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Biomedical subjects

A W Bruce

Publications and source records attributed to A W Bruce.

At least 19 recordsLinked to original sources

Correlation between hydrophobicity and resistance to nonoxynol-9 and vancomycin for urogenital isolates of lactobacilli.

Seven clinical isolates of lactobacilli were found to be relatively hydrophobic with a mean water-contact angle of 66 +/- 15 degrees, and to be susceptible to 1% nonoxynol-9 and vancomycin. However, seven other strains were relatively hydrophilic with a mean water-contact angle of 32 +/- 13 degrees, and found to be resistant to 25% nonoxynol-9 and vancomycin. Thus, the surface properties of lactobacilli that influence susceptibility to antimicrobial agents may involve surface hydrophobicity. Possibly the penetration barrier posed by the cell surface towards these two nonionic antimicrobials is lower for hydrophobic cells than for hydrophilic cells.

Adult

Antibiotic resistance of urinary pathogens isolated from patients attending the Toronto Hospital between 1986 and 1990.

A study was carried out on 1523 urinary isolates obtained at The Toronto Hospital, Canada's largest tertiary care establishment, over three 1-month periods in 1986, 1987 and 1990. Escherichia coli was the most frequently isolated organism, with Enterococcus spp. the second most common isolate in 1986 and 1987, and Streptococcus spp. in 1990. Pseudomonas aeruginosa isolates were found to be resistant to many of the antimicrobial agents tested. Resistance patterns were found to commonly prescribed ampicillin, co-trimoxazole and, to some extent, the new fluoroquinolones, ciprofloxacin and norfloxacin. These results are relevant to the treatment and management of urinary tract infections in patients attending a tertiary care hospital.

Cross Infection

Hydrogen peroxide production by Lactobacillus species: correlation with susceptibility to the spermicidal compound nonoxynol-9.

Facultative anaerobic lactobacilli were recovered from the vaginas of 96.8% of 63 nonpregnant, healthy, premenopausal women. The predominant species were Lactobacillus jensenii, Lactobacillus acidophilus, and Lactobacillus casei. Of the women, 74.6% had hydrogen peroxide-producing lactobacilli, 22.2% had non-hydrogen peroxide-producing lactobacilli, and 3.2% had no lactobacilli. None of the 68 isolates had catalase activity. Some 68.2% of the isolates were inhibited by concentrations of less than or equal to 1% (wt/vol) of nonoxynol-9 (bactericidal for 73.3% of isolates, bacteriostatic for 26.7%). The remaining 31.8% could grow in all concentrations to 25% (wt/vol) of nonoxynol-9. All of the lactobacilli that were sensitive to nonoxynol-9 produced hydrogen peroxide whereas only 3 of 21 resistant strains were hydrogen peroxide producers. A significant correlation (P less than .001, chi 2 test) was found between hydrogen peroxide production and sensitivity to nonoxynol-9. It is suggested that the vaginal flora of spermicide users could be depleted of hydrogen peroxide-producing lactobacilli, possibly increasing susceptibility to urogenital infection.

Adult

Comparison of contact angles and adhesion to hexadecane of urogenital, dairy, and poultry lactobacilli: effect of serial culture passages.

The aim of this study was to examine the hydrophobicities of 23 urogenital, dairy, poultry, and American Type Culture Collection isolates of lactobacilli and to determine the effect on hydrophobicity of serially passaging the strains in liquid medium. To this end, strains were grown after isolation and identification and then serially passaged up to 20 times. Hydrophobicity was assessed through contact angle measurements on lawns of cells by using water, formamide, methylene iodide, 1-bromonaphthalene, and hexadecane as wetting agents and through measurement of their partitioning in a hexadecane-water system. The hydrophobicities of these strains varied widely, with Lactobacillus casei strains being predominantly hydrophilic and L. acidophilus strains being mostly hydrophobic. For some isolates, serial passaging was accompanied by a clear loss of hydrophobic surface properties, whereas for other strains, cultures became heterogeneous in that some cells had already lost their hydrophobic surface properties while others were still hydrophobic. Adhesion of this collection of lactobacilli to hexadecane droplets in microbial adhesion to hexadecane (MATH) tests was driven by their aversion to water rather than by their affinity for hexadecane, as concluded from the fact that hexadecane contact angles were zero for all strains. Furthermore, adhesion of the lactobacilli to hexadecane in MATH tests occurred only when the water contact angle on the cells was above 60 degrees.

Adult

Ability of uropathogens to bind to Tamm Horsfall protein-coated renal tubular cells.

Epithelial cells were isolated from a healthy human kidney and found to be of normal, renal tubular origin. The cells were maintained in tissue culture and found to secrete Tamm Horsfall protein (THP). Three strains of uropathogenic Escherichia coli, and one each of Proteus mirabilis and Pseudomonas aeruginosa were found to adhere to the THP-coated tubular cells. Extraneous THP bound to all the organisms except P fimbriated E. coli, and caused a reduction in the adhesion of type 1 fimbriated E. coli and P. mirabilis to the renal cells. However, irrespective of reduced adhesion in three of five strains tested, there was adequate evidence to indicate that THP does not competitively exclude all uropathogenic adhesion. On the contrary, its presence on renal cells may act to mediate bacterial colonization.

Bacterial Adhesion

Effect of extracorporeal shock wave lithotripsy on bacterial viability. Relationship to the treatment of struvite stones.

The aim of this study was to determine whether extracorporeal shock wave lithotripsy (ESWL) affected the viability of the infecting bacteria within a simulated struvite stone matrix. A strain, Proteus mirabilis 28cii, was prepared in three forms: (1) suspended in saline and urine, (2) artificially encapsulated by suspending in agar beads and (3) artificially encapsulated and mineralised by suspending in agar beads with calcium carbonate crystals. The preparations were placed in capped vials partially immersed in degassed water and held in the focal point of the Siemens Lithostar and given 1,000 shocks. Subsequent viability testing showed that bacteria suspended in urine were greatly affected by shock treatments (55% loss in viability), but incorporation into agar beads negated this effect (even if the cells were exposed to 2000 shocks). Mineralisation of the beads with calcium carbonate crystals caused a decrease in viability of 82% that was significantly different from controls. However, this still left 2.3 X 10(8) viable organisms (82% of 2.8 X 10(8], easily enough to form the focus for further infections. A series of control experiments carried out using an ultrasonic cell sonicator probe gave comparable results to those obtained with ESWL. These results demonstrate the ESWL treatment of infected stones must be accompanied by antimicrobial coverage.

Humans

Is there a role for lactobacilli in prevention of urogenital and intestinal infections?

This review describes the importance of microbial adhesion in the ecology of the urogenital and intestinal tracts and the influence of host and microbial factors in bacterial interference. In a recent revival of interest in bacterial interference, lactobacillus administration has been studied as a means of treating and preventing disease. Although evidence is conflicting, Lactobacillus acidophilus appears to be involved in beneficial antagonistic and cooperative reactions that interfere with establishment of pathogens in the gastrointestinal tract. The mechanisms of action are believed to involve competitive exclusion and production of inhibitory substances, including bacteriocins. These characteristics, as well as demonstrated adherence abilities in vitro, led to selection of certain Lactobacillus strains for clinical studies of cystitis. Weekly intravaginal Lactobacillus therapy reduced the recurrence rate of uncomplicated lower urinary tract infections in women. Use of Lactobacillus strains resistant to Nonoxynol-9, a spermicide that kills members of the protective normal vaginal flora, may have potential for use in women with recurrent cystitis using this contraceptive agent. In veterinary studies, bacterial interference by administration of probiotics has also been beneficial in disease prevention in animals. Carefully selected bacterial mixtures integrate with the gastrointestinal flora of the animals and can confer disease resistance and improve physiological function. Additional human and animal trials are needed to determine the practical, long-term usefulness of bacterial interference as a protective mechanism against infectious diseases.

Bacterial Physiological Phenomena

The spermicidal compound nonoxynol-9 increases adhesion of Candida species to human epithelial cells in vitro.

All 25 strains of Candida spp. tested were able to grow in medium supplemented with 25% nonoxynol-9 in vitro. Adhesion of Candida spp. to HeLa cells was found to increase in the presence of 5% nonoxynol-9 (2.2- to 6.6-fold; P less than 0.001) and, to a lesser extent, in 12.5% nonoxynol-9. Adhesion of Candida strains cultured in medium supplemented with nonoxynol-9 varied, with five of six strains of Candida albicans and the single strain of Candida tropicalis demonstrating increases of 1.4 to 4.0 times control levels (P less than 0.05). The increased adhesion did not correlate with altered yeast cell surface hydrophobicity or germ tube formation.

Adhesiveness

Effect on urogenital flora of antibiotic therapy for urinary tract infection.

Specimens were examined from 70 female presenting with urinary tract infection. 50 patients were randomly treated with 7 days amoxicillin or bacampicillin and another 20 randomly received amoxicillin or enoxacin. Effective clearance of the infections was achieved with each antibiotic, and only minor side effects occurred. Bacteriological analyses of the urogenital flora demonstrated a relationship between urethral carriage and bladder infection, with Escherichia coli being the most common pathogen. Antibiotic resistant E. coli were isolated more frequently from the bladder, urethra, introitus and rectum following amoxicillin treatment compared to bacampicillin and enoxacin. An examination of the urogenital flora post therapy showed that an indigenous lactobacillus population had not been restored in the majority of patients. Rather, uropathogenic bacteria were found to dominate the urethra and introitus. The effects of antibiotics on urogenital flora is clearly a matter of importance. Future clinical trials should address this. It may be that artificial supplementation of indigenous bacteria is necessary to restore the flora back to normality.

Adolescent

Bacterial aggregation in relation to peritoneal dialysis.

The ability of gram-positive cocci to aggregate and form biofilms at the exit site and on catheter surfaces in continuous ambulatory peritoneal dialysis (CAPD) has been associated with peritonitis. In the present study, 16 (80%) of 20 dialysate effluents from patients with staphylococcal peritonitis were found to contain aggregates of Staphylococcus epidermidis or S. aureus. Using an in vitro assay system, 20 (83%) of 24 pathogenic bacterial strains were found to aggregate in fresh dialysis solutions within 4 to 24 h at 37 degrees C. Further studies with one strain, S. epidermidis 1938, showed increased aggregation when the bacteria were incubated in dialysate effluents compared to fresh dialysis solutions. Viability experiments showed that aggregated bacteria were less sensitive to the killing effects of fresh dialysis solutions and that dialysate effluents could support bacterial growth. The results suggest that attention be paid to aggregation as a potential mechanism used by bacteria in causation of peritonitis.

Bacterial Adhesion

Prostatitis associated with Chlamydia trachomatis in 6 patients.

A number of studies have shown that Chlamydia trachomatis bacteria are important pathogens in urogenital tract infections, such as acute urethritis, acute epididymo-orchitis in male patients and pelvic inflammatory disease in female patients. We confirm a previously reported association between Chlamydia trachomatis and prostatitis. We report on 6 patients with definitive confirmation via tissue culture and an immunofluorescent technique. The results demonstrate that chlamydial prostatitis occurs in patients with nonbacterial disease. Efforts should be made to detect and eradicate the organisms from the prostate.

Adult

Effect of antibiotic treatment on receptivity of uroepithelial cells to uropathogens.

The management of female patients with recurrent urinary tract infections still remains a problem, and long-term prophylactic or short intermittent courses of antibiotics are the standard forms of therapy. In this report, 10 patients were examined for the effects of long- and short-term treatment with trimethoprim-sulfamethoxazole (TMP-SMX) antibiotics on the receptivity of uroepithelial cells to bacterial adherence. The urine of all patients was sterile while on antibiotic therapy. Few bacteria were found adherent to the cells from adult patients (group 1, mean age 36 years) on long-term antibiotics, but the cells were highly receptive to uropathogens in vitro, especially for Escherichia coli expressing mannose-resistant adhesins. Controls of age-matched adult females were included and in vitro adherence levels were found to be higher for those women with a history of urinary tract infection compared with those with no past record of infection. In the second group, elderly patients (mean age 87 years) presented with bacteriuria, and their uroepithelial cells were found to be colonized by uropathogens to a significantly greater extent than their controls. The adherent population was reduced during 7-day TMP-SMX antibiotic treatment, but increased posttherapy, particularly in two patients who subsequently became reinfected. The in vitro results showed that uroepithelial cells retain their receptivity to uropathogenic adherence, both during and after treatment. Although antibiotics eradicate uropathogens from the urinary tract, patients remain susceptible to recolonization by uropathogens and are at risk of reinfection after completion of therapy.

Adult

Intravaginal instillation of lactobacilli for prevention of recurrent urinary tract infections.

Urinary tract infections remain a common problem, particularly in the female population. New methods are required to manage recurrent cystitis, and extensive research to date has suggested that restoration of the lactobacilli flora of the urogenital tract may prevent these infections. In this study, five females suffering from recurrent urinary tract infections were treated twice weekly with intravaginal and perineal implantation of Lactobacillus casei GR-1. These organisms colonized the epithelium and prevented the emergence of coliform bacteria in most instances, but did not appear to affect enterococcal colonization. In vitro studies showed that L. casei GR-1 inhibited the growth of the coliforms but did not inhibit enterococci. Each of the five patients had infection-free periods ranging from 4 weeks to 6 months. The treatment was well tolerated, had no side effects, led to an improved well-being, and was preferred to antibiotic treatment by all of the patients. These human studies, albeit of a limited nature, are the first to examine the potential for lactobacillus therapy in the prevention of urinary tract infections. The results show that lactobacilli therapy, using carefully selected organisms to treat patients who are closely followed, may be effective in the prevention of recurrent urinary tract infections.

Adolescent

Examination of strains of lactobacilli for properties that may influence bacterial interference in the urinary tract.

Lactobacilli strains vary in their ability to interfere with colonization of uroepithelial cells by pathogenic organisms. Bacterial size, adherence capability, competitive exclusion and inhibition of pathogenic growth were examined in selected strains of lactobacilli to assess their relative importance. We tested adherence to uroepithelial cells of 34 lactobacillus isolates originating from the urogenital tract, dairy sources and chickens, and 1 lactobacillus strain isolated from a commercially available preparation. The ability to adhere differed greatly among the 34 strains, with a range of 8 to 74 bacteria per cell, and the commercial strain attached poorly with a mean of 15 bacteria per cell. Eleven strains of lactobacilli were selected and found to differ in length and width dimensions. These strains were found to exclude competitively with varying efficiency 6 strains of uropathogens from adhering to uroepithelial cells. The effective ability of Lactobacillus casei GR-1 to exclude the uropathogens confirmed previous findings and other strains (GR-2, GR-3 and A-60) gave a similar range of exclusion. The lactobacillus isolates varied greatly in their ability to inhibit the growth of 11 uropathogens under the conditions of an agar overlay inhibition assay. A combination score was allocated to each of the 11 lactobacillus strains based on size, adherence, exclusion and inhibition. Lactobacillus casei GR-1 attained the highest score while the commercial strain attained the lowest score. The results of this study identified 4 properties that should be examined to select lactobacillus strains before embarking upon bacterial interference studies to prevent urinary tract and other infections in humans.

Animals

Ultrastructural microbiology of infected urinary stone.

Scanning and transmission electron microscopy of an antibiotic-treated infected urinary stone demonstrates a thick bacterial biofilm adhering to the surface of the stone. Cultures of the crushed calculi indicated the viability of the bacteria. Exopolysaccharide secreted by the bacteria develops a matrix or glycocalyx that appears to protect the biofilm-forming sessile bacteria from antibiotics.

Aged

Production of monoclonal antibodies against prostatic acid phosphatase by in vitro immunization of human spleen cells.

Monoclonal antibodies against human prostatic acid phosphatase (PAPase) were produced by immunization of human primary spleen cell cultures. Dissociated spleen cells were cultured for 5-8 days in the presence of 100 ng/ml of PAPase and pokeweed mitogen (1:5000). Following immunization, B cells were isolated and infected with Epstein-Barr virus (EBV). Two weeks after EBV-transformation, cells were fused with either mouse myeloma cells (SP2/OAg14) or human/mouse heteromyeloma cells (SHM-D33). Hybrid clones were screened for anti-PAPase production. In 7 independent immunizations, the average fusion frequency was 3.6 per 10(6) lymphocytes. 18-32% of the hybridomas produced anti-PAPase; approximately 75% of these secreted IgM and 25% secreted IgG. Antibody specificity was determined by immunoassay and immunohistological studies. The procedures described here may be suitable for the production of human monoclonal antibody of a useful specificity.

Acid Phosphatase

Competitive exclusion of uropathogens from human uroepithelial cells by Lactobacillus whole cells and cell wall fragments.

Previous studies have shown that indigenous bacteria isolated from cervical, vaginal, and urethral surfaces of healthy women are able to adhere to human uroepithelial cells in vitro. Furthermore, these organisms were found to block the adherence of uropathogenic bacteria to uroepithelial cells from women with and without a history of urinary tract infections. In the present study, complete or partial inhibition of the adherence of gram-negative uropathogens was achieved by preincubating the uroepithelial cells with bacterial cell wall fragments isolated from a Lactobacillus strain. Competitive exclusion was most effective with whole viable cells and less effective with cell wall fragments obtained by sonication, extraction with sodium dodecyl sulfate, and treatment with sodium dodecyl sulfate and acid. Analysis of the Lactobacillus cell wall preparations suggested that lipoteichoic acid was responsible for the adherence of the Lactobacillus cells to uroepithelial cells but that steric hindrance was the major factor in preventing the adherence of uropathogens. This conclusion was also supported by blockage studies with reconstituted lipoteichoic acid-peptidoglycan, which was more effective at blocking adherence than lipoteichoic acid or peptidoglycan alone. The results suggest that the normal flora of the urinary tract may be used to protect against the attachment of uropathogens to the surfaces of uroepithelial cells. The long-term implications of these findings may lead to alternative methods for the management and prevention of recurrent urinary tract infections in females.

Adult