Occurrence of mutations in the epidermal growth factor receptor gene in X-ray-induced rat lung tumors

Epidermal growth factor receptor ( EGFR ) gene alterations have been found in human lung cancers. However, there is no information on the factors inducing EGFR mutations. In rodents, K- ras mutations are frequently found in many lung carcinogenesis models, but hitherto, Egfr mutations have not been reported. Their presence was therefore investigated in representative lung carcinogenesis models with 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), N -nitrosobis(2-hydroxypropyl)amine (BHP), 2-amino-3,8-dimethylimidazo[4,5- f ]quinoxaline (MeIQx) and ethyl carbamate (urethane), as well as X-ray irradiation. With the chemical carcinogenesis models, no mutations were detected in Egfr , which is in clear contrast to the high rates observed in either codon 12 or 61 of K- ras (21/23 of the lung tumors induced with NNK, 4/5 with MeIQx, 1/4 with urethane and 7/18 with BHP). However, in the X-ray-induced lung tumors, Egfr mutations with amino acid substitution were observed in exons 18 and 21 (4/12, 33%), but no activating mutation of K- ras was detected. In addition, one and four silent mutations were identified in K- ras (exon 1) and Egfr (exons 18, 20 and 21), respectively. Most mutations in both Egfr and K- ras were G/C→A/T transitions (7/8, 88% and 31/34, 91%, respectively). Although, the mutational patterns in equivalent human lesions were not completely coincident, this first report of Egfr mutations in an experimental lung tumor model suggests that X-rays or other factors producing oxygen radicals could cause EGFR mutations in some proportion of lung cancers in humans. ( Cancer Sci 2008; 99: 241–245)     

Midkine and its clinical significance in endometrial carcinoma

Midkine (MK) is a secreted heparin-binding growth factor. Several types of human cancer have increased MK expression with elevated serum levels. The purpose of this study was to determine whether MK was expressed in endometrial carcinoma and to evaluate the clinicopathological significance of serum MK in patients with endometrial carcinoma. Immunohistochemical expression of MK was evaluated in 85 endometrial carcinoma samples and 33 controls. MK expression was significantly higher in the carcinomas than in normal endometrium ( P <  0.001). Interestingly, MK expression was highest at the margins of invasion and low in the superficial areas of the tumor samples. Using ELISA, we compared serum MK concentration in 120 endometrial carcinoma patients with the concentration in 46 patients with benign gynecologic tumors. Serum MK value in patients with cancer was significantly higher than that in the patients with benign diseases ( P  = 0.01). Patients with positive lymph node metastasis or recurrence, or cancer death, had a higher serum MK level ( P =  0.008, P  = 0.009, respectively). In conclusion, MK immunoreactivity in endometrial carcinoma is significantly higher than in normal endometrium. Additionally, preoperative serum MK levels are significantly correlated with prognosis and the presence of lymph node metastasis. Thus, MK may be a useful serum biomarker for identifying high risk patients of endometrial carcinoma. ( Cancer Sci 2008; 99: 1125–1130)     

Prospective evaluation of selection criteria for active surveillance in Japanese patients with stage T1cN0M0 prostate cancer

Objective: Selection criteria for active surveillance (AS) program of localizedprostate cancer remain to be standardized. The purpose was toevaluate the validity of selection criteria and investigatethe feasibility of this AS program. Methods: Patients meeting the criteria (i) stage T1cN0M0, (ii) age 50–80,(iii) serum prostate-specific antigen (PSA) 20 ng/ml, (iv) oneor two positive cores per 6–12 systematic biopsy cores,(v) Gleason score 6, and (vi) cancer involvement in positivecore 50% were enrolled and encouraged to start AS for at least6 months during the period between January 2002 and December2003. PSA was measured bimonthly for 6 months and every 3 monthsthereafter. Trigger of treatment recommendation was PSA-doublingtime (PSADT) of 2 years or pathological progression at re-biopsy.Primary endpoint was ‘%PSADT > 2y’, which wasdefined as the proportion of patients who showed PSADT assessedat 6 months >2 years out of all the patients who chose AS.Point estimate of ‘%PSADT > 2y’ was expectedto be >80%. Results: One hundred and eighteen patients opted for AS and 16 choseimmediate treatment at enrollment. PSADT for the initial 6 monthsbased on four measurements could be assessed in 106 patients.Intent-to-treat analysis of ‘%PSADT > 2y’ was71.2% (84/118, 95% CI: 62.1–79.2). Pathological progressionrate at 1-year re-biopsy was 33%. Fifty-four (46%) patientsremained on AS for maximal observation of 54 months. Generalhealth-related QOL in patients undergoing AS was not impaired. Conclusions: The primary endpoint, ‘%PSADT > 2y’, did notmeet the pre-specified decision criteria. Further prospectivestudy with revised program and endpoint is needed.     

Clinicopathological and prognostic relevance of uptake level using 18F-fluorodeoxyglucose positron emission tomography/computed tomography fusion imaging (18F-FDG PET/CT) in primary breast cancer

Objective: Using integrated ¹⁸F-fluorodeoxyglucose positron emission tomography/computedtomography fusion imaging (¹⁸F-FDG PET/CT), the clinical significanceof ¹⁸F-FDG uptake was evaluated in patients with primary breastcancer. Methods: Clinicopathological correlation with the level of maximum standardizeduptake values (SUV) 60 min obtained from preoperative ¹⁸F-FDGPET/CT were examined in 152 patients with primary breast cancer.The prognostic impact of the level of SUV was explored usingsimulated prognosis derived from computed program Adjuvant!in 136 (89%) patients with invasive ductal carcinoma (IDC). Results: High SUV level was significantly correlated with tumor invasivesize (2 cm) (P <0.0001), higher score of nuclear grade (P<0.0001), nuclear atypia (P <0.0001) and mitosis counts(P <0.0001), negative hormone receptor status (P = 0.001),high score of c-erbB-2 expression (P = 0.006), lymph node metastasis(P = 0.002), and IDC in comparison with invasive lobular carcinoma(P = 0.004). Multivariate analyses showed tumor invasive size,nuclear grade and estrogen receptor negativity were significantlycorrelated with SUV in primary breast cancer (P <0.0001,<0.0001,and <0.012, respectively), and nuclear grade was significantlycorrelated with SUV in tumors of invasive size 2 cm or less(P <0.0001). Tumors with high SUV (cutoff value 4.0) showedhigher relapse and mortality rate compared to those with lowSUV (P <0.0001). Conclusions: High uptake of ¹⁸F-FDG would be predictive of poor prognosisin patients with primary breast cancer, and aggressive featuresof cancer cells in patients with early breast cancer. ¹⁸F-FDGPET/CT could be a useful tool to pretherapeutically predictbiological characteristics and baseline risk of breast cancer.     

Increased expression of germinal center-associated nuclear protein (GANP) is associated with malignant transformation of melanocytes

BACKGROUND: Germinal center-associated nuclear protein (GANP) is a newly cloned molecule that is up-regulated in the germinal center B cells. Although GANP functions in the regulation of DNA repair during replication and survival of B cells, little is known about its expression in melanocytic cells. OBJECTIVES: To investigate whether GANP and phosphorylated-GANP (P-GANP) are expressed in cultured human melanocytes and melanoma cells and in benign and malignant melanocytic lesions. In addition, we aim to determine whether GANP and P-GANP are associated with malignant transformation of melanocytic lineage. METHODS: GANP and P-GANP expression in cultured melanocytic cells was analyzed by immunostaining and in vitro kinase assay. GANP and P-GANP expression in melanocytic lesions was analyzed by immunohistochemistry. RESULTS: GANP and P-GANP were up-regulated in cultured melanoma cells compared to melanocytes. GANP and P-GANP were restricted to nucleus of melanocytes but co-expressed in cytoplasm of melanoma cells. On the other hand, GANP and P-GANP were widely expressed at various levels in melanocytic nevi and melanoma lesions with nuclear and cytoplasmic staining pattern. Melanoma cells showed a stronger intensity of GANP and P-GANP than melanocytic nevus cells, however the staining intensity in primary melanoma lesions was not associated with any clinicopathological variables. Cytoplasmic GANP and P-GANP expression was associated with MCM3 and Ki67 expression. CONCLUSIONS: These data suggest, for the first time, that GANP and P-GANP are up-regulated in cultured melanoma cells compared to melanocytes and also they are widely expressed in benign and malignant melanocytic tumor cells.     

Comparison of the protein composition of breast milk and the nutrient intake between Thai and Japanese mothers

This study compared the protein composition of breast milk and the nutrient intake between Thai and Japanese lactating mothers. The breast milk was collected from 15 Thai and 14 Japanese mothers at the fifth day post-partum. Twenty-four-hour dietary records were performed from the second-to-the-fourth day post-partum. The nutrient intake was calculated by using the nutrient content of a food table. The protein composition of the whey was separated by gel electrophoresis and was identified by mass spectrometry and two-dimensional electrophoresis. The results showed that the concentrations of the major protein types in the breast milk were not significantly different between the two groups. The concentrations of the minor protein types varied markedly with the individuals, with higher concentrations in the breast milk of the Thai mothers. There were no significant differences in terms of the energy and protein intake; however, the sources of energy were different. The results indicate that the total protein and lactoferrin concentrations in the breast milk could be predicted by the maternal daily energy and fat intake.     

Measurement of Serum 17α-hydroxyprogesterone in Newborn Infants by Stable Isotope Dilution—Gas Chromatography/Mass Spectrometry

Immunochemical measurement of serum 17α-hydroxyprogesterone (17OHP), the most important parameter for diagnosis of classical 21-hydroxylase deficiency (21OHD) in newborn infants, is known to be inaccurate due to the cross-reactivity of antibodies with a large quantity of fetal adrenal steroids. The aims of this study were 1) to establish reference values for the serum 17OHP level in Japanese newborn infants using non-immunochemical stable isotope dilution —gas chromatography/mass spectrometry (SID-GC/MS) and 2) to compare the serum 17OHP levels determined by SID-GC/MS with those determined by radioimmunoassay (RIA). The first study subjects were used for determination of reference values and included 57 healthy full-term newborn infants (4–5 d of age). The second study subjects were used for comparison of SID-GC/MS with RIA and included 27 healthy full-term newborn infants (3–6 d of age) and two subjects with neonatal transient hyper 17OHPnemia; these two subjects were 16 and 27 d of age, respectively. In the first study subjects, the intra-assay coefficient of variation for SID-GC/MS was 3% (n=5), the recovery rate was 98%, the sensitivity was 0.2 ng/ml, and the range of linearity was 0.5–200 ng/ml. The reference values for the serum 17OHP level determined by SID-GC/MS ranged from 0.3–1.5 (0.6) (ng/ml) (median). In the second study subjects, the serum 17OHP levels determined by SID-GC/MS were lower in one of the 27 subjects and both of the two subjects with neonatal transient hyper 17OHPnemia compared with the levels determined by RIA. Measurement of the serum 17OHP level using SID-GC/MS may be clinically useful for definitive diagnosis of classical 21OHD in newborn infants.     

Long term outcomes in patients with intracranial germinomas: a single institution experience of irradiation with or without chemotherapy

Complete remission can be achieved soon after irradiation in patients with intracranial germinoma. This study aimed to analyze the follow-up outcome of intracranial germinoma patients. About 39 intracranial germinoma patients (29 males and 10 females; average age, 15 years; range, 7–27 years) treated at Kyoto University Hospital from 1978 to 2004 were included in the study group. Six patients had multifocal disease at initial diagnosis, and 10 had human chorionic gonadotropin (HCG)-producing tumors. Thirteen patients were treated with craniospinal axis irradiation, 6 with whole-brain irradiation, 17 with whole-ventricle irradiation, and 3 with local field irradiation. Since 1997, 15 patients were treated with reduced–dose whole-ventricle irradiation (median, 23.4 Gy; range, 20.4–27 Gy) followed by a local boost (median, 40.8 Gy; range, 36–54 Gy) combined with chemotherapy. The median follow-up was 94 months (18 months to 25 years). The 5- and 10-year overall survival (OS) rates of the entire group were 97 and 90%, respectively. The 5- and 10-year progression-free survival (PFS) rates of the entire group were 91 and 87%, respectively. The 8-year OS and PFS in 15 patients treated by whole-ventricle irradiation combined with chemotherapy were 100% and 92%, respectively. Four patients had recurrences within a median period of 59.5 months (51–85 months). All relapses occurred outside the radiation fields. Tumor site, tumor size, HCG production, multifocal disease and radiation dose to the primary site or whole ventricle did not significantly affect PFS. All initial recurrences of intracranial germinoma occurred at the distant site out of the radiation field. Our data suggested that reduced doses to the whole ventricle, combined with chemotherapy, should be sufficiently effective in patients with intracranial germinoma.