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In a previous enzyme histochemical study, we showed that some of the uterine reserve cells carried heat stable alkaline phosphatase (ALP), but most of the cervical epidermoid cancers, heat sensitive ALP.From these result, we suspect that ALP isoenzyme pattern may change after malignant transformation (“enzyme deviation”). ALP reaction products in ALP positive cases, however, were not detected in all parts of the lesions but usually only in some parts.In this report, enzyme cytochemical two inhibition tests (heat stability and L-phenylalanine inhibition tests) were also applied to 55 cytological specimens from benign and malignant uterine tissues in order to find how often cell clusters of various of kinds of lesions have ALP activity and which type of ALP isoenzyme they have.The findings in this study may be summarized as follows:1. As in histochemical specimens, ALP activity was also found in cytological specimens of reserve cell, reserve cell hyperplasia after cryosurgery, cervical gland, cervical epidermoid cancer, endometrium in the proliferative phase and endometrial adenocarcinoma.2. In the heat stability test (65°C, 30min.) for cytological specimens, ALP of reserve cell, reserve cell hyperplasia after cryosurgery and cervical gland were heat stable. ALPs of endometrium and epidermoid cancer, however, were heat sensitive. Of 18 cases of endometrial cancer, the ALPs from six were heat stable, and the others heat sensitive.3. In the L-phenylalanine inhibition test, ALPs from all lesions were sensitive to a lesser or greater extent.From these cytochemical studies of ALP, it may be postulated that “enzyme deviation” exists in the course of carcinogenesis of both uterine cervical epidermoid cancer and endometrial adenocarcinoma.

The Regan isoenzyme, a heat-stable alkaline phosphatase, which has been thought to be derived only from cancer or the placenta, has been reported in non-cancer, non-pregnant patients. We performed histochemical studies to determine whether the heat-stable alkaline phosphatase exists in normal cells of the female reproductive organ. For this purpose, the heat-stability test and the L-phenylalanine inhibition test were used. Heat-stable and L-phenylalanine-sensitive alkaline phosphatase was found in normal cells of Muellerian duct origin, e.g., uterine cervical reserve cells, cervical columnar cells and epithelial cells of the endometrial surface. Our findings suggest that heat-stable alkaline phosphatase, such as the Regan enzyme, may be produced by normal cells of Muellerian duct origin.

Tomato cultivars Zuiko (ZK), resistant to TMV causing top-necrosis by infection with TMV, Fukuju No. 2 (F2), susceptible to TMV causing mosaic, strains GCR 236, GCR 237, GCR 254, and GCR 267, resistant to TMV causing no symptoms, were used. They were tested whether or not the induced resistance does occur by changing temperatures during incubation following inoculation, by heating tomato seedlings as a shock at 45°C for 10 min immediately after inoculation, by introducing dyes, acridine orange (AO), azure B (AB), and pyronine B (PB), into tomato seedlings, and by introducing the infected tomato extract.At 25°C, TMV concentration in ZK was 2% of that in F2, whereas it reached almost 60% at 20°C, showing that ZK was not resistant at a low temperature below the optimum for tomato growth. Heat shock induced the reduced amount of TMV in ZK as compared with non-treatment, but did not in GCR 236, GCR 237, and GCR 267. Thus, ZK alone showed an induced resistance by this treatment. ZK, GCR 236, GCR 237, and GCR 254, all showed a decrease in TMV concentration by introducing dyes through root, especially by 10 ppm AO. However, F2 cultured in 10 ppm AO showed no alteration in TMV concentration comparing with that cultured in water. When the extract obtained from non-infected ZK which was cultured in 10 ppm AO, was mixed with TMV and the mixture was inoculated on the leaves of bean (Phaseolus vulgaris), AO absorbed-ZK extract showed no inhibitory activity to TMV infection. These facts indicate that AO did not directly inactivate TMV, but induced a highly resistance in ZK except F2. F2 which had absorbed the infected F2 extract, reduced TMV concentration, but that absorbed non-infected F2 extract or infected ZK extract did not so much. ZK did not reduce TMV concentration by absorbing F2 extract or non-infected ZK extract, but did a little by absorbing infected ZK extract.From these results, induction of resistance in tomato cultivars caused by the environmental factors is discussed.

We report a histochemical study of alkaline phosphatase (ALP) in normal cells of the female reproductive system, in pre-cancerous and cancerous lesions of the uterine cervix and in endometrial cancer to ascertain the incidence of ALP and its isoenzyme type. For this purpose, serial sections were subjected to heat stability and L-phenylalanine (LP) inhibition tests. The Regan-like isoenzyme, a heat-stable and LP-sensitive ALP, which has been thought to derive only from cancer or the placenta, was found in uterine cervical reserve cells and endometrial luminal surface lining cells. In contrast, ALP activity in endometrial glandular cells was found to be heat and LP sensitive. Of 183 cases of cervical neoplasia, 60 (33%) manifested non-specific ALP activity. One dysplasia and two invasive cancer cases manifested the Regan-like isoenzyme. The other 36 classifiable lesions had small-intestine ALP-like activity (marked heat and LP sensitivity) or a liver ALP-like isoenzyme (marked heat and slight LP sensitivity). Of 42 cases of endometrial cancer, all cases manifested non-specific ALP activity. Seven endometrial cancers exhibited the Regan-like isoenzyme. The other 19 cases manifested either small intestine or liver ALP-like isoenzyme. Our findings indicate that in the course of uterine carinogenesis, the ALP isoenzyme of reserve cell and endometrial glandular cells undergo a change and that enzyme deviation occurs.