age had no effect on the free-running circadian period in ovariectomized females. Thus, there appears to be

a post-pubertal sensitive period for sexual differentiation of the circadian system of selleckchem degus, during which E2 exposure decreases the free-running circadian period in males. These data demonstrate that gonadal hormones can act during adolescent development to permanently alter the circadian system. “
“The frontal eye field (FEF), in the prefrontal cortex, participates in the transformation of visual signals into saccade motor commands and in eye–head gaze control. The FEF is thought to show eye-fixed visual codes in head-restrained monkeys, but it is not known how it transforms these inputs into spatial codes for head-unrestrained gaze commands. Here, we tested if the FEF influences desired gaze Cisplatin purchase commands within a simple eye-fixed frame, like the superior colliculus (SC), or in more complex egocentric frames like the supplementary eye fields (SEFs).

We electrically stimulated 95 FEF sites in two head-unrestrained monkeys to Megestrol Acetate evoke 3D eye–head

gaze shifts and then mathematically rotated these trajectories into various reference frames. In theory, each stimulation site should specify a specific spatial goal when the evoked gaze shifts are plotted in the appropriate frame. We found that these motor output frames varied site by site, mainly within the eye-to-head frame continuum. Thus, consistent with the intermediate placement of the FEF within the high-level circuits for gaze control, its stimulation-evoked output showed an intermediate trend between the multiple reference frame codes observed in SEF-evoked gaze shifts and the simpler eye-fixed reference frame observed in SC-evoked movements. These results suggest that, although the SC, FEF and SEF carry eye-fixed information at the level of their unit response fields, this information is transformed differently in their output projections to the eye and head controllers. “
“Polysialic acids are widely distributed in neuronal tissue. Due to their position on glycoproteins and gangliosides on the outer cell membranes and anionic nature, polysialic acids are involved in multiple cell signaling events. The level of sialylation of the cellular surface is regulated by endogenous neuraminidase (NEU), which catalyses the hydrolysis of terminal sialic acid residues.

No bands were observed when the blots were hybridized with a probe corresponding to hynSL, confirming that the genes encoding the hydrogenase were deleted (Fig. 3c). When the

blots were rehybridized with a probe to the kanamycin resistance gene, bands of the expected size were identified, confirming that double recombination had occurred and that the KmR cassette had replaced the hydrogenase genes (Fig. 3d). Mutants confirmed by Southern blot analysis were also found to lack hydrogenase activity in an in vitro hydrogen evolution assay (Fig. 3e). To complement the PW440 hydrogenase mutant, a plasmid, pPW438, containing the wild-type AltDE hydrogenase gene cluster was conjugated into the mutant and a single recombination event in the mutant was selected by resistance to the antibiotics chloramphenicol and

kanamycin. PCR amplification Proteasome inhibitor of hynSL confirmed that the complemented strain contained a copy of the wild-type hydrogenase genes (Fig. 3b). Hydrogenase activity measured by in vitro hydrogen evolution assay with cell extracts indicated that the complemented strain, PW438/PW440, regained hydrogenase activity to almost wild-type levels (Fig. 3e). To learn more about the physiology of A. macleodii, we investigated the ability of AltDE to grow under various CDK and cancer conditions. While AltDE grew well in the complete medium (marine broth) under aerobic conditions, growth under anaerobic conditions was inhibited unless nitrate was added to the medium as an electron acceptor (Fig. 4a). No growth was observed when sulfate was provided as an electron acceptor (data not shown). When grown in a complete medium with nitrate under anaerobic conditions containing 3% H2, no differences in the growth rate were observed between the wild type and the hydrogenase mutant strain, PW440 (Fig.

4a). Strains U7, U8, U10, and U12 were isolated by Sass et al. (2001) in the Urania Basin at a depth of 3500 m, where the chloride concentration was measured to be between 2.5 and 3.0 M. Thus, we tested the ability of A. macleodii to grow in the presence of additional salt. When the complete marine medium was supplemented with an additional 2 M NaCl, growth was slowed, but still detectable (Fig. 4b and c). This slower growth in the high-salt medium was detected when grown either aerobically or anaerobically (nitrate was supplied as the electron acceptor) Glycogen branching enzyme (Fig. 4b and c). As expected, no growth occurred in minimal seawater media (Fig. 4b and c). No significant differences in the growth rates could be observed between the wild type and the hydrogenase mutant strains of AltDE under all growth conditions tested (Fig. 4b and c). Thus, the presence of the hydrogenase does not appear to affect growth rate in the presence of 3% H2 in a complete medium. The fact that no growth was detected in minimal seawater in the presence of 3% H2 is consistent with the designation of A. macleodii as a chemoheterotroph that requires fixed carbon sources.

Mass spectra were acquired by a Finnigan™ LCQ™ DECA ion trap instrument. An ionization device was used for sample analyses (sheath gas: 80 mL min−1, auxiliary gas: 20 mL min−1, spray voltage: 5 kV, capillary temperature: 300 °C, capillary voltage: 46 kV, and tube lens: −60 kV). The Xcalibur 2.0

SR2 software (copyright Thermo Electron Corporation 1998–2006) was used. Morphological and cultural studies of the most productive isolate containing the ts gene, SBU-16, including conidial morphology, the mechanism this website of conidia production, and growth characteristics on PDA, potato-carrot agar (PCA), and on the firm base of an alfalfa stem were carried out according to Simmons (2001). The isolate of SBU-16 was grown on the media in a culture chamber under selleckchem a 10-h photoperiod provided by 56 W cool-white fluorescent lamps (Philips Master, Holand) at 22 °C. Anamorph and telomorph populations were examined at 4–5 days and 2–6 weeks, respectively. The size and morphology of 100 mature conidia and 50 conidiophores

in lactic acid were recorded by light microscopy at 100× magnification and photographed. A total of 25 isolates separated from the inner bark of T. baccata were screened for the presence of the ts gene. Based on the conserved region of the ts gene, the specific primers were designed and synthesised for the amplification of the core DNA fragment of ts from 25 isolated endophytic fungi. Following PCR amplification, a 334-bp product was obtained. Of 25 isolates, 4 (SBU-16, SBU-17, SBU-69 and SBU-71) showed PCR positive for the conserved sequence of the ts gene (Fig. 1). Taxol and 10-DAB III were extracted from culture filtrates and mycelia of the four ts PCR positive fungi and then analyzed Non-specific serine/threonine protein kinase by HPLC-DAD. Under the same analysis conditions, the samples containing chemical reference substances of 10-DAB III and taxol were also compared with fungal extracts (Fig. 2). Further convincing evidence for the identity of 10-DAB III and taxol was obtained by high-performance

liquid chromatography-mass spectrometry (LC-MS). Characteristically, standard 10-DAB III and taxol yielded both an [M + H]+ peak at a molecular weight of 854 and an [M + Na]+ peak at a molecular weight of 876, respectively (see Fig. 3a and b). By comparison, fungal taxol also produced peaks, [M + H]+ at m/z 854 and [M + Na]+ at m/z 876. The peaks corresponding to taxol exhibited mass-to-charge (m/z) ratios corresponding to the molecular ions (M + H)+ of standard taxol (at 854), confirming the presence of taxol in the fungal extracts. It was evident that taxol was much more complex because its molecular weight (from high-resolution mass spectrometry) was 854, which corresponds to a molecular formula of C47H51NO14 as reported earlier (McClure & Schram, 1992). The results of the quantification analysis among the four ts PCR positive isolates showed that SBU-16, which was isolated for the first time in our laboratory, produces taxol (6.9 ± 0.2 μg L−1) and its intermediate compound, 10-DAB III (2.