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At least 19 recordsLinked to original sources

Isolation of organisms in CAPD peritonitis: use of nutrient broth cultures and Bactec blood culture media.

Cultures of the first cloudy dialysates from 51 consecutive episodes of peritonitis in patients undergoing continuous ambulatory peritoneal dialysis were carried out using concentrated nutrient broth and Bactec blood culture media in addition to primary culture on solid media. Thirty-seven episodes (73%) were positive on solid media, and 42 episodes (82%) were positive by both nutrient broth and Bactec methods. The value of Gram-stained smears and WBC counts on dialysates is discussed.

Bacteria↗

Generation of dendritic cells from human peripheral blood monocytes--comparison of different culture media.

Culture medium or medium supplement is one of the factors responsible for dendritic cell (DC) generation, but little is known about the influence of various media on DC culture. In our study we generated DC from adherent monocytes of human peripheral blood in the presence of GM-CSF, IL-4 and TNF-alpha. The following culture media were used: RPMI 1640 supplemented with 2% human serum albumin; RPMI 1640 supplemented with 2% TCH serum replacement; X-VIVO 15 and Panserin 501. Flow cytometry analysis revealed that in all media cells were CD83+ and lost CD14. Interestingly, the use of Panserin and RPMI with albumin preferentially gave rise to CD1a+ DC, whereas in X-VIVO and RPMI with TCH we observed both CD1a+ and CD1a-. Our results showed that RPMI with TCH yielded the highest percentage of cells expressing both CD80 and CD86 molecules and, in contrast to other media, the higher percentage of CD86+ cells in comparison to CD80+ cells.

Antigens, CD↗

Prediction of mecA-positive coagulase-negative staphylococci: assessment of different phenotypic methods, breakpoints, culture media and culture conditions.

Sensitivity for the detection of mecA-positive coagulase-negative staphylococci (CNS) was evaluated for different breakpoints of resistance for oxacillin using three different susceptibility tests, either on Mueller-Hinton agar supplemented with 2% NaCl (MH-NaCl agar) or on paper disc method agar supplemented with 5% defibrinated blood (PDM-blood agar). The Etest, multipoint inoculation test and disc diffusion test showed comparable sensitivity (0.96, 0.96 and 0.95, respectively) using an oxacillin breakpoint of > or = 0.5 mg/L or < or = 17 mm for the disc test, after incubation at 35 degrees C for 24 h on MH-NaCl agar. The sensitivity decreased for breakpoints > or = 1 mg/L and when PDM-blood agar was used instead of MH-NaCl agar.

Bacterial Proteins↗

[Studies of bovine Mycoplasma mastitis. 2. Testing of various culture media and culture methods for the isolation of Mycoplasma from milk samples].

The recipes Medium-I broth, Medium-B broth, and Weissenseemycoplasma broth as well as Medium-I agar, Medium-B agar, and MRL agar were tested for their applicability to culturing mycoplasma from milk samples, using the direct and indirect techniques. No dependable information on the occurrence of mycoplasma was obtainable from tested material unless several nutritive media and techniques were combined. Medium I, for which almost no imported substances were needed, was in no way inferior to common international nutritive substrates. Its use in conjunction with the indirect culturing technique is recommended for routine diagnosis.

Animals↗

Comparison of BACTEC PLUS blood culture media to BacT/Alert FA blood culture media for detection of bacterial pathogens in samples containing therapeutic levels of antibiotics.

Blood culture bottles with antimicrobial removal systems are recommended for patients who develop fever while on antibiotics. This study compared the ability of Becton Dickinson (Sparks, MD) BACTEC PLUS bottles and bioMerieux (Durham, NC) BacT/Alert FA bottles to effectively remove vancomycin, cefoxitin, ceftriaxone, cefepime, piperacillin-tazobactam, ampicillin, oxacillin, gentamicin, and a combination of gentamicin/penicillin, thus allowing bacterial pathogens to grow. Each bottle was spiked with 10 ml of human blood, antibiotic, and strains of organisms susceptible to the antibiotic evaluated. The organisms used were type strains and clinical isolates of Staphylococcus aureus (methicillin susceptible and resistant), Streptococcus pneumoniae, a viridans streptococcus, Enterococcus faecalis, Enterococcus faecium, Streptococcus agalactiae, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Testing was completed in triplicate, using 10 to 100 CFU/ml of organisms with various concentrations of each antibiotic. Two rounds of testing were completed per antibiotic/organism combination. Bottles were mixed and loaded onto their respective instruments as per the manufacturer's instructions. Antimicrobial removal was evaluated on the basis of time to detection of organism growth, for up to 5 days of incubation. Overall, the BacT/Alert FA system recovered 25.1% of strains from test bottles and 96.9% of strains from growth control bottles (no antibiotic added), and the BACTEC PLUS system recovered 95.1% of strains from test bottles and 100% of strains from growth control bottles. Both systems performed well in the detection of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa in the presence of gentamicin. In the presence of ceftriaxone, neither system was able to recover Streptococcus pneumoniae. The ability to remove vancomycin and cefoxitin was also determined by measuring antibiotic levels remaining in bottles after 1 h of incubation. The results demonstrated remaining levels of 72 to 90% of vancomycin and 71 to 72% of cefoxitin in the BacT/Alert system. For the BACTEC system, remaining levels were 0 to 30% of vancomycin and 0% of cefoxitin. Under these simulated conditions, the BACTEC PLUS system was superior to the BacT/Alert FA system in recovering gram-positive and gram-negative bacterial pathogens in the presence of beta-lactam antibiotics, gentamicin/penicillin, and vancomycin.

Anti-Bacterial Agents↗

Confrontation of an invasive (MO4) and a noninvasive (MDCK) cell line with embryonic chick heart fragments in serum-free culture media.

Confronting cultures of precultured embryonic chick heart fragments (PHF) with aggregates of malignant cells in vitro have been shown to be relevant for a number of aspects of tumor invasion in vivo. Preculture of the heart fragments, formation of cell aggregates, and subsequent culture of confronting pairs have so far been done only in serum-containing culture media. We describe here confronting cultures of PHF with invasive MO4 mouse cell aggregates or noninvasive MDCK dog kidney cell aggregates in serum-free media. Heart fragments precultured in the absence of serum seemed to be necrotic after confronting culture in serum-free media. However, preculturing in media supplemented with 10% fetal bovine serum allowed us to do subsequent confronting cultures in absence of serum. Cell aggregates were also prepared in serum-containing medium. MO4 cells occupied and replaced the heart tissue within 4 d, whereas MDCK cells remained at the periphery of the PHF. This indicates that serum-free confronting cultures can discriminate between invasive and noninvasive cells. The viability of individual PHF and cell aggregates cultured in the same way as in confrontations was ascertained by histology and by explantation and postculturing on a solid tissue culture substrate. Growth of the cultures was smaller in serum-free media than in media supplemented with 10% fetal bovine serum. The main advantage of serum-free culture conditions in vitro is the elimination of the influence of serum components on invasion, and the ability to examine the effect on invasion of drugs that are susceptible to inactivation by serum.

Animals↗

Evaluation of growth hormone production from GH1 cells in vitro: effect of culture media and time in culture.

Growth hormone production by a rat pituitary tumor cell line (GH1) was measured during lag, exponential, and plateau phases of growth in different culture media. Growth hormone secretion was low during lag and early exponential phase; it increased late in the exponential phase and continued to increase during the plateau phase. This biphasic pattern of growth hormone production was observed in all media and sera utilized. Both the doubling time and growth hormone production were influenced by the choice of media and sera. In addition, the length of time in culture affected the growth fraction with passage level 40 GH1 cells having a 79% growth fraction, whereas the growth fraction of passage level 100 cells was 95%. Using the population doubling time as a criterion for a choice of medium, F-10 medium supplemented with 20% fetal bovine serum consistently yielded the most rapid doubling time (32 hr), whereas Dulbecco's MEM supplemented with 15% horse serum and 2.5% fetal bovine serum yielded the greatest plateau cell density. Growth hormone secretion and the population doubling times were directly related to culture conditions including length of time in culture, choice of tissue culture media, choice of sera, and the phase of cell growth (lag, exponential or plateau).

Animals↗