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The nucleolar and mitochondrial morphology of developing reconstructed bovine nuclear transfer (NT) embryos and stage-matched in vivo-produced control embryos were examined under the electron microscope. Each reconstructed embryo at the one-cell (n = 12), two-cell (n = 5), three-cell (n = 3), four-cell (n = 5), 5-8-cell (n = 5) and blastocyst (n = 3) stages was produced by fusion of a 16-32-cell-stage blatomere with an aged enucleated bovine oocyte. The normal and reconstructed embryos showed similar mitochondrial morphology. However, NT embryos produced several pleiomorphic forms not seen in controls, and were more heterogeneous at early stages of development. Control embryos exhibited nucleolar features considered indicative of rRNA synthesis from the eight-cell stage onwards. In contrast, the NT embryos presented nucleoli with morphology consistent with rRNA synthesis in all embryos examined, except in the three-cell and in two of the five four-cell embryos. From this nucleolar morphology, it was concluded that nuclear reprogramming does not occur immediately following nuclear transfer, but occurs gradually over the first two or three cell cycles.

Purpose: Many small (less than 0.5 cc), well differentiated, organ-confined prostate carcinomas remain clinically undetected during the life of the patient and are identified only at postmortem examination. Thus, these cancers are often called latent or autopsy cancers. There is concern that serum prostate specific antigen (PSA) based screening may preferentially detect these cancers. There are limited prospective data concerning the pathological features of carcinomas of the prostate detected in a screening program. We determined if prostatic carcinomas detected via PSA based screening resembled autopsy cancers., Materials and Methods: We assessed the pathological features of carcinomas in 100 consecutive, completely embedded radical prostatectomy specimens from men whose cancer was detected in a PSA based screening program. The tumors were evaluated for pathological stage, surgical margin status, Gleason histological grade and intraglandular tumor extent (morphometrically quantified as percentage carcinoma and tumor volume)., Results: Of 100 carcinomas 68 (68%) were larger than 0.5 cc in volume (mean 1.7, range 0.1 to 10.7). Mean amount of carcinoma in the surgical specimen was 10.3% (range 0.1 to 41.6). Of the 100 carcinomas 94 had a Gleason score of 5 to 8 (mean 5.7) and only 6 (6%) were well differentiated (Gleason score of 4 or less). Locally advanced disease was noted in 41 cases (41%) as judged by the presence of extracapsular carcinoma and/or cancerous surgical margins., Conclusions: We concluded that the pathological features of most prostatic carcinomas detected via PSA based screening do not resemble those of autopsy cancers, and that most prostatic cancers detected in screening programs are likely to be clinically important.

The timing of genome activation in bovine embryos is still not well defined. The objective of this study was therefore to investigate transcription in bovine embryos with a high potential to develop in culture after in vitro fertilization, by examining, autoradiographically, their incorporation of 3H-uridine. Initial experiments determined that developmental potential in vitro could be related to the time of first division of the zygote. Embryos that completed their first cleavage within 30 hours of exposure to sperm were more likely to develop into blastocysts (65.7%) and to hatch (50.9%). Using such embryos, it was found that 10 of 12 8-cell and all 11 4-cell stage embryos were labeled after a 2-4-hr exposure to 3H-Uridine. Among 2-cell stage embryos, 0 of 23, 3 of 17, 8 of 15, and 3 of 4 were labeled after exposure to 3H-uridine of 2, 4, 7, and 10 hr, respectively. Treatment with alpha-amanatin (10-100 micrograms/ml) blocked 3H-uridine incorporation but did not inhibit cleavage during the first 4 cell cycles. It was concluded that transcription occurs as early as the 2-cell stage in bovine embryos in vitro but is not critical to the first four cell cycles.