Seventeen studies were analysed using activation likelihood estimation. Trait stuttering was characterised by the well-known rightward shift in lateralization for language and speech areas. State stuttering

revealed a more diverse pattern. Abnormal activation of larynx and lip motor cortex was common to the two analyses. State stuttering was associated with overactivation in the right hemisphere larynx and lip motor cortex. Trait stuttering was associated with overactivation of lip motor cortex in the right hemisphere but underactivation of larynx motor cortex in the left hemisphere. These results support a large literature highlighting laryngeal and lip involvement in the symptomatology of stuttering, and disambiguate two possible sources of activation in neuroimaging studies of persistent developmental stuttering. “
“The volatile anesthetic ABT-199 ic50 GSI-IX cell line sevoflurane, which is widely used in pediatric

surgery, has proposed effects on GABAA receptor-mediated extrasynaptic tonic inhibition. In the developing striatum, medium-sized spiny projection neurons have tonic GABA currents, which function in the excitatory/inhibitory balance and maturation of striatal neural circuits. In this study, we examined the effects of sevoflurane on the tonic GABA currents of medium spiny neurons in developing striatal slices. Sevoflurane strongly increased GABAA receptor-mediated tonic conductance at postnatal days 3–35. The antagonist oxyclozanide of the GABA transporter-1, 1-[2-[[(diphenylmethylene)imino]oxy]ethyl]-1,2,5,6-tetrahydro-3-pyridinecarboxylic acid hydrochloride further increased tonic GABA conductance during the application of sevoflurane, thereby increasing the total magnitude of tonic currents. Both GABA (5 μm) and 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridine-3-ol hydrochloride, the δ-subunit-containing GABAA receptor agonist, induced tonic GABA currents in medium spiny neurons but not in cholinergic neurons. However, sevoflurane additively potentiated the tonic GABA currents in both cells. Interestingly, 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridine-3-ol hydrochloride-sensitive neurons made a large current response to sevoflurane, indicating

the contribution of the δ-subunit on sevoflurane-enhanced tonic GABA currents. Our findings suggest that sevoflurane can affect the tone of tonic GABA inhibition in a developing striatal neural network. “
“Here we report early cross-sensory activations and audiovisual interactions at the visual and auditory cortices using magnetoencephalography (MEG) to obtain accurate timing information. Data from an identical fMRI experiment were employed to support MEG source localization results. Simple auditory and visual stimuli (300-ms noise bursts and checkerboards) were presented to seven healthy humans. MEG source analysis suggested generators in the auditory and visual sensory cortices for both within-modality and cross-sensory activations.

Expert blood film microscopy remains the mainstay in the diagnosis of malaria but molecular tools may provide important additional information. Importantly, this case emphasizes the necessity of routine checkups of parasitemia following

treatment and whenever indicated by the clinical course. We thank S. Zander for excellent technical assistance. The authors state DNA Synthesis inhibitor that they have no conflicts of interest to declare. “
“Objective Noninvasive tests that can be used in place of liver biopsy to diagnose fibrosis have major limitations. They either leave a significant proportion of patients without a definitive diagnosis or produce inaccurate results. Moreover, the performance of these tests is lower in HIV/hepatitis C virus (HCV) coinfection. Against this background, PF-562271 we examined the utility of serum matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 1 (TIMP-1) measurements in combination with routine clinical data to predict fibrosis in HIV/HCV-coinfected

patients. Methods Patients with a liver biopsy who had not received anti-HCV therapy were included in the study. A model including variables independently associated with fibrosis was constructed. Diagnostic accuracy was determined by measuring the area under the receiver operating characteristic curve (AUROC). Positive (PPV) and negative (NPV) predictive values were calculated. Results Ninety patients were included in the study. Aspartate aminotransferase (AST), platelet count and MMP-2 were predictors of significant MTMR9 fibrosis (F≥2) and cirrhosis (F4). A score constructed using these variables yielded an AUROC of 0.76 for F≥2 and 0.88 for F4. Score cut-offs detected (value ≥3.5) and excluded (value ≤1.5) F≥2 with a PPV of 87% and an NPV of 88%. Thirty-one patients

(34%) were correctly diagnosed using these cut-offs, with four (13%) incorrect classifications. Cirrhosis was excluded with a certainty of 98% and diagnosed with a probability of 83%. Two (17%) of 12 patients were misclassified as having cirrhosis. The AST to platelet count index and MMP-2 levels were sequentially applied to detect F≥2. Forty-one patients (46%) were identified with this approach, with six (15%) misclassifications. Conclusion MMP-2 levels can be used in combination with AST and platelet count to aid the diagnosis of liver fibrosis in HIV/HCV-coinfected patients. The extent of liver fibrosis has prognostic and management implications in chronic hepatitis C. The diagnosis of fibrosis has traditionally relied on liver biopsy. However, this procedure is invasive, limited because of variability issues [1,2] and difficult to apply sequentially. Because of these issues, noninvasive tests that can be used in place of liver biopsy are needed.

e. PBAD promoter repressed (data not shown). Since R.PamI and the commercial endonuclease NcoI show

some amino acid sequence similarity, we tested whether or not these enzymes exhibit the same sequence specificity. For this purpose a His-tagged version, R.PamI(His)6, was expressed and purified, and the optimal conditions for DNA cleavage by the recombinant protein were determined (different temperatures and buffers were tested). As shown in Fig. 3, the restriction profiles of λ DNA cleaved by the two REases were identical, although the optimal temperature for R.PamI activity was lower (30 °C; the same as for growth of the host strain JCM 7686) than that for NcoI (37 °C). To determine the cleavage sequence of R.PamI, plasmid pET28b, containing a single NcoI site (5′-C▼CATG▲G-3′), NVP-BGJ398 cost was used. This plasmid was cleaved with R.PamI(His)6, the putative overhangs of the linearized DNA molecule were filled with Klenow DNA polymerase and it was Nutlin-3a cost recircularized by blunt-end ligation. DNA sequencing revealed modification of the NcoI site (5′-CCATGCATGG-3′; duplicated CATG sequence is underlined), which unequivocally proved that R.PamI(His)6 and NcoI exhibit the same specificity and both cleave their target sequence after the first cytosine. The plasmid pAMI7 contains six NcoI sites, but when isolated from P. aminophilus JCM 7686, this DNA was completely resistant

to cleavage by NcoI (data not shown). According to REBASE (Roberts et al., 2010), NcoI is sensitive to m5C methylation of the first cytosine in its recognition sequence (CCATGG), but there are no data concerning the sensitivity of this enzyme to methylation

of the second cytosine. To determine which C within the PamI recognition sequence is the triclocarban target for MTase M.PamI, we used the construct pACYC184/MRW as a substrate DNA. The oligonucleotide duplex shown in Fig. 4 was inserted into the plasmid pACYC184. In this 12-bp sequence, two PamI recognition sequences overlap one BsuRI site (ccatGGCCatgg), and NlaIII sites (CATG) are contained within each PamI recognition sequence (cCATGgcCATGg) (Fig. 4b). If M.PamI methylates the first cytosine in its recognition sequence (CCATGG), the BsuRI endonuclease cannot cut at the site between the two PamI sites (BsuRI is not sensitive to methylation of the second cystosine). On the other hand, if the second cytosine is methylated to m5C, the modification will affect NlaIII digestion. pACYC184/MRW DNA was isolated from an E. coli TOP10 strain that also carried plasmid pAMI702. Figure 4a shows that this preparation of pACYC184/MRW was completely digested with NlaIII (absence of uncleaved 2107-bp band comprised of 1718- and 278-bp fragments), but cleavage with BsuRI was partially inhibited at the position 5061 resulting in the presence of an uncleaved 1900-bp band (777- plus 1123-bp fragments). This experiment revealed that M.

e. PBAD promoter repressed (data not shown). Since R.PamI and the commercial endonuclease NcoI show

some amino acid sequence similarity, we tested whether or not these enzymes exhibit the same sequence specificity. For this purpose a His-tagged version, R.PamI(His)6, was expressed and purified, and the optimal conditions for DNA cleavage by the recombinant protein were determined (different temperatures and buffers were tested). As shown in Fig. 3, the restriction profiles of λ DNA cleaved by the two REases were identical, although the optimal temperature for R.PamI activity was lower (30 °C; the same as for growth of the host strain JCM 7686) than that for NcoI (37 °C). To determine the cleavage sequence of R.PamI, plasmid pET28b, containing a single NcoI site (5′-C▼CATG▲G-3′), selleckchem was used. This plasmid was cleaved with R.PamI(His)6, the putative overhangs of the linearized DNA molecule were filled with Klenow DNA polymerase and it was Avasimibe clinical trial recircularized by blunt-end ligation. DNA sequencing revealed modification of the NcoI site (5′-CCATGCATGG-3′; duplicated CATG sequence is underlined), which unequivocally proved that R.PamI(His)6 and NcoI exhibit the same specificity and both cleave their target sequence after the first cytosine. The plasmid pAMI7 contains six NcoI sites, but when isolated from P. aminophilus JCM 7686, this DNA was completely resistant

to cleavage by NcoI (data not shown). According to REBASE (Roberts et al., 2010), NcoI is sensitive to m5C methylation of the first cytosine in its recognition sequence (CCATGG), but there are no data concerning the sensitivity of this enzyme to methylation

of the second cytosine. To determine which C within the PamI recognition sequence is the Amylase target for MTase M.PamI, we used the construct pACYC184/MRW as a substrate DNA. The oligonucleotide duplex shown in Fig. 4 was inserted into the plasmid pACYC184. In this 12-bp sequence, two PamI recognition sequences overlap one BsuRI site (ccatGGCCatgg), and NlaIII sites (CATG) are contained within each PamI recognition sequence (cCATGgcCATGg) (Fig. 4b). If M.PamI methylates the first cytosine in its recognition sequence (CCATGG), the BsuRI endonuclease cannot cut at the site between the two PamI sites (BsuRI is not sensitive to methylation of the second cystosine). On the other hand, if the second cytosine is methylated to m5C, the modification will affect NlaIII digestion. pACYC184/MRW DNA was isolated from an E. coli TOP10 strain that also carried plasmid pAMI702. Figure 4a shows that this preparation of pACYC184/MRW was completely digested with NlaIII (absence of uncleaved 2107-bp band comprised of 1718- and 278-bp fragments), but cleavage with BsuRI was partially inhibited at the position 5061 resulting in the presence of an uncleaved 1900-bp band (777- plus 1123-bp fragments). This experiment revealed that M.

“
“Immunocytochemistry

shows that purinergic receptors (P1Rs) type A1 and A2A (A1R and A2AR, respectively) are present in the nerve endings at the P6 and P30 Levator auris longus (LAL) mouse neuromuscular junctions (NMJs). As described elsewhere, 25 μm adenosine reduces (50%) acetylcholine release in high Mg2+ or d-tubocurarine paralysed muscle. We hypothesize that in more preserved neurotransmission machinery conditions (blocking the voltage-dependent sodium channel of the muscle www.selleckchem.com/products/gsk1120212-jtp-74057.html cells with μ-conotoxin GIIIB) the physiological role of the P1Rs in the NMJ must be better observed. We found that the presence of a non-selective P1R agonist (adenosine) or antagonist (8-SPT) or selective modulators of A1R or A2AR subtypes (CCPA and DPCPX, or CGS-21680 and SCH-58261, respectively) does not result in any changes in the evoked release. However, P1Rs seem to be involved in spontaneous release (miniature endplate potentials MEPPs) because MEPP frequency is increased by non-selective block but decreased by non-selective stimulation, with A1Rs playing the main role. We assayed the role of P1Rs in presynaptic short-term plasticity during imposed synaptic activity (40 Hz for 2 min of supramaximal stimuli). Depression is reduced by micromolar adenosine but increased by blocking P1Rs with 8-SPT. Synaptic depression is not affected by the presence of selective A1R and A2AR modulators, which suggests that both receptors

need to collaborate. Thus, A1R and A2AR might have no real effect on neuromuscular transmission in resting conditions. However, these receptors can conserve resources by limiting spontaneous quantal leak of Gefitinib cost acetylcholine and may protect synaptic function by reducing the magnitude of depression during repetitive activity. “
“Morphine remains one of the most potent analgesic compounds used to control chronic pain despite its known adverse effects. It binds to the opioid receptors mu, delta and kappa, which are involved in aspects of neuronal fate such as cell proliferation, neuroprotection and neuronal differentiation.

However, the effect of morphine on these processes is controversial and in vitro studies, as well as in vivo Fenbendazole studies on adults and neonates in mammalian models, have not been able to clarify the diverse roles of morphine in the central nervous system. We have used zebrafish embryos to determine in vivo how morphine affects neuronal fate and opioid receptor gene expression and to elucidate if there is a link between these processes. Our results show that at 24 and 48 h post fertilization (hpf) morphine enhances cell proliferation, although it has opposing effects as an inducer of neuronal differentiation at these two stages, increasing the number of certain neuronal populations at 24 hpf and decreasing it at 48 hpf. The present study also demonstrates that in 24-hpf embryos morphine acts as a neuroprotector against glutamate damage in motor neurons and Pax-6-positive neurons.

Heptachlor diol and 1-hydroxy-2,3-epoxychlordene were produced in these fungal cultures as metabolites, suggesting that the hydrolysis and hydroxylation reaction occur in the epoxide ring and in position 1 of heptachlor epoxide, respectively. Over the past few decades, the presence of organochlorine pesticides (OCPs) in the environment has been of great concern due to their persistent, long-range transportable nature and toxic biological effects. Heptachlor is an OCP that was used extensively in the developed world throughout the 1960s and 1970s, mainly against termites and soil insects.

Some developed countries banned or restricted Tofacitinib order the production and usage of heptachlor in the 1970s because animal data suggested that it is carcinogenic in humans (World Health Organization, 1984). Nevertheless, some developing countries continue to use this

pesticide in both agriculture and public health programs because of its low cost and versatility in controlling various pests. Heptachlor has not been produced in Japan, but 1500 tons were imported between 1958 and 1972 (Murano et al., 2009). The Japanese government banned the use of heptachlor in 1972. Heptachlor Selleckchem Palbociclib is likely to remain in the soil for long periods of time (Huber, 1993), albeit at relatively low concentrations (parts per billion). Its reported representative field half-life is 250 days (Augustijn-Beckers et al., 1994). However, traces of heptachlor have been detected in soil even 14 and 16 years after application. A widespread reaction in the environment is heptachlor Florfenicol epoxidation to the more persistent heptachlor epoxide. Heptachlor and heptachlor epoxide are relatively hydrophobic compounds and therefore extensively adsorb onto soil particles, giving these compounds low bioavailability and mobility in soil. Several studies have reported elevated concentrations of heptachlor and heptachlor epoxide in surface water, sediment and soil samples from Asian countries including China, Japan and Thailand (Kim et al., 2007; Gao et al.,

2008; Poolpak et al., 2008). The first evidence that heptachlor is degraded by soil microorganisms came from the experiments of Miles et al. (1969). In their studies, heptachlor is metabolized by soil bacteria and fungi into many different products by many independent metabolic pathways. Heptachlor epoxide, chlordene, chlordene epoxide, 1-hydroxychlordene and 1-hydroxy-2,3-epoxychlordene were the products of the microbial degradation of heptachlor (Fig. 1). Currently, bioremediation conducted on a commercial scale utilizes bacteria; there have been few attempts to use white rot fungi. However, white rot fungi offer advantages over bacteria in the diversity of compounds they can oxidize (Pointing, 2001). These organisms are generally more tolerant to high concentrations of polluting chemicals than bacteria.

Total RNA was isolated using the SV RNA isolation kit (Promega) according to the manufacturer’s instructions. RT-PCR was performed modeling a previously published report (Suzuki et al., 2004). The following modifications were used. The affinity script multiple temperature cDNA synthesis kit (Stratagene) was

utilized along with a gene-specific primer P2 in first-strand synthesis from total RNA. The PCRx enhancer system (Invitrogen Life Technology) was used according to the manufacturer’s recommendation in the amplification step. The primers used were as follows: P1, 5′-CATGGCTCGCCGCGCTGTCG-3′; P2, 5′-CGCTGGTCGGCATAGAACTC-3′; P3, 5′-CAGCGATCGTCGGCGCATCG-3′; and P4, 5′-GACCAGGCTGTAGTCGCCGA-3′. As an initial step toward identifying the molecular determinants of the oxalic acid biosynthetic pathway in B. glumae, a transposon-mutagenized library (Nakata, 2002) NVP-LDE225 was screened for mutants in oxalate production. Cultures of individual colonies from the mutagenized library were grown in liquid media and the cultures were assayed for the presence of oxalic

acid using a colorimetric detection system. After screening approximately 3000 colonies, four mutants were identified that failed to produce and secrete detectable levels of oxalic acid into the media (Fig. 1a and b). These mutants were named Bod1. To assess whether the Bod1 knockout phenotype was due to the lack of the biosynthetic step, oxalic acid biosynthetic enzyme assays Rucaparib datasheet were conducted. Protein extracts were prepared from control and mutant cells and dialyzed to remove endogenous oxalic acid and other metabolites. Control extracts showed the ability to produce oxalic acid from oxaloacetate and acetyl-CoA in vitro, while all four mutants lacked this biosynthetic activity (Fig. 1c). Southern blot analysis was performed to determine whether the Bod1 mutants resulted from a single or multiple insertion events. Southern blots using different restriction enzymes revealed that each Bod1 mutant contained a single CHIR-99021 mw insertion in a restricted fragment of similar

size (data not shown). The results indicated that a single essential gene may have been affected. To determine whether this was indeed the case, the area flanking the insertion site of each Bod1 mutant was sequenced. DNA sequence analysis revealed that each Bod1 mutant was a result of an independent insertion event into the same ORF (Fig. 2a). The identified ORF was named obcA. A blast search, using the obcA sequence, against the GenBank database revealed three matches with significant homology. An identity of approximately 67% was found between obcA and an ORF from Burkholderia ubonensis and about 53% identity was found between the obcA sequence and an ORF found in two related human pathogenic bacteria: Burkholderia pseudomallei and Burkholderia mallei.

Total RNA was isolated using the SV RNA isolation kit (Promega) according to the manufacturer’s instructions. RT-PCR was performed modeling a previously published report (Suzuki et al., 2004). The following modifications were used. The affinity script multiple temperature cDNA synthesis kit (Stratagene) was

utilized along with a gene-specific primer P2 in first-strand synthesis from total RNA. The PCRx enhancer system (Invitrogen Life Technology) was used according to the manufacturer’s recommendation in the amplification step. The primers used were as follows: P1, 5′-CATGGCTCGCCGCGCTGTCG-3′; P2, 5′-CGCTGGTCGGCATAGAACTC-3′; P3, 5′-CAGCGATCGTCGGCGCATCG-3′; and P4, 5′-GACCAGGCTGTAGTCGCCGA-3′. As an initial step toward identifying the molecular determinants of the oxalic acid biosynthetic pathway in B. glumae, a transposon-mutagenized library (Nakata, 2002) ABT-199 was screened for mutants in oxalate production. Cultures of individual colonies from the mutagenized library were grown in liquid media and the cultures were assayed for the presence of oxalic

acid using a colorimetric detection system. After screening approximately 3000 colonies, four mutants were identified that failed to produce and secrete detectable levels of oxalic acid into the media (Fig. 1a and b). These mutants were named Bod1. To assess whether the Bod1 knockout phenotype was due to the lack of the biosynthetic step, oxalic acid biosynthetic enzyme assays RG7422 order were conducted. Protein extracts were prepared from control and mutant cells and dialyzed to remove endogenous oxalic acid and other metabolites. Control extracts showed the ability to produce oxalic acid from oxaloacetate and acetyl-CoA in vitro, while all four mutants lacked this biosynthetic activity (Fig. 1c). Southern blot analysis was performed to determine whether the Bod1 mutants resulted from a single or multiple insertion events. Southern blots using different restriction enzymes revealed that each Bod1 mutant contained a single Oxymatrine insertion in a restricted fragment of similar

size (data not shown). The results indicated that a single essential gene may have been affected. To determine whether this was indeed the case, the area flanking the insertion site of each Bod1 mutant was sequenced. DNA sequence analysis revealed that each Bod1 mutant was a result of an independent insertion event into the same ORF (Fig. 2a). The identified ORF was named obcA. A blast search, using the obcA sequence, against the GenBank database revealed three matches with significant homology. An identity of approximately 67% was found between obcA and an ORF from Burkholderia ubonensis and about 53% identity was found between the obcA sequence and an ORF found in two related human pathogenic bacteria: Burkholderia pseudomallei and Burkholderia mallei.

Data were analysed using spss version 18 (SPSS Inc., Chicago, IL, USA). Proportions were compared using the χ2 test and ages were compared by means of a one-way analysis of variance (ANOVA). P-values of <0.05 were considered

statistically significant. The ethical committee of Hospital São João approved the study design in 2007. No specific consent was obtained from the patients as the data were used anonymously. As shown in Table 1, in the sample as a whole there were similar proportions of male and female patients. Patients followed in the southern area of the country represented 59% of the sample population. Dual infections (HIV-1 and HIV-2) accounted for a minority (3.6%) of cases. Around half of the patients were Portuguese citizens (213; 48.2%).

Guinea Bissau, buy RAD001 Cape Verde and Angola were the countries of origin of 33.5, 7.9 and 2.5% of the patients, respectively. The mode of transmission was mainly reported as heterosexual (260; 58.8%). Blood transfusions were the route for HIV-2 transmission in 15.4% of cases, but the proportion of cases attributed to blood transfusions has been declining over time. Injecting drug use was the mode of acquisition SAHA HDAC in 2.3% of patients and men who have sex with men accounted for 1.1%. Vertical transmission was rare (0.9%). The mode of transmission was not specified for 21.5% of the participants. The majority of the patients were asymptomatic at diagnosis (283; 64.0%). Lymphocyte CD4 cell count at diagnosis was available for 62% of the patients. Of these, 62 (22.6%) had a CD4 count <200 cells/μL. At the last follow-up evaluation, most patients remained treatment-naïve (200; 45.2%). However, 156 (35.3%) were on antiretroviral therapy, 14.5% of whom had experienced at least two different treatment regimens. During follow-up, at least 23.7% developed

AIDS. By the end of December 2007, 128 (29%) of the patients were alive; 82 (18.6%) had died. For 232 (52.5%), the outcome was unknown. HIV-2 infection diagnoses were distributed over time as follows: 1985 to 1989, 57 patients; 1990 to 1994, 83 patients; 1995 to 1999, 95 patients; 2000 to 2004, 127 patients and 2005 to 2007, 73 patients (Table 2). For seven patients, the year of diagnosis was not specified. Chlormezanone Before 1989, the majority of patients were male (39; 68.4%), had Portuguese nationality (45; 78.9%) and were living in the north of the country (44; 77.2%). The mean age at diagnosis was 31.0 (±14.7) and 37.8 (±8.9) years for male and female patients, respectively. Most patients were infected through heterosexual intercourse (31; 54.4%), but the proportion of HIV-2 infections attributed to blood transfusions was high (22; 38.6%). Forty-one individuals (71.9%) were asymptomatic at the time of diagnosis. From 1990 to 1994, the numbers of cases of newly diagnosed HIV-2 infection were nearly equal in men and women (41 men and 42 women). Heterosexual transmission remained the main transmission route (61.4%), followed by blood transfusion (31.3%).

No separated sample had a viral load > 200 copies/mL. Only two whole-blood

samples had a viral load of < 40 copies/mL compared with 19 of 21 separated samples (90%). All separated samples had an HIV-1 viral load of 54 copies/mL or less, i.e. nil had a significant viraemia (Fig. 1). The range of results for whole-blood samples was from ‘not detected’ to 3080 copies/mL; the mean was 629 copies/mL and the median 279 copies/mL. Further research in this important area is needed. HIV-1 RNA results that are above the cut-off in patients on treatment have much greater implications than a slightly inaccurate result in a patient off treatment. There is currently no evidence in the Compound C chemical structure literature which relates to the reproducibility of HIV RNA assays at low copy number relating to different periods of time pre-centrifugation in patients on ART. Therefore, until these data become available using current assays, including Roche TaqMan v2.0, we

suggest that plasma separation should occur at under 24 hours, ideally at under 8 hours. Close attention needs to be paid to the timing of plasma separation in patients on ART who are pregnant and enrolled in clinical trials. “
“The aim of this study was to develop a system for rapid and accurate real-time quantitative PCR (qPCR) identification and quantification of Botrytis cinerea, one of the major pathogens present on grapes. The intergenic spacer (IGS) region of the nuclear ribosomal DNA was used to specifically detect and quantify PLX4032 supplier B. cinerea. A

standard curve was established to quantify this fungus. The qPCR reaction was based on the simultaneous detection of OSBPL9 a specific IGS sequence and also contained an internal amplification control to compensate for variations in DNA extraction and the various compounds from grapes that inhibit PCR. In these conditions, the assay had high efficiency (97%), and the limit of detection was estimated to be 6.3 pg DNA (corresponding to 540 spores). Our method was applied to assess the effects of various treatment strategies against Botrytis in the vineyard. Our qPCR assay proved to be rapid, selective and sensitive and may be used to monitor Botrytis infection in vineyards. Many fungal and bacterial organisms, of which Botrytis cinerea is the most important, can infect grapes and cause a ‘bunch rot’ (Keller et al., 2003). The disease caused by B. cinerea, also known as ‘grey mould’, is arguably the most significant disease problem confronting the wine industry worldwide. The presence of grey mould on grapes is undesirable, as it lowers the quality of wines. Depending on the vintage, fungal infection rates can reach 15–25% of grapes, and wines prepared from infected grapes usually exhibit organoleptic defects, such as colour oxidation or the appearance of typical aromatic notes (‘moldy’, ‘rotten’), which are not appreciated by consumers (Cilindre et al., 2007).