In Vitro Evaluation of the Efficacy of a Silver-Coated Catheter

Bacteria commonly associated with nosocomial urinary tract infections were examined in vitro for their relative adherence to latex, 100% silicone-, hydrogel-coated latex-, and hydrogel/silver-coated latex urinary catheters. Degrees of adherence within 2 h were determined with cells radiolabeled with leucine. Adherence was greatest and equivalent on silicone and latex catheters. Adherence of four strains of Escherichia coli to the hydrogel/silver-coated catheter was decreased by 50% to 99% in comparison with the other catheters. Repeat testing with strains of E. coli and Pseudomonas aeruginosa with over 50 catheters demonstrated a consistency in the inhibition. The hydrophilic coating of the catheter appeared to be primary in the decreased adherence phenomenon followed by a secondary biocidal effect of the silver ion. Received: 2 December 1995 / Accepted: 3 January 1996     

Effects of silver on adherence of bacteria to urinary catheters: In vitro studies

Strains of Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Enterococcus faecalis, and Klebsiella pneumoniae, mostly from complicated urinary tract infections, showed reduced adherence to silver-treated silicone or latex catheters as compared with latex or silicone catheters. The relative degrees of cell adherence to catheters at 2 h or 18 h, as indicated by radiolabeled cell assays, were in general agreement with growth rate-reduction assays and scanning-electron-microscopy data. For strains of E. coli, the correlation between cell hydrophobicity and degree of adherence to catheters was not significant. Antibiotic resistance (tetracycline, sulfathiazine, neomycin, kanamycin) and silver resistance were not associated. The radiolabel adherence procedure provided a quantitative method for evaluating the relative antimicrobial efficacy of silver-treated catheters.     

Evaluation of the Adherence of Candida Species to Urinary Catheters

Catheter-associated urinary tract infections (CAUTIs) are the most common kind of nosocomial infection. Recent years have seen a significant increase in numbers of infections caused by yeasts of the genus Candida. The adherence of a microorganism to the host surface is a decisive factor in the success of colonization and the pathogenesis of infection. The objective of this work was to evaluate the adherence of species of the genus Candida to urinary catheters. In vitro adherence to the sections of latex and silicon catheters of Candida albicans and Candida parapsilosis were studied. Adherence was measured by counting the number of adhering viable cells and the results were expressed as Colonies Forming Units per mL. The results demonstrated that the latex catheter facilitated adherence more than the silicon catheter (p < 0.01). The adherence of the C. albicans was significantly greater than C. parapsilosis on latex, but it was similar on silicon.     

De novo induction of island capsule flap by using two silastic sheets: Part 1. Generation

A new experimental model for de novo generation of an axial pattern island flap has been designed in a rat model. The purpose of this study was to make a sufficient vascular carrier, as an island capsule flap, with only vascular pedicles and addition of collagen fibers induced by foreign-body reaction. The femoral arteriovenous bundle was isolated and sandwiched between two 2.5 x 1.5 cm Silastic sheets. Eight weeks later, as a delay procedure, femoral vessels were ligated at the distal end of the Silastic sheets and the four margins of the sheets were divided except for the vascular pedicle. This capsule flap was raised as a secondary island flap connected only by its vascular pedicle, then it was sutured back in place. Ten days after the delay procedure, the upper Silastic sheet was removed and a full-thickness skin graft was performed on the capsular island flap. Animals were killed at 80 days. A total of 40 axial pattern capsulocutaneous flaps from 20 Sprague-Dawley rats were successfully achieved. Pathologic study revealed neovascularization, and abundantly impregnated vascular structures near the pedicle were observed with randomly developed collagen fibers. The skin graft took 100 percent on this newly formed capsular flap; therefore, the capsule structure was able to survive on its own and support skin grafts. This experiment, by using an isolated femoral artery and vein as the main pedicle, led to the formation of a capsule flap through a normal foreign body reaction between two Silastic sheet implants. This new flap can be used as a reliable vascular carrier for various needs with minimal donor morbidity.     

Citropin 1.1-treated central venous catheters improve the efficacy of hydrophobic antibiotics in the treatment of experimental staphylococcal catheter-related infection

An in vitro antibiotic susceptibility assay for Staphylococcus aureus biofilms developed on 96-well polystyrene tissue culture plates was performed to elucidate the activity of citropin 1.1, rifampin and minocycline. Efficacy studies were performed in a rat model of staphylococcal CVC infection. Silastic catheters were implanted into the superior cava. Twenty-four hours after implantation the catheters were filled with citropin 1.1 (10 microg/mL). Thirty minutes later the rats were challenged via the CVC with 1.0 x 10(6) CFU of S. aureus strain Smith diffuse. Administration of antibiotics into the CVC (the antibiotic lock technique) began 24 h later. The study included: one control group (no CVC infection), one contaminated group that did not receive any antibiotic prophylaxis, one contaminated group that received citropin 1.1-treated CVC, two contaminated groups that received citropin 1.1-treated CVC plus rifampin and minocycline at concentrations equal to MBCs for adherent cells and 1024 microg/mL in a volume of 0.1 mL that filled the CVC and two contaminated groups that received rifampin or minocycline at the same concentrations. All catheters were explanted 7 days after implantation. Main outcome measures were: minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), synergy studies, quantitative culture of the biofilm formed on the catheters and surrounding venous tissues, and quantitative peripheral blood cultures. MICs of conventional antibiotics against the bacteria in a biofilm were at least four-fold higher than against the freely growing planktonic cells. In contrast, when antibiotics were used on citropin 1.1 pre-treated cells they showed comparable activity against both biofilm and planktonic organisms. The in vivo studies show that when CVCs were pre-treated with citropin 1.1 or with a high dose of antibiotics, biofilm bacterial load was reduced from 10(7) to 10(3) CFU/mL and bacteremia reduced from 10(3) to 10(1) CFU/mL. When CVCs were treated both with citropin 1.1 and antibiotics, biofilm bacterial load was further reduced to 10(1) CFU/mL and bacteremia was not detected, suggesting 100% elimination of bacteremia and a log 6 reduction in biofilm load. Citropin 1.1 significantly reduces bacterial load and enhances the effect of hydrophobic antibiotics in the treatment of CVC-associated S. aureus infections.     

A trial application of the latex test for evaluation of antibody level in rabbit immune sera]

A latex test was elaborated which served for evaluation of quality of rabbit immune sera for antigen 0 of selected Gram-negative bacteria. Sensitivity and specificity of this test in comparison with passive hemagglutination and immunoenzymatic DOT-ELISA reactions was evaluated. These studies were performed on immune sera for antigen O of Salmonella groups B, C1, C2, D and E, Yersinia pseudotuberculosis and in antigen preparations for above listed microorganisms both in homologous and heterologous systems. It was found that sensitivity of the latex test is 9 to 160 times lower than that of passive hemagglutination and 7 to 307 lower than for DOT-ELISA. Sensitivity of the latex test and passive hemagglutination reaction was evaluated on the basis of results of cross reaction between studied antigens and unabsorbed rabbit sera, establishing so called sensitivity indexes, which were informing how many times heterologous titer is lower than homologous titer. So evaluated sensitivity of the latex test was close to sensitivity of the passive hemagglutination reaction. It was found that slide latex test is characterized by satisfactory sensitivity and good sensitiveness and may be used for evaluation of antibody level 0 antigens of Salmonella and Yersinia. The value of this test is characterized by high repeatability of results, as well as low work and time-consuming.     

The problem of catheter encrustation

Catheter encrustation was studied in a group of long-stay hospital patients using both latex and silicone catheters. Moisture accounted for 80% by weight of the encrusted material with both catheters. Of the dry weight 90% was composed of protein, calcium, phosphorus, magnesium and uric acid. No relationship was found between the amounts of these substances in the urine and in the encrusted material. High levels of calcium, phosphorus and magnesium were found in the encrusted material from patients infected with Proteus organisms. No direct relationship was found between the duration of catheter drainage and the degree of encrustation, and there was a variation in patient susceptibility to encrustation irrespective of the catheter material. There was significantly less encrustation associated with silicone catheters.     

Recognition of target colloid species by conjugates of a linear polyelectrolyte with a vector protein

Redistribution reaction of quaternized poly-4-vinylpyridine polycations and their conjugates with alpha-chymotrypsin by oppositely charged latex particles is disclosed. The polycations are strongly adsorbed on the latex surface. Nevertheless, they are able to migrate between the latex species via occasional interparticle contacts. In the case of a homogeneous latex such interchange results in uniform distribution of polycations by latex particles. The distribution drastically changes, when alpha-chymotrypsin-polycation conjugates interact with a mixture of two latexes: one chemically modified by bovine serum albumin and the other one by specific protein inhibitor of alpha-chymotrypsin. In this case the interchanging polycations are finally fixed on the latex particles carrying the centres of specific binding of the enzyme vector, i.e. recognize them in the latex mixture. The obtained results are considered to mimic physico-chemical interaction and recognition of target supermolecular bio-objects by large macromolecules carrying relatively small molecular vectors.     

Uropathogenic Escherichia coli adhere to urinary catheters without using fimbriae

Abstract Five well-characterized urinary and fecal isolates of Escherichia coli were found to be hydrophilic irrespective of their serotypes and their ability to express fimbriae. All the strains were able to adhere to silicone latex urinary catheters, although strain 917, which expressed type P fimbriae as its only adhesin, adhered poorly. Although specific adhesins, particularly fimbriae, have been shown to mediate adhesion of E. coli to uroepithelial cells, they do not mediate specific adhesion onto urinary catheter material. The overall surfaces of the strains, tested using microelectrophoresis as a function of pH and X-ray photoelectron spectroscopy, were not significantly different, thus suggesting more non-specific adhesion mechanisms to urinary catheters.     

Effects of Hydrogel/Silver Coatings on In Vitro Adhesion to Catheters of Bacteria Associated with Urinary Tract Infections

Sections of sterile all-silicone-, hydrogel/silver-all-silicone-, and hydrogel/silver-latex-Foley urinary catheters were exposed to suspensions of bacteria and Candida albicans associated with urinary tract infections. The adhesion of these microorganisms to the catheters was determined with a radiolabel–cell procedure and scanning electron microscopy. Anomalous data with the radiolabel procedure were produced with the hydrogel/silver-latex catheters for certain species. These aberrant data were related to adhesion on the untreated cut ends of the latex catheter. Radiolabel-cell-adhesion procedures that involve sections of coated materials may need to be supplemented with additional procedures such as scanning electron microscopy for valid interpretations of the data. Adhesion to the hydrogel/silver catheters by both Gram-positive- and Gram-negative bacteria most commonly associated with nosocomial urinary tract infections, including a strain of Pseudomonas aeruginosa noted for its superior adhesion capacity, was significantly lower than the adhesion to the control all-silicone catheter. Received: 21 January 2000 / Accepted: 26 February 2000     

Polystyrene microspheres as bioligand carriers in the determination of Δ-9-tetrahydrocannabinol in human urine

For the first time, a diagnostic test system for the determination of drugs, in particular, cannabinoids, in human urine was constructed on the basis of the latex agglutination reaction. For this purpose, polystyrene microspheres with a diameter of 0.6 μm and a narrow particle size distribution and carboxylic groups of stabilizer molecules on their surfaces were used. The studies revealed the optimal characteristics of particle suspension, which provide for a high sensitivity and specificity level of the test, in particular, the quantitative range of the antibody content for covalent binding with carboxylic groups on the surface of polystyrene microspheres. The operating titer of Δ-9-tetrahydrocannabinol (Δ-9-THC)-specific antibodies was 1/16-1/512 for a conjugated antigen taken at a concentration of 0.1 mg/mL. The sensitivity of the latex agglutination reaction when determining Δ-9-THC was 150 ng/mL.     

Identification and characterization of latex-specific proteins in opium poppy

Latex from the opium poppy, Papaver somniferum L., was analyzed by polyacrylamide gel electrophoresis (PAGE). Two latex-specific bands were identified in protein samples of poppy latex using one-dimensional native PAGE. Second dimension analysis with SDS-PAGE indicates that these proteins have a relative molecular weight of approximately 20 kilodaltons. We have termed these polypeptides the major latex proteins (MLPs). Polyclonal antibodies prepared against the MLPs were used to probe protein gel blots of latex and poppy tissues known to lack laticifers. Laticifer-free tissues showed no reaction with anti-MLP immunoglobulin G indicating that MLPs are found only in poppy latex. MLP distribution was also examined in mature opium poppy tissues by immunocytochemistry. Laticifers were differentially labeled by fluorescein isothiocyanate secondary labeling of anti-MLP immunoglobulin G and could easily be identified in both transverse and longitudinal section. Fractionation studies of isolated latex showed that MLPs are concentrated in the latex cytosol and not in alkaloidal vesicles. Analysis of latex proteins by conventional two-dimensional electrophoresis indicates that the two MLP bands are composed of several distinct polypeptides with similar relative molecular weights. The pIs of these molecules range from 6.0 to 3.5. The role(s) of MLPs in laticifer metabolism has not been determined.     

Regulatory initiatives for natural latex allergy: US perspectives

The US Food and Drug Administration (FDA) has regulatory authority over foods, human drugs, cosmetics, medical devices, radiological products, biologics, and veterinary products. Among these products, FDA believes that the use of medical devices, including medical gloves, condoms, catheters, and breathing bags, represents the greatest source of natural latex proteins to exposed individuals. A medical device is defined in the Federal Food Drug and Cosmetic Act (FFDCA) as an instrument, apparatus, implement, machine, etc., that is intended for use in the diagnosis or treatment of disease or is intended to affect the structure or any function of the body of a human or other animal, and that does not achieve any of its principal intended purposes through chemical action in the body. This article provides some brief, general background about FDA's medical device regulatory process and then addresses the issue of natural latex allergy. Finally we discuss the steps the Agency has taken to evaluate the magnitude and nature of the problem, and FDA's efforts to assist manufacturers, health professionals, and others in minimizing exposure and sensitization to natural latex proteins in medical devices.     

The effect of chlorhexidine and gentian violet on the adherence of <Emphasis Type="Italic">Candida</Emphasis> spp. to urinary catheters

Urinary tract infection associated with catheters is the most common infection in the hospital environment. The adherence of microorganisms to the surface is a determining factor in colonization and infection. Antiseptics such as chlorhexidine and gentian violet have been shown to be effective against yeasts, as well as having low toxicity and being low-cost. The objective of the present study was to evaluate whether prior treatment of siliconized latex urinary catheters with antiseptics reduces the adherence of yeasts. Two reference strains of C. albicans (ATCC 645448 and ATCC 90028) and six strains isolated from catheter, two each of C. albicans, C. tropicalis, and C. parapsilosis, were used. An in␣vitro study of adherence was carried out with previously treated catheters, in separate experiments of 1 h and 24 h of incubation under continued shaking. The relative hydrophobicity of the cell surface of the yeasts before and after 1 h of exposure to chlorhexidine was determined. The results demonstrated that both treatments were effective in controlling the adherence of yeast to the catheter (P < 0.0001), and that the hydrophobicity of the eight strains significantly increased after contact with chlorhexidine (P < 0.0001). These results suggest that the antimicrobial activity of chlorhexidine and gentian violet reduces the adherence of the microorganisms to the catheter.     

Síndrome de la bolsa de orina púrpura en dos pacientes institucionalizados

Purple urine bag syndrome (PUBS) is an uncommon but particularly striking phenomenon characterised by a chemical reaction involving the urine, plastic and certain enzymes from some sulphatase- and phosphatase-producing bacteria, including Proteus mirabilis, Escherichia coli and Morganella morganii, amongst others. Following this reaction, the catheter and the bag may be stained red, blue or purple. This phenomenon tends to occur in patients with multiple pathology and with urinary catheters, as part of a urinary tract infection. We describe two clinical cases of PUBS in institutionalised patients with permanent urinary catheters.     

OBITUARIES

A study of 300 patients receiving intravenous therapy showed that 90 had associated phlebitis. Because of this high rate of complications, the use of long plastic catheters, with the tip located in a large central vessel, was investigated. One hundred and one catheters were inserted into the basilic vein through a cut-down. The patients were divided into four groups: infusions lasting one to seven days, eight to 14 days, 15 to 28 days and 29 days or longer. The most common complication was obstruction of the catheter with clotted blood. In four patients the catheters had to be removed because of phlebitis; two were pulled out by the patients themselves. Infection was not observed. Two factors probably contributed to the successful infusions: the composition of the plastic catheters (nylon) and the location of the tip in a large central vessel.     

Conventional detection method of fibrinogen and fibrin degradation products using latex piezoelectric immunoassay

We developed a conventional immunosensor for fibrinogen and fibrin degradation products (FDP) to combine a quartz crystal microbalance (QCM) with the agglutination reaction of immunized latex beads. FDP induced an immunoreaction due to anti-FDP antibody immobilized latex particles. We successfully measured FDP concentration of in human serum within 10 min by QCM method. The detection range of QCM immunosensor is covered with screening concentration of FDP in serum (<10 microg/ml of FDP). The time course of latex agglutination obtained from QCM immunosensor is synchronized to that of latex photometric immunoassay. SEM was used to observe the surface of QCM that applied FDP serum. The size of latex particles agglutinated on the QCM electrode increased concomitant with FDP concentration. Frequency shift on immunoreaction explains the increased adsorption amount of agglutinated latex on QCM.     

Heterophile infectious mononucleosis antigen used in the latex diagnostic test. I. A method of antigen isolation and its serologic activity

The aim of this study was to elaborate a method of heterophile mononucleosis antigen preparation useful for latex coating. This antigen was isolated from bovine red blood cells stroma by the technique of Schwarzweiss and Tomcsik with author's own modification, in which introductory extraction of erythrocytes stroma ++ was performed by means of trichloracetic acid, aqueous extraction and elution of active substance with 80% ethanol. Besides of heterophile antigen preparation obtained by the method of Schwerzweiss and Tomcsik (preparation S-T) two serologically++ active preparations were obtained (fraction I and IV), which ability to inhibit PBD agglutinating reaction and bovine red blood cells haemolysis was 16 and 8 times lower, respectively, than S-T preparation. The preparation of heterophile mononucleosis antigen obtained differed in latex coating efficacy. In order to prepare latex reagent MZ-I (from fraction I) a solution of preparation of 125 micrograms/ml concentration was used, for MZ-II (from fraction IV)--50 micrograms and for MZ-III (from preparation S-T)--15 micrograms/ml. The reagent MZ-I showed, the highest activity in agglutinating test with human serum containing heterophile mononucleosis antibodies while two others reacted with 2-4 times lover serum dilutions. Similar differentiated reactivity with these reagents was found in latex test with 15 sera from patients suspected of having infectious mononucleosis.     

Purification and characterization of a novel proteinase, chymopapain S

Chymopapain (EC 3.4.22.6) possesses an essential and a non-essential thiol group, but enzyme preparations produced hitherto always contained less than two thiol groups. Starting from commercial chymopapain or from papaya latex, we have prepared pure enzyme having two thiol groups, which is demanded by mechanistic investigations. The purification was performed by covalent chromatography on activated thiol-Sepharose column, which specifically binds thiol-containing proteins. Elution was effcted with cysteine in a stepwise manner, first at pH 5 then at pH 8. The pure enzyme had a molecular mass of24 700 as estimated by sodium dodecyl sulfate polyacrylamide gel-electrophoresis. Titration of the pure enzyme with 2,2'-dipyridyl disulfide at pH 9 exhibited a biphasic curve. This indicated that under the conditions employed the nonessential thiol group is more reactive than the essential one, which is a characteristic feature of chymopapain B. On the other hand, the magnitude of the rate constant of the same reaction at pH 4, as well as its pH-dependence, was characteristic of chymopapain A. The enzyme with the hybrid kinetic properties was denoted as chymopapain S. We conclude from the above findings that various forms of chymopapain can be classified by their reaction with 2,2'-dipyridyl disulfide.     

Flutter in flow-limited collapsible tubes: a mechanism for generation of wheezes

We studied flutter in collapsible tubes as a possible mechanism for the generation of respiratory wheezes. The pressure-flow relationships and the wall oscillations of thick-walled [wall thickness (h)-to-lumen radius (r) ratio 1:1.7 to 1.3] self-supporting latex and Silastic tubes mounted between rigid pipes were measured. A high-impedance vacuum pump was connected to the downstream end. Upstream and downstream valves were used to control corresponding resistances. We found loud honking sounds and tube wall oscillations that occurred only when the tubes were buckled and flow limiting, i.e., when the flow became constant and independent of downstream driving pressure. The overall range of oscillatory frequencies was 260-750 Hz for airflow, presenting as sharp peaks of power on the frequency spectrum. The oscillatory frequencies (f) were higher at higher fluid velocities (u) and with narrower distance between opposing flattened walls (2b), resulting from increasing downstream suction pressure and the transmural pressure becoming more negative. The effect of u and b on f for a latex tube (h-to-r ratio 1:1.7) were found to be f = 228 + 0.021 (u/b). These relationships were valid throughout the range of oscillations in this tube (283-720 Hz) and with flow rates of 12-64 l/min. The experimental data were compared with predictions of the fluid dynamic flutter theory and the vortex-induced wall vibrations mechanism. We conclude that viscid flutter in soft tubes is the more probable mechanism for the generation of oscillations in the soft tube model and is a possible mechanism for the generation of respiratory wheezes.