The Non-genotoxic Carcinogen Study Group in the Environmental Mutagen Society of Japan organised the second step of the inter-laboratory collaborative study on one-stage and two-stage cell ...transformation assays employing BALB/c 3T3 cells, with the objective of confirming whether the respective laboratories could independently produce results relevant to initiation or promotion. The method was modified to use a medium consisting of DMEM/F12 supplemented with 2% fetal bovine serum and a mixture of insulin, transferrin, ethanolamine and sodium selenite, at the stationary phase of cell growth. Seventeen laboratories collaborated in this study, and each chemical was tested by three to five laboratories. Comparison between the one-stage and two-stage assays revealed that the latter method would be beneficial in the screening of chemicals. In the test for initiating activity with the two-stage assay (post-treated with 0.1μg/ml 12- O-tetradecanoylphorbol-13-acetate), the relevant test laboratories all obtained positive results for benzo apyrene and methylmethane sulphonate, and negative results for phenanthrene. Of those laboratories assigned phenacetin for the initiation phase, two returned positive results and two returned negative results, where the latter laboratories tested up to one dose lower than the maximum dose used by the former laboratories. In the exploration of promoting activity with the two-stage assay (pretreated with 0.2μg/ml 3-methylcholanthrene), the relevant test laboratories obtained positive results for mezerein, sodium orthovanadate and TGF-β1, and negative results for anthralin, phenacetin and phorbol. Two results returned for phorbol 12,13-didecanoate were positive, but one result was negative — again, the maximum dose to achieve the latter result was lower than that which produced the former results. These results suggest that this modified assay method is relevant, reproducible and transferable, provided that dosing issues, such as the determination of the maximum dose, are adequately considered. The application of this two-stage assay for screening the initiating and promoting potential of chemicals is recommended for consideration by other research groups and regulatory authorities.
Emotional sweating is a physical reaction that occurs with pain and other acutely stressful situations. Very few studies have directly evaluated emotional sweating to monitor pain reception in ...full–term newborns. The aim of study was to examine whether emotional sweating could applicate for evaluating procedural pain by heel lance in full–term newborns. Eight full–term newborns participated on the fourth day after birth in this study. We examined whether the amount of sweat secretion changed during blood collection procedure. The sweating reaction was recorded continuously from the start of the blood collection until blood collection was finished, using the probe of a portable perspiration meter against the newborn’s palm. As a result, the amount of emotional sweat significantly increased in perspiration accompanied the heel lance, compared to the baseline before blood collection. These finding suggest that emotional sweating could be used as an objective index of procedural pain in full–term newborns.
We previously showed that fatty liver was easily induced in suncus by starvation and that the plasma level of apolipoprotein B (apo B) was very low. There are three possible explanations for the low ...level of apo B in the animals: low synthetic rate, low secretion rate, and rapid catabolism in the circulation of apo B. We measured post-heparin lipolytic activity (lipoprotein lipase activity), which plays a key role in the catabolism of apo B–containing lipoprotein, VLDL, and found no difference between rats and suncus. We also investigated the hepatic synthetic rate of apo B by liver perfusion studies. Newly synthesized apo B in the suncus liver was detected by immunoprecipitation and found to amount to 12.5% of that in rats. The secretion rate of VLDL in suncus, which was estimated by intravenous injection of Triton WR1339, was 13.8% of that in rats. These two results suggest that there is no major defect in the secretory process. We separated Golgi apparatus from rat and suncus livers, and found much fewer lipoprotein particles in suncus than in rat Golgi apparatus. This evidence suggests that there is no defect in the lipolytic process or hepatic secretory process of apo B–containing lipoprotein, VLDL, but there may be a defect in the assembly process of VLDL and/or in the synthetic process of apo B in suncus. Such a defect may be one of the reasons for starvation-induced fatty liver in suncus
The author compared two groups of female patients with insert dysfunction whom he treated. One group is the patients who were treated from 1986 to 1990 (the 90 group) and the other is those treated ...from 1996 to 2000 (the 2000 group). Having compared the two groups, the following five important findings became clear. Compared with the 90 group, the patients of the 2000 group show that they : 1. Take more time before they come to hospital for help. 2. Need more time for treatment. 3. Have more difficulty in starting psychotherapy, and in fostering their understanding during the treatment. 4. Trend to flight into pregnancy. 5. Trend to choose partners whose problems are obvious from the beginnings. This tendency of not comforting the problems is not limited to female patients with insert dysfunctions, but common to male and female patients with other sexual dysfunctions. Furthermore we find similar tendencies in the patients suffering from problems with children or adolescents.
We have previously shown that fatty liver was easily induced in suncus by starvation and that the plasma level of apolipoprotein B (apoB) was very low. We also previously reported that a defect in ...the assembling process of apo B-containing lipoprotein (very low density lipoprotein, VLDL) may be one of the reasons for the low level of plasma apo B and for induction of fatty liver by starvation in suncus. We also found that hepatic acyI coenzyme A cholesterol acyltransferase (ACAT) activity is very low in the animals, resulting in decreased cholesteryl ester contents in the liver. A deficiency of cholestery1 ester in suncus liver may be one of the reasons for the defect in the assembling process of VLDL. In this study, we investigated the effect of cholesterol-feeding, which induces an increase in triglyceride and cholestery1 ester of the liver as a consequence of the induction of both intestinal and hepatic ACAT activities, on the secretion of VLDL. Although the basal ACAT activity of intestinal mucosa was high, cholesterol-feeding did not induce either an increase in plasma lipid or an increase in intestinal ACAT activities in suncus. The hepatic secretion rate of VLDL was estimated by treatment with Triton WR1339, which is well known to inhibit the catabolism of VLDL. Cholesterol-feeding caused a slight increase in hepatic triglyceride and cholesteryl ester but no increase either in the secretion rate of VLDL or in hepatic ACAT activity. This evidence suggests that either the secretion rate of VLDL is not dependent on absolute hepatic contents of cholesteryl ester, or it is dependent on the level of cholesteryl ester which is produced by hepatic ACAT.
The Non-genotoxic Carcinogen Study Group in the Environmental Mutagen Society of Japan organised the second step of the inter-laboratory collaborative study on one-stage and two-stage cell ...transformation assays employing BALB/c 3T3 cells, with the objective of confirming whether the respective laboratories could independently produce results relevant to initiation or promotion. The method was modified to use a medium consisting of DMEM/F12 supplemented with 2% fetal bovine serum and a mixture of insulin, transferrin, ethanolamine and sodium selenite, at the stationary phase of cell growth. Seventeen laboratories collaborated in this study, and each chemical was tested by three to five laboratories. Comparison between the one-stage and two-stage assays revealed that the latter method would be beneficial in the screening of chemicals. In the test for initiating activity with the two-stage assay (post-treated with 0.1μg/ml 12-O-tetradecanoylphorbol-13-acetate), the relevant test laboratories all obtained positive results for benzoapyrene and methylmethane sulphonate, and negative results for phenanthrene. Of those laboratories assigned phenacetin for the initiation phase, two returned positive results and two returned negative results, where the latter laboratories tested up to one dose lower than the maximum dose used by the former laboratories. In the exploration of promoting activity with the twostage assay (pretreated with 0.2μg/ml 3-methylcholanthrene), the relevant test laboratories obtained positive results for mezerein, sodium orthovanadate and TGF-β1, and negative results for anthralin, phenacetin and phorbol. Two results returned for phorbol 12,13-didecanoate were positive, but one result was negative — again, the maximum dose to achieve the latter result was lower than that which produced the former results. These results suggest that this modified assay method is relevant, reproducible and transferable, provided that dosing issues, such as the determination of the maximum dose, are adequately considered. The application of this two-stage assay for screening the initiating and promoting potential of chemicals is recommended for consideration by other research groups and regulatory authorities.
The body size and age structure of a breeding population of the Japanese common toad, Bufo japonicus formosus, was studied at Yamakitamachi in Kanagawa Prefecture for three breeding seasons from 1992 ...to 1994. Yearly variation in snout-vent length (SVL) was not significant, but the difference in SVL between the sexes was highly significant; the mean SVL of males and females was 125.8 and 134.2 mm, respectively. The breeding adults were aged successfully by skeletochronology using phalanges. The breeding adults were 1-8 yr old. The mean and median ages were 4.5 and 4 yr in males, and 5.2 and 5 yr in females, respectively. The males were estimated to begin to breed when one to four yr old, with a mean of 2.4 yr, while the females were estimated to breed when two to four yr old, with a mean of 2.7 yr. Age at first reproduction was significantly younger for males than females. We discuss the sexual size dimorphism of B. j. formosus in relation to the difference in age structure of the breeding population between the sexes clarified by skeletochronology.
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