Chymase inhibition attenuates monocrotaline-induced sinusoidal obstruction syndrome in hamsters.

Chymase stored in mast cells activates matrix metalloproteinase (MMP)-9, which may relate to the progression of sinusoidal obstruction syndrome (SOS). We investigated the preventive effect of a chymase inhibitor, TY-51469, on monocrotaline-induced SOS in hamsters. Hamsters were orally administrated with a single dose of monocrotaline (120 mg/kg) to induce SOS. Treatment with TY-51469 (1 mg/kg per day) or placebo had started 3 days before the monocrotaline administration. Two days after the monocrotaline administration, significant increases in aspartate aminotransferase, alanine aminotransferase and total bilirubin and a significant reduction of albumin were observed in plasma, but their changes were significantly attenuated by treatment with TY-51469. The numerous hepatic necrosis areas were observed in the placebo-treated group, but the ratio of necrotic area to total area in liver had been significantly reduced by treatment with TY-51469. Both chymase activity and MMP-9 level in liver were significantly augmented in the placebo-treated group. Furthermore, tumor necrosis factor (TNF)-α level in liver was also augmented in the placebo-treated group. However, the chymase activity and levels of MMP-9 and TNF-α were significantly attenuated in the TY-51469-treated group. Until 14 days after monocrotaline administration, survival rates in the placebo- and TY-51469-treated groups were 25% and 70%, respectively, and a significant difference was observed. In conclusion, chymase inhibition by TY-51469 may prevent the accelerating of severity in monocrotaline-induced SOS in hamsters.

Behavioural rhythm splitting in the CS mouse is related to clock gene expression outside the suprachiasmatic nucleus.

CS mice exhibit a spontaneous splitting in the circadian rhythm of locomotor activity under constant darkness, suggesting that they contain two weakly coupled oscillators in the circadian clock system regulating locomotor activity rhythm. In order to clarify whether the two oscillators are located in the suprachiasmatic nucleus (SCN), a site of the master circadian pacemaker in mammals, circadian rhythms in mRNA of mouse Period genes (mPer1, mPer2 and mPer3) in the SCN and cerebral cortex were examined during rhythm splitting by in situ hybridization. In the SCN, mPer1 and mPer2 showed a circadian rhythm with a single peak in both split and unsplit mice. The rhythms of mPer1 and mPer2 were slightly phase delayed during rhythm splitting in reference to the activity onset, but the phase relationship between the two rhythms was not changed. In the cerebral cortex, the expression of mPer1 and mPer2 underwent the bimodal fluctuation with peaks temporally corresponding to split activity components. The unsplit mice showed the circadian rhythms with a single peak. There was no difference in the mPer3 rhythms in either the SCN or the cerebral cortex between the split and unsplit mice. These results indicate that the circadian oscillations of mPer1, mPer2 and mPer3 in the SCN are not related to the rhythm splitting of CS mice. The split rhythms of the CS mice are suggested to be caused by uncoupling of oscillators located outside the SCN from the SCN circadian pacemaker.

Circadian activity rhythm in methamphetamine-treated Clock mutant mice.

It is well established that the Clock gene is essential for expressing circadian activity rhythms in mammals under constant darkness. The Clock gene product is a positive component of a molecular feedback loop which is assumed to generate the circadian rhythm. On the other hand, chronic treatment of methamphetamine (MAP) induces locomotor activity rhythm in a circadian domain, which is independent of the suprachiasmatic nucleus (SCN) and is driven by a pacemaker outside the SCN. However, it is not known whether the pacemaker outside the SCN possesses a similar molecular mechanism to that in the SCN. Here we show that MAP restores locomotor activity rhythm in arrhythmic homozygous Clock mutant (Clock/Clock) mice under constant darkness. This result indicates that the Clock mutation does not affect the MAP-induced locomotor rhythm.

Phase-advance shifts of human circadian pacemaker are accelerated by daytime physical exercise.

Effects of forced sleep-wake schedules with and without physical exercise were examined on the human circadian pacemaker under dim light conditions. Subjects spent 15 days in an isolation facility separately without knowing the time of day and followed a forced sleep-wake schedule of a 23 h 40-min period for 12 cycles, and physical exercise was imposed twice per waking period for 2 h each with bicycle- or rowing-type ergometers. As a result, plasma melatonin rhythm was significantly phase advanced with physical exercise, whereas it was not changed without exercise. The difference in phase was already significant 6 days after the start of exercise. The amplitude of melatonin rhythm was not affected. A single pulse of physical exercise in the afternoon or at midnight significantly phase delayed the melatonin rhythms when compared with the prepulse phase, but the amount of phase shift was not different from that observed in the sedentary controls. These findings indicate that physical exercise accelerates phase-advance shifts of the human circadian pacemaker associated with the forced sleep-wake schedule.

Circadian pattern, light responsiveness and localization of rPer1 and rPer2 gene expression in the rat retina.

Circadian expression, light-responsiveness and localization of clock genes, rPer1 and rPer2, were examined in the rat retina under constant darkness. A significant circadian variation was detected in rPer2 transcript levels with a peak at ZT14, but not in the rPer1. A light pulse given after constant darkness of 3 days increased both rPer1 and rPer2 expression phase-dependently, while rPer1 was induced at more times than rPer2. A major site of these gene expression within the retina was the inner nuclear layer. These findings indicate that rPer1 and rPer2 genes play different roles in the generation and regulation of circadian rhythms in the retina from those in the suprachiasmatic nucleus.

Clock gene expressions in the suprachiasmatic nucleus and other areas of the brain during rhythm splitting in CS mice.

The CS mouse is a mutant strain which displays spontaneous splitting in the circadian locomotor rhythm under continuous darkness. To clarify whether the rhythm splitting occurs in the suprachiasmatic nucleus (SCN) where the mammalian circadian clock is located, the circadian rhythmicities of mammalian clock genes, mPer1, mBMAL1 and mClock, were examined in the SCN and cerebral cortex during rhythm splitting. The circadian profiles of the clock genes during rhythm splitting were essentially the same as those observed under unsplit conditions. However, the mPer1 gene expression throughout the day was bimodal in the piriform and cingulate cortices, peaking in correspondence with two split components of behavioral rhythm. These results indicate that the circadian profiles of three clock gene expressions in the SCN are not consistent with the overt circadian locomotor rhythm, suggesting that the site of rhythm splitting is somewhere outside the SCN, or alternatively different subregions or other clock genes in the SCN are involved in rhythm splitting.

Clock genes outside the suprachiasmatic nucleus involved in manifestation of locomotor activity rhythm in rats.

Chronic treatment of methamphetamine (MAP) in rats desynchronized the locomotor activity rhythm from the light-dark cycle. When the activity rhythm was completely phase-reversed with respect to a light dark-cycle, 24 h profiles were examined for the clock gene (rPer1, rPer2, rBMAL1, rClock) expressions in several brain structures by in situ hybridization, and for the pineal as well as plasma melatonin levels. In the MAP-treated rats, the rPer1 expression in the suprachiasmatic nucleus (SCN) showed a robust circadian rhythm which was essentially identical to that in the control rats. Circadian rhythms in pineal as well as plasma melatonin were not changed significantly in the MAP-treated rats. However, robust circadian rhythms in the rPer1, rPer2 and rBMAL1 expressions detected in the caudate-putamen (CPU) and parietal cortex were completely phase-reversed in the MAP-treated rats, compared with those in the control rats, indicating desynchronization from the SCN rhythm. Such desynchronization was not observed in the circadian rhythms of clock gene expression in the nucleus accumbens and cingulate cortex. The circadian rClock expression rhythm in the MAP-treated rats was not phase-reversed in the CPU and parietal cortex. These findings indicate that the locomotor activity rhythm in rats is directly driven by the pacemaker outside the SCN, in which rPer1, rPer2 and rBMAL1 in the CPU and parietal cortex are involved.

Amplitude reduction of plasma melatonin rhythm in association with an internal desynchronization in a subject with non-24-hour sleep-wake syndrome.

The plasma melatonin rhythm was measured longitudinally in a subject with non-24-h sleep-wake syndrome, and the amplitude and area under the curve (AUC) of the melatonin rhythm were investigated in relation to the sleep-wake cycle. When the melatonin rhythm and sleep-wake rhythm were internally desynchronized, the amplitude and the AUC were reduced significantly. These parameters were not influenced by external melatonin administration. These results suggest that a causal relationship between the reduction of circadian oscillation and internal desynchronization exists in this subject.

The significance of microsatellite instability in predicting the development of metachronous multiple colorectal carcinomas in patients with nonfamilial colorectal carcinoma.

BACKGROUND: Patients with metachronous multiple colorectal carcinomas have been reported to have a higher frequency of a family history of colorectal carcinoma, associated colorectal adenomas, and extracolonic malignancies. These clinicopathologic factors also are considered to be related to the development of metachronous multiple colorectal carcinomas after surgery for colorectal carcinoma. In this article, the authors investigated whether genetic markers such as microsatellite instability (MSI) were helpful in predicting the development of metachronous multiple colorectal carcinomas. METHODS: Between 1990-1997, 312 colorectal carcinoma patients underwent yearly surveillance colonoscopy after surgery. Among these patients, there were 19 with nonfamilial colorectal carcinoma in whom metachronous multiple colorectal carcinomas were diagnosed during the yearly surveillance colonoscopy. A control group was comprised of 28 patients who did not demonstrate either synchronous or metachronous carcinomas over a period of > or =5 years. Six microsatellite markers (D2S123, D3S1029, D3S1611, TP53, Mfd26, and Mfd36) were used to determine MSI by polymerase chain reaction. RESULTS: The frequency of MSI positive cases was significantly higher in patients with sporadic metachronous multiple colorectal carcinomas than in those with a single carcinoma (17/19 [89%] vs. 4/28 [14%]; P<0.0001). In tumors occurring in the distal colon and rectum, the percentage of MSI positive carcinomas was significantly higher in the patients with metachronous multiple carcinomas than in those with a single carcinoma (13/15 [87%] vs. 0/19 [0%]; P<0.0001). No such difference was observed in the proximal colon. CONCLUSIONS: Based on the findings of the current study, the analysis of MSI in sporadic carcinomas of the distal colon and rectum may be helpful in predicting the development of metachronous multiple colorectal carcinomas.

Intestinal absorption of acyclovir beta-glucoside: comparative study with acyclovir, guanosine, and kinetin beta-glucoside.

PURPOSE: To characterize the intestinal absorption of a beta-glucose conjugate of acyclovir (9-[(2-hydroxyethoxy) methyl] guanine, ACV) and compare it to ACV and its analogues in terms of stability and transport by Na+/glucose cotransporter (SGLTI). METHODS: ACVbeta(glc) was enzymatically synthesized using cellulase. Intestinal absorption experiments were performed with rat everted small intestine. Conformation of the glucose moiety was analyzed by NMR spectroscopy. RESULTS: The ACVbeta(glc) was stable on the mucosal side, and was transported to the serosal side in all regions of the small intestine. However, significant contribution of SGLTI to the transport of ACVbetaglc was not observed. NMR spectroscopic analysis indicated that the glucose conformation of ACVbeta(glc) was the 4C1 chair form, identical to beta-glucose or SGLT1-transportable beta-glucosides reported previously. Therefore, other factors such as molecular size and charge due to aglycone may cause no transport of ACVbeta(glc) by SGLT1. On the other hand, the hydrophilicity of ACVbeta(glc) was much higher than of ACV, suggesting water solubility-derived improvement of intestinal absorption of ACV. CONCLUSIONS: ACVbeta(glc) is stable and absorbable, but it is not transported by SGLT1. No involvement of SGLT1 in the ACVbeta(glc) transport is not due to glucose conformation.

Important microsatellite markers in the investigation of replication errors (RER) in colorectal carcinomas.

BACKGROUND: DNA replication errors (RER) have been found in hereditary nonpolyposis colorectal carcinomas and in sporadic colorectal carcinomas. The incidence of RER depends on which and how many markers are examined. The main purpose of the present study was to determine the key markers for detecting RER most efficiently. METHODS: The RER status of 76 sporadic advanced colorectal carcinomas in the proximal colon were investigated. Seven microsatellite markers (D2S123, D3S1029, D3S1611, D2S72, TP53, Mfd26 and BAT26) were chosen to determine the RER status by PCR using the non-Rl method, because these seven markers have frequently been used in other studies and also detect RER. RESULTS: It was found that 44.7% of sporadic colorectal advanced carcinomas in the proximal colon (34 of 76) showed RER at one or more loci. Among these 34 cases, RER was present at three or more markers (severe RER) in 22. All 22 of these cases showed RER at BAT26 and TP53. The other 12 cases with RER showed RER at one or two markers (mild RER). Eleven of these 12 cases (91%) showed RER at Mfd26 and there were one or two cases with mild RER at each of the other loci. CONCLUSIONS: When one intends to analyze routinely a large number of cases, an analysis of two or three markers (Mfd26 and BAT26 or TP53) is considered to be sufficient for detecting mild and severe RER.

Intestinal absorption of stable cyclic dipeptides by the oligopeptide transporter in rat.

Intestinal absorption of four cyclic dipeptides was studied in the everted small intestine of the rat. Cyclic seryltyrosine (cyclo(Ser-Tyr)) was stable enough to be transported whereas linear seryltyrosine was not. The absorption clearance of cyclo(Ser-Tyr) was concentration-dependent, and for cyclo(Ser-Tyr) at 125 microM decreased in the presence of glycylsarcosine (10 mM) or cephalexin (10 mM), which were reported to be absorbed by oligopeptide transporter. The absorption clearance was also reduced at 4 degrees C and in the presence of 1 mM dinitrophenol. Kinetic analysis of cyclo(Ser-Tyr) absorption showed that Km and Vmax were 19.8 microM and 0.295 nmol min(-1) cm(-1), respectively. It was also suggested that cyclic aspartylphenylalanine and cyclic histidylphenylalanine were absorbed by oligopeptide transporters, but cyclic histidylproline was not. The absorption clearance of cyclo(Ser-Tyr) in the control was much higher than the value of the correlation line representing a plot of passive transport (which was obtained from the absorption clearance of cyclic peptides in the presence of glycylsarcosine (10 mM)) against hydrophobicity (oil-water partition coefficient). These results indicate that cyclo(Ser-Tyr) is absorbed by the oligopeptide transporter.

Intestinal absorption of stable cyclic glycylphenylalanine: comparison with the linear form.

The absorption, especially the stability and transportability, of the cyclic peptide cyclic glycylphenylalanine (cyclo(Gly-Phe)) and the linear peptides glycylphenylalanine, glycyl-D-phenylalanine and phenylalanylglycine have been studied in rat small intestine. Linear peptides were degraded on the mucosal side and only glycyl-D-phenylalanine appeared on the serosal side. However, cyclo(Gly-Phe) was stable on the mucosal side and appeared on the serosal side. Furthermore, the absorption clearance of cyclo(Gly-Phe) was higher than that of glycyl-D-phenylalanine. In the presence of the peptidase inhibitor bestatin, the degradation of linear peptides was reduced and linear peptides appeared on the serosal side, but only phenylalanylglycine, which is transported by the oligopeptide transporter, was absorbed faster than cyclo(Gly-Phe). The absorption clearance of cyclo(Gly-Phe) was reduced as its concentration was increased from 125 microM to 500 microM. Furthermore, the absorption clearance of cyclo(Gly-Phe) at 125 microM was reduced at 4 degrees C or in the presence of glycylsarcosine and cephalexin, which are transported by the oligopeptide transporter. These results indicated that cyclo(Gly-Phe) was stable enough to be absorbed and was transported in part by the oligopeptide transporter rather than completely by passive diffusion.

Epidemiologic study of carcinoma in situ of the cervix.

From 1950 to 1979, 1,248 cases of cancer in situ were treated in the Department of Gynecology, Cancer Institute Hospital, Tokyo. Detailed data were obtained from 585 of the patients by interview. Many patients had had their first sexual experience at an early age. Many had had more than two sexual partners, and there was a large discrepancy between the age at first sexual intercourse and at first marriage. The above factors seem to be related to the development of cancer in situ.

[Preoperative diagnosis for stage II endometrial carcinoma using endocervical conization].

It is generally difficult to identify stage II endometrial carcinoma correctly. To differentiate cervical involvement, endocervical conization was performed on 31 patients with endometrial cancer between May 1982 and September 1983. The preoperative diagnosis of stage II endometrial carcinoma was confirmed by postoperative histologic examination. In stage I patients, there was no microscopic cervical involvement after the operation. Fractional curettage and hysteroscopy are commonly used to detect stage II endometrial carcinoma. However, these examinations may overlook cervical stromal invasion. Endocervical conization is a simple procedure and highly useful to detect cervical stromal involvement. We recommend the of endocervical conization combined with hysteroscopy to diagnose stage II endometrial carcinoma.

[Therapeutic results of malignant ovarian tumor].

One hundred and fifty patients with malignant ovarian tumors were treated in the Cancer Institute Hospital between 1949 and 1977. Most (103; 68.7%) of the patients had simple primary ovarian cancer. A retrospective study was performed in these 103 patients to investigate the relationship between the 5-year survival rate and the clinical stage and primary treatment. The 103 patients were classified according to FIGO criteria. The survival rates for stages I, II, III and IV were 73.7%, 50.5%, 17.4% and 0%, respectively. As the primary treatment, operations were performed on 92 of the 103 patients. To conclude, of primary importance in the treatment of ovarian cancer is the surgical, complete as possible removal of the tumors. Postoperatively, radiotherapy and chemotherapy should be used. Recently, second look operations have come into wider use. General procedures for the surgical and postoperative treatment of progressive ovarian cancer have been established. Although surgical procedures for the treatment of early ovarian cancer have also been established, there are no consistent procedures for postoperative treatment. Since the incidence of ovarian cancer is increasing, the development of appropriate methods for early diagnosis and treatment of this carcinoma is eagerly awaited.

Psammoma bodies found in cervicovaginal and/or endometrial smears.

Eight cases of the unusual occurrence of psammoma bodies associated with adenocarcinoma cells in cervicovaginal and/or endometrial smears are presented. Three of the eight cases were advanced ovarian cancers, four were primary endometrial adenocarcinomas, and one was a metastasis of breast cancer to the pouch of Douglas. From our histologic and histochemical observations, psammoma bodies seem to be secretory rather than degenerative in origin.