, including cholesterol-lowering agents (monacolins), an antihypertensive substance (γ-aminobutyric acid) and an antioxidant (dimerumic acid) (Aniya et al., 2000; Lin et al., 2008; Pattanagul et al., 2008). However, the problem of safety emerged in 1995 when Blanc et al. (1995a) identified monascidin A, an antibacterial compound in RFR, as a nephrotoxic metabolite, citrinin. Thus, control of the production of citrinin is essential to increase the safety of Monascus-related products and extend their applications. In the past

decade, researchers have made considerable progress towards improving Monascus-related products using a process of optimization and traditional mutation breeding methods (Wang et al., 2004; Chen & Hu, 2005; Sayyad et al., 2007). Recently, some biosynthetic gene clusters involved in the biosynthesis of secondary metabolites of Monascus spp., such as citrinin learn more and

selleck products monacolin K, have been identified (Shimizu et al., 2007; Chen et al., 2008b). Based on the genetic information, a genetic modification method has also been proposed (Fu et al., 2007; Jia et al., 2010). Secondary metabolite production is controlled at an upper hierarchical level by many global mechanisms, in which many proteins encoded by genes not linked to the biosynthetic gene clusters are also involved in modulating fungal secondary metabolism, such as transcription factor, histone deacetylase, DNA methyltransferase, signalling proteins such as MAP kinases and cAMP-dependent protein kinase (Fox & Howlett, 2008). Heterotrimeric G-proteins, acting within G-protein signalling pathways to regulate multiple

physiological processes and that generally respond to environmental cues such as pH, temperature and nutrition, are also found to be involved in the regulation of secondary metabolite production in some toxigenic fungi (Hicks et al., 1997; Seo & Yu, 2006; Yu et al., 2008). Heterotrimeric G-proteins consist of three subunits: Gα, Gβ and Gγ. They function as ‘molecular switches’ in G-protein signalling Pregnenolone pathways to regulate the duration and intensity of the signal, eventually going on to regulate downstream cell processes. Most characterized filamentous fungi possess three Gα proteins belonging to three distinct groups, Groups I, II and III, of which Group I is the most extensively studied (Li et al., 2007). Accumulating evidence has suggested that individual Group I Gα protein regulates multiple pathways. For example, dominant activating mutations in fadA in Aspergillus nidulans blocked both sterigmatocystin production and asexual sporulation, and the deletion of GzGPA1 in Gibberella zeae resulted in female sterility and enhanced deoxynivalenol and zearalenone production (Hicks et al., 1997; Yu et al., 2008).

7%) and 38 were female (25.3%). The mean age of the group was 44.3 ± 8.3 years, and the median age was 44 years. The

mean age of the male patients was 44.8 ± 8.2 years and that of the female patients was 43 ± 8.8 years; this difference was not significant. Of the 150 patients, selleck 19 (17.0%) of the male patients and five (13.2%) of the female patients were >50 years old. Men were more often single than of other marital status, and women were more often married or widowed (P<0.001). The most common type of cohabitation was living with partner or children, or both (48.7%); cohabitation was significantly more frequent in women than in men (65.8%vs. 42.9%, respectively; P=0.015). A summary of the sociodemographic, epidemiological and clinical data is presented in Table 1. The mean PHS value was 52.3 ± 8.8 and the mean MHS value was 49.3 ± 9.9. The distribution of mean values for the MOS-HIV questionnaire is shown in Table 2. We found that women had lower scores than men in Pain (P=0.038) and Cognitive Functioning (P=0.037), with no differences in the other HRQL domains. Patients >50 years old had higher scores than the youngest age category in Pain patients without children got higher scores than patients with children in Smad inhibitor (P=0.018), Health Distress (P=0.018) and Cognitive

Functioning (P=0.004). Single patients (P=0.020), those who lived alone (P=0.006) and those without children (P<0.001) had higher scores for General Forskolin clinical trial Health Perceptions. In this last group, patients without children got higher scores than patients with children in Energy (P=0.018), Quality of Life (P=0.007), PHS (P=0.005) and MHS (P=0.012) scores were higher in patients with children. We found no significant differences for educational background or income level. Former smokers had higher scores than other patients in the Health Distress domain (P=0.032). Patients with a history of injecting drug use (IDU) had lower

scores than other patients in General Health Perceptions (P=0.059), Pain (P=0.005), Physical Functioning (P=0.003), Social Functioning (P=0.070) and PHS (P<0.001). Patients who stated that they were homosexual had lower scores than other patients in General Health Perceptions (P=0.007), Pain (P=0.034), Physical Functioning (P=0.002) and MHS (P<0.001). Furthermore, patients who contracted HIV infection through sharing of needles among heterosexual injecting drug users had lower scores than other patients in General Health Perceptions (P=0.034). In terms of immune system status, we did not find a relationship between the domains of the MOS-HIV questionnaire and the variables CD4 cell count and viral load. However, patients with CDC category stage C disease (European classification) had higher scores than other patients in Mental Health (P=0.023), Energy (P=0.050), Cognitive Functioning (P=0.046), Quality of Life (P=0.018) and MHS (P=0.025).

Reactivation of latent virus is common in those with advanced immunosuppression and frequently does not cause end-organ disease. Detection of CMV in urine, blood or BAL without evidence of end-organ involvement implies CMV infection but not disease. CMV isolation in BAL (by culture or PCR)

is common in individuals with HIV infection, who have low CD4 T-cell counts [120,121]. Typical symptoms are dry non-productive cough and exertional dyspnoea with fever; and this presentation is similar to many other pulmonary conditions [120,122]. Hypoxaemia is often marked [120]. The chest radiograph and CT scan most often show bilateral interstitial infiltrates or ground glass attenuation, but unilateral alveolar consolidation, bilateral nodular opacities, find protocol pleural effusions or rarely cavities or hilar adenopathy may occur [120,122,123]. There may be concomitant evidence of extra-pulmonary CMV [120] and a dilated eye examination should be performed to rule out CMV retinitis. The major diagnostic challenge is to differentiate CMV shedding in respiratory secretions from cases with CMV pneumonitis. Culture, positive PCR or antigen

assay for CMV from BAL or biopsy specimen do not distinguish CMV shedding from pneumonitis, and hence must be interpreted with caution [124,125]. A negative culture result, with its high negative predictive value, however, does reasonably exclude CMV pneumonia [126]. Diagnosis of CMV pneumonia requires a biopsy specimen to provide evidence of pulmonary learn more involvement in association with a compatible clinical syndrome (category III recommendation). Evidence Ureohydrolase of intranuclear or intracytoplasmic viral inclusions, positive immunostaining for CMV antigens or detection of CMV by molecular techniques such as in situ hybridization on a pulmonary biopsy specimen establishes the diagnosis in the setting of a compatible clinical syndrome

[120]. CMV replication in the respiratory tract is most frequently only a marker of immunosuppression and not of pneumonia. The majority of individuals in whom microbiological tests on BAL, or biopsy, demonstrate CMV should not receive treatment for CMV (category III recommendation). This approach is supported by evidence that when treatment is withheld, in individuals with evidence of CMV on BAL or biopsy, clinical outcome is not adversely altered [127]. However, the benefits of treatment to the select subset of individuals who have evidence of a compatible clinical syndrome, positive microbiology and histology for CMV and no alternative diagnosis have been suggested by retrospective case series that show improved clinical outcomes with treatment [121]. The management of individuals with positive histology for CMV but identification of a second pulmonary pathogen is also controversial.

Reactivation of latent virus is common in those with advanced immunosuppression and frequently does not cause end-organ disease. Detection of CMV in urine, blood or BAL without evidence of end-organ involvement implies CMV infection but not disease. CMV isolation in BAL (by culture or PCR)

is common in individuals with HIV infection, who have low CD4 T-cell counts [120,121]. Typical symptoms are dry non-productive cough and exertional dyspnoea with fever; and this presentation is similar to many other pulmonary conditions [120,122]. Hypoxaemia is often marked [120]. The chest radiograph and CT scan most often show bilateral interstitial infiltrates or ground glass attenuation, but unilateral alveolar consolidation, bilateral nodular opacities, SCH772984 pleural effusions or rarely cavities or hilar adenopathy may occur [120,122,123]. There may be concomitant evidence of extra-pulmonary CMV [120] and a dilated eye examination should be performed to rule out CMV retinitis. The major diagnostic challenge is to differentiate CMV shedding in respiratory secretions from cases with CMV pneumonitis. Culture, positive PCR or antigen

assay for CMV from BAL or biopsy specimen do not distinguish CMV shedding from pneumonitis, and hence must be interpreted with caution [124,125]. A negative culture result, with its high negative predictive value, however, does reasonably exclude CMV pneumonia [126]. Diagnosis of CMV pneumonia requires a biopsy specimen to provide evidence of pulmonary Alvelestat cell line involvement in association with a compatible clinical syndrome (category III recommendation). Evidence Bcl-w of intranuclear or intracytoplasmic viral inclusions, positive immunostaining for CMV antigens or detection of CMV by molecular techniques such as in situ hybridization on a pulmonary biopsy specimen establishes the diagnosis in the setting of a compatible clinical syndrome

[120]. CMV replication in the respiratory tract is most frequently only a marker of immunosuppression and not of pneumonia. The majority of individuals in whom microbiological tests on BAL, or biopsy, demonstrate CMV should not receive treatment for CMV (category III recommendation). This approach is supported by evidence that when treatment is withheld, in individuals with evidence of CMV on BAL or biopsy, clinical outcome is not adversely altered [127]. However, the benefits of treatment to the select subset of individuals who have evidence of a compatible clinical syndrome, positive microbiology and histology for CMV and no alternative diagnosis have been suggested by retrospective case series that show improved clinical outcomes with treatment [121]. The management of individuals with positive histology for CMV but identification of a second pulmonary pathogen is also controversial.

[44] Light-weight, titanium-impregnated nylon and cotton fabrics will offer the greatest comfort and sun protection in hot and humid regions and can be layered in cooler and dryer regions. Ruxolitinib order Washing clothing with photoprotective laundering agents, such as Rit Sun Guard, will offer photoprotection through one’s favorite clothes at low cost. Besides responsible selection of sun protective clothing, the consumer-traveler should be a responsible wearer of photoprotective clothing by avoiding wet and tightly fitted clothing and gaps of uncovered skin at the ankles, wrists, waist, and neck between the shirt collar

and hat. In addition to wide-brimmed hats and photoprotective clothing, sunglasses also provide photoprotection for the skin and, most importantly, the eyes and eyelids, by preventing the development of several ocular disorders including periorbital skin cancers, cataracts, pterygia, photokeratitis, snow blindness, and possibly retinal melanomas and age-related macular degeneration.[48, 49] There is no world standard UV protection rating system for sunglasses. The first national standard rating system for UV protection for sunglasses was introduced by Australia in 1971. The existing national standard UV protection rating systems for sunglasses are compared in Table 4. Travelers should choose the highest AG-014699 mouse UV protection-rated sunglasses as indicated on the required hangtags. Sunglass UV protection depends on several factors

including shape and fit, and lens color and UV-filtering and reflecting abilities.[48, PRKACG 49] Sunglass lenses should fit close to the face, not touch the eyelashes,

hug the temples, and merge into broad temple arms or straps. Darker lenses do not necessarily filter more UV light and can trigger pupillary dilation which allows unfiltered wavelengths of UV and visible-spectrum blue light (400–440 nm) to reach the retina.[50] Chronic retinal exposure to visible-spectrum blue light in the wavelength range of 400 to 440 nm is a risk factor for age-related macular degeneration.[50-53] The color of sunglass lenses can influence contrast, color vision, and depth and width perception.[50-53] Orange and yellow lenses provide the best protection from both UV and visible blue light, with blue and purple lenses providing insufficient protection.[50-53] The effects of sunglass lens colors on visual perception are compared in Table 5.[50-53] A variety of special use sunglasses are recommended for travelers engaging in active water sports, such as body-boarding, jet-skiing, kite-boarding, wake-boarding, wind sailing, and water skiing. Water sunglasses (goggles) have air vents to prevent fogging and increased buoyancy to prevent sinking if lost. Glacier sunglasses (goggles) provide more UV filtration and reflection and are recommended for travelers engaging in winter and high altitude sports, such as cross-country skiing, downhill skiing, snowboarding, glacier hiking, and mountain climbing.

The resulting plasmid (pKX23) was verified by nucleotide sequencing and used as the template plasmid to make the linear recombineering substrate (Fig. 3b). The http://www.selleckchem.com/products/Roscovitine.html linear DNA was used to recombineer in RSW358 strains with pJAK12, pJAK14, or pJAK16. Selection of the recombinants was for Gmr. Transformants numbered > 4000 mL−1 for each,

and as expected, all were white on X-Gal-IPTG medium. Recombinants of pJAK12, pJAK14, and pJAK16 [pKX32 (Spr Gmr), pKX34 (Kmr Gmr), and pKX36 (Cmr Gmr), respectively] were verified by nucleotide sequencing. The aacC1-encoding SalI fragment was removed from each plasmid by digestion with SalI, religation, and transformation. Spr, Kmr, or Cmr transformants were selected, as appropriate for FG-4592 pJAK12, pJAK14, and pJAK16, respectively. As expected, Spr/Kmr/Cmr Gms transformants were blue on X-Gal-IPTG medium. Nucleotide sequencing confirmed the structures. The recombinants were named pJAK12 Blue, pJAK14 Blue, and pJAK16 Blue (Fig. 2b).

We also used the method to construct an oriTIncP-Gmr cassette and to provide the recombineering substrate for targeting it to the cat gene of pSIM9 (Fig. 2c). The template plasmid was constructed from pCR2.1 TOPO using HindIII, BamHI, NotI, XhoI, and XbaI (Fig. 3c). The oriT-encoding PCR product, made from pAA56 with flanking BamHI and NotI recognition sites (Table 2c), was inserted at the TA-cloning site, oriented by PCR with the appropriate primers, and verified by nucleotide sequencing to give pKR1. The aacC1 gene was cloned into NotI- and

XhoI-cleaved pKR1 to give the oriT-Gmr plasmid pKR2. HRI (260 bp) was cloned into HindIII- and BamHI-cleaved pKR2 to give pKR6, and HRII (275 bp) was ligated to XhoI- and XbaI-cleaved pKR6 to give pKR7. The MCS region of pKR7, which should have the elements for the recombineering substrate, was verified by nucleotide sequencing. Plasmid pKR7 was then used to make the recombineering substrate by PCR using primers Urease P1 and P8 (Table 2c). Gmr selection led to > 4000 colonies mL−1. One typical Gmr Cms oriTIncP pSIM9 derivative (pKR8) was shown by nucleotide sequencing to have the expected structure. In summary, we developed a method for making recombineering substrates with PCR primers that can be ≤ 35 nucleotides long (the ‘short-primer’ method). The method uses restriction endonuclease–based molecular cloning techniques to link GEs and regions of homology to make a recombineering substrate. A downside of the short-primer method is that it takes somewhat longer than the long-primer method to obtain the desired recombinant (about twice as long if the substrate is made from three cloned segments). The HRs of the short-primer method are easily changed to target the GEs to a different site. In addition, cloning of the segments requires that the PCR primers work to give the desired fragment, and each intermediate plasmid can be verified.

, 2005, 2008). Mature miRNA present in synaptoneurosome fractions of adult brain tissue derives at least in part from processing of local precursors (Lugli et al., 2008). Evidence suggests that NMDAR activation selleck chemical results in the proteolytic liberation of Dicer from the postsynaptic density and

subsequent activation of its RNAase III activity (Lugli et al., 2005). Mature miRNAs regulated by this mechanism should be rapidly elevated at synaptic sites following NMDAR activation and LTP induction, while the corresponding precursor levels should decrease. Such a pattern of regulation was not observed in the present study, but our measurements of miRNA expression in whole dentate gyrus homogenates (rather than synaptic fractions) cannot be used to address the question of local precursor processing in LTP. Selleck GSK3235025 Taken together, however, these studies have revealed an unexpected regulation of the mature miRNA expression by NMDAR-dependent and transcription-independent mechanisms. Coordinate action of many miRNAs is crucial for biological processes,

such as cell fate determination and apoptosis. Co-transcriptional regulation is one way by which such coordination is thought to be achieved (Cheng et al., 2007; He et al., 2007). Evidence suggests that miR-132 and -212 are co-transcriptionally regulated from a stable intron of a cryptic non-coding RNA (Vo et al., 2005). An important common target of miR-132 and -212 is the Rac/Rho-family p250GAP. In cultured hippocampal neurons, activity-dependent GNE-0877 expression of miR-132 regulates dendritic morphogenesis by decreasing synthesis of p250GAP (Vo et al., 2005; Wayman et al., 2008). The transcription of miR-132 in this context is CREB dependent, and stimulated by NMDAR and brain-derived neurotrophic factor (BDNF)-activated

signaling pathways. In vitro studies also indicate a key role for miR-132 transcription in the homeostatic regulation of MeCP2 during neural development (Klein et al., 2007). The present work on LTP in the adult gyrus shows that transcription of pri-miR-132 and -212 is strongly dependent on mGluR rather than NMDAR activation. Changes in mature miR-132 and miR-212 during LTP reflected the opposing effects of mGluR and NMDAR signaling. Interestingly, while net levels of mature miRNA were significantly increased, no changes in the expression of p250GAP or MeCP2 protein were detected. Taken together, the results suggest that transcriptional regulation of miR-132/212 and its impact on target protein expression differs substantially between the developmental setting of embryonic neurons and LTP in the adult brain. The present study gives the first insights into regulation of miRNA expression during LTP in the adult mammalian brain.

The PCR product was analyzed in a 2% agarose Ixazomib concentration gel and purified from the gel using the gel extraction kit (Qiagen). The purified fragment was then inserted into the cloning vector (pGEMT; Promega) to confirm their identity. Plasmid isolation and purification were done using the Wizard plus SV Minipreps DNA purification

system (Promega). The presence of insert in the plasmid was checked by double digestion with restriction enzymes NotI plus NcoI. Plasmid containing the insert was sequenced using an automatic DNA Sequencer (310 Genetic Analyser; Applied Biosystems, Foster City, CA). The catR promoters (Pcat300, Pcat924) were then inserted into the promoter-less xylanase/pAN56-1 plasmid to check their functionality. Pcat300 and Pcat924 were re-amplified using the above-mentioned primers and Pfu DNA polymerase to get blunt-ended amplified products. Promoter-less xylanase/pAN56-1 vector was digested with EcoRV and de-phosphorylated. Digested and de-phosphorylated vector was ligated to Pfu-amplified Pcat300 and Pcat924 promoter fragments. Both ligated mixtures were click here electroporated in JM110-competent cells using gene pulser (Bio-Rad). The plasmids were isolated with Qiagen’s spin column according

to the instructions of the manufacturer. The presence of insert in the plasmids and orientation of the Pcat300 and Pcat924 in promoter-less xylanase/pAN-56-1 was checked by digestion with NcoI. Transformation of A. niger by constructs (Pcat300/xylanase/pAN56-1, Pcat924/xylanase/pAN56-1) was carried out by electroporation as described by Sanchez & Aguirre (1996). Transformed spores were spread on minimal medium agar plates containing 175 μg mL−1 hygromycin (Biogene; Imperial Biomedics) as the selective agent, and incubated at 37 °C (Tilburn et al., 1983; Malardier et al., 1989). Transformants were observed after 36–48 h at 37 °C. Individual clones were transferred to fresh Sabouraud’s/hygromycin plates. selleck Genomic DNA of putative transformants was extracted and amplified by the E. coli ori primers (Varadarajalu & Punekar, 2005) to confirm that each construct had

been integrated into the genome of A. niger. The transformants were further evaluated quantitatively for xylanase production by growing in seed medium under shaking conditions (200 rpm) for 48 h at 28 °C (inoculum size was 2 × 106 spores per flask) and then 10% inoculum was transferred in wet wheat bran (production medium pH 6.0) under static conditions for 96 h. The AlX enzyme from production medium was extracted by shaking at 30 °C for 2 h using 0.05 M phosphate buffer (pH 8.0) and filtered through a wet muslin cloth by squeezing. The extract was centrifuged at 6000 g for 5 min. Clear supernatant sample from each transformant was taken and used for the enzyme assay. Xylanase activity was estimated by quantifying the release of reducing sugar and expressed in terms of IU mL−1 (Gupta et al., 2000).

harveyi (Fig. 4), which encodes a V. cholerae

QS pathway (Hammer & Bassler, 2008). As with V. cholerae, the maximal transformation frequency occurred with the WT V. harveyi strain, which produces both CAI-1 and AI-2. Transformation decreases when only CAI-1 or AI-2 was provided, and was most impaired in the absence of either autoinducer (Fig. 4). We also measured transformation frequency of V. cholerae autoinducer-deficient recipient in response to WT V. parahaemolyticus and V. fischeri autoinducer donors. Transformation efficiency of these Vibrio strains followed a pattern of comEA-lux expression that matched the corresponding donor strains; the V. parahaemolyticus buy FK228 strain used produces both CAI-1 and AI-2 and promoted transformation with IWR 1 a frequency similar to V. harveyi. The V. fischeri strain tested (and another sequenced V. fischeri strain, data not shown) only encode for luxS (and not cqsA), and thus produce AI-2, but not CAI-1. Vibrio fischeri poorly promoted DNA uptake by the V. cholerae recipient (Fig. 4), consistent with AI-2 playing a minor role in natural transformation. Taken together, these observations support a model that

V. cholerae can switch to the competent state and acquire DNA horizontally in a chitinous environmental biofilm by responding to autoinducer signals derived from members of the multispecies consortium. Induction of the competence program in V. cholerae requires the chitin-responsive

TfoX pathway and the autoinducer-responsive QS pathway. When both systems are functional, DNA uptake machinery facilitates the transport of extracellular DNA into the bacterial cell, where it may be incorporated into the genome by homologous recombination (Hamilton & Dillard, 2006). Many Vibrios encode for chitin utilization and O-methylated flavonoid competence genes (Hunt et al., 2008; Gulig et al., 2009; Ng & Bassler, 2009; Pollack-Berti et al., 2010), which suggests the possibility that natural transformation may be a conserved mechanism for both pathogenic and nonpathogenic Vibrios to horizontally acquire virulence and other genes within a community. Recognizing that many Vibrios possess V. cholerae-like QS circuits and produce CAI-1 and AI-2, we examined the relationship between autoinducers production and DNA uptake. Specifically, we showed that (1) V. cholerae efficiently activated a comEA-lux reporter in response to self-produced autoinducers as well as purified autoinducers and (2) a V. cholerae autoinducer-deficient strain readily acquires DNA when co-cultured with purified autoinducers and also with autoinducers produced by other Vibrios within a chitinous mixed-species biofilm. These results support a model that V. cholerae can switch to the competent state in a chitinous environmental biofilm by responding to autoinducer molecules derived from members of the multispecies consortium.

, 2003; Zhao et al., 2003). We then discuss AI-2 production pathways and the implications of AI-2 production in oomycte cross-kingdom communication. Two morphological and phylogenetically distinct Phytophthora species, and a species from the closely related genus Pythium, were used in this study. Phytophthora nicotianae (Syn. P. MK-8669 ic50 parasitica) isolate 1B11, Phytophthora sojae (genotype I) isolate 23G8, and Pythium aphanidermatum isolate 18H1 were maintained in clarified 20% vegetable juice medium supplemented with

1.5% agar (CV8A) at 23 °C. ZFF was prepared from nutrient-depleted zoospore suspensions at high densities. A 5-mm2 CV8A mycelial plug was seeded in 10% CV8 in 90-mm Petri dishes. The dishes were incubated at 23 °C in the dark for 3 days for P. sojae, 4 days for P. aphanidermatum, and 1–2 weeks Abiraterone for P. nicotianae to induce sporangia. After the seed plugs and medium were removed, the mycelial mats were rinsed five times with sterile-distilled water (SDW) to eliminate nutrients from

the remaining medium. The drained mycelial mats were incubated for 16–18 h for P. sojae and P. aphanidermatum, and 1 week for P. nicotianae under fluorescent light at 23 °C. When numerous sporangia formed, the mats were rinsed an additional five times with SDW to remove residues from the medium. The dilution factor for the 10% CV8 was then 1.08 × 109 as measured experimentally. To induce zoospore release, the mats were flooded with 8 mL of chilled SDW and kept under light until the desired zoospore density was reached. (-)-p-Bromotetramisole Oxalate The density for 1B11 was up to 106 zoospores mL−1 in 1 h; for 23G8 and 18H1, it was up to 5 × 104 and 3 × 104 zoospores mL−1 in 3 h, respectively. All procedures were performed under sterile conditions to prevent bacterial contamination. To obtain ZFF, zoospore suspensions were filtered through a sterile miracloth to remove mycelia, sporangia, and other structures,

and then vortexed briefly to facilitate chemical release. The suspensions were then filtered through a 0.2-μm syringe filter to remove the cysts. ZFF was used fresh or stored at −20 °C. The bacterial AI-2 reporter Vibrio harveyi BB170 [luxN∷TnS] (ATCC BAA-1117) was used to test the activity of ZFF and detect the presence of AI-2. The assay was conducted using a combined protocol based on the procedures described previously (Bassler et al., 1997; DeKeersmaecker & Vanderleyden, 2003). Briefly, BB170 was cultured overnight in MB medium and then diluted 10 000 × into AB medium. Aliquots of 90 μL from the resulting overnight culture were dispensed into each well of a 96-well plate, followed by the addition (10 μL per well) of test solutions. The plate was then incubated at 30 °C with aeration. Light production was monitored using a CCD camera after 3 h of incubation for a period of 8 h, and the integrated optical density (IOD) was measured using labworks image acquisition and analysis software (UVP, CA).