All cancer staging systems seek to identify clinical and pathological features that can predict outcome or guide therapy. In particular, a non‐invasive method for the early detection of disseminating disease would be of great interest. We investigated the use of cytokeratin genes expression to detect blood metastases from colorectal tumors. Epithelial tumor cells were isolated from whole blood using the monoclonal antibody (MAb) BerEP4 and magnetic beads, and detected by reverse transcription‐polymerase chain reaction using oligonucleotides derived from the cDNA sequences of cytokeratins 8, 19 and 20. The sensitivity of this assay was determined by spiking SW620 colon carcinoma cells in normal blood. Using cytokeratin 19 expression we were able to detect 1 epithelial tumor cell in 1 ml of whole blood. The clinical applicability of this technique was explored by evaluating patients with a colorectal carcinoma. Epithelial cells were detected in the blood of 12 out of 23 patients, 2 (20%) of 10 with Astler‐Coller stage A or B, and 10 (77%) of 13 with stage C or D cancer. In conclusion, this test is a non‐invasive, sensitive, and specific assay for detecting circulating epithelial cells in blood. It may be useful for the early diagnosis of disseminating disease, to determine whether the presence of micrometastatic cells at the time of surgery is correlated with an early relapse and for monitoring adjuvant therapeutic trials. Int. J. Cancer 74:540–544, 1997. © 1997 Wiley‐Liss, Inc.

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A utomated cell culture incubators generally are considered primary components of fully automated cell culture systems, which are able to monitor cell growth without human interaction. This tutorial is focused on automated cell culture incubators. It emphasizes the impact of automation on throughput and environmental controls (temperature, humidity, and CO 2) and proposes some basic protocols to check these functions. In addition, it details practical aspects for switching from manual to automated cell culture incubators.

The aim of this work was to investigate the presence of inactive renin (IR) in plasma of normal infants and children and nephrectomized children and to study the plasma IR response to stimulation of the renin-angiotensin system (orthostasis) in children. The study was performed in 10 normal infants (2 days to 1 yr old), 28 normal children (1-15 yr old), 8 nephrectomized children (8-14 yr old), and 7 normal adults (20-40 yr old). IR was calculated as the difference in renin activity in trypsin-treated (1500 micrograms/ml) plasma, e.g. total renin (TR), and in untreated plasma, e.g. active renin (AR). IR was not detectable in most infants in the supine position, but their AR values were high (8.8-30 ng/ml X h). Moreover, in some of these infants, trypsin appeared to degrade renin activity, since TR values were lower than AR values. IR was detectable in 3 infants and 27 children, but their AR values were in a lower range (0.3-10 ng/ml X h). Trypsin degradation of renin activity was not found in either children or adults. With increasing age (2 days to 40 yr), AR decreased while IR and the IR to TR ratio increased significantly (P less than 0.001). A significant (P less than 0.001) inverse relationship was found between the IR and AR values of subjects 2 days to 40 yr old. IR was detectable in all nephrectomized children and represented 25% of normal values, while AR was undetectable (less than 0.1 ng/ml X h). In children in the upright position, IR decreased and AR increased significantly (P less than 0.001) in a reciprocal manner. TR did not change. These data suggest 1) that trypsin degradation of renin activity and absence of trypsin-activated IR are specific to infants with high AR levels, and 2) that IR might be activated in vivo into AR, especially after changes in position in children. IR could be a prorenin playing a physiological role in children.