Protein crystallization in a 100 nl solution with new stirring equipment.

To investigate quantitatively the effects of stirring on protein crystallization, a new stirring system which can agitate a protein solution, approximately 100 nl, by providing Hagen-Poiseuille flow has been successfully developed. In addition, this new stirring system provides flow with a well defined pattern and velocity. Using this system, hen egg-white lysozyme was crystallized in 100-200 nl solutions while being stirred. The optimum stirring conditions for lysozyme crystals have been explored by evaluating the Reynolds (Re) number and the crystals obtained. Intermittent flow, as well as a low Re number, was found to contribute significantly to the growth of a smaller number of larger crystals.

Different effects of methylxanthines on central serotonergic postsynaptic neurons in a mouse behavioral model.

Effects of the four methylxanthines (100 mg/kg, IP)--caffeine, theophylline, theobromine, and pentoxifylline--on the central serotonergic neuron were studied in mice using a behavioral model, the head-twitch response. The four methylxanthines potentiated the head twitches induced by 5-hydroxytryptophan (5-HTP) in pargyline-pretreated mice; pentoxifylline was the most potent. The potentiating effect of pentoxifylline was increased by paroxetine, the selective inhibitor of uptake of 5-hydroxytryptamine (5-HT), but those of the other drugs were not. In nontreated animals, caffeine directly induced head-twitch responses, which were not affected by pargyline pretreatment but were increased by prior treatment with 5,7-dihydroxytryptamine (5,7-DHT). The number of head twitches produced by caffeine in 5,7-DHT-treated mice was increased twofold by p-chlorophenylalanine (p-CPA), the tryptophan hydroxylase inhibitor. In mice treated with both 5,7-DHT and p-CPA, theophylline induced the responses, although much less potently than caffeine. Theobromine and pentoxifylline produced even fewer responses. From the results of the present study, it may be concluded that the methylxanthines possess qualitatively different actions on the central serotonergic neuron; caffeine and theophylline appear to have direct effects on the postsynaptic neuron, but theobromine and pentoxifylline do not.

[Fertility study of cefodizime sodium in mice--intravenous administration from pre-conceptional period through early period of gestation].

Cefodizime sodium (THR-221) was intravenously administered to mice of both sexes at dose levels of 300, 1000, 2000 and 3000 mg/kg/day, and its effects on the fertility of male and female mice and on their offspring were examined. Twenty-four males and 24 females were used per dose and as a control group. The males were dosed for 9 weeks prior to mating and through the mating period, and the females were treated from 2 weeks before mating through day 6 of gestation. Cesarean sections were performed on all pregnant mice in each group (16-23) on day 18 of gestation. Of the parental mice, 1, 2 and 1 males died at 0 (control), 2000 and 3000 mg/kg/day, respectively, and 5 females died at 3000 mg/kg/day. Inflammatory changes in the skin at the injection site were observed in the males at 3000 mg/kg/day. In the males, the kidney weight at 3000 mg/kg/day and the spleen weights at 2000 and 3000 mg/kg/day increased, and the thymus weight at 3000 mg/kg/day decreased, as compared with the control values. No differences appeared between any compound-treated group and the control group in the following maternal and fetal parameters: copulation rate, conception rate, and body and placental weights, crown-rump length and sex ratio of the live fetuses. External, visceral and skeletal examinations on the live fetuses revealed no compound-related abnormalities. From the present results, it is considered that the no-effect doses of THR-221 for the parental mice and the fetuses are 1000 and 3000 mg/kg/day, respectively.

[Teratological study of cefodizime sodium in mice--intravenous administration during period of organogenesis].

Cefodizime sodium (THR-221) was intravenously administered at dose levels of 300, 1000 and 3000 mg/kg/day to pregnant mice on days 6-15 of gestation covering the period of organogenesis, and its effects on the dams (F0) and the fetal and postnatal development and fertility of their offspring (F1) were examined. In each group including a control group, 21-24 of the 33-38 F0 dams were submitted to cesarean section on day 18 of gestation, and the remaining 12-14 animals were allowed to litter normally and nurse their offspring until day 21 of lactation. In the F0 dams, no compound effects were seen in general conditions, body weight, food consumption, numbers of corpora lutea and implantations or duration of gestation. The thymus weights of the cesarean sectioned dams at 1000 and 3000 mg/kg/day were less than the control values, but no difference appeared in weights of the other main organs between any treatment group and the control group. No gross visceral abnormalities were noted at any dose. In F1 near-term fetuses, the compound administration exerted no effects on their development at any dose, except the decreased body weight and crown-rump length noted at 3000 mg/kg/day. No difference occurred in incidence of fetuses with external, visceral or skeletal anomalies between any treatment group and the control group. Normally delivered F1 offspring in the treatment groups exhibited no changes from the controls in postnatal development, reflexes, 21-day survival index or behavioral test performance. Skeletal examination on F1 offspring which died during lactation and autopsies on the other animals at the end of study revealed no compound-related abnormalities. The fertility of the F1 mice was normal at all doses, and there were no compound effects on the F1 dams or their near-term fetuses or newborn pups. From the present results, it is considered that the no-effect doses of THR-221 for the maternal mice and their offspring are 300 and 1000 mg/kg/day, respectively.

[Effects of pentoxifylline on 5-hydroxytryptamine in the mouse brain].

Effects of combined administration of pentoxifylline (PTX) and precursors of 5-hydroxytryptamine (5-HT) on behavior and cerebral contents of 5-hydroxytryptophan (5-HTP), 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) were examined in mice. The intraperitoneal administration of PTX at 100 mg/kg to mice pretreated with pargyline (100 mg/kg, i.p.) significantly increased the frequency of head twitches induced by 5-HTP (25 mg/kg, i.p.), and the effect of PTX was approximately 2 times more potent than that of other methylxanthines. In these mice, the cerebral contents of 5-HTP, 5-HT and 5-HIAA were also elevated significantly. However, PTX itself had no effect to induce head twitch response or to increase the contents of the indoles in mice. When administered in combination with tryptophan (100 mg/kg, i.v.) to pargyline-pretreated mice, PTX (100 mg/kg, i.p.) did not affect the contents of the indoles in the brain. These results suggest that PTX may have an effect to promote transport of exogenous 5-HTP into the neurons, besides the 5-HT turnover-increasing effect common to methylxanthine derivatives.

[Pre-operative transcatheter arterial embolization for the patients with locally advanced breast cancers--a newly deviced thrombotic material].

Anticancer agents, Mitomycin C and Adriamycin, were immobilized on absorbable gelatin materials, respectively with a blood clotting factor, Factor XIII and thrombin with a special technique we have developed. The materials were clinically applied as thrombotic agent in pre-operative therapeutic transcatheter arterial embolization (TAE) to 5 patients with locally advanced breast cancer. About 1 week after TAE, the regression of the primary tumor was found in 4 out of 5 patients; all patients received currative operation without any trouble. The histological examinations of the excised specimens showed remarkable degenerative changes (IIA and IIB: Shimosato's classification) of cancer cells not only in the primary tumor, but also in the metastatic regional lymph node. No complications due to the TAE were observed. These findings strongly suggest that this newly deviced materials are quite effective in pre-operative treatment for the locally advanced breast cancers.

Renal arteriovenous malformations: transcatheter embolization and follow-up.

Transcatheter embolization was performed in three patients with renal arteriovenous malformations with multiple arterial feeders. The patients were followed angiographically for 6--14 months. In two cases, the malformation disappeared with focal loss of renal tissue in the region of embolization. In the third, there was recurrence of hematuria. Angiography demonstrated incomplete embolization, but the arteriovenous malformation had diminished.

Accessory left gastric artery arising from left hepatic artery: angiographic study.

An accessory left gastric artery may arise from the left hepatic artery and follow a course similar to an intrahepatic artery. In 98 hepatic angiograms an accessory left gastric artery was observed in 14 (14.2%). The site of origin of this artery was from the left hepatic artery proximal to the umbilical point. An esophageal branch was found in 10 cass (71.4%). The possibility of an accessory left gastric artery should be kept in mind during angiography of the left hepatic lobe and gastric fundus. A gastric wall stain in the capillary phase produced by this vessel may mimic a hepatic tumor.