12 research outputs found
Carbohydrate Composition of Endotoxins from R-type Isogenic Mutants of Shigella sonnei Studied by Capillary Electrophoresis and GC-MS
S3 Code. Matlab code to simulate epidemic when hosts perform a Levy wal
Carbohydrate Composition of Endotoxins from R-type Isogenic Mutants of Shigella sonnei Studied by Capillary Electrophoresis and GC-MS
The carbohydrate composition of the rough-type endotoxins (lipopolysaccharides, LPSs) from
Shigella sonnei mutant strains (Shigella sonnei phase II - 4303, 562H, R41 and 4350) was investigated by
capillary electrophoresis and GC-MS. The monosaccharides obtained by hydrolysis were determined by
capillary electrophoresis combined with laser induced fluorescence detection (CE-LIF) after labeling with
8-aminopyrene-1,3,6-trisulfonic acid (APTS) and by GC-MS as alditol-acetate derivatives. It was obtained
that the lipopolysaccharides of the isogenic rough mutants are formed in a step-like manner, containing
no heptose (4350), one D-glycero-D-mannoheptose (562H), or two or three L-glycero-Dmannoheptoses
(R41, 4303, respectively) in the deep core region. Besides the heptoses, the longest
LPS from the mutant Shigella sonnei 4303 contains hexoses, such as glucoses and galactoses, in a proportion
of approximately 3:2. This study provides a comprehensive comparison of the variability in
the carbohydrate part of the rough endotoxins extracted from Shigella sonnei isogenic mutants.
(doi: 10.5562/cca1795
The impact of the crisis and recovery on HR and knowledge management in focus - a Hungarian-Slovakian comparison 2009
The world slowly begins to recover from the economic crisis. According to the forecast of the most recent Davos World Economic Forum (2011), the entire world economy can anticipate a 6.5%, while the developed world - including the transition countries - an approximately 2.5-3%GDP growth. It was not easy to get to this point. Since October 2008, the actors of the world economy got many
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slaps in the face
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(ILO, 2009). Since October 2008, the global economy are very much
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clout
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for it lacks the good character and prudence (Palinkas, 2009). The crisis itself particularly affected to a large extent economies of Eastern Europe, choosen a neo-liberal way. (Csaba, 2009; Kiss, 2009 and Szelényi, 2009). Up to this day, we have a relatively small amount of specific data and few surveys (Bóday, 2009) on how the different companies and institutions survived the crisis from a HR and knowledge management point of view, what methods they used to mitigate the effects of the crisis. In the present study, we give a summary of our relevant observations gained from an empirical survey we conducted between October and December 2009 among Hungarian and Slovakian companies
Green Fluorescent Protein-Based Viability Assay in a Multiparametric Configuration
Green fluorescent protein (GFP) is considered to be suitable for cell viability testing. In our study, GFP transfected A549 lung carcinoma cell line was treated with sodium fluoride (NaF), cycloheximide (CHX) and ochratoxin A (OTA). GFP fluorescence, intracellular ATP, nucleic acid and protein contents were quantified by a luminescence microplate assay developed in our laboratory. Flow cytometry was used to confirm the findings and to assess the intensity of GFP during different types of cell death. A 24 h NaF and CHX exposure caused a dramatic decrease in ATP contents (p < 0.05) compared with those of the controls. GFP fluorescence of the cells was in close correlation with total protein; however, GFP/ATP increased at NaF and decreased at CHX treatments (p < 0.05). ATP/protein and ATP/propidium iodide (PI) were largely decreased at NaF exposure in a dose-dependent manner (p < 0.05), while CHX and OTA showed markedly fewer effects. Both treatments caused apoptosis/necrosis at different rates. NaF induced mainly late apoptosis while OTA, mainly apoptosis. CHX effects varied by the incubation time with 100-fold elevation in late apoptotic cells at 24 h treatment. GFP intensity did not show a significant difference between live and apoptotic populations. Our results suggest when using GFP, a multiparametric assay is necessary for more precise interpretation of cell viability
A One-Step Extraction and Luminescence Assay for Quantifying Glucose and ATP Levels in Cultured HepG2 Cells
A fluorescence-based enzymatic microplate intracellular glucose assay was designed and fully validated. The method was tested in a hepatocellular cancer cell line (HepG2). Our novel one-step extraction reagent gave stable cell lysates for glucose, adenosine triphosphate (ATP), and total protein determination from the same sample. Limit of detection for glucose was 0.13 µM (26 pmol/well), which is superior to commercially available glucose assays. Both intra- and interday assay imprecision in HepG2 cultures were less than 12% coefficient of variance (CV). In cell lysates spiked with glucose, recovery at two levels varied between 83.70% and 91.81%, and both linearity and stability were acceptable. HepG2 cells treated with agents affecting glucose uptake/metabolism (phloretin, quercetin, quercetin-3′-sulfate, NaF, 3-bromopyruvate, NaN3, oligomycin A, ochratoxin A, cytochalasin B, and anti-GLUT1 antibody) showed dose-dependent changes in glucose and ATP levels without total protein (cell) loss. Finally, we performed flow cytometric glucose uptake measurement in the treated cells using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose fluorescent glucose analog. Glucose uptake did not always mirror the intracellular glucose levels, which most likely reflects the differences between the two methodologies. However, interpreting data obtained by both methods and taking ATP/protein levels at the same time, one can get information on the mode of action of the compounds
Diagnostic relevance of urinary steroid profiles on ovarian granulosa cell tumors: two case reports
Abstract Background Granulosa cell tumor of the ovary is the most frequent sex cord stromal tumor and represents 2 to 5% of all primary ovarian cancers. Ovarian granulosa cell tumor is a malignant tumor with slow progression and in some cases this tumor is hormonally active. The recurrence of granulosa cell tumor often happens after 5 years. Case presentation We describe two cases of postmenopausal women with adult-type granulosa cell tumors of the ovary. Patient 1 is a 49-year-old European woman with a recurrent tumor; patient 2 is a 55-year-old European woman without recurrence of tumor. Urinary steroid profiles of patient 1 were monitored during a 5-year period starting from before an operation (13 samples). In patient 2, the urinary steroid profiles were monitored during a 3-year period starting from after an operation (six samples). The 24-hour urinary samples were examined and the urinary concentration of 20 androgen, progesterone, and corticoid metabolites was quantitatively determined by gas chromatography-mass spectrometry with selected ion-monitoring mode. Conclusions Based on these cases a correlation could be observed between increased levels of the urinary steroids and the recurrence of ovarian granulosa cell tumor; therefore, we concluded that a urinary steroid profile could be a more effective method to follow-up such patients compared to the traditional serum hormones determinations supplemented with conventional tumor markers