Effect of oxaliplatin-based chemotherapy on chemosensitivity in patients with peritoneal metastasis from colorectal cancer treated with cytoreductive surgery and hyperthermic intraperitoneal chemotherapy: proof-of-concept study.

BACKGROUND: Chemosensitivity testing, including collagen gel droplet-embedded culture drug sensitivity test, has proven to be a useful tool in therapeutic decision-making. This retrospective analysis investigated chemosensitivity testing of peritoneal metastases collected during cytoreductive surgery (CRS), and its impact on survival in patients with colorectal cancer. METHODS: All patients with peritoneal metastasis from colorectal cancer who underwent CRS with or without hyperthermic intraperitoneal chemotherapy (HIPEC) between November 2008 and October 2014 were included. The growth inhibition rate was expressed as the ratio between the image density after treatment (T) and that before treatment (control, C). Tumours with a reduction in T/C ratio of less than 20 per cent were defined as resistant and those with a reduction of 20 per cent or more as sensitive. Groups were compared for overall (OS) and disease-free (DFS) survival. RESULTS: Of 84 eligible patients, 81 received neoadjuvant chemotherapy (NACT), including 56 patients with an oxaliplatin-based regimen. Mean(s.d.) follow-up was 23·4(22·9) months. The median overall survival of all patients was 19·0 (i.q.r. 5·7-36·1) months, with a progression-free survival time of 10·1 (4·5-17·0) months. Patients who received oxaliplatin-based NACT had significantly altered chemosensitivity to oxaliplatin; only 20 of 51 such patients showed chemosensitivity to oxaliplatin compared with 16 of 24 who did not undergo oxaliplatin-based NACT (P = 0·046). However, patients who showed chemoresistance to oxaliplatin had similar OS to those with chemosensitivity (18·8 versus 18·1 months; P = 0·835). The choice of HIPEC agents in patients who received oxaliplatin-based NACT did not significantly influence survival (oxaliplatin versus mitomycin C: median OS 20·6 (10·9-24·8) versus 19·0 (10·5-34·6) months, P = 0·811; DFS 6·6 (2·8-25·7) versus 9·3 (4·1-13·9) months, P = 0·191). CONCLUSION: Patients who had oxaliplatin-based NACT showed a higher rate of chemoresistance to oxaliplatin at the time of CRS and HIPEC. The impact of chemosensitivity testing on OS remains unclear and needs further investigation.

Disruption of ATM in p53-null cells causes multiple functional abnormalities in cellular response to ionizing radiation.

ATM is a member of the large phosphatidylinositol-3 kinase family and plays an important role in cellular response to DNA damage. To further define the physiological roles of ATM at the cellular level, we created an isogenic set of stable cell lines differing only in their ATM status from the chicken B cell line DT40 by targeted integration. These stable DT40 cell lines, as most of transformed chicken cell lines, do not express p53. However, ATM-/- DT40 cells displayed retarded cellular proliferation, defective G2/M checkpoint control and radio-resistant DNA synthesis. Furthermore, ATM-/- DT40 cells were sensitive to ionizing radiation and showed highly elevated frequencies of both spontaneous and radiation-induced chromosomal aberrations. In addition, a slight but significant reduction in targeted integration frequency was observed in ATM-/- DT40 cells. These results suggest that ATM has multiple p53-independent functions in cell cycle checkpoint control and in maintenance of chromosomal DNA. These ATM deficient DT40 clones therefore provide a useful model system for analysing p53-independent ATM functions.

Protective roles for ATM in cellular response to oxidative stress.

ATM (ataxia telangiectasia mutated), the gene mutated in ataxia telangiectasia, is related to a family of large phosphatidylinositol 3-kinase domain-containing proteins involved in cell cycle control and DNA repair. We found that ATM(-/-) DT40 cells were more susceptible than wild-type cells to apoptosis induced not only by ionizing radiation and bleomycin but also by non-DNA-damaging apoptotic stimuli such as C(2)-ceramide. Furthermore, the apoptosis induced by C(2)-ceramide and H(2)O(2) was blocked by anti-oxidants, indicating that the ATM(-/-) DT40 cells had a heightened susceptibility to apoptosis induced by reactive oxygen intermediates (ROI), presumably due to defective ROI-detoxification activities. In support of this hypothesis, we found that more ROI were generated in ATM(-/-) DT40 cells than in wild-type cells, following treatment with the above apoptotic stimuli. These results indicate that ATM plays important roles in the maintenance of the cell homeostasis in response to oxidative damage.

Altering the timing of human chorionic gonadotropin injection according to serum progesterone (P) concentrations improves embryo quality in cycles with subtle P rise.

OBJECTIVE: To investigate the effect of earlier administration of hCG according to serum concentrations of P on the outcome of IVF-ET in cycles in which a subtle rise in serum P (1.0 to 2.0 ng/mL; conversion factor to SI unit, 3.180) occurred during the follicular phase. DESIGN: Retrospective study. PATIENTS: A total of 110 infertile women underwent 124 cycles of IVF-ET at Tottori University Hospital. MAIN OUTCOME MEASURES: Serum was obtained daily or every 12 hours from day 7 until the administration of hCG. Serum E2 and P concentrations were measured by RIA. In 19 of 36 subtle P rise cycles, hCG injection was given when the levels of serum P exceeded 1.0 ng/mL ("rescued" subtle P rise). Parameters of IVF outcomes for the no P rise, the subtle P rise, and the rescued subtle P rise cycles were compared. RESULTS: The mean day of hCG administration in the rescued cycles was 1 day earlier than those of the subtle P rise and no P rise cycles. The mean number of oocytes collected was significantly higher in the subtle P rise and rescued P rise cycles than in the no P rise cycles. The mean follicular diameter on the day of hCG administration was 13.9 mm in the rescued cycles, significantly smaller than those of the no P rise and subtle P rise cycles. However, there was no significant difference in the cleavage rates between the three groups. The rate of embryonic development beyond four-cell stage was increased significantly in the rescued cycles and no P rise cycles versus the subtle P rise cycles. Embryos obtained in the no P rise and rescued cycles were of better morphological quality than those obtained in the P rise cycles. The implantation rate was significantly higher in the rescued cycles than in the P rise cycles. CONCLUSION: The data suggest that, if hCG is administered when a subtle P rise is detected, embryo quality and subsequent implantation rate can be improved.

Hydrophobicity parameter of diazines. II: Analysis and prediction of partition coefficients of disubstituted pyrazines.

The partition coefficient in a 1-octanol-water system (log P) of a number of disubstituted pyrazines was measured. The increment produced by introduction of two substituents [pi(disubst)PR] was analyzed in terms of the pi value of substituents in monosubstituted pyrazine (pi 2PR) and other physicochemical substituent parameters, considering electronic and steric interactions operating bidirectionally between individual partners of substituents and aza-functions. When the substituents were nonamphiprotic and their steric interaction was not significant, the pi (disubst)PR value was approximately predicted by the summation of the pi 2PR values of corresponding substituents. For amphiprotic substituents, a correction for electronic interactions between the two substituents was needed but it was less important than that required in disubstituted benzenes. The results suggest that the difference in the pi values between disubstituted pyrazine and benzene systems was mostly governed by that between monosubstituted pyrazine and benzene systems with component substituents. In 2,6- and 2,3-disubstituted derivatives, bulky substituents had steric effects in such a way that the log P value was lowered.

Hydrophobicity parameter of diazines. III: Relationship of partition coefficients of monosubstituted diazines and pyridines in different partitioning systems.

The logarithm of the 1-octanol-water partition coefficient value (log Poct) was compared with those from CHCl3-water (log PCL) and di-n-butyl ether-water (log PE) for (di)azines substituted singly by nonhydrogen-bonding and hydrogen-accepting substituents (2-substituted pyrazines, 2-substituted pyrimidines, 5-substituted pyrimidines, and 2-substituted pyridines). The difference between log Poct and log PCL for diazines was primarily governed by the number of hydrogen-bonding sites in the substituent. For 2-substituted pyridines, the difference in the hydrogen-bonding association of the ring N-atom with octanol from that with CHCl3 was also significant. In the relationship between log Poct and log PE, the hydrogen-bonding solvations of the ring N-atom(s), as well as the hydrogen-accepting substituent with octanol, should be taken into account because the butyl ether acts as a nonhydrogen-bonding solvent.

TRH stimulation test in healthy elderly: paradoxical response of growth hormone is abnormal in normal aging.

The purpose of this study was to measure TSH (thyrotropin), PRL (prolactin) and GH (growth hormone) response to a TRH (thyrotropin releasing hormone) stimulation test in 37 healthy elderly male subjects and determine if these responses correlate with the incidence of mental decline; and also, to determine if these responses occur more frequently in the elderly than in the general population. A hyperresponse of TSH was documented in 7 of our 37 elderly male subjects, a delayed response of TSH in 4, a low response of PRL in 29 and a significant paradoxical response of GH in 2. Those subjects with a significant paradoxical response of GH were 82 and 83 years of age and their HDS was 20.0 and 25.5, respectively. We did not find a causal relationship between mental decline and the paradoxical response of GH, but our data suggests that the paradoxical response of GH is not normal in elderly older than 80 years old.

[Seminal vesicle cyst with ipsilateral renal agenesis: a case report].

A 26-year-old man was referred to our hospital for further evaluation of a cystic lesion in the pelvis. He was completely asymptomatic, although he experienced gross hematuria 10 months before his visit. Drip infusion pyelography (DIP) and abdominal computerized tomography (CT) showed a cystic structure behind the bladder and absence of the left kidney. Transrectal ultrasonography (TRUS) suggested left ejaculatory duct obstruction and seminal vesicle deformity. On cystoscopy a large bulging mass under the left side of the bladder was seen. The left half of the trigone and left ureteral orifice were absent. Vasovesiculography demonstrated dilated and tortuous left seminal vesicle. Three days after this procedure, the patient suffered left epididymitis. It did not respond rapidly to antibiotic therapy, so percutaneous drainage of the cyst was performed. He has been completely free of symptoms and no seminal vesicle cysts have been detected on TRUS at five months follow-up.

Surgical treatment for peritoneal carcinomatosis from gastric cancer.

This review describes the latest surgical treatments for peritoneal carcinomatosis (PC) arising from gastric cancer. Systemic chemotherapy is less effective against PC because of the existence of the blood-peritoneal barrier. Accordingly, perioperative intraperitoneal chemotherapy plus cytoreductive surgery (CRS) is a new trend of multidisciplinary therapy for PC. Intraperitoneally administered drugs penetrate directly into the peritoneal dissemination, resulting in the high loco-regional intensity of drugs. A new bidirectional chemotherapy called neoadjuvant intraperitoneal/systemic chemotherapy (NIPS) has been developed. After NIPS, the disappearance of PFCCs has been reported, and the incidence of complete cytoreduction has increased accordingly. Complete cytoreduction, a low peritoneal carcinomatosis index, and negative PFCCs are significant favorable prognostic factors. Hyperthermic intraperitoneal chemotherapy (HIPEC) after CRS is associated with improved survival with an acceptable postoperative mortality and morbidity. Early postoperative intraperitoneal chemotherapy (EPIC) has also contributed to improving survival after CRS.

[Studies on partial zona dissection in in vitro fertilization for severe male infertility].

The effects of partial zona dissection (PZD) were investigated in mouse in vitro fertilization (IVF) with an oligospermic model and then PZD was applied to patients with severe male infertility. Micromanipulated (PZD-treated) and zona-intact (control) mouse oocytes were inseminated with 1 x 10(4) (low sperm concentration) and 1 x 10(5), 1 x 10(6) (normal sperm concentration) motile sperm/ml. In mouse IVF with 1 x 10(4) sperm/ml, the fertilization rate in those PZD-treated were significantly higher than that in the control (21.3% vs 8.1%, p < 0.01). No polyspermy was observed. No significant difference in the rate of development to the blastocyst stage was seen between the PZD-treated and the control (71.0% vs 65.0%). The incorporation rate of 3H-leucine in embryos at the 2-cell and blastocyst stages also did not differ between the two groups of mice. Sixteen patients with severe male infertility who had failed to fertilize for the previous three consecutive IVF cycles were enrolled in the PZD protocol. The fertilization rate in PZD-treated cycles was significantly improved (19.6% in PZD-treated vs 0% in control cycles, p < 0.001). In ten patients who underwent replacement with PZD-treated embryos, one biochemical pregnancy occurred. The present study suggests that although PZD is useful for severe male infertility, further investigation is needed to evaluate PZD in assisted fertilization.

Transformations of (+-)-salsolinol into optically active O- and/or N-methylated derivatives by several Papaveraceae plants and their tissue-cultured cells.

(+-)-Salsolinol [3], a substance possibly inducing parkinsonism, was biotransformed into optically active O- and/or N-methylated salsolinols 5, 6, and 7 by several Papaveraceae plants and tissue-culture cells derived from these plants. The bioconversion of racemic salsolinol into optically active tetrahydroisoquinolines has never been shown to occur, either in animals or in plants.

Reinvestigation of the conformations of a variety of hexahydrobenzo[c]phenanthridine alkaloids by 470 MHz PMR and 50 MHz CMR spectroscopy.

The high resolution pmr and cmr spectra of a variety of benzo[c]phenanthridine alkaloids and their CF3COOD salts were examined. The chemical shift of H-14 was found to be a reliable indicator of the orientation of the N-methyl group. The conformations of the C rings were assigned on the basis of the coupling constants between H-11 and the two H-12 protons. On the basis of the pmr spectra of (+)-14-epicorynoline (9) and (+/-)-14-epicorynoline-6,6,12 alpha-d3 (13), a revision of certain previous C ring conformations is indicated.

c-Abl tyrosine kinase is not essential for ataxia telangiectasia mutated functions in chromosomal maintenance.

c-Abl is activated by DNA damage in an ataxia telangiectasia mutated (ATM)-dependent manner and plays important roles in growth arrest and apoptosis induced by DNA damage. c-Abl also interacts physically and functionally with Rad51, a key molecule in homologous recombinational (HR) DNA repair. To study further the roles of c-Abl in HR DNA repair, we generated c-Abl(-/-) and ATM(-/-)/c-Abl(-/-) mutant cell lines from a chicken B lymphocyte DT40 cell line, comparing the phenotypes of these mutants to those of ATM(-/-) DT40 cells that we had created previously. We found that the time course of radiation-induced Rad51 focus formation is abnormal in ATM(-/-) DT40 cells, consistent with the observation that ATM(-/-) DT40 cells display hypersensitivity to ionizing radiation and highly elevated frequencies of both spontaneous and radiation-induced chromosomal aberrations. In contrast, c-Abl(-/-) cells did not show these ATM-related defects in their cellular response to radiation, nor did the disruption of c-Abl in ATM(-/-) DT40 cells exacerbate these ATM-related defects. However, c-Abl(-/-) DT40 cells, but not ATM(-/-) DT40 cells, were resistant to radiation-induced apoptosis, indicating an important role for c-Abl in this cellular response to ionizing radiation. These results therefore indicate that, although ATM plays an important role in genome maintenance, c-Abl is not essential for this ATM function. These findings suggest that c-Abl and ATM play important roles in the maintenance of the cell homeostasis in response to DNA damage that are, at least in part, independent.

The controlling role of ATM in homologous recombinational repair of DNA damage.

The human genetic disorder ataxia telangiectasia (A-T), caused by mutation in the ATM gene, is characterized by chromosomal instability, radiosensitivity and defective cell cycle checkpoint activation. DNA double-strand breaks (dsbs) persist in A-T cells after irradiation, but the underlying defect is unclear. To investigate ATM's interactions with dsb repair pathways, we disrupted ATM along with other genes involved in the principal, complementary dsb repair pathways of homologous recombination (HR) or non-homologous end-joining (NHEJ) in chicken DT40 cells. ATM(-/-) cells show altered kinetics of radiation-induced Rad51 and Rad54 focus formation. Ku70-deficient (NHEJ(-)) ATM(-/-) chicken DT40 cells show radiosensitivity and high radiation-induced chromosomal aberration frequencies, while Rad54-defective (HR(-)) ATM(-/-) cells show only slightly elevated aberration levels after irradiation, placing ATM and HR on the same pathway. These results reveal that ATM defects impair HR-mediated dsb repair and may link cell cycle checkpoints to HR activation.

Age-dependent decline in bone nodule formation stimulating activity in rat serum is mainly due to the change in the corticosterone level.

The replacement of fetal bovine serum with rat serum in a culture medium brought about a marked increase in the formation of mineralized bone nodules (BN) in primary cultures of rat calvarial cells. These effects of rat serum were most prominent when added during the early phase of the culture, indicating that the serum factor mainly acts on the cells during the growing phase. A significant increase in BN formation was observable at final rat serum concentration as low as 1%, and the effect was dependent on serum concentration, at least up to 10%. The addition of rat serum also increased alkaline phosphatase (ALP) activity, collagen synthesis, and DNA synthesis in calvarial cells. BN formation stimulating activity was extractable with ethyl acetate. The ethyl acetate extract was purified by TSK-GEL OH-120 column chromatography by monitoring the stimulation of ALP activity in ROS 17/2.8 cells. The chromatographic behavior of the ALP activity was found to be identical to that of corticosterone, the major glucocorticoid in rodents and the preincubation of the purified fraction with anticorticosterone antibody abolished the ALP stimulating activity. These results suggest that BN formation stimulating activity in rat serum is mainly attributable to corticosterone. The concentration of serum corticosterone decreased with age in parallel with BN formation stimulating activity, which suggests that the physiological level of corticosterone may have a regulatory role in the maintenance of osteoblast function.

Reduced implantation rate associated with a subtle rise in serum progesterone concentration during the follicular phase of cycles stimulated with a combination of a gonadotrophin-releasing hormone agonist and gonadotrophin.

Our objective was to assess the effects of subtle increases in serum progesterone concentration (1.0-2.0 ng/ml) on the outcome of in-vitro fertilization (IVF), particularly on the quality of embryos, during the follicular phase of cycles stimulated with gonadotrophin-releasing hormone agonist (GnRHa) and human menopausal gonadotrophin (HMG). A total of 97 patients underwent 116 cycles of IVF and were stimulated with a combination of HMG and GnRHa. They were divided into two groups: those with a subtle progesterone rise and those with no progesterone rise. The two groups were compared with respect to serum oestradiol, progesterone, immunoreactive luteinizing hormone (I-LH), bioactive LH (B-LH), and results of IVF. The groups did not differ significantly in mean age or in total dose of HMG received. On the day that human chorionic gonadotrophin was administered, concentrations of oestradiol and progesterone were significantly higher in the subtle progesterone rise cycles than in the no progesterone rise cycles. In the no progesterone rise cycles, the percentages for embryos beyond the 4-cell stage, grade 1 embryos, and implantation rates were significantly higher than those in subtle progesterone rise cycles. The combination of GnRHa and HMG eliminated any significant rise in serum I-LH or B-LH concentration during the follicular phase, but did not suppress the subtle rise in progesterone. These results confirm our previous finding that a subtle progesterone rise adversely affects the outcome of IVF. It is also suggested that a reduction in embryo quality may influence the lower rate of implantation in subtle progesterone rise cycles.