Inhibitory effect of combined treatment with the aromatase inhibitor exemestane and tamoxifen on DMBA-induced mammary tumors in rats

The antitumor effect of exemestane (FCE 24304), an irreversible aromatase inhibitor, given alone or in combination with tamoxifen, was investigated in rats with 7, 12-dimethylbenzanthracene (DMBA)-induced mammary tumors. The compounds were given once daily, 6 days a week for 4 weeks. Exemestane, given at the dose of 20 mg/kg/day s.c., induced 26% complete (CR) and 18% partial (PR) tumor regressions, compared to 0% CR and 6% PR observed in controls. Tamoxifen, given at 1 mg/kg/day p.o., induced 16% CR and 13% PR. The combined treatment caused 41% CR and 16% PR, thus resulting in a higher antitumor effect than either single treatment. The apperance of new tumors was reduced by each single treatment and almost totally prevented by the combined treatment. Serum prolactin (PRL) levels, assayed 4 h after the last dose, were unchanged in the group treated with the combination, whereas tamoxifen alone caused a slight increase of serum PRL. These results indicate that estrogen deprivation through aromatase inhibition and estrogen receptor antagonism causes a better inhibition of DMBA-induced mammary tumors than either treatment modality alone.     

Assignment of the prealbumin (PALB) gene (familial amyloidotic polyneuropathy) to human chromosome region 18q11.2–q12.1

Summary The assignment of the human prealbumin (PALB) gene to chromosome region 18q11–q12.1 has been achieved using a human genomic probe in the study of human-mouse somatic cell hybrids and by in situ hybridization. Because familial amyloidotic polyneuropathy was reported previously to be due to a mutation in prealbumin, it can be inferred that the gene for this disorder also maps to 18q11.2–q12.1.     

Free radicals promote modifications in plasma high-density lipoprotein: nuclear magnetic resonance analysis.

The behavior of high-density lipoprotein (HDL) after free-radical-mediated oxidation was studied by incubating plasma HDL with chemical oxidizing systems (Cu++) in conditions similar to those used for low-density lipoprotein (LDL) chemical oxidation. Nuclear magnetic resonance (NMR) spectroscopy (1H and 31P) was used to evaluate the degree of oxidation and to characterize the oxidized products. The almost complete loss of polyunsaturated systems together with an appreciable decrease in choline peak demonstrates large-scale HDL-lipid degradation. The appearance of epoxide systems on fatty chains and the identification of oxidized cholesterol derivatives as cholesterol 5 alpha,6 alpha-epoxide, 5 beta,6 beta-epoxide, 7-keto, and 25-hydroxy confirm this picture. Phospholipid analysis indicates an alteration of the phospholipid profile in lyso-phosphatidylcholine (Lyso-PC) production and the disappearance of phosphatidylethanolamine (PE). This study shows that HDL is extensively degraded although there are no large variations in the classical oxidative monitors, lipid hydroperoxide (LPO) and thiobarbituric acid reactive substance (TBARS). Our results suggest that HDL is significantly modified when submitted to an oxidative process.     

Time-Lapse Cinemicrographic Studies of X-Irradiated HeLa S3 Cells: II. Cell Fusion

Analysis of time-lapse cinemicrographs of X-irradiated HeLa S3 cells has shown that the incidence of cell fusion was increased from 0.9% (following 1267 divisions) in control cells to an average of 22% (following 655 divisions) in cells irradiated with 500 rad doses of 220 kv X-rays. The incidence depended on the stage of the generation cycle at which the parent cells were irradiated. It was nearly constant in the first three postirradiation generations. Fusion occurred at all stages of the generation cycle, but preferentially during the first 20%. Cells undergoing fusion progressed more slowly through the generation cycle and had a higher probability of disintegrating than did irradiated cells that did not fuse. The occurrence of fusion was clonally distributed in the population. It took place only between sister (or closely related) cells. Protoplasmic bridges were often visible between sister cells prior to fusion. Giant cells arose only as a result of fusion. The incidence of multipolar divisions, though higher than in unirradiated cells, was only 5.5% in cultures irradiated with 500 rads. Fusion occurred following 85% of the multipolar divisions and was often followed by a multipolar division.     

Apolipoprotein A-I metabolism in subjects with a PstI restriction fragment length polymorphism of the apoA-I gene and familial hypoalphalipoproteinemia

Familial hypoalphalipoproteinemia (hypoalpha), characterized by a decreased high density lipoprotein level, is associated with an increased incidence of premature cardiovascular disease. Restriction fragment length polymorphism analysis of genomic DNA has detected a polymorphism for the PstI restriction endonuclease near the apoA-I gene, with either a 2.2 or a 3.3 kb fragment. The latter has been previously found to occur with significantly higher frequency in probands of families with familial hypoalpha. ApoA-I was isolated from three unrelated subjects with familial hypoalpha and the 3.3 kb PstI polymorphism of the apoA-I gene, and from normal control subjects. The apoA-I from the hypoalpha subjects was structurally normal as determined by amino acid analysis and by two-dimensional gel electrophoresis. When normal apoA-I and hypoalpha apoA-I were simultaneously injected into either normal controls or hypoalpha subjects, both forms of apoA-I were catabolized at the same rate in the same subject, indicating that the hypoalpha apoA-I is also metabolically normal. Analysis of the kinetics of metabolism of apoA-I in the hypoalpha subjects, compared to the normal controls, revealed that the reduced plasma levels of apoA-I were due to an increased apoA-I fractional catabolic rate, and that the synthetic rate was normal. Based on these results, we conclude that the apoA-I gene in these hypoalpha subjects is normal, and the PstI polymorphism near the apoA-I gene, which is associated with familial hypoalpha, is likely to be a marker for a mutant gene closely linked to, but not in, the apoA-I gene.     

Cholinesterase Levels in Blood Plasma and Erythrocytes from Calves,Normal Delivering Cows and Cows Suffering from Parturient Paresis

Plasma cholinesterase (pChE) levels and erythrocyte acetylcholinesterase (eAChE) levels were studied in 6 cows before, during and after parturition (Group I), their calves (Group II), 38 cows suffering from parturient paresis (Group III) and 14 newly delivered non-paretic cows (Group IV). The mean of the pChE level in Group I was 1.5 μkat/1 ± 0.20 before parturition and decreased significantly (P ≦ 0.05) to 1.2 ukat/1 ± 0.16 after parturition. The eAChE level was before parturition ≅ 140 ukat/1 and decreased to ≅ 130 μkat/1 4–5 weeks after parturition. At birth the pChE level was 12.8 ukat/1 ± 5.9 in Group II. After 4 weeks the level had decreased to 2.3 ukat/1 ±0.3. In the bull calves the pChE level started to increase when they were 6 weeks old and reached a level of 5.7 μkat/1 ± 0.6 before slaughter at 6 months of age. The heifers did not show this increase. They had a level of around 2 μkat/1 throughout the investigation. The eAChE level at birth was 119 μkat/1 and increased slowly to a level of 145 μkat/1 at 6 months. No differences between the sexes were found. The cows suffering from parturient paresis had a pChE level of 1.80 μkat/1 ± 0.30 before treatment with calcium (Ca). The level decreased significantly (P ≦ 0.001) after Ca-infusion to a level of 1.67 ukat/1 ±0.29. Group IV had a pChE level of 1.65 μkat/1 ± 0.42 at parturition. Two to 4 months later the cows that had recovered from milk fever had a level of 1.61 μkat/1 ± 0.31 and the control cows 1.66 ukat/1 ± 0.48. No differences between the groups were found for the eAChE level. The findings show that parturition influences the pChE level in cows and that sex influences the pChE level in calves between 6 weeks to at least 6 months of age. Furthermore the elevated pChE level found in the cows suffering from parturient paresis before Ca infusion may be a further sign of a disturbance in the cholinergic system with a special preference to the neuromuscular junctions.     

Immunolocalization of secretogranin II, chromogranin A, and chromogranin B in differentiating human neuroblastoma cells.

In order to obtain further insights into the expression of the known markers of secretory neuroendocrine dense core organelles, secretogranin II (SgII), chromogranin A (CgA), and chromogranin B (CgB) during neuronal differentiation, the immunolocalization of these proteins was studied by means of double immunofluorescence in both undifferentiated and retinoic acid-differentiated SH-SY5Y human neuroblastoma cells. The majority of undifferentiated cells was not immunolabeled for all three proteins. In the majority of differentiated cells, a clearly punctate SgII immunolabeling indicative of the presence of secretory organelles was present in the Golgi region, at the cell periphery, along the neurites and in growth cones. Only relatively few of the SgII-immunolabeled cells were also immunolabeled for CgA and CgB, and in a single cell the three proteins were not always present in the same organelles. These results, obtained in a cultured cell line, confirm the not necessarily parallel distribution of SgII, CgA, and CgB observed in different neuroendocrine tissues and suggest that SgII may be the best marker of human neuroblastoma cell differentiation.