Practice parameter: evaluation of the child with global developmental delay.
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Biomedical subjects
Publications and source records attributed to A Comi.
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Whole mount sections of the prostate are widely used in many laboratories. Macrocryosections of the gland; that is, whole mount frozen sections of the prostate from radical prostatectomies represent a useful new research protocol. The technique is very simple and does not require expensive equipment.
Copper incorporation (64Cu(II)) into Cu,Zn-superoxide dismutase (SOD) was studied in human lymphoblasts. Rapid incorporation of copper with a proportionate increase in SOD activity was detected. No copper incorporation or SOD activation was detected when 64Cu(II) was added to cell cytosols rather than to intact cells. Thus, incorporation of 64Cu was not due to isotopic exchange. Cycloheximide had no significant effect on copper incorporation and activation of SOD when the data were corrected for total cell copper. Thus, the data were consistent with copper incorporation into a preexisting apoSOD pool rather than newly synthesized SOD, and no new SOD synthesis was detected over a 15-h incubation period. The size of the apoSOD pool was estimated to be approximately 35% of the total SOD in lymphoblasts. When cells were preincubated for 15 h with excess copper (15 microM Cu(II)), the size of the apo pool markedly decreased but was not eliminated, suggesting that the apoSOD was not due to copper deficiency. These experiments also indicated that newly arrived copper was preferentially incorporated into the apoSOD pool, while the function(s) of an apoSOD pool remains unknown. Copper binding to apoSOD may provide a rapid protective response against copper toxicity.
The incorporation of copper into Cu,Zn-superoxide dismutase (SOD) was examined in Menkes lymphoblasts that express a genetic defect of copper metabolism. SOD activity was approximately 40% higher in Menkes than normal lymphoblasts. Since Menkes lymphoblasts contain elevated copper levels, the higher SOD activity is most likely due to near copper saturation of an apoSOD pool that is in normal lymphoblasts. Cycloheximide markedly inhibited 64Cu(II) incorporation into SOD in Menkes lymphoblasts under conditions in which no significant, de novo synthesis of SOD protein was detected with normal lymphoblasts. The maximal amount of 64Cu incorporation into newly synthesized SOD in Menkes lymphoblasts was approximately equal to the maximal amount of 64Cu that could be incorporated into the apoSOD pool in normal lymphoblasts. The increased synthesis of SOD in Menkes lymphoblasts may play a protective role against copper toxicity in Menkes lymphoblasts. The protonophore, CCCP markedly inhibited 64Cu incorporation into SOD in both normal and Menkes lymphoblasts, which is consistent with 64Cu incorporation into SOD within a membrane-bounded compartment in both cell types. When 64Cu-incorporation into SOD was blocked with CCCP, copper accumulated in a Superose column fraction that contains S-adenosylhomocysteine hydrolase (SAHH), which has a high affinity for copper. SAHH may play a role in delivering copper to SOD.
Proliferation and differentiation of villous trophoblast during placental development, from an early stage to full-term, were investigated in routinely fixed and processed tissues, by means of the immunocytochemical localization of the cell cycle-related proto-oncogene c-myc and the p53 and retinoblastoma susceptibility (Rb) tumour-suppressor gene products. The proliferative activity of the trophoblast was determined using an antibody against proliferating cell nuclear antigen (PCNA) which stains all proliferating cells in paraffin-embedded tissues. Diffuse nuclear immunoreactivity for PCNA, c-myc and Rb gene products was a consistent finding in early cytotrophoblast; c-myc product expression was also detectable in both layers of mid-gestation trophoblast. Only scattered cytotrophoblastic nuclei of early gestational placenta displayed immunostaining for p53 gene product. In full-term placenta c-myc expression was undetectable while Rb gene product and PCNA immunoreactivity declined markedly. These results indicate that the expression of the above genes is spatio-temporally regulated during placental development. A potential involvement of the oncosuppressor gene products p53 and Rb in the control of trophoblastic proliferation and of c-myc in the control of both the proliferative and differentiation pathways of trophoblastic cells is suggested.