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To detect and track structural changes in atomic nuclei, the systematic study of nuclear levels with firm spin-parity assignments is important. While linear polarization measurements have been applied to determine the electromagnetic character of gamma-ray transitions, the applicable range is strongly limited due to the low efficiency of the detection system. The multi-layer Cadmium-Telluride (CdTe) Compton camera can be a state-of-the-art gamma-ray polarimeter for nuclear spectroscopy with the high position sensitivity and the detection efficiency. We demonstrated the capability to operate this detector as a reliable gamma-ray polarimeter by using polarized 847-keV gamma rays produced by the 56 Fe ( p , p ′ γ ) reaction. By combining the experimental data and simulated calculations, the modulation curve for the gamma ray was successfully obtained. A remarkably high polarization sensitivity was achieved, compatible with a reasonable detection efficiency. Based on the obtained results, a possible future gamma-ray polarimetery is discussed. [ABSTRACT FROM AUTHOR]

FOXA1 expression correlates with the breast cancer luminal subtype and patient survival. RNA and protein analysis of a panel of breast cancer cell lines revealed that BRCA1 deficiency is associated with the downregulation of FOXA1 expression. Knockdown of BRCA1 resulted in the downregulation of FOXA1 expression and enhancement of FOXA1 promoter methylation in MCF-7 breast cancer cells, whereas the reconstitution of BRCA1 in Brca1-deficent mouse mammary epithelial cells (MMECs) promoted Foxa1 expression and methylation. These data suggest that BRCA1 suppresses FOXA1 hypermethylation and silencing. Consistently, the treatment of MMECs with the DNA methylation inhibitor 5-aza-2′-deoxycitydine induced Foxa1 mRNA expression. Furthermore, treatment with GSK126, an inhibitor of EZH2 methyltransferase activity, induced FOXA1 expression in BRCA1-deficient but not in BRCA1-reconstituted MMECs. Likewise, the depletion of EZH2 by small interfering RNA enhanced FOXA1 mRNA expression. Chromatin immunoprecipitation (ChIP) analysis demonstrated that BRCA1, EZH2, DNA methyltransferases (DNMT)1/3a/3b and H3K27me3 are recruited to the endogenous FOXA1 promoter, further supporting the hypothesis that these proteins interact to modulate FOXA1 methylation and repression. Further co-immunoprecipitation and ChIP analysis showed that both BRCA1 and DNMT3b form complexes with EZH2 but not with each other, consistent with the notion that BRCA1 binds to EZH2 and negatively regulates its methyltransferase activity. We also found that EZH2 promotes and BRCA1 impairs the deposit of the gene silencing histone mark H3K27me3 on the FOXA1 promoter. These associations were validated in a familial breast cancer patient cohort. Integrated analysis of the global gene methylation and expression profiles of a set of 33 familial breast tumours revealed that FOXA1 promoter methylation is inversely correlated with the transcriptional expression of FOXA1 and that BRCA1 mutation breast cancer is significantly associated with FOXA1 methylation and downregulation of FOXA1 expression, providing physiological evidence to our findings that FOXA1 expression is regulated by methylation and chromatin silencing and that BRCA1 maintains FOXA1 expression through suppressing FOXA1 gene methylation in breast cancer. [ABSTRACT FROM AUTHOR]

The E2 ubiquitin conjugating enzyme Ubc13 and the E3 ubiquitin ligases Rad18 and Rnf8 promote homologous recombination (HR)-mediated double-strand break (DSB) repair by enhancing polymerization of the Rad51 recombinase at γ-ray-induced DSB sites. To analyze functional interactions between the three enzymes, we created RAD18−/−, RNF8−/−, RAD18−/−/RNF8−/− and UBC13−/−clones in chicken DT40 cells. To assess the capability of HR, we measured the cellular sensitivity to camptothecin (topoisomerase I poison) and olaparib (poly(ADP ribose)polymerase inhibitor) because these chemotherapeutic agents induce DSBs during DNA replication, which are repaired exclusively by HR. RAD18−/−, RNF8−/− and RAD18−/−/RNF8−/− clones showed very similar levels of hypersensitivity, indicating that Rad18 and Rnf8 operate in the same pathway in the promotion of HR. Although these three mutants show less prominent defects in the formation of Rad51 foci than UBC13−/−cells, they are more sensitive to camptothecin and olaparib than UBC13−/−cells. Thus, Rad18 and Rnf8 promote HR-dependent repair in a manner distinct from Ubc13. Remarkably, deletion of Ku70, a protein essential for nonhomologous end joining (NHEJ) significantly restored tolerance of RAD18−/− and RNF8−/− cells to camptothecin and olaparib without affecting Rad51 focus formation. Thus, in cellular tolerance to the chemotherapeutic agents, the two enzymes collaboratively promote DSB repair by HR by suppressing the toxic effect of NHEJ on HR rather than enhancing Rad51 focus formation. In contrast, following exposure to γ-rays, RAD18−/−, RNF8−/−, RAD18−/−/RNF8−/− and UBC13−/−cells showed close correlation between cellular survival and Rad51 focus formation at DSB sites. In summary, the current study reveals that Rad18 and Rnf8 facilitate HR by two distinct mechanisms: suppression of the toxic effect of NHEJ on HR during DNA replication and the promotion of Rad51 focus formation at radiotherapy-induced DSB sites. [ABSTRACT FROM AUTHOR]

Human T-cell leukemia virus type 1 (HTLV-1), which causes adult T-cell leukemia (ATL) in humans, establishes a life-long latent infection. Current therapies are not very effective against HTLV-1-associated disorders. A novel therapeutic approach may help to combat HTLV-1 infection. A molecular therapy that targets the proviral genome is favorable because the therapeutic effect occurs specifically in HTLV-1-infected cells, regardless of whether they express viral genes. In this proof-of-concept study, we developed a therapeutic molecule based on zinc finger nuclease (ZFN) to achieve this goal. We designed a ZFN that specifically recognized conserved region of HTLV-1 long terminal repeat (LTR) and introduced it into various HTLV-1-positive human T-cell lines, including HTLV-1-transformed and ATL-derived cell lines. The ZFN disrupted the promoter function of HTLV-1 LTR and specifically killed HTLV-1-infected cells. We also showed a potential approach of this therapeutic molecule to remove the proviral genome from HTLV-1-infected cells, something that has not been possible before. The therapeutic effect of ZFN was confirmed in an in vivo model of ATL. This strategy may form the basis of a therapy that can eradicate HTLV-1 infection. Similar approaches can be used to target other malignancy-associated viruses. [ABSTRACT FROM AUTHOR]

The Pulse Shape Processor is a digital signal processing electronics for the microcalorimeter instrument onboard ASTRO-H. Receiving digitized waveform (14 bit, 12.5 kHz sample) from 2×18 channels, two identical units of PSP-A and -B trigger X-ray events, assign five kinds of event grade, and perform optimal filtering to measure energy deposit on the 6×6 microcalorimeter pixels. One unit of PSP is composed of one FPGA board and two CPU boards. This paper describes the event processing algorithm to fulfill requirements for the signal processing, and task sharing between FPGA and CPU. [ABSTRACT FROM AUTHOR]

Hemoglobin (Hb) Hiroshima is an Hb variant that travels rapidly on electrophoresis and shows a fourfold increase in oxygen affinity and a decreased Bohr effect. We encountered a 40-year-old male patient with erythremia and an undetectable HbA(1c) level. The presence of an abnormal hemoglobin molecule was suggested by the results of high-performance liquid chromatography analysis. Subsequent gene analysis by direct sequencing confirmed Hb Hiroshima (β146 histidine → aspartic acid). Caution should be exercised when diagnosing erythremia. [ABSTRACT FROM AUTHOR]

Although several studies have reported that locally administering oncolytic viruses effectively targets malignancies, the efficacy of systemically administered oncolytic viruses is restricted. Recently, however, it was reported that systemic administration of oncolytic vesicular stomatitis virus adsorbed onto antigen-specific lymphocytes was effective against malignancies. We hypothesized that intravenously administering such virus might have significant potential in treatment of the malignant tumors. We adsorbed oncolytic herpes simplex virus-1 mutant R3616 onto lymphocytes harvested from mice with acquired antitumor immunity. We administered adsorbed R3616 to peritoneally disseminated tumors and analyzed the efficacy of this treatment. Mice administered adsorbed R3616 survived significantly longer than mice administered R3616 adsorbed onto non-specific lymphocytes, or mice administered either virus or tumor antigen-specific lymphocytes alone. In this context, herpes oncolytic virus is a promising treatment not only for primary lesions, but also for multiple metastasizing lesions. This treatment strategy may become one of the most effective methods for systemic virus delivery. [ABSTRACT FROM AUTHOR]

Oncolytic viruses are a promising method of cancer therapy, even for advanced malignancies. HF10, a spontaneously mutated herpes simplex type 1, is a potent oncolytic agent. The interaction of oncolytic herpes viruses with the tumor microenvironment has not been well characterized. We injected HF10 into tumors of patients with recurrent breast carcinoma, and sought to determine its effects on the tumor microenvironment. Six patients with recurrent breast cancer were recruited to the study. Tumors were divided into two groups: saline-injected (control) and HF10-injected (treatment). We investigated several parameters including neovascularization (CD31) and tumor lymphocyte infiltration (CD8, CD4), determined by immunohistochemistry, and apoptosis, determined by terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Median apoptotic cell count was lower in the treatment group (P=0.016). Angiogenesis was significantly higher in treatment group (P=0.032). Count of CD8-positive lymphocytes infiltrating the tumors was higher in the treatment group (P=0.008). We were unable to determine CD4-positive lymphocyte infiltration. An effective oncolytic viral agent must replicate efficiently in tumor cells, leading to higher viral counts, in order to aid viral penetration. HF10 seems to meet this criterion; furthermore, it induces potent antitumor immunity. The increase in angiogenesis may be due to either viral replication or the inflammatory response. [ABSTRACT FROM AUTHOR]

Transition process of optical modal properties induced by the introduction of randomness into random photonic crystals is investigated by a computational method. We analyze an impulse response of two-dimensional triangular photonic crystals, in which the positions of the air holes are slightly deviated to random directions. It is shown that the appropriate degree of random departure from a perfect crystal state gives rise to multiple scattering of low group velocity band-edge modes and supports their strong Anderson localization. The achieved confinement efficiency of light exceeds the one obtained in the perfect photonic crystal state. [ABSTRACT FROM AUTHOR]

Duchenne muscular dystrophy (DMD) is the most common form of the progressive muscular dystrophies characterized by defects of the dystrophin gene. Although primarily characterized by degeneration of the limb muscles, cardiomyopathy is a major cause of death. Therefore, the development of curative modalities such as gene therapy is imperative. We evaluated the cardiomyopathic features of mdx mice to observe improvements in response to intravenous administration of recombinant adeno-associated virus (AAV) type 9 encoding microdystrophin. The myocardium was extensively transduced with microdystrophin to significantly prevent the development of fibrosis, and expression persisted for the duration of the study. Intraventricular conduction patterns, such as the QRS complex duration and S/R ratio in electrocardiography, were also corrected, indicating that the transduced microdystrophin has a protective effect on the dystrophin-deficient myocardium. Furthermore, BNP and ANP levels were reduced to normal, suggesting the absence of cardiac dysfunction. In aged mice, prevention of ectopic beats as well as echocardiographic amelioration was also demonstrated with improved exercise performance. These findings indicate that AAV-mediated cardiac transduction with microdystrophin might be a promising therapeutic strategy for the treatment of dystrophin-deficient cardiomyopathy. [ABSTRACT FROM AUTHOR]

In 2005, we initiated a clinical trial that examined the efficacy of the oncolytic virus HF10 to treat pancreatic cancer. Pancreatic cancer continues to have a high mortality rate, despite multimodal treatments for patients, and new therapeutic methods are greatly needed. The current mainstream methods for cancer treatment include biological therapeutics such as trastuzumab (Herceptin) for breast cancer or erlotinib (Tarceva) for non-small cell lung cancer. Oncolytic virus therapy is a new and promising treatment strategy for cancer. Oncolytic viruses are novel biological therapeutics for advanced cancer that appear to have a wide spectrum of anticancer activity with minimal human toxicity. To examine the efficacy of oncolytic virus therapy for pancreatic cancer, we initiated pilot studies by injecting six patients with non-resectable pancreatic cancer with three doses of HF10. All patients were monitored for 30 days for local and systemic adverse effects and were not administered any other therapeutics during this period. There were no adverse side-effects, and we observed some therapeutic potential based on tumor marker levels, survival, pathological findings and diagnostic radiography. The tumors were classified as stable disease in three patients, partial response in one patient and progressive disease in two patients. [ABSTRACT FROM AUTHOR]

The estrogen-responsive gene Efp promotes the growth of breast cancer cells by stimulating the degradation of a negative cell-cycle regulator, 14-3-3σ, and is hence considered a suitable molecular target for breast cancer therapy. The use of small interfering RNA (siRNA) and its derivatives to silence cancer-related genes is being investigated with the aim of identifying clinical applications for these molecules. Recently, it has been shown that DNA-modified siRNA (chimeric siRNA) has good potential in clinical applications, because it induces fewer off-target effects or immune responses in mammalian cells. In the present study, we identified the most specific and effective siRNA (siEfp-1) for silencing Efp expression in MCF-7 breast cancer cells. For this purpose, we used an algorithm that primarily eliminates off-target effects. siEfp-1 considerably suppressed the in vitro proliferation and cell-cycle progression of MCF-7 cells, as well as the in vivo growth of MCF-7 tumors, in athymic mice. DNA-modified siEfp-1 (chimeric siEfp) significantly inhibited the expression of Efp, proliferation of cultured cells and the in vivo growth of MCF-7-derived tumors in athymic mice. In addition, the silencing of Efp expression by siEfp-1 and chimeric siEfp increased the expression of the 14-3-3σ protein. These results suggest that siEfp-1 and chimeric siEfp could be useful in breast cancer therapy. Chimeric siEfp, in particular, has a high specificity and induces few side effects and is therefore expected to be used as a novel nucleic acid-based therapeutic agent. [ABSTRACT FROM AUTHOR]

The modal characteristics of localized electromagnetic waves inside random nanostructures are theoretically presented. It is crucial to know the tuneabilities of the localized modes systematically for demonstrating a specific random lasing application. By use of FDTD (Finite-Difference Time-Domain) method, we investigated the impulse response of two-dimensional random nanostructures consisting of closely packed cylindrical dielectric columns, and precisely analyzed the localized modes. We revealed the tuneability of the frequency of the localized modes by controlling the medium configurations: diameter, spatial density, and refractive index of the cylinders. Furthermore, it is found to be able to tune the Q (quality) factors of the localized modes dramatically by controlling simply the system size of the entire medium. The observed Q factors of approximately 1.6×104 were exhibited in our random disordered structures. [ABSTRACT FROM AUTHOR]

Abnormal centrosome numbers arise in tumours and can cause multipolar mitoses and genome instability. Cdk2 controls normal centrosome duplication, but Chk1-dependent centrosome amplification also occurs after DNA damage. We investigated the involvement of cyclin-dependent kinases (Cdks) in DNA damage-induced centrosome amplification using cells lacking either Cdk2, or both Cdk1 and Cdk2 activity. Cdk2−/− DT40 cells showed robust centrosome amplification after ionizing radiation (IR), whereas Cdk1-deficient Cdk2−/− cells showed no centrosome amplification, demonstrating that Cdk1 can substitute for Cdk2 in this pathway. Surprisingly, we found that Cdk2 activity was upregulated by IR in wild-type but not in Chk1−/− DT40 cells. Cdk2 upregulation also occurred in HeLa cells after IR treatment. Chk1-dependent Cdk2 induction was not accompanied by increased levels of Cdk1, Cdk2, cyclin A or cyclin E, but activating T160 phosphorylation of Cdk2 increased after IR. Moreover, Cdk2 overexpression restored IR-induced centrosome amplification in Cdk1-deficient Cdk2−/− cells, but T160A mutation blocked this rescue. Our data suggest that Chk1 signalling causes centrosome amplification after IR by upregulating Cdk2 activity through activating phosphorylation. [ABSTRACT FROM AUTHOR]

Measurement of brachial-ankle pulse wave velocity (baPWV) is recognized as a simple and practical method for assessing arterial stiffness. We determined whether the baPWV of adolescents is affected by obesity and its associated metabolic risk variables. A cross-sectional sample of 754 apparently healthy adolescents (383 men and 371 women), aged 15–17 years, was recruited for this study. baPWV was measured by a simple automatic oscillometric technique. Adiposity measures, blood pressure, serum lipoproteins, fasting glucose and insulin were evaluated. The baPWV of the adolescents was significantly higher in men than in women and increased with age in both genders. After being statistically adjusted for age and gender, baPWV was significantly correlated with body mass index, percent body fat, waist-to-height ratio, systolic and diastolic blood pressures, mean arterial pressure, triglycerides, high-density lipoprotein cholesterol (HDL-C), atherogenic index, glucose, insulin, and homoeostasis model assessment of insulin resistance (HOMA-IR). In the multivariate regression analysis, mean arterial pressure, atherogenic index, HOMA-IR, systolic blood pressure and age were found to be significant determinants of baPWV (P<0.001). An increasing number of clustered risk variables, including high values (>gender-specific top quartiles) of waist-to-height ratio, mean arterial pressure, atherogenic index and HOMA-IR showed a graded association with baPWV (P<0.001 for trend). These results suggest that obesity and its associated metabolic abnormalities are important factors in the increased baPWV of adolescents and that baPWV may be useful in investigating early arterial wall changes in this population.Journal of Human Hypertension (2009) 23, 444–450; doi:10.1038/jhh.2008.143; published online 11 December 2008 [ABSTRACT FROM AUTHOR]

We report on a new physical aspect of random lasing, an extremely selective modal oscillation by using a two-dimensional calculation model. The developed model consists of two theoretical subsystems, two-dimensional scattering model for describing multiple scattering process and rate equation model for describing lasing process. Using this model, we show how emission spectrum behaves as the scattering state of photons inside the system changes. It is shown that specific and strong modal oscillation takes place in a closed loop path of emitted photons, efficiently supported by the background multiple scattering. With the increase of multiple scattering events around the closed loop path, the system starts to oscillate with an extremely strong mode without any ASE (amplified spontaneous emission) noise. [ABSTRACT FROM AUTHOR]

Microbeam analysis is used in biomedical and environmental sciences to determine the presence and concentration of trace elements. However, quantitative analyses of biological samples are challenging because the chemical and physical compositions of existing standards differ from those typically encountered in biological samples. We developed a thin standard using polyvinyl alcohol and assessed its quality by microbeam scanning particle induced X-ray emission (micro-PIXE) analysis. The relationship between metal concentration and X-ray intensity was linear for certain standards up to 500 μg·g−1. Using this new thin standard, micro-PIXE analysis of Zn content in samples of human hair agreed well with analysis performed by inductively-coupled plasma mass spectrometry, validating the use of these new thin standards for quantitative mapping with microbeam analysis. [ABSTRACT FROM AUTHOR]

Alteration in transforming growth factor-beta signalling pathway is one of the main causes of pancreatic cancer. The human runt-related transcription factor 3 gene (RUNX3) is an important component of this pathway. RUNX3 locus 1p36 is commonly deleted in a variety of human cancers, including pancreatic cancer. Therefore, we examined genetic and epigenetic alterations of RUNX3 in human pancreatic cancer. Thirty-two patients with pancreatic cancer were investigated in this study. We examined the methylation status of RUNX3 promoter region, loss of heterozygosity (LOH) at 1p36, and conducted a mutation analysis. The results were compared with clinicopathological data. Promoter hypermethylation was detected in 20 (62.5%) of 32 pancreatic cancer tissues, confirmed by sequence of bisulphite-treated DNA. Loss of heterozygosity was detected in 11 (34.3%) of 32 pancreatic cancers. In comparison with clinicopathological data, hypermethylation showed a relation with a worse prognosis (P=0.0143). Hypermethylation and LOH appear to be common mechanisms for inactivation of RUNX3 in pancreatic cancer. Therefore, RUNX3 may be an important tumour suppressor gene related to pancreatic cancer. [ABSTRACT FROM AUTHOR]

A new therapeutic approach to treat Alzheimer's disease (AD) is needed, and the use of growth factors is considered to be a candidate. Hepatocyte growth factor (HGF) is a unique multifunctional growth factor, which has the potential effect to exert neurotrophic action and induce angiogenesis. In this study, we examined the effects of overexpression of human HGF plasmid DNA using ultrasound-mediated gene transfer into the brain in an Aβ-infused cognitive dysfunction mouse model. We demonstrated that HGF gene transfer significantly alleviated Aβ-induced cognitive impairment in mice in behavioral tests. These beneficial effects of HGF might be due to (1) significant recovery of the vessel density in the dentate gyrus of the hippocampus, (2) upregulation of BDNF, (3) a significant decrease in oxidative stress and (4) synaptic enhancement. A pharmacological approach including gene therapy to increase the HGF level in combination with anti-Aβ therapy might be a new therapeutic option for the treatment of AD.Gene Therapy (2008) 15, 561–571; doi:10.1038/sj.gt.3303094; published online 21 February 2008 [ABSTRACT FROM AUTHOR]

The forkhead transcription factor FOXO1, a downstream target of phosphatidylinositol-3-kinase/Akt signalling pathway, regulates cyclic differentiation and apoptosis in normal endometrium, but its role in endometrial carcinogenesis is unknown. Screening of endometrial cancer cell lines demonstrated that FOXO1 is expressed in HEC-1B cells, but not in Ishikawa cells, which in turn highly express the FOXO1 targeting E3-ubiquitin ligase Skp2. FOXO1 transcript levels were also lower in Ishikawa cells and treatment with the proteasomal inhibitor was insufficient to restore expression. Lack of FOXO1 expression in Ishikawa cells was not accounted for by differential promoter methylation or activity, but correlated with increased messenger RNA (mRNA) turnover. Comparative analysis demonstrated that HEC-1B cells proliferate slower, but are more resistant to paclitaxel-mediated cell death than Ishikawa cells, which were partially reversed upon silencing of FOXO1 in HEC-1B cells or its re-expression in Ishikawa cells. We further show that FOXO1 is required for the expression of the growth arrest- and DNA-damage-inducible gene GADD45α. Analysis of biopsy samples demonstrated a marked loss of FOXO1 and GADD45α mRNA and protein expression in endometrioid endometrial cancer compared to normal endometrium. Together, these observations suggest that loss of FOXO1 perturbs endometrial homeostasis, promotes uncontrolled cell proliferation and increases susceptibility to genotoxic insults.Oncogene (2008) 27, 9–19; doi:10.1038/sj.onc.1210626; published online 25 June 2007 [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is highly malignant and prone to multicentric occurrence. Differentiation between a true relapse of HCC and a second primary tumour appearing is of clinical importance. At this point, no convenient method is available to determine the origin of these HCCs. Tissue samples were obtained from 19 patients and analysed for the promoter hypermethylation status of multiple tumour suppressor genes (p16, DAP-Kinase, MGMT, GSTP1, APC, RIZ1, SFRP1, SFRP2, SFRP5, RUNX3, and SOCS1) using methylation-specific PCR (MSP). Methylation status was used to determine tumour clonality. In each of the 19 cases, at least one tumour was recognised as having an aberrantly methylated gene. The frequency of the methylation in tumour tissue was 57.1% in p16, 2.4% in DAP-kinase, 23.8% in GSTP1, 90.5% in APC, 45.2% in RIZ1, 64.3% in SFRP1, 59.5% in SFRP2, 28.6% in SFRP5, 47.6% in RUNX3, and 54.8% in SOCS1, while in MGMT, no aberrant methylation was detected. The methylation status of these genes was assessed using MSP as being either positive or negative, and was used to determine the tumour clonality. The clonality of every tumour could be decided even with lesions that could not be judged by clinical diagnosis or by another molecular method (mt DNA mutation). Determining the methylation status of multiple genes in multicentric HCC was useful as a clonal marker and provided useful information for characterising the tumour. From our findings, multicentric HCCs tend to occur more independently than metastatically from the original tumour. Expanded study should be pursued further for a better understanding of the molecular mechanism of hepatocarcinogenesis. [ABSTRACT FROM AUTHOR]

Using murine models, we have previously demonstrated that recombinant adeno-associated virus (rAAV)-mediated microdystrophin gene transfer is a promising approach to treatment of Duchenne muscular dystrophy (DMD). To examine further therapeutic effects and the safety issue of rAAV-mediated microdystrophin gene transfer using larger animal models, such as dystrophic dog models, we first investigated transduction efficiency of rAAV in wild-type canine muscle cells, and found that rAAV2 encoding β-galactosidase effectively transduces canine primary myotubes in vitro. Subsequent rAAV2 transfer into skeletal muscles of normal dogs, however, resulted in low and transient expression of β-galactosidase together with intense cellular infiltrations in vivo, where cellular and humoral immune responses were remarkably activated. In contrast, rAAV2 expressing no transgene elicited no cellular infiltrations. Co-administration of immunosuppressants, cyclosporine and mycophenolate mofetil could partially improve rAAV2 transduction. Collectively, these results suggest that immune responses against the transgene product caused cellular infiltration and eliminated transduced myofibers in dogs. Furthermore, in vitro interferon-γ release assay showed that canine splenocytes respond to immunogens or mitogens more susceptibly than murine ones. Our results emphasize the importance to scrutinize the immune responses to AAV vectors in larger animal models before applying rAAV-mediated gene therapy to DMD patients.Gene Therapy (2007) 14, 1249–1260; doi:10.1038/sj.gt.3302984; published online 21 June 2007 [ABSTRACT FROM AUTHOR]

Recently, the use of oncolytic viruses against cancer has attracted considerable attention. We studied the potential of the US3 locus-deficient herpes simplex virus (HSV), L1BR1, for oncolytic virus therapy. Its high specificity and potency indicate that L1BR1 is a promising candidate as a new oncolytic virus against pancreatic cancer. Moreover, the virus exhibited the unique characteristic of increasing apoptosis when used in combination with anticancer drugs. We assessed the feasibility of using the US3 locus-deficient HSV named L1BR1 as a new replication-competent oncolytic virus for the treatment of pancreatic cancer. The US3 locus of HSV has been shown to be a key gene in producing a multifunctional protein kinase that inhibits apoptosis induced by viral infections, chemicals and ultraviolet (UV) light. L1BR1 has been reported to be more than 10 000-fold less virulent than the parental virus in mice. In this study, we examined the tumor specificity and oncolytic effect of this attenuated replication-competent virus, L1BR1, in pancreatic cancers derived from SW1990, Capan2 and Bxpc-3cells compared with the parent virus and other well-known oncolytic herpes viruses (R3616 and hrR3). We also studied the efficacy of L1BR1 for the induction of apoptosis as an attribute of this virus in combination with the anticancer drugs 5FU and cisplatin. The combined treatment of the pancreatic cancer cells with L1BR1 and these anticancer drugs enhanced apoptosis significantly. More importantly, L1BR1 showed the lowest replication capacity in normal human hepatocytes, but the highest tumor-reducing effect in vivo among the oncolytic herpes viruses tested. In addition, L1BR1 significantly increased the induction of apoptosis of cancer cells when treated in combination with anticancer drugs although the parental virus inhibited the induction of apoptosis. These results suggest that L1BR1 is promising as a new anticancer oncolytic virus.Cancer Gene Therapy (2007) 14, 533–542. doi:10.1038/sj.cgt.7701049; published online 6 April 2007 [ABSTRACT FROM AUTHOR]

Background. In elderly patients with aneurysmal subarachnoid hemorrhage (SAH), complications including vasosopasm, subdural effusion, and late hydrocephalus, are liable to occur even after aneurysmal surgery. We examined prospectively the efficacy of arachnoid plasty using fibrin glue membrane during surgery of ruptured aneurysms in the elderly patients for preventing complications. The effects on the modified Rankin scale (mRS) and the Glasgow outcome scale (GOS) 3 months after SAH were noted. Methods. Total of 31 patients aged more than 70 years selected from a consecutive series of patients with aneurysmal SAH, were divided into two groups alternately, a group with arachnoid plasty ( n = 16) and a control group without arachnoid plasty ( n = 15). Statistical analyses were performed to assess relationships among various clinical and neuroradiological variables, especially between arachnoid plasty and occurrence of symptomatic vasospasm, subdural effusion, late hydrocephalus, or outcome such as mRS and GOS 3 months after onset. Findings. Statistical analyses revealed that arachnoid plasty were associated with late hydrocephalus and subdural effusion negatively, but with better mRS at 3 months after SAH. A tendency to be associated with less frequent symptomatic vasospasm was also noted. Conclusion. Arachnoid plasty using fibrin glue is suggested to be effective in preventing complications associated with SAH and aneurysmal surgery. A better outcome in the elderly patients can be achieved. [ABSTRACT FROM AUTHOR]

Stage-specific and age-dependent profiles of zinc, copper, manganese, and selenium in testis were examined in Wistar rats by both inductively coupled argon plasma-mass spectrometry (ICP-MS) with a microdissection technique and in situ elemental imaging of micro-PIXE analysis. The young adult animals (10 weeks old) contained higher levels of zinc and manganese in the seminiferous tubules at stages VII-VIII than stages XI through VI and IX-X and the levels were higher than those of the immature and old animals. Copper and selenium levels at stages VII-VIII of the young adult animals were also higher than those of the immature and old animals. In stages VII and VIII, zinc was higher in the central area of the seminiferous epithelium, where spermatozoa were localized, demonstrating a cell-specific property. [ABSTRACT FROM AUTHOR]

BACKGROUND: In pancreatic cancers, K-ras mutations have been found frequently (80%-100%), and they could be a good marker to detect tumor DNA in the plasma. Several studies have indicated that polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis of K-ras mutation was a useful method for the detection of hepatic and lymph node metastasis of pancreatic cancer. However, this method sometimes exhibited false-positive results, and the rate of K-ras mutation might thus be overestimated in these tissues. To diagnose pancreatic cancer correctly at an early stage, we attempted to detect tumor DNA in the plasma of pancreatic cancer patients using a more sensitive and specific method. METHODS: We examined 28 pancreatic cancer patients using a sensitive mutation-specific mismatch ligation assay for K-ras gene mutations in primary tumors and paired plasma samples. RESULTS: K-ras gene mutations were detected in 26 of the 28 (93%) pancreatic cancers. We also found the same mutations in 9 of these 26 (35%) patients in their plasma DNA. This mutation was found even in the plasma of patients with TNM stage II cancer. CONCLUSIONS: Genetic alterations present in the tumors of pancreatic cancer patients can be detected in their plasma, and this approach is potentially applicable for cancer screening and the monitoring of this deadly disease. [ABSTRACT FROM AUTHOR]

Indices of the corpus callosum with normal aging and their sex differences were elucidated using quantitative MRI. We studied 94 Japanese men (mean±SD 57.3±20.8 years, range 6–90 years) and 111 Japanese women (mean±SD 61.2±17.6 years, range 9–86 years) who had no intracranial lesions on MRI and no history of neurological illness. The widths of the rostrum, body and splenium, the anterior to posterior length, and the maximum height in the midsagittal image were selected for measurement. The Evans index, which is the relative ratio of lateral ventricle expansion, and the maximum width of the third ventricle in the axial image were also estimated for comparison. The widths of rostrum, body and splenium of the corpus callosum became thinner with age. Conversely, the anterior to posterior length and the maximum height of the corpus callosum increased with age. The ratio of the width of the body to the length of the corpus callosum and the ratio of the width of the body to the height of the corpus callosum are best correlated with age. No sex differences in regional size of corpus callosum, including these two ratios, were observed in any raw measures, although ventricular indices were larger in men than women. Evaluation of the ratio of the width of the body to its length and the ratio of the width of the body to its height may enable accurate estimation of normal or pathological changes of the corpus callosum. Aging and pathological atrophy of corpus callosum can be evaluated without any adjustment for gender. [ABSTRACT FROM AUTHOR]

Duchenne muscular dystrophy (DMD) is an X-linked, lethal muscular disorder caused by a defect in the DMD gene. AAV vector-mediated micro-dystrophin cDNA transfer is an attractive approach to treatment of DMD. To establish effective gene transfer into skeletal muscle, we examined the transduction efficiency of an AAV vector in skeletal muscles of dystrophin-deficient mdx mice. When an AAV vector encoding the LacZ gene driven by a CMV promoter (AAV-CMVLacZ) was introduced, β-galactosidase expression markedly decreased in mdx muscle 4 weeks after injection due to immune responses against the transgene product. We also injected AAV-CMVLacZ into skeletal muscles of mini-dystrophin-transgenic mdx mice (CVBA3’), which show ameliorated phenotypes without overt signs of muscle degeneration. AAV vector administration, however, evoked substantial immune responses in CVBA3’ muscle. Importantly, AAV vector using muscle-specific MCK promoter also elicited responses in mdx muscle, but at a considerably later period. These results suggested that neo-antigens introduced by AAV vectors could evoke immune reactions in mdx muscle, since increased permeability allowed a leakage of neo-antigens from the dystrophin-deficient sarcolemma of muscle fibers. However, resident antigen-presenting cells, such as myoblasts, myotubes and regenerating immature myofibers, might also play a role in the immune response.Gene Therapy 2002 9, 1576–1588. doi:10.1038/sj.gt.3301829 [ABSTRACT FROM AUTHOR]

Liposomes are micro-compartments made of lipid bilayer membranes withcharacteristics quite similar to those of biological membranes. To formartificial cell-like structures, we generated liposomes that containedsubunit proteins of cytoskeletons: tubulin or actin. Spherical liposomeswere transformed into bipolar or cell-like shapes by mechanical forcesgenerated by the polymerization of encapsulated subunits of microtubules.Disk- or dumbbell-shaped liposomes were developed by the polymerizationof encapsulated actin. Dynamic processes of morphological transformationsof liposomes were visualized by high intensity dark-field lightmicroscopy.Topological changes, such as fusion and division of membrane vesicles,play an essential role in cellular activities. To investigate themechanism of these processes, we visualized in real time the liposomesundergoing topological transformation. A variety of novel topologicaltransformations were found, including the opening-up of liposomes and thedirect expulsion of inner vesicles. [ABSTRACT FROM AUTHOR]

Since the identification in 1987 of the gene for Duchenne muscular dystrophy (DMD), research on the molecular pathogenesis of muscular dystrophy has progressed extensively. In particular, discovery of the DMD gene product, dystrophin, led to the identification of dystrophin-associated proteins and, subsequently, the recognition of other types of muscular dystrophy caused by the defects in each of the sarcoglycan genes. On the other hand, effective therapy for DMD has not yet been established. Some of the viral vectors, such as adeno-associated virus vectors or lentiviral vector, have been proven to enable the long-term expression of the exogenous gene without overt host immune reactions. However, dystrophin cDNAs are too large (14kb) to be accommodated in these viral vectors. To solve this problem, we and other research groups succeeded in truncating full-length dystrophin cDNA to small dystrophin cDNA (4 to 5kb), the products of which protect dystrophin-deficient mdx muscle from contraction-induced membrane damage when introduced by viral vectors or as a transgene into mdx mice. The usefulness of these truncated dystrophin cDNAs should be confirmed using other animal models such as dystrophic dogs. To develop successful treatment of DMD, the authors believe that several different approaches should be used, such as cell transfer therapy, drug design to up-regulate utrophin, or a strategy to repair the mutation in vivo. [ABSTRACT FROM AUTHOR]

To test the hypothesis that 1α-hydroxyvitamin D3 (1α(OH)D3) suppresses bone resorption after ovariectomy (ovx) by inhibiting osteoclastogenic potential in bone marrow cells, the bilateral tibiae of ddY mice, 8 weeks of age, subjected to ovx were obtained. 1α(OH)D3, at doses of 0, 0.2 (low dose), or 0.4 μg/kg body weight (high dose), was administered orally by canula three times a week for 2 or 6 weeks. Histomorphometric analysis of the proximal tibiae revealed that 1α(OH)D3 administration had no significant effect on trabecular bone volume of ovx limbs, which was reduced after ovx. The bone formation rate, increased by ovx, was significantly decreased by the administration of high-dose 1α(OH)D3. The ovx-induced increases in osteoclast number and surface at 2 weeks postsurgery were suppressed by the administration of high-dose 1α(OH)D3. With regard to bone marrow cells, the number of nonadherent cells per tibia obtained from ovx limbs increased, and this increase was suppressed by the administration of low- and high-dose 1α(OH)D3. The formation of mineralized nodules in marrow cultures obtained from ovx limbs was increased after surgery and unaltered by 1α(OH)D3 administration. The number of osteoclast-like multinucleated cells obtained from ovx limbs was reduced by low- and high-dose 1α(OH)D3 administration alike. The number of colony forming units-fibroblast and the number of colony forming units for granulocytes and macrophages was unaltered by ovx or the administration of 1α(OH)D3. The present study clearly demonstrates that high-dose 1α(OH)D3 suppresses osteoclast numbers and surface after ovx. The inhibitory effects of low- and high-dose 1α(OH)D3 on bone marrow cells after ovx were marked in the differentiation from osteoclast precursors to mature osteoclasts. Administration of 1α(OH)D3 suppressed ovx-promoted trabecular bone resorption by inhibiting osteoclastogenic potential in bone marrow cells. [ABSTRACT FROM AUTHOR]

Vertebrates constantly remodel bone to maintain a constant bone mass. Bone remodeling comprises two phases: bone resorption by the osteoclasts followed by bone formation by the osteoblasts. Although the prevailing view about the control of bone remodeling is that it is an autocrine/paracrine phenomenon, the bone resorption arm of bone remodeling is under a tight endocrine control. To date little is known about the regulation of bone formation. We took the observations that gonadal failure favors bone loss and obesity protects from it as an indication that bone mass, body weight, and reproduction could be regulated by the same hormone(s). Leptin is one of these hormones. Leptin inhibits bone formation by the osteoblasts. This function is dominant, and leptin deficiency results in a high bone mass phenotype despite the hypogonadism characterizing these animals. Genetic biochemical and physiological studies demonstrate that leptin inhibits bone formation following its binding to its receptor in the hypothalamus. These results are the first evience that bone remodeling is a hypothalamic process; they imply necessarily that osteoporosis, the most frequent bone remodeling disease, is partly at least a hypothalamic disease. This finding also has therapeutic implications. [ABSTRACT FROM AUTHOR]

Presents a study on the changes in size-fractioned iron concentrations during the growth of the oceanic diatom Chaetoceros specie. Materials and methods; Results and discussion.

Antineutrophil cytoplasmic antibodies (ANCA) have been reported to be associated with systemic vasculitis. However, the roles of ANCA subtypes in patients with IgA nephropathy remain to be fully investigated. We describe three Japanese patients with IgA nephropathy complicated by ANCA-associated vasculitis. Two patients with IgG class ANCA developed rapidly progressive renal failure and demonstrated mesangial proliferation with extensive extracapillary proliferation and segmental glomerular necrosis. One patient with IgM class ANCA showed severe extrarenal symptoms, such as lung fibrosis and neuritis, in addition to glomerular crescent formation. All three patients received immunosuppressive therapies, including corticosteroids and cyclophosphamide. The two patients who received these treatments early showed improvement in urinary protein excretion and renal function, in accordance with a decrease in the serum titer of ANCA. However, one patient in whom serum creatinine was already elevated showed a poor response to the treatment. These results suggest that ANCA subtypes may participate in the pathogenesis of crescent formation in patients with IgA nephropathy, and that early treatment with a combination of methylprednisolone pulse therapy, oral prednisolone, and cyclophosphamide pulse therapy may be beneficial in these patients. [ABSTRACT FROM AUTHOR]

Temperature-dependent electron spin resonance (ESR) measurements of two low-dimensional quantum spin systems, the S = 1/2 spin-ladder system Cu2(1,4-diazacycloheptane)2Cl4 (Cu2(C5H12N2)2Cl4) and the gapless S = 1 bond-alternating one-dimensional antiferromagnet trans-Ni(333-tet)(N3)(ClO4), have been performed. X-band electron spin resonance (ESR) measurements of single-crystal Cu2(C5H12N2)2Cl4 show the increase of linewidth and anisotropic g-shifts below 8 K similar to those known for one-dimensional antiferromagnet. On the other hand, Ni(333-tet)(N3)(ClO4) has broad line-width and ESR has been observed for the first time by our high-field ESR. Its linewidth increases as the temperature is decreased, while the g-shift seems to be isotropic and the g-value decreases as the temperature is decreased. This g-shift can be connected to the quantum fluctuation of the system. [ABSTRACT FROM AUTHOR]

The electric quadrupole interaction of 19O(Iπ=(5/2)+, T1/2=27.0 s) in TiO2 single crystal was studied in detail by means of the β-NQR to determine the electric quadrupole moments Q of short-lived β-emitting nuclei 19O and 13O(Iπ=(3/2)-, T1/2=8.6 ms). Two implantation sites were found for the implanted O nucleus and the quadrupole coupling constants of 19O at these sites were determined. We observed FT-NMR of the enriched stable isotope 17O in TiO2 and obtained the electric field gradient (EFG) at the oxygen substitutional site. With this knowledge, we have determined Q(13O)=11.0 ± 1.3 mb and Q(19O)=3.7 ± 0.4 mb. The present results are compared with the theoretical values calculated by the shell model code, OXBASH and by the Hartree–Fock calculation with the realistic potential. [ABSTRACT FROM AUTHOR]

Background. Macrophage colony-stimulating factor (M-CSF) secreted by malignant cells may regulate cell proliferation via an autocrine and paracrine loop, and M-CSF may be useful as a potential tumor marker. We evaluated the diagnostic significance of M-CSF levels in gynecologic patients. Methods. Serum samples were obtained from 20 healthy women and from 102 gynecologic patients undergoing primary laparotomy, and culture media were obtained from cell lines of gynecologic malignancies (oranian cancer, endometrial cancer, cervical cancer, and choriocarcinoma). M-CSF levels were measured by enzyme-linked immunosorbent assay. Results. M-CSF levels in the sera of patients with ovarian cancer and advanced uterine cancer were significantly higher than those in healthy women and in patients with benign tumors ( P < 0.001). M-CSF levels 7 days after operation were significantly higher than those before operation ( P < 0.001; paired t-test). But there were no differences between M-CSF levels before and after operation in patients with ovarian cancer. M-CSF levels 3 months after the cessation of treatment had decreased to within the normal range in patients with benign tumor and in cancer patients with no residual tumor. Cell lines of gynecologic malignancies produced M-CSF at various concentrations. Conclusion. These findings suggest that M-CSF, produced not only by cancer cells but also by immune cells in ascites and around tumor tissue and by connective tissue may reflect increased serum M-CSF levels in patients with ovarian cancer, advanced uterine cancer, and benign ovarian tumor, associated with ascites. It seems that operation, because of tissue damage, tissue repair, and various physiological responses, may itself, stimulate M-CSF production. [ABSTRACT FROM AUTHOR]

Helice tridens (De Haan) (Grapsidae) and Macrophthalmus japonicus (De Haan) (Ocypodidae) build separate burrows in reed marshes and muddy tidal flats, respectively, in the brackish-water estuaries of northeast Japan (38°11′N, 141°48′E). Habitat segregation between burrows of these two species was analysed by comparing the density of burrows of both species in their natural habitats, and on tidal flats subjected to various types of artificial treatment, in the summer of 1981 and 1982. Baskets, containing many stones, were placed on the ground in the reed marsh, tidal flat and creek and attracted many individuals of H. tridens, but not M. japonicus. When stones were placed on an area of tidal flat, H. tridens frequently formed burrows at the border between the stones and mud, suggesting that burrowing of H. tridens was related to the presence of solid substances lying on the ground, such as stones and shoots of the reed Phragmites australis (Cav.) Trin. In closed systems on the tidal flat and reed marsh, H. tridens and M. japonicus were able to construct burrows in both substrata regardless of high or low frequency. Moreover, it was recognized that H. tridens prevented the burrowing of M. japonicus in these closed systems. These results suggested that habitat segregation between H. tridens and M. japonicus burrows was caused primarily by an exclusive interaction between individuals of these two species. [ABSTRACT FROM AUTHOR]

A highly sensitive radioimmunoassay to measure atrial natriuretic peptide (ANP) concentration in urine has been established, and its clinical usefulness is presented. ANP in urine was stable at 4° C for several days and was easily measured by our radioimmunoassay. The average ANP excretion in 65 healthy persons was 25.0±1.4 ng/day (mean ± SEM) and the fractional excretion of ANP was 0.7±0.05%. In 14 patients with congestive heart failure, the average ANP excretion was 119.2±29.4 ng/day, which decreased to 53.3±11.0 after successful treatment. [ABSTRACT FROM AUTHOR]

In the present study, an attempt was made to clarify whether ANP molecular forms in the plasma of severe congestive heart failure patients differ from those in healthy persons and whether ANP molecular forms in the plasma of the patients were changed by successful treatment of cardiac disease. Twenty patients with congestive heart failure were treated at Kitasato University Hospital. They were classified as class III or IV by New York Heart Association criteria at the time of admission. Plasma ANP concentrations decreased after treatment from 356 ± 58.2 to 72.3 ± 14.8 pg/ml. The gel permeation chromatograms from the plasma of healthy persons showed low, or low and high molecular weight ANP peaks which correspond to the elution positions of authentic α-ANP or ribonuclease A (mol.wt., 13.7 kdalton). In patients with severe congestive heart failure at a severe stage, middle molecular weight ANP consisted with the elution position of authentic β-ANP was particularly noted in addition of low and high molecular weight ANP peaks. This middle molecular weight peak disappears in most of cases by successful treatment. Molecular forms in the plasma obtained from the coronary sinus and the inferior or superior vena cava were essentially the same. These results indicate that the middle molecular weight ANP supposed as β-ANP may particularly be secreted in severe congestive heart failure patients. [ABSTRACT FROM AUTHOR]

Effects of molybdenum and tungsten on carbon activity in nickel have been experimentally determined at 1000, 1100, and 1200 °C. Seventeen nickel-molybdenum and thirteen nickel-tungsten binary alloys were carburized in a flow of purified methane and hydrogen mixed gas. A sealed capsule technique was also employed for carburization of a few series of nickel-molybdenum alloys. The carbon concentration was determined either by hot extraction techniques (LECO and Coulomatic) or by weight gains of these specimens. The carbon concentration at a constant carbon activity decreases with increasing either molybdenum or tungsten concentration in nickel. The effect of tungsten on the carbon solubility in nickel is slightly larger than that of molybdenum. The experimental data were analyzed in terms of the regular solution model with two sublattices due to Hillert and Staffansson. Temperature dependence of the interaction coefficients between carbon and molybdenum or tungsten was expressed as DG/RT = −4.45 + 11650/ T and DG /R = 1.21 + 9010/. [ABSTRACT FROM AUTHOR]

Paneth cells located at the bottom of intestinal crypts may play a role in controlling the bacterial milieu of the intestine. Using morphometry to clarify the secretory mechanism of the Paneth cells, we studied the ultrastructural changes in mouse Paneth cells produced following intra-arterial perfusion with Hanks' balanced salt solution containing a cholinergic muscarinic secretagogue (bethanechol), a neuroblocking agent (tetrodotoxin), or a G-protein activator (NAF/AlCl). Bethanechol (2×10 mol/ l) induced Paneth-cell secretion. Many Paneth cells massively exocytosed their secretory material into the crypt lumen; the enhanced secretion caused degranulation and vacuole formation. However, tetrodotoxin (2×10 mol/ l) did not prevent the bethanechol-enhanced secretion by the Paneth cells. NaF (1×10 mol/ l) and AlCl (1×10 mol/ l) induced massive exocytosis of the Paneth cells; the exocytotic figures were similar to those observed in mice stimulated by bethanechol. G-protein activation was followed by a sequence of intracellular events, resulting in exocytosis. [ABSTRACT FROM AUTHOR]