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Culture Procedure (culture + procedure)

Distribution by Scientific Domains

Medical Sciences	75%

Selected Abstracts

Effects of rapamycin on accumulation of , -, , - and , -globin mRNAs in erythroid precursor cells from , -thalassaemia patients

EUROPEAN JOURNAL OF HAEMATOLOGY, Issue 5 2006
Eitan Fibach
Abstract:, We studied the effects of rapamycin on cultures of erythroid progenitors derived from the peripheral blood of 10 , -thalassaemia patients differing widely with respect to their potential to produce foetal haemoglobin (HbF). For this, we employed the two-phase liquid culture procedure for growing erythroid progenitors, high performance liquid chromatography for analysis of HbF production and reverse transcription polymerase chain reaction for quantification of the accumulation of globin mRNAs. The results demonstrated that rapamycin induced an increase of HbF in cultures from all the , -thalassaemia patients studied and an increase of their overall Hb content/cell. The inducing effect of rapamycin was restricted to , -globin mRNA accumulation, being only minor for , -globin and none for , -globin mRNAs. The ability of rapamycin to preferentially increase , -globin mRNA content and production of HbF in erythroid precursor cells from , -thalassaemia patients is of great importance as this agent (also known as sirolimus or rapamune) is already in clinical use as an anti-rejection agent following kidney transplantation. These data suggest that rapamycin warrants further evaluation as a potential therapeutic drug in , -thalassaemia and sickle cell anaemia. [source]

Microbiologic Diagnostics at Titanium Implants

CLINICAL IMPLANT DENTISTRY AND RELATED RESEARCH, Issue 4 2003
Åsa Leonhardt DDS
ABSTRACT Background: The microbiota found at periimplant lesions have been shown to contain putative periodontal pathogens as well as opportunistic species such as Staphylococcus spp, enterics, and Candida spp. Therefore, a microbiologic diagnosis may be of value as guidance before treatment of such lesions. Purpose: The aim of this study was to evaluate the prevalence of some putative pathogens associated with long-term fol-lowed-up cases using two different microbiologic procedures. Malerials and Methods: Fifteen subjects contributed with plaque samples from teeth and implants; these were analyzed with respect to 18 putative periimplant pathogens using cultural methods and a deoxyribonucleic acid DNA-DNA hybridization technique. Results: The number of individuals positive for the analyzed pathogens was similar in samples taken from teeth and implants when analyzed with the DNA-DNA hybridization technique. When comparing detection frequency by culture procedure and by "checkerboard" technique at implants, the number of individuals positive for these species was lower with the traditional culture technique than with the checkerboard analyses. Using a higher cutoff point (4) with the checkerboard technique, the number of positive individuals was generally lower than that found with the culture technique. When comparing the techniques on an implant site level, the prevalence obtained by culture was lower for all analyzed species. If the specific species were present in the samples analyzed by the checkerboard technique, they were present only in every second sample analyzed with the culture technique. The high specificity values showed that if the checkerboard technique did not detect any Porphyromonas gingivalis, Prevotla intermedia, Actinobadllus actinomycetem-comitans, or Fusobacterium nudeatum, the bacteria were also undetectable by the culture technique. The two methods therefore did not overlap but did supplement each other. Conclusions: Based on the current results it is recommended that the technique used when analyzing microbiota around titanium implants should be a combination of the two protocols mentioned as they seem to give the most comprehensive outcome when used together. [source]

Improved detection of chromosomal abnormalities in chronic lymphocytic leukemia by conventional cytogenetics using CpG oligonucleotide and interleukin-2 stimulation: A Belgian multicentric study,

GENES, CHROMOSOMES AND CANCER, Issue 10 2009
Natalie Put
We performed a multicentric study to assess the impact of two different culture procedures on the detection of chromosomal abnormalities in 217 consecutive unselected cases with chronic lymphocytic leukemia (CLL) referred for routine analysis either at the time of diagnosis (n = 172) or during disease evolution (n = 45). Parallel cultures of peripheral blood or bone marrow were set up with the addition of either the conventional B-cell mitogen 12- O -tetradecanoyl-phorbol-13-acetate (TPA) or a combination of CpG oligonucleotide (CpG) and interleukin-2 (IL-2). Cytogenetic analyses were performed on both cultures. Clonal abnormalities were identified in 116 cases (53%). In 78 cases (36%), the aberrant clone was detected in both cultures. Among these, the percentages of aberrant metaphases were similar in both conditions in 17 cases, higher in the CpG/IL-2 culture in 43 cases, and higher in the TPA culture in 18 cases. Clonal aberrations were detected in only one culture, either in CpG/IL-2 or TPA in 33 (15%) and 5 (2%) cases, respectively. Taken together, abnormal karyotypes were observed in 51% with CpG/IL-2 and 38% with TPA (P < 0.0001). Application of FISH (n = 201) allowed the detection of abnormalities not visible by conventional cytogenetic analysis in 80 cases: del(13q) (n = 71), del(11q) (n = 5), +12 (n = 2), del(14q) (n = 1), and del(17p) (n = 1). In conclusion, our results confirm that CpG/IL-2 stimulation increases the detection rate of chromosomal abnormalities in CLL compared with TPA and that further improvement can be obtained by FISH. However, neither conventional cytogenetics nor FISH detected all aberrations, demonstrating the complementary nature of these techniques. © 2009 Wiley-Liss, Inc. [source]

Optimization of the expression of recombinant human activin A in the yeast Pichia pastoris

BIOTECHNOLOGY PROGRESS, Issue 2 2010
Dale Fredericks
Abstract We report a new procedure to express recombinant human activin A using the methanolic yeast, Pichia pastoris. Optimization of culture procedures has involved comprehensive examination of the effects of culture vessel shape, volume of broth in the induction and expression cultures, methanol concentration, culturing temperature, and pH of the expression cultures. After this optimization, as well as modification of the native cleavage sites, a laboratory scale procedure has been established which routinely produced 2,10 mg/L amounts of this vital growth factor in the highly efficient, eukaryotic yeast system. This system avoids the need to produce this protein and similar TGF-, proteins in mammalian cell lines which, in addition to being costly, produce many native binding partners of these cystine knot proteins, a factor which can dramatically affect yields of the target protein. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source]