Synergistic activity of gold nanoparticles with amphotericin B on persister cells of Candida tropicalis biofilms.

AIM: The antifungal activity was studied on sessile and persister cells (PCs) of Candida tropicalis biofilms of gold nanoparticles (AuNPs) stabilized with cetyltrimethylammonium bromide (CTAB-AuNPs) and those conjugated with cysteine, in combination with Amphotericin B (AmB). MATERIALS/METHODS: The PC model was used and synergistic activity was tested by the checkerboard assay. Biofilms were studied by crystal violet and scanning electron microscopy. RESULTS/CONCLUSIONS: After the combination of both AuNPs and AmB the biofilm biomass was reduced, with significant differences in architecture being observed with a reduced biofilm matrix. In addition, the CTAB-AuNPs-AmB combination significantly reduced PCs. Understanding how these AuNPs aid in the fight against biofilms and the development of new approaches to eradicate PCs has relevance for chronic infection treatment.

Interaction and antibacterial activity of ciprofloxacin with choline based ionic liquid and CTAB: A comparative spectroscopic study.

In this study, the complexation of potential chemo-therapeutic antibacterial drug, ciprofloxacin (CIP) with varying concentrations of surface active compounds (SACs) i.e., (N-(2-hydroxyethyl)-N,N-dimethyl-1-dodecanaminium bromide (12Cho.Br) and cetyltrimethylammonium bromide (CTAB) has been studied. Multispectroscopic techniques were exploited to carry out the study. The higher binding constant (Kb) value for CIP-CTAB than CIP-12Cho.Br obtained from fluorescence data revealed stronger binding of CTAB than 12Cho.Br, owing to the stronger hydrophobic-hydrophobic interaction betweeen CIP and CTAB compared to CIP and 12Cho.Br. The time resolve fluorescence decay shows changes in average lifetime (τavg) with the increasing concentration of 12Cho.Br and CTAB. The changes in τavg suggests that complex formation is taking place between CIP and 12Cho.Br / CTAB. Further, the formation of micelles by 12Cho.Br / CTAB and the effect of alkyl chain length was studied by dynamic light scattering (DLS) and zeta potential to confirm the drug complexation with 12Cho.Br and CTAB. The antibacterial activity has been performed for CIP and 12Cho.Br and CTAB. It was observed that in presence of lower concentrations of 12Cho.Br/ CTAB, the activity of the drug increased. The activity was also found cationic alkyl chain length dependent. Moreover, in-vitro cytotoxicity of CIP and its combinations with 12Cho.Br and CTAB was performed using MTT assay on HEK293 (Human embryonic kidney cells).

The antimicrobial and antiadhesion activities of micellar solutions of surfactin, CTAB and CPCl with terpinen-4-ol: applications to control oral pathogens.

The oral pathogen Streptococcus mutans is involved in tooth decay by a process that initiates with biofilm adhesion and caries development. The presence of other microbes such as Candida albicans may worsen the demineralization process. Since both microbes are virulent to the host and will proliferate under specific host immune deficiencies and systemic diseases, it is important to study antimicrobial substances and their effects on both pathogens. There are several antiseptic agents used to reduce plaque biofilm and its outcome (dental caries and/or periodontal disease). However, some of these have undesired effects. In the current study we investigated the antimicrobial and anti-adhesion properties of micellar solutions of surfactants and the plant natural product terpinen-4-ol (TP). The results revealed an increase in antimicrobial properties of the synthetic surfactants, cetylpyridinium chloride (CPC) and cetyltrimethylammonium bromide (CTAB), when mixed with TP. In addition, although surfactin, a biosurfactant, has little antimicrobial activity, it was demonstrated that it enhanced the effect of TP both as antimicrobial and anti-adhesion compound. Surfactin and the synthetic surfactants promote the antimicrobial activity of TP against S. mutans, the causal agent of tooth decay, suggesting specificity for membrane interactions that may be facilitated by surfactants. This is the first report on the successful use of surfactin in association with TP to inhibit the growth and adhesion of microbial pathogens. Surfactin has other beneficial properties besides being biodegradable, it has antiviral and anti-mycoplasma activities in addition to adjuvant properties and encapsulating capacity at low concentration.

Biofilm formation by Staphylococcus aureus and Salmonella spp. under mono and dual-species conditions and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite

Abstract The aim of this study was evaluated the biofilm formation by Staphylococcus aureus 4E and Salmonella spp. under mono and dual-species biofilms, onto stainless steel 316 (SS) and polypropylene B (PP), and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite. The biofilms were developed by immersion of the surfaces in TSB by 10 d at 37 °C. The results showed that in monospecies biofilms the type of surface not affected the cellular density (p > 0.05). However, in dual-species biofilms on PP the adhesion of Salmonella spp. was favored, 7.61 ± 0.13 Log10 CFU/cm2, compared with monospecies biofilms onto the same surface, 5.91 ± 0.44 Log10 CFU/cm2 (p < 0.05). The mono and dual-species biofilms were subjected to disinfection treatments; and the most effective disinfectant was peracetic acid (3500 ppm), reducing by more than 5 Log10 CFU/cm2, while the least effective was cetrimonium bromide. In addition, S. aureus 4E and Salmonella spp. were more resistant to the disinfectants in mono than in dual-species biofilms (p < 0.05). Therefore, the interspecies interactions between S. aureus 4E and Salmonella spp. had a negative effect on the antimicrobial resistance of each microorganism, compared with the monospecies biofilms.

Direct determination of surfactant effects on the uptake of gaseous parent and alkylated PAHs by crop leaf surfaces.

The partitioning of atmospheric polycyclic aromatic hydrocarbons (PAHs) into crop systems raises concerns about their potential harm to ecosystem and human health. To assess parent and alkylated PAHs accumulation accurately, the uptake of individual 7-isopropyl-1-methylphenanthrene (Retene), 3-methyl-phenanthrene (3-MP) and phenanthrene (Phe) by living maize, soybean and potato leaf surfaces, as well as the effects of cationic cetyltrimethylammonium bromide (CTMAB) and anionic sodium dodecyl benzene sulfonate (SDBS), were examined in situ using fiber-optic fluorimetry. For each of three PAH chemicals, the uptake achieved equilibrium between the air and living crop leaf surfaces within the 120-h monitoring period. There is inter-chemical and inter-species variability in terms of both the time required reaching equilibrium, the equilibrated adsorption concentration (EAC) and the overall air-surfaces mass transfer coefficient (kAS). The EAC of the three PAHs for each of the three crops' leaf surfaces increased with the number of alkyl substitutions on the aromatic ring. For any given PAHs, the EAC values followed the sequence of potato > soybean > maize, which was dominantly controlled by their leaf surface polarity index ((O+N)/C). The presence of CTMAB and SDBS increased the EAC of PAHs in the three crops' leaf surfaces by 6.5-17.1%, due to the plasticizing effect induced by the surface-sorbed surfactants, and the enhancement degree was closely associated with leaf-wax content and lg KOW values of PAHs. In addition, the two surfactants promoted the kAS values of the three chemicals by 7.7-23.3%. These results demonstrated that surfactants promoted the uptake of PAHs onto the crop leaf surfaces, potentially threatening the agricultural product safety.

Antimicrobial effectiveness of combinations of oxidant and chelating agents in infected dentine: an ex vivo confocal laser scanning microscopy study.

AIM: To evaluate the intratubular antimicrobial activity of several oxidant and chelating agents associated with or without surfactants in experimentally infected root canals, using confocal laser scanning microscopy. METHODOLOGY: Twenty-four dentine blocks from bovine incisors were contaminated for 5 days with Enterococcus faecalis (ATCC- 29212). Ten contaminated dentine specimens were irrigated for 5 min with 5.25% NaOCl followed by 17% EDTA for 2 min, and the other 10 with Hypoclean for 5 min followed by Tetraclean NA for 2 min. The remaining four specimens were used as positive and negative controls (two samples each). Then, dentine blocks were stained with Live/Dead BacLight for analysis of the remaining live or dead bacteria using confocal laser scanning microscopy. Comparison between and within groups was performed using the Mann-Whitney test for independent samples and the Wilcoxon signed-rank test, respectively. RESULTS: After exposure to irrigants, the positive control group had a median of 67.41% of viable bacteria (95% CI: 48.15, 78.9), whilst the NaOCl+EDTA group and Hypoclean+Tetraclean NA group had 3.77% (1.28, 15.92) and 0.87% (-0.42, 4.30) of viable bacteria, respectively. These results were significantly different from each other, both overall and distinct by region (cervical and medium third), or depth (superficial and deep layer) (P < 0.01 in all cases). CONCLUSIONS: The use of adjunctive agents reducing the surface tension associated with oxidant and chelating agents improved the antimicrobial activity of irrigating solutions and intratubular decontamination against Enterococcus faecalis, possibly due to better removal of the smear layer and deeper penetration into dentinal tubules.

Reduced Susceptibility to Antiseptics Is Conferred by Heterologous Housekeeping Genes.

Antimicrobial resistance is common in the microbial inhabitants of the human oral cavity. Antimicrobials are commonly encountered by oral microbes as they are present in our diet, both naturally and anthropogenically, and also used in oral healthcare products and amalgam fillings. We aimed to determine the presence of genes in the oral microbiome conferring reduced susceptibility to common antimicrobials. From an Escherichia coli library, 12,277 clones were screened and ten clones with reduced susceptibility to triclosan were identified. The genes responsible for this phenotype were identified as fabI, originating from a variety of different bacteria. The gene fabI encodes an enoyl-acyl carrier protein reductase (ENR), which is essential for fatty acid synthesis in bacteria. Triclosan binds to ENR, preventing fatty acid synthesis. By introducing the inserts containing fabI, ENR is likely overexpressed in E. coli, reducing the inhibitory effect of triclosan. Another clone was found to have reduced susceptibility to cetyltrimethylammonium bromide and cetylpyridinium chloride. This phenotype was conferred by a UDP-glucose 4-epimerase gene, galE, homologous to one from Veillonella parvula. The product of galE is involved in lipopolysaccharide production. Analysis of the E. coli host cell surface showed that the charge was more positive in the presence of galE, which likely reduces the binding of these positively charged antiseptics to the bacteria. This is the first time galE has been shown to confer resistance against quaternary ammonium compounds and represents a novel, epimerase-based, global cell adaptation, which confers resistance to cationic antimicrobials.

Biofilm formation by Staphylococcus aureus and Salmonella spp. under mono and dual-species conditions and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite.

The aim of this study was evaluated the biofilm formation by Staphylococcus aureus 4E and Salmonella spp. under mono and dual-species biofilms, onto stainless steel 316 (SS) and polypropylene B (PP), and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite. The biofilms were developed by immersion of the surfaces in TSB by 10 d at 37°C. The results showed that in monospecies biofilms the type of surface not affected the cellular density (p>0.05). However, in dual-species biofilms on PP the adhesion of Salmonella spp. was favored, 7.61±0.13Log10CFU/cm2, compared with monospecies biofilms onto the same surface, 5.91±0.44Log10CFU/cm2 (p<0.05). The mono and dual-species biofilms were subjected to disinfection treatments; and the most effective disinfectant was peracetic acid (3500ppm), reducing by more than 5Log10CFU/cm2, while the least effective was cetrimonium bromide. In addition, S. aureus 4E and Salmonella spp. were more resistant to the disinfectants in mono than in dual-species biofilms (p<0.05). Therefore, the interspecies interactions between S. aureus 4E and Salmonella spp. had a negative effect on the antimicrobial resistance of each microorganism, compared with the monospecies biofilms.

Antibiofilm Activity of Sodium Hypochlorite and Alkaline Tetrasodium EDTA Solutions.

INTRODUCTION: The aim of this study was to determine the antimicrobial activity of mixed alkaline tetrasodium EDTA (EDTANa4)/sodium hypochlorite (NaOCl) solutions with and without the addition of cetrimide (CTR) against Enterococcus faecalis biofilms. METHODS: The antimicrobial solutions were evaluated on a 3-week biofilm of E. faecalis grown on radicular dentin blocks. The irrigating solutions were 2.5% NaOCl group, 20% EDTANa4 group, 10% EDTANa4 group, 2.5% NaOCl/10% EDTANa4 group, 2.5% NaOCl/5% EDTANa4 group, 2.5% NaOCl/10% EDTANa4/0.2% CTR group, 2.5% NaOCl/5% EDTANa4/0.2% CTR group, and 0.9% saline solution group. Cell viability was determined by adenosine triphosphate assay, and culture techniques were used to determine colony-forming unit (CFU) counts. In groups with positive growth, the Pearson linear correlation coefficient r (-1 ≤ r ≤ 1) between relative light units (RLUs) and log10 (CFUs + 1) was calculated. The results of the bioluminescence adenosine triphosphate assay and CFUs of the groups were analyzed by means of analysis of variance and the Duncan test. RESULTS: There was Pearson correlation between RLUs and log10 (CFUs + 1). A similar reduction of RLUs in all NaOCl groups was seen, giving statistical differences with respect to the control. In the NaOCl groups there was no bacterial growth. The lowest antimicrobial efficacy was found for the EDTANa4 groups. CONCLUSIONS: Mixed alkaline EDTANa4/NaOCl solutions with and without the addition of CTR do not interfere with the antimicrobial activity of NaOCl.

PLGA-CTAB curcumin nanoparticles: Fabrication, characterization and molecular basis of anticancer activity in triple negative breast cancer cell lines (MDA-MB-231 cells).

Triple-negative breast cancers (TNBC) are aggressive cancers, which do not control by hormonal therapy or therapies that target HER-2 receptors. Curcumin (Cur) has shown cytotoxic effects in multiple cancer cell lines. However, its medical uses remain limited due to low aqueous solubility and poor bioavailability. Therefore, present study was aimed to fabricate the small positive charge curcumin nanoparticles (CN) by nanoprecipitation methods using PLGA and CTAB, and to evaluate its anticancer efficacy and underlying the mechanism in triple negative breast cancer cell lines (MDA-MB-231 cells). In in-vitro drug release assay, Cur was released from CN by flicking diffusion and anomalous transport process. CN showed a higher cellular incorporation than free Cur resulted in higher cytotoxicity. Checking the anticancer activity at the molecular level, Cur has shown to induce the reactive oxygen species production that subsequently causes the DNA damage and resulting in p38-MAPK activation. The p38-MAPK induce the expression of p16/INKK4a, p21/waf1/cip1 and p53 resulting in a reduction in the level of CDK2, CDK4, cyclin D1 and cyclin E and subsequently cell cycle arrest at G1/S and G2/M phase. It also reduces the expression of DNA repair gene, i.e. BRCA1, BRCA2, Rad51, Rad50, Mre11 and NBS1 resulting in apoptosis induction due to persistent DNA damage. This study presents an effective delivery of curcumin in TNBC cancer cells and it could open the new frontiers in clinical cancer chemotherapy.

Use of Plant Preservative Mixture™ for establishing in vitro cultures from field plants: Experience with papaya reveals several PPM™ tolerant endophytic bacteria.

KEY MESSAGE: Prevalence of diverse PPM™-tolerant endophytic bacteria in papaya, the broad-spectrum microbicide specified for use in plant tissue cultures, capable of surviving covertly in MS-based medium, with implications in contamination management. Plant Preservative Mixture™ was employed for establishing papaya (Carica papaya) tissue cultures from field explants. Comparing three recommended practices for controlling endogenous microbial contaminants, axillary shoot tips (1.0-1.5 cm) from cv. Arka Prabhath were treated with PPM™ 5% for 4 h (T1), 50% for 10 min (T2) or 100% for 10 min (T3) and cultured in MS-based papaya establishment medium (PEM). By 4-6 weeks, all treatments proved non-rewarding with cultures succumbing either to microbial contamination (80% in T1) or phytotoxicity effect/contamination (90% in T2 and 95% in T3). Another trial adopting a multi-step surface sterilization treatment (carbendazim-cetrimide-HgCl2) followed by culturing in 0.05% PPM-supplemented PEM showed 35% obvious bacterial contamination compared with 40% in control. Single colonies from pooled bacterial growths were tested on 0.1% PPM-incorporated nutrient agar (NA) registering 60% isolates as PPM sensitive. Twenty PPM-surviving isolates were selected and identified. This showed 85% Gram-positive bacteria including 80% under phylum Firmicutes (55% spore-forming Bacillaceae and 25% Staphylococcaceae) and 5% Actinobacteria, and 15% Gram-negative Proteobacteria. About 50% isolates remained wholly non-obvious upon culturing on PEM while the rest showed slow growth with many displaying growth enhancement upon host tissue extract supplementation. Culturing the isolates on PPM-supplemented NA indicated 90-95% as tolerating 0.05-0.1% PPM and 65% overriding 0.2% PPM. The isolates, however, did not display obvious growth in PPM-supplemented PEM where the spore formers survived. The results indicate the prevalence of diverse PPM™-tolerant endophytic bacteria in papaya most of which survive covertly in MS-based medium and the need for taking this into account while using PPM™ for contamination management.

Combination of selected enzymes with cetyltrimethylammonium bromide in biofilm inactivation, removal and regrowth.

Enzymes are considered an innovative and environmentally friendly approach for biofilm control due to their lytic and dispersal activities. In this study, four enzymes (ß-glucanase, α-amylase, lipase and protease) were tested separately and in combination with the quaternary ammonium compound cetyltrimethylammonium bromide (CTAB) to control flow-generated biofilms of Pseudomonas fluorescens. The four enzymes caused modest reduction of biofilm colony forming units (CFU). Protease, ß-glucanase and α-amylase also caused modest biofilm removal. CTAB combined with either ß-glucanase or α-amylase increased biofilm removal. Its combination with either ß-glucanase or protease increased CFU reduction. However, CTAB-protease combination was antagonist in biofilm removal. Long-term effects in biofilm mass reduction were observed after protease exposure. In contrast, biofilms treated with ß-glucanase were able to regrow significantly after exposure. Moreover, short-term respirometry tests with planktonic cells were performed to understand the effects of enzymes and their combination with CTAB on P. fluorescens viability. Protease and lipase demonstrated antimicrobial action, while α-amylase increased bacterial metabolic activity. The combination of CTAB with either protease or α-amylase was antagonistic, decreasing the antimicrobial action of CTAB. The overall results demonstrate a modest effect of the selected enzymes in biofilm control, either when applied alone or each one in combination with CTAB. Total biofilm removal or CFU reduction was not achieved and, in some cases, the use of enzymes antagonized the effects of CTAB. The results also propose that complementary tests, to characterize biofilm integrity and microbial viability, are required when someone is trying to assess the role of novel biocide - enzyme mixtures for effective biofilm control.

Adaptation to Biocides Cetrimide and Chlorhexidine in Bacteria from Organic Foods: Association with Tolerance to Other Antimicrobials and Physical Stresses.

Chlorhexidine (CH) and quaternary ammonium compounds (QAC), such as cetrimide (CE), are widely used as disinfectants because of their broad antimicrobial spectrum. However, their frequent use for disinfection in different settings may promote bacterial drug resistance against both biocides and clinically relevant antibiotics. This study analyzes the effects of stepwise exposure to cetrimide (CE) and chlorhexidine (CH) of bacteria from organic foods and previously classified as biocide-sensitive. Gradual exposure of these strains to biocides resulted in mainly transient decreased antimicrobial susceptibility to other antibiotics and to biocides. Biocide-adapted bacteria also exhibit alterations in physiological characteristics, mainly decreased heat tolerance, or gastric acid tolerance in CE-adapted strains, while bile resistance does not seem to be influenced by biocide adaptation. Results from this study suggest that changes in membrane fluidity may be the main mechanism responsible for the acquisition of stable tolerance to biocides.

Zwitterionic gold nanorods: low toxicity and high photothermal efficacy for cancer therapy.

Novel "zwitterionic" gold nanorods (Au NRs) were constructed through a facile ligand exchange process between cetyltrimethylammonium bromide (CTAB)-Au NRs and the zwitterionic block polymer {poly(2-methacryloyloxyethyl phosohorylcholine)-b-poly(lipoic methacrylate) (pMPC-b-pLA)}. In vitro, they exhibited low dark cytotoxicity and a high therapeutic efficacy to cancer cells. Their blood circulation half-life in vivo (t1/2, ∼10 h) was 20-fold longer than that of CTAB-Au NRs (t1/2, <30 min). After intravenous administration, they accumulated in tumour sites via an enhanced permeability and retention (EPR) effect and enabled destruction of human xenograft tumours in mice after exposure of the tumour location to NIR laser irradiation at 808 nm. These studies showed that the "zwitterionic" Au NRs had low toxicity and high photothermal efficacy both in vitro and in vivo due to the suprahydrophilic, biocompatible zwitterionic polymer coating layer. They may have the potential to be a promising NIR PTT agent in the biomedical field.

Antibacterial effect and physical properties of chitosan and chlorhexidine-cetrimide-modified glass ionomer cements.

AIMS: To compare antibacterial effect and physical properties of chitosan (CH) modified glass ionomer cement (GIC) (10% v/v), chlorhexidine-cetrimide (CHX-CT) modified GIC (2.5/2.5% w/w) and conventional GIC. MATERIALS AND METHODS: A total of fifty healthy children of age 7-12 years were selected and randomly assigned to class A and B for in vivo analysis. Slabs of CH modified GIC (Group II) along with slabs of conventional GIC (Group I, control) were cemented on buccal surfaces of maxillary molars (split-mouth technique) for class A children. Similarly, slabs of CHX-CT modified GIC (Group III) were cemented against control (Group I, control) in class B children. Slabs were assessed after 48 h for microbial load of Streptococcus mutans and Lactobacillus (LB) on mitis salivarius-bacitracin and Man Rogosa Sharpe agar media, respectively. Agar diffusion test was done to access the antibacterial effect of each group against Streptococcus muatns and LB. Slabs and cylinders of GICs were made for in vitro evaluation of compressive and flexure strength in each group. RESULTS: Comparison was done by nonparametric Kruskal-Wallis analysis followed by Dunn's multiple comparison test. Categorical groups were compared by Chi-square test. The increase in antibacterial activity (Group II > III > I) (P < 0.001) and marked increase in compressive and flexure strength (Group II > I > III) were observed. CONCLUSIONS: In the view of findings, it is concluded that CH modified GIC would be effective in inhibiting the bacteria associated with dental caries along with improved physical properties when compared with CHX-CT modified GIC and conventional GIC.

Polydimethyl siloxane based nanocomposites with antibiofilm properties for biomedical applications.

Polydimethyl siloxane (PDMS) is an excellent implant material for biomedical applications, but often fails as it is prone to microbial colonization which forms biofilms. In the present study CuO, CTAB capped CuO, and ZnO nanoparticles were tested as nanofillers to enhance the antibiofilm property of PDMS against Staphylococcus aureus and Escherichia coli. In general S. aurues (Gram positive and more hydrophobic) favor PDMS surface than glass while E. coli (Gram negative and more hydrophilic) behaves in a reverse way. Incorporation of nanofillers renders the PDMS surface antibacterial and reduces the attachment of both bacteria. These surfaces are also not cytotoxic nor show any cell damage. Contact angle of the material and the cell surface hydrophobicity influenced the extent of bacterial attachment. Cell viability in biofilms was dependent on the antimicrobial property of the nanoparticles incorporated in the PDMS matrix. Simple regression relationships were able to predict the bacterial attachment and number of dead cells on these nanocomposites. Among the nanocomposites tested, PDMS incorporated with CTAB (cetyl trimethylammonium bromide)-capped CuO appears to be the best antibacterial material with good cyto-compatibility. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1075-1082, 2017.

Efficacy of antimicrobial solutions against polymicrobial root canal biofilm.

AIM: To evaluate the antibiofilm activity of 2.5% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), 2% alexidine (ALX) and 0.2% cetrimide (CTR) alone and in combination on mature polymicrobial root canal biofilms on human dentine using confocal laser scanning microscopy (CLSM). METHODOLOGY: Twenty-eight human dentine specimens were infected for 21 days with microbial samples collected from infected root canals of three volunteers. Antibiofilm activity of the irrigating solutions was evaluated after 3 min of contact time under CLSM. For quantification purposes, bioimage_L software was used. The variables evaluated were the log10 of total biovolume (µm3 ) and percentage of live cells (green) population. Statistical analysis of both variables was performed using an anova test and a post hoc Duncan test to determine significant clusters amongst groups. The variable green population percentage was previously subjected to the normalized Anscombe transformation. RESULTS: The NaOCl group had a total biovolume and percentage of live cells significantly lower than the other groups (P Ë‚ 0.001). The addition of 0.2% CTR significantly increased the antimicrobial effect of 2% CHX (P Ë‚ 0.001). There were no significant differences between 0.2% CTR, 2% ALX and the combination of both (P Ë‚ 0.05). CONCLUSIONS: Overall, 2.5% NaOCl dissolved and killed bacteria significantly more efficiently when used against polymicrobial mature biofilm on human dentine. Cetrimide improved the antimicrobial activity of chlorhexidine and alexidine.

Biocide tolerance, phenotypic and molecular response of lactic acid bacteria isolated from naturally-fermented Aloreña table to different physico-chemical stresses.

Lactic acid bacteria (LAB) isolated throughout the fermentation process of Aloreña table olives were found to be resistant at least to three antibiotics (Casado Muñoz et al., 2014); however, most were sensitive to the biocides tested in this study (with minimum inhibitory concentrations [MIC] below the epidemiological cut-off values). 2-15% of the isolates were found to be biocide resistant: Leuconostoc Pseudomesenteroides, which were resistant to hexachlorophene, and Lactobacillus pentosus to cetrimide and hexadecylpiridinium. We analyzed the effect of different physico-chemical stresses, including antimicrobials, on the phenotypic and genotypic responses of LAB, providing new insights on how they become resistant in a changing environment. Results indicated that similar phenotypic responses were obtained under three stress conditions: antimicrobials, chemicals and UV light. Susceptibility patterns to antibiotics changed: increasing MICs for ampicillin, chloramphenicol, ciprofloxacin, teicoplanin and tetracycline, and decreasing the MICs for clindamycin, erythromycin, streptomycin and trimethoprim in most strains. Statistically, cross resistance between different antibiotics was detected in all stress conditions. However, expression profiles of selected genes involved in stress/resistance response (rpsL, recA, uvrB and srtA) differed depending on the stress parameter, LAB species and strain, and the target gene. We conclude that, despite the uniform phenotypic response to stresses, the repertoire of induced and repressed genes differs. So, a search for a target to improve stress tolerance of LAB, especially those of importance as starter/protective cultures or probiotics, may depend on the individual screening of each strain, even though we could predict the antibiotic phenotypic response to all stresses.

Hybrid catechin silica nanoparticle influence on Cu(II) toxicity and morphological lesions in primary neuronal cells.

Morphological alterations compromising inter-neuronal connectivity may be directly linked to learning-memory deficits in Central Nervous System neurodegenerative processes. Cu(II)-mediated oxidative stress plays a pivotal role in regulating redox reactions generating reactive oxygen species (ROS) and reactive nitrogen species (RNS), known contributors to Alzheimer's disease (AD) pathology. The antioxidant properties of flavonoid catechin have been well-documented in neurodegenerative processes. However, the impact that catechin encapsulation in nanoparticles may have on neuronal survival and morphological lesions has been poorly demonstrated. To investigate potential effects of nano-encapsulated catechin on neuronal survival and morphological aberrations in primary rat hippocampal neurons, poly(ethyleneglycol) (PEG) and cetyltrimethylammonium bromide (CTAB)-modified silica nanoparticles were synthesized. Catechin was loaded on silica nanoparticles in a concentration-dependent fashion, and release studies were carried out. Further physicochemical characterization of the new nano-materials included elemental analysis, particle size, z-potential, FT-IR, Brunauer-Emmett-Teller (BET), thermogravimetric (TGA), and scanning electron microscopy (SEM) analysis in order to optimize material composition linked to the delivery of loaded catechin in the hippocampal cellular milieu. The findings reveal that, under Cu(II)-induced oxidative stress, the loading ability of the PEGylated/CTAB silica nanoparticles was concentration-dependent, based on their catechin release profile. The overall bio-activity profile of the new hybrid nanoparticles a) denoted their enhanced protective activity against oxidative stress and hippocampal cell survival compared to previously reported quercetin, b) revealed that morphological lesions affecting neuronal integrity can be counterbalanced at high copper concentrations, and c) warrants in-depth perusal of molecular events underlying neuronal function and degeneration, collectively linked to preventive nanotechnology in neurodegeneration.

In Vitro Synergism between Azithromycin or Terbinafine and Topical Antimicrobial Agents against Pythium insidiosum.

We describe here in vitro activity for the combination of azithromycin or terbinafine and benzalkonium, cetrimide, cetylpyridinium, mupirocin, triclosan, or potassium permanganate. With the exception of potassium permanganate, the remaining antimicrobial drugs were active and had an MIC90 between 2 and 32 µg∕ml. The greatest synergism was observed for the combination of terbinafine and cetrimide (71.4%). In vivo experimental evaluations will clarify the potential of these drugs for the topical treatment of lesions caused by Pythium insidiosum.