Topoisomerase 2α status in invasive breast carcinoma - comparison of its clinical value according to immunohistochemical and fluorescence in situ hybridization methods of evaluation.

The main purpose of the study was to compare topoisomerase 2α (TOP2A) status in invasive breast carcinomas to the outcome of a therapy containing neoadjuvant treatment with anthracyclines (a combination chemotherapy treatment for breast cancer, namely AC [cyclophosphamide, doxorubicin]). To achieve these goals we created a method of evaluation with criteria based on two methods used in the present study (immunohistochemical [IHC] and fluorescence in situ hybridization [FISH]). The threshold for positive immunohistochemically evaluated status was set for all cases with: nuclear stain intensity score 3+ in 10% or more nuclei and nuclear stain intensity score 2+ in 50% or more nuclei. Our results suggest that TOP2A status may be used as a predictive factor for patient selection for protocols which include anthracyclines as one of the chemotherapeutics. Both methods, IHC and FISH, are suitable for implementation for diagnostic purposes, but IHC positive status measured according to the criteria presented above is the best predictor of longer disease-free survival (DFS) according to our study. Immunohistochemical also gave satisfactory results in all analyzed cases in comparison to only 60% of cases analyzed by FISH.

Muscle mitochondrial density after exhaustive exercise in dogs: prolonged restricted activity and retraining.

The effect of exhaustive treadmill exercise on mitochondrial density (MD) and ultrastructural changes in quadriceps femoris muscle was studied in 7 normal, healthy, male mongrel dogs (2-4 years, 15-20 kg) before and after restricted activity (RA) (2 months for 5 dogs, 5 months for 2 dogs), and following a subsequent 2-month exercise retraining period (2-months group). Mean (+/- S.E.) time to exhaustion in the 2-month group decreased from 177 +/- 11 min before to 90 +/- 16 min (delta = -49%, p < 0.05) after RA; retraining increased tolerance to 219 +/- 36 min (delta = +24%, p < 0.05) above the pre-RA and 143% (p < 0.05) above the post-RA time. Post-RA exhaustion time in the 5-months group was 25 and 45 min (35 +/- 10 min). Muscle samples taken after RA showed abnormalities indicative of degeneration, which were reversed by retraining. Resting MD decreased (p < 0.05) from a control level of 27.8% to 14.7% (2 months) and 16.3% (5 months), and was restored to 27.1% (NS) after retraining. Exhaustive exercise caused an increase in MD under control conditions and after RA, but not following retraining. Disruption of mitochondria after exercise was evident after 5-months confinement. Factors causing mitochondrial changes and eventually their disruption during exercise after restricted activity are not related as much to the state of fatigue as to the pre-exercise quality of the muscle modified by disease or training.

Craviten effect on the metabolism of stored platelets.

Craviten effect on platelets suspended in their own plasma was investigated by two methods of incubation with the drug: the fresh platelets were incubated for 18 h at 4 degrees C with Craviten added to plasma and platelets stored for 18 h at 4 degrees C without Craviten were incubated with Craviten at 37 degrees C for 1 h. For evaluation of the metabolic activity and function of the platelets, the levels of high-energy compounds ATP and ADP, the activity of hexokinase and pyruvate kinase and the amount of adenyl nucleotides released by the platelets after thrombin addition were measured, and the spontaneous aggregation of platelets and c-AMP were determined. The experiments demonstrated that Craviten prevented the fall of ATP level of the stored platelets, raised the activity of hexokinase and pyruvate kinase in the platelets, increased the amount of nucleotides released by the platelets in the release reaction. Craviten inhibited also increased spontaneous aggregation of stored platelets and raised c-AMP level.

[Effect of cytostatic agents on the metabolism and ultrastructure of blood platelets in man]. / Wplyw leków cytostatycznych na metabolizm i ultrastrukture krwinek plytkowych czlowieka.

The effect of cytostatic drugs used in the treatment of haemopoietic system diseases: cerubidin, hydroxyurea. cytosine arabinoside and their combinations on the platelet count, platelet ultrastructure, metabolic activity and participation in the processes of haemostasis was investigated. The experiments were done using two concentrations of the drugs: clinical corresponding to the maximal single dose and a fivefold as high concentration, It was found that the investigated cytostatic agents and their cominations in the first concentration had no effect of the platelet count, level of free nucleotides, activity of glucose-6-phosphate dehydrogenase and lactic dehydrogenase, and on the reaction of adenine nucleotides release from the platelets induced with thrombin. Using the higher concentration of the drugs it was observed that cerubidin and the combination of cytostatic agents tested changed the shape and ultrastructure of the platelets.

Electronmicroscopic histochemical investigation of the surface of the cell membrane of the renal tubules with the use of ruthenium red and lantanum nitrate.

The renal tubules were investigated with the use of ruthenium red and lanthanum nitrate to titrate the carbohydrate groups which are localized on the differentiated surface of the cell membranes and intercellular spaces. Ruthenium red visualized the surface coat which is tightly bound to the outer lamellae of the cell membrane. Lanthanum nitrate used in this investigations is a valuable marker of the intercellular spaces. The applied markers have visualized in the kidney the canals which are originated from the foldings of membranes of the tubular cells which adhere to the basal membrane. The markers used in combination with the fixative glutaraldehyde-paraformaldehyde give an electrondense envelope of the cell membrane which is more distinct as compared with specimen treated with glutaraldehyde and markers only.

Horseradish peroxidase application to the protein transport investigations and alkaline phosphatase ultralocalization at the kidney barrier level.

Horseradish peroxidase was used to the precise investigation of low molecular protein transport across the kidney barrier. Simultaneously on the parallel material alkaline phosphatase activity was revealed, as this enzyme is connected with the transport processes through cell membranes. The applied reaction diaminebenzidine has shown infiltration spots for low molecular protein at the glomercular barrier and visualized tubular reabsorption of peroxidase in proximal convoluted tubule. Alkaline phosphatase activity has been observed on the surface of podocytes foot processes. On the surface of proximal convoluted tubules brush border and in canals formed with cells basal membranes investigations high activity of alkaline phosphatase was also observed.

[Storage of platete concentrates at +4 degrees C and +22 degrees C]. / Przechowywanie koncentratów krwinek plytkowych w temperaturze +4 degrees C i +22 degrees C.

The relationship between the conditions of platelet preservation and their ultrastructure, metabolic activity, and the ability to carry out physiological functions was studied. It was found that storage of platelet concentrates at +4 degrees C for 72 hours reduced the number of platelets, changed their shape decreased the ATP level and the ability of release of adenine nucleotides in response to thrombin. The platelet at +22 degrees C for 72 hours preserved the initial number, normal ultrastructure, intact metabolic activity and the ability of participation in heamostasis processes.

Changes in the cellular membrane surface coat of lymphocytes and thymocytes after incubation in vitro with cystein as revealed with electronmicroscopy.

Changes in the cellular membrane surface coat of lymphocytes and thymocytes after incubation with cystein in vitro were revealed with electronmicroscope, while performing the reaction with Ruthenium Red and Concanavaline A. Lymphocytes and thymocytes not incubated with cystein to which reaction with Ruthenium red and Cocanavaline A was applied have shown a well developed and preserved surface coat of the cellular membrane. Contrary to this finding when lymphocytes and thymocytes were incubated with cystein and thereafter treated with Ruthenium Red and Concanavaline A no reaction product on the surface of the cellular membrane was observed. The experimental results could indicate on the influence of cystein on the glycoside bonds.