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Urinary Tract Disease
Published in Vincenzo Berghella, Maternal-Fetal Evidence Based Guidelines, 2022
A threshold of ≥100,000 colony-forming units (CFUs) per milliliter of the same bacterial strain on two consecutive voided specimens is the indication for treatment [48]. The detection of ≥100,000 CFU/mL in a single voided midstream urine sample is accepted as an adequate and more practical alternative, although there is only an 80% probability the woman has true bacteriuria; this increases to 95% if two or more consecutive cultures are positive for the same organism [59]. Additionally, asymptomatic bacteriuria may be defined as >100,000 CFU/ml of a single recognized uropathogen when the specimen was obtained with a catheterized specimen [48]. Because the performance of rapid urine screening tests in pregnancy is poor, quantitative culture remains the gold standard for diagnosis. Group B Streptococcus should be appropriately treated at any concentration (see Chap. 18, Obstetric Evidence Based Guidelines). It is important to avoid contamination by cleansing the perineum and then collecting “mid-stream” urine.
Antimicrobial Preservative Efficacy and Microbial Content Testing*
Published in Philip A. Geis, Cosmetic Microbiology, 2020
Scott V.W. Sutton, Philip A. Geis
Freshly grown stocks of a particular culture are prepared by inoculating a solid agar medium. Incubate bacterial cultures at 30–35°C for 18–24 hours. Incubate yeasts at 20–25°C for 44–52 hours, and molds at 20–25°C for 6–10 days. Then harvest the bacteria and yeast using sterile saline (0.9% NaCl) and dilute the suspended cells to 1 × 108 CFU/mL. The mold is harvested with sterile saline containing 0.05% Polysorbate 80, adjusting the spore count to 1 × 108 CFU/mL. The number of colony forming units per milliliter determines the amount of inoculum to use in the test. The viability of the suspension should be monitored, especially if not used promptly.
Bone Marrow Harvesting and Reinfusion
Published in Adrian P. Gee, BONE MARROW PROCESSING and PURGING, 2020
Work with mice has shown that spleen colony-forming units (CFU-S) tolerate freezing very well,23 but human bone marrow is more difficult to evaluate. Attempts have been made to quantify human marrow cell viability following freezing, by assessing in vitro colony-forming ability,24,25 and by trying to correlate the recovery of colony-forming units-granulocyte-macrophage (CFU-GM) from frozen marrow with the subsequent rate of engraftment.26 A fuller account of the early history of bone marrow cryopreservation is given by Pegg,27 and modern techniques are described by Law and Meryman (Chapter 22) and by Stiff (Chapter 23).
Influence of biofilm removal from the tooth-restoration interface on the progression of secondary caries lesions: a preliminary in vitro model study
Published in Biofouling, 2020
Cácia Signori, Tamires Timm Maske, Vitor Henrique Digmayer Romero, Maximiliano Sérgio Cenci
The specimens were individually removed from the wells and carefully washed in sterile saline. The biofilm was collected from the surface (enamel/gap/resin) with sterile plastic instruments and deposited in pre-weighed microtubes. The contents of the tubes were vortexed and sonicated (Sonicador Vibra Cell - Sonics and Materials, Danbury, CT, USA) at a power of 40W and amplitude of 5% using 6 pulses of 9.9s each. Thereafter, the suspensions were diluted in saline (10−1:10−7) and inoculated in duplicate in the culture media, including blood agar for total microorganism quantification, mitis salivarius agar with 0.2 units of bacitracin ml−1 for the visual assessment of mutans streptococci, brain heart infusion agar (pH 4.8) for acid-tolerant microorganisms, and Rogosa agar for lactobacilli. The plates were incubated under an anaerobic atmosphere at 37°C for 96h. After, the numbers of colony-forming units (CFUs) were then determined by one trained operator based on colony morphology and cell morphology using a microscope and stereoscopic microscope. Representative colonies of mutans streptococci were confirmed by their capacity to ferment mannitol, sorbitol, melibiose and raffinose (Shklair and Keene 1974). Microbial counts were expressed as CFU mg−1 biofilm (wet weight).
Antimetabolic Agent 3-Bromopyruvate Exerts Myelopotentiating Action in a Murine Host Bearing a Progressively Growing Ascitic Thymoma
Published in Immunological Investigations, 2020
Saveg Yadav, Shrish Kumar Pandey, Yugal Goel, Mithlesh Kumar Temre, Sukh Mahendra Singh
Bone marrow colonies were prepared according to a method described earlier using culture medium containing methylcellulose (Vishvakarma and Singh, 2011b). Briefly, BMC (1 × 104 cells/ml) were suspended in a mixture containing 0.9% (w/v) methylcellulose with 30% (v/v) FCS and 20% (v/v) L929CM. The mixture was gently vortexed, plated in a 35-mm plastic culture dish (Greiner Diagnostic GmbH, Germany) and incubated at 37°C in a humidified atmosphere of 5% CO2 in air. Bone marrow colonies were counted after 10 days of incubation. An aggregate of more than 25 cells was counted as a single colony-forming unit (CFU). Colonies of different types were identified based on their morphological features. Colonies with macrophage-like morphology were designated as CFU-M, granulocyte–macrophage like morphology as CFU-GM and granulocyte morphology as CFU-G.
Novel cystamine-core dendrimer-formulation rescues ΔF508-CFTR and inhibits Pseudomonas aeruginosa infection by augmenting autophagy
Published in Expert Opinion on Drug Delivery, 2019
Janine Faraj, Manish Bodas, Garrett Pehote, Doug Swanson, Ajit Sharma, Neeraj Vij
CF epithelial cells are known to suffer from impaired bacterial phagocytosis secondary to mutant ΔF508-CFTR accumulation, which is restored by autophagy induction [22,26]. To assess the degree of phagocytosis restoration induced by our novel dendrimer formulation, we used CFBE41o-cells incubated with G4-DAB, G4-CYS and an untreated group serving as a control, followed by infection of the cells with PA01-GFP. Microscopic analysis and quantification revealed significantly higher percentages of PA01-infected cells in both G4-DAB (*p < 0.05) and G4-CYS (**p < 0.01) treatment groups (Figure 5(a,b)). Colony forming units (CFUs) were also measured as an indicator of bacterial survival. Our data reveal a significant reduction in bacterial survival with treatment of G4-CYS (*p < 0.05, Figure 5(c)); although decreased bacterial survival was observed with G4-DAB treatment, measurements were determined to be statistically insignificant. Our data confirm that these novel G4-DAB and G4-CYS dendrimers are capable of restoring autophagic and phagocytic processes in CF epithelial cells.