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(7), we provide additional evidence on the distribution of the multiple (up to 16) phosphorylation sites within the native kinase and show that the 35-kDa COOH-terminal fragment (F35) released by trypsin digestion has full catalytic activity and two to three of the phosphorylation sites of native kinase. In this paper, we report the cloning and sequencing of the corresponding cDNA and expression of a fully active F35. MATERIALS AND METHODScDNA Cloning-Total RNA was purified from Acanthamoeba castellanii by single step thiocyanate-phenol-chloroform extraction (Stratagene RNA isolation kit) and mRNA was purified using Poly-T columns (Stratagene). cDNA corresponding to the F25 fragment of MIHC kinase (7) was prepared by the 3Ј-rapid amplification of cDNA ends method using Life Technologies, Inc. reagents and following the company's protocol using poly(T) primer for reverse transcription. The degenerate primers used for PCR reactions corresponded to internal portions of the NH 2 -terminal sequences of the F35 and F11 tryptic fragments of MIHC kinase (

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Cross-sectional studies in France have shown strong regional correlations between death rates from alcohol related diseases and death rates from gastric cancer. The present study involved 40 cases of newly diagnosed adenocarcinoma of the stomach and 168 control subjects with one of four other gastrointestinal diagnoses selected from the same hospital service during the same time period, 1978-1980. On the basis of a standard nutritional interview alcohol and particularly red wine were seen to be significant risk factors for this cancer (relative risks of 6.9 with 95% confidence limits (CL) of 3.3-14.3 for alcohol and 6.3 with CL 3.1-12.7 for wine). Smoking of one or more cigarettes per day was associated with a relative risk for gastric cancer of 4.8 with CL of 1.6-14.8. The presence of both risk factors was associated with a relative risk of 9.3 with 95% CL of 4.6-19.0. Possible confounding by age, smoking, and eating lettuce (a reported protective factor for gastric cancer in other studies) did not explain these results. The relative risks were consistently found and remained significant when each diagnostic group of control subjects was analyzed separately. These results suggest that alcohol, and particularly red wine, may be important risk factors for adenocarcinoma of the stomach in France. In addition, cigarette smoking, a risk factor in itself, when coupled with alcohol appears markedly to increase the risk.