Detection of Tumor DNA in Serum of Colorectal Cancer Patients

We previously found p16 promoter methylation in DNA in the sera of 13 colorectal cancer patients out of 44 (30%) whose tumor DNA exhibited the methylation, using methylation-specific PCR (MSP). To examine whether the cancer detection rate could be improved by using a different tumor marker, we examined the K- ras status in 90 colorectal cancer patients using a mismatch ligation assay. Among the 31 patients showing K- ras gene mutations in their tumors, the same mutations were observed in serum DNA of 11 patients (35%). Among the 90 patients, 63 showed tumors positive for K- ras mutation or p16 promoter methylation, or both, and 22 had serum DNA positive for one or both. K- ras mutation was found even in serum DNA of patients with Dukes A cancer, suggesting that colorectal cancer might be detected even in patients without symptoms by using this ligation assay.     

Serum Insulin-like Growth Factors, Insulin-like Growth Factor-binding Protein-3, and Risk of Lung Cancer Death: A Case-control Study Nested in the Japan Collaborative Cohort (JACC) Study

To elucidate the roles of insulin-like growth factors (IGFs) in the development of lung cancer, we conducted a case-control study nested within the Japan Collaborative Cohort Study. Serum samples were collected at baseline from 39 140 men and women between 1988 and 1990. We measured serum IGF-I, IGF-II, and IGF-binding protein-3 (IGFBP-3) in 194 case subjects who subsequently died from lung cancer during an 8-year follow-up and in 9351 controls. The odds ratios (ORs), adjusted for smoking and other covariates, were smaller with higher levels of IGF-II and IGFBP-3. The ORs across quartiles were 0.41 (95% confidence interval [CI], 0.27–0.63), 0.47 (0.31–0.71), and 0.67 (0.46–0.98) for IGF-II (trend P =0.018), and 0.55 (95% CI, 0.37–0.81), 0.54 (0.36–0.82), and 0.67 (0.45–1.01) for IGFBP-3 (trend P =0.037). These peptides were not independently related to lung cancer risk when mutually adjusted. The risk was increased in the highest vs. the lowest quartile of IGF-I only after controlling for IGFBP-3 (OR, 1.74; 95% CI, 1.08–2.81). Limiting subjects to those followed for ≥3 years strengthened the negative associations of IGF-II and IGFBP-3, whereas the ORs for IGF-I generally decreased. A higher level of circulating IGFBP-3 and/or IGF-II may decrease lung cancer risk. Elevated serum IGF-I may increase the risk, but this could partly be attributable to latent tumors.     

Inhibitory Effects of Toremifene on N-Methyl-N-nitrosourea and Estradiol-17β-induced Endometrial Carcinogenesis in Mice

Short- and long-term experiments were designed to determine the effects of toremifene (TOR) on estrogen-related endometrial carcinogenesis in mice. In the short-term experiment, a single low dose of TOR (0.2 mg/30 g body weight) decreased expression of c- fos , interleukin (IL)-1α, estrogen receptor (ER)-α mRNAs and corresponding proteins induced by estradiol-l7β (E₂), in the uteri of the ovariectomized mice. Expression of ER-β mRNA was increased by the TOR treatment, compared with the control. In the long-term experiment, 106 female ICR mice were given N -methyl N -nitrosourea (MNU) into their uterine corpora. The animals were divided into four groups as follows: group 1, E₂ diet (5 ppm) plus TOR (0.2 mg/30 g body weight, subcutaneously, every four weeks); group 2, E₂ diet alone; group 3, basal diet plus TOR. Group 4 served as the control. TOR treatment decreased the incidence of MNU and E₂-induced endometrial adenocarcinoma and atypical hyperplasia at the termination of the experiment (30 weeks after the start). These results suggest that TOR exerts preventive effects against estrogen-related endometrial carcinogenesis in mice, through the suppression of c- fos as well as IL-1α expression induced by E₂. Such suppressive effects of TOR may be related to the decreased ER-α and increased ER-β expressions.     

Human leukocyte antigen (HLA) phenotypes in siblings with osteosarcoma

Osteosarcoma was detected in two siblings. Their human leukocyte antigen (HLA) phenotypes were completely identical, although they were different from those of osteosarcoma patients in previous reports. Despite an extensive search of family and past history, no significant background related to the induction of cancer could be found. These cases suggest that genetic similarity may influence the development of osteosarcoma. Ascertainment of the HLA phenotypes in siblings with osteosarcoma might be a useful strategy to facilitate the early diagnosis of this tumor.     

Blood-pool scintigraphic diagnosis of fractured lumbar vertebral hemangioma

A 57-year-old woman complained of lumbago of 1 year’s duration. Radiographs showed a compression fracture of the third lumbar vertebra. CT and MR images revealed an enhancing mass confined to the vertebral body suggestive of a malignant process. A blood-pool scintigram with ^99mTc-human serum albumin combined with DTPA (HSA-D) revealed marked accumulation. This strongly suggested a hemangioma, which was confirmed by biopsy. Received: 26 October 2000 Revision requested: 21 November 2000 Revision received: 23 December 2000 Accepted: 28 December     

Effects of Preservation of Mouse Spermatozoa in Electrolyte-Free Solution at 4°C on the Outcome of Mouse In Vitro Fertilization

Purpose: The aim was to assess the fertilizing capacity ofspermatozoa cool-preserved in electrolyte-free (EF)solution. Methods: Mouse spermatozoa were cool-preserved in EFsolution and the acrosomal status of the spermatozoa wascompared before and after preservation using chlortetracyclinestain. Mouse oocytes were inseminated by spermatozoacool-preserved in EF solution for 2, 4, or 7 days and fertilizationand blastocyst rates were evaluated. Results: Acrosomal status of spermatozoa cool-preservedin EF solution was not different from spermatozoa beforepreservation, but the capacitated and acrosome-reactedspermatozoa significantly increased after reinitiation. Cool-preservationin EF solution for up to 4 days did not affectfertilization rate. Blastocyst rate of embryos derived fromspermatozoa cool-preserved for 4 or 7 days in EF solutionwas significantly lower than that of embryos derived fromfresh spermatozoa. Conclusions: Mouse spermatozoa cool-preserved in EFsolution possesses as much fertilizing capacity as fresh spermatozoa.However, prolonged preservation affects theembryonic development.     

Central Congenital Hypothyroidism Detected by Neonatal Screening in Sapporo, Japan (2000–2004): It’s Prevalence and Clinical Characteristics

In Sapporo, Japan, a neonatal screening program for congenital hypothyroidism (CH) has employed measurement of free thyroxine (T4) and TSH in the same filter-paper blood spot. This system has enabled us to identify primary CH and central CH during the neonatal period. The aim of this study was to clarify the prevalence and clinical characteristics of central CH. For this purpose, the screening program requested serum from infants with free T4 concentrations below the cut off value regardless of the TSH levels. Between January 2000 and December 2004, 83,232 newborns were screened and six central CH patients were detected as a result of follow-up of low free T4 and non-elevated TSH screening (1:13,872). This frequency is higher than in other studies. Four patients showed multiple pituitary hormone deficiency with pituitary malformations on magnetic resonance imaging. One patient was diagnosed as having Prader-Willie syndrome. The remaining patient was considered to have isolated central CH. Our study demonstrated that the frequency of central CH is 1:13,872. Free T4 measurement would also be advantageous in early recognition of multiple pituitary hormone deficiency.     

Case Report: Adjuvant Therapy with a High Dose of Mitotane for Adrenocortical Carcinoma in a 4-year-old Boy

This report concerns control of adrenocortical carcinoma in a 4-yr-old boy by adjuvant mitotane therapy. He presented precocious puberty and was diagnosed with adrenocortical carcinoma. He underwent surgical resection, and adjuvant mitotane therapy was initiated, leading to a final dose of 5.0 g/day. Despite monitoring of the plasma mitotane level, encephalopathy developed 5 mo after initiation. Although he recovered from the encephalopathy, careful follow-up of his growth and development is necessary. On the other hand, he has been free of recurrence and metastases for 3 yr since discontinuation of mitotane. A high dose of mitotane is potentially effective as an adjuvant chemotherapy for adrenocortical carcinoma, although optimal and safe usage needs to be established for children.     

Altered Expression of γ-Glutamylcysteine Synthetase, Metallothionein and Topoisomerase I or II during Acquisition of Drug Resistance to Cisplatin in Human Ovarian Cancer Cells

This study was designed to elucidate the mechanisms of cisplatin (CDDP) resistance using two human ovarian cancer cell lines, KF and TYK, and two CDDP-resistant lines, KFr and TYK/R, derived from the former lines. KFr and TYK/R showed about 3-fold higher resistance to the cytotoxic effects of CDDP than their parental lines. They also showed a significant increase in sensitivity to not only etoposide, but also (+)-(4S)-4, ll-diethyl-4-hydroxy-9-[(4-piperidino-piperidino)carbonyloxy]-l H -pyrano[3',4':6,7]inodolizino[l,2- b ]quinoline-3,14(4 H , 12 H )-dione hydrochloride trihydrate (CPT-11). Cellular CDDP accumulation levels in KFr and TYK/R were decreased from those of the parental cells. By contrast, the cellular glutathione (GSH) content in KFr cells was 1.7-fold higher than that in KF, whereas TYK/R cells had a 40% lower content than TYK cells. Cellular mRNA levels of drug-resistance-related genes, such as DNA topoisomerase (topo) I and topo II, glutathione S-transferase-π (GST-π;), γ-glutamylcysteine synthetase (.γ -GCS ), and metallothionein ( hMT ) genes, were compared between drug-sensitive KF or TYK and KFr or TYK/R. KFr cells had 8.5- and 24.7-fold higher mRNA levels of γ-GCS and topo II genes than KF cells while KFr had only a slight increase in GST-π mRNA level as compared with KF. By contrast, TYK/R cells had 2.9- and 1.7-fold higher hMT and topo I mRNA levels than TYK cells. Acquisition of CDDP resistance in human ovarian cancer cells thus appeared to be related mainly to expression of γ -GCS, topo II and hMT genes, and partly to that of topo I and GST-π genes, in addition to a decrease in CDDP accumulation