Evaluation of a rapid bacteriuria screening instrument.

A newly modified, semiautomated instrument (Bacteriuria detection device (BDD] designed to detect the presence of bacteriuria in less than 3 min was compared to quantitative urine culture plating techniques. The instrument consists of a self-enclosed vacuum-filtration-staining system in which a 1-ml urine filtrate is stained on a filter. The resulting color determines the quantitation. Of the total of 525 clinical urine specimens tested, 66 (12.6%) were uninterpretable due to pigment deposition or inability to complete the filtering process (clogging of the filter). Of the remaining 459 specimens, 93 (20.3%) had a plate quantitation colony count of 10(5) colony-forming units (CFU)/ml or more. The BDD detected 94.2% of these positive specimens if only significant pathogens were included (85% if specimens that were probably contaminated were also included). For specimens containing significant pathogens at 10(4)-10(5) CFU/ml, the BDD detection rate was 41%. The device detected most (94.3%) gram-negative bacilli and enterococci at colony counts of 10(5) CFU/ml or more. In addition, the BDD accurately detected 95.6% of specimens with no growth or fewer than 10(4) CFU/ml. With several proposed modifications, these results suggest that this instrument is potentially useful as a urine screening device in a select population.

Investigation of polymeric nanoparticles as carriers of enalaprilat for oral administration.

Enalaprilat is a typical angiotensin-converting enzyme inhibitor and is very poorly absorbed from the gastrointestinal tract. The aim of this study was to design and characterize poly-(lactide-co-glycolide) (PLGA) and polymethylmethacrylate (PMMA) nanoparticles containing enalaprilat and to evaluate the potential of these colloidal carriers for the transport of drugs through the intestinal mucosa. Nanoparticle dispersions were prepared by the emulsification-diffusion method and characterized according to particle size, zeta potential, entrapment efficiency and physical stability. Effective permeabilities through rat jejunum of enalaprilat in solution and in enalaprilat-loaded nanoparticles were compared using side-by-side diffusion chambers. The solubility of enalaprilat is very low in many acceptable organic solvents, but in benzyl alcohol is sufficient to enable the production of nanoparticles by the emulsification-diffusion process. The diameters of drug-loaded PMMA and PLGA nanoparticles were 297 and 204 nm, respectively. The concentration of the stabilizer polyvinyl alcohol (PVA) in dispersion has an influence on particle size but not on drug entrapment. The type of polymer has a decisive influence on drug content--7 and 13% for PMMA and PLGA nanoparticles, respectively. In vitro release studies show a biphasic release of enalaprilat from nanoparticle dispersions-fast in the first step and very slow in the second. The apparent permeability coefficient across rat jejunum of enalaprilat entrapped in PLGA nanoparticles is not significantly improved compared with enalaprilat in solution.

Interactions of solid lipid nanoparticles with model membranes and leukocytes studied by EPR.

Solid lipid nanoparticles (SLN) are colloidal systems which have been proposed for several administration routes. Only limited data are available about the mechanism and rate of interaction of SLN with cells and tissues. The aim of our study was to investigate interactions of SLN with model membranes (liposomes) and cells (leukocytes). SLN dispersions composed of glyceryl tripalmitate, phosphatidylcholine, water, and poloxamer 188 or Tween 20 were prepared by the melt-emulsification process. Spin-labeled phosphatidylcholine (PC(10,3)) and the methylester of doxyl palmitic acid (MeFASL(10,3)) were incorporated into SLN as spin probes (SPs) in order to determine the rate and mechanism of cell interaction by electron paramagnetic resonance (EPR) spectroscopy. Our results indicate that the exchange of SP between SLN and liposomes is much faster for MeFASL(10,3) than for PC(10,3), probably due to the smaller size of the former. In contrast to liposomes, in leukocytes no significant difference in the transfer rates of the two SP was observed after incubation, suggesting that there is an uptake of SLN to leukocytes (endocytosis) although simultaneous SP diffusion is not excluded. The interaction of SLN with leukocytes appears to depend significantly on the stabilizer used. Transfer of PC(10,3) from SLN coated with poloxamer 188 is much faster than from SLN coated with Tween 20.

Influence of spin probe structure on its distribution in SLN dispersions.

Solid lipid nanoparticles (SLN) are drug carrier system composed of biodegradable substances, which are solid at room temperature. The physico-chemical properties and structure of the incorporated compounds can affect their partitioning in SLN dispersions. In this work the influence of lipophilicity and structure of different SP on its location in SLN were studied. By electron paramagnetic resonance (EPR) measurements it was found that lipophilic SP distribute between a solid glyceride core and a soft phospholipid layer, with the more polar part (piperidine ring or methylcarboxylic groups) oriented toward the water-lipid interface. The majority of SP is located in the phospholipid layer, but the portion in the solid lipid core increases with SP lipophilicity. The hydrophilic Tempol does not incorporate into SLN.

Effect of urine preservation on urine screening and organism identification.

Three urine preservation-transport methods were examined for their effect on rapid urine-screening procedures. Results from fresh urine specimens, screened for bacteriuria by leukocyte esterase, nitrate, Autobac, Bac-T-Screen, Auto Microbic System (AMS), and bioluminescence procedures, were compared with urine-screen results from urine specimens held for 24 hours at room temperature in chemical preservatives. Quantitative discrepancies ranged from 0%, for urine preserved with glycerin-boric acid-sodium formate and tested by AMS or leukocyte esterase, to 21% for urines in the same preservative tested by bioluminescence or nitrate. Up to 62% of the organism identifications made from preserved urine specimens tested by the AMS urine card were in error. These data suggest that it may be inadvisable to use weak organic acid-based urine preservation systems in conjunction with these urine-screen procedures.

Differentiation of Haemophilus spp. in Respiratory isolate cultures by an indole spot test.

Indole spot tests using isolated, nonhemolytic colonies of Haemophilus species were positive for 90 of 151 (60%) respiratory isolates of Haemophilus influenzae, whereas 67 to 72 (93%) isolates of H. influenzae from cerebrospinal fluid and blood specimens were indole positive. Only 4 of 117 (3%) Haemophilus parainfluenzae isolates were positive for indole spot tests. Thus, indole-positive, nonhemolytic Haemophilus isolates in respiratory cultures can be presumptively identified as H. influenzae.

Transition in quality: from quality assurance to strategic quality management.

The 1990s and beyond present formidable challenges to health-care providers, including clinical laboratories and pathology departments. However, numerous opportunities lie within these challenges. Discovering these opportunities and exploiting them will be critical success factors for future survival. Quality assurance, continuous quality improvement, and strategic and financial planning are all activities used to a varying extent by clinical laboratories. The cumulative potential benefits to an organization in which these activities are integrated can far exceed their sum as individual components. Coordinating these interdependent processes is the basis for managing strategically. The experience of one organization's efforts to plan and develop such a strategy is presented and discussed.

Bacterial flora of the vagina during the menstrual cycle: findings in users of tampons, napkins, and sea sponges.

The recent association of menstruation, tampon use, and staphylococcal infection with toxic shock syndrome led us to study the association of menstruation and catamenial product use with changes in vaginal flora. Cultures of the cervical os were obtained in midcycle and during menstruation from 12 women who used napkins and 40 women who used tampons. Staphylococcus aureus was found during midcycle and menstruation in three women, during menstruation alone in six, and during midcycle alone in none, indicating a significant association of S. aureus with menstruation (p = 0.04). No difference was found in the rate of S. aureus colonization during menstruation in tampon users (18%) and napkin users (17%). In a similar study, cultures were taken for S. aureus and other aerobic bacteria from 58 tampon users and 25 users of sea sponges. Staphylococcal colonization was found to be increased during menstruation in both groups. Among the cultures done during menstruation, those from users of sea sponges were found to have significantly higher colonization rates with S. aureus, Escherichia coli and other Enterobacteriaceae. The association of sea sponges with a high rate of S. aureus colonization suggests that they are not an alternative to tampons for women seeking to decrease the risk of toxic shock syndrome.

Selective and differential medium for recovery of Pseudomonas cepacia from the respiratory tracts of patients with cystic fibrosis.

A selective and differential medium, OFPBL (oxidation-fermentation base supplemented with agar, lactose, and two antimicrobial agents), for the isolation of Pseudomonas cepacia from respiratory specimens of patients with cystic fibrosis was developed and tested. Among 725 specimens submitted from seven centers over a 4- to 6-month period, 58 (8%) yielded P. cepacia on OFPBL; only 19 of these were recovered on MacConkey or sheep blood agar (P less than 0.001). No isolate was recovered on MacConkey or sheep blood agar alone. Ranges of recovery rates among centers were 0 to 15% on OFPBL and 0 to 10% on MacConkey or sheep blood agar. Ninety percent of P. cepacia isolates were detected on OFPBL in less than or equal to 3 days. Other nonfermenters and yeasts isolated on OFPBL were distinguished from P. cepacia by failure to acidify the medium. The new medium was clearly superior to MacConkey and sheep blood agars for the isolation of P. cepacia from the respiratory secretions of patients with cystic fibrosis.