Contact Killing Research Articles

Adhesion and Viability of Two Enterococcal Strains on Covalently Grafted Chitosan and Chitosan/κ-Carrageenan Multilayers

Chitosans are natural aminopolysaccharides, whose low cytotoxicity suggests their potential use for nonadhesive, antibacterial coatings on biomaterials implant surfaces. Here, the antiadhesive behavior and ability to kill bacteria upon adhesion ("contact killing") of chitosan coatings were evaluated for two strains of Enterococcus faecalis, isolated from clogged biliary stents. Chitosan coatings covalently grafted or applied as chitosan/kappa-carrageenan multilayers were characterized by ellipsometry, scanning force microscopy (SFM), X-ray photoelectron spectroscopy (XPS), and electrokinetic measurements. Decreases in initial bacterial deposition rates and the number of bacteria adhering in a more advanced state of the adhesion process were observed on both types of modified surfaces, with more pronounced effects on highly hydrated multilayers. Adhesion of negatively charged enterococci was slightly enhanced on chitosan-terminated multilayers, but antibacterial effect was absent on kappa-carrageenan-terminated multilayers. Thus, the efficacy of multilayers remains an interesting interplay between the promoting effect of cationically charged groups on adhesion of negatively charged bacteria and, on the other hand, their antibacterial effects.

Adhesion and viability of waterborne pathogens on p‐DADMAC coatings

The attachment of waterborne pathogens onto surfaces can be increased by coating the surfaces with positive charge-enhancing polymers. In this paper, the increased efficacy of polydiallyldimethylammonium chloride (p-DADMAC) coatings on glass was evaluated in a parallel plate flow chamber with the use of waterborne pathogens (Raoultella terrigena, Escherichia coli, and Brevundimonas diminuta). p-DADMAC coatings strongly compensated the highly negative charges on the glass surface and even yielded a positively charged surface when applied from a 500 ppm solution. Whereas none of the strains adhered from water to glass due to electrostatic repulsion, R. terrigena and E. coli readily adhered in high numbers to p-DADMAC coated glass slides applied from 1, 100, or 500 ppm aqueous solutions. B. diminuta only adhered to a positively charged p-DADMAC coating applied from a 500 ppm solution. In addition, all p-DADMAC coatings indicated strong contact killing with the bacterial species used in this study by live/dead staining techniques. In summary, this paper demonstrates the potential of p-DADMAC coatings to strongly enhance bacterial adhesion. Moreover, once adhered, bacterial viability can be reduced by the positively charged ammonium groups in the coating.

Contact killing antimicrobial acrylic bone cements: preparation and characterization

Novel antimicrobial poly(methyl methacrylate) (PMMA)-based bone cement was synthesized by co-polymerizing PMMA/MMA with various percentages of quaternary amine dimethacrylate (QADMA) by free radical bulk polymerization technique at room temperature using benzoyl peroxide and N, N-dimethyl-p-toulidine (DMPT) as a redox initiator. The modified bone cement was characterized by FT-IR and 1H-NMR spectral studies. The thermal and physical properties of the bone cements of varying composition of QADMA were evaluated by thermogravimetric analysis (TGA), differential calorimetry (DSC) and contact angle measurements. Peak exothermic temperature was observed to decrease, while setting time increased with increase in QADMA content in the bone cement formulations. The antibacterial activity of the synthesized bone cement containing quaternary amine dimethacrylate against Escherichia coli and Staphylococcus aureus was studied by zone of inhibition, colony count method and scanning electron microscopy (SEM). QADMA containing acrylic bone cement showed a broad spectrum of contact killing antimicrobial properties. Retention of E. coli onto the surface of PMMA bone cement was observed, whereas there was complete prevention of retention of E. coli onto the modified PMMA bone cement with 15% QADMA. The studies were compared with the acrylic bone cement synthesized using 15% N-vinyl-2-pyrrolidone (NVP) in place of QADMA to which iodine was added as an antimicrobial agent during co-polymerization.

Preparation of iodine containing quaternary amine methacrylate copolymers and their contact killing antimicrobial properties

AbstractA novel quaternary amine methacrylate monomer (QAMA) was synthesized by amination of dimethacrylate with piperazine followed by its quaternization using an alkyl iodide. Copolymerization of QAMA with 2‐hydroxyethyl methacrylate was carried out by free radical bulk polymerization technique at room temperature using ammonium persulfate and N,N,N′,N′‐tetramethyl ethylenediamine as a redox initiator. The monomer as well as copolymers was characterized by FTIR and 1H NMR spectral studies. Thermal and physical characteristics of copolymers of varying compositions of QAMA were evaluated by thermogravimetric analysis, differential calorimetry, contact angle and scanning electron microscopy. The antibacterial activity of the synthesized quaternary amine dimethacrylate copolymers against Escherichia coli and Staphylococcus aureus was studied by zone of inhibition and colony count method. QAMA copolymers showed broad‐spectrum contact killing antibacterial properties without releasing any active agent as checked by iodide‐selective ion meter. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 102: 1038–1044, 2006

Differential Inhibition of RAW264.7 Macrophage Tumoricidal Activity by 9Tetrahydrocannabinol

delta 9tetrahydrocannabinol (THC), the major psychoactive component of marijuana, has been shown to inhibit macrophage cell contact-dependent cytolysis of tumor cells. The purpose of this study was to determine whether THC inhibited macrophage cytolytic function by targeting selectively tumor necrosis factor (TNF)-dependent pathways versus L-arginine-dependent reactive nitrogen intermediates. An in vitro system employing RAW264.7 macrophage-like cells as effectors and TNF-sensitive mouse L929 fibroblasts or nitric oxide (NO.)-sensitive P815 mastocytoma cells as targets, was employed to assess the effect of THC on cytolysis. Macrophages were pretreated with THC or vehicle for 48 hr, subjected to multistep activation with 10 U/ml recombinant mouse gamma-interferon (IFN-gamma) plus 100 ng/ml LPS or to direct activation with 1 microgram/ml LPS, and co-cultured with tumor cells in the presence or absence of THC. THC inhibited TNF-dependent killing by macrophages subjected to either multistep or direct activation. Decreased amounts of TNF-alpha were detected in medium of macrophage cultures treated with THC. In contrast, THC inhibited NO.-dependent cell contact killing only for macrophages subjected to direct activation. Decreased levels of NO2-, a stable degradation product of the short-lived and highly toxic effector molecule NO., were produced by these macrophages. In addition, the effect of the enantiomeric pairs (-)CP55,940/(+)CP56,667 or (-)HU-210/(+)HU-211 on macrophage cell contact-dependent killing was assessed. Inhibition of macrophage tumoricidal activity against TNF-sensitive L929 cells was effected by both isomers of THC analogs. In contrast, both of the enantiomeric pairs had an effect on killing of NO.-sensitive P815 mastocytoma cells only for macrophages subjected to direct activation. These data suggest that cannabinoids inhibit macrophage cell contact-dependent killing of tumor cells by a noncannabinoid receptor-mediated mechanism. However, specific cytolytic pathways are inhibited differentially by cannabinoids depending on the activation stimuli to which macrophages are exposed.

Insecticidal activity of pyrrolizine derivatives isolated fromStreptomyces griseus

Mixtures of 5,6,7,7a-tetrahydro-3H-pyrrolizin-3-one (1) and 5,6,7,7a-tetrahydro-3H-pyrrolizin-7a-ol-3-one (2) isolated from shaken cultures ofStreptomyces griseus exhibited contact killing effects on larvae ofLeptinotarsa decemlineata andAedes aegypti comparable with those of macrotetrolides and the commercial bioinsecticide Bactoculicid, respectively.Galleria mellonella (caterpillars),Anagasta kuhniella andTetranychus urticae were insensitive to this action.

Ultrastructural features of the lymphocyte-stimulated halos produced by human glioma-derived cells in vitro

Many glioma-derived cell lines have the capability of escaping cell-mediated immune attack. One mechanism of escape is the secretion of a hyaluronidase-sensitive mucopolysaccharide coat by these cells. This coat prevents contact and tumor cell killing by specific cytolytic allogeneic lymphocytes. The production of the coat by the tumor cells is stimulated by a macromolecular factor released by peripheral blood mononuclear (PBMC) cells in culture. We have examined the morphologic and ultrastructural features of this extracellular matrix. Three coat-producing lines were studied. Under phase contrast light microscopy, the coat is a clear pericellular 'halo'. To stain this zone, ruthenium red and Alcian Blue 8 G stains, which bind to acid mucopolysaccharides (to a large extent, hyaluronic acid), were used. The two stains produced similar results. With light microscopy, a weblike pattern of stain was evident throughout the halo region. With transmission electron microscopy, staining was found along the plasma membrane of the glioma cells and their microvilli, stretching in long, branching filaments from these surfaces and, in some instances, from one microvillus to the next. Since mucopolysaccharide matrices have a large aqueous component, it was necessary to determine whether dehydration alters the stain pattern. Therefore, undehydrated ruthenium red stained specimens from each culture were embedded in Quetal 651 (Ted Pella, Inc., Tustin, CA), a water soluble plastic. No morphologic differences were noted between the hydrated and dehydrated specimens. This study indicates that numerous long microvilli and a secreted mucopolysaccharide matrix are important structural elements of the lymphocyte-stimulated tumor cell halo in vitro. The mechanism by which the PBMC factor stimulates coat formation and the importance of the coat in in vivo tumor defenses remain to be elucidated.

T cell subsets in allograft rejection. In situ characterization of T cell subsets in human skin allografts by the use of monoclonal antibodies.

We have provided evidence that both major T cell subsets, T4-positive (helper/inducer) and T8-positive (cytotoxic/suppressor), infiltrate human skin allografts. Overall, and in the graft dermis and graft bed, T4-positive cells were predominant (1.5 to 3 times more numerous than T8-positive cells). In contrast, T8-positive cells were relatively more numerous in the epidermis and hair follicles. Rejection probably proceeded by two apparently independent pathways: 1) direct contact killing of graft epithelial cells, presumably by immunologically specific T8-positive cytotoxic cells, and 2) injury of microvascular endothelium of both the graft and graft bed with secondary graft infarction. Although important in first set skin allograft rejection, the mechanism of the second type of killing is uncertain. T4-positive cells were probably involved, as evidenced by their greater numbers; furthermore, studies in mice have shown that transfused helper/inducer cells are able to effect first-set skin graft rejection. It remains to be determined whether T4-positive cells act alone or cooperate with other cells to destroy vessels and bring about graft rejection. Langerhans cells were recognized in epithelial and dermal compartments of both allografts and autografts by their reactivity with anti-T6 and anti-Ia antibodies. We could not determine whether such cells in allografts were of host or donor origin.

The influence of suspending agents on preservative activity in aqueous solid/liquid dispersions

An investigation has been made of the effect of particulate solids on the activity of selected antimicrobial preservatives in the absence and presence of pharmaceutical suspending agents. Certain preservatives were adversely affected by the suspended systems, as shown by microbiological assay, using contact killing times.

The activity of some pyrethroids against Periplaneta americana and Blattella germanica

AbstractThe knockdown and contact killing actions of various pyrethroids were compared using Blattella germanica and Periplaneta americana. A wide range of knockdown activity was found; 5‐benzyl‐3‐furylmethyl (1R)‐cis‐3‐(dihydro‐2‐oxo‐3‐thienylidenemethyl)‐2,2‐dimethylcyclopropanecarboxylate (RU 15525) acted fastest, more rapidly than pyrethrins, against B. germanica as well as having a low LD50 value. Topical application and direct spray tests showed that (S)‐α‐cyano‐3‐phenoxybenzyl (1R)‐cir‐3‐(2,2‐dibromovinyl)‐2,2‐dimethylcyclopropanecarboxylate (NRDC 161) was more active as a killing agent, by an order of magnitude, than cismethrin, the next most active compound, and also had considerable knockdown activity. Piperonyl butoxide generally had little synergist effect. Female P. americana were over three times more tolerant than males to a range of insecticides applied topically. Residual knockdown action in the WHO resistance test was observed to provide baseline data. There was little overlap in speed of action between pyrethroids and other insecticides among the compounds tested.

Herbicidins A and B, two new antibiotics with herbicidal activity. I. Producing organism and biological activities.

Herbicidins A and B, two new antibiotics with selective and contact herbicidal activity, were produced by a new species of Streptomyces designated as S. sagononensis. Among tested microorganisms, herbicidins indicated some activity against fungi in vitro and Xanthomonas oryzae in vivo. Their characteristic features were brought into relief by their selective and contact killing effect on many dicotyledonous plant as well as their inhibition of germination of the plant seeds, such as Chinese cabbage.

Transplantation and immunoincompatibility reactions among reef-building corals

Intracolony or syngeneic transplants of pieces of living coral, either occurring naturally on reefs or produced experimentally, were consistently compatible. In contrast, intercolony allografts performed extensively inAcropora formosa and inPorites andrewsii were incompatible, as were allografts occurring naturally in additional species observed in reef communities. Diverse interspecific combinations of coral xenografts generally displayed greater incompatibilities. Four levels of immuno-reactivity, with manifestations ranging from mild to severe, were distinguishable as follows: a) contact avoidance reactions, b) allogeneic contact incompatibility, c) chronic xenogeneic incompatibility, and d) acute interspecific aggression.Allogeneic incompatibility, most studied inA. formosa, was characterized by soft tissue contact avoidance wherever possible; absence of tissue death despite extensive enforced contact (18–20 days); and late interfacial cementation terminating allogeneic soft tissue contact. The properties ofchronic xenogeneic incompatibility as found in diverse species combinations, are slow onset (>7 days), bidirectional or unidirectional killing in contact zones, and short-range or localized effectiveness.Acute interspecific aggression initiated byFungia fungites as a dominant “aggressor ” is distinguished by early occurrence (2–7 days) and unidirectional contact killing. The presence or absence of these acute xenogeneic reactions sharply discriminated between species within each of the generaAcropora, Pocillopora, andPorites. No hierarchy of aggressive interactions was evident at the generic level.

Membrane Filter Technique for Testing the Bactericidal Activity of Iodophors

A method has been devised for determining the contact killing time of a water-soluble antiseptic or disinfectant agent using a membrane filter as the test organism carrier. The method is useful in comparing the bactericidal activity of different concentrations of an agent since the number of organisms killed following a specific contact time can readily be determined on a percentage basis compared to that of a control. The problems of bacterial clumping, “wild plusses,” and carry-over bacteriostasis are eliminated. A suspension of the test organism was filtered through a membrane filter, a solution of the agent was added, was allowed to remain in contact with the test organism for a specific time, and was filtered. The treated membrane was washed with a neutralize, was rinsed, and was incubated on nutrient agar. The number of surviving organisms, if any, was determined by colony count. Solutions of povidone-iodine as well as commercial products containing it were tested against Staphylococcus aureus, FDA 209, ATCC 6538. Solutions containing a 1:15,000 concentration of available iodine gave a kill of 99.91 percent following a 1-min. contact.