Successful Culture (successful + culture)

Distribution by Scientific Domains


Selected Abstracts


Methodology and Transport Medium for Collection of Helicobacter pylori on a String Test in Remote Locations

HELICOBACTER, Issue 6 2005
Helen M. Windsor
ABSTRACT Background.,Helicobacter pylori can be isolated from patients using the string test but contaminating oral and nasopharyngeal microflora need to be suppressed by rapid plating out onto selective culture media. Recently, use of this diagnostic method was enhanced by using a novel transport medium to collect specimens from subjects in a remote Australian clinic over 1300 km from the laboratory. Methods., Retrieved string tests were transported to the laboratory in chilled polystyrene boxes in 5 ml screw-cap bottles with 3 ml of a brain heart infusion broth plus antibiotics. These were 20 g/ml vancomycin, 10 g/ml trimethoprim, 10 g/ml cefsulodin, and 10 g/ml amphotericin B. A comparison was made between subjects who gargled with a chlorhexidine mouthwash before swallowing the string test and those who did not. Results., Forty-five urea breath test-positive subjects were tested and H. pylori was isolated from 34 of them. Successful culture was achieved from string tests that were in transit for up to 29 hours and where the maximum temperature in the transport box was 14 °C. The additional use of a mouthwash had a marked effect on the isolation rate. H. pylori was cultured from 75% of subjects who gargled but only from 39% who did not. Conclusions., This methodology and transport medium can broaden the use of the string test to more remote geographic areas where endoscopy is not feasible so that H. pylori isolates may still be obtained for diagnostic and epidemiologic studies. The value of this promising methodology of collection and transport should be assessed in a controlled study. [source]


Differentiation of human ameloblast-lineage cells in vitro

EUROPEAN JOURNAL OF ORAL SCIENCES, Issue 2006
Qiaomei Yan
Previous studies have shown that ameloblast-like cells can be selectively cultured from the enamel organ in a serum-free medium with low calcium concentrations. The purpose of this study was to further characterize this culture system to identify differentiated ameloblast-lineage cells. Tooth organs from 19,24-wk-old fetal cadavers were either frozen and cryosectioned for immunostaining, or digested in collagenase/dispase for cell culture. The cells were grown in keratinocyte media supplemented with 0.05 mM calcium, and characterized by morphology and immunofluorescence. Epithelial clones with two distinct morphologies, including smaller cobblestone-shaped cells and larger (5,15 times in size) rounded cells, began to form between day 8 and day 12 after culture. The cobblestone-shaped cells continued to proliferate in culture, while the larger cells proliferated slowly or not at all. These larger cells formed filopodia, usually had two or more nuclei and a radiating cytoplasm at the cell margin, and were more abundant with increasing time in culture. Both cell types stained for cytokeratin 14, and the larger cells appeared more differentiated, showing stronger staining for amelogenin and ameloblastin. Immunofluorescence of the tooth bud sections showed staining for these matrix proteins as ameloblasts differentiated from the inner enamel epithelium. These results show the successful culture of differentiating ameloblast-lineage cells, and lay a foundation for use of these cells to further understand ameloblast biology with application to tooth enamel tissue engineering. [source]


Discordant influence of amphotericin B on epirubicin cytotoxicity in primary hepatic malignant cells collected by a new primary culture technique

JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 2 2006
ZU-YAU LIN
Abstract Background:, The purpose of this prospective study was to investigate whether amphotericin B (AmB) had any potential role in the systemic chemotherapy of primary hepatic malignancy using cancer cells collected by the authors' method of primary culture. Methods:, The specimens obtained by ultrasound-guided fine-needle aspiration biopsy (22 G) from 15 patients with hepatocellular carcinoma (HCC) and one with cholangiocarcinoma were plated into culture flask without disaggregation by trypsin-ethylenediamine tetra-acetic acid solution. Six patients with HCC and one patient with cholangiocarcinoma (7/16, 44%) had successful culture and the cancer cells at the 4th passage were continuously exposed to therapeutic ranges of epirubicin (0, 0.5, 1.0, 1.5, 2.0 µg/mL) with or without the combination of 2.5 µg/mL AmB for 24 h. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was applied to evaluate the effects of the drugs. A human HCC cell line (HA 22T/VGH) was studied for comparison. Results:, Addition of AmB showed no influence on epirubicin cytotoxicity in two patients (one partial resistant HCC and one epirubicin-sensitive cholangiocarcinoma; 25%), augmentation of the epirubicin cytotoxicity in two patients (one total resistant HCC, partial resistant HA 22T/VGH cell line and one epirubicin-sensitive HCC; 37.5%), and decrease of epirubicin cytotoxicity in the remaining three (one partial resistant and two epirubicin-sensitive HCC; 37.5%). Conclusions:, Amphotericin B has a discordant influence on epirubicin cytotoxicity in primary cultured hepatic malignant cells. Application of AmB in the systemic chemotherapy of primary hepatic malignancy should be limited to patients with positive AmB effect evaluated by an in vitro sensitivity test such as the present method. [source]


A multi-institutional studyof orthotopic neobladders: functional results in men and women

BJU INTERNATIONAL, Issue 6 2004
R. Carrion
Multi-institutional studies are extremely valuable whena new surgical technique is being introduced. The Confederationof American Urology conducted such a study into the functional results inmales and females of the orthotopic bladder. In a series of 138patients they found many interesting outcomes, which they presenthere. Authors from Sheffield describe their experience in developing tissue-engineeredbuccal mucosa for use in urethral reconstruction. They describetheir technique and report the successful culture of full-thicknessbuccal mucosa, which they found to be robust and safe for clinicaluse. OBJECTIVES To analyse the incidence of diurnal incontinence (DI) and nocturnalincontinence (NI), the need for intermittent catheterization (IC),and the rate of ureteric obstruction (UO) among a group of men andwomen with ileal and colonic orthotopic neobladders in four countries. PATIENTS AND METHODS In all, 138 patients (113 men and 25 women) had an orthotopicneobladder constructed after radical cystectomy for carcinoma. The mean(range) age was 61.3 (28,76) years and thefollow-up 41 (6,144) months. All patients underwentsurgery by experienced surgeons associated with the Confederationof American Urology. A retrospective evaluation was designed toreview the functional results and the incidence of UO. The techniqueof orthotopic neobladder construction was at each surgeon's discretion. Various detubularized bowelsegments were used, including ileum, colon or sigmoid. Patients werefollowed by chart reviews and personal interviews at 1, 3 and 6 monthsafter surgery and then every 6 months, and were evaluated bya physical examination, urine analysis, cytology and renal ultrasonography. RESULTS An ileal or colonic neobladder was constructed in 74 and 64 patients, respectively. Five (7%), 23 (31%), 10 (14%) and 14(9.6%) with an ileal neobladder developed DI, NI, IC andUO, respectively; the respective values for patients with a colonicneobladder were eight (12%), 19 (30%), seven (11%)and 15 (12%). Statistical analysis by Fisher'sexact test showed no significant differences between the ileal andcolonic neobladder groups or with gender. CONCLUSIONS Using this specific protocol for evaluating many men and womenwith ileal and colonic orthotopic neobladders showed no significant differencesin the incidence of DI, NI, IC or UO. Neobladders constructed fromdetubularized bowel, irrespective of bowel segment(s) used, canprovide satisfactory diurnal results. A moderate incidence of NIand UO continue to be a problem. [source]