OBJECTIVE: Enzymatic fingerprinting of key enzymes of glucose metabolism is a valuable analysis tool in cell physiological phenotyping of plant samples. Yet, a similar approach for mammalian cell line samples is missing. In this study, we applied semi-high throughput enzyme activity assays that were originally designed for plant samples and tested their feasibility in extracts of six frequently used mammalian cell lines (Caco2, HaCaT, C2C12, HEK293, HepG2 and INS-1E). RESULTS: Enzyme activities for aldolase, hexokinase, glucose-6-phosphate dehydrogenase, phosphoglucoisomerase, phosphoglucomutase, phosphofructokinase could be detected in samples of one or more mammalian cell lines. We characterized effects of sample dilution, assay temperature and repeated freeze-thaw cycles causing potential biases. After careful selection of experimental parameters, the presented semi-high throughput methods could be established as useful tool for physiological phenotyping of cultured mammalian cells.

• Klíčová slova
• 96 well format, Aldolase, Enzyme assays, Glucose-6-phosphate dehydrogenase, Hexokinase, INS-1E, Phosphofructokinase, Phosphoglucoisomerase, Phosphoglucomutase,
• MeSH
• aldolasa metabolismus   MeSH
• buněčné linie MeSH
• buňky Hep G2 MeSH
• Caco-2 buňky MeSH
• enzymatické testy metody   MeSH
• fenotyp MeSH
• fosfofruktokinasy metabolismus   MeSH
• fosfoglukomutasa metabolismus   MeSH
• fosfotransferasy s alkoholovou skupinou jako akceptorem metabolismus   MeSH
• glukosa-6-fosfátdehydrogenasa metabolismus   MeSH
• glukosa metabolismus   MeSH
• glykolýza * MeSH
• HEK293 buňky MeSH
• hexokinasa metabolismus   MeSH
• lidé MeSH
• metabolismus sacharidů * MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• pilotní projekty MeSH
• studie proveditelnosti MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldolasa MeSH
• fosfofruktokinasy MeSH
• fosfoglukomutasa MeSH
• fosfotransferasy s alkoholovou skupinou jako akceptorem MeSH
• glukosa-6-fosfátdehydrogenasa MeSH
• glukosa MeSH
• hexokinasa MeSH
• phosphoglucokinase MeSH Prohlížeč

OBJECTIVE: To develop response criteria for adult dermatomyositis (DM) and polymyositis (PM). METHODS: Expert surveys, logistic regression, and conjoint analysis were used to develop 287 definitions using core set measures. Myositis experts rated greater improvement among multiple pairwise scenarios in conjoint analysis surveys, where different levels of improvement in 2 core set measures were presented. The PAPRIKA (Potentially All Pairwise Rankings of All Possible Alternatives) method determined the relative weights of core set measures and conjoint analysis definitions. The performance characteristics of the definitions were evaluated on patient profiles using expert consensus (gold standard) and were validated using data from a clinical trial. The nominal group technique was used to reach consensus. RESULTS: Consensus was reached for a conjoint analysis-based continuous model using absolute percent change in core set measures (physician, patient, and extramuscular global activity, muscle strength, Health Assessment Questionnaire, and muscle enzyme levels). A total improvement score (range 0-100), determined by summing scores for each core set measure, was based on improvement in and relative weight of each core set measure. Thresholds for minimal, moderate, and major improvement were ≥20, ≥40, and ≥60 points in the total improvement score. The same criteria were chosen for juvenile DM, with different improvement thresholds. Sensitivity and specificity in DM/PM patient cohorts were 85% and 92%, 90% and 96%, and 92% and 98% for minimal, moderate, and major improvement, respectively. Definitions were validated in the clinical trial analysis for differentiating the physician rating of improvement (P < 0.001). CONCLUSION: The response criteria for adult DM/PM consisted of the conjoint analysis model based on absolute percent change in 6 core set measures, with thresholds for minimal, moderate, and major improvement.

• MeSH
• alanintransaminasa metabolismus   MeSH
• aldolasa metabolismus   MeSH
• antirevmatika terapeutické užití   MeSH
• aspartátaminotransferasy metabolismus   MeSH
• dermatomyozitida farmakoterapie   metabolismus   patofyziologie   MeSH
• hodnocení výsledků péče pacientem MeSH
• hodnocení výsledků zdravotní péče MeSH
• kreatinkinasa metabolismus   MeSH
• L-laktátdehydrogenasa metabolismus   MeSH
• lidé MeSH
• logistické modely MeSH
• polymyozitida farmakoterapie   metabolismus   patofyziologie   MeSH
• průzkumy a dotazníky MeSH
• revmatologie MeSH
• společnosti lékařské MeSH
• svalová síla MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• konsensus - konference MeSH
• Geografické názvy
• Evropa MeSH
• Spojené státy americké MeSH
• Názvy látek
• alanintransaminasa MeSH
• aldolasa MeSH
• antirevmatika MeSH
• aspartátaminotransferasy MeSH
• kreatinkinasa MeSH
• L-laktátdehydrogenasa MeSH

OBJECTIVE: To develop response criteria for juvenile dermatomyositis (DM). METHODS: We analyzed the performance of 312 definitions that used core set measures from either the International Myositis Assessment and Clinical Studies Group (IMACS) or the Paediatric Rheumatology International Trials Organisation (PRINTO) and were derived from natural history data and a conjoint analysis survey. They were further validated using data from the PRINTO trial of prednisone alone compared to prednisone with methotrexate or cyclosporine and the Rituximab in Myositis (RIM) trial. At a consensus conference, experts considered 14 top candidate criteria based on their performance characteristics and clinical face validity, using nominal group technique. RESULTS: Consensus was reached for a conjoint analysis-based continuous model with a total improvement score of 0-100, using absolute percent change in core set measures of minimal (≥30), moderate (≥45), and major (≥70) improvement. The same criteria were chosen for adult DM/polymyositis, with differing thresholds for improvement. The sensitivity and specificity were 89% and 91-98% for minimal improvement, 92-94% and 94-99% for moderate improvement, and 91-98% and 85-86% for major improvement, respectively, in juvenile DM patient cohorts using the IMACS and PRINTO core set measures. These criteria were validated in the PRINTO trial for differentiating between treatment arms for minimal and moderate improvement (P = 0.009-0.057) and in the RIM trial for significantly differentiating the physician's rating for improvement (P < 0.006). CONCLUSION: The response criteria for juvenile DM consisted of a conjoint analysis-based model using a continuous improvement score based on absolute percent change in core set measures, with thresholds for minimal, moderate, and major improvement.

• MeSH
• alanintransaminasa metabolismus   MeSH
• aldolasa metabolismus   MeSH
• antirevmatika terapeutické užití   MeSH
• aspartátaminotransferasy metabolismus   MeSH
• cyklosporin terapeutické užití   MeSH
• dermatomyozitida farmakoterapie   metabolismus   patofyziologie   MeSH
• dítě MeSH
• glukokortikoidy terapeutické užití   MeSH
• hodnocení výsledků péče pacientem MeSH
• hodnocení výsledků zdravotní péče MeSH
• kreatinkinasa metabolismus   MeSH
• L-laktátdehydrogenasa metabolismus   MeSH
• lidé MeSH
• logistické modely MeSH
• methotrexát terapeutické užití   MeSH
• mladiství MeSH
• prednison terapeutické užití   MeSH
• průzkumy a dotazníky MeSH
• reprodukovatelnost výsledků MeSH
• revmatologie MeSH
• rituximab terapeutické užití   MeSH
• společnosti lékařské MeSH
• svalová síla MeSH
• výsledek terapie MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• Publikační typ
• časopisecké články MeSH
• konsensus - konference MeSH
• Geografické názvy
• Evropa MeSH
• Spojené státy americké MeSH
• Názvy látek
• alanintransaminasa MeSH
• aldolasa MeSH
• antirevmatika MeSH
• aspartátaminotransferasy MeSH
• cyklosporin MeSH
• glukokortikoidy MeSH
• kreatinkinasa MeSH
• L-laktátdehydrogenasa MeSH
• methotrexát MeSH
• prednison MeSH
• rituximab MeSH

Sturgeon and paddlefish (Acipenseriformes), the source of roe consumed as caviar, are a unique and commercially valuable group of ancient fishes. In this study, comparative proteomics was used to analyze protein profiles of spermatozoa from five sturgeon species and one paddlefish: Siberian sturgeon (Acipenser baerii), sterlet (A. ruthenus), Russian sturgeon (A. gueldenstaedtii), starry sturgeon (A. stellatus), beluga (Huso huso), and Mississippi paddlefish (Polyodon spathula). Protein profiles of spermatozoa were determined by isoelectric focusing and two-dimensional electrophoresis (2-DE) high-resolution gels. The peptides, previously selected by 2-DE analysis as potentially species-specific, were obtained by "in-gel" tryptic digestion, followed by matrix-associated laser desorption/ionization time-of-flight/mass spectrometry (MALDI-TOF/MS). Among the 23 protein spots selected, 14 were identified as isoforms of enolase B present in all species, but with different isoelectric points or molecular mass. Exceptions were A. ruthenus and H. huso, species with a close phylogenetic relationship. Glycerol-3-phosphate dehydrogenase was detected exclusively in P. spathula. Phosphoglycerate kinase was detected only in A. ruthenus and H. huso, and 3 additional proteins (fructose bisphosphate aldolase A-2, glycogen phosphorylase type IV and glyceraldehyde-3-phosphate dehydrogenase) were found exclusively in A. gueldenstaedtii and H. huso. This study points to the application of proteomics for differential characterization and comparative studies of acipenseriform species at the molecular level.

• MeSH
• 2D gelová elektroforéza metody   MeSH
• aldolasa genetika   metabolismus   MeSH
• biologické markery metabolismus   MeSH
• druhová specificita MeSH
• fosfoglycerátkinasa genetika   metabolismus   MeSH
• fosfopyruváthydratasa genetika   metabolismus   MeSH
• glycerolfosfátdehydrogenasa genetika   metabolismus   MeSH
• isoelektrická fokusace metody   MeSH
• izoenzymy genetika   metabolismus   MeSH
• peptidové mapování metody   MeSH
• peptidy genetika   metabolismus   MeSH
• proteomika metody   MeSH
• ryby klasifikace   genetika   metabolismus   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• spermie metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Geografické názvy
• Mississippi MeSH
• Sibiř MeSH
• Názvy látek
• aldolasa MeSH
• biologické markery MeSH
• fosfoglycerátkinasa MeSH
• fosfopyruváthydratasa MeSH
• glycerolfosfátdehydrogenasa MeSH
• izoenzymy MeSH
• peptidy MeSH

We have shown previously that iron deprivation significantly stimulates the uptake of non-transferrin ferric iron from ferric citrate by erythroleukemia K562 cells and that this stimulation depends on protein synthesis. However, we have not detected increased expression of any known iron transport protein (Kovar J. et al. (2006) Blood Cells Mol Dis 37:95-99). Therefore, in order to identify membrane proteins of K562 cells with increased expression under iron deprivation, we employed the isolation of membrane proteins by two-phase partitioning system, protein separation by high-resolution 2D electrophoresis, computer differential analysis, and tandem mass spectrometry. Employing these techniques we identified two proteins with statistically significant upregulation, i.e., aldolase A (ALDA) and voltage-dependent anion channel 2 (VDAC2). The upregulation of aldolase A and VDAC2 in K562 cells under iron deprivation was also confirmed by western blot analysis. This is the first time when the control of aldolase A and VDAC2 levels by iron status of the cell is demonstrated.

• MeSH
• aldolasa genetika   metabolismus   MeSH
• buňky K562 * MeSH
• deficit železa * MeSH
• lidé MeSH
• napětím ovládaný aniontový kanál 2 genetika   metabolismus   MeSH
• regulace genové exprese * MeSH
• upregulace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aldolasa MeSH
• napětím ovládaný aniontový kanál 2 MeSH
• VDAC2 protein, human MeSH Prohlížeč

AIMS: Lachnospira multiparus belongs to the main rumen pectinolytic bacteria. Its carbohydrate metabolism was studied in growth experiments on laboratory fermenters, and using assays of activities of enzymes involved in pectin fermentation. METHODS AND RESULTS: The type strain of this species and two substrates were used. Lachnospira multiparus ATCC 19207 grew on pectin and glucose at a similar rate and had no preference for one or the other substrate. Pectin-grown cultures, however, produced significantly more acetate and less formate, lactate, ethanol, hydrogen, cell dry matter and protein than corresponding cultures grown on glucose. Extracellular exopectate lyase (EC 4.2.2.9) was the principal enzyme degrading the pectin macromolecule. Cell extracts possessed 2-keto-3- deoxy-6-phosphogluconate aldolase (EC 4.1.2.14) and fructosediphosphate aldolase (EC 4.1.2.13) activity. The former enzyme catalyses the final reaction in the Entner-Doudoroff pathway; the latter is the key enzyme of glycolysis and the pentose phosphate pathway. CONCLUSION: These results are consistent with the assumption that acidic products of pectin degradation are catabolized via a modified Entner-Doudoroff pathway. Phosphogluconate was not metabolized by cell extracts of the strain studied. SIGNIFICANCE AND IMPACT OF THE STUDY: This suggests that the conventional Entner-Doudoroff pathway of glucose utilization does not operate in this bacterium, presumably because of the lack of 6-phosphogluconate dehydrase (EC 4.2.1.12) activity.

• MeSH
• aldolasa metabolismus   MeSH
• bachor mikrobiologie   MeSH
• fermentace MeSH
• glukonáty metabolismus   MeSH
• glukosa metabolismus   MeSH
• gramnegativní anaerobní bakterie enzymologie   růst a vývoj   metabolismus   MeSH
• pektiny metabolismus   MeSH
• pentózofosfátový cyklus MeSH
• polysacharid-lyasy izolace a purifikace   metabolismus   MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 2-keto-3-deoxy-6-phosphogluconate MeSH Prohlížeč
• aldolasa MeSH
• glukonáty MeSH
• glukosa MeSH
• pectate disaccharide-lyase MeSH Prohlížeč
• pektiny MeSH
• polysacharid-lyasy MeSH

The effect of the system of mating and the way of killing on the aldolase activity in the tissue of rabbits was the task of this research work. The research work was realized in 201 New Zealand white rabbits aged 140 days, mated outbred and inbred. Two methods of killing were used: classical method, which was based on the stroke and break the spinal cord; and killing with the help of treating with an i/m infection of 10 per cent of magnesium sulphate (2 ml per 1 kg of weight). In blood serum and in liver homogenates, kidneys and dorsal muscle the aldolase activity was determined by the method of Bruns. No significant differences in the aldolase activity between outbred and inbred rabbits were found. Statistically high significant differences, conditioned by killing method, were stated in activity of enzyme.

• MeSH
• aldolasa metabolismus   MeSH
• králíci metabolismus   MeSH
• síran hořečnatý farmakologie   MeSH
• zvířata MeSH
• Check Tag
• králíci metabolismus   MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• aldolasa MeSH
• síran hořečnatý MeSH

The authors performed a comparative biochemical study of some enzymes of lysosomic origin (hyaluronidase, N-acetyl-beta-D-glucosaminidase, beta-glucosidase, beta-galatosidase and acid phosphatase), of the state of enzyme substrate system N-acetylneuraminic acid---aldolase of neuramic acid and of the activity of lactatedehydrogenase (soluble in cytosol and bound on mitochodria) in the liver, lungs and blood serum of rats at various regimens of the inhalation action of CCl4. On the basis of results obtained they determined the biological importance of the change of activity of enzymes differently localized in cells at the adaptation of an organisme to the noxious action of CCl4.

• MeSH
• aldolasa metabolismus   MeSH
• chlorid uhličitý farmakologie   MeSH
• cytoplazma enzymologie   MeSH
• hydrolasy metabolismus   MeSH
• játra enzymologie   ultrastruktura   MeSH
• krysa rodu Rattus MeSH
• L-laktátdehydrogenasa metabolismus   MeSH
• lyzozomy enzymologie   MeSH
• plíce enzymologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldolasa MeSH
• chlorid uhličitý MeSH
• hydrolasy MeSH
• L-laktátdehydrogenasa MeSH

Biotin deficiency resulted in an increased growth rate of Aspergillus nidulans. The activities of hexokinase and aldolase were not much changed during the growth cycle, but activities of glucose-6-phosphate dehydrogenase and NADP-linked glutamate dehydrogenase increased significantly during the exponential phase. This change was remarkable during biotin deficiency. In contrast to the higher growth rate and respiration rate during biotin deficiency the activities of NAD(P)H oxidoreductases were low. An inverse relationship between the activity of tyrosinase and melanin content was observed. A role of the DOPA-DOPA-quinone system in maintaining culture growth is suggested.

• MeSH
• aldolasa metabolismus   MeSH
• Aspergillus nidulans enzymologie   růst a vývoj   metabolismus   MeSH
• biotin biosyntéza   MeSH
• dihydroxyfenylalanin metabolismus   MeSH
• glukosa-6-fosfátdehydrogenasa metabolismus   MeSH
• glukosa metabolismus   MeSH
• glutamátdehydrogenasa metabolismus   MeSH
• hexokinasa metabolismus   MeSH
• melaniny biosyntéza   MeSH
• mutace MeSH
• NADH, NADPH oxidoreduktasy metabolismus   MeSH
• spotřeba kyslíku MeSH
• tyrosinasa metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldolasa MeSH
• biotin MeSH
• dihydroxyfenylalanin MeSH
• glukosa-6-fosfátdehydrogenasa MeSH
• glukosa MeSH
• glutamátdehydrogenasa MeSH
• hexokinasa MeSH
• melaniny MeSH
• NADH, NADPH oxidoreduktasy MeSH
• tyrosinasa MeSH

• MeSH
• aldolasa metabolismus   MeSH
• biologický transport MeSH
• fruktosa biosyntéza   metabolismus   MeSH
• hexokinasa metabolismus   MeSH
• intestinální absorpce MeSH
• inzulin metabolismus   MeSH
• játra enzymologie   metabolismus   MeSH
• ledviny enzymologie   MeSH
• lidé MeSH
• pankreas metabolismus   MeSH
• psi MeSH
• sacharosa metabolismus   MeSH
• sekrece inzulinu MeSH
• svaly enzymologie   MeSH
• trávení MeSH
• tuková tkáň enzymologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• aldolasa MeSH
• fruktosa MeSH
• hexokinasa MeSH
• inzulin MeSH
• sacharosa MeSH