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PubMed2006

Bacteriostatic polymer film immobilization.

Coatings of quaternary ammonium tertiary structures (QUATS) copolymerized with 4-vinylbenzylthymine (VBT) exhibited high antimicrobial activity against Escherichia coli. Immobilization of QUATS improves environmental performance by preventing release of antibacterials to the environment, helping to preclude the emergence of resistant strains. The crosslinking immobilization scheme reported herein provides a more environmentally benign and more inexpensive synthesis than previously reported, thus reducing the use of solvents, energy, and production time. Development of water soluble, thymine-based photopolymers was inspired by the UV-induced 2pi + 2pi photocyclodimerization of thymine in DNA. Copolymers of 4-vinylbenzylthymine and trimethylammonium chloride, triethylammonium chloride, or dimethyloctylammonium chloride were synthesized in different monomer ratios. The antibacterial properties were tested by coating VBT:QUATS in sterilized petri dishes, crosslinking under short UV light, spraying with aqueous suspensions of bacterial cells, air drying, and then applying agar media to promote bacterial growth. The plates were incubated for 24 h at 37 degrees C. The number of viable cells ranged from 17 to 0% growth. Immobilized VBT:QUAT copolymers are antiseptic surfaces that can be produced in an environmentally benign fashion.

Anti-Bacterial Agents↗
PubMed2006

Antifungal and antibacterial properties of a silver-loaded cellulosic fiber.

The skin is the interface between the body and the environment. Each skin type has a specific skin physiology and is more or less adapted for protection against multiple stress factors. Textiles on the other hand are the tissues with the longest contact to the human skin. They play a critical role especially in skin conditions with an increased rate of bacterial and fungal infections like atopic dermatitis and hyperhidrosis, and in diabetic patients and aged skin. The present study demonstrates the antifungal and antibacterial effects of SeaCell Active in an in vitro test system against Candida albicans (DSM 11225), Candida tropicalis (ATCC 1169), and Candida krusei (ATCC 6258). Furthermore, the antibacterial activity of fibers with different amounts of SeaCell Active fibers in a dose-dependent manner against Staphylococcus aureus (ATCC 22923) and Escherichia coli (ATCC 35218) could be demonstrated. If this fiber seems to be suited for bioactive textiles in specific anatomical regions and skin conditions with a susceptibility for fungal and bacterial infections due to Candida species, namely Staphylococcus aureus and Escherichia coli, must be examined by means of further investigations, especially in vivo tests in human, considering allergic and toxic effects of the fiber.

Anti-Bacterial Agents↗
PubMed2006

A lectin with antifungal and mitogenic activities from red cluster pepper (Capsicum frutescens) seeds.

A monomeric mannose/glucose-binding lectin, with a molecular mass of 29.5 kDa and an N-terminal sequence GQRELKL showing resemblance to that of the lectin-like oxidized low-density lipoprotein receptor from the rabbit, has been isolated from the seeds of red cluster pepper Capsium frutescens L. var. fasciculatum. The protocol involved anion exchange chromatography on diethylamino ethanol-cellulose and Q-Sepharose and fast protein liquid chromatography on Mono Q. Its hemagglutinating activity toward rabbit erythrocytes was inhibited by D: -mannose and glucose, specifically. The activity was stable from 0 to 40 degrees C, reached a maximum at pH 7 and 8, and was potentiated by Ca2+ and Mn2+ ions. The lectin showed strong mitogenic activity toward spleen cells isolated from BALB/c mice. The mitogenic activity, which reached a peak at a lectin concentration of 0.27 microM, was inhibited specifically by D(+)-mannose. The lectin was capable of inhibiting the germination of Aspergillus flavus and Fusarium moniliforme spores and hyphal growth in the two fungi.

Animals↗
PubMed2020

Natural brominated phenoxyphenols kill persistent and biofilm-incorporated cells of MRSA and other pathogenic bacteria.

Due to a high unresponsiveness to chemotherapy, biofilm formation is an important medical problem that frequently occurs during infection with many bacterial pathogens. In this study, the marine sponge-derived natural compounds 4,6-dibromo-2-(2',4'-dibromophenoxy)phenol and 3,4,6-tribromo-2-(2',4'-dibromophenoxy)phenol were found to exhibit broad antibacterial activity against medically relevant gram-positive and gram-negative pathogens. The compounds were not only bactericidal against both replicating and stationary phase-persistent planktonic cells of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa; they also killed biofilm-incorporated cells of both species while not affecting biofilm structural integrity. Moreover, these compounds were active against carbapenemase-producing Enterobacter sp. This simultaneous activity of compounds against different growth forms of both gram-positive and gram-negative bacteria is rare. Genome sequencing of spontaneous resistant mutants and proteome analysis suggest that resistance is mediated by downregulation of the bacterial EIIBC phosphotransferase components scrA and mtlA in MRSA likely leading to a lower uptake of the molecules. Due to their only moderate cytotoxicity against human cell lines, phenoxyphenols provide an interesting new scaffold for development of antimicrobial agents with activity against planktonic cells, persisters and biofilm-incoporated cells of ESKAPE pathogens. KEY POINTS: • Brominated phenoxyphenols kill actively replicating and biofilm-incorporated bacteria. • Phosphotransferase systems mediate uptake of brominated phenoxyphenols. • Downregulation of phosphotransferase systems mediate resistance.

Animals↗
PubMed2006

Antibacterial effect of two mineral trioxide aggregate (MTA) preparations against Enterococcus faecalis and Streptococcus sanguis in vitro.

The antibacterial effects of gray-colored MTA (GMTA) and white-colored MTA (WMTA) against Enterococcus faecalis and Streptococcus sanguis were assessed in vitro using the tube dilution test. Broth tubes were prepared and divided into experimental and control groups. Aliquots of each of the tested microorganisms were taken from a stock culture and added to each experimental and positive control group. All groups were incubated at 37 degrees C and evaluated for turbidity at 0, 1, 24, 48, and 72-hour time periods. A direct correlation was found between GMTA and WMTA concentrations and their antibacterial effect. Tubes containing GMTA in concentrations of 50, 25, and 12.5 mg/ml did not show E. faecalis growth at any of the time periods tested whereas tubes containing WMTA showed E. faecalis growth at all concentrations and time periods tested. Statistically significant differences were found between tubes containing GMTA in concentrations of 50, 25 and 12.5 mg/ml and tubes containing similar concentrations of WMTA (p < 0.001). Tubes containing GMTA in concentrations of 50, 25, 12.5, 6.25, and 3.12 mg/ml and tubes containing WMTA in concentrations of 50, 25, and 12.5 mg/ml did not show S. sanguis growth at any of the time periods tested. Statistically significant differences were found between tubes containing GMTA in concentrations 6.25 and 3.12 mg/ml and tubes containing similar concentrations of WMTA (p < 0.001). It appears that the susceptibility of E. faecalis and S. sanguis to MTA differed and that GMTA requires lower concentrations than WMTA to exert the same antibacterial effect against each of the microorganisms tested.

Aluminum Compounds↗
PubMed2005

Insertional mutation of marA vitiates inducible multiple antimicrobial resistance in Salmonella enterica subsp. enterica serovar Choleraesuis.

marA has been shown to mediate a multiple antimicrobial resistance (MAR) phenotype following induction in some members of the Enterobacteriaceae. When Salmonella Choleraesuis was exposed to inducing agents they displayed higher minimal inhibitory concentrations (MIC) to multiple antimicrobial agents and an increase in marA expression as determined by northern hybridization analysis. The objective of the present study was to determine if mutation of marA vitiated multiple antimicrobial resistance inducibility in S. Choleraesuis. A loss-of-function mutation of marA in a single S. Choleraesuis isolate was created by insertion of the dihydrofolate reductase (DHFR) gene cassette within marA using double homologous recombination. This mutation was complemented with an expression plasmid possessing marA under the control of an IPTG-inducible promoter. Mutation and complementation of marA was verified using polymerase chain reaction, Northern hybridization, and Western blotting assays. Minimum inhibitory concentrations (MICs) of tetracycline, chloramphenicol, nalidixic acid, and rifampin were determined against induced and uninduced wildtype, marA-disrupted and marA-complemented strains using a microbroth dilution assay. Minimum inhibitory concentrations against induced wildtype and marA-complemented strains increased four- to eight-fold for all antimicrobials tested when compared to the uninduced strains while the MICs of the induced marA-disrupted mutant remained the same. However, this increase was abrogated when the cells were grown in the presence of the efflux pump inhibitor compound EPI phe-arg-naphthylamide. The results indicate that a functional marA is solely required for an inducible multiple antimicrobial resistance phenotype in S. Choleraesuis.

Animals↗
PubMed2004

Biodegradability of wheat gluten based bioplastics.

A large variety of wheat gluten based bioplastics, which were plasticized with glycerol, were subjected to biodegradation. The materials covered the total range available for the biochemical control parameter Fi, which expresses the percentage of aggregated proteins. This quantity can be related to the density of covalent crosslinks in the wheat gluten network, which are induced by technological treatments. The biodegradability tests were performed in liquid medium (modified Sturm test) and in farmland soil. All gluten materials were fully degraded after 36 days in aerobic fermentation and within 50 days in farmland soil. No significant differences were observed between the samples. The mineralization half-life time of 3.8 days in the modified Sturm test situated gluten materials among fast degrading polymers. The tests of microbial inhibition experiments revealed no toxic effects of the modified gluten or of its metabolites. Thus, the protein bulk of wheat gluten materials is non-toxic and fully biodegradable, whatever the technological process applied.

Biocompatible Materials↗
PubMed2000

Synthesis and characterization of a novel biodegradable antimicrobial polymer.

Bacterial infection is a frequent complication associated with the use of medical devices. In an effort to address this problem, antibacterial agents have been incorporated or applied directly onto the surfaces of numerous types of medical devices. This study assessed the feasibility of using a novel biodegradable polymer to release antibiotic drugs in response to inflammatory related enzymes. A model drug polymer was synthesized using 1,6-hexane diisocyanate (HDI), polycaprolactone diol (PCL), and a fluoroquinolone antibiotic, ciprofloxacin. Polymers were characterized by size-exclusion chromatography (SEC), and elemental analysis. Biodegradation studies were carried out by incubating the polymers with solutions of cholesterol esterase (CE) or phosphate buffer (pH 7.0) for 30 days at 37 degrees C. The degradation was assessed by high-performance liquid chromatography (HPLC), mass spectrometry (MS) and 14C radiolabel release. Subsequently, the activity of the released antibiotic was assessed against a clinical isolate of Pseudomonas aeruginosa. HPLC analysis showed the release of multiple degradation products which were identified, by tandem MS, to include ciprofloxacin and derivatives of ciprofloxacin. The microbiological assessment showed that the released ciprofloxacin possessed antimicrobial activity; 1 microg/ml was measured after 10 days. The results of this study suggest that these novel bioresponsive antimicrobial polymers or similar analogs show promise for use in the control of medical device associated infections.

Absorbable Implants↗
PubMed1999

Fluoride release, microbial inhibition and microleakage pattern of two orthodontic band cements.

OBJECTIVES: To compare, in vitro, the fluoride release, microbial inhibition and microleakage pattern of a conventional glass ionomer cement (Ketac-Cem) and an acid-modified composite (Ultra Band-Lok) for band cementation. METHODS: Fluoride release was measured from cement discs (3.0 mm diameter and 1.5 mm thick) at regular intervals over 40 days using a potentiometric method. Microbial inhibition was determined for each cement using an agar diffusion test against one of four different strains of Streptococcus mutans. Thirty pairs of banded third molars (15 banded pairs for each cement) were thermocycled and microleakage determined by a dye penetration method. The depth of microleakage was assessed by an index applied by two examiners independently to photographic records taken of the mid-buccal aspect of each tooth. RESULTS: The cumulative and daily fluoride release for days 5, 15 and 40 were significantly greater for Ketac-Cem than for Ultra Band-Lok (all p < 0.05). After the initial set, the anti-microbial activity was significantly greater for Ketac-Cem than for Ultra Band-Lok over the following 24 h period for all four strains of S. mutans (all p < 0.05). There was no significant difference between the two cement groups for microleakage at the cement/enamel interface (p = 0.66) but a borderline significance was detected for microleakage at the cement/band interface (p = 0.051). CONCLUSIONS: The fluoride release and anti-microbial activity of Ketac-Cem were greater than that of Ultra Band-Lok. There was no significant difference in microleakage between the cements at the cement/enamel interface but a borderline difference existed between the cements at the cement/band interface.

Composite Resins↗
PubMed2002

Survey of drug use practices and antibiotic prescribing pattern at a general hospital in Nigeria.

OBJECTIVE: To describe the current drug use practices at the institution, and gather baseline data, which can serve as a basis for designing an appropriate intervention to improve the drug use profile. METHOD: A retrospective audit of in- and out-patient prescriptions, generated during the period January-March 1999. Indicators of drug use pattern include: average number of drugs prescribed per encounter (ANDPE), average number of antibiotics prescribed per encounter (ANAPE), % encounter with antibiotics (PEA), percent of antibiotic prescriptions based on microbial sensitivity test results (MCST). Additionally, a "knowledge, attitude and practice" (KAP) survey of prescribers and dispensers was performed. Indicators of prescribing and dispensing quality include: sources of drug/prescribing information, availability and use of the hospital formulary, knowledge of the prescribing process and the adequacy of the drug supply management system. RESULTS: A total of 9984 outpatient prescriptions and 127 in-patient case notes were audited. The total number of prescribers and dispensers surveyed were 88 and 13 respectively. The ANDPE was found to be 3.16 for out-patients and 9.7 for in-patients, ANAPE was 1.1 and 2.4 for out-patients and inpatients respectively. The PEA was 50.3% for out-patients and 96.7% for in-patients. Only 4.2% of in-patient antibiotic prescriptions were based on MCST and percent encounter with switches in antimicrobial therapy was 52.1% while the average number of switches per encounter was 1.35. In 18.5% of the in-patient encounters there was evidence of drug incompatibilities. The KAP survey revealed that prescribers and dispensers in the hospital rely on different sources for their drug information needs (MIMS vs Martindale Extrapharmacoepia). None of the prescribers surveyed was able to correctly enumerate all the 4 steps involved in the prescribing process, about 25% got at least 2 steps correctly, and only 9.1% of the dispensers surveyed could accurately define a hospital formulary. The drug supply management system was found to be inadequate. CONCLUSION: The survey revealed that appreciable gaps in knowledge with respect to rational drug use, still exists among these cadre of healthcare professionals. The foregoing suggests an urgent need for review of current policies and systems in the hospital with the view of enhancing the drug use practices of the health providers. Specifically it is recommended that there should be an intervention program involving concerted continuing education (to influence the KAP of the various cadre of healthcare providers) and the establishment of a hospital formulary/standard treatment guidelines.

Adolescent↗
PubMed2004

Mode of binding of fibrinogen, fibronectin and iron-binding proteins by animal enterococci.

Sixty-two animal enterococci were examined for their binding of bovine fibrinogen, porcine fibronectin, bovine lactoferrin, bovine apotransferrin and human holotransferrin in the particle agglutination assay (PAA). Individual strains expressed binding of selected glycoproteins to various degrees (0, 1, 2, 3), whereas bovine fibrinogen binding of enterococci from goats, rabbits and rodents was the strongest (3) in general. Porcine fibronectin was bound weakly (1 or 2) by enterococci from horses, dogs, poultry, rabbits and rodents, while most of the goat isolates and half of the dog feed isolates did not bind fibronectin (0). Bovine lactoferrin was bound especially by the isolates from rodents and rabbits. Bovine apotransferrin was bound very weakly (1) by only a few isolates. Human holotransferrin was bound to a greater extent than apotransferrin by some isolates from rabbits and rodents. Since multiresistant strains are preferred in our binding studies, enterococci were also examined for their antibiotic resistance pattern. Almost all investigated isolates were resistant at least to one antibiotic. However, some strains displayed resistance to five or six antibiotics of 10 antibiotics tested. In a study of the inhibitory effect of heparin, porcine mucin and hyaluronic acid, the greatest effect was observed after heparin treatment of bacterial cells. These observations, as well as the expression of heparin binding by most strains, may suggest that at least one mode of enterococcal attachment utilizes glycosaminoglycan chains present on the surface of adherent cells.

Animals↗
PubMed2006

Small molecules with antimicrobial activity against E. coli and P. aeruginosa identified by high-throughput screening.

BACKGROUND AND PURPOSE: New antimicrobials are needed because of the emergence of organisms that are resistant to available antimicrobials. The purpose of this study was to evaluate a high-throughput screening approach to identify antibacterials against two common disease-causing bacteria, and to determine the frequency, novelty, and potency of compounds with antibacterial activity. EXPERIMENTAL APPROACH: A high-throughput, turbidometric assay of bacterial growth in a 96-well plate format was used to screen a diverse collection of 150,000 small molecules for antibacterial activity against E. coli and P. aeruginosa. The statistical Z'-factor for the assay was > or = 0.7. KEY RESULTS: Screening for inhibition of E. coli growth gave a 'hit' rate (> 60% inhibition at 12.5 microM) of 0.025%, which was more than 5-fold reduced for P. aeruginosa. The most potent antibacterials (EC50 < 0.5 microM) were of the nitrofuran class followed by naphthalimide, salicylanilide, bipyridinium and quinoazolinediamine chemical classes. Screening of > 250 analogs of the most potent antibacterial classes established structure-activity data sets. CONCLUSIONS AND IMPLICATIONS: Our results validate and demonstrate the utility of a growth-based phenotype screen for rapid identification of small-molecule antibacterials. The favourable efficacy and structure-activity data for several of the antibacterial classes suggests their potential development for clinical use.

Anti-Bacterial Agents↗
PubMed2000

Temperature-regulated efflux pump/potassium antiporter system mediates resistance to cationic antimicrobial peptides in Yersinia.

Most bacterial pathogens are resistant to cationic antimicrobial peptides (CAMPs) that are key components of the innate immunity of both vertebrates and invertebrates. In Gram-negative bacteria, the known CAMPs resistance mechanisms involve outer membrane (OM) modifications and specifically those in the lipopolysaccharide (LPS) molecule. Here we report, the characterization of a novel CAMPs resistance mechanism present in Yersinia that is dependent on an efflux pump/potassium antiporter system formed by the RosA and RosB proteins. The RosA/RosB system is activated by a temperature shift to 37 degrees C, but is also induced by the presence of the CAMPs, such as polymyxin B. This is the first report of a CAMPs resistance system that is induced by the presence of CAMPs. It is proposed that the RosA/RosB system protects the bacteria by both acidifying the cytoplasm to prevent the CAMPs action and pumping the CAMPs out of the cell.

Anti-Bacterial Agents↗
PubMed2001

Expression of the Pho regulon negatively regulates biofilm formation by Pseudomonas aureofaciens PA147-2.

We report the isolation of insertional mutations to the pstC and pstA genes of the phosphate-specific transport (pst) operon that results in loss of biofilm formation by Pseudomonas aureofaciens PA147-2. Consistent with the known roles of the Pst system in Escherichia coli and Pseudomonas aeruginosa, both P. aureofaciens pst mutants were demonstrated to have defects in inorganic phosphate (P(i)) transport and repression of Pho regulon expression. Subsequently, biofilm formation by the wild type was shown to require a threshold concentration of extracellular P(i). The two-component regulatory pair PhoR/PhoB is responsible for upregulation of Pho regulon expression in response to P(i)-limiting environments. By generating phoR mutants that were unable to express the Pho regulon, we were able to restore biofilm formation by P. aureofaciens in P(i)-limiting conditions. This result suggests that gene(s) within the Pho regulon act to regulate biofilm formation negatively in low-P(i) environments, and that phoR mutations uncouple PA147-2 from such regulatory constraints. Furthermore, the inability of pst mutants to repress Pho regulon expression accounts for their inability to form biofilms in non-limiting P(i) environments. Preliminary evidence suggests that the Pst system is also required for antifungal activity by PA147-2. During phenotypic analysis of pst mutants, we also uncovered novelties in relation to P(i) assimilation and Pho regulon control in P. aureofaciens.

ATP-Binding Cassette Transporters↗
PubMed2002

Helichrysum italicum extract interferes with the production of enterotoxins by Staphylococcus aureus.

AIMS: The purpose of this work was to evaluate the effect of Helichrysum italicum extract on enterotoxin (A-D) production by Staphylococcus aureus strains. METHODS AND RESULTS: The production of enterotoxins A-D in the presence or absence of H.italicum diethyl ether extract was estimated in microtiter plates using a reversed passive latex agglutination (SET-RPLA) kit (Oxoid, Basingstoke, UK). The results indicate that, in culture medium, inhibition of staphylococcal growth and enterotoxins appeared with 250-125 microg ml(-1) of the extract. Lower concentrations of the extract (62.5-31.25 microg ml(-1)) did not affect the final viable count of Staph. aureus but reduced the production of enterotoxins B and C. CONCLUSIONS: H. italicum interferes with growth and production of enterotoxins by Staph. aureus. SIGNIFICANCE AND IMPACT OF THE STUDY: There is considerable interest in the use of natural compounds as alternative methods to control undesirable pathogenic micro-organisms.

Asteraceae↗