Fixed dosing and pharmacokinetics of S-1 in Japanese cancer patients with large body surface areas.

BACKGROUND: S-1 is an oral anticancer agent that combines tegafur (FT) with 5-chloro-2,4-dihydroxypyridine (CDHP) and potassium oxonate. The recommended initial dose of S-1 is 120 mg/day for patients with a body surface area (BSA) of > or =1.5 m(2) in Japan. METHODS: We examined the effects of using this fixed dose on the pharmacokinetics of FT, CDHP, and active 5-fluorouracil (5-FU) on the basis of actual BSA. The pharmacokinetics was compared between patients with a BSA of 1.5-1.75 m(2) and those with a BSA of > or =1.75 m(2). RESULTS: The median areas under the time-concentration curves (AUCs) of 5-FU and CDHP were significantly lower in patients with a BSA of > or =1.75 m(2) than in those with a BSA of 1.5-1.75 m(2) (P = 0.005 and 0.006, respectively; Mann-Whitney U-test). There was no difference between the groups in the median AUC of FT. CONCLUSION: Systemic exposure to 5-FU is significantly lower in Japanese cancer patients with a large BSA of >1.75 m(2) who received the recommended fixed dose of S-1.

Pharmacokinetics of 5-fluorouracil in elderly Japanese patients with cancer treated with S-1 (a combination of tegafur and dihydropyrimidine dehydrogenase inhibitor 5-chloro-2,4-dihydroxypyridine).

S-1 is an oral anticancer agent that combines tegafur, a prodrug of 5-fluorouracil (5-FU), and 5-chloro-2,4-dihydroxypyridine (CDHP), an inhibitor of dihydropyrimidine dehydrogenase. We examined the effects of aging on the pharmacokinetics of the components of S-1. The median area under the concentration-time curve (AUC) of active 5-FU did not significantly differ between 10 patients 75 years or older and 53 patients younger than 75 years (P = 0.598, Mann-Whitney U test). It is interesting to note that the median oral clearance of tegafur in patients 75 years or older was significantly lower than that in patients younger than 75 years (P = 0.011). Furthermore, the median AUC of CDHP was significantly higher in patients 75 years or older than in those younger than 75 years (P = 0.004). This effect was caused by reduced renal function in the elderly, because CDHP is excreted in the urine by glomerular filtration. The opposing effects of aging on the oral clearance of tegafur and the AUC of CDHP may offset each other, leading to unchanged systemic exposure of 5-FU.

An ascidian homologue of vertebrate BMPs-5-8 is expressed in the midline of the anterior neuroectoderm and in the midline of the ventral epidermis of the embryo.

The ascidian tadpole larva is thought to be the prototype for the ancestral chordate. Although ascidians show a highly determinate mode of development, recent studies suggest significant roles of cell-cell interaction during embryogenesis. To elucidate the signaling molecules responsible for the cellular interaction, we investigated an ascidian homologue of the transforming growth factor beta (TGF-beta) superfamily. HrBMPa is an ascidian member of the 60A subclass of the BMP subfamily. Molecular phylogenetic analysis suggested that HrBMPa branched prior to further divergence of vertebrate BMPs-5-8. The zygotic expression of HrBMPa was initiated around gastrulation. HrBMPa transcripts were first evident in precursor cells of the spinal cord, notochord, epidermis and nervous system, although signals in the first two regions quickly disappeared. In neurulae and early tailbud embryos, transcripts were evident in the adhesive organ, midline of the anterior dorsal neuroectoderm and midline of both ventral and dorsal ectoderm, suggesting that HrBMPa plays a major role in neuroectodermal cell differentiation during embryogenesis. This HrBMPa expression profile resembled that of Xenopus BMP-7, implying a primordial function of BMP-7 among vertebrate BMPs-5-8.

Isolation and characterization of cDNA clones for beta-tubulin genes as a molecular marker for neural cell differentiation in the ascidian embryo.

The central nervous system (CNS) of an ascidian tadpole larva is composed of about 340 cells, the lineages of which are well documented. To elucidate the mechanisms underlying the neural induction of ascidians, appropriate molecular markers are required. In this study, to obtain an early differentiation marker of the neural cells, we isolated and characterized cDNA clones for two beta-tubulin genes (HrTBB1 and HrTBB2) of the ascidian Halocynthia roretzi. We found that the HrTBB1 and HrTBB2 amino acid sequences are highly conserved, with 91-98% identities to other invertebrate and vertebrate beta-tubulins. The expression of HrTBB1 was found to be maternal, while HrTBB2 is expressed both maternally and zygotically. We observed that the zygotic expression of HrTBB2 commences at the neural plate stage and is specific to cells of the differentiating CNS. In the larvae, HrTBB2 expression was restricted to cells of the CNS, some cells of the papilla and cells of the peripheral nervous system. These results indicate that HrTBB2 will be a useful early molecular marker for neural cell differentiation in the ascidian embryo.

Rare mutations of the growth suppressor genes involved in negative regulation of the cell cycle.

The growth suppressing activity of the retinoblastoma susceptibility gene product, pRb, is down regulated by cyclin-dependent kinases 4 and 6 (CDK4 and CDK6) whose activity is negatively regulated by CDK inhibitors of the p16 family. We have previously reported point mutations of the p16/CDKN2 gene in 4 (57%) of 7 oral squamous cell carcinoma (SCC) cell lines. In the current study, we examined the mutational status of CDK inhibitors, including 3 genes of the p16 family (p16, p15 and p18), in 50 human oral SCCs, and also additional results concerning their loss of heterozygosity in the regions of the p16, p15 and p18 genes. Our results demonstrated that 2 of 50 (4%) primary oral SCCs had nonsense mutations of the p16 gene, and 2 of 50 (4%) showed frameshift mutations of the p18 gene. However, we detected no mutation of the p15 gene in any of the 50 oral SCCs. In addition, no evidence of hypermethylation of the p16 gene was found in our series. To better understand the extent of alterations affecting chromosomes 9p21 (location of the p15/p16 genes) and 1p32 (location of the p18 gene), loss of heterozysity (LOH) on these locations was examined. LOH was detected in 16 of 34 (47%) informative samples that had no detectable mutation of the p15/p16 genes on 9p21, but we found no LOH at 1p32. These results strongly suggest that a putative tumor suppressor gene for oral SCC may be present on chromosome 9p21-22, while the p16, p15 and p18 genes play a minor role in the oncogenesis of this cancer.

Allelic loss on chromosome 22 in oral cancer: possibility of the existence of a tumor suppressor gene on 22q13.

In order to understand the detail of genetic alternation on chromosome 22, we performed polymerase chain reaction analysis of microsatellite polymorphisms corresponding to 13 loci on chromosome 22. We examined 33 primary carcinoma tissues, 5 metastatic tissues and corresponding normal tissues. We detected microsatellite instability (MI) in 14 (42.4%) of 33 cases in this study. Loss of heterozygosity (LOH) was observed in at least one locus in 24 (72. 7%) of the 33 cases. Among the loci examined, LOH was restricted to D22S274 on chromosome 22q13 in 11 (40.7%) of 27 informative cases. No significant correlation between histological differentiation and LOH was observed. These observations suggest that the incidence of LOH at chromosome 22q is high and is associated with the carcinogenesis of oral squamous cell carcinoma (SCC). The D22S274 locus may play an important role in the development of oral SCC and be the site harboring a putative tumor suppressor gene.

Enzyme immunoassay of pancreatic glucagon at the picogram level using beta-D-galactosidase as a label.

An enzyme immunoassay of pancreatic glucagon was established by using E. coli beta-D-galactosidease [EC 3.2.1.23] as a marker. In order to increase the sensitivity of the immunoassay, different peptides obtained from glucagon fragments were used to produce the enzyme conjugate and the immunogen. Antiserum N6E raised against C-terminal fragment peptide (15-29) could be diluted to more than 1 : 100,000 in the assay and was highly specific for pancreatic glucagon. The antiserum reacted well with the C-terminal fragment peptide (21-29) as well as another fragment peptide (15-29) and pancreatic glucagon. The enzyme immunoassay using antiserum N6E and fragment peptide (21-29)-enzyme conjugate could detect as little as 1 to 2 pg of glucagon. The mean recovery of glucagon added to serum specimens was 104% and the coefficients of variation were 3.7-14.5% (within assay) and 9.0-18.5% (between assay).

Standard lecithin/sphingomyelin and phosphatidylglycerol techniques compared with immunologic slide test.

A commercially available, rapid, technically simple, immunologic slide agglutination test for amniotic fluid (AF) phosphatidylglycerol has been compared with the lecithin/sphingomyelin (L:S) ratio and phosphatidylglycerol determined by thin-layer chromatography. The immunologic test lacks sensitivity when compared with phosphatidylglycerol measured by thin-layer chromatography, and both phosphatidylglycerol measurements are considerably less sensitive than the L:S ratio. The authors' results suggest that the immunologic test is probably useful as a screening test, but all negative results should be immediately followed up with an L:S ratio and phosphatidylglycerol measured by thin-layer chromatography.

The influence of relative body weight on toxicity of combination chemotherapy with cisplatin and etoposide.

PURPOSE: This study was conducted to determine whether there was any relationship between the adverse toxicity of combination chemotherapy and clinical values including age, sex, creatinine clearance (Ccr), body surface area and relative body weight. METHODS: Cisplatin at a dose of 80 mg/m2 on day 1 and etoposide at a dose of 100 mg/m2 on days 1, 2 and 3 were given to 42 consecutive patients with solid tumors. All patients had normal major organ function and received uniform hydration therapy. RESULTS: Body Mass Index as a measure of relative body weight was inversely correlated with the percentage decrease in white blood cells (P = 0.0681) and platelet count (P = 0.0115). Body surface area was also inversely correlated with leukopenia (P = 0.0171) and thrombocytopenia (P = 0.0058). In contrast, age, sex and Ccr had no significant relationship with adverse toxicity. CONCLUSIONS: It is concluded that dose adjustment of combination chemotherapy with cisplatin and etoposide according to age or ideal body weight is not appropriate and that a conventional dose modification method based solely on body surface area is probably not sufficient to reduce interpatient variability of cancer chemotherapy. A pharmacokinetic and pharmacodynamic study of combination chemotherapy is warranted to establish the ideal dose modification method.

A prognostic-factor risk index in advanced non-small-cell lung cancer treated with cisplatin-containing combination chemotherapy.

Prognostic factors for response and survival were retrospectively evaluated in 192 previously untreated patients with advanced non-small-cell lung cancer (NSCLC) who had received either vindesine plus cisplatin or mitomycin plus vindesine plus cisplatin as initial treatment. Univariate analysis demonstrated that squamous-cell histology, early stage, and a small number of metastatic sites were favorable prognostic factors for response to chemotherapy. Multivariate analysis using Cox's proportional hazard model indicated that the number of metastatic sites was the only significant pretreatment factor for response (P = 0.0005). Multivariate regression analysis revealed that the number of metastatic sites (P = 0.0002), sex (P = 0.0009), serum albumen levels (P = 0.0018), performance status (P = 0.0026) and lactic dehydrogenase values (P = 0.0026) contributed independently to survival. On the basis of these five prognostic factors, a prognostic index for survival was used to define three prognostic groupings (good, intermediate, and poor) for survival (median survival, 16.5 vs 9.4 vs 4.6 months; P = 0.0001). This particular regression model should aid in the design and analysis of new treatment strategies and may be useful for indirect comparisons of different studies carried out in similar patient populations.

Frequent allelic loss/imbalance on the long arm of chromosome 21 in oral cancer: evidence for three discrete tumor suppressor gene loci.

Frequent allelic imbalances including loss of heterozygosity (LOH) and microsatellite instability (MI) on the long arm of chromosome 21 (21q) have been found in several types of human cancer. This study was designed to identify tumor suppressor locus (or loci) associated with oral squamous cell carcinoma (SCC) on 21q. Among 38 patients with oral SCC tested, 15 (44%) of 34 informative cases showed LOH at one or more loci. Deletion mapping of these 15 tumors revealed three discrete commonly deleted regions on the chromosome arm. A minimal region with frequent LOH was found at the marker D21S236 mapped on 21q11.1. Another region of frequent deletion was identified between markers D21S11 and D21S1436 on 21q21, and a further commonly deleted region was found at D21S1254 on 21q22.1. In addition, we have detected MI on the chromosome arm in our oral SCC samples with significant correlation with tumor stage. Thus, our results strongly suggest that allelic imbalances on 21q may be involved in the development of oral SCC; and that at least three different putative tumor suppressor genes contributing to the pathogenesis of this disease are present on 21q.

Evidence of multi-step oncogenesis of oral squamous cell carcinoma.

We examined biopsy samples from one oral cancer and three precancerous lesions of the tongue of an 81-year old woman by polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequence analyses using 18 oligonucleotide primer pairs of adenomatous polyposis coli (APC) gene and 5 primers of p53 gene. Normal tongue epithelium adjacent to lesions was used as a control. The four lesions harbored the common mutation of APC gene that was not detected in the control. At codon 1621 in exon 15 of the APC gene there was a C to G substitution resulting in serine (TCA) to stop codon (TGA). No mutation of p53 gene was detected in any samples of the control and three precancerous lesions of the tongue. On the other hand, an A to G substitution at codon 170 in exon 5 of p53 gene resulting in glutamic acid (ACG) to glycine (GCG) was detected in the DNA of her tongue cancer. These results may suggest that the four lesions have the same origin, and that multi-step oncogenesis had occurred, the APC gene alteration being one of the early events in the process of tumorigenesis and p53 gene alteration involved in the late events.

Allelic loss on the short arm of chromosome 8 in oral squamous cell carcinoma.

Allelic deletions on the short arm of chromosome 8 (8p) are frequent events in many different types of malignant tumors, including head and neck tumors and oropharyngeal cancers. These regions are thought to harbor tumor suppressor genes. However, there has been no detailed analysis of loss of heterozygosity (LOH) on the chromosome arm 8p in oral squamous cell carcinoma (SCC). In order to estimate details of 8p loci involved in oral SCC, 32 patients with oral SCCs were examined for the LOH state 8p by PCR-LOH assay using 14 microsatellite markers. Based on our results a detailed deletion map of 8p showed LOH in at least one of the loci tested in 62.5% (20 of 32) of patients; and microsatellite instability (MI) was observed in 50% (16 of 32). The frequent LOH on this chromosome arm was identified at D8S87 and D8S258, mapped on 8p12 and 8p22, respectively. Fisher's exact test revealed no significant statistical correlation between the incidence of LOH or MI and clinicopathological features. Our observations indicate that the short arm of chromosome 8 may play a role in the pathogenesis of oral SCC; and that the two loci identified in this study may be tumor suppressor gene loci on 8p.

Dose-related alterations in growth and mineral disposition by chronic oral cadmium administration in the male rat.

The effect of cadmium (1, 10, or 100 ppm) administered to male rats in drinking water for 13 weeks on body weight and mineral disposition (Cd, Mg, P and Zn) in several body tissues was examined. Most alterations observed in these parameters occurred only at the 100 ppm dose of Cd. Terminal body weight was decreased by 10% in rats ingesting Cd at 100 ppm resulting from decreased food intake since weight gain/food consumption ratio was the same for all treatment groups. In serum, cadmium ingestion resulted in an inhibition of alkaline phosphatase activity at all concentrations and phosphorous was elevated only in animals receiving 100 ppm Cd. No changes were observed in Ca in urea. In bone, Cd decreased zinc content, increased Ca content, but did not influence bone ash, Mg or P and roentgenographic examination revealed no bone abnormalities. In both liver and kidney, cadmium ingestion did not influence intestinal absorption of Ca, Mg, P, or Zn or the renal excretion of Ca, P, or urea. The results of this study indicate that alterations in body weight and tissue mineral disposition resulting from chronic Cd ingestion are dose-related.

Sex-related differences in cadmium-induced alteration of drug action in the rat.

3 days after pretreatment of rats of both sexes with cadmium (2 mg/kg, 1.p.) the duration of hypnosis induced by hexobarbital (75 mg/kg, 1.p.) was potentiated in males but not females. Likewise similar treatment with cadmium leads to significant inhibition of the metabolism of hexobarbital by hepatic microsomal enzymes obtained from male but not female animals. These data suggest that there is a sex-related difference in the ability of cadmium to alter drug action in rats.

Establishment of a new human megakaryoblastic cell line, CMY, with chromosome 17p abnormalities.

A new megakaryoblastic cell line CMY was established from a Down's syndrome patient suffering from acute megakaryoblastic leukemia. The karyotypes of CMY showed deletion of chromosome 17 or the translocation of 17p, whereas the blasts of the patient did not reveal these abnormalities of chromosome 17 by conventional karyotype analysis. Blasts of the patient failed to respond to chemotherapy and complete remission could not be attained. The abnormalities of 17p became progressively predominant in the patient. These results suggest that the blasts of a minor clone which had the abnormalities of chromosome 17p might have existed in the patient from the beginning and CMY was established from the minor clone. Investigation of p53 gene by PCR-SSCP analysis revealed that blasts of the patient showed normal patterns, while CMY showed an abnormally migrating band in exon 5 alone. This result suggests that another novel oncogenic factor(s) besides p53 might be present on chromosome 17p and other tumor suppresser genes need to be studied.

Localization of a novel tumor suppressor gene loci on chromosome 9p21-22 in oral cancer.

Allelic imbalance or loss of heterozygosity (LOH) studies have been used to identify regions on chromosomes that may contain putative tumor suppressor genes. Deletions of chromosome 9 regions have been observed at high frequency in many other types of sporadic tumor, whereas in oral cancer no decisive information about the allelic loss on chromosome 9 has been reported. To provide detailed understanding of the genetic alterations in oral cancer, 24 highly polymorphic markers mapped on chromosome 9 were used to examine 34 cases of oral squamous cell carcinoma (SCC). LOH was detected in 18 (53%) of 34 informative samples at one or more loci examined. On the basis of our results, three commonly deleted regions were identified and a detailed deletion map was constructed. One of the novel regions was on 9p22, where a tumor suppressor gene, interferon a cluster (IFNA) gene, was identified before. Another region was D9S157 locus at 9p22, telomeric to IFNA locus and p15/16 genes, and the third was located on 9p21 of the D9S104 locus, centromelic to methylthioadenosine phosphorylase (MTAP) gene and p15/16 genes. Thus, our data suggest that, except for p15/16 and MTAP gene, there were at least two candidate tumor suppressor genes located at chromosome 9p, and that the alteration of these genes is associated with the tumorigenesis of oral SCC.

Gingival squamous cell carcinoma in a patient with chronic graft-versus-host disease.

This article describes a gingival squamous cell carcinoma that developed in a 21-year-old woman who received a bone marrow transplant at the age of 16 from her human leukocyte antigen-identical sister as treatment for severe aplastic anemia. Thirty days after transplantation, she presented with cutaneous erythema as a result of acute graft-versus-host disease, and this subsequently evolved into chronic graft-versus-host disease. A lichenoid white plaque of the gingiva developed shortly thereafter, and it began to increase in size rapidly 4 years posttransplantation. Biopsy indicated squamous cell carcinoma arising in this region, apparently associated with chronic graft-versus-host disease. Few reports have described a secondary solid malignancy involving the oral cavity of young adults after bone marrow transplantation.

Acquisition and transfer of sentence construction in autistic students: analysis by computer-based teaching.

In this study, we examined the conditions necessary to construct appropriate sentences in three autistic students using computer-based training and testing procedure. In Experiment 1, when a picture was presented on the computer display as a sample stimulus, the student was required to construct an appropriate sentence with five words. After training with three stimuli, each student could construct the correct sentence for 24 untrained stimuli. Appropriate vocal responses also emerged. In Experiment 2, the appropriate use of particles, which specify the subject and the object, was acquired by particle choice or sentence construction training. The rule was transferred to untrained stimuli and writing response. These results are discussed in terms of applicability of computer-based training for establishing appropriate sequential responding and particle usage in autistic students.

Ofloxacin concentrations in serum, saliva and pleural effusion of patients with pulmonary tuberculosis and lung cancer.

The ofloxacin (OFLX) concentration in serum, saliva and pleural effusion was measured in 12 patients with pleural effusion after oral administration at a dose of 200 mg three times a day (600 mg daily). Three patients had non-small cell lung cancer and the others had pulmonary tuberculosis. The mean OFLX levels in the serum, saliva and pleural effusion at 2 hours after the first administration on day 3 was 3.15 +/- 1.52, 3.36 +/- 2.23 and 2.86 +/- 1.77 micrograms/ml respectively. There was a strong correlation among these concentrations. The OFLX concentration of pleural effusion was predictable from that of saliva. A 3-day oral administration is sufficient to achieve the OFLX level of pleural effusion similar to that of the serum. It is possible that OFLX is effective for pleuritis caused not only by common infectious pathogens but also by Mycobacterium tuberculosis.