Immunohistochemical Analysis of ATRX, IDH1 and p53 in Glioblastoma and Their Correlations with Patient Survival

Glioblastoma (GBM) can be classified into molecular subgroups, on the basis of biomarker expression. Here, we classified our cohort of 163 adult GBMs into molecular subgroups according to the expression of proteins encoded by genes of alpha thalassemia/mental retardation syndrome X-linked (ATRX), isocitrate dehydrogenase (IDH) and TP53. We focused on the survival rate of molecular subgroups, depending on each and various combination of these biomarkers. ATRX, IDH1 and p53 protein expression were evaluated immunohistochemically and Kaplan-Meier analysis were carried out in each group. A total of 15.3% of enrolled GBMs demonstrated loss of ATRX expression (ATRX-), 10.4% expressed an aberrant IDH1 R132H protein (IDH1+), and 48.4% exhibited p53 overexpression (p53+). Survival differences were statistically significant when single protein expression or different combinations of expression of these proteins were analyzed. In conclusion, in the case of single protein expression, the patients with each IDH1+, or ATRX-, or p53- GBMs showed better survival than patients with counterparts protein expressed GBMs. In the case of double protein pairs, the patients with ATRX-/p53-, ATRX-/IDH1+, and IDH1+/p53- GBMs revealed better survival than the patients with GBMs with the remained pairs. In the case of triple protein combinations, the patients with ATRX-/p53-/IDH+ showed statistically significant survival gain than the patients with remained combination of proteins-expression status. Therefore, these three biomarkers, individually and as a combination, can stratify GBMs into prognostically relevant subgroups and have strong prognostic values in adult GBMs.

Impact of copper on the oxidative metabolism of the fry of common carp, Cyprinus carpio (Linn.) at different pH.

In order to evaluate the impact of copper on the energetics of a fish, the levels of glucose, glycogen, pyruvate and lactate, the rate of tissue oxygen consumption and the activities of glycogen phosphorylase, isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) were estimated in the whole body of the fry of Cyprinus carpio immediately after 1, 7, 15 and 30 days on exposure to a sublethal concentration of copper 0.08 mgl(-1) at pH 7.5 (normal), 6.0 (weak acidic) and 9.0 (weak alkaline). Aprogressive increase in glucose level and glycogen phosphorylase activity with the corresponding decrease in glycogen level over the time of exposure at pH 7.5 indicated glycogenolysis. Increase in the rate of oxygen consumption, pyruvate level and ICDH and SDH activities at days 1 and 7 (day 1 > 7) followed by their decrease at days 15 and 30 (day 15 < 30) at pH 7.5 indicated an initial elevation in the energetics of the fish fry with a gradual suppression of it on prolonged exposure. During this period the animal might have relied more on energetically less efficient glycolysis as evident by the progressive increase in the level of lactate and LDH activity. The degree of glycogenolysis was relatively more at pH 6.5 than at pH 7.5. At that pH, a progressive decrease in glucose level with an increase in the pyruvate and lactate levels and in LDH activity and a decrease in the rate of oxygen consumption and ICDH and SDH activities revealed greater reliance of the fish on anaerobic glycolysis than on oxidative metabolism. At pH 9.0 also the fish fry initially exhibited glycogenolysis, but gradually it came to normal on day 30 (day 1 > 7 > 15 > 30). Decrease in the glucose level, increase in pyruvate level, rate of oxygen consumption, and ICDH and SDH activities at all the days of exposure suggested an elevation in oxidative metabolism, but it also came to normal on prolonged exposure. Even the lactate level and LDH activity initially increased but gradually reached to normal on day 30. These results indicated that copper suppresses the energetics of the fish fry at pH 6.0, elevates at pH 9.0 relative to the changes at pH 7.5 suggesting that the toxicity of copper is dependent on pH of the water.

Acridine orange staining and viability of the coccoid form of Campylobacter upsaliensis.

Conversion of Campylobacter upsaliensis to the coccoid form during aerobic incubation at 37 degrees C was not prevented by treatment with chloramphenicol and was accompanied by severe decreases in isocitrate dehydrogenase activity and oxygen uptake. Although the coccoid forms fluoresced orange-red by acridine orange staining, agarose gel electrophoresis indicated an extensive degradation of the ribosomal RNA. This suggests that acridine orange staining may not be a good indicator of viability and that the coccoid form of C. upsaliensis at 37 degrees C is degenerative rather than part of the life cycle.

Short-term effects of thyroid hormones on lipogenic enzymes and 14C-acetate incorporation into various lipid classes: in vivo and in vitro studies.

Short-term effect of 3,5,3'-triiodothyronine (T3) and 3,5-diiodothyronine (T2) on lipid metabolism in the liver of Anabas testudineus was examined. In vivo injections of both T3 and T2 at a concentration of 10 ng/g body weight increased malic enzyme (ME), glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) activity compared to 6-propylthiouracil (6-PTU) treated group. Treatment of 6-PTU results in the accumulation 14C-acetate into fat and thyroid hormones' treatment reduce it. In vitro experiments show that malic enzyme activity is augmented only by high concentration of T3 (10(-7) M) where as all concentrations of T2 increase its activity. In vitro studies with T3 showed a biphasic effect on cholesterol content. Conversely T2 in vitro, reduced cholesterol content with all concentrations. From these results it can be concluded that both T3 and T2 have short-term effect on lipid metabolism in Anabas.

Metabolic studies on mycobacteria. III. Demonstration of key enzymes of TCA cycle in M. leprae.

Cell free extracts of armadillo derived M. leprae, M. phlei, M. smegmatis and normal armadillo liver were analysed for the two key enzymes of TCA cycle. Aconitase activity was assayed in the presence of inhibitor fluorocitrate and it was observed that cell free extracts from cultivable mycobacteria as well as aramadillo derived M. leprae had this enzyme activity and 66-82% of this activity was inhibited by 0.1 mM fluorocitrate. 74% of M. leprae derived enzyme activity was inhibited by fluorocitrate in contrast with armadillo derived enzyme which was only 29% inhibited by fluorocitrate. PAGE separation of cell free extracts and staining for Isocitrate dehydrogenase (ICD) activity showed that an additional bond of ICD activity was demonstrable in the cell free extracts of armadillo derived M. leprae and this was NADP dependent. The mobility (ef) of this band of activity was in the same range as ICD from cultivable mycobacteria and much lower than ICD from normal armadillo liver. From this study and from the previously reported work, it is concluded that like other mycobacteria TCA cycle is operative in M. leprae.