Characteristics of cis-diamminedichloroplatinum-selected in vitro passaged cells derived from a transplantable rat malignant fibrous histiocytoma

A cis-diamminedichloroplatinum (CDDP)-selected cell line (MT-R10) was induced by continuous exposure of an in vitro passaged cell line (MT-P) established from a transplantable rat malignant fibrous histiocytoma (MFH-MT) to CDDP. MT-R10, capable of proliferating in the presence of 1.0 microgram CDDP/ml, was passaged in CDDP-free medium. The doubling time of MT-R10 at passage 10 (MT-R10/10) was almost the same as that of MT-P, being 22.3 and 25.5 h, respectively. The concentration of CDDP required for 50% inhibition of MT-R10/10 proliferation was two-fold higher than that of MT-P. MT-R10 consisted of round, epithelial-type cells arranged in compact sheets. Ultrastructurally, MT-R10 had numerous free ribosomes, some mitochondria, and other poorly developed cytoplasmic organelles suggesting its undifferentiated nature. MT-R10 showed no reaction for acid phosphatase or non-specific esterase. Tumors induced in syngeneic rats by inoculation with MT-R10 consisted of small, round, undifferentiated cells with scanty cytoplasm. They showed organoid and trabecular patterns, and were often arranged in compact sheets. The neoplastic cells showed no reaction for any of the histiocytic lysosomal and antigenic markers tested, but exhibited a strong reaction for alkaline phosphatase. Bone formation was often observed in the tumors. These observations suggest that CDDP-selected, undifferentiated cells may have osteogenic potential and may be one of the progenitor cells of MFH-MT.     

CORRELATION BETWEEN GLYCOCONJUGATE EXPRESSION AND FUCOSYLTRANSFERASE ACTIVITY IN HUMAN COLORECTAL CARCINOMA

Using the surgically extirpated specimens from 9 patients with colorectal carcinoma, fucosyltransferase activities in the carcinoma tissue and the normal mucosa were measured and were compared with the hlstochemical findings of glycoconjugates which were shown by staining with lectins reacting with blood group antigens and related substances. The fucosyltransferase activities of the carcinoma tissue were well correlated with the overall findings of lectin stainings after neuraminidase treatment. The more intense the carcinoma tissue was stained, the higher the fucosyltransferase activity was shown. However, there were marked differences in the fucosyltransferase activities by the portions measured, depending upon the relative amount of carcinoma tissue and Interstitial tissue; in the invasive portion with less carcinoma tissue, the activity was generally low in comparison with that in the surface area where carcinoma tissue was rather abundant. Thus, the morphological and lectin hlstochemical finding are of paramount importance for the eveluation of glycosyltransferase activity in human colorectal carcinoma.     

A new method for preventing pulse pileup in scintillation detectors

A new method for preventing pulse pileup in scintillation detectors is proposed. In the new method (G-INT), the energy of an event is calculated from the 'gated integral' of the pulse signal and the period of integration. The period of integration is not fixed but is shortened by the arrival of the succeeding pulse so as to avoid post-pulse pileup. The effect of pre-pulse pileup is corrected by subtracting the remnant energy of the preceding pulses, which is calculated from the gated integral of the preceding pulse. To avoid error due to short pulse intervals, pre- and post-pulse deadtimes are imposed. The method is similar to Wong's method (W-SUM) that depicts the energy by the 'weighted sum' of the current signal and the integrated signal. The performance of G-INT has been studied by Monte Carlo simulation in comparison with W-SUM, the variable sampling-time technique and simple delay-line clipping. It is shown that G-INT provides the smallest degradation in pulse height resolution for a given count rate capability. The difference between G-INT and W-SUM is explained by the difference in the amount of statistical noise involved in the gated integral and in the weighted sum.     

Twice-daily measurements of stature and body weight in two children and one adult

Stature and body weight of two children, 12.5 and 9.3 years, and their mother were measured twice daily for 488 days, immediately after rising and just before bed. The time of the measurements was also recorded and diurnal changes during daytime and nighttime were estimated. Stature began to decrease instantaneously after rising. Therefore, morning measurements were made immediately after rising. Stature decreased during the day, and the mean daytime loss was 1.78 cm in the older child, 1.61 cm in the younger child, and 1.43 cm in the adult. Stature increased during sleep hours at night, and the mean nighttime gain was 1.79 cm, 1.63 cm, and 1.43 cm in each subject, respectively. Stature also increased after naps and a bath. Body weight decreased during sleep hours at night, and the mean nighttime loss was 0.46 kg, and 0.38 kg, and 0.36 kg in each subject, respectively. Diurnal changes were largest in the older child, while the ratios of changes (ΔST/ST, ΔWT/WT) were largest in the younger child whose growth rate was the greatest. In the children, the nighttime gain of stature was not as highly correlated with hours of sleep as in the adult. Diurnal change varies depending upon sleep hours and stage of growth. Rapid growth in stature of 1 cm in only three weeks was observed once in the younger child. During this period, diurnal change was smaller than the mean value over 488 days. In the older child, the growth rate decreased in both stature and weight about three months after menarche. © 1993 Wiley-Liss, Inc.     

Clinical importance of serum anti-p53 antibodies as tumor markers

Anti-p53 antibodies are autoantibodies induced by mutation of p53 cancer-suppressor gene, and are considered to be indirect markers for p53 gene mutations and abnormally high p53 gene levels. We evaluated the usefulness of the measurement of anti-p53 antibodies by enzymed-linked immunosorbent assay using serum samples from patients with various disorders and normal subjects. The anti-p53 antibody concentration was high in patients with lung, esophageal, gastric, hepatocellular, colonic, rectal or ovarian cancer and significantly differed between the group with neoplasms and those with non-neoplastic disorders. Particularly high concentrations were observed in patients with malignant tumors. The mean agreement rate between anti-p53 antibodies and conventional tumor markers was only 47.8% despite slight differences among disorders. The positive rate increased to 63.0% by their combination assay. In addition, anti-p53 antibodies were independent markers, not complimentary to conventional markers. The mean agreement rate between anti-p53 antibodies and tissue p53 was 70.0%. Though the anti-p53 antibody-positive rate was lower than the tissue p53-positive rate, anti-p53 antibodies may be useful new tumor markers because specimens from the affected tissue are not necessary.     

Neurite extension of DRG neurons by gicerin expression is enhanced by nerve growth factor

Gicerin, a cell adhesion molecule, is expressed in dorsal root ganglion (DRG) and sciatic nerves during chick development. This molecule re-appears in these tissues after an injury to the sciatic nerve. In the present study, we investigated the expression of nerve growth factor (NGF) in the regenerating sciatic nerve of chicks and the effects of NGF on the expression and neurite activities of gicerin in DRG. In the sciatic nerve after a crush injury, the expression of NGF and gicerin increased in the Schwann cells and in the nerve fibers, respectively. NGF promoted the neurite projections from in vitro DRG on the gicerin ligands, which were inhibited by anti-NGF antibody. The gicerin mRNA expression increased in the DRG with NGF, which was inhibited by the co-incubation with anti-NGF antibody. These results indicate that NGF might therefore enhance the expression of gicerin in DRG, thereby promoting the gicerin-dependent neurite extension during sciatic nerve regeneration.     

Hydrolysis of Carbonates,Thiocarbonates, Carbamates,and Carboxylic Esters of α-Naphthol, β-Naphthol,and p-Nitrophenol by Human,Rat, and Mouse Liver Carboxylesterases

Thirty carbonates, thiocarbonates, carbamates, and carboxylic esters of -naphthol, -naphthol, and p-nitrophenol were synthesized and tested as substrates for liver carboxylesterases from the crude microsomal fractions of human and mouse, and purified isozymes, hydrolases A and B, from rat liver microsomes. The carbonates, thiocarbonates, and carboxylic esters of -naphthol were cleaved more rapidly than the corresponding -naphthol isomers by the mammalian liver esterases. -Naphthyl esters of acetic, propionic, and butyric acids were among the best substrates tested for these enzymes. The majority of the substrates was consistently hydrolyzed at higher rates by hydrolase B compared with hydrolase A, although the Michaelis–Menten constant (K _m) values of selected substrates differed widely with these two isozymes. Malathion was a 15-fold better substrate for hydrolase B than for hydrolase A. Compared with the corresponding carboxylates, the carbonate moiety of - and -naphthol and p-nitrophenol lowered the specific activities of the enzymes by about fivefold but improved stability under basic conditions. The optimum pH of mouse liver esterase with the acetate, methylcarbonate, and ethylthiocarbonate of -naphthol was between pH 7.0 and pH 7.6. Human and mouse liver microsomal esterase activities were about five orders of magnitude lower than the esterase activities of purified rat liver hydrolase B. A relationship between the catalytic activity of the enzymes and the lipophilicity of the naphthyl substrates indicated that (i) in the - and -naphthyl carbonate series, an inverse relationship between enzyme activity and lipophilicity of the substrates was observed, whereas (ii) in the -naphthyl carboxylate series, an increase in enzyme activity with increasing lipophilicity of the substrates up to a log P value of about 4.0 was observed, after which the enzyme activity decreased.     

A comparative study of measurement of arterial blood pressure using HEM-802F and arterial cannulation

Finger arterial blood pressures determined by a newly developed sphygmomanometer, HEM-802F, were compared with arterial pressure obtained from direct measurement of the radial artery. An excellent correlation was found between the two methods (systolic: r = 0.93, diastolic: r = 0.91), although there was a large variability among individual subjects. The range of differences between them are from +32 to –13mmHg for systolic and +15 to –25mmHg for diastolic blood pressure measurement. HEM-802F underestimated systolic pressure (–4.0mmHg) and overestimated diastolic pressure (+6.7mmHg), compared with intra-arterial readings.The HEM-802F was useful for the non-invasive arterial pressure monitoring during general anesthesia.(Shigemi K, Takahashi H, Hashimoto S et al.: A comparative study of measurement of arterial blood pressure using HEM-802F and arterial cannulation. J Anesth 4: 91–93, 1990)     

Suppression of neointimal thickening by a newly developed HMG-CoA reductase inhibitor,BAYw6228, and its inhibitory effect on vascular smooth muscle cell growth

1. The aim of this study was to determine whether BAYw6228 (BAYw), a newly developed 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase inhibitor, could suppress an atherogenic process such as intimal thickening by a mechanism other than lowering the level of serum cholesterol.
2. First, we evaluated the in vitro effect of BAYw on the proliferation of vascular smooth muscle cells (SMC) from various species: Sprague-Dawley (SD) rats, New Zealand (NZ) white rabbits, intimal cells from Watanabe hereditary hyperlipidemic (WHHL) rabbit and SMC from the new-born human aorta. The increasing rate of total protein content of these cells was inhibited by the addition of BAYw in a dose-dependent fashion. In the presence of 2% foetal calf serum (FCS), the value of IC₅₀ was 1.0 μM in SD rats. 2.1 μM in NZ white rabbits, and 0.3 μM in WHHL rabbits. With human SMC, the value was 0.02 μM in the presence of 10% FCS and 0.2 μM with a mixture of growth factors.
3. Based on these above in vitro findings, we next examined the in vivo effect of the agent to determine whether it could suppress rabbit intimal thickening induced by balloon catheterization. A balloon catheter was inserted from a peripheral branch of the left external carotid artery to the aorta to denude the endothelium of the left common carotid artery in Japanese white rabbits. After 12 days they were divided into control and BAYw groups. The former were subcutaneously injected with saline and the latter with BAYw 1 mg kg⁻¹ day⁻¹. Two days after the beginning of treatment, a second balloon injury was performed to the previously injured left common carotid artery in both groups. After another two weeks, the left common carotid artery was removed and variously stained. Although the total serum cholesterol in the BAYw group was significantly lower than in the control (P<0.05), the difference was not enough to affect intimal thickening. In addition, the BAYw group had a smaller intima/media ratio than the control group, decreasing to 45% of control (P<0.05). By anti-α smooth muscle actin antibody staining, these intimal thickening areas were entirely occupied by SMCs, and their amount was attenuated by BAYw. By anti-rabbit macrophage antibody (RAM 11) staining, the number of positive cells in the intimal thickening was markedly decreased in the BAYw group compared to control (P<0.01).
4. These results indicate that BAYw has an inhibitory effect on intimal thickening by attenuating intimal SMC proliferation and infiltration of macrophages, suggesting that BAYw could be effective in the prevention of the progression of atherosclerotic plaque-like restenosis after angioplasty.