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The cooling effect of oil cooling method on electric vehicle motors with hairpin winding is analyzed. A 160 kW motor being developed by a commercial vehicle company is applied. Firstly, the flow field of oil was analyzed. In the crown part, 96% of the oil flowed; in the welded part, 90% of the oil flowed. Secondly, an analysis of the heat transfer in the coil and stator was performed. The welded part of the coil exhibited an average temperature 4% higher than that of the crown part. Finally, the parametric study was conducted to understand the effect of the temperature and flow rate of the oil on the temperature distributions of the motor. As the flow rate increases, the maximum temperature decreases. However, there is not a big difference at more than 0.8 LPM/LPM. The results provide important information for understanding the effect of a motor using oil cooling.

Cutaneous melanoma is known to be one of the most dangerous skin cancers because of its metastatic functions. Today, it is essential to investigate specific biomarkers for the target treatment in many diseases including cancers. DJ-1 protein, also known as Parkinson disease 7, has various functions associated with cancer progression including cell survival and migration. Phosphatase and tensin homolog (PTEN) is a tumor suppressor that regulates the PI3K/AKT signaling pathway and its mutations have been reported to frequently occur in many cancers such as thyroid, breast and skin. Recently, DJ-1 has been identified as a negative regulator of PTEN in many human cancer cells. However, the impacts and relationship of DJ-1 and PTEN have not been studied yet in melanoma. To confirm the expression of DJ-1 and PTEN in melanoma compared to normal skin tissues and find out functions of DJ-1 in melanoma cells, Western blot analysis and immunohistochemical staining were used. Transfection of G361 cells with DJ-1-specific small interfering RNA was performed to figure out the roles of DJ-1 and the relationship between DJ-1 and PTEN in melanoma cells. In our study, the DJ-1 protein was significantly increased with loss of PTEN protein in melanoma compared to that in normal skin. Inhibition of DJ-1 in G361 cells induced apoptosis, and suppressed cell survival and migration. Furthermore, suppression of DJ-1 in G361 cells increased the expression of cleaved caspase-3, cleaved PARP, Bax, p53, and Daxx as well as PTEN, while it decreased expression of survivin, caspase-3, and PARP. Also, downregulated DJ-1 inhibited phosphorylation of AKT in G361 cells. Collectively, DJ-1 overexpression could affect the proliferative and invasive capabilities of melanoma cells via regulating the PTEN/AKT pathway and apoptosis-related proteins. This study suggests that DJ-1 may be a potential target for the treatment of melanoma.

Arctigenin derived from the seeds of Arctium lappa Linnaeus is known as an anticancer drug candidate by targeting various pathways involved in anticancer therapy. Using 2D monolayer and 3D spheroid culture systems in nasal septum carcinoma RPMI-2650 cells, the effects of arctigenin and dexamethasone on cell viability, ROS levels, ATP level, mitochondrial function, apoptosis and necroptosis were examined. The combination treatment of both compounds induced strong cytotoxicity, accompanied by increases of sub-G0/G1 peak, annexin V-PE-positive cells, and ROS levels, loss of mitochondrial membrane potential, and decrease of cellular ATP content. These changes were observed as simultaneous induction of DNA damage, apoptosis, and necroptosis. A series of changes by arctigenin and dexamethasone were efficiently restored by decreasing ROS levels or supplementing ATP. Treatment of 3D spheroids with arctigenin and dexamethasone decreased cell viability in the spheroids, but it was slightly resistant than cells under 2D conditions. In addition, this phenomenon was accompanied by an increase in mediators for both apoptosis and necroptosis. Results of this study suggest that the apoptosis and necroptosis-inducing effects of arctigenin are associated with ATP depletion due to oxidative mitochondrial dysfunction.

Separability is an attractive feature of covariance matrices or matrix variate data, which can improve and simplify many multivariate procedures. Due to its importance, testing separability has attracted much attention in the past. The procedures in the literature are of two types, likelihood ratio test (LRT) and Rao’s score test (RST). Both are based on the normality assumption or the large-sample asymptotic properties of the test statistics. In this paper, we develop a new approach that is very different from existing ones. We propose to reformulate the null hypothesis (the separability of a covariance matrix of interest) into many sub-hypotheses (the separability of the sub-matrices of the covariance matrix), which are testable using a permutation based procedure. We then combine the testing results of sub-hypotheses using the Bonferroni and two-stage additive procedures. Our permutation based procedures are inherently distribution free; thus it is robust to non-normality of the data. In addition, unlike the LRT, they are applicable to situations when the sample size is smaller than the number of unknown parameters in the covariance matrix. Our numerical study and data examples show the advantages of our procedures over the existing LRT and RST.

The Korea Research Institute of Standards and Science (KRISS) participated in the BIPM.RI(II)-K4.Tc-99 m comparison of activity measurements of 99mTc. We develop the national primary standard for 99mTc solution used during the comparison. We use two primary liquid scintillation counting techniques for the standardization: digital coincidence counting with the 4πβ(LS)-γ(NaI(T1))system and triple-to-double coincidence ratio counting. Both methods give consistent results for the specific activity of a 99mTc solution. Adopting the result from the DCC method as the reference value of the 99mTc measurement, we evaluate the standard uncertainty of 0.856% for the KRISS primary standard of the 99mTc activity. The K4 comparison result shows that the newly-established KRISS primary standard lies within the standard uncertainty with the key comparison reference value defined from other 99mTc measurements.

In this study, the high-pressure steaming and enzymatic pre-treatment (SET) was used to improve β-glucan extraction from Ganoderma lucidum (G. lucidum), an oriental medicinal mushroom. Response surface methodology and central composite design were used to determine the optimum pre-treatment conditions: high-pressure steaming, enzymatic hydrolysis, and Viscozyme L concentrations. The optimal conditions were 15.51 min for high-pressure steaming, 0.84 g/100 mL of Viscozyme L, and 4.16 h for hydrolysis. The predicted β-glucan content in G. lucidum extract at optimal conditions, approximately twofold (8.05 g/100 g) of the control treatment value, was consistent with the empirical value. The total sugar and protein contents through SET were higher than those values of the control treatment. The cell migration assay showed that SET-processed G. lucidum extracts significantly suppressed B16F10 murine melanoma cell growth. SET process using Viscozyme L could be utilized for β-glucan extraction from G. lucidum to develop the functional food.

Reactive oxygen species (ROS) contribute to pathogenesis of many inflammatory skin diseases, including psoriasis. The aim of this study is to compare antioxidant protein expression in psoriasis vulgaris (PV) skin tissues with that in normal skin tissues in vivo and to evaluate the effects of dimethyl fumarate (DMF), used for the treatment of psoriasis, on ROS generation and apoptosis in a human keratinocyte cell line HaCaT. Compared with normal skin tissues, PV skin tissues showed increased protein oxidation as well as down-regulation of Nrf2 and its regulatory proteins such as HO-1 and AKR1C3. Using HaCaT cells to model DMF-induced pro-oxidant effects in the skin cells, we found that DMF treatment induced increased ROS levels and apoptotic cell death, as signified by increased proportion of cells with Annexin V-PE(+) staining and a sub-G0/G1 peak in the cell cycle. Preceding these changes, DMF treatment resulted in up-regulation of Nrf2, HO-1, and AKR1C3 proteins in these cells. Collectively, increased oxidative stress and impaired cellular anti-oxidant enzyme systems may participate in the pathogenesis of PV. DMF may exert an additive therapeutic efficacy in PV by attenuating the redox burden and subsequent oxidative damage to normal keratinocytes through activation of Nrf2 pathway relative to PV.

Functional defects in apoptosis signaling have been considered the important mechanism of chemoresistance in malignant mesothelioma (MM). This study was designed to explore the growth-inhibiting and apoptosis-promoting effects of sulforaphane and cisplatin on the human MM cell lines, MSTO-211H and H-2452, and examine the underlying mechanisms. Compared to normal mesothelial Met-5A cells, sulforaphane and cisplatin exhibited synergistic anti-cancer efficacy towards MM cells. By inducing ROS accumulation and mitochondrial dysfunction, sulforaphane potentiated cisplatin-induced cytotoxic effects, as evidenced by the increased nuclear fragmentation and chromatin condensation with the enhanced cleavage of procaspase-3 and PARP, the increased percentage of Annexin V-PE (+) cells, and the appearance of a sub-G0/G1 peak in the flow cytometric analysis. The apoptosis-inducing effects of two compounds were associated with the increased Bax/Bcl-2 ratio due to a decrease in the anti-apoptotic Bcl-2 level. A G2/M phase transition delay in the cell cycle following the combined treatment of sulforaphane and cisplatin was linked to the down-regulation of cyclin B1 and phospho-Cdc2 (Thr161) levels. In addition, the inhibition of ROS with N-acetylcysteine attenuated the apoptosis induced by the combination treatment, and recovered the depletion of mitochondrial membrane potential, revealing the vital role of ROS in the synergism. By targeting redox homeostasis, such a pro-oxidant-based combinational approach will help to enhance the therapeutic efficacy and preferential cytotoxicity on MM cells that conventional therapy would be ineffective.

Intra-articular injection of local anesthetics (LAs) is a common procedure for therapeutic purposes. However, LAs have been found toxic to articular cartilage, and hyaluronan may attenuate this toxicity. In this study we investigated whether hyaluronan attenuated lidocaine-induced chondrotoxicity, and if so, to elucidate the underlying mechanisms. Human chondrocyte cell line SW1353 and newly isolated murine chondrocytes were incubated in culture medium containing hyaluronan and/or lidocaine for 72 h. Cell viability was evaluated using MTT assay. Cell apoptosis was detected with DAPI staining, caspase 3/7 activity assay and flow cytometry. Cell cycle distributions, ROS levels and mitochondrial membrane potential (ΔΨm) were determined using flow cytometry. The expression of p53 and p53-regulated gene products was measured with Western blotting. Lidocaine (0.005%−0.03%) dose-dependently decreased the viability of SW1353 cells. This local anesthetic (0.015%, 0.025%) induced apoptosis, G2/M phase arrest and loss of ΔΨm, and markedly increased ROS production in SW1353 cells. Hyaluronan (50−800 μg/mL) alone did not affect the cell viability, but co-treatment with hyaluronan (200 μg/mL) significantly attenuated lidocaine-induced apoptosis and other abnormalities in SW1353 cells. Furthermore, co-treatment with lidocaine and hyaluronan significantly decreased the levels of p53 and its transcription targets Bax and p21 in SW1353 cells, although treatment with lidocaine alone did not significantly change these proteins. Similar results were obtained in ex vivo cultured murine chondrocytes. Hyaluronan suppresses lidocaine-induced apoptosis of human chondrocytes in vitro through inhibiting the p53-dependent mitochondrial apoptotic pathway.

Quercetin, a naturally occurring flavonoid, has been heralded as a promising chemopreventive agent. This study was undertaken to probe the molecular mechanisms underlying the anti-cancer activity of quercetin in malignant mesothelioma (MM) cells. Quercetin at low doses elicited apoptotic cell death on MM MSTO-211H cells, as signified by pyknotic and fragmented nuclei, increased annexin V binding, and increased proportion of cells with hypodiploid DNA. Preceding these changes, quercetin induced up-regulation of p53 at both mRNA and protein levels without altering its ubiquitination, and increased caspase-3/7 activity with the resultant cleavages of procaspase-3 and PARP. Analyses of nuclear p53 level, p53 reporter gene, and RT-PCR toward p53-regulated genes demonstrated that induced p53 is transcriptionally active. The proportion of cells at sub-G0/G1 peak and G2/M phase increased in a quercetin concentration-dependent fashion, which were blocked by the pan-caspase inhibitor Z-VAD. Additionally, quercetin and gemcitabine produced a significant synergistic effect on inhibiting MS-TO-211H cell growth. Given that quercetin induced preferential p53-upregulating, growth-inhibiting, and apoptosis-activating effects on MM cells, the use of quercetin may be a potential therapeutic strategy for enhancing anti-cancer efficacy of existing chemotherapy in MM.

The aim of the present study was to examine the antioxidative activity of (+)-lariciresinol (LRSL), an optically active lignan isolated from Rubia philippinensis in several in vitro assays. LRSL was also subjected to evaluate its inhibitory effect against the generation of reactive oxygen species (ROS) in murine macrophage (RAW 264.7) cells. The results showed that LRSL possessed very strong radical scavenging activity and reducing power, as well as inhibited ROS generation in a dose-dependent manner without showing any cytotoxicity. The transcriptional and translational levels of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were markedly higher in the sample treated group. LRSL treatment also increased the transcriptional and translational activities of NF-E2-related factor-2 (Nrf-2) with a corresponding increase in the transcriptional and translational activities of the heme oxygenase-1 (HO-1). LRSL activated p38 and treatments with SB239063 (a p38 inhibitor) suppressed the LRSL-induced activation of Nrf2, resulting in a decrease in HO-1 expression. Collectively, the data demonstrated that LRSL has potent antioxidative activity, decreasing ROS generation in RAW 264.7 cells and increasing the transcriptional and translational levels of antioxidant enzymes by activating Nrf2-mediated HO-1 induction via p38 signaling.

Traditionally in bioassay monitoring, 3H determinations in urine have been performed using either direct counting (with or without sample decolourisation) or distillation whilst 14C has been determined on separate sub-samples following chemical isolation of the carbon from samples. Although these techniques are relatively straightforward they do not permit simultaneous determination of 3H/14C. For 3H, direct counting can be significantly affected by variations in sample composition/colour resulting in varying degrees of sample quench, does not distinguish between aqueous/organically-bound 3H and is limited to small sample sizes. This study describes the use of purpose built multi-tube combustion furnace for the simultaneous extraction and determination of 3H and 14C. The technique is insensitive to sample composition and can be adapted to measure Tritiated water (HTO) and organically bound tritium separately. The development of a multi-tube system with integrated cool-down facility permits rapid throughput of high sample numbers and has been proven effective in decommissioning waste characterisation. In addition, the furnace-based technique is capable of processing larger sample sizes, increasing analytical sensitivity and accuracy of dose assessment.

Nasal epithelium is the first cellular target for inhaled cadmium; however, the specific molecular events that mediate or accompany toxicity are not well studied. Using RPMI-2650 cells as an in vitro nasal epithelial model for the investigation of molecular events of cadmium toxicity, we found that treatment with cadmium acetate induced cell death with the typical morphological changes of apoptosis, such as chromatin condensation and nuclear fragmentation, as well as increased caspase-3/7 activity. This finding was accompanied with concentration-dependent increases of RUNX3, phospho-p53 (p-p53), and phospho-Akt (p-Akt) proteins with concomitant decrease in the Bcl-2 level. However, no significant changes were observed in the levels of Fas ligand and Fas-associated protein with death domain. Cadmium treatment increased the DNA binding activity of p53 toward a p53 consensus binding site. Blockage of RUNX3 expression with siRNA transfection did not affect a cadmium-induced increase in the p-p53 level but suppressed p53 binding activity to DNA. Nuclear co-localization of RUNX3 and p-p53 was enhanced by treatment with cadmium. The nuclear RUNX3 level was suppressed by the pretreatment with LY294002, a PI3-kinase inhibitor, while noticeable changes in the levels of p53 and p-p53 were not observed under the same condition. Cadmium-induced increase in the p53 binding to DNA was partly suppressed by LY294002, while it was enhanced by PD98059, a MEK inhibitor. Collectively, our findings suggest that cadmium induces up-regulation of RUNX3 protein through activation of PI3-kinase/Akt and leads to RPMI-2650 cell death, at least in part, through p53-dependent apoptosis pathway, in which RUNX3 may facilitate p53 binding to DNA.

A straightforward and rapid method has been developed for the determination of 241Pu activities. Pu is chemically separated from the sample, purified and electrodeposited to produce a source for alpha spectrometric determination of 238Pu and 239,240Pu. Pu is stripped from the disc with concentrated nitric acid and extracted into tri-octylphosphine oxide (TOPO)/toluene. The organic extract is then mixed directly with commercial liquid scintillation cocktail without any further purification procedures and the sample counted on a Wallac 1220 Quantulus liquid scintillation counter (LSC). 241Pu activity is estimated via the 242Pu yield monitor acquired by alpha spectrometry measurement. Experimental results for the performance testing of a low-level liquid scintillation spectrometer and the data for the evaluation of the method using standard reference materials are presented.

This study was investigated the association between Angiotensin Converting Enzyme (ACE) polymorphism on blood lead levels and lead-related blood pressure of exposed male workers from Korea. A cross sectional study involving 786 male lead-exposed workers from Korea was conducted, in which a polymerase chain reaction (PCR)-based strategy was used to differentiate the genotypes of ACE I/D (Insertion/ Deletion) polymorphism. The association of clinical characteristics with genotypes as modifiers was estimated after data adjustment for age, smoking status, drinking status, body mass index and job duration. Genotype and allele frequencies of ACE I/D polymorphism were associated with lead-related diastolic, systolic blood pressure and hypertension status. Blood lead levels were not associated with genotype or allele frequencies of ACE I/D polymorphism. From the present results, it was concluded that I/I (Insertion/Insertion)+ I/D genotypes, and I allele of the ACE gene is a risk factor for lead-related hypertensive status; however, they are not directly associated with blood lead levels.

Radionuclides such as 131I, 134Cs, 137Cs, and 239,240Pu in Korean rainwater have been analyzed by Korea Research Institute of Standards and Science (KRISS) since the Fukushima nuclear power plant accident in March 2011 to investigate the activity level, distribution pattern, and temporal variation and to assess the radiation dose the public is exposed to. The concentration of 131I in the Korean rainwater samples varied between 0.033 (minimum detectable activity; MDA) and 1.30 Bq kg−1 and the concentrations tended to decrease exponentially with time. The concentrations of 134Cs and 137Cs in rainwater ranged from 0.01 to 334 ± 74 and 0.29 ± 0.01 to 276 ± 1 mBq kg−1, respectively. The mean activity ratio of 137Cs/134Cs in the rainwater samples collected from April 18 to May 12 was estimated to be 0.44 ± 0.21, and this value is lower than that (ca. 1) observed in Fukushima, Japan, when there was an escape from the nuclear reactors. When an attempt was made to analyze Pu isotopes in rainwater samples, no Pu isotopes were detected above the MDA in any of the rainwater samples. Although the locations investigated were different from Asia to Europe, the concentrations of 131I, 134Cs and 137Cs in the rainwater are comparable, which suggests a global contamination of 131I, 134Cs, and 137Cs occurred because of the Fukushima nuclear power plant accident.

137Cs was measured in soil samples collected in Korea from 2006 to 2008 using a low-level background gamma-ray spectrometer that was designed and developed by KRISS (Korea Research Institute of Standards and Science). The objectives of this study are to evaluate the newly developed low-level background gamma-ray spectrometer and, consequently, to provide information on the horizontal and vertical distribution of 137Cs in Korean soil. 137Cs concentrations in surface soil varied from 12.8 ± 0.9 to 108 ± 4 Bq kg−1, and the vertical profiles of 137Cs from the Nari basin in Ulleung Island in the East Sea/Sea of Japan and Seongsan Ilchulbong Peak, Jeju Island, showed a higher concentration in the surface layer that gradually decreased with depth. On the other hand, the 137Cs concentration in soil samples collected from Bukhan Mountain National Park in Seoul showed a subsurface maximum and decreased with depth. The 137Cs inventories in the soil column were calculated to be 1,830–4,360 Bq m−2 with a mean of 2,770 Bq m−2, which was the same order of magnitude as the global fallout inventories in the mid-latitude region of the Northern Hemisphere.

Quercetin, a member of the flavonoids, has been heralded as chemopreventive agent, generally because of its free radical-scavenging and antiproliferative activities. In this process, NF-E2-related factor-2 (Nrf2) plays a role in protecting cells from oxidative damage; however, the exact mechanisms in which bronchial epithelial cells respond to quercetin by activating Nrf2 are not fully understood. Here, we report the role of extracellular signal-regulated kinase (ERK) activation in response to quercetin. Quercetin treatment decreased the viability of human bronchial epithelial BEAS-2B cells in a dose- and time-dependent manner. ERK1/2 was activated within 10 min of quercetin addition and then remained increased above control level throughout 48 h, whereas AKT phosphorylation was suppressed from the first 1 h of quercetin treatment. Inhibition of Erk1/2 phosphorylation attenuated quercetin-induced loss of cell viability. Nrf2 levels in both nuclear and whole cell lysates increased after quercetin treatment and was accompanied by the increase of heme oxygenase-1 (HO-1) protein level, indicating that increased HO-1 expression is Nrf2-mediated. Activation of the Nrf2/HO-1 system after quercetin treatment was suppressed by the pretreatment with MEK inhibitor, PD98059. Also, silencing Nrf2 with siRNA decreased cell viability and augmented cytotoxic effect of anticancer drug LBH589. Overall, our results indicate that quercetin induces the up-regulation of Nrf2-mediated HO-1 expression, at least in part, via MEK/ERK1/2 signaling, as an adaptive mechanism which may provide advantages for cell proliferation and enhance survival despite endogenous and/or exogenous oxidative insults.

Unripe apples lie scattered about the orchards because of manual thinning out or falling, however they contain a high amount of polyphenol. In order to enhance the extraction of polyphenol from the unripe apples, carbohydrate-hydrolyzing enzyme (Viscozyme L) aided polyphenol extraction techniques have been studied with response surface methodology. The optimum conditions were as follows: the ratio of Viscozyme L to substrate was 0.0195 (1.95 fungal beta-glucanase units), the reaction temperature was 47.12°C, and the reaction time was 12.52 h. The experimental values of total phenolic content and caffeic acid content of the unripe apples were well matched with the response surface methodology predicted values. The levels of total phenolic content and caffeic acid content obtained from Viscozyme L treatment were about 2 and 13 folds greater, respectively than those of the control treatment.

Analysis of plutonium isotopes by Semiconductor Alpha Spectrometry (SAS), ICP-sector field mass spectrometry (ICP-MS) and Accelerator Mass Spectrometry (AMS) was carried out in seawater samples collected from the Northeast Atlantic Ocean (nuclear waste dumping sites) and Northwest Pacific Ocean. No particularly elevated levels of the atom ratios of 240Pu/239Pu compared to global fallout ratio (0.18) were found in the Northeast Atlantic Ocean seawater samples. The higher levels of atom ratios of 240Pu/239Pu were found in the Northwest Pacific Ocean. This is mainly due to contribution from the local fallout from nuclear weapon tests carried out at the Pacific Proving Grounds at the Marshall Islands.

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Gastaud, Janine Oregioni, Beniamino Coppola, Laurent Jeandel, Catherine 5 SPRINGER DORDRECHT J OCEANOGR; Anthropogenic Sr-90, Pu-239,Pu-240 and Am-241 were used as tracers of water mass circulation in the Crozet Basin of the South Indian Ocean, represented by three main water fronts-Agulhas (AF), Subtropical (STF) and Subantarctic (SAF). Higher Sr-90 concentrations observed north of 43A degrees S were due to the influence of AF and STF, which are associated with the south branch of the Subtropical gyre, which acts as a reservoir of radionuclides transported from the North to the South Indian Ocean. On the other hand, the region south of 43A degrees S has been influenced by SAF, bringing to the Crozet Basin Antarctic waters with lower radionuclide concentrations. The Pu-238/Pu-239,Pu-240 activity ratios observed in water and zooplankton samples indicated that, even 35 years after the injection of Pu-238 to the Indian Ocean from the burn-up of the SNAP-9A satellite, the increased levels of Pu-238 in surface water and zooplankton are still well visible. The radionuclide concentrations in seawater and their availability to zooplankton are responsible for the observed Po-210, Pu-239,Pu-240 and Am-241 levels in zooplankton.

Withaferin A, a major chemical constituent of Withania somnifera, has been reported for its tumor cell growth inhibitory activity, antitumor effects, and impairing metastasis and angiogenesis. The mechanism by which withaferin A initiates apoptosis remains poorly understood. In the present report, we investigated the effect of withaferin A on the apoptotic pathway in U937 human promonocytic cells. We show that withaferin A induces apoptosis in association with the activation of caspase-3. JNK and Akt signal pathways play crucial roles in withaferin A-induced apoptosis in U937 cells. Furthermore, we have shown that overexpression of Bcl-2 and active Akt (myr-Akt) in U937 cells inhibited the induction of apoptosis, activation of caspase-3, and PLC-gamma1 cleavage by withaferin A. Taken together, our results indicated that the JNK and Akt pathways and inhibition of NF-kappaB activity were key regulators of apoptosis in response to withaferin A in human leukemia U937 cells.

We have investigated the effects of BRCA1 over-expression and knockdown on 5F-203-induced gene expression and cytotoxicity in human breast cancer cells. 5F-203 is a chemotherapeutic prodrug that both induces a p450 enzyme, CYP1A1, and is metabolically activated by CYP1A1.We used several molecular biological techniques to confirm our findings. BRCA1 regulates sensitivity to 5F-203 by regulating the expression of CYP1A1 mRNA and its EROD activity. XRE-Luc reporter assays, semi-quantitative RT-PCR, Western blot analysis, EROD activity measurements, gene knockdown and MTT cell survival assays were used for this study.Our results show that the ability of 5F-203 treatments to increase CYP1A1 mRNA level and CYP1A1 enzymatic activity (EROD activity) are affected by BRCA1 protein levels. In addition, the ability of 5F-203 treatments to induce proteins, P53 and P53 target genes such as P21, is significantly decreased in BRCA1 knockdown cells, suggesting that BRCA1-related effects could at least partially explain why BRCA1 knockdown increases resistance to 5F-203-mediated cytotoxicity. We also observed altered expression of the two major transcription factors (AhR and ARNT) that affect CYP1A1 expression when BRCA1 protein levels are altered.BRCA1 is an important protein, which affects 5F-203-mediated cytotoxicity. Our findings are potentially clinically significant; they suggest that those patients most likely to respond to this new prodrug will have tumors containing normal amounts of BRCA1.

The requirements of multi-resolution models of feature-based solids, which represent an object at many levels of feature detail, are increasing for engineering purposes, such as analysis, network-based collaborative design, virtual prototyping and manufacturing To provide multi-resolution models for various applications, it is essential to generate adequate solid models at varying levels of detail (LOD) after feature reanangement, based on the LOD criteria However, the non-commutative property of the union and subtraction Boolean operations is a severe obstacle to arbitrary feature rearrangement To solve this problem we propose history-based Boolean operations that satisfy the commutative law between union and subtraction operations by considering the history of the Boolean operations Because these operations guarantee the same resulting shape as the original and reasonable shapes at the intermediate LODs for an arbitrary rearrangement of its features, various LOD criteria can be applied for milti-resolution modeling in different applications

241Pu was measured in reference materials and marine samples using a novel method based on rare earth fluoride co-precipitation followed by liquid scintillation spectrometry (LSS). Disc sources used for α-spectrometry of 238Pu and 239+240Pu were leached with concentrated nitric acid before neodymium fluoride co-precipitation to scavenge plutonium in the lower oxidation states was carried out. After the determination of the chemical recovery by α-spectrometry, the precipitate was recovered in a H3BO3/HNO3 based dissolver and mixed with Packard Insta-GelO Plus cocktail before LSS. Reasonable agreement has been obtained between the results obtained using this methods and the data obtained by direct disc analysis by LSS and ICP-MS. The 241Pu/239+240Pu ratios were coherent with the expected origin of the Pu contaminant in the marine environment showing the validity of the method.

Formation of Fe(II)-containing mineral through microbial processes may play an important role in iron and carbon geochemistry in subsurface environments. Fe(III)-reducing bacteria form Fe(II)-containing minerals such as siderite, magnetive, vivianite, and green rust using iron oxides. A psychrotolerant Fe(III)-reducing bacterium,Shewanella alga (PV-4), was used to examine the reduction and biomineralization of a poorly crystalline iron oxide, akaganeite (β-FeOOH), in the absence of a soluble electron shuttle, anthraquinone disulphonate (AQDS), under different atmospheric compositions as well as in HCO 3 − buffered medium (30 to 210 mM). Iron biomineralization was also examined under different growth conditions such as incubation time, electron donors, and electron acceptors. The Fe(III)-reducing bacterium, PV-4, reduced akaganeite, Fe(III)-citrate, and Co(III)-EDTA using lactate or H2 as an electron donor. The iron biomineralization of Fe(III) oxide, akaganeite—as it undergos reduction by an iron reducing bacterium—is a complex process influenced by biogeochemical factors including microorganisms, bicarbonate buffer concentration, atmospheric composition, electron donors/acceptors, incubation time, and Eh/pH. From this research we found that microorganisms do participate in the formation of diverse iron minerals and that microbial iron biomineralization may affect Fe and C biogeochemistry in subsurface environments.

Radionuclides in aerosols and precipitation have been analyzed by IAEA-MEL in Monaco since 1987 and 1999, respectively, to identify their behavior in the atmosphere and delivery into the northwestern Mediterranean Sea. While the concentration of 7Be in aerosols has been affected by the stratospheric-tropospheric mixing and precipitation, the concentration of 137Cs in aerosols has been influenced by a combination of local meteorological conditions and re-suspension of 137Cs from soil. Higher concentrations of 7Be, 137Cs, 239,240Pu and 241Am in precipitation occurred during spring and summer. The highest deposition rates of these radionuclides were observed in spring and autumn during periods of high precipitation. A good correlation was found between the amount of precipitation and depositional flux for 7Be, but not for 137Cs, 239,240Pu and 241Am. This indicates that the wet process as well as the dry process seem to be important factors in determining the annual deposition for transuranic radionuclides and 137Cs.

In this paper we determine the general solution of the functional equation¶¶f(ax + by + cxy) = p(x) + q(y) + g(x)h(y)¶which is a generalization of the Pompeiu functional equation.

Aberrant transcripts of FHIT (fragile histidine triad) have been reported in several types of primary tumors and cell lines, including gastric carcinoma. The role of these aberrant transcripts in tumorigenesis is not clear yet. Forty-eight aberrant-sized FHIT transcripts with various lengths and number in 35 cases of gastric adenocarcinomas were further characterized. Aberrant transcripts, with deletions and/or insertions, were frequently observed in 20 cases of tumors. Sequence analysis demonstrated that different types of aberrant transcripts used normal splice sites but skipped exons, contained the inserts with the part of intron 5 sequences, or used the FHIT cDNA sequence 179-180 as a cryptic splice acceptor site. Most of aberrant transcripts lacked exon 5 and were presumably non-functional as the translation initiation codon is located in exon 5. Additionally, other transcripts, indicative of additional splice processing, either deletions or insertions, were expressed in several tumors. Taken together, our data indicate that the FHIT gene expression is frequently altered in gastric adenocarcinomas by aberrant splicing, and suggest that different types of aberrant transcripts may result during the multi-step splice processing.

Liquid scintillation spectrometry has become the most widespread method for quantitative analysis of low level β-emitters in environmental samples. This technique has been applied in the measurements of 241Pu, 3H and 90Sr in seawater and sediment samples. 241Pu can be measured by direct analysis of an electrodeposited source using α-β discrimination or by extraction of electrodeposited plutonium into a liquid form compatible with scintillation cocktail. Sediment from Mururoa and Fangataufa atolls showed activities ranging from 18 to 44 Bq/kg. A sediment profile sampled around Bikini Atoll in 1997 showed 241Pu activities ranging from 0.3 to 30 Bq/kg. 3H activities in pore water sediment from Mururoa and Fangataufa atolls were of the order of 103 Bq/m3 which demonstrated its underground origin. 90Sr was measured in the presence of 90Y. The counting efficiency was 92.41.5% and the background 0.027±0.001 s-1. The average chemical recovery for 90Sr was 673%.

The most important recent developments in radiometric techniques have been the operation of high efficiency HPGe detectors with anticosmic or antiCompton shielding often placed underground, ship-board measurements of 234Th using gamma-spectrometry or beta-counting and underwater gamma-spectrometry. In mass spectrometry techniques, the availability of high resolution ICP-MS and applications of AMS for the analysis of long-lived radionuclides have opened doors for investigations which previously required too large samples, or were not possible because of lack of sensitivity. These developments are illustrated by several examples of marine radioactivity studies, which include contaminated sites (e.g., Fangataufa Atoll) and open ocean studies (e.g., the Atlantic Ocean) as well as analyses of IAEA reference materials. A comparison of Pu results obtained by alpha-spectrometry, ICP-MS and AMS has shown that a reasonably good agreement has been obtained between all three methods.

IAEA reference materials (radionuclides in the marine environment) collected in areas affected by nuclear reprocessing plants and nuclear weapons tests have been analysed by semiconductor alpha-spectrometry (SAS), liquid scintillation spectrometry (LSS) and mass spectrometric techniques (high resolution ICP-MS and AMS) with the aim of developing analytical procedures and to study the geochemical behavior of plutonium in the marine environment. The Pu results obtained by SAS, ICP-MS and AMS were in reasonably good agreement (R 2 = 0.99). The mean atom ratios of 240Pu/239Pu in IAEA reference materials, IAEA-134, 135 and 381 were (0.212±0.010), (0.211±0.004) and (0.242±0.004), respectively. IAEA-384 (Fangataufa Lagoon Sediment) gave a 240Pu/239Pu mean atom ratio of 0.051±0.001. The results of 241Pu obtained by ICP-MS and LSS also show reasonable agreement (R 2 = 0.91). Pu isotopic signatures were useful in tracing Pu origin and in interpreting biogeochemical processes involving Pu in the marine environment.

The development of radioanalytical methods for low level radionuclides in marine environmental samples is presented. In particular, emphasis is placed on the introduction of extraction chromatography (EC) as a tool for improving the quality of results as well as reducing the analysis time. However, the advantageous application of EC often depends on the effective use of suitable preconcentration techniques, such as co-precipitation, to reduce the amount of matrix components which accompany the analytes of interest. On-going investigations in this field relevant to the determination of environmental levels of actinides, 137Cs and 90Sr are discussed.

Treatment with certain DNA-damaging agents induce a complex cellular response comprising pertubation of cell cycle progression and/or apoptosis on proliferating mammalian cells. Our studies were focused on the cellular effects of nickel (II) acetate, DNA-damaging agent, on Chinese hamster ovary (CHO) cells. Fragmented DNAs were examined by agarose gel electrophoresis and cell cycle was determined by DNA flow cytometry using propidium iodide fluorescence. Apparent DNA laddering was observed in cells treated with 240 microM nickel (II) and increased with a concentration-dependent manner. Treatment of nickel (II) acetate resulted in apoptosis which was accompanied by G2/M cell accumulation. Proportion of CHO cells in G2/M phase was also significantly increased in cells exposed to at least 480 microM nickel (II) from 57.7% of cells in the G0/G1 phase, 34.7% in the S phase, and 7.6% in the G2/M1 phase for 0 microM nickel (II), to 58.6%, 14.5%, and 26.9% for 640 microM nickel (II). These findings suggest that nickel (II) can modulate cellular response through some common effectors involving in both apoptotic and cell cycle regulatory pathways.

Instability of p53 in renal cell carcinoma (RCC) has not been previously associated with mutations, because 96% of clear-cell RCC samples in the database were present with the p53 wild type.1 However, inhibition of a major p53 regulator in humans doubles minute 2 homolog (HDM2) induced cell cycle arrest but not cell death in RCC.2 Therefore, overcoming p53 instability is a major issue in RCC treatment. Recently, our group3 and others4 have found that the expression level of transglutaminase 2 (TGase 2, E.C. 2.1.2.13) is generally elevated in most RCC cell lines, which deplete p53 into aggregates in the autophagosome, resulting in p53 depletion through autophagy.3 This p53 instability by TGase 2 regulation allows tumor cells to evade apoptosis and grow remarkably. TGase 2 is an enzyme that catalyzes an isopeptide bond between protein glutamine and lysine residues, resulting in a covalent cross-link.5 In normal physiological conditions, TGase 2 contributes toward regulating apoptosis from intruders or damages, including biological, chemical and physical challenges.6 However, TGase 2 knockout mice display impaired autophagy under starvation,7 although TGase 2 knockout mice present with an otherwise normal life.8, 9 Cancer cells adopt TGase 2 mediated autophagy for survival. TGase 2 competes with HDM2 for binding to p53; promotes autophagy-dependent p53 degradation in RCC cell lines under starvation; and binds to p53 and p62 simultaneously without ubiquitin-dependent recognition of p62. The bound complex does not have cross-linking activity. A binding assay using a series of deletion mutants of p62, p53 and TGase 2 revealed that the PB1 domain of p62 (residues 85–110) directly interacts with the β-barrel domains of TGase 2 (residues 592–687), whereas the HDM2-binding domain (transactivation domain, residues 15–26) of p53 interacts with the N-terminus of TGase 2 (residues 1–139).1 During translocation of p53 to the autophagosome through TGase 2 binding, cross-linking activity is not needed. This finding is in agreement with the observation that TGase 2 cross-linking activity occurs only in the autophagosome during autophagy.1, 7 This suggests that TGase 2 acts as a chaperone of p53 with a cross-linking catalytic activity. This interaction may result in rapid autophagy without consuming energy to tag ubiquitin on p53, as p62 is known to interact with ubiquitinated proteins (see Figure 1). This autophagy process is beneficial in which it supplies building blocks, including degraded p53, for cancer cells. Figure 1 (1) In normal cell, p53 stability depends more on HDM2 regulation due to no induction of TGase 2. (2) In cancer cell, TGase 2 is induced by stress such as hypoxia or starvation. The N-terminus of TGase 2 interacts with the N-terminus of p53 and, simultaneously, ... We recently reported that monotherapy using the TGase 2 inhibitor GK921 in a xenograft tumor model abrogated RCC growth through p53 stabilization.10 In addition to the increase in p53 stability due to TGase 2 inhibition, the administration of a DNA-damaging anti-cancer drug such as doxorubicin, remarkably induced apoptosis in RCC cell lines and sensitively reduced tumor volume in a xenograft model. Combination therapy with a TGase 2 inhibitor and a DNA-damaging agent may represent an effective therapeutic approach for treating RCC.

Strain DS4, a soil microorganism producing an oxygen radical-generating antibacterial substance, bruceanol B, was isolated, and cultural conditions for the antibiotic production were optimized. Bruceanol B was separated by ethyl acetate extraction, silica gel and Sephadex LH-20 gel column chromatographies, and high-performance liquid chromatography (HPLC). The highest antibacterial activity was obtained from medium containing 1.5% soluble starch and 2.0% yeast extract, cultured at 30°C for 3.5 days.