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Monopolar spindle 1 (Mps1) is an attractive cancer drug target due to the important role that it plays in centrosome duplication, the spindle assembly checkpoint, and the maintenance of chromosomal stability. A design based on JNK inhibitors with an aminopyridine scaffold and subsequent modifications identified diaminopyridine 9 with an IC50 of 37 nM. The X-ray structure of 9 revealed that the Cys604 carbonyl group of the hinge region flips to form a hydrogen bond with the aniline NH group in 9. Further optimization of 9 led to 12 with improved cellular activity, suitable pharmacokinetic profiles, and good in vivo efficacy in the mouse A549 xenograft model. Moreover, 12 displayed excellent selectivity over 95 kinases, indicating the contribution of its unusual flipped-peptide conformation to its selectivity.

Objectives: This study was undertaken to measure the radiation exposure level of caregivers following outpatient NaI (I-131) 1,110 MBq therapy for remnant thyroid ablation after total thyroidectomy in patients with differentiated thyroid cancer, and to evaluate the influence of activities of daily living on radiation exposure level, with the goal of proposing an optimum method of I-131 therapy., Methods: The study included 37 patients with differentiated thyroid cancer, who had undergone total thyroidectomy and received outpatient based remnant thyroid ablation using NaI (I-131) 1,110 MBq, who were satisfying the following requirements: (1) patients who have no evidence of distant metastases, (2) whose living environments were appropriate for outpatient I-131 (1,110 MBq) therapy, and (3) patients who gave written informed consent. The dose rate at a distance of 1 m from the body surface of the patient at the moment of release was measured using survey meters of the GM type or ionization chamber type. The dose level for the caregiver was measured with a personal dosimeter in all cases., Results: The dose rate at a distance of 1 m from the patient's body surface 1 h after I-131 administration was in the range of 29-115 μSv/h (mean 63.8 μSv/h). The 7-day cumulative effective dose of caregivers was 0.11 ± 0.08 mSv, on an average, in 34 dosimeters. In 31 of 34 dosimeters, cumulative effective dose of caregivers was below 0.2 mSv. Dose levels exceeding 0.2 mSv were recorded in 3 cases (0.21, 0.35 and 0.43 mSv in one case each). These results suggest that the exposure level of family members (caregiver and others) was minimal and is lower than the radiation levels affecting human environments., Conclusion: Outpatient-based remnant thyroid ablation with I-131 (1,110 MBq) performed after total thyroidectomy in patients with differentiated thyroid cancer is safe if applied in accordance with the appropriate supervision and guidance by experts with certain qualifications.

Membrane-anchored receptor-like protein kinases (RLKs) recognize extracellular signals at the cell surface and activate the downstream signaling pathway by phosphorylating specific target proteins. We analyzed a receptor-like cytosolic kinase (RLCK) gene, ARCK1, whose expression was induced by abiotic stress. ARCK1 belongs to the cysteine-rich repeat (CRR) RLK sub-family and encodes a cytosolic protein kinase. The arck1 mutant showed higher sensitivity than the wild-type to ABA and osmotic stress during the post-germinative growth phase. CRK36, an abiotic stress-inducible RLK belonging to the CRR RLK sub-family, was screened as a potential interacting factor with ARCK1 by co-expression analyses and a yeast two-hybrid system. CRK36 physically interacted with ARCK1 in plant cells, and the kinase domain of CRK36 phosphorylated ARCK1 in vitro. We generated CRK36 RNAi transgenic plants, and found that transgenic plants with suppressed CRK36 expression showed higher sensitivity than arck1-2 to ABA and osmotic stress during the post-germinative growth phase. Microarray analysis using CRK36 RNAi plants revealed that suppression of CRK36 up-regulates several ABA-responsive genes, such as LEA genes, oleosin, ABI4 and ABI5. These results suggest that CRK36 and ARCK1 form a complex and negatively control ABA and osmotic stress signal transduction., (© 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd.)

Objectives: The purpose of this study was to estimate the diagnostic accuracy of (123)I-metaiodobenzylguanidine (MIBG) scintigraphy to diagnose Lewy body disease (LBD), including Parkinson's disease (PD) and dementia with Lewy bodies, and to clarify the relationship between MIBG parameters and the clinical findings., Methods: One hundred-and-forty-four patients with parkinsonism without diabetes mellitus or a history of cardiac disease were retrospectively selected in the study. Clinical diagnosis was confirmed by follow-up during more than 6 months by neurologists. All patients underwent MIBG imaging at 15 min (initial) and 4 h (delayed) after the tracer injection, and clinical features such as Hoehn and Yahr (H-Y) classification or symptoms specific to parkinsonism were also investigated. The heart to mediastinum ratio (H/M) and the washout ratio (WR) of MIBG were calculated, and correlation with the clinical features was analyzed., Results: Ninety-seven and 47 patients were diagnosed as LBD and Parkinson's syndrome (PS), respectively. Initial and delayed H/M were significantly lower and WR was significantly higher in LBD than in PS (p < 0.0001). The initial H/M was independently correlated with tremor (F value 10.45), hesitation (F = 4.49), and hallucinations (F = 5.09) (p < 0.0001). The sensitivity and specificity for the diagnosis of LBD were 64.9 and 87.2% with initial H/M, 78.4 and 68.1% with delayed H/M, and 80.4 and 61.7% with WR, respectively. Using multivariate analysis, initial H/M (F = 39.33) and tremor (F = 10.46) were independently correlated to the diagnosis of LBD (r = 0.562, p < 0.0001) among the MIBG and various clinical parameters., Conclusion: The initial H/M was the most useful of the 3 different parameters of MIBG for the diagnosis of LBD, but had low sensitivity. WR and delayed H/M had no incremental value to initial H/M for the diagnosis of PD. Careful long-term follow-up is needed for patients with parkinsonism who are clinically diagnosed as LBD with normal initial H/M, or diagnosed as no LBD with low initial H/M.

Esterification of oleic acid was performed in a three-phase fixed-bed reactor with a cation exchange resin catalyst (Amberlyst-15) at high temperature, which was varied from 80 to 120 °C. The fatty acid methyl ester (FAME) yields in the fixed-bed reactor were increased with increases in the reaction temperature, methanol flow rate and bed height. Moreover, the FAME yields were higher than those obtained using a batch reactor due to an equilibrium shift toward the product that resulted from continuous evaporation of the produced water. In addition, there was no catalyst deactivation during the esterification of oleic acid. However, addition of sunflower oil to the oleic acid reduced the FAME yield obtained from simultaneous esterification and transesterification. The FAME yield was 97.5% at a reaction temperature of 100 °C in the fixed-bed with a height of 5 cm when the methanol and oleic acid feed rates were 8.6 and 9.0 mL/h, respectively., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Objective: This study sought to assess the safety, efficacy, impact on hypothyroid symptoms, and pharmacokinetics of SKG-02 (rhTSH, thyrotropin alfa) in the diagnostic follow-up of Japanese patients with well-differentiated thyroid carcinoma (WDTC)., Methods: Ten Japanese adults with WDTC were enrolled into a prospective, multicenter, open-label trial comparing diagnostic whole-body scintigraphy (dxWBS) and serum thyroglobulin (Tg) testing aided by SKG-02 versus these procedures aided by thyroid hormone withdrawal (THW). Patients were their own controls. Variables compared included scan set ability to detect radioiodine uptake by remnant or malignant thyroid tissue, scan set quality, diagnostic sensitivity of dxWBS and Tg testing alone or combined, frequency of hypothyroid signs/symptoms, and adverse events (AEs). SKG-02 pharmacokinetic variables including maximum concentration (Cmax), time to Cmax (Tmax) and the area under the time-concentration curve (AUC) were calculated., Results: In a blinded evaluation by an independent committee of 3 nuclear medicine experts, 70% of SKG-02 dxWBS scan sets were rated "equivalent" (n = 7) or "superior" (n = 0) to their THW counterparts in ability to detect radioiodine uptake in healthy or malignant thyroid tissue. Therefore the study exceeded its primary endpoint of a 60% equivalence/superiority rate. SKG-02 Tg testing identified 3/3 cases of disease. Hypothyroid signs/symptoms were substantially more frequent during THW than during euthyroidism permitted by SKG-02 use. SKG-02 was well-tolerated, with no severe or serious drug-related AEs. Cmax was 240.8 +/- 65.9 microIU/ml, Tmax was 28.75 +/- 14.21 hr after the first SKG-02 injection, and AUC was 11,414 +/- 3,462 microIU hr/ml in 9 patients evaluable for pharmacokinetics., Conclusions: SKG-02 was safe and effective in the diagnostic follow-up of Japanese patients with WDTC, avoiding hypothyroid morbidity relative to THW. These and the pharmacokinetic findings were similar to those of overseas Phase III studies.

Purpose: Myocardial mitochondria are the primary part of energy production for healthy cardiac contraction. And mitochondrial dysfunction would play an important role in progressive heart failure. In the recent years, myocardial washout of (99m)Tc-sestamibi [((99m)Tc-hexakis-2-methoxy-2-methylpropyl isonitrile (MIBI)] has been introduced to be a potential marker in patients with heart failure. The objective of this study was to clarify MIBI extraction and washout kinetics using isolated perfusion system in hypertension induced model of myocardial dysfunction., Methods: Six-week-old Dahl-salt sensitive rats, allotted to 4 groups; a 5-week high-salt group (5 wk-HS), 12-week high-salt group (12 wk-HS) and two age-matched, low-salt diet control groups (5 wk-LS and 12 wk-LS). The rats in 5 wk-HS and 12 wk-HS groups were fed a high-salt diet (containing 8% NaCl). Cardiac function was examined by echocardiography before removing heart. Hearts were perfused according to the Langendorff method at a constant flow rate, in which 20-min MIBI washin was conducted followed by 25-min MIBI washout. Whole heart radioactivity was collected every sec by an external gamma detector. The myocardial extraction, K(1) (ml/min) and washout rate, k(2) (min(-1)) were generated., Results: High-salt diet groups showed significant high-blood pressure. Echocardiography revealed thickened LV walls in 5 wk-HS, and reduced cardiac function in 12 wk-HS, compared to each age-matched control group. K(1) showed no significant difference among all groups (5 wk-HS: 2.36 ± 1.07, 5 wk-control: 2.59 ± 0.28, 12 wk-HS: 1.91 ± 0.90, and 12 wk-control: 2.84 ± 0.57). k(2) in 5 wk-HS was comparable to that in the age matched control group (0.00030 ± 0.00039 vs -0.000010 ± 0.00044), but it was increased remarkably in 18 wk-HS compared to the age matched control group (0.0025 ± 0.0011 vs 0.000025 ± 0.000041, P<.01), and 5 wk-HS (P<.01)., Conclusion: In the course of hypertensive heart disease, MIBI washout was increased in the transitional state from hypertrophied to dilated and failing heart, while MIBI extraction remained intact., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Objective: Elevated levels of serum carcinoembryonic antigen (CEA) in patients with colorectal cancer (CRC) during follow-up suggest recurrence, which can be visualized by (18)F-FDG PET/CT. Since the magnitude of CEA elevation reflects cancer volume, the ability of PET/CT to detect recurrence in patients with only mildly elevated CEA might be limited. However, the accuracy of PET/CT in detecting recurrence associated with elevated CEA has not been fully assessed. We retrospectively evaluated the diagnostic performance of (18)F-FDG PET/CT postoperatively relative to CEA levels among patients with CRC., Methods: We visually assessed 75 PET/CT evaluations of 57 postoperative patients with CEA >5.0 ng/ml. Tumor volumes were also determined using image analysis software. The final diagnosis was confirmed based on histopathological findings, or at least on 6 months of clinical follow-up., Results: Two lung cancers were excluded and we finally analyzed data from 73 of the 75 PET/CT evaluations. Recurrences were diagnosed in 54 (prevalence 74%). The sensitivity and specificity of PET/CT to detect recurrence was 50/54 (93%) and 14/19 (74%), respectively. The positive and negative predictive values were 91 and 78%, respectively, and the positive and negative likelihood ratios were 3.52 and 0.10, respectively. Values for the sensitivity of PET/CT were 88 and 95%, and those for specificity were 78 and 70%, at serum CEA concentrations of 5-10 and >10 ng/ml, respectively. Serum CEA (r = 0.500, p < 0.001) significantly correlated with cancer volumes., Conclusions: The present findings showed that (18)F-FDG PET/CT could accurately detect recurrent CRC irrespective of the elevated CEA concentration.

Stable oxygen sites on a PdO film over a Pd(100) thin structure with a [Formula: see text] surface unit cell are determined using the first-principles electronic structure calculations with the generalized gradient approximation. The adsorbed monatomic oxygen goes to a site bridging two twofold-coordinated Pd atoms or to a site bridging a twofold-coordinated Pd atom and a fourfold-coordinated Pd atom. Estimated reaction energies of CO oxidation by reduction of the oxidized PdO film and N(2)O reduction mediated by oxidation of the PdO film are both exothermic. Motion of the adsorbed oxygen atom between the two stable sites is evaluated using the nudged elastic band method, where an energy barrier for a translational motion of the adsorbed oxygen may become ∼0.45 eV, which is low enough to allow fluxionality of the surface oxygen at high temperatures. The oxygen fluxionality is allowed by the existence of twofold-coordinated Pd atoms on the PdO film, whose local structure has a similarity to that of Pd catalysts for the Suzuki-Miyaura cross-coupling. Although NO(x) (including NO(2) and NO) reduction is not always catalyzed by the PdO film only, we conclude that continual redox reactions may happen mediated by oxygen-adsorbed PdO films over a Pd surface structure, when the influx of NO(x) and CO continues, and when the reaction cycle is kept on a well-designed oxygen surface.

Objective: A mismatch defect between (201)TL and (123)I-BMIPP dual isotope SPECT (d-SPECT) is useful to detect myocardial ischemia in patients with acute coronary syndrome. However, whether mismatched d-SPECT findings reflect actual myocardial ischemia in stable patients with suspected, but unknown ischemic heart disease is unclear. The present study assesses the significance of a d-SPECT mismatch among such patients., Methods: Forty-nine patients with suspected stable coronary heart disease who had been referred for chest pain, ECG abnormalities or multiple risk factors (66 +/- 11 years old, 34 males) with a d-SPECT mismatch participated in this study. All of them underwent coronary angiography (CAG) to assess coronary artery disease. The entire myocardial area on d-SPECT images was divided into 17 segments, each of which was scored from 0 (normal) to 4 (defect). The d-SPECT mismatch score (MS) was defined as the summed BMIPP defect score (BM-TDS) minus the summed defect score (TL-TDS). The inclusion criterion was MS >or= 1, and the mismatch was defined as true positive if the mismatched area was concordant with the territories supplied by significant coronary stenotic arteries by CAG., Results: Ischemic heart disease was judged by coronary angiography in 31 (63%) patients (IHD group), of which 24 (49.0%) were true positives. Of the remaining 18 (37%) patients without no significant coronary stenosis (non-IHD group), 12 (24%) had some types of organic heart disease. If MS >or= 4 was defined as the threshold for an ischemic positive mismatch, then the sensitivity and specificity were 80% and 63%, respectively. However, mismatch scores did not significantly differ between the groups with true positive-IHD and organic heart disease in non-IHD group (6.6 +/- 4.4 vs. 6.4 +/- 3.7)., Conclusion: A d-SPECT mismatch score of >or=4 was an appropriate cutoff at which diagnosis of myocardial ischemia in patients who were screened for ischemic heart disease. However, since patients with non-ischemic but organic heart disease can also present with abnormal mismatch findings, coronary angiography or CT might be warranted to differentiate IHD from non-IHD.

In the process of ovary sexual maturation, most immature ovarian follicles degrade into atretic follicles accompanied by apoptosis in granulosa cells. Macrophages can recognize apoptotic cells through specific binding with phosphatidylserine (PS), exposed on the surface of apoptotic cells, which is mediated by milk-fat globule-EGF factor 8 (MFG-E8). In the present research, we examined the involvement of the MFG-E8-dependent phagocytosis system in the atretic follicles of developing mouse ovaries. The number of atretic follicles and DNA-fragmented granulosa cells significantly increased in B6C3F1 mice during 2 to 6 weeks. Chromatin-condensed granulosa cells were engulfed by macrophages, which existed in the stroma or atretic follicles, or by neighboring normal granulosa cells. MFG-E8 mRNA increased in ovaries during 2 to 6 weeks, and immunoreactivity of MFG-E8 was detected at the surface of apoptotic cells existing around the antrum. Immunoelectron microscopic study revealed MFG-E8-positive signals on the membrane of apoptotic cells near macrophages, but apoptotic cells engulfed by neighboring granulosa cells showed few signals. Anti-Fas antibody elevated the annexin-V-positive reaction in isolated granulosa cells from 3-week-old mouse ovaries. MFG-E8 seems to act on the phagocytosis of apoptotic granulosa cells via macrophages and contribute to the regression process of atretic follicles.

Purpose: Diabetic patients with chronic kidney disease (CKD) frequently develop cardiac events within several years of the initiation of haemodialysis. The present study assesses the prognostic significance of stress myocardial ECG-gated perfusion imaging (MPI) in patients with diabetic CKD requiring haemodialysis., Methods: Fifty-five asymptomatic patients with diabetic stage V CKD and no history of heart disease scheduled to start haemodialysis were enrolled in this study (56 +/- 11 years old; 49 with type 2 diabetes mellitus). All patients underwent (201)Tl stress ECG-gated MPI 1 month before or after the initiation of haemodialysis to assess myocardial involvement. We evaluated SPECT images using 17-segment defect scores graded on a 5-point scale, summed stress score (SSS) and summed difference scores (SDS). The patients were followed up for at least 2 years (42 +/- 15 months) to determine coronary intervention (CI) and heart failure (HF) as soft events and acute myocardial infarction (AMI) and all causes of deaths as hard events., Results: The frequencies of myocardial ischaemia, resting perfusion defects, low ejection fraction and left ventricular (LV) dilatation were 24, 20, 29 and 49%, respectively. Ten events (18%) developed during the follow-up period including four CI, one HF, one AMI and four sudden deaths. Multivariate Cox analysis selected SDS (p = 0.0011) and haemoglobin A(1c) (HbA(1c)) (p = 0.0076) as independent prognostic indicators for all events., Conclusion: Myocardial ischaemia, in addition to glycaemic control, is a strong prognostic marker for asymptomatic patients with diabetic CKD who are scheduled to start haemodialysis. Stress MPI is highly recommended for the management and therapeutic stratification of such patients.

Temporal lobe epilepsy is a common form of pharmacoresistant epilepsy, in which epileptogenic foci propagate to other regions of the brain from the area of the initial insult. The present study focused on epileptogenesis, that is, the development of the first foci inducing seizures in amygdala-kindled mice, a model of temporal lobe epilepsy, to find the molecular process promoting the formation of epileptogenic networks. The expression of growth hormone (GH) was up-regulated along neural circuits during the epileptogenesis, while there was no difference in the pituitary gland. The up-regulation was associated with increased phosphorylation/activation of signal transducer and activator of transcription 5 and expression of the Serum Response Element-regulated genes, FBJ osteosarcoma oncogene, early growth response 1, and Jun-B oncogene, suggesting that expression of GH leads to GH signaling in the hippocampus and cortex. Furthermore, the administration of the hormone into the hippocampus markedly enhanced the progression of kindling. The administration of an inhibitor of its secretion into the hippocampus elicited a delay in the progression. Our results demonstrate directly that regulation via growth hormone has a robust impact in epileptogenesis.

Unlabelled: The quality of (18)F-FDG PET/CT images of overweight patients is often degraded. We evaluated the effect of optimizing injected dose or acquisition time on the quality of images of overweight patients using lutetium oxyorthosilicate PET/CT with high-performance detector electronics., Methods: We initially retrospectively measured radioactivity concentrations and signal-to-noise ratios (SNRs) in the liver relative to body weight for 80 patients who had undergone (18)F-FDG PET/CT according to our standard protocol (injected dose, 3.7 MBq/kg; acquisition time, 2 min/bed position). The patients were grouped (n = 20 per group) according to baseline body weight as G1 (or=85 kg). We compared the SNRs of G1 with those of G2, G3, and G4 and calculated the ratio squared as a factor to correct the acquisition parameters for overweight patients. We then prospectively enrolled 120 patients according to the same body weight criteria. We multiplied the correction factors to optimize injected doses or acquisition times and defined dose-adjusted groups (n = 20 per group) and time-adjusted groups (n = 20 per group). G2 dose was defined as 5.59 +/- 0.19 MBq/kg, G3 dose as 7.29 +/- 0.33 MBq/kg, and G4 dose as 8.88 +/- 0.43 MBq/kg. G2 time was defined as 3 min/bed position, G3 time as 4 min/bed position, and G4 time as 5 min/bed position., Results: Although liver activities did not significantly differ among G1 through G4 irrespective of patient weight, SNR progressively decreased as patient weight increased. The liver activities of G2 dose, G3 dose, and G4 dose were, respectively, 1.4-, 1.9-, and 2.5-fold higher than those of the baseline counterparts. Nevertheless, the increased liver activities of G2 dose, G3 dose, and G4 dose did not significantly affect SNR, compared with the baseline groups. In contrast, the SNR of G4 time was significantly higher than that of G4., Conclusion: Our findings suggest that the quality of images acquired from heavier patients can be maintained only by scanning for longer periods. Increasing the dose per kilogram of body weight did not improve the quality of lutetium oxyorthosilicate PET/CT images.

We found frequent and spontaneous proliferation of glandular epithelium and dilated cysts in the uteri of interleukin-2 receptor (IL2R) beta-chain transgenic (Tg2Rbeta) mice. The aim of this study was to examine the involvement of IL2R beta-chain in the pathogenesis of endometrial hyperplasia (EH). Mouse uteri and serum were collected from Tg2Rbeta and normal littermates (NL), which were classified into three groups according to age. The incidence of EH increased in an age-dependent manner in both types of mice. However, in old age, Tg2Rbeta mice showed more serious phenotypes of EH than NL. Immunohistochemical analysis revealed specific localization of IL2R beta-chain in the glandular epithelial cells, with a correlation to the degree of EH, not only in the Tg2Rbeta uteri but also in the NL uteri. Immunoreactions of CD3 and CD25 were detected in the uteri of Tg2Rbeta but were weak in the uteri of NL, and CD25-positive cells were distributed in the endometrial stroma and myometrium in the Tg2Rbeta mice. These findings suggest that the IL2R beta-chain induces growth potential for glandular epithelial cells and an immune-privileged condition mediated by CD25+regulatory-T cells.

To construct an optimization scheme for an extension of the Kohn-Sham approach, I introduce an operator form of the Coulomb interaction. This form is the sum of quadratic form pairs, which can be redefined in a self-consistent calculation of a multi-reference density functional theory. A detailed derivation of the form is given. A fluctuation term introduced in the extended Kohn-Sham scheme is expressed in this form for regularization. The present procedure also provides an exact derivation of effective negative interactions in charge fluctuation channels. Relevance to high-temperature superconductors is discussed.

The correct description for ion-radical systems has recently attracted much attention from density functional theory (DFT) researchers. Although several hybridization schemes using exact (Hartree-Fock) exchange and DFT exchange-correlation functionals have been proposed, it has been reported that such treatments do not work for the description of ion-radical systems. In this study we show that combining the exact exchange term in the Kohn-Sham DFT (or the Hartree-Fock equation) with the following resonating configuration interaction method is effective for the description of double-exchange type molecular magnetic interactions. The results are analyzed in relation to the 'many-electron self-interaction' concept that was recently proposed by DFT researchers.

Uterine natural killer (uNK) cells have roles for immune responses at the feto-maternal interface in mice. We studied the effects of beta(2)-microglobulin (beta(2)m) and perforin on proliferation and differentiation of uNK cells in pregnancy, using beta(2)-microglobulin-deficient (beta(2)m(-/-)) mice and perforin-deficient (P(-/-)) mice. The cell population of uNK cells in the metrial gland of P(-/-) mice was tended to be higher than the control B6 mice. The cell population of uNK cells in the metrial gland of beta(2)m(-/-) mice was significantly increased at Day 12 of pregnancy comparing to B6 and P(-/-) mice. On the other hand, the cell population of uNK cells in the decidua basalis of beta(2)m(-/-) mice was tended to be lower than B6 and P(-/-) mice. These results indicate that beta(2)m may be involved in proliferation of uNK cells in the metrial gland, and that beta(2)m may affect the maturation of uNK cells in the decidua basalis.

Objective: To evaluate the cytotoxicity of uterine natural killer (NK) cells and identify major biological factors fluctuating in the spontaneously aborted placenta., Design: Spontaneously aborted placentae were examined by using histology, molecular biology, and DNA microarray analysis., Setting: Laboratories and animal center at a medical college., Animal(s): C57BL/6 mice., Intervention(s): None., Main Outcome Measure(s): Up-regulation of the inhibitory system of uterine NK cells and enhancement of complement factors in aborted placenta., Result(s): In uteri on day 14 of pregnancy, 9.69% +/- 11.6% (+/-SD) implantation sites spontaneously aborted. Labyrinthine trophoblasts at abortion sites had nuclei that were positive for DNA fragment detection, as well as apoptotic morphology. Many uterine NK cells were present at abortion sites, but cytotoxic factors such as perforin, granzyme B, and Fas-Fas ligand were expressed at very low levels. In contrast, the expression level of Ly49 NK-cell receptors, which mediate an inhibitory signal for cytotoxicity, was enhanced at abortion sites. Deoxyribonucleic acid microarray analysis showed that adipsin, an activating enzyme for complement component C3, was strongly enhanced in aborted placenta, and positive reactions for adipsin and C3 were confirmed by immunohistochemistry., Conclusion(s): In the process of spontaneous abortion, the cytotoxicity of uterine NK cells is inhibited, and the innate immune system through adipsin and complement C3 appears to be influential.

This survey was performed in order to investigate the incidence of adverse reactions to radiopharmaceuticals in FY2006 in Japan. It was based on responses to questionnaires sent to nuclear medicine institutions. The reply was obtained from 975 institutions among 1263 to which the questionnaire had been sent. Thirty-two cases of adverse reactions were reported. A total of 1,189,127 radiopharmaceutical administrations was reported. The incidence of adverse reactions per 100,000 cases was 2.7. Three cases of defect products were reported, and the incidence of defect products per 100,000 cases was 0.3.

Lectin histochemistry was performed on mouse uteri to determine what effects leukemia inhibitory factor (LIF) has on carbohydrate epitope expressions at the time of implantation. Twenty-two biotinylated lectins were used in this study. Following injection of LIF, specific binding to the apical surface of the uterine glandular epithelium (GE) was recognized by six lectins. Particularly, binding of the lectin from Griffonia (Bandeiraea) simplicifolia was specific to the glandular epithelium close to the luminal epithelium. Succinylated wheat germ agglutinin (WGA), which has specificity for oligosaccharides recognized by WGA without sialic acid residues, showed weaker binding to the uterine luminal epithelium (LE) and the stroma than WGA, suggesting that terminal residues of glyco-conjugates on these tissues may be modified by sialic acids. Lectin binding to the glandular and luminal epithelium was not influenced by LIF. However, three lectins including a lectin from Dolichos biflorus showed specificity for stromal vessels 6h after LIF injection. Since the lectin from D. biflorus binds to neo-vascular vessels, LIF may play a role in regulating maternal angiogenesis directly and/or indirectly during implantation.

Objective: It has been reported that (123)I-metaiodobenzylguanidine (MIBG) scintigraphy can predict the poor prognosis in patients with dilated cardiomyopathy (DCM). However, the prognostic significance of MIBG is still unknown in patients with other heart diseases. In this study, we compared the prognosis and MIBG findings in various heart diseases., Methods: Consecutive 565 patients undergoing MIBG scintigraphy were enrolled (392 men, 52 +/- 16 years). Indications were that 127 had ischemic heart disease (IHD), 120 DCM, 101 hypertrophic cardiomyopathy (HCM), 21 hypertensive heart disease (HHD), 58 volume-load valvular disease (VVD), 38 pressure-load valvular disease (PVD), and 101 ventricular tachycardia or fibrillation (VTF). Heart-to-mediastinum ratio (H/M) and washout rate (WR) of MIBG were evaluated. Cardiac events were defined as sudden cardiac death, heart failure, and acute ischemic event (follow-up, 22.7 +/- 17.0 months)., Results: A total of 106 cardiac events including 40 cardiac deaths occurred. Cox hazard model analysis showed that in the IHD, HCM, and DCM groups, H/M and WR were associated with cardiac death, but not in the HHD, PVD, VVD, or VTF groups. Only death and congestive heart failure (CHF) episodes were related to H/M and WR. On the other hand, fatal arrhythmia, myocardial infarction, or angina pectoris were not related to H/M and WR. The data indicated that WR or H/M may predict death and CHF but does not predict fatal arrhythmia or acute ischemic event., Conclusions: MIBG WR and H/M were associated with heart failure, sudden death, and cardiac death events, and were useful to predict the prognosis in DCM, HCM, and IHD. In contrast, fatal arrhythmia events were not associated with MIBG indices, and thus it does not appear to be useful in predicting cardiac events in patients with VTF.

We have studied the ζ-phase of solid oxygen using the generalized gradient approximation in the density functional approach. Calculations of total energies and pressures have been carried out for the prototype of diatomic ζ-phase and other hypothetical monoatomic crystal structures. The diatomic phase was found to be stable over a wide range of pressure (100-2000 GPa). The stacking of molecular layers is discussed in comparison with the available experimental data.

It is shown that a momentum boost technique applied to the extended Kohn-Sham scheme enables the computational determination of the Mott insulating transition. Self-consistent solutions are given for correlated electron systems by first-principles calculations defined by the multi-reference density functional theory, in which the effective short-range interaction can be determined by the fluctuation reference method. An extension of the Harriman construction is made for the twisted boundary condition in order to define the momentum-boost technique in the first-principles manner. For an effectively half-filled-band system, the momentum-boost method tells that the period of a metallic ground state by the local density approximation (LDA) calculation is shortened to the least period of the insulating phase, indicating the occurrence of the Mott insulating transition.

Objective: (201)Thallium (TL), (99m)Tc-tetrofosmin (TF), and (99m)Tc-sestamibi (MIBI) are extensively used as myocardial perfusion agents. The objective of the present study was to evaluate their kinetics under acute ischemia-reperfusion., Methods: Isolated rat hearts, perfused by the Langendorff method at a constant flow rate of 10 ml/min, were allotted to normal control, mild ischemia, and severe ischemia groups, in which 20-min tracer wash-in was conducted followed by a 25-min tracer washout. No-flow ischemia (15 min for mild ischemia groups; 30 min for severe ischemia groups) was induced before conducting wash-in and washout in the ischemia groups. Whole-heart radioactivity was determined with an external gamma detector. Myocardial flow rate (K (1), ml/min) and clearance rate (k (2), min(-1)) were calculated., Results: K (1TL), K (1TF), and K (1MIBI) decreased according to the severity of ischemia (K (1TL) 5.32 +/- 0.53, 4.76 +/- 0.70, and 1.44 +/- 0.59; K (1TF) 3.80 +/- 0.70, 2.73 +/- 0.99, and 1.09 +/- 0.45; and K (1MIBI) 3.45 +/- 1.10, 2.15 +/- 0.82, and 1.05 +/- 0.13, in the normal control, mild, and severe ischemia groups, respectively). K (1) was significantly higher for TL than for the (99m)Tc tracers (P < 0.05), but the (99m)Tc tracers had equivalent K (1) values. k (2TL) increased significantly (P < 0.05) in the ischemia groups (k (2TL) 0.062 +/- 0.013, 0.11 +/- 0.045, and 0.12 +/- 0.035), but showed no significant difference between the ischemia groups. k (2MIBI) and k (2TF) were significantly (P < 0.05) lower than k (2TL) and increased significantly (P < 0.05) in the severe ischemia group (k (2TF) 0.0056 +/- 0.0022, 0.0037 +/- 0.0015, and 0.024 +/- 0.015; and k (2MIBI) 0.00072 +/- 0.0011, 0.00038 +/- 0.00076, and 0.042 +/- 0.034). k (2MIBI) was significantly (P < 0.05) lower than k (2TF) in the normal control and mild ischemia groups., Conclusions: Tracer extraction was higher for TL than for the (99m)Tc tracers and all tracers decreased according to the severity of ischemia-reperfusion in the three tracer groups. The clearance kinetics of not only MIBI but also TF is possibly useful for the evaluation of the severity of ischemia, and the Langendorff method and a methodological approach by continuous determinations of radioactivity may serve for the quantitative analysis of tracer kinetic profiles.

The effects of bisphenol A (BPA) on placentation have not been fully determined. The aim of this study was to clarify the structural changes of the placenta, abortion rate, and survival of neonates after BPA administration in mice. BPA (10 mg/kg/day) was administered to pregnant mice (BPA mice) subcutaneously from the first day of pregnancy (Day 0) to Day 7 (8 days total). The number of embryos and weights of whole uteri were measured on Days 10 and 12. Morphological changes in the placentae were examined by light microscopy on the corresponding days of pregnancy. The number of neonates was also counted. Survival rates were periodically calculated for neonates from the first day after parturition (P-Day 0) to P-Day 56. The number of embryos and weight of the uterus on Days 10 and 12 were significantly decreased by BPA injection. No notable differences were recognized between the left and right uteri. The proportion of the labyrinthine zone per whole placenta in the BPA mice became lower than that in the controls, and that of the metrial gland was higher in the BPA mice. The intervillous spaces of the placenta were narrower in the BPA mice. Degenerative changes were found in the trophoblastic giant cells and spongiotrophoblast layers of the BPA mice. The number of BPA mouse neonates was drastically decreased within 3 days after birth, and no mice survived after P-Day 56. The results suggest that BPA not only disrupts placental functions and leads to abortion through chronic stimulation of gene expression by binding to DNA but that it also affects the mortality of neonates through indirect exposure of embryos.

To clarify the mechanism of implantation, relationship between positioning of the mouse embryo in the uterus and distribution of uterine glands along the long axis of the uterine horn was examined by three-dimensional remodelling of the uterine endometrium. There were two unique regions in the endometrium. Uterine glands were distributed widely from mesometrial to anti-mesometrial side in one region. It was localized from lateral to anti-mesometrial side in another. These different regions were alternately aligned throughout the uterine horn. The number and position of embryos was consistent with that of the latter region. This study suggests that the type of distribution of uterine glands is closely related to the positioning of the embryo in mice.

Danazol, which has been used as a medicine for endometriosis, has a valid effect in pretreatment of patients receiving in vitro fertilization and embryo transfer, although its reproductive mechanism remains unclear. BALB/c mice were subcutaneously injected with danazol for 2 weeks. Blood and uteri were collected and cytokines were assayed. Following danazol treatment, an increase in pregnancy ratio was evident that was accompanied by up-regulation in serum macrophage-colony stimulating factor (M-CSF). RT-PCR analysis revealed that expression of M-CSF and Ly49, a phenotypic marker of natural killer (NK) cells, was up-regulated in the uteri of the danazol-treated mice. In immunohistochemical analysis, M-CSF and Ly49, together with alpha5 integrin, were clearly detected in the endometrium of the danazol-treated mice with very similar pattern of localization. These results suggest that danazol has an effect to promote pregnancy that induces recruitment of NK cells and a concomitant increase in the expression of M-CSF and alpha5 integrin in the uterus.

This survey was performed in order to investigate the incidence of adverse reactions to radiopharmaceuticals in FY2005 in Japan. It was based on responses to questionnaires sent to nuclear medicine institutions. The reply was obtained from 1,007 institutions among 1,243 to which the questionnaire had been sent. Nineteen cases of adverse reactions were reported. A total of 1,264,098 radiopharmaceutical administrations was reported. The incidence of adverse reactions per 100,000 cases was 1.5. Three cases of defect products were reported, and the incidence of defect products per 100,000 cases was 0.2.

We compared the reliability between apoptosis detection methods, namely, the terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) method and formamide-induced DNA denaturation assay using a monoclonal antibody (MAb) to single-stranded DNA (ssDNA) (formamide-MAb assay). Reaction targets in these methods are different: the TUNEL method recognizes free 3'-OH DNA ends, whereas the formamide-MAb assay detects ssDNA itself (25-30 bp). We found that the formamide-MAb assay immunohistochemically detected apoptotic cells, whereas the TUNEL method detected apoptotic cells as well as mitotic and necrotic cells. The TUNEL method recognized not only 3'-OH DNA ends cleaved by DNase during apoptosis but also constitutive physiological nicking that occurs in DNA duplication and histone posttranslational modifications during mitosis and random DNA breaks during necrotic execution. By electron microscopy, the mean labeling density (the number of 3'-OH DNA ends/nuclear area) obtained by the TUNEL method was determined to be consistently higher than that (the number of ssDNAs/nuclear area) obtained by the formamide-MAb assay. On the basis of these findings, we conclude that the formamide-MAb assay was more specific than the TUNEL method for the detection of apoptotic cells using electron microscopy; however, the labeling intensity of the formamide-MAb assay was slightly weaker than that of the TUNEL method.

We explore the unknown structure of phosphorus in phase IV (P-IV phase) based on first-principles calculations using the metadynamics simulation method. Starting from the simple cubic structure, we find a new modulated structure of the monoclinic lattice. The modulation is crucial to the stability of the structure. Through refining the structure further by changing the modulation period, we find the structure whose x-ray powder diffraction pattern is in best agreement with the experimental pattern. We expect that the modulation period of the structure in the P-IV phase is very close to that found in this study and probably incommensurate.

Background: Human breast cancer metastasizes mainly to lymph nodes, lungs, liver, and bone; in the majority of cases, it is the development of metastases which leads to death. In order to suppress mammary cancer metastasis, we applied in vivo electrogene transfer (non-viral method) as a means of interleukin-12 (IL-12) gene therapy on highly metastatic murine mammary cancer model., Methods: Metastatic mammary tumors induced by inoculation in BALB/c female mice were treated by intratumoral injections of either a plasmid vector containing IL-12 or empty vector and then subjected to in vivo electrogene transfer once a week for 8 weeks., Results: Treatment with IL-12 resulted in elevation of both IL-12 and IFNgamma levels in mammary tumors and in serum and intratumoral levels of CD4 and CD8 proteins were also increased. Tumor volumes and lymphatic and pulmonary metastases were significantly reduced. The histopathological changes induced by IL-12 characteristically included marked inflammation, increased apoptosis, decreased DNA synthesis, peripheral influx of significantly greater numbers of active macrophages, and reduced blood microvessel density, and apoptotic vascular endothelial cells were frequently seen. Western blotting showed decreases in VEGFR-3 of tumors exposed to IL-12 gene therapy. In adjuvant immunofluorescence studies, the CD31-positive endothelial cells of microvessels showed decreased VEGFR-3 expression in IL-12-treated tumors. However, apparent alterations in VEGFR-3 expression of podoplanin-positive lymphatic endothelial cells were not observed in IL-12-treated tumors. Although recombinant IL-12 did not inhibit tubular formation of human umbilical vein endothelial cells in a Matrigel assay, recombinant IFNgamma did completely suppress the tubular formation., Conclusions: In vivo electrogene transfer of IL-12 exerts strong anti-tumorigenic and anti-metastatic effects likely due to T-cell-mediated immune responses as well as anti-angiogenic action.

Unlabelled: Benign and malignant pulmonary lesions usually are differentiated by 18F-FDG PET with a semiquantitative 18F-FDG standardized uptake value (SUV) of 2.5. However, the frequency of malignancies with an SUV of <2.5 is significant, and pulmonary nodules with low 18F-FDG uptake often present diagnostic challenges., Methods: Among 360 consecutive patients who underwent 18F-FDG PET to evaluate pulmonary nodules found on CT, we retrospectively analyzed 43 who had solid pulmonary lesions (excluding lesions with ground-glass opacity, infiltration, or benign calcification) with an SUV of <2.5. The uptake of 18F-FDG was graded by a visual method (absent, faint, moderate, or intense) and 2 semiquantitative methods (SUV and contrast ratio [CR]). Final classification was based on histopathologic findings or at least 6 mo of clinical follow-up., Results: We found 16 malignant (diameter, 8-32 mm) and 27 benign (7-36 mm) lesions. When faint visual uptake was the cutoff for positive 18F-FDG PET results, the receiver-operating-characteristic (ROC) analysis correctly identified all 16 malignancies and yielded false-positive results for 10 of 27 benign lesions. Sensitivity was 100%, specificity was 63%, and the positive and negative predictive values were 62% and 100%, respectively. When an SUV of 1.59 was the cutoff for positive 18F-FDG PET results, the ROC analysis revealed 81% sensitivity, 85% specificity, and positive and negative predictive values of 77% and 89%, respectively. At a cutoff for positive 18F-FDG PET results of a CR of 0.29, the ROC analysis revealed 75% sensitivity, 82% specificity, and positive and negative predictive values of 71% and 85%, respectively. The areas under the curve in ROC analyses did not differ significantly among the 3 analyses (visual, 0.84; SUV, 0.81; and CR, 0.82). Analyses of intra- and interobserver variabilities indicated that visual and SUV analyses were quite reproducible, whereas CR analysis was poorly reproducible., Conclusion: These results suggested that for solid pulmonary lesions with low 18F-FDG uptake, semiquantitative approaches do not improve the accuracy of 18F-FDG PET over that obtained with visual analysis. Pulmonary lesions with visually absent uptake indicate that the probability of malignancies is very low. In contrast, the probability of malignancy in any visually evident lesion is about 60%.

This survey was performed in order to investigate the incidence of adverse reactions to radiopharmaceuticals and defect products in FY2004 in Japan. It was based on responses to questionnaires sent to nuclear medicine institutions. The reply was obtained from 968 institutions among 1,220 to which the questionnaire had been sent. Sixteen cases of adverse reactions were reported. A total of 1,277,906 radiopharmaceutical administrations was reported. The incidence of adverse reactions per 100,000 cases was 1.3. Eight cases of defect products were reported, and the incidence of defect products per 100,000 cases was 0.6.

We previously reported that all fetuses died or were resorbed on day 12 of pregnancy (Day 1= the day of plug) in interleukin-2 (IL-2) receptor beta-chain overexpressed transgenic (Tg2Rbeta) mice. In this study, to clarify the role of uterine natural killer (uNK) cells in pregnancy, the ultrastructure of Tg2Rbeta mouse uNK cells was analyzed using a transmission electron microscope. uNK cells and their granules on day 10 of pregnancy were larger in Tg2Rbeta mice than control mice, indicating that differentiation of uNK cells in Tg2Rbeta mice progressed rapidly. Additionally, the granules of uNK cells in Tg2Rbeta mice on day 10 of pregnancy had an irregular morphology. The multivesicular regions were present in the cap structure of these granules, suggesting that the uNK cells of the Tg2Rbeta mice had cytotoxic activity.

Objective: The feasibility of human cardiac imaging using a prototype 256-detector row cone-beam computed tomography (256CBCT) scanner without electrocardiographic gating was examined., Methods: Two healthy male volunteers were examined by contrast-enhanced 256CBCT. The number of detectors was 912 x 256, each measuring approximately 0.5 mm x 0.5 mm at the center of rotation. The craniocaudal coverage was approximately 100 mm after reconstruction by the Feldkamp-Davis-Kress algorithm. The effective time resolution was 500 milliseconds using a half-scan algorithm., Results: Serial enhancement of the left ventricular myocardium was detected. The right and left coronary arteries at proximal and distal segments were depicted without significant blurring. Although the left ventricular wall motion on cine images was not smooth over time, it was possible to measure ventricular volume and ejection fraction., Conclusions: Using the 256CBCT, it was possible to visualize the coronary arteries, myocardial perfusion, and ventricular contraction simultaneously during a single acquisition.

Uterine natural killer (uNK) cells in the pregnant uterus are known to be associated with the normal development of the placenta. In the mouse pregnant uterus, many uNK cells exist during mid pregnancy, although they show a sudden decrease during late pregnancy and almost disappear before delivery. Our previous study indicated that uNK cells showed clear apoptotic morphology during late pregnancy. Therefore, the present study was carried out to define the involvement of Fas ligand (FasL) and Fas in apoptosis induction of uNK cells. Immunohistochemical analyses revealed that uNK cells expressed FasL in the cytoplasmic granules and Fas on the cell membrane during late pregnancy. In lpr/lpr mice, which genetically lack Fas, many uNK cells were clearly observed during late pregnancy compared with wild-type mice, and moreover uNK cells still existed at day-18 of pregnancy, although there were few in wild-type mice during the same period. In the experiment of in vitro culture, uNK cells derived from wild-type placenta showed chromatin condensation and DNA fragmentation frequently following the anti-Fas antibody treatment, as compared with the control. From these results, it is suggested that FasL and Fas-dependent apoptosis regulates cell appearance of uNK cells in the mouse pregnant uterus.

Radio-iodine (131I) therapy has been using in Graves' disease and well differentiated thyroid cancer. The rules of control in the discharge from radio-isotope hospital were notified in 1999 in Japan. Guideline of the 131I therapy in Graves' disease and thyroid cancer were prepared by sub-group of Japanese Society of Nuclear Medicine.

In recent years, positron-emitting labelled radiopharmaceuticals have come to be used in conjunction with positron emission tomography (PET) in various clinical diagnoses. Radiation exposure of the medical staff is a key issue in the design of PET facilities intended to handle large numbers of persons for PET diagnosis. As a first step, the radiation dose to individuals who received radiopharmaceuticals was calculated using a mathematical phantom model and the EGS4 electromagnetic cascade Monte Carlo code and MCNP Monte Carlo code. Dose rate behind a lead shield was also calculated for various lead thicknesses. The radiation dose distribution around a syringe containing a positron emitter was calculated. The calculated dose distributions were fitted to polynomial equations. These calculations were evaluated against measurements. The second step was to evaluate medical staff dose at a specified time by superimposing dose distribution from each person who received radioisotopes taking into account radioactive decay. In this way, we developed software to support PET facility operation, namely, planning, prediction, control of medical staff dose and facility operation. This system was also designed to schedule daily radiopharmaceuticals production and to manage radioactive wastes by taking decay time into account.

Background: Although sentinel lymph node biopsy(SLNB)is highly accurate in predicting axillary nodal status in patients with breast cancer, it has been shown that the procedure is associated with a few false negative results. The risk of leaving metastatic nodes behind in the axillary basin when SLNB is negative should be estimated for an individual patient if SLNB is performed to avoid conventional axillary lymph node dissection(ALND)., Methods: A retrospective analysis of 512 women with T1-3N0M0 breast cancer was conducted to derive a prevalence of nodal metastasis by T category as a pre-test(i.e., before SLNB)probability and to examine potential confounders on the relationship between T category and axillary nodal involvement. Probability of nodal metastasis when SLNB was negative was estimated by means of Bayes' theorem which incorporated the pre-test probability and sensitivity and specificity of SLNB., Results: Axillary nodal metastasis was observed in 6.1% of T1a-b, 25.1% of T1c, 28.7% of T2, 35.0% of T3 tumors. Point estimates for the probability of nodal involvement when SLNB was negative ranged from 0.3-1.3% for T1a-b, 1.6-6.3% for T1c, 2.0-7.5% for T2, and 2.6-9.7% for T3 tumors with representative sensitivities of 80%, 85%, 90% and 95%, respectively. The risk may be higher when the tumor involves the upper outer quadrant of the breast, while it may be lower for an underweight woman., Conclusions: The probability of axillary lymph node metastasis when SLNB is negative can be estimated using a Bayesian approach. Presenting the probability to the patient may guide the decision of surgery without conventional ALND.

The pore-forming protein (perforin) produced by lymphocytes can induce apoptosis in target cells. In mouse placenta, although a large amount of perforin is produced by the uterine natural killer (uNK) cells, its role in the reproductive process is still not clear. Since the cytotoxicity of uNK cells can be enhanced by interleukin (IL)-2, we studied the role of perforin in the placenta of wild-type and perforin-knockout mice treated with IL-2 during days 10-14 of pregnancy. Immunohistochemistry of the wild-type mice showed that the perforin was positive in the membrane of trophoblast glycogen cells as well as the cytoplasm of uNK cells, and there was an increase in the expression level following IL-2 treatment as revealed by RT-PCR analysis, although no change was identified in fertility. In the IL-2-treated perforin-knockout mice, however, the number of live fetuses was decreased, accompanied by an increase in the weight of placentae. Examination of these placentae showed an abnormally enlarged junctional zone, occupied by a large number of the trophoblast glycogen cells and significantly few of the apoptotic cells. These findings indicate that perforin can contribute to a successful pregnancy by inhibiting the excessive growth of the junctional zone induced by IL-2.

We investigated the effectiveness of in vivo electrogene transfer as a means of therapy in rat urinary bladder carcinoma and in mammary carcinoma models in both athymic and syngeneic mice using the herpes simplex virus 1 thymidine kinase (HSVtk) or IL-12 genes in combination with ganciclovir (GCV). A significant increase in the levels of tissue apoptosis and necrosis was induced with a single injection of HSVtk vector directly into bladder and mammary tumors followed by in vivo transfection and a regimen of intraperitoneal GCV injection. This procedure induced significant selective tumor cell death, characterized by marked inflammation and peripheral macrophage influx. Active caspase-3 was also strongly expressed in areas of cell death, indicating the initiation of apoptosis. This result was confirmed in corollary in vitro studies on a mouse bladder carcinoma cell line in which elevated caspase-3, -8, and -9 activities and decreased mitochondrial membrane potential were observed as a result of transfection with HSVtk and addition of GCV to the medium. In the syngeneic mouse mammary cancer model, we additionally found both tumor volume and metastasis to lymph nodes and lungs to be significantly reduced throughout the 2-month experiment. However, in contrast to their syngeneic counterparts, HSVtk/GCV therapy did not effectively inhibit mammary tumor growth/metastasis in an athymic mouse model, leading us to believe that T-cell-mediated immune responses may participate via the bystander effect in HSVtk/GCV experimental therapy. We subsequently evaluated the antitumor activity of IL-12, which can activate T-cell-mediated immune responses involving macrophages, in the syngeneic mammary tumors and found that IL-12 also significantly suppressed mammary tumor growth and metastasis. We thus suggest that in vivo electrogene transfer is a useful transfection tool in cancer gene therapy and, in addition, we show that T-cell-mediated immune responses may be a critical factor in cancer gene therapy using HSVtk/GCV and IL-12.

To explore the possibility of which medical radioactive wastes could be disposed as general wastes after keeping them a certain period of time and confirming that their radioactivity reach a background level (BGL), we made a survey of these wastes in several nuclear medicine facilities. The radioactive wastes were collected for one week, packed in a box according to its half-life, and measured its radioactivity by scintillation survey meter with time. Some wastes could reach a BGL within 10 times of half-life, but 19% of the short half-life group (group 1) including 99mTc and 123I, and 8% of the middle half-life group (group 2) including 67Ga, (111)In, and 201Tl did not reach a BGL within 20 times of half-life. A reason for delaying the time of reaching a BGL might be partially attributed to high initial radiation dose rate or heavy package weight. However, mixing with the nuclides of longer half-life was estimated to be the biggest factor affecting this result. When disposing medical radioactive wastes as general wastes, it is necessary to avoid mixing with radionuclide of longer half-life and confirm that it reaches a BGL by actual measurement.

We conducted a questionnaire survey about radiation-safety management condition in Japanese nuclear medicine facilities to make materials of proposition for more reasonable management of medical radioactive waste. We distributed a questionnaire to institutions equipped with Nuclear Medicine facilities. Of 1,125 institutions, 642 institutes (52.8%) returned effective answers. The questionnaire covered the following areas: 1) scale of an institution, 2) presence of enforcement of radiotherapy, 3) system of a tank, 4) size and number of each tank, 5) a form of draining-water system, 6) a displacement in a radioactive rays management area, 7) a measurement method of the concentration of medical radioactive waste in draining water system, 8) planned and used quantity of radioisotopes for medical examination and treatment, 9) an average displacement of hospital for one month. In most institutions, a ratio of dose limitation of radioisotope in draining-water system was less than 1.0, defined as an upper limitation in ordinance. In 499 hospitals without facilities of hospitalization for unsealed radioisotope therapy, 473 hospitals reported that sum of ratios of dose limits in a draining-water system was less than 1.0. It was calculated by used dose of radioisotope and monthly displacement from hospital, on the premise that all used radioisotope entered in the general draining-water system. When a drainage including radioactivity from a controlled area join with that from other area before it flows out of a institution, it may be diluted and its radioactive concentration should be less than its upper limitation defined in the rule. Especially, in all institutions with a monthly displacement of more than 25,000 m3, the sum of ratio of the concentration of each radionuclide to the concentration limit dose calculated by used dose of radioisotope, indicated less than 1.0.

A survey on the I-131 therapy of Graves' hyperthyroidism was undertaken by questionnaire in 1,246 hospitals of Japan. One thousand and ninety seven of them (88.0%) responded to the questionnaire. In this paper, we report the results and analysis of the replies to the questionnaire. In the 121 hospitals (11.03%) of the respondents, I-131 therapy is being performed for Graves' hyperthyroidism. A gradual increase was observed in the annual number of I-131 treated Graves' disease patients during the period of 1998-2001, from 1,740 to 2,484. I-131 treatment was selected mainly for the cases with side effects from antithyroid drug (ATD) therapy, followed by the cases with complication of heart or hepatic diseases, recurrences of hyperthyroidism after surgery, radioiodine treatment, and long-term ATD treatment. The 41% of respondents used I-131 in order to restore euthyroidism, 34% aimed for hyperthyroidism and 41% used the dose properly between the two according to the patients. Administration dosage of I-131 was estimated mainly on the basis of thyroid uptake and volume in 93% of the respondents and 48% calculated the radiation dose by also determining the effective half-life in the thyroid gland. Thyroid size was estimated by scintigram (51%), US (33%), CT (22%) and palpation (12%). ATD treatment was used before I-131 administration by 70% of the respondents and 34% after radioiodine therapy. A low-iodine diet was given to the patients for a week (46%) or two weeks (47%) before I-131 administration. However, after treatment only 46% of the respondents continued low-iodine diet for a week.

Localization of apoptotic cells in the kidney of perinatal rats was examined by the terminal deoxynucleotidyl transferase-mediated d-UTP-biotin nick end labeling (TUNEL) method and electron microscopy. Perinatal changes in the percentage of kidney cells with DNA fragmentation were determined by flow cytometric analysis. Through observation of two successive sections, the relationship between the localization of the epidermal growth factor receptor (EGFR) positive cells and TUNEL positive cells in the kidney was determined. From fetal day 18 to neonatal day 5, TUNEL positive cells were noted in immature glomeruli, collecting ducts and interstitium. Electron microscopically, chromatin condensed nuclei and apoptotic bodies were seen in the same tissue component as the TUNEL positive cells. The percentage of DNA fragmented cells significantly increased from fetal days 18 to 20 and significantly decreased from fetal days 20 to 22, while they still remained low in the neonatal period. The TUNEL positive cells in immature glomeruli and collecting ducts were not reactive to the EGFR antibody. The TUNEL positive cells were not observed in the proximal tubular cells, which were positive to EGFR antibody. These results indicate that apoptotic cells are present in the kidney throughout the perinatal period in the rat and that EGF plays an important role in perinatal development of the rat kidney.