coli O157:H7, compost samples were individually treated with antimicrobials that targeted gram-positive bacteria (crystal violet), gram-negative bacteria (streptomycin) and eukaryotic species (amphotericin B and cycloheximide) (Fig. 2). The survival of E. coli O157:H7 improved significantly in the presence of the eukaryotic inhibitor cycloheximide and the CFU mL−1 remained relatively constant throughout

the incubation period. No significant differences were observed in the reduction of E. coli O157:H7 compared with the control in the presence of crystal violet, amphotericin B or streptomycin. Statistical comparisons between the slope means between each treatment and the control yielded P=0.578 (crystal violet), P=0.258 (streptomycin), P=0.993 (amphotericin B) and P=0.002 (cycloheximide). These data suggest that cycloheximide-sensitive eukaryotic species were primarily

responsible for the observed decline of E. coli Ensartinib research buy click here O157:H7. DGGE analysis was used to monitor the shifts in populations in compost samples (Fig. 3). At 25 °C, the banding patterns of fungal species remained similar between cycloheximide-treated and -untreated samples, except for some bands that repeatedly appeared on day 8 and day 10 of the cycloheximide-treated samples. In contrast, a dramatic shift in protist populations was observed at 25 °C between untreated and treated samples (Fig. 3). Notably, none of the prominent bands seen in control compost samples from days 4 to 12 were observed in the lanes for the cycloheximide-treated samples. Protist clone libraries were created from cycloheximide-treated and -untreated compost samples that were incubated at 25 °C.

This set of samples was chosen for further analysis because DGGE results suggested that protists play a more significant role in E. coli O157:H7 decline in our compost model than the fungal species. The numbers of OTUs (observed richness) present in the control library FER at day 0 were 19 compared with 17 OTUs in the cycloheximide-treated library (Table 1). The Chao1 estimator (predicted richness) estimated that the two day 0 libraries had 28 and 35 OTUs, respectively. Therefore, we estimate that 68% and 49% of the species present in the control and the cycloheximide-treated samples at day 0 were covered by the clone libraries. libshuff was used to determine the differences between the clone libraries at each time point. Pairwise comparisons between the two day 0 libraries generated P values of 0.58 and 0.67, suggesting that the two libraries are statistically similar (Fig. 4). Similar analyses were performed on days 6, 8 and 12 samples and all control libraries were statistically different from the corresponding cycloheximide-treated sample libraries (Fig. 4). From the blast analysis, days 0, 8 and 12 control libraries were largely composed of the ciliophora Arcuospathidium cultriforme (Day 0) and Onychodromopsis flexilis (Days 8 and 12).

96 (95% CI 0.94–0.99) per 1 year older; P = 0.009] and IDU [ARR 1.38 (95% CI 0.98–1.94)] vs. sexually transmitted HIV [ARR 1.38 (95% CI 0.98–1.94); P = 0.01] were independent predictors of abortion. To evaluate the impact of HIV-related factors on the incidence of first abortion, we then focused on the 60 events that occurred during 4349 PYFU after HIV diagnosis [crude incidence rate 13.8 per 1000 PYFU (95% CI 10.7–17.8)]. We observed a high incidence rate of induced abortion among women who acquired HIV by IDU [23.0 per 1000 PYFU (95% CI 15.5–34.0)] and those who were not on cART and were aware of being HIV-infected before pregnancy

[7.6 per 1000 PYFU (95% CI 19.5–39.9)]. Further, women who self-reported a fear of vertical HIV transmission [22.9 per 1000 PYFU (95% CI 15.3–34.2)] or of con-natal malformations [19.7 per 1000 PYFU (95% CI 10.7–35.1)] had high abortion incidence rates. Conversely, a low

incidence rate was observed in women who were already Selleckchem Doxorubicin aware of their HIV infection and who were on cART at the time of the abortion [8.6 per 1000 PYFU (95% CI 5.7–12.8)] and those who declared a monthly income higher than €800 [9.4 per 1000 PYFU (95% CI 6.1–14.4)]. The abortion incidence rate in migrant women living with HIV was even lower [3.5 per 1000 PYFU (95% CI 0.5–24.8)]. In the multivariable model, the risk of first abortion was significantly lower in more recent calendar years. In fact, compared with the period before 1990, the risk of first BTK inhibitors abortion was 0.47 (95% CI 0.22–0.99; P = 0.04) in 1990–1999 and 0.37 (95% CI 0.13–1.02; P = 0.05) in 2000–2010. Among women who were aware of their HIV infection before pregnancy, the current use of cART was protective against abortion [receiving vs. not receiving cART, ARR 0.54 (95% CI 0.28–1.04); P = .06]; women who had a diagnosis at pregnancy did not show an increased risk of abortion compared with those who were already aware of their infection and who were off

cART [HIV diagnosed during pregnancy vs. already aware of HIV Cediranib (AZD2171) infection and not receiving cART, ARR 0.84 (95% CI 0.37–1.90); P = 0.68]. Fear of vertical transmission was strongly associated with abortion after HIV diagnosis: women who were concerned about infecting the child showed a twofold higher risk of abortion compared with those who were not [ARR 1.90 (95% CI 1.02–3.56); P = 0.04]. Monthly income lower than €800 [ARR 1.76 (95% CI 0.99–3.11); P = 0.05 vs. monthly income ≥ €800] and younger age [per 1 year older, ARR 0.95 (95% CI 0.91–1.00); P = 0.05] were also found to be independent predictors of first abortion after HIV diagnosis. The risk of abortion in women who became sexually active before 15 years of age tended to be higher [ARR 1.65 (95% CI 0.91–2.98); P = 0.09].

Cytokeratin 5 and its partner cytokeratin 14 form dimers that help give tissue its integrity. Without the presence of these Cabozantinib ic50 cytokeratins, tissue becomes fragile and small injuries can cause tissue to fall apart and blisters to form. These cytokeratins have also been shown to be enhanced in hyperproliferative situations such

as wound healing [33, 39]. These data suggest that ZDV treatment impairs the ability of oral tissue to heal itself. In this study, ZDV treatment induced the expression of cytokeratin 10, particularly at the 6-, 12- and 24-h time-points (Figs 5 and 8). Increased levels of cytokeratin 10 in drug-treated gingival epithelium may be an attempt by the tissue to protect itself against damage caused by ZDV [31, 40, 41]. Additionally, it has been shown that cytokeratin 10 is more strongly expressed in both oral lesions and hyperproliferative epidermis compared

with ordinary epidermis [42]. Thus, the elevated levels of cytokeratin 10 may be linked to the proliferative effect of ZDV on treated rafts. Additionally, the normal balance of cytokeratin proliferation and differentiation may be disrupted upon injury and under pathological conditions [43-45]. Involucrin expression is induced by the same pathway as cytokeratin 5. In addition to a change in cytokeratin expression, envelope formation is a hallmark of terminal differentiation. In order for the envelope to be formed correctly, the envelope precursors and transglutaminase, the enzyme responsible for the assembly of the envelope, must be expressed HIF inhibitor not at the correct time and level during the differentiation process [37]. Involucrin is a component of the cornified envelope. Involucrin is specifically expressed in the suprabasal layers of the epidermis [37], while in the spinous and granular layer, involucrin accumulates as a non-cross-linked precursor. During the final stages of keratinocyte differentiation, involucrin becomes cross-linked to other proteins to form the cornified envelope [37]. Involucrin expression, like that of cytokeratin 5, is regulated by the specificity protein (Sp1) [37], and in our study the expression

of involucrin, like that of cytokeratin 5, was decreased in response to ZDV treatment. A lack of involucrin available for cross-linking may explain the lack of a vaculated, cornified layer seen in ZDV-treated tissues and may account for the fragility of oral tissues in patients on HAART. Induction of cytokeratin 6 expression in protease inhibitor-treated rafts [26, 27], as well as a slight increase in cytokeratin 10 expression in ZDV-treated tissues, suggested the possibility that HAART drugs, including ZDV, were causing damage to the gingival epithelium. To examine this possibility, we looked at the expression patterns of cytokeratin 6, a wound-healing keratin which is activated in response to injury in the suprabasal layer of stratified epithelium.

Cytokeratin 5 and its partner cytokeratin 14 form dimers that help give tissue its integrity. Without the presence of these KU-57788 cell line cytokeratins, tissue becomes fragile and small injuries can cause tissue to fall apart and blisters to form. These cytokeratins have also been shown to be enhanced in hyperproliferative situations such

as wound healing [33, 39]. These data suggest that ZDV treatment impairs the ability of oral tissue to heal itself. In this study, ZDV treatment induced the expression of cytokeratin 10, particularly at the 6-, 12- and 24-h time-points (Figs 5 and 8). Increased levels of cytokeratin 10 in drug-treated gingival epithelium may be an attempt by the tissue to protect itself against damage caused by ZDV [31, 40, 41]. Additionally, it has been shown that cytokeratin 10 is more strongly expressed in both oral lesions and hyperproliferative epidermis compared

with ordinary epidermis [42]. Thus, the elevated levels of cytokeratin 10 may be linked to the proliferative effect of ZDV on treated rafts. Additionally, the normal balance of cytokeratin proliferation and differentiation may be disrupted upon injury and under pathological conditions [43-45]. Involucrin expression is induced by the same pathway as cytokeratin 5. In addition to a change in cytokeratin expression, envelope formation is a hallmark of terminal differentiation. In order for the envelope to be formed correctly, the envelope precursors and transglutaminase, the enzyme responsible for the assembly of the envelope, must be expressed LY294002 Racecadotril at the correct time and level during the differentiation process [37]. Involucrin is a component of the cornified envelope. Involucrin is specifically expressed in the suprabasal layers of the epidermis [37], while in the spinous and granular layer, involucrin accumulates as a non-cross-linked precursor. During the final stages of keratinocyte differentiation, involucrin becomes cross-linked to other proteins to form the cornified envelope [37]. Involucrin expression, like that of cytokeratin 5, is regulated by the specificity protein (Sp1) [37], and in our study the expression

of involucrin, like that of cytokeratin 5, was decreased in response to ZDV treatment. A lack of involucrin available for cross-linking may explain the lack of a vaculated, cornified layer seen in ZDV-treated tissues and may account for the fragility of oral tissues in patients on HAART. Induction of cytokeratin 6 expression in protease inhibitor-treated rafts [26, 27], as well as a slight increase in cytokeratin 10 expression in ZDV-treated tissues, suggested the possibility that HAART drugs, including ZDV, were causing damage to the gingival epithelium. To examine this possibility, we looked at the expression patterns of cytokeratin 6, a wound-healing keratin which is activated in response to injury in the suprabasal layer of stratified epithelium.

The difference between our current and our previous studies suggests state-dependency in the form of a task-dependent role for PMd during online performance and offline consolidation

of implicit sequence-specific learning of a visuomotor task. Given its anatomical location and functional connectivity, the PMd is a likely convergence CP-673451 solubility dmso point for cognition and motor control. PMd is generally associated with explicit declarative aspects of motor learning. While the PMd has been implicated in facilitating the transition between implicitly learned movements that constitute a sequence (Mushiake et al., 1991), activity in the PMd is reduced when explicit awareness of the implicit motor sequences is gained (Hazeltine et al., 1997; Honda et al., 1998). During online learning it is likely that the PMd serves to enhance implicit sequence-specific learning by linking specific movements which are dependent upon sensory cues (Nowak et al., 2009; Taubert et al., 2010). This role may be particularly important during interleaved practice and may explain why anodal transcranial direct current stimulation over the PMd during constant repetitive Ibrutinib manufacturer practice does not result in improved consolidation of performance gains (Nitsche

et al., 2003; Kantak et al., 2012). In contrast, early offline consolidation of information relating to sequencing of action selection may interfere with early consolidation of more procedural elements relating to individual ADAMTS5 movements, which are most likely represented in M1

(Muellbacher et al., 2002; Wilkinson et al., 2010). This may result from early offline consolidation of information being more reliant on a declarative memory and thus more explicit. This assumption is consistent with observations of differential rates of consolidation for explicit declarative memories relative to procedural memory (Brown & Robertson, 2007a; Ghilardi et al., 2009; Galea et al., 2010) and competition between procedural and declarative memory systems (Brown & Robertson, 2007a,b; Galea et al., 2010). Interference may occur even when explicit instruction is not given and participants have not autogenously acquired declarative knowledge of a sequence through practice (Vidoni & Boyd, 2007). Therefore, reducing the cortical excitability of PMd through 1 Hz rTMS during early offline consolidation may relieve suppression of procedural representations in M1 during this critical period and facilitate an early boost in procedural learning not seen at later stages of offline consolidation (Hotermans et al., 2008). Another interesting result was the lack of dissociation between implicit motor sequence learning for the 5 Hz and sham stimulation groups. Relative to the sham control group, one might expect the 5 Hz group to show the opposite effect to that induced by 1 Hz rTMS.

For each experimental session a new word list was presented. The list was composed of complexity-matched words (see Supporting Information). During the mental activity, subjects were instructed to imagine the movements from a first person perspective and to employ kinesthetic cues (e.g. the feeling of the pen in their hand). The anodal tDCS was administered for 13 min during the whole course of the MP. Continuous direct currents were transferred by saline-soaked surface sponge electrodes (surface 20 cm2) and delivered by a clinical microcurrent stimulator (Soterix, USA) with a maximum output of 2 mA. Five different electrode montages

were tested to find the optimal position for DC stimulation in increasing the neuroplastic effects of mental imagery on motor

performance. The excitatory tDCS was applied over the: (i) right selleck compound M1, (ii) right premotor area (PMA), (iii) right SMA, (iv) right cerebellar hemisphere, and (v) left dorsolateral prefrontal cortex. For M1 tDCS, the anode electrode was positioned above C3 (international 10-20 system) (Nitsche et al., 2003b). For stimulation of the premotor cortex, it was moved 2 cm forward and 2 cm to the midline relative to the M1 position (Nitsche et al., 2003b). The SMA tDCS was performed with the anode electrode placed 2 cm anterior to the vertex (position Cz), in the sagittal midline (Cunnington et al., 1996). For DC stimulation of the dorsolateral prefrontal cortex, the anode electrode was positioned 5 cm forward relative to C3 (Nitsche et al., 2003b). In all cases, the reference electrode was placed above the contralateral orbit. For cerebellar tDCS, electrodes were placed with selleck screening library one (anode electrode) over the right cerebellar hemisphere, 3 cm lateral to the inion (Ugawa et al., 1995), and the other over the deltoid muscle (Ferrucci et al., 2008). These methods of electrode montage have been used in previous studies and been shown to be effective in the modulation of cerebral activity. The order of stimulation condition was counterbalanced across subjects. The anodal tDCS was administered with a current strength of 2 mA. In

the sham session, tDCS was applied over the M1 for 30 s, Beta adrenergic receptor kinase a method shown to achieve a good level of blinding (Gandiga et al., 2006). In each experimental session, motor performance was assessed by the handwriting test. This test measured legibility and writing time, important elements in handwriting performance (Bonney, 1992). Handwriting is a complex perceptual–motor skill that includes fine motor control (hand manipulation, bilateral integration, and motor planning) (Feder & Majnemer, 2007). For the test, the subjects were instructed to copy a six-word set with the non-dominant hand on a blank sheet of paper positioned on a table to the left of the subject. The word list was presented approximately three inches away from the paper. The handwriting task was performed with spontaneous production, free from the influence of the writing instructions.

For each experimental session a new word list was presented. The list was composed of complexity-matched words (see Supporting Information). During the mental activity, subjects were instructed to imagine the movements from a first person perspective and to employ kinesthetic cues (e.g. the feeling of the pen in their hand). The anodal tDCS was administered for 13 min during the whole course of the MP. Continuous direct currents were transferred by saline-soaked surface sponge electrodes (surface 20 cm2) and delivered by a clinical microcurrent stimulator (Soterix, USA) with a maximum output of 2 mA. Five different electrode montages

were tested to find the optimal position for DC stimulation in increasing the neuroplastic effects of mental imagery on motor

performance. The excitatory tDCS was applied over the: (i) right JQ1 order M1, (ii) right premotor area (PMA), (iii) right SMA, (iv) right cerebellar hemisphere, and (v) left dorsolateral prefrontal cortex. For M1 tDCS, the anode electrode was positioned above C3 (international 10-20 system) (Nitsche et al., 2003b). For stimulation of the premotor cortex, it was moved 2 cm forward and 2 cm to the midline relative to the M1 position (Nitsche et al., 2003b). The SMA tDCS was performed with the anode electrode placed 2 cm anterior to the vertex (position Cz), in the sagittal midline (Cunnington et al., 1996). For DC stimulation of the dorsolateral prefrontal cortex, the anode electrode was positioned 5 cm forward relative to C3 (Nitsche et al., 2003b). In all cases, the reference electrode was placed above the contralateral orbit. For cerebellar tDCS, electrodes were placed with LY294002 clinical trial one (anode electrode) over the right cerebellar hemisphere, 3 cm lateral to the inion (Ugawa et al., 1995), and the other over the deltoid muscle (Ferrucci et al., 2008). These methods of electrode montage have been used in previous studies and been shown to be effective in the modulation of cerebral activity. The order of stimulation condition was counterbalanced across subjects. The anodal tDCS was administered with a current strength of 2 mA. In

the sham session, tDCS was applied over the M1 for 30 s, 17-DMAG (Alvespimycin) HCl a method shown to achieve a good level of blinding (Gandiga et al., 2006). In each experimental session, motor performance was assessed by the handwriting test. This test measured legibility and writing time, important elements in handwriting performance (Bonney, 1992). Handwriting is a complex perceptual–motor skill that includes fine motor control (hand manipulation, bilateral integration, and motor planning) (Feder & Majnemer, 2007). For the test, the subjects were instructed to copy a six-word set with the non-dominant hand on a blank sheet of paper positioned on a table to the left of the subject. The word list was presented approximately three inches away from the paper. The handwriting task was performed with spontaneous production, free from the influence of the writing instructions.

The robustness of the trees was estimated by posterior probabilities. The nucleotide sequences reported in this paper have been PFT�� datasheet submitted to GenBank (FJ798929–FJ798951; GU256228–GU256245). The abundances of picoplanktonic cyanobacteria and heterotrophic bacteria were different in the lake basins in early June, 2008. In Northern Baikal picoplanktonic

cyanobacteria of the genera Synechococcus, Cyanobium and Synechocystis developed in huge numbers. They were dominated by an endemic Baikalian autotrophic picoplankton species –Synechocystis limnetica, which constituted 20% of the total picocyanobacterial number at depths of 0–25 m. As a whole, the numbers of picocyanobacteria reached 268 000 cells mL−1 at a depth of 5 m; the abundance of heterotrophic bacteria was about 288 000 cells mL−1 in the upper 25-m layer (Fig. 1). Thus, the share of picocyanobacteria in the total bacterial plankton number was about 50%, in biomass – 68%. At this time, the development of autotrophic picoplankton in Southern Baikal was low, and the numbers of picocyanobacteria were 12 400 cells mL−1 in the 0–25-m layer (Fig. 1). The main components of picocyanobacteria communities were species of Synechococcus

and Cyanobium genera, but, in contrast to the Northern basin, the contribution of S. limnetica to the total abundance did not exceed 4%. The abundance of bacteria in the Southern basin was high and averaged 1 780 000 mL−1 BCKDHA in the 0–25-m layer Palbociclib solubility dmso (Fig. 1). The share of the picocyanobacteria in total bacterial plankton abundance was only 1%,

in biomass – 3%. PCR products were obtained from both Northern and Southern Baikal water samples: each sample exhibited five bands that approximately ranged from 350 to 500 bp. All five bands of g23 amplicons from Northern Baikal water samples and only three bands from Southern Baikal were successfully reamplified. We constructed clone libraries of the purified g23 gene PCR products obtained from two stations. The recovery efficiency of g23 gene fragments from Southern Baikal was lower and only 70% of the clones contained correct g23 inserts within this clone library. In total, 23 clones from Northern Baikal and 18 from Southern Baikal were sequenced and translated (g23 amino acid sequence from 118 to 289 in the coliphage T4 sequence, Parker et al., 1984). The predicted amino acid sequences from Lake Baikal were variable in length from 105 to 143 residues. Each clone was designated as N0508 (Northern Baikal clone library) or S0508 (Southern Baikal), followed by band and clone numbers. The most similar based on blast hits were the g23 clones from marine, paddy fields and T4 cyanophages (from 70% and higher). The highest identity was observed between S0508/2-4 clone and CS26 marine clone (89%) (Fig. 2). Two highly conserved amino acid motifs of g23 marine sequences uncovered by Filée et al.

The material that was

retained inside this membrane (fraction SF-SK10-100R, 45 mg) was eluted on HPSEC as a single LEE011 research buy peak (Fig. 2c), with Mw 41 kDa and dn/dc=0.148. It was composed of glucose (51%), galactose (28%) and mannose (21%) and its 13C NMR spectrum (Fig. 3b) contained many C-1 signals, indicating the presence of a complex heteropolysaccharide. Methylation analysis (Table 2) indicated that all galactosyl units were present as nonreducing end-units (Galp and Galf), together with Glcp units. The Manp units were mainly 6-O-substituted, with small amounts of 2,6-di-O-substituted residues, while the Glcp units were 2-O-, 4-O-, 2,3-di-O-, 4,6-di-O- and 2,6-di-O-substituted residues. Substitution at HO-6 of the Manp and Glcp units was also shown by DEPT (Fig. 3b, inset), which provided inverted signals at δ 68.7 and 69.0. In its 13C

NMR spectrum, C-1 signals at δ105.1 and 105.6 corresponded to the nonreducing end-units of β-Galf. The signals at δ 102.6, 102.9 and 103.0 probably arose from C-1 of β-Glcp units. The anomeric configuration of these units was confirmed by their low-field C-1 resonances and also by their 1JC−1, H−1 of 161.5, CAL-101 concentration 164.2 and 160.0 Hz. The remaining C-1 signals at δ 100.7 and 100.2 belong to the α-pyranose series, due to their high-field C-1 resonances and 1JC−1, H−1 (174.4 and 171.5 Hz, respectively) (Agrawal, 1992; Duus et al., 2000; Bubb, 2003). The signals of O-substituted C-2, C-3 and C-4 could be seen at δ 87.5 (C-3), δ 84.9 and 83.3 (C-2) and δ 81.5 (C-4). The material that passed over this membrane (fraction SF-SK10-100E, 66 mg) had high glucose content (79%), with small amounts of mannose (10%) and galactose (11%), indicating the presence of a

glucan. This fraction still had Selleckchem Lumacaftor a heterogeneous elution profile on gel permeation (Fig. 2c) and due to its small amount was not further purified. However, its 13C NMR spectrum (Fig. 3c) showed β-configurations, due to low-field C-1 signals at δ 103.7 and 103.0. Moreover, it is possible to observe (13)- and (16)-linked Glcp units, due to the presence of a signal at δ 86.2, characteristic of O-substituted C-3, and to the presence of inverted signals at δ 68.8 and 69.0 in the DEPT experiment (Stuelp et al., 1999; Carbonero et al., 2001; Cordeiro et al., 2003). Thus, this glucan resembles a lentinan-type β-glucan. A similar glucan was isolated from the lichen Thamnolia vermicularis var. subuliformis by Olafsdottir et al. (2003) and had a backbone of β-d-(13)-linked glucopyranosyl units branched with a single β-d-(16)-linked unit for every third unit of the backbone. In an attempt to find the isolichenan, we also analyzed the fraction SW, which was obtained in low yield. This fraction was composed of galactose (60.0%), mannose (22.5%) and glucose (17.5%).

aureus controls its biofilm so as to discover novel compounds capable of this website inhibiting or dispersing biofilms without allowing bacteria to develop drug resistance. The diverse mechanisms that have been reported for biofilm control in S. aureus include quorum sensing, protease, DNase, cis-2-decenoic acid, d-amino acids, phenol-soluble polypeptides, several

surface proteins, and pH change (Boles & Horswill, 2011). Particularly, the activation of agr quorum-sensing and protease treatment in S. aureus inhibited its own biofilm formation and dispersed the established biofilms (Vuong et al., 2000; Boles & Horswill, 2008). A serine Esp protease in Staphylococcus epidermidis inhibited S. aureus biofilm formation and nasal colonization (Iwase et al., 2010). However, the target of these agr controlled protease and the specific

target of Esp protease is not known (Boles & Horswill, 2008; Iwase et al., 2010). Recently, we have shown that various Actinomycetes strains produce a large see more amount of protease that rapidly dispersed S. aureus biofilm (Park et al., 2012). In the present study, more diverse bacteria are used to screen for S. aureus biofilm reduction. We find that two Pseudomonas aeruginosa supernatants dispersed S. aureus biofilm and contained high protease activities. Another study goal is to identify a main antibiofilm component and a possible mechanism of protease-involved biofilm dispersal. Transcriptional analysis and phenotypic assays are conducted to confirm that S. aureus Methamphetamine triggers its biofilm dispersal through the accelerated effect of protease activity. All experiments were conducted at 37 °C, and Luria-Bertani (LB) medium was used for culturing all strains (Table 1). Two S. aureus strains (ATCC 25923 and ATCC 6538) were obtained from the Korean Agricultural Culture Collection and used to reinforce our findings in two different strains. To identify a main antibiofilm protease, thirteen P. aeruginosa PAO1 transposon

mutants (Jacobs et al., 2003) were obtained from the University of Washington Genome Center (Supporting information, Table S1). To obtain culture supernatants, a fresh single colony of bacteria was inoculated and cultured in LB at 250 r.p.m. for 24 h. All bacterial supernatants were filtered with a 0.45 μm filter to completely remove any bacteria before further use. Fresh culture supernatants were used in every experiment. Crystal violet, casamino acid, succinic acid, calcium carbonate, sodium phosphate, ethanol, soluble starch, and potassium phosphate were of analytical grade. DNase I (Cat. 79254) was purchased from Qiagen (Valencia, CA), and RNase A (Cat. 12091021) was purchased from Life Technologies (Grand Island, NY). For the cell growth measurements, the optical density was measured at 600 nm using a spectrophotometer (UV-160, Shimadzu, Japan). Each experiment was performed with at least two independent cultures.